A Drop of Clarity: Liquid Biopsy-Mediated ctDNA Detection for Minimal Residual Disease and Beyond in Solid Tumors (Review)

Soheil Sardari,^1,*

1. IAUCTB

Introduction: As an alternative to tissue biopsy, liquid biopsy provides a non-invasive approach that enables detection of tumor cells or molecules originating from tumor tissues. Furthermore, this approach enables the detection and monitoring of minimal residual disease (MRD) in patients with cancer. MRD detection—mediated by liquid biopsy—has been playing a key role in clinical decision-making for hematological malignancies for decades, and recent efforts of research are aimed at the integration of MRD monitoring for solid malignancies into clinical decision-making. This could aid clinicians’ in assessing the need of every patient for adjuvant therapy after tumor resection, aiming to eliminate remaining disseminated tumor cells (DTCs), hence reducing chances of relapse. Liquid biopsy is capable of detecting traces of these cells in the peripheral blood of patients that current image-based assays deemed to be “relapse free”. Various biomarkers can be detected using liquid biopsy, such as circulating tumor cells (CTCs), circulating tumor DNA (ctDNA), and extracellular nucleic acids. In our review we focus on recent advances of liquid biopsy-mediated MRD detection with a focus on ctDNA as the biomarker for patients with solid tumors in clinical settings.

Methods: We used relevant keywords such as MRD, ctDNA detection, solid tumors, liquid biopsy, cancer prognosis, etc., on various clinical trial databases (most notably clinicaltrials.gov) and the PubMed database to identify relevant trials and studies mostly published 2016–2024 on subject matter with a focus on the most prevalent solid tumors.

Results: The ChemoNEAR study enrolled 76 patients with early-stage breast cancer (23 TNBC, 33 HER2+, 16 hormone receptor-positive, 4 unknown) and analyzed 598 longitudinal plasma samples using the NeXt MRD platform. Sample collection was executed pre- and post-surgery, after chemotherapy, and during follow-up to assess the correlation between ctDNA concentration and relapse risk in these patients. In this study, the ctDNA levels ranged from 204,900 to 2.2 ppm (median 296 ppm), with 40% of these detections being in the <100 ppm range (ultra-low). Subsequent to a 76-month follow-up (ranging from 5 to 113 months), MRD was detected in the 10 patients who relapsed, and the 58 patients with negative results for ctDNA did not suffer any relapse. The median lead time for ctDNA detection was reported to be 12.5 months (range is 1 to 60 months), showcasing the potential of utilizing ctDNA-based assays for adjuvant chemotherapy de-escalation. The randomized DYNAMIC Trial enrolled 455 patients with stage II (T3-4, N0M0) colon cancer with the aim of performing a tumor-informed ctDNA assay on the peripheral blood of each patient with the objective of assessing the utility of ctDNA-based assays in guiding adjuvant therapies for these patients. It was discovered that ctDNA-negative patients, despite not receiving chemotherapy, had no survival disadvantage (3-year recurrence-free survival rate of 92.5%) in comparison to ctDNA-positive patients who received adjuvant chemotherapy (3-year RFS 86.4%). These findings proposed that patients with stage II colon cancer can be spared adjuvant chemotherapy, highlighting the potential utility of ctDNA-based assays for guiding adjuvant therapies. In the TRACERx study, a median of 200 mutations were tracked in the resected neoplastic tissue of 197 NSCLC patients across the 1,069 plasma samples collected within 120 days of surgery. ctDNA was identified in 25% of patients, and 49% of relapse patients were ctDNA positive. Additionally, one instance of surveillance every 3-6 months revealed imminent relapse in 205 patients who had a history of negative ctDNA results. Due to low concentrations, ctDNA detection in prostate cancer patients has been challenging thus far. Additionally, prostate-specific antigen monitoring is a well-established procedure for these patients. Within this context, ctDNA detection must prove to be superior or complementary to PSA detection for relapse prognosis. The phase III multicenter randomized IMvigor010 trial enrolled patients with urothelial cancer with the aim to assess the efficacy of adjuvant atezolizumab versus observation. In ctDNA-positive patients (high relapse risk), atezolizumab was associated with better disease-free survival (DFS) and overall survival (OS) outcomes compared to the observation arm. Additionally, no difference in survival benefit was observed for ctDNA-negative patients in the observation arm compared to the treatment arm. Showcasing the value of ctDNA detection as a basis for adjuvant immunotherapy guidance in these patients.

Conclusion: ctDNA detection can predict MRD months before relapse, enhancing opportunities for early intervention and planning. Peripheral blood is not the only sample used in this procedure; the urine and cerebrospinal fluid can be analyzed for cell-free DNA fragments as a promising technique for urogenital tract and brain tumors, respectively. The inherent prognostic value of ctDNA detections lies in enabling clinicians to personalize treatment approaches for patients and provide insight into tumor characteristics. However, despite recent advances in studies and trials, the key challenge halting the integration of these assays into clinical practice is the lack of standardized procedures and absence of quality assurance criteria. Overcoming these challenges not only ensures the reproducibility of results but also builds trust for clinicians and patients as a procedure for reliable MRD detection. Additionally, tumor-informed ctDNA assays create a platform for personalized screening of tumor relapse in post-treatment settings. Furthermore, one of the key future challenges to overcome would be biomarker validity assessment for MRD detection to improve reliability alongside the development of higher sensitivity assays to minimize false positives, ensuring clinical efficacy.

Keywords: MRD, ctDNA, solid tumors, liquid biopsy, cancer prognosis

A Prevalence Of Scalp Dermatophyte Infections In Active Wrestling Boys (Review)

Samira zolfaghari kheirabadi,^1,*

1. Department of Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

Introduction: Introduction: Wrestling, one of the oldest sports, is popular among young athletes. Cutaneous infections are common in wrestlers, often due to increased exposure to pathogens and mechanical factors. The most significant infections include Tinea Gladiatorum caused by Trichophyton Tonsurans. This review focuses on the prevalence and effects of scalp dermatophyte infections, known as tinea capitis, in wrestling boys, discussing transmission, prevention, and treatment.

Methods: Materials and methods: This Study is A Review Were Wrestlers Of Wrestling Clubs. To Present An Overview Of The Existing Evidence Regarding The Occurrence Of Tinea In Wrestlers And Dermatophyte Contamination In Wrestling Halls. Databases Including Pubmed ,Google Scholar Were Searched From 2018 To 2024. Studies Focusing On Dermatophytosis Among Wrestlers And On The Presence Of Dermatophytes In Wresler Wre Included. Information From 10 Studies Has Been Collected And Analyzed. The flow of information through the different phases in this systematic review was depicted using a PRISM flow diagram.

Results: In this study analyzing 10 articles, 111 subjects had suspicious fungal skin lesions during physical examinations. The prevalence of such lesions among wrestlers was 34. 2%. Cutaneous infections pose significant risks for serious complications in combat athletes and lead to considerable time lost from practice and competition. The most reported infections in wrestlers are dermatophyte infections, specifically tinea gladiatorum, with trichophyton tonsurans as the main cause (92%). These infections can look different in wrestlers compared to the general community.

Conclusion: Athletes, especially in high-contact sports like wrestling, are at risk for skin conditions. T. tonsurans infections are spreading among combat sports clubs in Iran, highlighting the need for better prevention of tinea corporis gladiatorum. While tinea capitis mainly affects children, more cases in adults are seen. Physicians must know common skin disorders to ensure quick treatment and prevent spread. Athletic activity increases risk for fungal infections.

Keywords: Keywords: Dermatosis; Sports; Fungal Infection; Wrestling;

Advantages of Using CAR-MSCs in Cancer Immunotherapy (Review)

Batol Abbasi,^1,*

1. Tabriz University of Medical Sciences

Introduction: For many years, cancer treatment methods are usually chemotherapy, surgery, and radiation therapy. Despite these methods helping to cure the disease, most diseases have a poor prognosis. In recent years, there have been new advances in cell therapy with the production of chimeric antigen receptors (CARs) that destroy cancer cells. CARs are recombinant receptors that have been engineered so that immune cells can target and destroy specific antigens on the surface of cancer cells. Today, CAR-T cells (CAR-T), CAR-Macrophage (CAR-M), and CAR-NK (CAR-NK) are of great importance in cellular immunotherapy. However, there are several challenges in using CAR-related products. Recent studies have shown that mesenchymal stem cells (MSCs) can overcome cellular immunotherapy's disadvantages and improve CAR-related products' anticancer activity. Therefore, engineered MSCs (CAR-MSCs) can be used as a biological vehicle to improve the efficacy of CAR immune cells.

Methods: The main goal of CAR therapy is to direct cytotoxic lymphocytes against antigens expressed in tumor cells, which ultimately leads to tumor elimination. To enable immune cells to recognize cancerous cells bearing new antigens, the cells are engineered to express CARs that essentially link two functional domains: 1) extracellular antigen-recognition domain, and 2) intracellular stimulatory domain. The extracellular antigen recognition domain usually consists of a single-chain variable fragment (scFv) derived from a monoclonal antibody and, unlike true T-cell receptors (TCR), allows antigen recognition in an MHC-unrestricted manner (1). The extracellular domain is linked to an intracellular signaling domain, often the CD3-ζ chain of the TCR. Upon antigen recognition, the extracellular domain provides a stimulatory message to the intracellular domain, the intracellular ITAM domains are phosphorylated, and downstream signaling proteins are activated. Immune cells stimulated by CARs attack tumor cells and cause their lysis (2). Mesenchymal stem cells (MSCs), one of the most widespread cells in the human body, were first discovered in 1976 (3). In general, MSCs are known as pluripotent stem cells that can differentiate into adipocytes, chondrocytes, osteocytes, and other lineages (4). Also, they can adhere to plastic containers and multiply in laboratory conditions. These cells express CD73, CD90, and CD105 on their surface, but lack CD14, CD45, CD19, and HLA-DR. In addition to modulating the immune system, MSCs also repair damaged tissue. They migrate to the tissue through the chemokines that are released from the damaged tissue, and by differentiating into tissue-specific mature cells, they repair the damaged tissue (5). Here, we review the benefits of CAR and its applications in various cancers, as well as the unique ability of MSCs to enhance CAR immune cell activity.

Results: review

Conclusion: Today, CAR-immune cell therapies face several problems. These include problems such as dysfunction or cell exhaustion, immune effector cell-associated neurotoxicity syndrome, poor persistence of CAR-bearing cells, disease relapse, low efficacy against solid tumors, cytokine release syndrome, and immunosuppression. Researchers focus on MSCs due to their unique properties to discover novel approaches to overcome these challenges. MSC cells in The tumor microenvironment (TME) can affect tumor cells through direct cell-to-cell contact, MSC-derived exosomes and cytokines, or signaling pathways. Also, MSCs, MSC-derived membranes and MSC-derived exosomes can be used as carriers of chemotherapy drugs, oncolytic viruses, or therapeutic genes to precisely deliver cytotoxic agents to cancer sites. Thus, cytokines and chemokines secreted by MSCs make them an attractive adjuvant in CAR immunotherapy. CAR-MSCs also have a stronger killing ability that can improve the function of other modified cells, including CAR-T Cells, due to these unique characteristics.

Keywords: Chimeric antigen receptor (CAR), Mesenchymal stem cells (MSCs), Cancer, Immunotherapy

Antibacterial activity of hespertin against E.coli by in silico approach (Research Paper)

Zahra Najafdari,^1,*

1. Department of Microbiology, Faculty of Basic Sciences, Lahijan Branch, Islamic Azad University, Lahijan, Iran.

Introduction: Clinically manifested resistance of bacteria to antibiotics has emerged as a global threat to society and there is an urgent need to develop novel classes of antibacterial agents. DNA gyrase is an ATP-dependent enzyme and is very important in the pathogenic process of many microorganisms, such as E. coli. Negative super-helical twists can be formed into closed-circuit double-stranded DNA by the DNA gyrase. The inhibition of this group of proteins, which is known as DNA topoisomerases, is of interest as an antibacterial agent. Topoisomerase plays key roles in gene transcription, translation, recombination, and cell proliferation. Flavonoids are a group of secondary metabolites present in many plants. They are characterized by polyphenolic compounds consisting of two benzene rings joined by a three-carbon heterocyclic ring. This unique structure undergoes various modifications, enabling flavonoids to engage in many key biochemical interactions occurring in organisms. Flavonoids are well known to have various anti-inflammatory, antioxidant, antibacterial, and anticancer properties. The objective of this research is to use an in silico approach to determine the antibacterial activity of hespertin against E. Coli.

Methods: At the beginning of the study, the Discovery software was installed. The protein code: 1KZN for the target enzyme, DNA gyrase, was retrieved from the PDB website. The literature review showed that significant research has focused on this protein code. Protein structure in PDB format was opened in Discovery, and after removing unnecessary parts, was saved in the PDB format. The structure of hespertin, in PubChem with the code: 72281was selected for docking. The 3D structure on PubChem was downloaded in SDF format and opened in Discovery. However, this software did not support this format, so re-saved in PDB format. After preparing the compound, the docking procedure was done. For pharmacokinetic analysis, the SwissADME website was used. For analysis, the PDB file in Discovery was resaved in MOL format and then uploaded to SwissADME via the Choose File option. Running the analysis on SwissADME provided all the physicochemical and pharmacokinetic properties of the compound.

Results: After performing the docking procedure, it was observed that the hesperetin formed hydrogen bonds with various amino acids of DNA gyrase, which play a crucial role in stabilizing the protein structure. Specifically, the ligand establishes hydrogen bonds with asparagine 46, alanine 47, aspartic acid 73, asparagine 124, methionine 91, and valine 71. In physicochemical and pharmacokinetic studies of the compound, hespertin with the formula of C₁₆H₁₄O₆ and a molecular weight of 302.28 g/mol, has a total polar surface area (TPSA) of 96.22 Ų and a molar refractivity of 78.06, which are useful in predicting its absorption and permeability behavior in the body. In terms of water solubility, based on the Log S (SILICOS-IT) model, the compound has a Log S value of -3.53, indicating a solubility of approximately 0.0884 mg/mL (8.84e-02 mg/mL) or. This value classifies the compound as soluble according to this model. The compound exhibits high gastrointestinal (GI) absorption, indicating that it can be efficiently absorbed through the digestive tract. However, it does not have blood-brain barrier (BBB) permeability, meaning it is unlikely to access the central nervous system easily.

Conclusion: The studied compound in the molecular docking, physicochemical, and pharmacokinetic investigation showed good properties Therefore, the efficacy of this compound can be analyzed for further studies in vitro and in vivo.

Keywords: Antibacterial activity, hespertin, E.coli, silico approach, SwissADME

Antibiotic resistance of Pseudomonas aeruginosa (Review)

Setayesh Bahramifar,^1,* Hoda Sabati,²

1. Bachelor`s student ,Microbiology group,Faculty of Advanced Science and Technology,TehranMedical Science,Islamic Azad University,Tehean,Iran
2. Biotechnology and Biological Science Research Center, Faculty of Science, Shahid Chamran University of Ahvaz, Iran

Introduction: Pseudomonas aeruginosa is an aerobic gram-negative bacterium and is ubiquitous in soil, decaying organic matter, vegetation, water, hospital environments, and wet storage tanks. This bacterium is not part of the body's microbiota, but it can be found in various parts of the body, such as mucous membranes, respiratory tract and gastrointestinal tract. This bacterium can cause disease in humans, especially in conditions of reduced host defense (neutropenia, chemotherapy, and burn wounds). Pseudomonas aeruginosa is inherently resistant to many antibiotics and can become resistant to other antibiotics through horizontal transfer of resistance genes and mutations. Infections caused by this bacterium manifest in various forms, such as respiratory infections in hospital settings, especially ventilator-associated pneumonia, systemic infections, gastrointestinal infections, bone and joint infections, and dermatitis. Exposure to beta-lactam antibiotics such as Penicillin induces the expression of the AmpC gene in Pseudomonas, and beta-lactamase enzymes hydrolyze the beta-lactam rings of these antibiotics and inactivate them. The increase in antibiotic-resistant strains of Pseudomonas aeruginosa, especially multidrug resistance, has created many problems in treatment.

Methods: By reviewing bacterial data and articles published in Google Scholar, PubMed, and Scopus, based on this information, we found that by performing antibiotic susceptibility testing, the best antibiotic can be found for the treatment of resistant Pseudomonas.

Results: The results showed that Pseudomonas is resistant to beta-lactamase antibiotics, and AmpC beta-lactamase activity was evaluated as an important factor in the development of antibiotic resistance. For effectiveness, it is better to use a combination of antimicrobial drugs for treatment. For example, the antibiotics ciprofloxacin and ceftolozane tazobactam had a good synergistic combination.

Conclusion: The high prevalence of beta-lactam-resistant strains of Pseudomonas aeruginosa requires greater care in prescribing drugs and treating infections associated with this bacterium, because the indiscriminate use of broad-spectrum antibiotics destroys the body's natural microbial flora and causes overgrowth of resistant Pseudomonas.

Keywords: Pseudomonas aeruginosa, antibiotic resistance, beta-lactamase, AmpC, Infection

Antineoplastic and immunomodulatory mechanisms by parasites and their metabolites (Review)

Mohadeseh Ramzi,^1,* Fatemeh Ramzi,² Masoud Lahouty,³

1. Department of Medical Parasitolog and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
2. Department of Microbiology and Virology, School of Medicine, Urmia University of Medical Sciences, Urmia, Iran.
3. Pediatric Health Research Center, Tabriz University of Medical Sciences, Tabriz iran.

Introduction: Cancer is a life-threatening disease and the second leading cause of premature death worldwide. Annual reports indicate millions of new cases of cancer and mortality. Today, recent research and advances in cancer treatment and prevention methods offer hope in the fight against this disease. New studies indicate that a wide range of pathogens and their products in the framework of immunotherapy can be effective in stimulating immune responses, fighting cancer, and managing it. The aim of this study is to investigate the strategies of immune system-modulating power and anti-neoplastic activity of parasites and their metabolite products.

Methods: In this review, we searched databases such as Google Scholar, PubMed, and Science Direct from 2014 to 2024 for the terms parasite, worm, protozoan, parasite metabolites, anticancer, antitumor, and immunotherapy. Ultimately, we selected and thoroughly studied 18 articles.

Results: Although parasites are harmful organisms that can cause disease and even cancer in their host, they also have the power to modulate the immune system and have anti-neoplastic activity. Numerous studies on the anticancer mechanisms of parasites have demonstrated their reliance on multiple mechanisms and the release of their own side products. The most important of these actions is that they keep and target the immune system to fight infections, boost the number of CD8+ T cells and Tregs, help cancer cells die, and stop them from spreading and making new blood vessels. Parasites also employ molecular mimicry, and the sharing of antigenic epitopes among cancer cells results in anti-neoplastic activity. Researchers have also discovered that anti-parasitic antibodies can react with and destroy cancer cells. Therefore, they are considered potential candidates for the development of vaccines to combat cancer.

Conclusion: The successful and promising strategy of using parasites and their products to stimulate anti-tumor responses for the treatment of incurable cancers still faces numerous challenges. RRecently, researchers have proposed immunotherapy as a highly effective treatment for variousHowever, in order to progress these novel treatment approaches, we require extensive research and clinical trials to comprehend the interplay between parasites and cancer cells, their impact on healthy body tissues, and the creation of weakened parasite strains that target tumor tissues specifically.ery necessary.

Keywords: Cancer, parasite, cancer immunotherapy, anticancer, parasitic immunotherapy

Application of CAR-Treg in the treatment of inflammatory and autoimmune diseases (Review)

Batol Abbasi,^1,*

1. Tabriz University of Medical Sciences

Introduction: The immune system has evolved to recognize and combat only foreign agents while remaining unresponsive to the body’s cells and tissues. This state is known as immune tolerance. Autoimmune disorders, including Systemic lupus erythematosus, Autoimmune polyendocrine syndromes, Immunodysregulation polyendocrinopathy enteropathy X-linked syndrome, and many other autoimmune diseases result from defects in immune tolerance. Regulatory T cells (Tregs) are a subset of T cells critical for maintaining tolerance and immune homeostasis. Therefore, deficiency in the number or function of these cells causes uncontrolled immune responses and tissue destruction and can ultimately lead to inflammatory disorders such as autoimmune diseases, graft rejection, or graft-versus-host disease. Today, engineered Treg cells (CAR-Treg) have been proposed to treat such diseases. Here, we summarize recent studies using CAR Tregs in modulating immune responses in autoimmune diseases, transplantation and gene therapy, and future clinical applications.

