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Background and purpose: The global emergence of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has prompted widespread concern. Bacteriophages have recently gained attention as a cost-effective and stable alternative for vaccine development due to their adjuvant properties. This study aimed to design and validate a poly epitope composed of viral proteins. Experimental approach: SARS-CoV-2 proteins (spike, nucleocapsid, membrane, envelope, papain-like protease, and RNA-dependent RNA polymerase) were selected for analysis. Immunoinformatic methods were employed to predict B and T cell epitopes, assessing their antigenicity, allergenicity, and toxicity. Epitopes meeting criteria for high antigenicity, non-allergenicity, and non-toxicity were linked to form poly epitopes. These sequences were synthesized and cloned into pHEN4 plasmids to generate Poly1 and Poly2 phagemid vectors. Recombinant Poly1 and Poly2 phages were produced by transforming M13ΔIII plasmids and phagemid vectors into E. coli TG1. Female Balb/c mice were immunized with a cocktail of Poly1 and Poly2 phages, and their serum was collected for ELISA testing. Interferon-gamma (IFN-γ) testing was performed on spleen-derived lymphocytes to evaluate immune system activation. Findings/Results: Recombinant Poly1 and Poly2 phages were produced, and their titer was determined as 1013 PFU/mL. Efficient humoral immune responses and cellular immunity activation in mice were achieved following phage administration. Conclusion and implication: Poly epitopes displayed on phages exhibit adjuvant properties, enhancing humoral and cellular immunity in mice. This suggests that phages could serve as adjuvants to bolster immunity against SARS-Cov-2. Recombinant phages could be applied as effective candidates for injectable and oral vaccine development strategies.

Abstract Background Human dental pulp-derived mesenchymal stem cells (hDP-MSCs), which include human dental pulp stem cells (hDPSCs) and stem cells from human exfoliated deciduous teeth (SHEDs), are promising cell sources for regenerative therapies. Nevertheless, a lack of knowledge relating to the mechanisms regulating their differentiation has limited their clinical application. microRNAs (miRNAs) are important regulatory molecules in cellular processes including cell differentiation. This systematic review aims to provide a panel of miRNAs that regulate the differentiation of hDP-MSCs including hDPSCs and SHEDs. Additionally, bioinformatic analyses were conducted to discover target genes, signaling pathways and gene ontologies associated with the identified miRNAs. Methods A literature search was performed in MEDLINE (via PubMed), Web of Science, Scopus, Embase and Cochrane Library. Experimental studies assessing the promotive/suppressive effect of miRNAs on the differentiation of hDP-MSCs and studies evaluating changes to the expression of miRNAs during the differentiation of hDP-MSCs were included. miRNAs involved in odontogenic/osteogenic differentiation were then included in a bioinformatic analysis. A miRNA-mRNA network was constructed, and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed. A protein–protein interaction (PPI) network was also constructed. Results Of 766 initially identified records through database searching, 42 and 36 studies were included in qualitative synthesis and bioinformatic analyses, respectively. Thirteen miRNAs promoted and 17 suppressed odontogenic/osteogenic differentiation of hDP-MSCs. hsa-miR-140-5p, hsa-miR-218 and hsa-miR-143 were more frequently reported suppressing the odontogenic/osteogenic differentiation of hDP-MSCs. hsa-miR-221 and hsa-miR-124 promoted and hsa-miR-140-5p inhibited neuronal differentiation, hsa-miR-26a-5p promoted and hsa-miR-424 suppressed angiogenic differentiation, and hsa-miR-135 and hsa-miR-143 inhibited differentiation within myogenic lineages. A miRNA-mRNA network including 1890 nodes and 2171 edges was constructed. KEGG pathway analysis revealed MAPK, PI3K-Akt and FoxO as key signaling pathways involved in the odontogenic/osteogenic differentiation of hDP-MSCs. Conclusions The findings of this systematic review support the potential application of the specific miRNAs to regulate the directed differentiation of hDP-MSCs in the field of regenerative therapies.