Methods: Regulatory T cells (Tregs) are a subpopulation of CD4+ T lymphocytes that play an important role in the induction of peripheral tolerance to self and foreign antigens and suppress the immune response (1–3). Tregs are divided into two categories based on the place of distinction. Induced Treg (iTreg) are cells produced in peripheral lymphatic organs as a result of the detection of a local antigen outside the thymus, whereas normal Treg (nTreg) form within the thymus upon maturation. Both cell populations express FOXP3+ CD4+ CD25+ markers, and their suppressive activity is essential for establishing and maintaining stable immune homeostasis (3,4). Tregs can promote tumor development and progression by hindering the effector immune response (5–7). Tregs use multiple mechanisms to suppress the immune system: 1) signaling inhibition of CD80 and CD86 costimulatory molecules on dendritic cells (DC) via cytotoxic T lymphocyte 4 antigen (CTLA4), 2) secretion of inhibitory cytokines, 3) interleukin‐2 (IL-2) utilization through high CD25 (IL‐2 receptor α‐chain) expression, 4) metabolic modulation of adenosine and tryptophan, 5) direct killing of effector T cells (8,9). This ability of Treg therapy to promote and reshape immune tolerance makes them an attractive candidate for a cell-based therapeutic approach to minimize autoimmunity and/or promote the development of transplant tolerance (10). At first, CAR-engineered T cells were used to treat malignancies. The main goal of CAR therapy is to direct cytotoxic lymphocytes against antigens expressed in tumor cells, which ultimately leads to tumor elimination. To enable immune cells to recognize cancerous cells bearing new antigens, the cells are engineered to express CARs that essentially link two functional domains:1) extracellular antigen-recognition domain, and 2) intracellular stimulatory domain. The extracellular antigen recognition domain usually consists of a single-chain variable fragment (scFv) derived from a monoclonal antibody and, unlike true T-cell receptors (TCR), allows antigen recognition in an MHC-unrestricted manner (11). The extracellular domain is linked to an intracellular signaling domain, often the CD3-ζ chain of the TCR. Upon antigen recognition, the extracellular domain provides a stimulatory message to the intracellular domain, the intracellular ITAM domains are phosphorylated, and downstream signaling proteins are activated. Immune cells stimulated by CARs attack tumor cells and cause their lysis (12). Today, research has been done on CAR for the treatment of inflammatory and autoimmune diseases, which we will review.

Results: Review

Conclusion: Since Treg cells cause immune homeostasis and prevent the intensification of the immune response, they can be a suitable option to reduce various types of auto-inflammatory and autoimmune responses. Treatment with Treg cells would be significantly more effective if Treg cells were engineered to express a chimeric antigen receptor (CAR). Recent studies have shown that CAR-Tregs are effective as cell therapy in autoimmune diseases, hematopoietic stem cell transplantation, and organ transplantation. Since Treg cells that can be isolated from peripheral blood are limited, they can be expanded in vitro and produce sufficient numbers of cells for clinical use. Preliminary studies have shown that cell therapy using polyclonal Tregs in autoimmune patients can immunosuppression. However, it has been shown that antigen-specific Tregs, including CAR-Tregs and TCR-Tregs, are superior to polyclonal Tregs in suppressing the immune system. In addition, CAR-Tregs have advantages over TCR-Tregs as they are not MHC-restricted and are less dependent on IL-2. However, considering the exhaustion of CAR-Tregs and their rapid decline in vivo can limit their effectiveness in suppression. Since CD28 is critical for maintaining Treg homeostasis and plays an important role in Treg proliferation, differentiation, and survival, several studies have shown that second-generation CAR-Treg with a CD28-stimulatory domain to expand Tregs can ameliorate Treg-cell exhaustion.

Keywords: Regulatory T cells (Tregs), Chimeric antigen receptor (CAR), CAR-Treg, Inflammatory, Autoimmune

Associations Between ARID5B Polymorphisms and Acute Lymphoblastic Leukemia: Insights Into Genetic Susceptibility and Risk Prediction (Review)

Moein Kohkalani,^1,* Seyyed Amin Seyyed Rezaei,² Sima Mansoori Derakhshan,³ Mahmoud Shekari Khaniani,⁴ Akbar Amir Firouzi,⁵ Maqsoud Mehri,⁶

1. Department of Medical Genetics, Tabriz University of Medical Sciences, Tabriz, Iran
2. Department of Medical Genetics, Tabriz University of Medical Sciences, Tabriz, Iran
3. Neurosciences Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
4. Department of Medical Genetics, Tabriz University of Medical Sciences, Tabriz, Iran
5. Department of Medical Genetics, Tabriz University of Medical Sciences, Tabriz, Iran
6. Department of Medical Genetics, Tabriz University of Medical Sciences, Tabriz, Iran

Introduction: Acute lymphoblastic leukemia (ALL) is the most prevalent pediatric malignancy, accounting for approximately 25% of cancer diagnoses in children worldwide. Despite advances in treatment that have significantly improved survival rates, the incidence, clinical presentation, and outcomes of ALL vary across populations, reflecting a complex interplay of genetic, environmental, and socioeconomic factors. Among the genetic contributors, single nucleotide polymorphisms (SNPs) in the AT-rich interactive domain 5B (ARID5B) gene have been consistently implicated in modulating susceptibility to ALL. ARID5B, a transcription factor involved in chromatin remodeling and gene regulation, plays a critical role in lymphoid development, making it a plausible candidate for influencing leukemogenesis. This review synthesizes current evidence on ARID5B polymorphisms and their associations with ALL risk. By consolidating findings from diverse studies, we aim to help our understanding of the genetic mechanisms underlying ALL and their implications for risk prediction and personalized treatment strategies.

Methods: This narrative review was conducted to provide a comprehensive overview of ARID5B polymorphisms and their association with acute lymphoblastic leukemia (ALL) susceptibility. Relevant studies were identified through extensive searches in PubMed, Scopus, and Web of Science, focusing on peer-reviewed articles and genome-wide association studies (GWAS) published up to December 2023. Search terms included combinations of ARID5B, polymorphisms, single nucleotide polymorphisms, acute lymphoblastic leukemia, and ALL susceptibility. Studies were selected based on their relevance to ARID5B genetic variants and their implications for ALL risk across different populations.

Results: The relationship between ARID5B polymorphisms and ALL susceptibility varies across populations. In China, the rs7089424 (GG and GT genotypes) and rs10994982 (AA genotype) polymorphisms are linked to an increased risk of B-cell ALL (B-ALL), particularly in hyperdiploid subtypes. In Egypt, the CC genotype in rs4948488 and the AG and AA genotypes in rs2893881 show significant associations with ALL; the CC and AA genotypes are linked to T-cell ALL (T-ALL), while the AG genotype correlates with B-ALL. Among Mexican children, the rs10821936 and rs7089424 polymorphisms are strongly associated with a higher risk of ALL. In the United States, SNPs such as rs10821936 and rs10994982 confer an increased risk of B-ALL, with racial disparities in allele frequencies influencing disease outcomes. In European cohorts, rs10821936 and rs7089424 are significant risk factors for childhood B-ALL, with risk alleles nearly doubling susceptibility. These findings underscore the consistent yet population-specific roles of ARID5B SNPs, including rs7089424, rs10994982, rs4948488, rs2893881, and rs10821936, in modulating ALL risk.

Conclusion: ARID5B polymorphisms are key determinants of ALL susceptibility, particularly rs7089424, rs10994982, rs4948488, rs2893881, and rs10821936. These findings highlight the importance of population-specific genetic studies in understanding disease risk and inform the development of strategies for early diagnosis and personalized treatment to improve patient outcomes globally.

Keywords: Acute Lymphoblastic Leukemia, ARID5B Polymorphisms

Bioinformatic Identification of overexpressed hub genes in colon cancer as prognostic biomarkers (Research Paper)

Maryam Tashakori,^1,*

1. Department of Biology, Science and Research Branch, The University of Tehran, Tehran, Iran

Introduction: The fourth most common cause of death globally and the most prevalent gastrointestinal cancer in Iran and other countries is colorectal cancer. Each year, approximately one million new cases of colorectal cancer are diagnosed worldwide, and nearly half a million of these cases result in death from the disease. It is estimated that the incidence of cancer will increase by about 45% in developed countries by 2025. However, despite advancements in cancer biology and the use of combined therapeutic strategies for cancer patients, not much change has been achieved in these statistics, and patients with colon cancer still have an unfavorable prognosis. It has led the researchers to identify as many genes as possible specifically expressed in colon cancer using bioinformatics studies to identify these genes in the first step and, in the next step, evaluate the impact of these changes on the pathways that cause malignancy. These hub genes can be used as diagnostic biomarkers for this disease. Biomarkers are essential to the disease's early prognosis or preventing its progression. The purpose of this study, in the initial step, is to use bioinformatics studies on colon cancer to identify genes with a significant increase in expression in colon cancer compared to healthy people and to confirm these changes using bioinformatics databases. Finally, the identification of effective biomarkers in the early diagnosis of this disease.

Methods: Bioinformatics analysis was used to screen differentially expressed genes (DEGs) between normal and colon cancerous tissues to find overexpressed genes used for prognostic biomarkers in this disease. Firstly, the microarrays related to colon cancer were obtained from the GEO database(www.ncbi.nlm.gov/geo) GEO database is a functional genomics data repository, storing microarray and sequencing data, which is accessible to the public, then we selected Gene expression profile microarray datasets(GSE44861), this dataset containing 56 primary tissues (tumor samples) and 55normal human samples (normal adjacent noncancerous)based on Affymetrix HT Human Genome U133A Array, DEGs were obtained by R language analysis And the genes obtained from this software using STRING online database(https://string-db.org/) for protein-protein interaction network, and then key genes were exported and screened by using Cytoscape software. CytoHubba software was used to find hub genes that were highly connected between normal and cancer tissues, finally for data verification of this analysis we used the GEPIA database

Results: Gene expression profiles for normal and colon-cancerous tissues were obtained from the GEO databases. After data filtering, and normalization, 176overexpressed genes in colon cancer were identified, then we used string (https://string-db.org)to analyze these obtained data, and the gene interaction network was drawn. In the next step, the obtained information was saved in TSV format and finally, the data was analyzed with the software Cytoscape. Based on my results, ten hub genes were detected including SSPP1, CD44, MYC, MMP1, PTGS2, MMP3, SERPINE1, PLAU, CXCL8, and TIMP1. This choice was based on the MCC method. To ensure these expression changes, we used the GEPIA database (http://gepia.cancer-pku.cn/), and the data's significance was evaluated using this database.

Conclusion: The bioinformatic analyses performed on the mentioned genes show the significance of the increased expression of the selected genes between normal and tumor samples. These genes can be used as diagnostic biomarkers of increased expression in colon cancer. It should be noted that this in silico analysis was carried out in a bioinformatic manner and data confirmation should be done clinically at the in vitro level to obtain more reliable results.

Keywords: Colon cancer, in silico analysis, GEO dataset, String database, Cytoscape

Circulating Tumor DNA: Bridging the Gap Between Cancer Genomics and Clinical Practice (Review)

Mehdi hashemi,^1,* Roya Sinaei,²

1. Department of Medical Genetics, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
2. Department of Genetics, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.

Introduction: Circulating tumor DNA (ctDNA) has emerged as a revolutionary biomarker in oncology, providing a non-invasive means to monitor tumor dynamics and genetic alterations. As cancer genomics advances, the integration of ctDNA analysis into clinical practice offers a promising avenue for enhancing patient care. This approach allows for real-time insights into tumor heterogeneity and treatment response, potentially transforming the landscape of cancer diagnosis, treatment selection, and monitoring. This paper explores the methodologies employed in ctDNA analysis, the results obtained from recent studies, and the implications for clinical practice.

Methods: The analysis of ctDNA typically involves several key methodologies, including blood sample collection, DNA extraction, and sequencing techniques. Blood samples are collected from patients, and ctDNA is isolated from the plasma using various extraction protocols. Next-generation sequencing (NGS) is commonly employed to analyze the ctDNA for specific mutations, copy number variations, and other genomic alterations. These techniques enable the identification of tumor-specific alterations that can inform treatment decisions. Additionally, digital droplet PCR (ddPCR) and other sensitive assays are utilized to quantify ctDNA levels and monitor changes over time, providing insights into tumor dynamics and treatment response.

Results: Recent studies have demonstrated the clinical utility of ctDNA in various cancer types. For instance, ctDNA analysis has been shown to correlate with tumor burden and treatment response in patients with metastatic breast cancer, non-small cell lung cancer, and colorectal cancer. In a landmark study, researchers found that changes in ctDNA levels after treatment initiation were predictive of patient outcomes, highlighting the potential of ctDNA as a biomarker for monitoring therapeutic efficacy. Furthermore, ctDNA has been utilized to detect minimal residual disease (MRD) post-surgery, offering a means to identify patients at high risk for recurrence and guiding adjuvant therapy decisions.

Conclusion: Despite its promise, the integration of ctDNA into clinical practice is not without challenges. One significant issue is the standardization of testing methodologies, as variability in sample collection, processing, and analysis can lead to inconsistent results. Additionally, the sensitivity and specificity of ctDNA assays can vary, raising concerns about false positives and negatives. To address these challenges, ongoing efforts are focused on developing standardized protocols and guidelines for ctDNA testing. Moreover, clinicians must be trained to interpret ctDNA results accurately, distinguishing between clinically relevant mutations and those that may not impact treatment decisions. The clinical implications of ctDNA analysis are profound. By providing a non-invasive method to assess tumor genetics, ctDNA can facilitate personalized treatment strategies. For example, the identification of actionable mutations through ctDNA analysis can guide targeted therapies, allowing for more effective treatment options tailored to individual patients. Additionally, ctDNA can serve as a valuable tool for monitoring treatment response and detecting disease progression, enabling timely interventions. As the field of cancer genomics continues to evolve, the integration of ctDNA into routine clinical practice has the potential to improve patient outcomes and enhance the overall quality of cancer care. In conclusion, circulating tumor DNA represents a significant advancement in the field of cancer genomics, bridging the gap between laboratory research and clinical practice. While challenges remain in standardization and interpretation, the potential benefits of ctDNA analysis in guiding treatment decisions and monitoring disease progression are substantial. Continued collaboration among researchers, clinicians, and regulatory bodies will be essential to realize the full potential of ctDNA in oncology. As the integration of ctDNA into clinical practice becomes more widespread, it is poised to transform the landscape of cancer management, leading to more personalized and effective treatment strategies.

Keywords: Circulating tumor DNA (ctDNA); biomarker; cancer;

Evaluation of IL-18 in serum of Hepatitis C patients as probable biomarker and prognostic factor (Research Paper)

Fatemeh Mirzaei,^1,* kurosh kalantar,²

1. shiraz university of medical science
2. shiraz university of medical science

Introduction: Hepatitis C virus (HCV) is a hepatotropic RNA virus that is closely associated with chronic liver inflammation, fibrosis, and hepatocellular carcinoma.Resolution of viremia post antiviral therapy does not lead to protective immunity and therefore reinfections can occur. Immune cell detection of HCV activates signaling pathways that produce cytokines and trigger the innate immune response against the virus, preventing HCV replication and spread. Among them Pro-inflammatory cytokines such as Interleukin (IL)-18 have been found to play a role in eliminating Hepatitis C virus (HCV) infection. Nevertheless, fluctuations in the production balance of pro- and anti-inflammatory cytokines during the immune response can trigger various types of liver damage. Consequently, it is important to investigate IL-18 serum levels in hepatitis patients and their relationship with HCV infection.

Methods: total of twenty-nine newly diagnosed HCV+ patients without any prior antiviral treatment, along with 17 healthy controls, participated in our study. Biochemical indicators of liver disease were assessed using biochemistry assay kits. Serum levels of IL-18 were assessed using the ELISA method prior to and following treatment with pan genotypic direct-acting antivirals (DAAs

Results: Our findings indicated a statistically significant variation in serum IL-18 levels in HCV+patients (692.261 ± 48.76) in contrast to healthy controls (520.00 ± 44.73) (P=0.021). Nonetheless, there was no notable difference in IL-18 serum levels between the treated group and the untreated patients (P=0.74). No notable correlations were found between the amount of IL-18 and levels of liver enzymes

Conclusion: In summary, our study's findings show that serum IL-18 levels in HCV patients prior to treatment were significantly higher than those in normal controls and experienced a nonsignificant decrease following treatment. We believe that IL-18 plays a crucial role in eliminating the virus during the early stage, and it could aid in the virus's persistence if serum levels become low.

Keywords: HCV, IL-18, inflammatory cytokine, viral infection, Hepatitis

Exploring the Role of miRNAs in Colorectal Cancer Diagnosis and Therapy (Review)

Seyed Mohammad Kasra Esfahani,¹ Behrooz Yahyaei,^2,* Kimia Sadat Esfahani,³

1. Department of Medical Sciences, Shahrood Branch, Islamic Azad University, Shahrood, Iran.
2. 1Department of Medical Sciences, Shahrood Branch, Islamic Azad University, Shahrood, Iran. 2Department of Medical Sciences, Biological Nanoparticles in Medicine Research Center, Shahrood Branch, Islamic Azad University, Shahrood, Iran
3. Department of Genetics, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.

Introduction: Colorectal cancer (CRC) is the third most common cancer worldwide and the second leading cause of cancer-related deaths, presenting a significant global health challenge. The development of CRC is increasingly understood to result from a complex interaction of environmental factors, lifestyle and genetic factors. The high mortality rate associated with CRC can largely be attributed to several key factors, including the lack of early diagnosis, the extensive spread of metastasis, and the tumor's resistance to chemotherapy and radiotherapy. These elements collectively contribute to the difficulty in effectively managing and treating CRC. Genome-wide association studies (GWAS) have identified over 150 genetic risk variants linked to CRC. It can be expressed that approximately 90% of these GWAS-identified variants are located within non-coding or intergenic regions, implying that their influence on CRC risk is likely mediated through regulatory effects on gene expression. Recent studies have identified various non-invasive biomarkers for CRC, including circulating cell-free or exosomal non-coding RNAs, tumor-associated proteins, polygenic risk scores, and cell-free DNA. Among these, circulating miRNAs, small non-coding RNAs (17–25 nucleotides), are key regulators of gene expression through mRNA translation repression or degradation. miRNAs influence processes such as development, apoptosis and cancer. Stable in bodily fluids like serum and saliva due to their association with exosomes or proteins, alterations in circulating miRNAs have been detected prior to tumor diagnosis by conventional methods. MiRNAs significantly influence patient survival by regulating key processes like cell proliferation, apoptosis and immune response. Acting as oncogenes (oncomiRs) or tumor suppressors, they impact cancer progression by targeting genes involved in tumor suppression, angiogenesis and metastasis. The aim of this review is to analyze the role of miRNAs in CRC.

Methods: A total of twenty relevant articles investigating the role of miRNAs in colorectal cancer diagnosis and therapy were identified through searches on PubMed, Google Scholar and Scopus databases using predefined keywords. These articles were then selected for review and analysis.

Results: Numerous studies have demonstrated that aberrant activation of the MAPK signaling pathway is closely linked to the development, invasion and metastasis of CRC especially miR-373 appears to influence the expression of key proteins through various mechanisms, thereby modulating the invasive and metastatic behavior of CRC. MiR-1470 may be involved in various signaling pathways, including PI3K-AKT, cAMP, and MAPK. The PI3K-AKT pathway is known to play a critical role in the development of CRC. Exosomal miR-1470 shows promise as a potential biomarker for CRC, representing a valuable opportunity for the development of non-invasive diagnostic tools. In 2024, a systematic review and meta-analysis by Xu et al. identified miR-23, miR-92, and miR-21 as having high diagnostic sensitivity (87%, 82%, and 79%, respectively) and accuracy (87%, 81%, and 79%). Additionally, miR-29, miR-23, and miR-20 demonstrated strong specificity, with values of 86%, 86%, and 84%. In 2023, KrishnaPriya et al. suggested that miR-26b-5p could serve as a clinically relevant tumor suppressor miRNA, inhibiting cell survival and reducing cell migration and invasion during the metastatic cascade of CRC. As a result, miR-26b-5p may hold potential for clinical application in limiting the metastasis of colon cancer. In 2023, Chen et al identified two putative susceptibility miRNAs, miR-192-3p and miR-1307-5p. These miRNAs were shown to play regulatory roles in both promoting and inhibiting the proliferation, migration and invasion of CRC cells, thereby supporting their potential involvement in CRC sensitivity.

Conclusion: To sum up, miRNAs are essential regulators in CRC, influencing critical processes such as cell growth, migration and invasion. Due to their stability in bodily fluids, they hold promise as non-invasive biomarkers for early diagnosis and prognosis. The potential of miRNAs as both diagnostic tools and therapeutic targets offers exciting opportunities for improving CRC management. Further research is required to fully understand their clinical applications and to establish their effectiveness in CRC detection and treatment.

Keywords: Colorectal cancer, Diagnosis, non-coding RNAs, miRNA, Therapy

Fecal microbiota transplantation: review and update (Review)

Reyhane Namazi Yousefi,^1,*

1.

Introduction: The human gastrointestinal tract (GI) is colonized by many different species, which help the intestine digestion and preventing other pathogen colonization. Many GI diseases are connected to the gut microbiota, including Clostridium difficile infection (CDI), inflammatory bowel diseases (IBD) and many other important gastrointestinal disorders. CDI is one of the most common health acquired infections in western hemisphere, according to CDC. Clostridium difficile, a fastidiously anaerobic gram positive bacillus is the causative agent of CDI. While hospitalized patients, especially those receiving antibiotics therapeutically are at increased risk, currently multiple studies have proven fecal microbiota transplantation (FMT) as a successful therapy for recurrent and refractory CDI.

Methods: The guidelines in the United States and European consensus conference, both suggest using a donor questionnaire to meet the exclusion and inclusion criteria. Furthermore, standard donor screening protocols should be set up to lower the risks of infection transmission from the donor to recipient, and a suitable donor ought to receive both blood and stool examinations within 4 weeks before donation. A spouse or close relative was historically considered as an ideal FMT donor. Fecal material from the spouse might minimize the risk of infection transmission because shared environmental risk factors. Fresh fecal material should be processed within 6 hours of donor production, and it can be stored at room temperature until further processing. Approximately 50 g of fecal material is mixed with 150 mL of sterile normal sodium chloride by blender. The mixture is filtered with a filter or gauze to clear away large particulate matter, which may obstruct the endoscope channel. Finally, the filtrate is infused into 60-mL syringes (generally 4e5 tubes) and infused to the recipient’s GI tract.

Results: Fecal microbiota–based therapies are effective therapy to prevent recurrent C. difficile in select patients. Conventional fecal microbiota transplant is an adjuvant treatment for select adults hospitalized with severe or fulminate CDI not responding to standard of care antibiotics. Fecal microbiota transplant cannot yet be recommended in other gastrointestinal conditions.

Conclusion: The gut microbiota is the only “organ” that we can share with others. With the continuous advancement of microbiome theory and technology, FMT has made increasing effectiveness in treating intestinal and extra intestinal diseases, even those that were previously refractory. Despite these positive advancements, certain unresolved issues persist, that should be discovered in future.

Keywords: clostridium difficile, fecal transplantation

Immune Modulation and the Role of Graphene Quantum Dots in Breast Cancer Therapy: Innovations in Targeted Treatment (Review)

Alireza karimi,^1,* Bita fazel,²

1. veterinary graduate, faculty of veterinary medicine, University of Ahvaz, Ahvaz, Iran
2. veterinary graduate, faculty of veterinary medicine, University of Tehran, Tehran, Iran

Introduction: In modern society, cancer is one of the most serious diseases with a high fetal number in the world; the latest statistics for 2022 show approximately 20 million cases and 9.7 million deaths. It is expected to rise to 35 million by the end of 2025. Especially breast cancer is the first source of morbidity and mortality in women. Investigations have highlighted some cancer treatment approaches such as the genetic manipulation of cancer and immune cells, immunotherapy, cell therapy, use of cancer-killing viruses, and si-RNA; however, all the above methods have faced practical and financial barriers. Recently, the field of Nanomedicine has drawn much attention due to its innovative approach to the treatment of cancer. Among such materials, graphene quantum dots (GQDs) can be underlined as Nano-sized particles originating from carbon that has been exfoliated. Its unique characteristics include an increased surface-to-volume ratio and photoluminescence, which allows detection during therapeutic applications. Additionally, their quantum exclusivity and biodegradability make them suitable for treatment, not to mention as vectors for delivering other pharmaceuticals. This work studies the impact of GQDs on breast cancer cells originating from females.

Methods: A thorough study on the therapeutic approach of using QGDs on cancer, specifically breast cancer, In this review article we searched Google Scholar, PubMed, and Scopus using keywords that are often linked to each other, such as "GQDs," "breast cancer," "drug delivery," and "therapeutics," From the different studies, we studies papers mainly about GQDs nanoparticles on breast cancer and customizing cancer treatment.

Results: GQDs have emerged as a promising approach in treating a wide range of cancers as they exhibit properties such as biocompatibility, photoluminescence, and the ability to be functionalized for targeted therapies. Studies have shown that GQDs induce selective MCF-7 cell lines. Traditional methods like chemotherapy or radiotherapy have the potential to cause severe systemic cytotoxicity. GQDs are also functionalized to target specific cell markers in cancerous cells. In the case of breast cancer, this would improve targeted drug delivery to patients through estrogen receptors or Human Epidermal Growth Factor Receptor 2 proteins.They also address cancer cell resistance by conjugating with drugs to allow combination therapies targeting multiple pathways, improving drug retention via endosomal escape, and generating reactive oxygen species (ROS) to disrupt cellular defenses. Additionally, GQDs' photoluminescence enables real-time monitoring of drug delivery, optimizing treatment protocols based on individual responses. Moreover, GQDs have the potential in synergistic applications of photodynamic and photothermal therapies to enhance treatment efficacy with reduced dosages of drugs. Their biodegradability and safety profile make GQDs the standard-bearer in innovative, patient-specific cancer treatments, although challenges in scalability and clinical translation remain to be addressed.

Conclusion: GQDs have, therefore, brought great expectations in breast cancer due to their ability to couple targeted therapy with real-time monitoring functions. Their vast number of unique properties, from selective cytotoxicity to the overcoming of drug resistance, make them very superior alternatives to traditional therapeutic approaches. Future achievements in GQD research should focus on improving efficient drug loading, designing multi-modal therapies like chemotherapy with photodynamic and photothermal approaches, and scaling up production processes to ensure clinical feasibility. As a result, extensive preclinical studies need to validate their safety and efficacy in vivo, besides the development of standardized protocols. With continued advancements, GQDs could significantly improve cancer treatment outcomes, reducing side effects and enabling personalized therapeutic approaches for different patient populations.

Keywords: Nanomedicine, GQDS, breast cancer, drug delivery, Cancer Treatment

Innovations and Challenges in Molecular Point-of-Care Testing for Hepatitis C in High-Risk Populations: A Comprehensive Review (Review)

Niloofar Niroomand Firoozabad,^1,*

1. Medical university of Mashhad/Iran

Introduction: Hepatitis C remains a significant public health challenge worldwide, particularly among high-risk populations such as people who inject drugs (PWID). Despite the availability of effective treatments, there are considerable barriers to the diagnosis and treatment of hepatitis C, especially in low- and middle-income countries and among high-risk populations. The use of molecular point-of-care (POC) testing and dried blood spot (DBS) sampling has emerged as effective methods for improving diagnosis and linkage to care. This paper examines the potential of these innovative methods to address these challenges and enhance care for hepatitis C, providing an overview of innovations, performance, advantages, and challenges associated with these technologies.

Methods: A comprehensive review of studies on the performance of POC molecular tests and DBS sampling for hepatitis C was conducted. The review includes an analysis of the accuracy, sensitivity, specificity, and acceptability of these technologies among different populations, particularly PWID. The research spans studies conducted across various countries and settings to provide a broad understanding of the applicability and effectiveness of these tools.

Results: Studies indicate that POC molecular tests and DBS samples exhibit high sensitivity and specificity, making them effective for diagnosing hepatitis C and linking patients to care in non-clinical settings. These technologies are feasible and well-accepted among high-risk populations, showing promising results in increasing diagnosis rates and treatment initiation. For instance, one study highlighted that POC tests integrated with nursing care and peer support significantly improved HCV treatment uptake among PWID. Another study demonstrated the high accuracy and rapid turnaround time of POC tests, which are critical in settings with limited access to traditional laboratory facilities.

Conclusion: Molecular POC testing and DBS sampling for hepatitis C hold significant potential for enhancing diagnosis and care in high-risk populations. Addressing challenges such as high costs, the need for user training, and sustainable funding will be crucial for wider adoption. The review highlights the importance of continued development and evaluation of these technologies to reduce the burden of hepatitis C and improve treatment outcomes. This paper emphasizes the critical role of molecular POC testing and DBS sampling in increasing access to care and reducing mortality rates associated with hepatitis C. Further efforts are needed to support the development and distribution of these tools to achieve global health goals.

Keywords: Point-of-Care Testing (POC), Hepatitis C Virus (HCV), High-Risk Populations, Molecular Diagnostics

Innovative Biomarkers for Multiple Sclerosis: Translating miRNAs into Clinical Tools for Disease Progression (Review)

Samin Rashtchi Jabbari,^1,* Elaheh Hassani,² Amir Hossein Kiani,³

1. Tabriz University Of Medical Sciences, Department Of Genetics
2. Tabriz University Of Medical Sciences, Department Of Genetics
3. Tabriz University Of Medical Sciences, Department Of Genetics

Introduction: Multiple sclerosis (MS) is a complex neurological disorder with diverse clinical manifestations and progression patterns. The variability in MS is influenced by genetic, environmental, and molecular factors, with epigenetic mechanisms such as miRNAs playing a crucial role. miRNAs, small non-coding RNA molecules, regulate gene expression post-transcriptionally and are pivotal in immune response, inflammation, and neural repair in MS. Identifying miRNA signatures in biological fluids could enable early detection and help differentiate MS subtypes, guiding therapeutic strategies. This study explores the diagnostic and prognostic potential of miRNAs in MS by analyzing their expression in patient samples and correlating these findings with clinical and radiological markers.

Methods: A comprehensive literature search was conducted using PubMed, Scopus, and Google Scholar, focusing on studies published between 2010 and 2023. The research concentrated on human studies, especially those examining miRNAs as biomarkers for MS diagnosis, progression, or treatment outcomes. Studies included both observational research and clinical trials.

Results: Recent studies have highlighted miRNAs as significant players in MS pathogenesis, offering insights into disease mechanisms and potential therapeutic targets. miRNAs influence immune and neural cells by regulating multiple target genes. Though predominantly intracellular, miRNAs are stable in biological fluids like plasma, serum, and cerebrospinal fluid (CSF), even under harsh conditions such as freeze-thaw cycles and pH fluctuations. This stability makes them promising biomarkers for MS. Early studies identified that higher levels of miR-25-3p and lower levels of miR-320b were associated with benign MS after 10 years. miR-25-3p correlated negatively with pro-inflammatory cytokines, suggesting its role in modulating inflammation and potentially contributing to a milder disease course. Conversely, lower levels of miR-320b may support this outcome, though its exact function in disease modulation needs further study. In patients with progressive MS, miR-155, miR-338, and miR-491 were upregulated, indicating their involvement in disease progression. These miRNAs influence neurosteroid synthesis, suggesting that disruptions in this system may contribute to MS pathology. Additionally, miR-92a-1** has been recognized as a biomarker that differentiates relapsing-remitting MS (RRMS) from secondary progressive MS (SPMS) and from healthy controls. Its expression is linked to disease progression, making it a valuable tool for monitoring MS. Other biomarkers such as miR-143-3p showed reduced levels in progressive MS, providing a clear distinction from other MS subtypes. Moreover, increased serum levels of miR-92a-3p and miR-486-5p correlated with higher white matter lesion volumes, further indicating their role in disease progression. The evidence demonstrates that miRNAs can serve as innovative biomarkers, offering diagnostic and prognostic value across different MS subtypes. Differential miRNA expression is associated with key disease mechanisms, including inflammation, neurosteroid disruption, and neural cell dysfunction. miRNAs such as miR-25-3p and miR-320b are linked to milder disease courses, while miR-155, miR-338, and miR-491 are associated with progressive MS. Additionally, biomarkers like miR-92a-1** and miR-143-3p are subtype-specific, which could be useful for distinguishing MS phenotypes and predicting outcomes.

Conclusion: miRNAs have considerable potential as biomarkers for diagnosing and managing MS due to their stable presence in biological fluids and their association with key disease mechanisms. However, challenges such as variability in findings, lack of standardization, and the need for further validation across diverse populations must be addressed. Despite these challenges, miRNAs hold promise for improving personalized care and therapeutic strategies for MS, offering non-invasive diagnostic tools and enabling more targeted treatments.

Keywords: miRNAs (MicroRNAs), Multiple Sclerosis, Disease Progression, Biomarkers

Investigate the distribution of ABO blood groups in Thrombosis (Review)

Seyed Mohammadreza Seyedi,¹ Mohammad Joudaki,² Mohammad Ali Jalali Far,^3,*

1. Student Research Committee, Ahvaz Jundishapur University of Medical Science, Ahvaz, Iran.
2. Student Research Committee, Ahvaz Jundishapur University of Medical Science, Ahvaz, Iran.
3. Thalassemia & Hemoglobinopathy Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Science, Ahvaz, Iran.

Introduction: Thrombosis is the formation of a blood clot within a blood vessel that can block blood flow through the circulatory system, with an annual incidence of 1 in 1000 adults. Thrombosis occurs for a variety of reasons, including platelet activation, platelet aggregation, endothelial dysfunction, genetic mutations, and inflammatory responses. Thrombosis contributes to vascular occlusion events in the microcirculation, leading to acute and chronic organ damage. The ABO blood group system influences several factors, such as Von Willebrand Factor (VWF) and factor VIII levels, that are associated with the risk of thrombosis. The aim of this systematic review was to investigate the distribution of ABO blood groups in thrombosis.

Methods: This study was conducted based on the PICO criteria and in line with the research objective and in accordance with the PRISMA checklist. This systematic review included a comprehensive search from 2015 to 2024 in the databases PubMed, Web of Science, Scopus, Medline, SID, and the Google Scholar search engine. The search used MESH keywords including “Thrombosis”, “ABO” and Boolean operators. Two independent researchers then screened the retrieved articles based on the inclusion criteria.

Results: A total of 428 articles were identified through the initial search. After screening the titles and abstracts of the articles, the number of articles was reduced to 59 articles, and finally, according to the inclusion and exclusion criteria and after reviewing the full text, 23 articles were included in this study. Most studies show a different distribution of ABO blood groups in thrombosis. The ABO blood group system is closely related to the level of VWF, which is a polymer secreted by endothelial cells and megakaryocytes. VWF acts as a protective carrier of coagulation factor VIII and mediates adhesion between platelets and subendothelial collagen as well as aggregation between platelets, which consequently increases thrombosis. The distribution of ABO blood groups in thrombosis is different, with the rate of thrombosis in blood groups A and AB being higher than other blood groups, and blood group O having the lowest rate of thrombosis among blood groups.

Conclusion: ABO blood group is an important factor in determining the risk of thrombosis, which is associated with thrombosis through various mechanisms and has a different distribution. The probability of thrombosis in non-O blood groups is higher compared to blood group O, and this increased risk of thrombosis is extremely important in patients with cancer, heart disease, lung disease, and Sickle Cell Disease. However, given the limitations of studies conducted in this field, more and more diverse research is recommended in this field.

Keywords: Thrombosis, ABO

Investigating the anti-cancer effect of flavonoid Wogonin in inhibiting histone deacetylase enzyme (Research Paper)

Behnoosh Omidvar,^1,*

1. Department of Clinical Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

Introduction: Introduction: The histone deacetylase protein family, which includes 18 types and is divided into three major classes, was first found in a calf thymus extract and plays a critical role in the deacetylation of histone N-terminal lysine residues as well as non-histone proteins. Histone deacetylases (HDACs) are involved in many biological processes: gene transcription, protein stability, DNA repair, and angiogenesis. HDACs effect on gene transcription and expression. Abnormal activity of HDACs in pathological conditions, caused by gene or external stimuli mutations, disturbs the acetylation-deacetylation balance, resulting in nucleotide structure condensation. Thus, tumor-suppressor genes are downregulated, ultimately contributing to the development of tumors. Therefore, they are considered promising targets against cancer. Although HDAC inhibition does not necessarily increase gene expression, the acetylation/deacetylation ratio was enhanced in different cancer cells. Action mechanisms of HDAC inhibitors mainly revolve around cell cycle arrest induction, differentiation, cell death, angiogenesis reduction, and immune response modulation. HDAC8 is highly expressed in different tissues and organs, particularly in the kidney, brain, prostate and pancreas. Being a unique member of HDACs, it affects the epigenetic gene silencing mechanism and cancer development. Thus, it can be a significant target for potential therapeutic intervention against malignancies. HDAC8, for example, enhances the spread of breast cancer cells by affecting AKT/GSK-3β/Snail signaling pathways in the body. In vitro studies have also shown that HDAC8 knockdown inhibits the growth of cancer cell lines like A549, HeLa, and HCT116. Wogonin, a mono-flavonoid obtained from Scutellariae radix (the dried root of Scutellaria baicalensis Georgi), has been shown to have antioxidant, anti-inflammatory, and anti-tumor effects. One study in 2011, for instance, has demonstrated the correlation between different flavones and apoptotic pathways, suggesting them as a therapeutic option to induce apoptosis in malignancies by altering CDK9 function and Mcl-1 expression. Other studies have also indicated wogonin anticancer activities in human leukemia, breast cancer, and cervical carcinoma. This study aims to investigate the anti-cancer effect of flavonoid Wogonin in inhibiting histone deacetylase enzyme.

Methods: Methods: Accelrys Discovery Studio® Visualizer 3.5.0.12158 (Copyright© 2005-12, Accelrys Software Inc.) was installed to visualize molecules. The PDB format of target protein human HDAC8 complexed with Trichostatin A (ID:1T64) was downloaded from Protein Data Bank (http://www.pdb.org), and it was pre-processed by deleting hetero-atoms, water, and every protein chain other than protein chain B. Then, the optimized PDB file was used for auto-docking. The ligand wogonin (CID: 5281703) was searched on PubChem (https://pubchem.ncbi.nlm.nih.gov/) and the 3D structure was downloaded as an SDF format, which was then changed to a PDB format with the software. AutoDock Vina was used for the docking analysis. After the auto-docking operation, the ligand-protein binding sites were observed in a 2D diagram to see the covalent and non-covalent interactions in detail. To assess the pharmacokinetic traits including, bioactivity and toxicity of wogonin, the file was uploaded to SwissADME.