Objective(s): Hepatocellular carcinoma (HCC) is a common and lethal type of cancer worldwide. The importance of non-coding RNAs such as long non-coding RNAs (lncRNAs), circular RNAs (circRNAs), and microRNAs (miRNAs) have been recognized in the development of HCC. In this study, we constructed a four-component competing endogenous RNA (ceRNA) network in HCC and evaluated prognostic values of the ceRNAs. Materials and Methods: The expression profiles of lncRNAs, miRNAs, and mRNAs were retrieved from The Cancer Genome Atlas database. GSE94508 and GSE97332 studies from the Gene Expression Omnibus database were used to identify circRNAs expression profiles. A four-component ceRNA network was constructed based on differentially-expressed RNAs. Survival R package was utilized to identify potential prognostic biomarkers.Results: A four-component ceRNA network including 295 edges and 239 nodes was constructed and enrichment analysis revealed important Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways. A Protein-Protein Interaction network with 118 nodes and 301 edges was also established. The enhancer of zeste homolog 2 (EZH2) was the highest degree hub gene in the PPI network. Because of the significance of EZH2 in HCC, we presented its axes in the ceRNA network, which play important roles in HCC progression. Furthermore, ceRNAs were identified as potential prognostic biomarkers utilizing survival analysis.Conclusion: Our study elucidates the role of ceRNAs and their regulatory interactions in the pathogenesis of HCC and identifies EZH2-related RNAs which may be utilized as novel therapeutic targets and prognostic biomarkers in the future.

Objective: Given the prevalence of fertility problems in couples and the defect in embryo implantation as well as the low success rate of assisted reproductive techniques, it is necessary to investigate the causes of this phenomenon. Type 2 diabetes mellitus (T2DM) is a metabolic disease with multiple effects on various organs as well as the endometrium. In this study, the effects of endometrial cell culture on the expression of α3 and β1 integrin genes and protein in type 2 diabetic rats were investigated. Materials and Methods: In this experimental study, 35 female rats were divided into five groups: control, sham, diabetic, Pioglitazone-treated and Metformin-treated groups. First, rats were maintained in diabetic condition for 4 weeks. Then, treatment was performed for the next four weeks. Four weeks after induction of diabetes, rats were sacrificed at the time of embryo implantation. The uterus was removed. Endometrial cells were isolated and cultured for 7 days. Immunocytochemistry staining was used to confirm endometrial cells. Expression of α3 and β1 integrin genes was determined by real-time polymerase chain reaction (PCR) technique and the α3β1 protein content measured using Western blot both before and after endometrial cell culture. Result: The expression level of α3 integrin gene in the Pioglitazone-treated group compared with metformin-treated group was significantly decreased (P

Abstract Sandhoff disease is one of the GM2‐gangliosidoses which is caused by a mutation in the HEXB preventing the breakdown of GM2‐ganglioside. We report a novel HEXB variant in a family with a history of a dead girl with Sandhoff disease which was not found in controls.

Background: Mucin-1(Muc1) is one of the first molecules in the endometrium that confronts implanting embryos.There is insufficient knowledge about the impacts of diabetes and drugs developed for diabetes treatment on expressionof this molecule at the time of implantation. Therefore, this study aimed to investigate the impacts of diabetes andinsulin, metformin and pioglitazone on Muc1 expression at the time of implantation.Materials and Methods: This experimental study was conducted on a total of 63 female Wistar rats divided into 9groups. To induce type 1diabetes, streptozotocin (STZ) and for induction of type 2 diabetes, nicotinamide (NA) andSTZ were injected intraperitoneally. For superovulation, human menopausal gonadotropin (HMG) and human chorionicgonadotropin (HCG) were used. Insulin, metformin and pioglitazone were administered for two weeks. Finally,the endometrial expression of Muc1 was evaluated by quantitative real-time reverse transcription–polymerase chainreaction (RT-PCR).Results: Muc1 expression was non-significantly increased in type 1 and type 2 diabetic groups compared to the controlgroup (P=0.61 and 0.13, respectively); also, it increased in insulin-treated type 1 diabetic group compared to thecontrol group (P=0.0001). Its expression was increased in insulin-treated type 1 diabetic group compared to untreateddiabetic group (P=0.001). The expression level of Muc1 was significantly reduced in superovulated and insulin-treatedtype 1 diabetic group compared to the insulin-treated type 1 diabetic group (P=0.001).Conclusion: One of the causes of fertility problems in diabetes, is changes in Muc1 expression during implantation.On the other hand, the use of insulin in these patients can even lead to overexpression of this gene and worsen thecondition. However, these changes can be partially mitigated by assisted reproductive technology (ART) such as superovulation.Also, treatment with metformin and pioglitazone can restore Muc1 expression to near normal levels andhas beneficial effects on implantation