Results: Results: Docking analyses have shown 18 interactions in ligand-protein complex, most of which were electrostatic interactions (TRP B:141, GLY B:140, LEU B:264, LEU B:262, LEU B:301, GLN B:263 VAL B:261, ALA B:158, ASN B:156, HIS B:142, TYR B:174). Moreover, van der Waals interactions were seen between the ligand and 6 amino acids such as VAL B:185, VAL B:159, GLY B:302, LEU B:299, PHE B:189, ILE B:162. Additionally, in the hydroxyl group positioned at C-7, van der Waals forces exist between hydrogen molecule and LEU B:262 and oxygen molecule and LEU B:301, GLN B:263. Covalent bonds are formed at the C-5 hydroxyl group where two Glycine amino acids (GLY B:140) are involved. The same phenomenon has also happened between the phenyl group and isoleucine (ILE B:162). Wogonin (C16H12O5) is a dihydroxy- and monomethoxy-flavone with 284.26 g/mol molecular weight. This compound is moderately soluble in water and highly absorbed in the gastrointestinal tract. Pharmacokinetics evaluations have predicted CYP1A2, CYP2C9, CYP2D6, CYP3A4 inhibition, no blood-brain barrier (BBB) permeation, non-substrate of the permeability glycoprotein (P-gp) and low skin permeation. Furthermore, Drug-likeness was positive in five different rule-based filters with an oral bioavailability score of 0.55. Synthetic accessibility (SA) 3.15 implies an easy synthesis.

Conclusion: Conclusion: Overall, Wogonin had formed interactions in the active site of HDAC8 enzyme; however, further in vitro investigation is needed to achieve a promising result.

Keywords: Wogonin, Histone deacetylase, inhibitor, HDAC8, Cancer

Investigating the effect of chrysin in inhibiting melasma caused by tyrosinase activity (Research Paper)

Nazanin Imany Zadeh,¹ Forough Mehranpour,^2,*

1. Department of Biology, Faculty of Sciences, Zand Higher Education Institute Shiraz, Fars, Iran
2. Department of Biology, Faculty of Sciences, Zand Higher Education Institute Shiraz, Fars, Iran

Introduction: A macule is a flat, distinct, discolored area of skin less than 1 centimeter (cm) wide. It doesn’t involve any change in the thickness or texture of the skin. Areas of discoloration that are larger than or equal to 1 cm are referred to as patches. Macules can be caused by various conditions that affect the appearance of your skin, resulting in areas of discoloration. Conditions that are likely to cause macules are: vitiligo, sun spots, age spots, and liver spots, post-inflammatory hyperpigmentation. Melanin synthesis is known to be controlled by the rate-limiting enzyme tyrosinase, the role of this enzyme as the principal determinant of skin pigmentation is unclear. Results from studies with human melanocyte cultures derived from different racial skin types reveal an excellent correlation between the melanin content of melanocyte cultures and the in situ activity of tyrosinase. Melanocytes derived from black skin have up to 10 times more tyrosinase activity and produce up to 10 times more melanin than melanocytes derived from white skin. Chrysin belongs to the group of natural polyphenols. It can be found, among others, in honey, propolis and fruits and has a wide range of biological activities, including the prevention of oxidative stress, inflammation, neurodegeneration, and carcinogenesis. As part of the human diet, chrysin is considered a promising compound to be used to prevent many diseases, including cancers, diabetes, and neurodegenerative diseases such as Alzheimer’s or Parkinson’s. This study aims to investigate the effect of chrysin in inhibiting melasma caused by tyrosinase activity.

Methods: Discovery Studio software was installed. The PDB website was accessed, and the enzyme code 2Y9X was searched. The protein structure was downloaded and subsequently opened in Discovery Studio, where the structure of the protein was displayed. Using the shortcut Ctrl+H, a dashboard was activated, allowing all chains to be removed except for the A chain, active sites, protein groups, and ligand groups. As a result, these four components were retained. The final protein structure was saved as pro.pdb. The LP file was opened in Discovery Studio, and the following steps were performed: The script function was selected, followed by Ligand Interaction and then Show Ligand Binding. This step displayed the amino acids surrounding the ligand that are likely to form van der Waals interactions. Finally, the Show 2D Diagram option was used to visualize the ligand’s two-dimensional structure.

Results: After performing the docking operation, 13 interactions were established between amino acids and the ligand Among these, 7 amino acids participated in Vander Waals interactions, and 6 amino acids participated in electrostatic interactions. Vander Waals interactions: LEU A:255, TRP A:293 , GLU A:98, TYR A:97 , PHE A:90 , PHE A:292 , LEU A:303.Electrostatic bonds:ARG A:95 , ASP A:300 ,HIS A:94, VAL A:299, PRO A:91, HIS A:296.

Conclusion: Based on the results and the bonds created between chrysin and tyrosinase, further study of the effect of this compound in inhibiting melasma may be helpful.

Keywords: Tyrosinase inhibitors, Melanin synthesis, Natural polyphenols, Molecular docking,

Investigating the overexpressed genes of oral squamous cell carcinoma (OSCC) compared to adjacent non-tumor epithelia (Research Paper)

Samira Ameri Golestan,^1,*

1. Department of genetics, Faculty of Biological sciences and technology, Shahid Ashrafi Esfahani University, Esfahan, Iran.

Introduction: Oral squamous cell carcinoma (OSCC) is the most common cancer of the head and neck region and one of the 10 most common cancers in humans. According to the World Health Organization (WHO), Southeast Asia and Europe are among the most significant regions with high prevalence and mortality rates of OSCC. This type of squamous cell carcinoma of the oral cavity accounts for more than 90% of oral tumors. OSCCs have a highly variable clinical course, and because it is often diagnosed only after reaching an advanced stage, high rates of mortality, along with local recurrence, systemic metastasis, or secondary tumors, are still reported. Various environmental and genetic factors can lead to the development of OSCC. Environmental factors such as tobacco, alcohol, poor oral hygiene, HPV infection, and genetic predisposition. These factors can cause changes in gene expression, facilitating tumor growth, metastasis, and resistance to treatment. In particular, HPV infection can induce changes in the p53 and RB genes, which play a role in cell cycle regulation. These alterations in gene expression contribute to the progression of OSCC. Genetic factors can also contribute to the development of OSCC. Seven genes have been identified as potential molecular markers for oral cancer by comparing gene expression profiles between OSCC and normal epithelium tissues. These genes include NPM, CDK1, NDRG1, HMGCR, EF1A, NAC, and CHES1. NPM, CDK1, and NDRG1 were overexpressed in cancer tissues, while CHES1 was under expressed. Additionally, HOXA genes, particularly HOXA1, are more abundant in OSCC than in healthy oral mucosa. Overexpression of HOXA1 in human epithelial cells promotes cell proliferation, while its downregulation in OSCC cells reduces growth, highlighting its important role in tumorigenesis. In individuals with oral squamous cell carcinoma (OSCC), significant improvement with surgical, radiotherapeutic, and chemotherapy methods has not been observed. In this study, we aim to investigate the overexpressed genes to propose a new therapeutic approach. This study aims to investigate the overexpressed genes of oral squamous cell carcinoma (OSCC) compared to adjacent non-tumor epithelia.

Methods: First, GSEs of Oral Squamous Cell Carcinoma were extracted from the NCBI GEO database. After evaluating the different GSEs, GSE37991 was chosen for analysis due to its [specific features, such as sample size, quality, or other reasons]. Data from GSE37991 were processed using R software to identify the overexpressed genes in this type of cancer. These genes were then analysed for protein-protein interactions through the string database. The protein-protein interaction network obtained from string was transferred to Cytoscape software for visualization. Finally, the network was analysed using the CytoHubba tool in Cytoscape and five key genes were identified.

Results: After analysing the gene network in Cytoscape for Oral Squamous Cell Carcinoma (OSCC), 76 overexpressed genes were identified. In this cancer, five key genes, including MMP1, MMP7, MMP9, MMP12, and PLAU, were found to play significant roles.

Conclusion: In various studies, changes in gene expression have been observed across different types of cancers, with some genes being overexpressed while others are under expressed. In this study, five key genes were identified as overexpressed in Oral Squamous Cell Carcinoma (OSCC). It is recommended that further in-depth research, including in vitro experiments and laboratory analyses, be conducted on these genes to better understand their role in the early treatment of this cancer.

Keywords: OSCC gene expression, Overexpressed genes, Oral cancer biomarkers, human head and neck cancer, oral

Investigation of the mechanism of Interferon-I in platelet destruction and reduction in Immune Thrombocytopenic Purpura (Review)

Seyed Mohammadreza Seyedi,¹ Mohammad Joudaki,² Mohammad Ali Jalali Far,^3,*

1. Student Research Committee, Ahvaz Jundishapur University of Medical Science, Ahvaz, Iran.
2. Student Research Committee, Ahvaz Jundishapur University of Medical Science, Ahvaz, Iran.
3. Thalassemia & Hemoglobinopathy Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Science, Ahvaz, Iran.

Introduction: Immune Thrombocytopenic Purpura (ITP) is an autoimmune disease with an estimated prevalence of 9.5 to 50 per 100,000. In ITP, platelet glycoprotein epitopes IIb/IIIa and Ib/IX are covered by antiplatelet autoantibodies and eventually removed by the Reticuloendothelial System (RES), which ultimately causes a decrease and disruption of platelet production. Interferons (IFNs) are glycoproteins that play an important role as the first line of defense in the immune system. IFNs are classified into three groups I, II, and III, with IFN-I itself including IFN-α and IFN-β, which can play a role in platelet destruction and reduction. Given the role of IFN-I in platelet destruction and reduction, the aim of this systematic study was to investigate the mechanism of IFN-I in platelet destruction and reduction in ITP.

Methods: This study was conducted based on the PICO criteria and in line with the research objective and in compliance with the PRISMA checklist. This systematic review included a comprehensive search from 2015 to 2024 in PubMed, Web of Science, Scopus, Medline, SID, and Google Scholar search engine. The search used MESH keywords including “Interferon-I”, “Immune Thrombocytopenic Purpura” and Boolean operators. Two independent researchers then screened the retrieved articles based on the inclusion criteria.

Results: A total of 327 articles were identified through the initial search. After screening the titles and abstracts of the articles, the number of articles was reduced to 65 articles, and finally, according to the inclusion and exclusion criteria and after reviewing the full text, 24 articles were included in this study. Studies indicate the role of IFN-I in platelet reduction in ITP. Apoptotic substances activate plasmacytoid dendritic cells (pDCs), which produce IFN-α, which binds to the IFN-α/β common receptor (IFNAR), and ultimately phosphorylates JAK1, TYK2, and STAT. Therefore, IFN-α causes the differentiation of T cell clones into Th0 or Th1 rather than Th2 cells, which leads to increased activity of natural killer (NK) cells and IL-2 and IFN-γ, which cause platelet destruction. IFN-α also directly suppresses megakaryopoiesis and reduces platelet count by inhibiting thrombopoietin-induced signaling.

Conclusion: In ITP, IFN levels, especially IFN-I, are elevated, which can lead to excessive activation of T and B cells and the production of autoantibodies that suppress the production and attack platelets, ultimately leading to platelet destruction and reduction. However, given the limitations of studies conducted in this field, more and more diverse research is recommended in this field.

Keywords: Interferon-I, Immune Thrombocytopenic Purpura.

Kinetoplastid Membrane Protein 11 (KMP-11): A Candidate for Vaccine Development and Serological Detection of Leishmaniasis (Research Paper)

Roohollah Fattahi,^1,*

1. Department of laboratory and clinical sciences, Faculty of veterinary sciences, Ilam University, Ilam, Iran

Introduction: KMP-11 protein has been highlighted as a potential target for vaccine development or serological detection tool due to its immunogenic properties. This study investigates the physicochemical and antigenic properties of the KMP-11(TPP44494.1) utilizing bioinformatics tools, aiming to evaluate its viability as a candidate for vaccine development and serological detection of Leishmania spp, across diverse host species.

Methods: The antigenicity, allergenicity, and toxicity of the KMP-11 were assessed using Vaxigen, AllerTop, and ToxinPred servers, respectively. The ProtParam server was employed to analyze the physicochemical characteristics of the protein. Additionally, linear B-cell and cytotoxic T lymphocyte (CTL) epitopes were predicted using the ABCpred webserver and the IEDB database, contributing to a comprehensive understanding of the protein's potential in immunological applications.

Results: The analysis demonstrated that the KMP-11 is antigenic, with a Vaxigen score of 0.5097, and is classified as non-allergenic and non-toxic. Physicochemical characterization revealed a molecular weight of 25090.30 Daltons and an estimated half-life of 30 hours in mammalian reticulocytes in vitro, over 20 hours in yeast in vivo, and more than 10 hours in Escherichia coli in vivo. Additionally, the aliphatic index of 39.28 suggests that the protein has a moderate level of thermostability, while the theoretical isoelectric point (pI) was determined to be 6.15. The GRAVY index of -1.248 suggests a polar nature with good water interaction, indicating high solubility. The protein comprises 40 negatively charged residues (aspartic acid and glutamic acid) and 35 positively charged residues (arginine and lysine), with a molecular formula of C1112H1686N304O333S14 and a total atom count of 3449. The QuerySol scaled solubility value was determined to be 0.687 indicates that the protein is predicted to have high solubility.

Conclusion: our findings identified multiple linear B-cell and cytotoxic T lymphocyte (CTL) epitopes within the KMP-11 protein, all exhibiting high antigenicity, thereby highlighting their potential for application in vaccine development and serological detection of Leishmania spp.

Keywords: Leishmania spp., KMP-11, epitopes

Longitudinal Analysis of Escherichia coli Prevalence and Antibiotic Resistance in Nosocomial Infections: A Seven-Year Study in Ilam Province, Iran (Research Paper)

Alireza Pourrahim,^1,* Ali Hematian,² Khadijeh Najafi Ghobadi,³ Omid Raiesi,⁴

1. Student Research Committee, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran.
2. Department of Microbiology, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran.
3. Department of Biostatistics, Faculty of Health, Ilam University of Medical Sciences, Ilam, Iran.
4. Department of Parasitology, School of Allied Medical Sciences, Ilam University of Medical Sciences, Ilam, Iran.

Introduction: Escherichia coli is a leading cause of nosocomial infections, particularly urinary tract infections (UTIs) and bloodstream infections (BSIs). Its capacity to acquire resistance mechanisms, including extended-spectrum beta-lactamase (ESBL) and carbapenemase production, has led to increased morbidity and limited therapeutic options. High rates of resistance to 3rd and 4th generation cephalosporins, fluoroquinolones, and carbapenems pose severe challenges to patient outcomes and infection control strategies. This study aimed to comprehensively assess the prevalence of E. coli in hospital-acquired infections, the distribution across infection types, and trends in antibiotic resistance over seven years in Ilam Province, Iran. The findings provide critical insights for optimizing treatment protocols and strengthening infection control programs.

Methods: This descriptive-analytical study utilized data from the National Nosocomial Infection Surveillance System from March 2017 to March 2024 across 13 hospitals in Ilam Province. Nosocomial infections were identified based on national criteria, which included infections manifesting at least 48 to 72 hours post-admission in patients without evident infection symptoms or latent disease at admission. The dataset categorized infections into instrument-associated (e.g., VAP, CA-UTI, CA-BSI) and primary infection groups (e.g., UTI, BSI, PNEU). Antibiotic resistance patterns were analyzed for key antibiotics, focusing on resistance trends across years. Statistical analyses were performed using SPSS version 27 to identify prevalence, resistance rates, and temporal trends.

Results: E. coli contributed significantly to nosocomial infections, accounting for 346 cases (25.99%) in instrument-associated infections, including 105 cases of ventilator-associated pneumonia (VAP), 174 catheter-associated urinary tract infections (CA-UTI), 22 catheter-associated bloodstream infections (CA-BSI), and 45 other infections. In primary infection groups, E. coli was responsible for 672 cases (18.17%), with the majority occurring in UTIs (326 cases), followed by BSIs (77 cases), surgical site infections (SSI, 86 cases), ventilator-associated events (VAE, 144 cases), and pneumonia (PNEU, 25 cases). Antibiotic resistance analysis revealed critical challenges. Resistance to 3rd and 4th generation cephalosporins averaged 75.23%, with resistance rates consistently high, ranging from 66.66% (2018) to 83.33% (2017). Fluoroquinolone resistance averaged 63.54%, peaking at 88.23% in 2017 and gradually declining to 64.78% in 2023. Carbapenem resistance averaged 37.83%, with fluctuations from 18.18% (2021) to 52.94% (2020). These trends highlight the persistence of ESBL-producing strains and the increasing prevalence of carbapenem-resistant E. coli (CRE), underscoring an urgent need for effective interventions.

Conclusion: This study underscored the significant role of E. coli in nosocomial infections across various clinical settings in Ilam Province, particularly in CA-UTI and UTI cases. High and sustained resistance to cephalosporins and fluoroquinolones, along with the concerning upward trend in carbapenem resistance, reflects the growing burden of multidrug-resistant strains. The findings emphasize the necessity of robust antibiotic stewardship programs, enhanced infection control measures, and continuous resistance monitoring to mitigate the impact of E. coli-associated infections on patient health and healthcare systems.

Keywords: Escherichia coli, nosocomial infection, antibiotic resistance

MCHC discrepancy in Hereditary Spherocytosis (Review)

Seyedeh Sara Tabatabaei,¹ Mohammad Jafar Sharifi,^2,*

1. Department of Laboratory Sciences, School of Paramedical Sciences, Shiraz University of Medical Sciences
2. Department of Laboratory Sciences, School of Paramedical Sciences, Shiraz University of Medical Sciences

Introduction: Hereditary spherocytosis (HS) is a congenital hemolytic anemia, designated as a red blood cell (RBC) disorder where there are multiple inherited defects in membrane proteins alike ankyrin-1, α -spectrin, band 3 and protein 4.2. These mutations lead to morphology alternation, increased fragility of the RBCs and their destruction in spleen. Main hematological parameters used for HS diagnosis are hematocrit (HCT), reticulocyte count (Ret), hemoglobin (Hb), mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC). MCHC is computed indirectly by analyzers as Hemoglobin (g/dL) divided by Hematocrit (%) ×100. An average MCHC range reported to be 32 to 36 grams per deciliter (g/dL). While elevated MCHC is often associated with HS, discrepancies can arise in results obtained from some hematology analyzers, such as Mindray analyzer.

Methods: A literature search was conducted in PubMed and Google Scholar using the keywords "MCHC", "HS" and "Discrepancy".

Results: In patients with HS, the presence of spherocytes leads to an escalated MCHC level due to the reduction in surface area, eventually results in a higher concentration of hemoglobin per unit volume. Studies have illustrated the fact that the performance of hematology analyzers, including the Mindray system, can vary significantly based on their features of the blood sample. The Mindray analyzer might not consistently resonate MCHC increase because of its particular methodologies and characteristics. Mindray’s algorithms probably does not adequately account for this specific morphology of spherocytes, bringing about lower measures of hemoglobin or overestimation of hematocrit, thus reasons in falsely normal MCHC values despite the presence of the spheroidal RBCs. Moreover, the discrepancies in MCHC readings across different analyzers emphasize the demand for, particularly in cases where HS is suspected but MCHC values do not align with clinical expectations. In circumstances where MCHC and peripheral blood smear (PBS) has no mutual affair, additional diagnostic tests must be considered.

Conclusion: While elevated MCHC is a hallmark of HS, different analyzers may indicate diverse results for MCHC in HS patients. This discrepancy matters the most when HS is suspected but MCHC does not coordinate with the clinical manifestations and PBS, leading to misdiagnosis or delayed treatment. It is essential for laboratories to be aware of the limitations of their analyzers, and to adjust reference ranges based on their analyzed defects, furthermore confirm the suspected patients with additional tests like Osmotic fragility test (OFT) or Eosin-5′-maleimide (EMA); also, flowcytometry and DNA analysis are highly suggested to ensure the certainty of the diagnosis. This abstract highlight the importance of using multiple diagnostic approaches to confirm the presence of the condition, and mindful interpretation of the reports.

Keywords: Hereditary Spherocytosis, MCHC, Mindray analyzer, discrepancy

Metabolic dysregulation of CD4 T cell and Rheumatoid arthritis pathogenesis (Review)

Fatemeh Mirzaei,^1,* kurosh kalantar,²

1. shiraz university of medical science
2. shiraz university of medical science

Introduction: Rheumatoid arthritis (RA) is a chronic, systemic autoimmune disease characterized by inflammation of the synovial tissue and autoantibody production. In RA, CD4 T cells are the most abundant lymphocytes in the synovial tissue and play a central role in the inflammatory processes within the synovial membrane. Also, many metabolic alterations occur in RA T cells, contributing to the pathogenesis of the disease. In this article, we will review how metabolic dysregulation in RA CD4 T contribute the development of inflammation in the synovial tissue.

Methods: a systematic search was performed in various database including Scopus, PubMed and google scholar to find high quality original articles in this issue with different key word such as immunometabolism and Rheumatoid arthritis, T cell metabolism.

Results: One of the most notable metabolic changes in RA CD4 T cells is the dysregulation in glucose metabolism. The glycolysis pathway is impaired in RA T cells due to a defect in the induction of the PFKFB3 (6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3) enzyme. Therefore, compared to normal CD4 T cells, RA CD4 T cells have a lower rate of glycolysis and a reduced cellular concentration of ATP after activation. On the other hand, RA CD4 T cells increase the expression of glucose-6-phosphate dehydrogenase (G6PD), which is the first enzyme in the pentose phosphate pathway. Consequently, glucose is shunted from the glycolysis pathway to the pentose phosphate pathway, leading to the accumulation of building blocks and the reducing metabolite such as NADPH (nicotinamide adenine dinucleotide phosphate) in RA CD4 T cells. The increase of NADPH leads to the consumption of cellular reactive oxygen species (ROS), putting the cell in a state of reductive stress. Reductive stress in RA CD4 T cells hinders the proper activation of the cell cycle kinase ataxia telangiectasia mutated (ATM) and enables RA T cells to skip the G2/M cell cycle checkpoint, resulting in hyperproliferation. Moreover, ATM deficiency redirects naïve CD4 T cell differentiation toward TH1 and TH17 lineages, leading to the formation of an inflammation-prone T cell pool. Additionally, sustained activation of mTORC1(mechanistic target of rapamycin complex 1) occurred in RA CD4 T cells that commits cells to Th1 and Th17 differentiation. In addition to glycolysis, in RA CD4 T cells, oxidative phosphorylation is impaired due to a deficiency of the GDP-forming β-subunit of succinate-CoA ligase (SUCLG2), which is a mitochondrial enzyme that catalyzes the bidirectional transformation of succinyl-CoA into succinate and acetyl-CoA. Consequently, SUCLG2-deficient RA T cells accumulate acetyl-CoA, leading to tubulin hyperacetylation, which allows the production of hyperinflammatory and tissue-invasive T cells. Accordingly, in RA CD4 T cells, the NADPHhigh, acetyl-CoAhigh metabolic state and the higher expression of de novo fatty acid synthesis pathway enzymes, such as fatty acid synthase (FASN) and acetyl-CoA carboxylase (ACACA), trigger lipogenesis and lipid droplet deposition. As result, surplus lipids in RA CD4 T cells contribute to generation of invasive membrane ruffles and cellular uropods that promoting tissue invasiveness features.

Conclusion: There is several alterations in major metabolic pathways in CD4 T cells during RA pathogenesis. RA CD4 T cells shunt glucose from the glycolysis pathway to the pentose phosphate pathway that alter the concentration of different metabolite. Also, it was showed that RA CD4 T cells have metabolic features such as constant activation of mTORC1, enzymatic defect in mitochondrial and elevation in fatty acid synthesis. In conclusion, these metabolic reprogramming induce pro-inflammatory phenotype and hypermobility in RA CD4 T cells, enhancing their ability for invasion into the synovial microenvironment and progression of Rheumatoid arthritis pathogenesis.

Keywords: T cell metabolism, Immunometabolism, Rheumatoid arthritis, CD4 T cell

MicroRNAs in Breast Cancer: Their Role in Diagnosis, Prognosis, and Therapeutic Resistance (Review)

Seyyed Amin Seyyed Rezaei,^1,* Moein Kohkalani,² Mohammad Asgharzadeh,³ Vahid Asgharzadeh,⁴

1. Department of Medical Genetics, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
2. Department of Medical Genetics, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
3. Biotechnology Research Center and Faculty of Paramedicine, Tabriz University of Medical Sciences, Tabriz, Iran
4. Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran

Introduction: Breast cancer remains a major public health challenge, accounting for significant morbidity and mortality worldwide. Advances in molecular biology have identified microRNAs (miRNAs), a class of small non-coding RNAs, as key regulators of gene expression with critical roles in carcinogenesis, tumor progression, and therapeutic resistance. Their distinct expression patterns in breast cancer patients have positioned miRNAs as potential biomarkers for diagnosis, prognosis, and treatment monitoring. This review synthesizes current knowledge of miRNA roles in breast cancer, focusing on their potential as biomarkers and their clinical implications for diagnosis, prognosis, and therapeutic resistance.

Methods: This review analyzes 300 published articles from January 2017 to December 2023, of which 12 were selected based on relevance and quality. Articles were identified using the keywords miRNA, breast cancer, diagnosis, prognosis, and therapeutic resistance and evaluated to summarize current knowledge of miRNA expression patterns and their implications for breast cancer diagnosis, prognosis, and treatment.

Results: Dysregulation of miRNAs is a hallmark of breast cancer, with distinct patterns of upregulation and downregulation observed in patient samples. MiRNA-205, a tumor suppressor, is frequently downregulated in aggressive and metastatic breast cancer subtypes. Its reduced expression is associated with poor prognosis, higher tumor grade, and therapy resistance. Functionally, miRNA-205 inhibits epithelial-to-mesenchymal transition (EMT) by targeting key transcriptional repressors such as zinc finger E-box-binding homeobox 1 (ZEB1) and ZEB2, thereby reducing tumor invasiveness and metastatic potential. Additionally, it modulates pathways like PI3K/AKT and MAPK, suppressing proliferation and promoting apoptosis. Given its consistent downregulation, miRNA-205 serves as a promising diagnostic and prognostic biomarker. Preclinical studies show that restoring miRNA-205 expression using mimics or nanocarriers can re-establish its tumor-suppressive functions, underscoring its therapeutic potential. In addition to miR-205, miR-200c and miR-141, both implicated in breast cancer pathogenesis, show decreased expression levels in breast cancer patients compared to healthy individuals. Notably, miR-200c levels are elevated in stage 4 patients with low MIB-1 expression, while miR-141 expression is significantly increased in earlier stages (1–3), as well as in cases of lymph node metastases and HER2-negative tumors. These findings suggest that miR-141 and miR-200c may serve as prognostic indicators, with their altered expression profiles linked to disease progression and patient outcomes. Additionally, miR-10 b and miR-373 are markedly upregulated in the serum of patients with lymph node metastases, making them valuable biomarkers for assessing lymphatic involvement. In addition to their diagnostic and prognostic roles, miRNAs contribute to therapeutic resistance. For example, miR-29a and miR-222 have been shown to modulate breast cancer cell resistance to adriamycin and docetaxel, respectively. These findings underscore the multifaceted roles of miRNAs in breast cancer, spanning tumor biology and treatment response.

Conclusion: MicroRNAs hold significant promise as biomarkers for the diagnosis, prognosis, and therapeutic management of breast cancer. Their ability to predict and influence drug resistance further highlights their clinical relevance. By integrating miRNA profiling into breast cancer research and clinical practice, we can improve early detection, personalize treatment strategies, and enhance patient outcomes.

Keywords: Breast Cancer, microRNA, miRNA, prognosis

Modern and Noninvasive Methods for Diagnosis of Parasitic Infections in Children (Review)

Mohammad Taghi Ahady,^1,* Kimia Heydari Tajadod,² Seyyedeh Shaghayegh Abbaspour Haghighat,³ Hananeh Sarkhani,⁴ Mohadeseh Javadzadeh Damirchy,⁵

1. PhD of Parasitology, Department of Biology, Faculty of Basic Sciences, Ardabil Branch, Islamic Azad University, Ardabil, Iran
2. Pediatrician, Department of Pediatrics, Ardabil University of Medical Sciences, Ardabil, Iran
3. Department of Biology, Faculty of Basic Sciences, Ardabil Branch, Islamic Azad University, Ardabil, Iran
4. Department of Biology, Faculty of Basic Sciences, Ardabil Branch, Islamic Azad University, Ardabil, Iran
5. Department of Biology, Faculty of Basic Sciences, Ardabil Branch, Islamic Azad University, Ardabil, Iran

Introduction: Introduction: Detection of parasitic infections in children requires accurate and complication-free methods, and it is necessary to replace invasive diagnostic methods with non-invasive laboratory techniques. Modern and non-invasive diagnostic methods are necessary to prevent possible complications and risks of invasive methods and accurate and timely diagnosis of parasitic diseases. Using modern and non-invasive diagnostic methods is very important to reduce the suffering and stress of young patients and ensure early diagnosis and treatment. This study aims to investigate the latest non-invasive techniques for the diagnosis of parasitic infections in children.

Methods: Methods: In this systematic review, for collecting scientific information, PubMed, MagIran, Elsevier and Google Scholar data banks were referred to, totally 61 articles were collected, and 22 articles were selected for study due to the specialization of their content. The key words and phrases that used in the internet search were: Children's parasitic infections, modern and non-invasive methods, parasitic infections diagnosis.

Results: Results: The findings showed that immunochromatographic tests (ICT) are very suitable for rapid and on-site diagnosis of diseases such as malaria and giardiasis. As a non-invasive method, urine-based diagnostic assays can detect parasite antigens or antibodies in urine samples. These tests can be used to diagnose children with schistosomiasis. Giardiasis, amoebiasis, and enterobiasis are among the diseases that are often diagnosed using non-invasive stool tests. Methods such as enzyme-linked immunosorbent assay (ELISA) can improve the accuracy of these tests. Saliva-based assays are being developed for several parasitic diseases, including malaria. These tests are a painless alternative to blood tests because they are non-invasive and can cause fewer problems for children. Polymerase chain reaction (PCR) test is a very sensitive and specific diagnostic tool for diseases such as leishmaniasis and trypanosomiasis. Modern serological diagnostics can provide results quickly and with minimal complications, such as rapid diagnostic tests (RDTs) for malaria and toxoplasmosis.

Conclusion: Conclusion: Modern and non-invasive techniques offer significant advantages in terms of ease and convenience of implementation and accuracy in the diagnosis of parasitic infections in pediatric patients. Techniques such as PCR, improved imaging, urine and stool-based tests, saliva-based tests, and immunochromatographic testing have revolutionized accurate and rapid diagnoses. By using these techniques, it is possible to minimize the stress and fear and complications caused by performing invasive tests in children, and by early and accurate identification of parasitic infections, an effective and efficient treatment process can be ensured.

Keywords: Keywords: Modern and Noninvasive Methods; Children Parasitic Infections; Parasitic Infections Diagno

Molecular and mRNA-Based Cancer Vaccines: Pioneering Progress and Clinical Applications (Review)

Faeze Karimi,^1,*

1. Shiraz University of Medical Science, Department of Immunology

Introduction: Cancer is a major global health issue and the second leading cause of death in the U.S. Immunotherapy, which uses the immune system to fight cancer, includes treatments like immune checkpoint inhibitors, adoptive cell therapy (ACT), and cancer vaccines. Provenge, approved by the FDA in 2010, is the only approved tumor vaccine for castration-resistant prostate cancer. The four types of cancer vaccines are tumor or immune cell-based, peptide-based, viral vector-based, and nucleic acid-based. mRNA vaccines have also shown significant promise in cancer treatment. These vaccines include two main types which are non-amplifying and self-amplifying.

Methods: In this study, we review 23 of those research and review articles related to mRNA-based cancer vaccines in recent years and provide a general view of how effective these vaccines can be.

Results: mRNA vaccines face stability and efficiency challenges in in vivo delivery. So, we can use DOTAP/DP7-C liposomes that could function as a universal mRNA delivery system, offering a straightforward approach to enhance intracellular mRNA delivery and boost the immunostimulatory activity of dendritic cells. In vitro-transcribed mRNA tumor vaccines primarily target four types of molecules: (1) tumor-associated antigens like AFP and CA125, (2) tumor-specific antigens, and (3) tumor-associated viruses including HBV, HPV, EBV, and HCV. mRNA vaccines induce an anti-tumor immune response by directly stimulating tumor-specific T-cells and delivering tumor antigens to the immune system. They introduce a molecule that encodes viral proteins or neoantigens, which are translated into dendritic cells (DCs) and stimulate immune responses. Unlike conventional vaccines, they elicit strong interferon and CD8+ T-cell responses with the help of antigen-presenting cells in the MHC I pathway. mRNA vaccines can stimulate dendritic cell maturation by TLR signaling. In non-immune cells, RIG-I and MDA5 detect foreign mRNA, triggering cytokine and chemokine production and subsequently recruiting innate immune cells. They also modulate the tumor microenvironment to enhance the immune response. mRNA-based vaccines can also regulate immune checkpoint molecule expression in TME by tumor and immune cells. These vaccines also can promote MHC-I expression and balance M1 and M2 macrophages. Indeed, they reprogram M2 to M1 macrophages related more to inflammation and anti-tumor responses. Adjuvants in mRNA vaccines can be categorized into four types: 1) the inherent adjuvant properties of mRNA vaccines; 2) mRNAs that encode immunostimulatory molecules; 3) mRNAs that encode antibodies; and 4) adjuvants based on the components of the delivery carriers. Technological advancements allow mRNA to produce proteins that inhibit tumor growth and enhance immune responses. Restoring PTEN function in PTEN-null prostate cancer cells inhibits the phosphatidylinositol 3-kinase (PI3K)-AKT pathway and promotes apoptosis. It can be used as an mRNA-based vaccine which suppresses tumor cells. In clinical trials, the prostate cancer vaccines CV9103 and CV9104, which contain self-adjuvanted mRNA, were well tolerated and immunogenic. Further, some trials in the first phase recommend that mRNA-based vaccines could be well tolerated. Besides, research shows that RNA-LPX vaccination is an effective immunotherapy for melanoma patients with prior checkpoint inhibitor treatment. It highlights the potential of targeting non-mutant shared tumor antigens for broader cancer vaccines. mRNA-based vaccines coding cytokines are used in different types of cancers; like IFN-α. mRNA-based treatments could be developed for common skin conditions like non-melanoma skin cancer, leveraging the strong therapeutic effects of previous IFN-α protein therapies.

Conclusion: There are different types of immunotherapy for cancer; one is a mRNA-based vaccine. These vaccines enhance anti-tumor response with the help of APC. While therapeutic messenger RNA cancer vaccines are not yet approved for routine treatment, early clinical trials have shown promising results when these vaccines are used alone or with checkpoint inhibitors.

Keywords: mRNA-based vaccines, Cancer, Immunotherapy, treatment

Oral Biofilms (Review)

Alireza Farahnak,^1,*

1. Department of Biology, Science and Art, Yazd

Introduction: The oral cavity harbors hundreds of microbial species that are present either as planktonic cells or incorporated into biofilms. The majority of the oral microbes are commensal organisms. Those that are pathogenic microbes can result in oral infections, and at times can initiate systemic diseases. Biofilms that contain pathogens are challenging to control. Many conventional antimicrobials have proven to be ineffective. Recent advances in science and technology are providing new approaches for pathogen control and containment and methods to characterize biofilms.Although the use of medical devices has enhanced health care and improved the quality of life, there is, unfortunately, a myriad of diseases and infections that can be attributed to biofilms associated with medical devices. Microbes can colonize on a medical device surface and cause infections, and at times can even lead to malfunction of the device. Infections are caused by a wide variety of organisms. So far, Pseudomonas, Vibrio, Escherichia, Salmonella, Listeria, Streptococcus, and Mycobacteria have been identified as causing biofilm-induced infections

Methods: As the importance of studying biofilm growth becomes increasingly prevalent in medical research, the need for developing new high-throughput techniques for analysis and characterization is of utmost importance. Microwell Plate Assay One of the most common and effective methods for growing and quantifying biofilms in a high-throughput format makes use of 96-well microtiter plates. This method is reproducible, efficient, and cost-effective. The materials required for data acquisition, such as microtiter plates and plate readers, are standard equipment for most microbiology laboratories. Calgary Device The Calgary device is a variation of the 96-well format. The Calgary device is a method that allows for batch culture growth of biofilms and planktonic cells and measures the minimum inhibitory concentration (MIC), minimum bacterial concentration (MBC), and minimum biofilm eradication concentration (MBEC) of the cells. It is based on the use of a modified 96-well microtiter plate with pegs that are attached to a removable lid. The pegs are designed to be removable from the lid. This allows for microscopic observation of the biofilm structure. There are channels in the bottom chamber of the Calgary reaction vessel that allow for medium to flow across the pegs allowing for the formation of equivalent biofilms on each peg. Atomic Force Microscopy Another approach that has been used to study biofilm topography is atomic force microscopy (AFM). AFM uses an extremely sharp tip (probe) attached to a cantilever. A laser is reflected off the top of the cantilever’s flat surface and onto an optical lens that measures the movement of the cantilever and records a topographical image of the sample. There are two different ways to use AFM to measure surfaces: contact AFM and non-contact AFM. In contact AFM, the probe is dragged along the surface of the sample, and the bending of the cantilever is measured by the laser. In non-contact AFM, the probe oscillates just above the surface of the sample and as the tip approaches the surface, molecular attractions between the probe and the sample cause the oscillations to decrease. AFM has been used to characterize biofilm adhesion and binding strengths on various substrates.

Results: Emerging research is exploring novel strategies to control pathogenic biofilms, including the development of inhibitors that disrupt the biofilm matrix, targeting bacterial communication pathways (quorum sensing), and using specific antimicrobials that are more effective against biofilm-embedded bacteria. Maintaining a balanced oral microbiome is essential, as imbalances can not only lead to oral diseases but also have implications for systemic health, potentially influencing conditions like cardiovascular diseases and diabetes. In summary, oral biofilms are complex microbial communities that play a significant role in dental health. Understanding their development and implementing effective control measures are key to preventing associated diseases and promoting overall well-being.

Conclusion: Significant advances in our knowledge of biofilms have been achieved over the course of the past decade. The development of biofilms and their structure have been the subject of many studies. The relationship between microbial ecology and oral and systemic disease has been established. Continued efforts employing scientific and technological advancements will result in new diagnostic assays, preventative treatments, and therapeutic interventions.

Keywords: oral , biofilms , high-throughput analysis

Prebiotics, Probiotics, and Synbiotics: The Golden Keys to Gut Protection and Cancer Prevention! (Review)

Mohammad Mehdipour,^1,*

1. Mohammad Mehdipour1,* - Sepideh Mokabberi2 1 0009-0005-4794-2040/Department of Cellular& Molecular Biology,Comprehensive Health Resaerch Center, Islamic Azad University, Babol Branch, Babol, Iran

Introduction: Gastrointestinal (GI) cancers remain a significant global health challenge, despite advances in treatment methods. These cancers represent 25% of all cancers and account for approximately 9% of global cancer-related deaths. The human gut microbiota has been increasingly recognized as a key player in the development and progression of GI cancers, such as colorectal cancer (CRC), gastric cancer (GC), and esophageal cancer. As the understanding of the mechanisms behind tumorigenesis in the GI tract advances, the potential role of probiotics, prebiotics, and synbiotics in cancer prevention and treatment has gained attention. This review explores the impact of gut microbiota and the therapeutic potential of these biotherapeutics in modulating cancer progression, reducing chemotherapy-related side effects, and improving post-surgical recovery.