Background: Implantation requires intimate crosstalk between the embryo and uterus for a successful establishment of pregnancy. Type 2 diabetes mellitus may lead to implantation failure. The effect of diabetes and its therapeutic drugs on implantation is still largely unclear. Objective: To assess the endometrial expression changes of vascular endothelial growth factor A (VEGFA) and leukemia inhibitory factor (LIF), at the time of implantation in diabetic rats following treatment with Metformin and Pioglitazone. Materials and Methods: Twenty-eight 6-8-wk-old Wistar female rats weighing 200-250 gr were divided into four groups (n = 7/each). Type 2 diabetes was induced and Metformin and Pioglitazone were applied for 4 wk. The expression of VEGFA and LIF was measured by real-time reverse transcription-polymerase chain reaction and Western blot. Results: The relative expression of VEGFA transcript was higher in the diabetic (p = 0.02) and Metformin-treated (p = 0.04) rats compared to the control group. Furthermore, the VEGFA transcript level significantly reduced in Pioglitazone-treated diabetic rats (p = 0.03). LIF expression was elevated in the Metformin- and the Pioglitazone-treated rats and reduced in the diabetic group in comparison with the control group. Compared to the diabetic rats, the expression of LIF was significantly elevated in the Metformin- (p = 0.01) and Pioglitazone-treated (p = 0.03) groups. Conclusion: The expressions of LIF and VEGFA were altered in diabetic rats during implantation which may be associated with diabetic-related infertility. Pioglitazone is able to restore the VEGFA and LIF expressions to their baseline levels more efficiently than Metformin.Key words: Embryo implantation, Leukemia inhibitory factor, Vascular endothelial growth factor A, Metformin, Pioglitazone, Rats.}

Background: Gastric cancer is a worldwide life-threatening cancer. The underlying cause of it is still unknown. We have noticed that some cancer/testis antigens (CTAs) are up-regulated in gastric cancer. The role of these genes in gastric cancer development is not fully understood. The main aim of the current study was to comprehensively investigate CTAs' expression and function in stomach adenocarcinoma (STAD). Materials and Methods: A comprehensive list of CTA genes was compiled from different databases. Transcriptome profiles of STAD were downloaded from the cancer genome atlas (TCGA) database and analyzed. Differentially-expressed CTAs were identified. Pathway enrichment analysis, weighted gene correlation network analysis (WGCNA), and overall survival (OS) analysis were performed on differentially-expressed CTA genes. Results: Pathway enrichment analysis indicates that CTA genes are involved in protein binding, ribonucleic acid processing, and reproductive tissues. WGCNA showed that six differentially-expressed CTA genes, namely Melanoma antigen gene (MAGE) family member A3, A6, A12 and chondrosarcoma associated gene (CSAG) 1, 2, and 3, were correlated. Up-regulation of MAGEA11, MAGEC3, Per ARNT SIM domain containing 1 (PASD1), placenta-specific protein 1 (PLAC1) and sperm protein associated with the nucleus X-linked family member (SPANXB1) were significantly associated with lower OS of patients. Conclusion: MAGEA11, MAGEC3, PASD1, PLAC1, and SPANXB1 can be investigated as prognostic biomarkers in basic and clinical studies. Further functional experiments are needed to understand the exact interaction mechanisms of these genes.

Gastric cancer is one of the most common cancers in the world. Late diagnosis is the main cause of the high rate of treatment failure and death among patients with gastric cancer; therefore identifying the molecular basis of cancer initiation and metastasis is so critical for developing efficient methods for early diagnosis and therapy. long non-coding RNAs (lncRNAs) are the largest group of noncoding RNAs, which are involved in cancer pathogenesis. Regarding the roles of lncRNA-HNF1A-AS1 and lncRNA-MVIH in cancer pathogenesis and progression, we aimed to evaluate expression profiles and clinicopathological relevance of these two genes in human gastric cancer. Real-time PCR was performed to assess relative gene expressions in 60 tumoral and non-tumoral gastric tissues. The association between gene expressions and clinicopathological characteristics were also analyzed. Expression and clinicopathological data of the lncRNA-HNF1A-AS1 from 318 gastric cancer patients were also downloaded from the TCGA database. HNF1A-AS1 was down-regulated in GC tissues and MVIH did not show any significant differential expression in GC tissues. Otherwise, the expression of lncRNA-HNF1A-AS1 was significantly higher in TCGA tumor samples. Furthermore, lncRNA-HNF1A-AS1 expression was associated with tumor metastasis and that of lncRNA-MVIH showed association with tumor grade and stage. lncRNA-HNF1A-AS1 expression status did not show any significant correlation with GC overall survival. In conclusion, lncRNA-HNF1A-S1 and lncRNA-MVIH genes may play a critical role in gastric cancer progression. Functional studies on the mechanism of action of these two lncRNAs could help to understand their role in gastric cancer progression.

Background The present study was designed to evaluate serum lipid profile and tumor necrosis factor-alpha (TNF-ɑ) level in diabetic rats at implantation time. Type 2 diabetes mellitus (T2DM) could affect various systems, including innate immune system and it causes chronic low-grade inflammation, increasing level of TNF-ɑ. Furthermore, T2DM is often accompanied by impaired lipid profile. Metformin and pioglitazone are used as the first and second lines of treatment for T2DM. Materials and Methods In this experimental study, 35 adult virgin female wistar rats, weighting 175-225 g, were randomly categorized into five groups: i. Control, ii. Sham, iii. Nicotinamide (NA)+streptozotocin (STZ) induced T2DM, iv. Diabetic+pioglitazone (20 mg/kg/day for 28 days oral administration), and v. Diabetic+metformin (100 mg/kg/day for 28 days oral administration). At the time of implantation, TNF-ɑ level in serum of rats was measured by ELISA kit. Glucose was measured using photometric method and lipid profiles were calculated by enzymatic methods. Results Level of TNF-ɑ in the diabetic group was significantly higher than other groups (P < 0.001). In metformin treated group, TNF-ɑ serum level was also significantly higher than pioglitazone treated group (P < 0.001). Fasting blood sugar (FBS) and lipid profiles were significantly higher in diabetic group. Conclusion Metformin and pioglitazone have similar effects on glucose, lipid profiles and TNF-ɑ serum levels. Among these drugs, pioglitazone has more efficient influence on TNF-α serum level, in comparison with metformin.

Abstract Background Diabetes mellitus deeply changes the genes expression of integrin (Itg) subunits in several cells and tissues such as monocytes, arterial endothelium, kidney glomerular cells, retina. Furthermore, hyperglycemia could impress and reduce the rate of successful assisted as well as non-assisted pregnancy. Endometrium undergoes thorough changes in normal menstrual cycle and the question is: What happens in the endometrium under diabetic condition? Objective The aim of the current study was to investigate the endometrial gene expression of α3, α4, αv, Itg β1 and β3 subunits in diabetic rat models at the time of embryo implantation. Materials and Methods Twenty-eight rats were randomly divided into 4 groups: control group, diabetic group, pioglitazone-treated group, and metformin-treated group. Real-time PCR was performed to determine changes in the expression of Itg α3, α4, αv, β1, and β3 genes in rat's endometrium. Results The expression of all Itg subunits increased significantly in diabetic rats' endometrium compared with control group. Treatment with pioglitazone significantly reduced the level of Itg subunits gene expression compared with diabetic rats. While metformin had a different effect on α3 and α4 and elevated these two subunits gene expression. Conclusion Diabetes mellitus significantly increased the expression of studied Itg subunits, therefore untreated diabetes could be potentially assumed as one of the preliminary elements in embryo implantation failure.

Background Osteopontin (Opn) is one of the co-factors involved in cell adhesion and invasion during the implanta- tion process. Several reports have shown Opn expression changes in diabetic condition in several tissues. In addition, an increased incidence of spontaneous abortion is reported in diabetic women. We, therefore, designed a study to eval- uate the effects of diabetes on Opn expression at implantation time after treatment with metformin and pioglitazone. Materials and Methods In this interventional and experimental study, 28 rats were randomly divided into four groups, namely control, diabetic, pioglitazone-treated diabetic rats and metformin-treated diabetic rats. Streptozo- tocin (STZ) and nicotinamide (NA) were used to induce type 2 diabetes (T2D). During the implantation window, the endometrium was removed and the expression of Opn was analysed by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Results Opn expression was significantly higher (30.70 fold-changes) in the diabetic group in comparison with the control group (P=0.04). Furthermore, the expression of Opn was significantly lower in the diabetic group treated with pioglitazone when compared with the diabetic group (P=0.04). Conclusion According to the high Opn expression and the possibility of increased adhesion of endometrial epithelial cells, the invasion of blastocyst may be affected and thus reduced. As pioglitazone significantly reversed the upregula- tion of Opn in diabetic rats, it may be considered as a therapeutic compound for treating T2D.

Neurodegenerative diseases have now become a major challenge, especially in aged societies. Most of the traditional strategies used for treatment of these diseases are untargeted and have little efficiency. Developments in stem cell investigations have given much attention to cell therapy as an alternative concept in the regeneration of neural tissues. Dental pulp stem cells (DPSCs) can be readily obtained by noninvasive procedures and have been shown to possess properties similar to well-known mesenchymal stem cells. Furthermore, based on their neural crest origin, DPSCs are considered to have a good potential to differentiate into neural cells. Zfp521 is a transcription factor that regulates expression of many genes, including ones involved in the neural differentiation process. Therefor based on neural crest origin of the cell and high expression of neural progenitor markers, we speculate that sole overexpression of Zfp521 protein can facilitate differentiation of dental stem cells to neural cells and researchers may find these cells suitable for therapeutic treatment of neurodegenerative diseases.