Methods: This review article has been prepared based on reviews and data analysis of articles from 2020 to 2023 in Nature and Science Direct magazines as well as Google Scholar and Pop Med databases. The search keyword are : Cancer,Gastric Cancer,Colon,Prebiotics,synbiotics,probiotics

Results: Research indicates that the gut microbiota plays a crucial role in GI cancer pathogenesis. Preclinical and clinical studies have shown that probiotics (especially Lactobacillus and Bifidobacterium species), prebiotics (such as inulin and oligofructose), and synbiotics (combination of probiotics and prebiotics) exhibit significant anti-cancer properties. These biotherapeutics can interfere with various tumorigenic mechanisms, such as inhibiting tumor cell proliferation, modulating the immune response, and altering metabolic pathways in the gut. Furthermore, the administration of probiotics and prebiotics in cancer patients has shown promising results in alleviating chemotherapy-induced toxicity and improving gut health. However, while animal and laboratory evidence is substantial, clinical evidence from human studies remains limited and inconclusive.

Conclusion: The evidence strongly supports the idea that gut microbiota influences the development of GI cancers and that probiotics, prebiotics, and synbiotics may offer effective strategies for cancer prevention and adjunctive treatment. Despite these promising results, further research, particularly well-designed randomized controlled trials with human participants, is necessary to solidify the clinical benefits of these biotherapeutics in cancer therapy. The integration of microbiome-targeted interventions into cancer treatment regimens could potentially reduce side effects, improve therapeutic outcomes, and enhance the overall quality of life for cancer patients.

Keywords: Cancer,Gastric Cancer,Colon,Prebiotics,synbiotics,probiotics

Prevailing of HPV-16 and 52 genotype in 2022–2023 in Sanandaj, Iran (Research Paper)

Leila Atefmehr,^1,* Mohammad Haddadi,²

1. Research Center for Clinical Virology, Tehran University of Medical Science, Tehran, Iran
2. Research Center for Clinical Virology, Tehran University of Medical Science, Tehran, Iran

Introduction: Introduction: Human papillomavirus (HPV) presents a potential threat to the onset of carcinogenesis in the cervix, anogenital regions, and oropharynx. HPV encompasses over 200 types, with at least 12 having the potential to cause cancer, impacting the majority of sexually active individuals. In this current research, we explore the occurrence and spread of HPV genotypes.

Methods: During this cross-sectional study conducted in Sanandaj, Iran from Feb 2022 to Aug 2023, diverse samples including oral, vaginal, and genital were collected from individuals referred to private laboratories in Sanandaj, Iran. After sample collection and DNA extraction (FAVORGEN, Taiwan), they were subjected to PCR and genotyping (MehrViru, Iran). The subsequent statistical analysis unveiled infection rates across diferent demographics and age groups. STATA (version 17) were used for statistical analysis. We examined infection rates across demographics using t-tests and Odds Ratio.

Results: Overall, 26% (249) out of 950 cases tested positive for HPV, with 69% of these classifed as high-risk. Among the examined population, 98% (933) were female, and 2% (17) were male. Females aged 31–40 exhibited the highest percentage of HPV prevalence (115/460) in the study with the majority of positive cases belonging to HR genotypes. The overall most frequent genotypes identifed were 6, 16, 52, 53, 51, 58, and 56. HPV-16 exhibited the highest frequency among HR genotypes, accounting for 42 (17%) occurrences, followed by HPV-52 with a frequency of 32 (13%).

Conclusion: Our fndings emphasize the signifcant prevalence of HPV among females, particularly in the 21–30 age group. The identifcation of high-risk genotypes, underscores the importance of targeted interventions for specifc age cohorts. The age-stratifed analysis highlights a consistent predominance of high-risk HPV across age groups, indicating the need for age-specifc preventive measures. These results contribute valuable information for designing efective screening and vaccination strategies, to alleviate the impact of diseases associated with HPV.

Keywords: HPV, Genotyping, Prevalence, Cancer, Cervical cancer.

Sperm Chromatin Dispersion (SCD): A Method for Evaluating DNA Damage in Infertile Men (Review)

Reza Valian,¹ Mohadese Farahani,^2,*

1. Urmia University
2. Arak University

Introduction: Male fertility is a crucial aspect of reproductive health influenced by various parameters including sperm count, motility, and morphology. However, sperm DNA quality plays a vital role in the success of both natural conception and assisted reproductive technologies (ART). Sperm DNA damage has been linked to reduced fertility, recurrent miscarriages, and fetal abnormalities. Traditional sperm analysis methods (such as counting and assessing motility) cannot provide accurate information about sperm DNA integrity. Given the role of oxidative stress, environmental factors, and systemic diseases in DNA damage, a need for precise techniques to evaluate this damage has emerged. Sperm Chromatin Dispersion (SCD) is an assay that measures sperm susceptibility to DNA denaturation. In essence, this test allows for the assessment of sperm DNA damage. It is a simple and rapid method for examining parameters related to sperm DNA quality. In this article, we review the significance and applications of this technique as well as the challenges that lie ahead.

Methods: A systematic search of Google Scholar was performed using pertinent keywords to identify articles published between 2015 and 2022. The retrieved articles were then categorized and subjected to a detailed analysis.

Results: Numerous studies have demonstrated that the Sperm Chromatin Dispersion (SCD) test serves as an effective diagnostic tool for assessing sperm DNA quality. The primary applications of this technique include: 1. Diagnosis of male infertility: Studies have shown that the SCD test aids in identifying men with sperm DNA damage. Damage to sperm DNA can result in fertilization failure, reduced blastocyst formation rates, and increased rates of recurrent miscarriage. 2. Improving outcomes of assisted reproductive technologies (ART): Evidence has indicated that using the SCD technique to select healthier sperm can increase the success rates of IVF and ICSI procedures. This technique is particularly valuable in patients who have experienced unsuccessful ART cycles. 3. Impact of environmental and occupational factors: Investigations have shown that this method can be used to assess the impact of environmental factors such as air pollution, occupational exposure to toxins, and smoking on sperm quality. 4. Monitoring treatments to improve sperm quality: Antioxidant-based therapies and lifestyle modifications have been associated with reduced sperm DNA damage. The SCD test, as a monitoring tool, evaluates changes in sperm quality after treatment. Despite its numerous advantages, the use of the SCD technique is also associated with some challenges: 1. Lower sensitivity and accuracy compared to other methods: The SCD test has lower sensitivity compared to more advanced methods such as TUNEL and SCSA. This may limit its ability to detect minor DNA damage. 2. Dependence on user experience: The results of this test are highly dependent on the skill and accuracy of the technician, which can lead to variability between different individuals. 3. Inability to determine the type of DNA damage: This method only shows overall DNA damage and cannot differentiate between single-stranded and double-stranded breaks. 4. Limited application in specific populations: In some studies, this test has yielded poor results for assessing DNA damage in populations with severe damage or abnormal sperm

Conclusion: The Sperm Chromatin Dispersion (SCD) test, as a simple, rapid, and cost-effective method for assessing sperm DNA damage, plays a crucial role in male infertility studies and improving the outcomes of assisted reproductive technologies. With broad applications in diagnosing DNA damage, evaluating the impact of environmental factors, and monitoring the effectiveness of treatments, this technique has become a popular tool in sperm analysis. However, limitations such as lower sensitivity compared to more advanced methods, dependence of results on technician experience, and inability to differentiate between types of DNA damage indicate that this method requires improvements in accuracy and reproducibility. Future research can reduce existing limitations and increase the reliability of this method by integrating the SCD method with new technologies such as fluorescent staining, utilizing automated analysis software, and standardizing kits. Overall, despite its challenges, the SCD test remains a key tool in male infertility research and clinics. With potential improvements, it has the potential for broader applications.

Keywords: Sperm Chromatin Dispersion DNA damage Infertility Sperm

Studying the antibacterial property of Sulfuretin compound with bioinformatic methods (Research Paper)

Seyedeh Zahra Seyedpour,^1,* Mohammad Kamali,² Fatemeh Shams Moattar,³

1. Department of Microbiology, Faculty of Basic Siences, Lahijan Branch Islamic Azad University, Lahijan, Guilan, Iran.
2. Department of Microbiology, Faculty of Basic Siences, Lahijan Branch Islamic Azad University, Lahijan, Guilan, Iran.
3. Department of Microbiology, Faculty of Basic Siences, Lahijan Branch Islamic Azad University, Lahijan, Guilan, Iran.

Introduction: DNA gyrase is from an enzyme of the topoisomerase category, which is related to the second sub-category and plays a role in intracellular processes. This enzyme is mostly found in prokaryotic cells, especially bacteria. The task of this enzyme is to fit the long DNA strand in the small space of the nucleus by compressing it. This enzyme is a good target for antibiotics because it has different functions in cells. DNA gyrase removes positive supercoils ahead of the replication fork. Therefore, it plays an important role in the replication and reproduction of microorganisms. Two important classes of antibiotics that act on this enzyme are (1) aminocoumarin and (2) fluoroquinolones. Aminocoumarin inhibit the enzyme's ability to bind ATP, which reduces the enzyme's activity. Fluoroquinolones prevent the joining of two DNA strands, thus leading to cell death. Flavonoids are a large family of polyphenolic plant compounds that occur naturally in fruits, vegetables, chocolate, and beverages such as tea and wine. There are six different types of flavonoids, each of which breaks down differently. Flavonoids participate in the elimination of toxins from our body. Sulfuretin is one of the main flavonoids produced by Rhus verniciflua. which was used as a traditional medicine in East Asia. Among the properties of Sulfuretin, it can be said that it improves brain function, has antioxidant properties, and also reduces allergic airway inflammation. Sulfuretin has medicinal effects such as anti-diabetes, anti-cancer, anti-inflammatory. Sulfuretin can be found in daffodil and cornopsis flowers.This study aims to investigate the antibacterial property of the sulfuretin compound using bioinformatics methods.

Methods: In this research, Discovery Studio software version 3.5 was used. For this purpose, the software was installed on a 5-core computer running the Windows operating system. Then, the three-dimensional structure of the desired protein with the code 1kzn was searched on https://www.rcsb.org and downloaded in pdb format. The downloaded file was analyzed using Discovery Studio software, and water molecules and non-protein atoms were removed from the protein. Our desired ligand, sulfuretin with the code 5281295, was searched on https://pubchem.ncbi.nlm.nih.gov and downloaded in SDF format, then converted to pdb format in Discovery Studio software. Using Autodoc software version 4.2, the ligand and protein were docked and saved in pdb format. The saved file was analyzed in Discovery Studio software and saved as a two-dimensional structure. Finally, the pharmacokinetic properties of the sulfuretin ligand were studied.

Results: Sulfuretin with the code 5281295, was uploaded to http://www.swissadme. Its molecular formula was C15H10O5, with a molecular weight of 270.24 g/mol and a TPSA (Topological Polar Surface Area) of 86.99 Ų. It had 5 hydrogen bond acceptors and 3 hydrogen bond donors. In terms of water solubility, it was soluble in water and its GI absorption was high. The interactions of amino acid residues in the ligand-protein complex were as follows: The amino acids PHE(A:169), VAL(A:43,120,123,167) LEU(A:130), ASN(A:46), ALA(A:47), and ILE(A:59,130) formed van der Waals bonds. The amino acids ASP(A:73), MET(A:91,166), GLN(A:72), THR(A:165), and VAL(A:71) formed electrostatic bonds, and only one amino acid, VAL(A:120), formed a covalent bond.

Conclusion: According to the obtained results, it is suggested to extract this compound with a high degree of purity and to be subjected to enzyme inhibition test for further investigations.

Keywords: DNA gyrase, Aminocoumarins, Fluoroquinolones, Flavonoids, Sulfuretin

Studying the antifungal effect of apigenin against the vital enzyme of Candida Albicans (Research Paper)

Niyayesh Nikkhooy,^1,*

1. Department of Microbiology, Faculty of Basic Sciences, Lahijan Branch, Islamic Azad University, Lahijan, Iran.

Introduction: One fungus that grows naturally in the body is Candida Albicans. This yeast lives in the mouth, skin, and intestines. Healthy bacteria in the body (microbiome) control the balance of Candida. When Candida is off-balance, the yeast overgrows and causes infection. Common types of infections caused by C. albicans include: thrush, vaginal yeast infection, and invasive candidiasis. One of the vital enzymes of Candida albicans in the pathogenesis is dihydrofolate reductase or DHFR. This enzyme reduces dihydrofolic acid to tetrahydrofolic acid by using NADPH as an electron donor. Tetrahydrofolate and its derivatives are essential for purine and thymidylate synthesis, which are important for cell proliferation and cell growth. Effective inhibition of DHFR produces a blockade in thymidine synthesis leading to “thymineless” death. One bioflavonoid that is mostly present in plants is apigenin. Apigenin is present principally as glycosylated in significant amounts in vegetables (parsley, celery, onions) fruits (oranges), herbs (chamomile, thyme, oregano, basil), and plant-based beverages (tea, beer, and wine). Apigenin has many interesting pharmacological activities and nutraceutical potential. Its properties as an antioxidant are well known, and it can also be a therapeutic agent to overcome diseases like inflammation, autoimmune, neurodegenerative disease, and even several types of cancers. In this research, we investigate the antifungal effect of apigenin against the vital enzyme of Candida Albicans.

Methods: In the first step, Discovery software was downloaded and installed. Dihydrofolate reductase protein (code 4hoe) was obtained from the PDB site. The protein was prepared for the AutoDoc process (water molecules and Hetatm were removed) and the file was saved in Protein Data Bank format on the Discovery program. Apigenin(code 5280443) was downloaded from the PubChem site and the file was saved in Protein Data Bank format on the Discovery program.The ligand file format was changed from Protein Data Bank to MOL and opened on the Swiss ADME site. The pharmacokinetic properties of the ligand were obtained from the Swiss ADME website (a table was prepared from them). Finally, the hydrogen bonds of the ligand-protein complex were analyzed and the color of the amino acid chains, hydrogen bonds, and ligand were changed.

Results: The ligand–protein complex had two chains (A, B) and a ligand group. There was a hydrogen bond between the amino acid asparagine 124 of the B chain and the oxygen atom of the ligand. Ligand pharmacokinetic properties included formula: C15H10O5, molecular weight: 270.24 g/mol, number of H-bond acceptors: 5, number of H-bond donors: 3, TPSA: 90.90 Ų, Log Po/w: (3.2P), class: soluble, gastrointestinal absorption: high.

Conclusion: According to bioinformatics studies, apigenin interacted with an amino acid asparagine 124 in the enzyme's active site and the ligand compound has good pharmacokinetic properties for drug designing.

Keywords: Apigenin, Dihydrofolate reductase, Hydrogen bonds, Ligand, Docking

Studying the inhibitory effects of LGIVL pentapeptide on BCL-2 using in silico analysis (Research Paper)

ZahraYazdani,¹ FatemehSholehvar,^2,*

1. Department of Biology, Faculty of Sciences, Zand Institute of Higher Education, Shiraz, Fars, Iran
2. Department of Biology, Faculty of Sciences, Zand Institute of Higher Education, Shiraz, Fars, Iran

Introduction: Introduction: Nowadays, various reasons for overcoming cancer cell apoptosis have been discovered. The BCL-2 family is the effective enzyme that causes cancer cells to escape from programmed death. These types of proteins are important indicators of human cancers.These enzymes are located on the mitochondrial membrane. It has been observed for the first time in 1980 in lymphoma cells. Variation in BCL-2 family genes in mice suggests that all mammalian cells are ready to commit suicide but are protected by one or more pro-survival family members to prevent this from happening. Anticancer peptides (ACPs), including natural and modified Peptides, have received a lot of attention in these years and appear as new therapeutic methods, and ACPs have also been considered for the production of drugs and vaccines. In addition to antimicrobial and antioxidant properties, oligopeptides have also shown good anticancer activities. Therefore, in this research, the inhibitory effects of LGIVL pentapeptide on BCL-2 were investigated using in silico analysis.

Methods: Methods: At first, to check the structure and type of protein bonds, Discovery software was installed on the system. In the following, by studying various articles, it was determined that the code related to protein BCL-2 is 4MAN. Using the pdb site, the structure of the protein was found and downloaded in a suitable format. Using discovery software, the protein structure was observed in pure form and the structure of ligands was observed. Then, water molecules and Hetatm were removed for a better analysis of the protein using the desired software. After further investigation, and docking process the protein-peptide complex, which is the complex of our desired ligand (pentapeptide) and protein, was obtained and the designed ligand (LGIVL) was docked. Finally, the hydrogen bonds formed between the ligand and the protein BCL-2 or even Pi interactions were checked. To photograph the new structure and make the hydrogen bonds clearer for better analysis, coloring, and necessary changes were done. Site Expasy was used to obtain the physicochemical properties of the desired pentapeptide (LGIVL).

Results: Results: According to the conducted research and the obtained results, the desired ligand was formed three hydrogen bonds with protein BCL-2. The first bond of protein TYR19 with VAL4 ligand and the second bond of protein LYS20 with LEU1 ligand and the last hydrogen bond of SER102 protein with LEU1 ligand were formed. As a result, three protein amino acids with the names of Tyrosine19, Lysine20, and Serine102 were observed in hydrogen bond formation. Also, the results of the physicochemical properties of this pentapeptide include Molecular weight: 513.68, Theoretical pI: 5.52, the estimated half-life: 5.5 hours (mammalian reticulocytes, in vitro), Instability index:-25.96, aliphatic index: 292.00, and Grand average of hydropathicity (GRAVY):3.180

Conclusion: Conclusion: The studied pentapeptide with the LGIVL sequence interacted with the BCL-2 protein and due to its unique physicochemical properties, it is proposed to carry out further studies under laboratory conditions to obtain more accurate and complete results for its use.

Keywords: Keywords: Cancer, Enzyme, Insilico analysis, Physicochemical properties, BCL-2, Pentapeptide

Studying the role and function of miRNAs as biomarkers and diagnosis during the process of infection with different species of Echinococcos (Review)

Fatemeh Ramzi,^1,* Massoud Lahouty,² Mohadeseh Ramzi,³ Zahra gahvechi amiri,⁴

1. Department of Parasitology and Mycology, School of Medicine, Urmia University of Medical Sciences, Urmia, Iran
2. Pediatric Health Research Center, Tabriz University of Medical Sciences, Tabriz Iran
3. Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
4. Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Introduction: Cystic Echinococcosis is considered one of the most dangerous zoonoses worldwide between humans and animals, which can cause serious damage to the host. Research conducted in recent years indicates that understanding the pathogenic mechanism and accurate diagnosis of Echinococcosis species still face many challenges. Therefore, today it is important to identify and use PCR-based techniques as well as miRNAs as potential and non-invasive biomarkers for the diagnosis, control and management of many parasitic diseases. Studies conducted on the miRNA sequences of different species of Echinococcosis have shown that these biomarkers have a potential role in biological processes including transcriptional and nucleotide regulation of gene expression, reduction of morphological complexity, and modulation of host immune responses during infection and response. They play a significant role in the stress caused by the host's immune system and also by targeting several genes and hormones in the host's body during infection, causing the rapid development and development of the survival of the parasite, especially in the larval stages in the host's body, and creating drug resistance. The purpose of this study is to investigate the role and function of miRNAs in the biological and pathological processes of the parasite in modulating the host's immune system during infection with different species of Echinococcosis.

Methods: For the review, the keywords of Cystic Echinococcosis, Echinococcosis granulosus, Echinococcosis multilocularis, inflammatory reactions, miRNA and host immune system modulating factors were used from Pubmed, Google Scholar databases from 2015 to 2024, and finally, articles, 20 The article was selected and studied.

Results: Studying the biology of different species of Echinococcosis parasites, how they grow and develop in the host's body, as well as their inhibition of the host's immune system responses and their interactions with the host, to identify their complex life cycle and their ability to adapt to different environments. Also, new approaches of molecular targets to check the resistance of anthelmintic drugs, develop vaccines, new biomarkers for accurate diagnosis at the species level, and foundation and management of new effective methods for treatment and disease control are very key. On the other hand, identifying the genes and miRNA sequences of Echinococcosis species in the larval stages of infection can help prevent irreparable pathological damage and reduce surgical cases and invasive treatment methods.

Conclusion: Studies conducted on different Echinococcosis parasite miRNAs during different stages of parasite development in the host's body have shown that these molecules effectively play an important role in differentiating parasite-offending genes, virulence, drug resistance and host-parasite interactions. Also, by regulating the expression of host genes and targeting them, it plays a key role in the development of inhibitory factors for the migration of macrophages, neutrophils, and dendritic cells, and by disrupting the regulation of their absorption and function, it reduces the lethal effect of the immune system on parasites. MiRNAs have specific functional roles in each stage of parasite development, which can even regulate the timing of parasite development. Therefore, the study and deeper understanding of the complex pathophysiology of Echinococcosis parasites at the molecular level and the investigation of new sequences of miRNAs can be used as a potential diagnostic biomarker for more accurate, simple and faster diagnosis of disease stages and investigation of the basic mechanisms of the initial development of the host body's defense cells. , finding new strategies developing appropriate treatment methods and investigating the emergence of drug resistance to help control the disease.

Keywords: Cystic Echinococcosis, Echinococcus granulosus, miRNA, immune system

The Application of Convolutional Neural Networks in Diagnosing Cutaneous Fungal Infections: A Systematic Review of Diagnostic Accuracy (Review)

Alireza Pourrahim,^1,* Omid Raiesi,² Abdollah Karimi,³ Mohammad Mahdi Pourrahim,⁴ Alireza Vasiee,⁵

1. Student Research Committee, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran.
2. Department of Parasitology, School of Allied Medical Sciences, Ilam University of Medical Sciences, Ilam, Iran
3. Department of Computer Engineering and Information Technology, Amirkabir University of Technology, Tehran, Iran.
4. Student Research Committee, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran.
5. Department of Nursing, Faculty of Nursing and Midwifery, Ilam University of Medical Sciences, Ilam, Iran.

Introduction: Cutaneous fungal infections are prevalent worldwide, posing diagnostic challenges in clinical and laboratory settings. These infections, including tinea infections, onychomycosis, and fungal keratitis, often rely on conventional methods like potassium hydroxide (KOH) examinations, microscopy, and culture, which can be time-consuming, resource-intensive, and subject to human error. Recent advancements in artificial intelligence, particularly convolutional neural networks (CNNs), have revolutionized the field of dermatology by providing automated, accurate, and real-time diagnostic tools. This systematic review explored the application, efficacy, and diagnostic performance of CNN-based models for identifying and classifying cutaneous fungal infections.

Methods: A comprehensive literature search was conducted across PubMed, Scopus, and Web of Science databases to identify studies applying CNNs for diagnosing cutaneous fungal infections using keywords such as "convolutional neural networks", "cutaneous fungal infections", "deep learning", "diagnosis", "artificial intelligence" and related MeSH terms. Inclusion criteria were studies published in peer-reviewed journals, focusing on CNN-based approaches for fungal skin, nail, or corneal diseases. Seven studies were included after assessing quality using the Quality Assessment of Diagnostic Accuracy Studies 2 (QUADAS-2) for diagnostic studies and the PROBAST tool for AI-related methodologies.

Results: The included studies demonstrated high diagnostic performance of CNNs in detecting cutaneous fungal infections. Reported accuracy ranged from 88.1% to 99.95% across various datasets, with sensitivity values between 70.2% and 99% and specificity ranging from 72.7% to 100%. Models applied to microscopic images achieved mean accuracy rates of 88.10% and 88.78%, outperforming dermatologists with an accuracy of 74.53% (p < 0.0001). Sensitivity and specificity for these models reached 75.04% and 93.78%, respectively. In the diagnosis of fungal keratitis, CNN frameworks achieved near-perfect accuracy, with values as high as 99.95%, while improving real-time diagnostic performance. For dermoscopic and clinical image-based diagnosis of onychomycosis, CNN models demonstrated sensitivity of 70.2% and specificity of 72.7%, achieving comparable accuracy to dermoscopic examination (AUC = 0.751). Overall, the included studies highlighted CNN models’ ability to provide superior accuracy, sensitivity, and specificity compared to conventional methods, including dermatological assessments.

Conclusion: CNN-based models have shown significant potential in accurately diagnosing cutaneous fungal infections, including tinea infections, onychomycosis, and fungal keratitis, using clinical, dermoscopic, and microscopic images. These models achieve superior diagnostic performance compared to conventional methods and, in many cases, outperform dermatologists in accuracy and specificity. Despite these promising results, large-scale, multicenter studies are required to validate CNN performance further and facilitate its clinical integration.

Keywords: Convolutional Neural Networks; Deep Learning; Cutaneous Fungal Infections; Artificial; Onychomycosis

The bioassay tests and mass spectrometry for evaluation of xenoestrogens in tuna fish (Research Paper)

nader akbari,¹ Shahrbano Rostami,² Mahmoud Ghazi-Khansari,³ Gholamreza Jahed-Khaniki,⁴ Nabi Shariatifar,⁵ Parisa Sadighara,^6,*

1. a Department of Environmental Health, Food Safety Division, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
2. Hematology-Oncology and Stem Cell Transplantation Research Center, Tehran University of Medical Sciences, Tehran, Iran
3.
4. a Department of Environmental Health, Food Safety Division, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
5. a Department of Environmental Health, Food Safety Division, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
6.

Introduction: Exposure to xenoestrogens through food is a significant concern. These compounds have estrogen-like activity and lead to cell proliferation. In this study, MCF-7 cells were utilized for bioassay tests and evaluation of cell proliferation of extracts prepared from canned tuna fish.

Methods: First, a pre-screening using the MTT test was done with the solid and liquid parts of the canned tuna fish. It was observed that the proliferation of cells in the solid part of canned tuna cans actually occurs in tuna meat. Therefore, this portion of canned tuna fish was selected for further studies. The binding affinity with ERα and ERβ receptors was determined through RT-qPCR, and the extract was analyzed via mass spectrometry to identify estrogenic components

Results: The study revealed an increased cell proliferation rate upon treatment with the canned tuna fish extract. 17β-estradiol was considered as positive control. The cell proliferation rate ranged from 27 % to 36.6 % compared to 17β-estradiol. Both ERα and ERβ receptors of MCF-7 cells were stimulated almost equally in the sample extract group. Mass spectrometry analysis identified bisphenol A (BPA) and other estrogenic compounds such as phytoestrogens, phthalates, polychlorinated biphenyls, natural and synthetic estrogen hormones

Conclusion: Our findings emphasize the necessity of using a combination of bioassay tests and analytical methods to carefully manage and evaluate the combined estrogenic effects of components in food samples

Keywords: Estrogenic activity E-screen Canned Fish Mass spectrometry RT-qPCR

The clinical characteristics, diagnosis, and treatment of Candida infections (Review)

Mahan Ebadpour,^1,*

1. Department of Microbiology, Faculty of Basic Siences, Lahijan Branch Islamic Azad University, Lahijan, Guilan, Iran

Introduction: Candida represents an extremely diverse group of fungal organisms. It reproduces asexually through a budding process. The texture of candida colonies varies by species, ranging from smooth and glistening to dry or wrinkled and dull. Candida yeasts are part of the normal flora in the human oral, intestinal, and vaginal mucosa, where they can colonize these areas. Yeasts of the Candida genus are linked to various clinical conditions, ranging from bloodstream infections (BSIs) and intra-abdominal candidiasis to deep-seated candidiasis and superficial infections. These microorganisms can also act as opportunistic pathogens. They detect suitable conditions to activate virulence factors and induce infection, commonly known as candidiasis. Candidiasis is a general term describing infections of the skin, oral cavity and esophagus, gastrointestinal tract, vagina, and vascular system of humans caused by fungi from the Candida genus. These infections can develop at any age mainly due to the rise of the AIDS epidemic, an increasingly aged population, higher numbers of immunocompromised patients, and the more widespread use of indwelling medical devices. C. albicans is the most pathogenic species and serves as the primary cause of candidosis. The pathogenicity of Candida is facilitated by various virulence factors, with key mechanisms including adherence to host surfaces (including medical devices), biofilm formation, and the secretion of hydrolytic enzymes such as proteases, phospholipases, and hemolysins. This review provides an overview of the Candida species involved, clinical aspects, diagnostic methods, and treatments available against candidiasis.

Methods: A comprehensive search was conducted in Google Scholar, Science Direct, Scopus, PubMed, and other databases to discover published articles related to the clinical characteristics, diagnosis, and treatment of Candida infections with search terms included, treatment of candida infections, clinical characteristics, and diagnosis of candida infections.

Results: Various approaches, with differing levels of success, have been employed to identify Candida species isolated from clinical samples. In addition to traditional morphological and biochemical methods, numerous advanced techniques such as molecular assays, MALDI-TOF mass spectrometry, antibody-based agglutination tests, and omics analyses have been effectively utilized for identifying Candida species and diagnosing clinical candidiasis. Combining these methods significantly enhances the accuracy of identifying Candida at both genus and species levels. Blood cultures can take several days to show positive results. Optimal outcomes for disseminated candidiasis are achieved when treatment starts within 12 to 24 hours of blood collection. However, cultures seldom yield positive results within this timeframe, rendering them too delayed and insufficiently sensitive to enable prompt and effective treatment.Beta-D-glucan (BDG) is a polysaccharide found in the cell wall of many fungi, including Candida. The BDG test detects the presence of this substance in the bloodstream, which indicates a fungal infection, particularly in immunocompromised patients. BDG is released into the bloodstream when fungal cell walls break down, making it a useful marker for systemic fungal infections. Polymerase Chain Reaction (PCR) is a molecular diagnostic method that amplifies specific DNA sequences to detect the presence of fungal pathogens, including Candida. PCR identifies the genetic material of the fungus, providing a precise and direct method of diagnosing fungal infections. Therapeutic options for invasive candidiasis are primarily restricted to four classes of systemic antifungals: polyenes, the antimetabolite 5-fluorocytosine, azoles, and echinocandins. Among these, azoles and echinocandins are particularly effective, well-tolerated, and therefore the most commonly used treatments of Candida infections. Azole antifungal agents inhibit the 14-α-demethylase enzyme, a critical mediator in converting lanosterol to ergosterol within the fungal cell wall. Polyenes, such as Amphotericin B bind directly to ergosterol in the fungal membrane, disrupting its integrity. Echinocandins, another class of antifungals, target the fungal cell wall by inhibiting the synthesis of 1,3-β-D-glucan (BDG), exerting fungicidal effects, particularly against Candida species. Flucytosine (5-fluorocytosine) acts by disrupting fungal DNA and protein synthesis, inhibiting their growth and replication.

Conclusion: Based on the conducted analysis and the reviewed literature, Candida is identified as a significant cause of various diseases. A comprehensive understanding of its pathogenic mechanisms and diagnostic methods is essential, especially for early detection, which ensures timely and effective treatment. Consequently, developing and implementing advanced and innovative diagnostic methods is imperative to enhance the precision and efficiency of Candida identification, underscoring its critical relevance in medical research and clinical practice.

Keywords: Fungi, Candida, Infections, Candidiasis, C. albicans, Diagnosis.

The Hidden Influence of Gut Microbiota on Outcomes in Hematologic Malignancies: A Journey of Discovery (Review)

Elmira Zarei,¹ Ayda Zarei,² Azade Omidkhoda,^3,*

1. Department of hematology and blood banking, School of Allied Medical Sciences, Tehran university of Medical Sciences, Tehran, Iran
2. Department of Laboratory Sciences, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran
3. Department of hematology and blood banking, School of Allied Medical Sciences, Tehran university of Medical Sciences, Tehran, Iran

Introduction: Hematologic malignancies (HMs) characterized by the uncontrolled growth of abnormal blood cells, posing significant clinical challenges due to their heterogeneity and variable treatment responses. In 2020, HMs contributed notably to the global cancer burden, with non-Hodgkin’s lymphomas at 2.8% and leukemias at 2.5% of global cancer incidence. Despite advancements in therapies improving survival rates—such as a one-year survival rate of 91.8% for chronic lymphocytic leukemia—treatment often leads to adverse effects, including gut microbiota dysbiosis due to chemotherapy, which can impact drug efficacy and toxicity. Researchers are exploring strategies to manipulate gut microbiota to enhance treatment outcomes, as changes in microbial diversity can predict relapse risks and infections. This review emphasizes the need for further investigation into microbiota modulation as a potential therapeutic approach for patients with hematologic malignancies.

Methods: Studies related to microbiota and hematological malignancies published from 2020 till 2024 was conducted using MEDLINE and SCOPUS.

Results: The review highlights the complex relationship between hematological treatments and the gut microbiota, revealing how therapies like chemotherapy, radiation, and hematopoietic stem cell transplantation (HSCT) disrupt microbial balance and cause adverse effects. Chemotherapy agents such as 5-fluorouracil and methotrexate damage the mucosa and alter gut microbiota, increasing pathogenic bacteria like Enterococcus and Escherichia while decreasing beneficial species like Faecalibacterium prausnitzii and Bifidobacterium spp. This dysbiosis can compromise immune function and exacerbate side effects like mucositis and diarrhea. Certain bacteria, including Lactobacillus and butyrate-producing species, may offer protective effects against chemotherapy damage. Additionally, beneficial bacteria from the Firmicutes and Bacteroidetes phyla influence drug metabolism and efficacy. Fecal microbiota transplantation (FMT) shows promise in restoring microbial diversity and mitigating complications. The review calls for more research to address study limitations and emphasizes the need for personalized therapeutic strategies that enhance treatment efficacy and minimize adverse outcomes in cancer care.

Conclusion: Research on hematologic microbiota highlights their potential as biomarkers and therapeutic targets in hematopoietic malignancies. Strategies like probiotics, prebiotics, and fecal microbiota transplantation may restore balance and enhance treatment efficacy, warranting further investigation.

Keywords: Hematologic Neoplasms, Microbiota, Probiotics

The Role of Biomarkers in the Diagnosis and Prediction of Prostate Cancer: A Review (Review)

Sayyid Ali Hosseini,^1,*

1. Faculty of medicine, Mashhad University of Medical Sciences, Mashhad, Iran

Introduction: Prostate cancer is one of the most common types of cancer in men, and early diagnosis can significantly improve treatment outcomes. Biomarkers have emerged as important tools in diagnosing, predicting, and monitoring disease progression. This study aims to review the role of biomarkers in the diagnosis and management of prostate cancer.

Methods: This study reviews articles published in the PubMed, Scopus, and Web of Science databases. Articles published from 2010 to 2024 that examined biomarkers related to prostate cancer were included. Information regarding the type of biomarker, diagnostic methods, sensitivity, and specificity was collected.

Results: Results show that Prostate-Specific Antigen (PSA) remained the most widely used biomarker for initial screening and diagnosis. The review highlighted that while PSA is effective, its specificity is limited, leading to false positives and unnecessary biopsies. The optimal PSA threshold for biopsy recommendation was found to be 4 ng/mL, although this varies based on age and other risk factors. PCA3 (Prostate Cancer Antigen 3) emerged as a promising urinary biomarker, beneficial for patients with prior negative biopsies. Studies indicated that PCA3 had a sensitivity of approximately 60-70% and specificity of around 70-80%, making it a valuable adjunct to PSA testing. The 4Kscore test, which combines four kallikrein markers, showed enhanced predictive capability for aggressive prostate cancer. This test demonstrated a sensitivity of 93% and a specificity of 41% for detecting high-grade disease, providing clinicians with additional information to guide biopsy decisions. Emerging biomarkers such as AR-V7 (Androgen Receptor Variant 7) have shown potential in predicting resistance to androgen receptor-targeted therapies. Studies indicated that patients with AR-V7-positive tumors had a significantly lower response rate to treatments like abiraterone and enzalutamide. MicroRNAs (miRNAs) also appeared as promising candidates in the biomarker landscape. Specific miRNA profiles were associated with prostate cancer progression and could potentially serve as non-invasive diagnostic tools. Moreover, the combination of multiple biomarkers was shown to enhance diagnostic accuracy significantly. For instance, using a panel that includes PSA, PCA3, and 4Kscore improved the area under the curve (AUC) for predicting prostate cancer compared to any single biomarker alone.

Conclusion: Biomarkers are valuable tools in diagnosing and managing prostate cancer. Utilizing a combination of several biomarkers may improve diagnostic accuracy and predict treatment outcomes. Future research should focus on developing new biomarkers and optimizing diagnostic methods to achieve better results in managing this disease.

Keywords: Prostate cancer, Biomarkers, PSA, PCA3, kallikrein markers

The Role of Gut Microbiota in Modulating Immune Responses and Cancer Therapy Efficacy (Review)

Mohammad Hossein Khazaee-Nasirabadi,^1,*

1. Department of Medical Laboratory Sciences, School of Allied Medical Sciences, Jiroft University of Medical Sciences, Jiroft, Iran

Introduction: The gut microbiota influences host immunity, which in turn controls the gut microbiome to preserve homeostasis. By influencing immunological regulation, systemic metabolism, and circulation, the gut microbiota can impact not just gut immunity but also immune responses in distal mucosal regions. Additionally, microbial immunomodulatory metabolites and immune responses are correlated, indicating potential therapeutic manipulation of tumors in the future.

Methods: The analysis of publications that were available in the PubMed and Web of Sciences databases served as the foundation for the results of the investigation. We identified this research using keywords such as gut microbiome, immune modulation, immunotherapy, antibiotics, probiotics, fecal microbiota transplantation.

Results: of the byproducts of microbial fermentation of fiber, short-chain fatty acids (SCFAs) facilitate colonic Treg expansion by suppressing histone deacetylase (HDAC) activity and targeting Forkhead box P3 (FOXP3), the major nuclear transcription factor of Tregs. Tregs either exhibit anti-inflammatory effects against carcinogenesis or mitigate anticancer responses when they infiltrate the TME. Other microbial metabolites, including polyamines, suppress anticancer immunity by causing the production of tumor-derived proteases, which increase the invasiveness of tumor cells, and by preventing lymphocyte proliferation.

Conclusion: Immunotherapy and the immune response to cancer are significantly influenced by the gut microbiota. Thus, it has been suggested that the best use of microbial therapy in the treatment of cancer is microbiota precision medicine, which includes, prebiotics, probiotics, antibiotics, and vaccines.

Keywords: gut microbiome, immune modulation, immunotherapy

the role of radiation on tumorigenesis (Review)

Farzane Hosseinzade,^1,*

1. Varastegan institute for medical sciences

Introduction: Radiation, particularly ionizing radiation, is a well-established risk factor for various malignancies. While the carcinogenic potential of non-ionizing radiation remains inconclusive, its role in cellular damage, especially in the context of prolonged exposure, is an ongoing area of research. Individuals undergoing radiotherapy, radiologists, and miners are exposed to significant doses of ionizing radiation, increasing their risk of developing secondary malignancies. In contrast, non-ionizing radiation, such as that emitted by mobile phones, primarily induces thermal effects in tissues. However, studies have indicated a potential negative impact on sperm motility, highlighting the need for further investigation into the long-term effects of such exposure.

Methods: In this study, out of 72 primary articles searched in PubMed and Google Scholar databases , 12 articles with the keywords Radiation, Malignancy, Radiotherapy,DNA damage ,and Infertility were selected and studied.

Results: Exposure to ionizing radiation can trigger the formation of highly reactive molecules known as reactive oxygen species (ROS) within cells. These ROS can inflict significant damage on critical cellular components, including DNA, proteins, and lipids. Among the various types of cellular damage caused by ROS, DNA damage is particularly concerning. Double-strand breaks (DSBs) can lead to genomic instability and cell death. If not repaired accurately, these breaks can result in mutations, which may contribute to the development of cancer.

Conclusion: To mitigate the adverse health effects of ionizing radiation, it is imperative to minimize exposure. For individuals occupationally exposed to radiation, rigorous monitoring of cumulative dose is essential. In cases of excessive exposure, strategies such as extended leave can be implemented to reduce accumulated radiation dose. Furthermore, Individuals carrying specific genetic variants, such as ATM, exhibit increased risk of radiation-induced malignancies. Therefore, pre-employment genetic screening can be a valuable tool for identifying individuals who may be at heightened risk and implementing appropriate protective measures.

Keywords: Radiation, Malignancy, Radiotherapy, DNA damage , Infertility

Thioredoxin Peroxidase: A Target for Vaccine Development and Serological Detection of Echinococcosis (Research Paper)

Roohollah Fattahi,^1,*

1. Department of laboratory and clinical sciences, Faculty of veterinary sciences, Ilam University, Ilam, Iran

Introduction: Human cystic echinococcosis (CE) is a chronic infection caused by the cestode Echinococcus granulosus. The robust humoral and cellular immune response elicited by this parasite. This study investigates the physicochemical and antigenic properties of the Thioredoxin peroxidase protein (XP_024356136.1) utilizing bioinformatics tools, aiming to evaluate its viability as a candidate for vaccine development and serological detection of Echinococcus granulosus across diverse host species.

Methods: The antigenicity, allergenicity, and toxicity of the TP protein were assessed using ANTIGENpro, AllerTop, and ToxinPred servers, respectively. The ProtParam server was employed to analyze the physicochemical characteristics of the protein. Additionally, linear B-cell and cytotoxic T lymphocyte (CTL) epitopes were predicted using the ABCpred webserver and the IEDB database, contributing to a comprehensive understanding of the protein's potential in immunological applications.

Results: The analysis demonstrated that the thioredoxin peroxidase (TP) protein is antigenic, with an ANTIGENpro score of 0.672609, and is classified as non-allergenic and non-toxic. Physicochemical characterization revealed a molecular weight of 21,405.51 Daltons and an estimated half-life of 30 hours in mammalian reticulocytes in vitro, over 20 hours in yeast in vivo, and more than 10 hours in Escherichia coli in vivo. The instability index was calculated to be 30.19, indicating favorable stability for vaccine formulation. Additionally, the aliphatic index of 85.39 classifies the protein as thermostable, while the theoretical isoelectric point (pI) was determined to be 5.79. The GRAVY index of -0.093 suggests a polar nature with significant water interaction, indicating high solubility. The protein comprises 24 negatively charged residues (aspartic acid and glutamic acid) and 21 positively charged residues (arginine and lysine), with a molecular formula of C954H1487N257O281S11 and a total atom count of 2,990. The QuerySol scaled solubility value was determined to be 0.455.

Conclusion: Furthermore, our findings identified multiple linear B-cell and cytotoxic T lymphocyte (CTL) epitopes within the TP protein, all exhibiting high antigenicity, thereby highlighting their potential for application in vaccine development and serological detection of Echinococcus granulosus.

Keywords: Echinococcus granulosus, Thioredoxin peroxidase, epitopes

Transmission routes and differential diagnoses of HPV infection (Review)

Masoud Lahouty,^1,* Fatemeh Ramzi,² Mohadeseh Ramzi,³

1. Pediatric Health Research Center, Tabriz University of Medical Sciences, Tabriz iran.
2. Department of Microbiology and Virology, School of Medicine, Urmia University of Medical Sciences, Urmia, Iran.
3. Department of Medical Parasitolog and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

Introduction: HPV is the most common viral disease of the genital tract. This virus has more than 250 serotypes. This virus is one of the main causes of cervical cancer in women and penile cancer in men. On the other hand, it causes psychological damage and depression in people infected with this virus. This virus is transmitted through anal-oral intercourse in 80% of cases and through small skin wounds and scratches in others. Note that a competent immune system is crucial in preventing infection with this virus. Given that several lesions resulting from skin allergies and allergic reactions in the immune system are very similar to those resulting from HPV, timely diagnosis and identification of this infection are of enormous importance in the transmission and prevention of diseases and cancer. Therefore, the purpose of this study is to investigate the differential diagnosis of lesions resulting from infection with this virus with other genital lesions.

Methods: We conducted a study using keywords related to sexually transmitted diseases (STDs), papillomavirus, and HPV risk factors from a variety of databases such as PubMed, Google Scholar, and Sid.ir. Finally, among other resources, we used data from 2015 to 2024. We selected 20 articles, studied them, and conducted a comprehensive analysis.

Results: An accurate and timely diagnosis of HPV-related skin lesions is essential for timely treatment and prevention of virus transmission. A number of genital lesions resemble lesions caused by this virus, including Candidioma, skin tags, keratosis pilaris, Fordyce angiomas, papillae coronae glands, and epidermoid cysts. None of the lesions are contagious or transmissible. For example, Candidioma, which is an allergic reaction to theyeast Candida albicans that appears on the penis during sexual intercourse with a woman who has a vaginal yeast infection, closely resembles HPV lesions and isnot transmissible. Or, as well, pearly papules, which closely resemble HPV lesions. Therefore, a specialist physician and laboratory tests are required for the correct diagnosis and treatment of HPV.

Conclusion: Oral sex, especially oral-anal intercourse, carries the highest risk of infection with this virus. Using condoms, avoiding oral sex, and getting vaccinated against HPV are among the most important ways to combat infection with this virus. The best laboratory diagnostic test is the PCR molecular test, which even determines the serotype of this virus, which is crucial for treatment. The tissue tropism and location of HPV lesions on the genitals and penis are different from other lesions, but this diagnosis is not definitive and requires high experience.

Keywords: HPV, human papillomavirus, HPV Transmission, cancer, HPV diagnosis

Unveiling Autoimmune Responses in Paraneoplastic Syndromes: Diagnostic Challenges and Clinical Implications (Review)

Mobina Bagherian,¹ Mahdi Moassesfar,^2,* Hadiseh Vahabi,³

1. Department of Laboratory Science, Faculty of Paramedical Sciences, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
2. Department of Biology, Faculty of Biological Sciences, North Tehran Branch, Islamic Azad University, Tehran, Iran.
3. Student Research Committee, Department of Medical Laboratory Sciences, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran

Introduction: Paraneoplastic syndromes (PNS) are complex, rare conditions that arise as a result of the immune system's response to tumors. These syndromes often involve neurological manifestations, such as peripheral neuropathy, cerebellar ataxia, and encephalomyelitis, which are triggered by immune responses against shared antigens between cancer cells and normal tissues. Key antibodies such as anti-Hu, anti-Yo, and anti-CRMP5 are crucial for diagnosing PNS. However, these antibodies are not always present, and their variability complicates accurate diagnosis, often delaying treatment. This review explores the immune mechanisms involved, the role of onconeural antibodies, and the challenges in diagnosing PNS early enough to improve patient outcomes.

Methods: A literature review was conducted using PubMed and Google Scholar. Peer-reviewed articles in English that specifically addressed autoimmune PNS, diagnostic approaches, and antibody associations were included. Studies unrelated to paraneoplastic etiologies were excluded.

Results: Recent studies have established strong associations between specific antibodies and various cancer types. Anti-Hu antibodies are prominently linked to small cell lung cancer (SCLC) and are often associated with paraneoplastic encephalomyelitis. Anti-Yo antibodies are predominantly found in women with gynecological malignancies, particularly ovarian and breast cancers, and are commonly linked to cerebellar degeneration. Anti-CRMP5 antibodies are associated with lung cancer and are frequently identified in patients presenting with peripheral neuropathy and cerebellar ataxia, making this antibody significant in these cases. Anti-Ri antibodies are observed in patients with breast and lung cancers and are typically linked to opsoclonus-myoclonus syndrome paraneoplastic cerebellar degeneration. Advancements in imaging technologies, such as PET-CT, have enhanced the detection of underlying malignancies in patients with paraneoplastic syndromes. The combination of imaging with antibody profiling has proven effective in uncovering associated tumors. Nevertheless, some cases remain challenging due to the absence of identifiable antibodies, indicating the need for improved diagnostic approaches.

Conclusion: Paraneoplastic syndromes remain a diagnostic challenge due to their rarity, diverse clinical presentations, and incomplete antibody detection. The immune responses against shared tumor and host antigens play a crucial role in the development of PNS, often serving as early indicators of malignancy. Despite progress in diagnostic tools, such as antibody testing and advanced imaging, a substantial number of cases still go undiagnosed or misdiagnosed. A multidisciplinary approach combining clinical evaluation, advanced imaging, and serological testing is essential for accurate diagnosis and timely intervention. Future research should focus on identifying new biomarkers and refining existing diagnostic methods to improve early detection and treatment outcomes for patients with PNS.

Keywords: Paraneoplastic syndromes, Autoimmune response, Onconeural antibodies, Cancer diagnosis.

VHL Disease and p.Gly93 Amino Acid Disorder (Review)

Issa Layali,¹ Baran Nekoee Seresht,^2,* Ali Pourseif,³

1. Department of Biochemistry and Biophysics, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
2. Department of Cellular and Molecular Biology, Faculty of Advanced Sciences and Technology, Islamic Azad University of Medical Sciences, Tehran, Iran
3. Department of Immunology, Islamic Azad University of Medical Sciences, Tehran, Iran

Introduction: Von Hippel-Lindau (VHL) disease is a rare genetic disorder characterized by an increased risk of various tumors due to a mutation in the VHL gene, which encodes a protein of the same name. The VHL protein plays a critical role in regulating cellular signaling pathways, angiogenesis, and apoptosis. In this review article, we will focus on the biochemical and immunological aspects of VHL disease, examining the role of the protein in these processes, mechanisms of tumor formation, and recent advances in treating the disease. Biochemical Metabolism Perspective on VHL Disease: - VHL Gene and Protein: The VHL gene is located on chromosome 3, encoding a protein that is part of a larger protein complex known as E3 ubiquitin ligase, which functions in the degradation of specific proteins, including the hypoxia-inducible factor (HIF) transcription factor. - HIF Pathway: HIF is a transcription factor activated in response to reduced oxygen levels. Under normal conditions, the VHL protein identifies and binds to HIF, leading to its degradation. Consequently, the expression of HIF-regulated genes, such as those involved in angiogenesis and cellular metabolism, is reduced. - Role of VHL in Tumorigenesis: A mutation in the VHL gene disrupts VHL protein function, resulting in the sustained elevation of HIF levels. Elevated HIF activates HIF target genes excessively, ultimately promoting tumor growth. In-Depth Insight: VHL disease is a rare genetic disorder characterized by tumor and cyst formation in various organs, due to mutations in the VHL gene, which encodes the tumor suppressor VHL protein. This protein is crucial for regulating cellular signaling pathways, angiogenesis, and cellular metabolism. Role of VHL Protein in Cellular Metabolism: - Regulation of HIF Pathway: The VHL protein acts as a component of the E3 ubiquitin ligase complex, targeting hypoxia-inducible factor (HIF) for degradation by the proteasome. HIF stimulates genes involved in angiogenesis, glycolytic metabolism, and cell survival. - Amino Acid Metabolism Regulation: VHL protein is also involved in amino acid metabolism regulation, as mutations in the VHL gene are associated with branched-chain amino acid (BCAA) metabolism disruption. - Carbohydrate Metabolism Regulation: VHL protein regulates carbohydrate metabolism as well, with studies linking VHL gene mutations to increased glycolytic enzyme activity and lactate production. These metabolic shifts help tumor cells survive and proliferate under hypoxic conditions. Metabolic Disorders and Tumor Growth in VHL Disease: - Increased Glycolysis: Tumor cells in VHL disease often rely on glycolysis for energy, even in the presence of sufficient oxygen, a phenomenon known as the Warburg effect. - Increased Amino Acid Synthesis: VHL disease tumor cells require significant amino acids for rapid protein synthesis and growth, one of their distinguishing features. - Increased Nucleotide Production: Tumor cells also need nucleotides in abundance for DNA replication and cell division. What is HIF? - HIF: Hypoxia-Inducible Factor, proteins that play an essential role in cellular response to oxygen deprivation. Functions: When oxygen levels decrease, HIFs activate specific genes that help cells adapt to low oxygen, including promoting blood vessel formation, changing metabolism, and stimulating cell growth. - Significance: HIF proteins are involved in embryonic development, response to tissue damage, and tumor growth. Therapeutic Outlook: Understanding the molecular and metabolic mechanisms of VHL disease enables the development of new treatments. Potential therapeutic approaches include: - HIF Inhibitors: Drugs that inhibit HIF activity to slow tumor growth. - Glycolysis Inhibitors: These drugs inhibit the glycolytic pathway, disrupting tumor cell energy supply. - Amino Acid Synthesis Inhibitors: Drugs that inhibit amino acid synthesis limit tumor cell growth. - Gene Therapy: This approach involves replacing the mutated VHL gene to restore normal cellular function.

Methods: VHL disease is a complex disorder in which metabolic abnormalities significantly contribute to tumor growth. By improving our understanding of these mechanisms, we can develop more effective, personalized treatments for patients with VHL.

Results: Immunology of VHL Disease: - Immune System and Tumors: The immune system can normally identify and eliminate tumor cells. However, tumor cells use various mechanisms to evade the immune response. - Surgical Treatment: Tumor removal surgery is a primary treatment for this disease. - Drug Therapy: Medications such as angiogenesis inhibitors and mTOR inhibitors are used to treat VHL disease. - Immunotherapy: Immunotherapy, using CAR T-cells and immune checkpoint inhibitors, is an emerging approach in treating VHL

Conclusion: VHL disease is a complex genetic disorder. Understanding the molecular mechanisms underlying VHL disease is essential for developing effective treatments. Recent studies have significantly advanced our understanding of the role of VHL protein in regulating cellular signaling pathways and tumorigenesis. Given the immune system's importance in cancer treatment, immunotherapy has emerged as a promising approach for treating VHL disease.

Keywords: VHL disease, VHL gene, VHL protein, HIF, tumor, angiogenesis, apoptosis, immunotherapy

Viral vector-based gene therapies in the clinic; a review article. (Review)

Ayda Khatibi,^1,*

1. Department of Biological Sciences, Faculty of Basic Sciences, University of Nabi Akram, Tabriz.

Introduction: Gene therapy, which involves the modification of gene expression or the correction of dysfunctional genes, presents significant potential as a therapeutic approach for the treatment of a wide range of diseases. Unlike conventional pharmaceuticals, gene therapy involves the genetic modification of cells, thereby creating avenues for the potential treatment of diseases once deemed incurable. The concept of gene therapy can be traced back to the 1960s, when initial investigations demonstrated the potential to introduce DNA sequences into mammalian cells for the purpose of gene repair. Over several decades, significant scientific advancements culminated in the first human gene therapy clinical trial conducted in 1990, which utilized retrovirus vector technology to treat severe combined immunodeficiency. However, the field encountered a substantial setback in the late 1990s, following the death of a patient due to immune reactions triggered by the viral vector in a 1999 trial. Additionally, the emergence of viral vector-induced leukemia in four patients who received retrovirus-based gene therapy in another trial in 2000 raised serious safety concerns. These incidents temporarily halted clinical applications of gene therapy and underscored the urgent need for the development of safer viral vectors. In the ensuing decade, research efforts concentrated on enhancing the understanding of the biology of viral vectors, as well as on advancing the engineering of safer and more effective vector technologies. This progress ultimately led to the first clinical approvals of gene therapy products. Notably, China granted approval for the world’s first gene therapy product, Gendicine, for the treatment of head and neck cancer in 2003. The European Medicines Agency (EMA) followed with the first approval of a gene therapy product in 2012 (Glybera), and the United States approved its inaugural gene therapy product in 2017 (Kymriah). Gene therapies are designed to introduce genetic material into target cells, representing a promising approach to the treatment of diseases previously regarded as incurable by conventional methods. In numerous instances, gene therapy necessitates the use of a vector to facilitate the delivery of gene therapeutics into target cells. Among the various types of vectors, viral vectors are extensively studied due to their notable advantages, including exceptional transduction efficiency. Over several decades, the development of viral vector-based gene therapies has yielded promising clinical outcomes, with numerous products receiving approval for the treatment of various diseases, including cancer, infectious diseases, and monogenic disorders. Furthermore, a significant number of active clinical trials are currently being conducted to explore and expand the therapeutic potential of these innovative treatments. In this review, we emphasize the diverse range of viral vectors and examine currently approved products while outlining the existing clinical landscape of in vivo viral vector-based gene therapies. Furthermore, we provide a critical analysis of the primary translational challenges associated with these therapies and discuss potential strategies to address these issues effectively.

Methods: In our research, we conducted an extensive examination of articles sourced from several reputable databases, including Scopus, PubMed, Google Scholar, Civilica, and ScienceDirect. Our search was centered around specific terms: gene therapy, modifying expression, vectors, and vector-based gene therapies pertaining to neurological diseases. We placed particular emphasis on recent publications and their corresponding references as essential sources for our study. Furthermore, our search parameters were limited to articles published in English and Persian.

Results: Because it is a review article, the results and conclusions are brought together.

Conclusion: Gene therapy represents a groundbreaking approach to the treatment of diseases by directly correcting or modifying genes. This innovative modality holds a distinctive place within the spectrum of therapeutics, as it offers the potential to cure certain diseases, notably genetic disorders. Numerous gene therapy products, particularly those utilizing viral vectors, have attained clinical approval for the treatment of a diverse range of diseases. These include not only genetic disorders but also other conditions such as cancer and infectious diseases. Recent approvals of adeno-associated virus (AAV)-based products have significantly transformed the field of gene therapy, providing the first viable treatment options for monogenic disorders. Furthermore, an increasing number of active clinical trials are emerging, focused on investigating novel viral vector-based gene therapies within the clinical landscape. The ongoing clinical translation of viral gene therapies is encountering substantial challenges related to manufacturing, biological considerations, and regulatory compliance. Addressing these challenges necessitates considerable efforts across preclinical, clinical, and commercial domains. Nevertheless, recent scientific advancements in viral vector engineering, genomic disease identification, and foundational gene editing technologies are ushering viral gene therapies into a new era. It is anticipated that viral vector-based gene therapy will remain an active focus in clinical settings, with an increasing number of gene therapy products expected to enter the market in the foreseeable future.

Keywords: Gene therapy, Modifying Expression, Vectors, Vector-based gene therapies.

Vitamin D deficiency as a debatable modulator of thalassemia major outcomes; an overview of the latest findings (Review)

Omolbanin Sargazi Aval,^1,* Ali Bazi,² Erfan Ebrahimi,³

1. Department of Hematology, Faculty of Allied Medical Sciences, Zabol University of Medical Sciences, Zabol, Iran.
2. Department of Hematology, Kerman University of Medical Sciences, Kerman, Iran
3. Student Research Committee, Zabol University of Medical Sciences, ZaboL, Iran.

Introduction: The role of vitamin D, and more importantly, vitamin D deficiency (VDD), in thalassemia clinical course is controversial. Vitamin D is essential for bone metabolism and calcium turnover, and patients with transfusion-dependent thalassemia (TDT) are generally supplemented with vitamin D to prevent VDD, yet this phenomenon seems to be inevitable in these patients as they grow. The direct and independent association of VDD with bone mineral density and osteoporosis in TDT patients is debated as several other modulators seem to interfere with this pathway. Regardless of chelation and transfusion therapy history or receiving supplementations, aging seems to be the most important predictor of VDD in these patients, and other players can either accelerate to postpone its occurrence.

Methods: Using keywords thalassemia, vitamin D, vitamin D deficiency, siderosis, hemosiderosis, osteoporosis, cardiomyopathy, and cardiac disfunction, relevant studies were obtained in PubMed, Google Scholar (100 first records), Science Direct, Cohcrane library, Springer, Web of Science, and Wiley Online Library.

Results: Attention should be paid to VDD in older TDT patients regarding the recent findings suggesting a role for VDD in predisposing to cardiac iron deposition, which is thought to be mediated through L-type voltage-dependent calcium channels, which are expressed on cardiomyocytes and serve as the main entry route of calcium and, probably, non-transferrin bound iron, into cardiac muscles.

Conclusion: Due to the fact that cardiomyopathy is the main cause of death in older TDT patients, correcting cardiac dysfunction in these patients may require resolving underlying VDD. Here, we reviewed the risk factors and important clinical implications of VDD in TDT patients.

Keywords: Thalassemia, Vitamin D, Metabolism, Cardiac dysfunction, Bone disorders