Beta-thalassemia is one of the most common autosomal recessive disorders in the world population resulting from over 200 different mutations of HBB gene. Beta-thalassemias are caused by point mutations or, more rarely, deletions in the HBB gene leading to reduced (beta+) or absent (beta0) synthesis of the beta chains of hemoglobin (Hb). High-resolution melting of polymerase chain reaction (PCR) products can detect heterozygous and most homozygous mutations without electrophoretic or chromatographic separations. In the current study, blood samples collected from 20 individuals carrying minor thalassemia were genotyped using HRM technique. The genotype of each sample had been previously determined via the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)/ amplification-refractory mutation system (ARMS) or sequencing method. This study aimed to determine the specificity and sensitivity of HRM method in the diagnosis of carriers of FSC 36-37 (-T) mutation from carriers who do not have this mutation. DNA extraction from peripheral blood was performed and HRM method was used to genotype samples. The results were analyzed according to the normalized and difference plot. High-resolution melting analysis could correctly identify all carriers of FSC 36-37 (-T) from who did not have this mutation. In summary, HRM is a technique associated with high sensitivity and specificity. Therefore, HRM is an appealing technique for the identification of FSC 36-37 (-T) mutation.

BackgroundMetabolic syndrome (MetS) is a complex and multifactorial disorder characterized by insulin resistance, dyslipidaemia, hyperglycemia, abdominal obesity, and elevated blood pressure. The apolipoprotein A5 (APOA5) gene variants have been reported to correlate with two major components of MetS, including low levels of high density lipoprotein cholesterol (HDL-C) and high levels of triglyceride. In the present study, we explored the associations between five single nucleotide polymorphisms (SNPs) of APOA5 gene and the MetS risk.MethodsIn a case-control design, 120 Iranian children and adolescents with/without MetS were genotyped by polymerase chain reaction-sequencing for these SNPs. Then, we investigated the association of SNPs, individually or in haplotype constructs, with MetS risk.ResultsThe rs34089864 variant and H1 haplotype (harboring the two major alleles of rs619054 and rs34089864) were associated with HDL-C levels. However, there was no significant association between different haplotypes/individual SNPs and MetS risk.ConclusionThese results presented no association of APOA5 3’UTR SNPs with MetS. Further studies, including other polymorphisms, are required to investigate the involvement of APOA5 gene in the genetic susceptibility to MetS in the pediatric age group.

Objective(s): As the second cause of cancer death, gastric cancer (GC) is one of the eminent dilemmas all over the world, therefore investigating the molecular mechanisms involved in this cancer is pivotal. Unrestricted proliferation is one of the characteristics of cancerous cells, which is due to deficiency in cell regulatory systems. Long non-coding RNAs (lncRNAs) have emerged as critical regulators of the epigenome. lncRNA extra coding CEBPA (ecCEBPA) is involved in DNA methylation. This lncRNA reduces CEBPA promoter methylation by interacting with DNA methyltransferase 1. lncRNA UCA1 (urothelial carcinoma-associated 1) elevates cell proliferation through the PI3K/Akt signaling pathway which has a critical role in cell growth and apoptosis. The aim of this study was to examine the expression of ecCEBPA and UCA1 genes in GC tissues as well as their clinical significance. Materials and Methods: Total RNA extraction, cDNA synthesis, and quantitative real-time PCR were performed for cells and 80 paired GC tissues. Furthermore, clinical relevance of UCA1 expression was investigated in TCGA cohort data. Results: Our results showed ecCEBPA and UCA1 over-expression in GC tissues. Furthermore, lncRNAs associations with clinicopathological features were demonstrated both in the current and TCGA cohort. Kaplan-Meier analysis indicated that patients with higher UCA1 expression had a worse overall survival in the case of pancreatic and lung adenocarcinomas but not other solid cancer types including GC. Conclusion: These data demonstrate UCA1 and ecCEBPA involvement in GC and suggest that these lncRNAs might be useful as diagnostic/ prognostic biomarkers in cancer.

Introduction: The polymorphic enzyme cytochrome P450 2D6 (CYP2D6) catalyzes a major step in the bioactivation of tamoxifen. Genotyping of clinically relevant CYP2D6 alleles and subsequent dose adjustment is a promising approach to individualize breast cancer therapy. The aim of this study was to investigate the relationship between the plasma levels of tamoxifen and its metabolites and different CYP2D6 genotypes under standard (20 mg/day) and dose-adjusted therapy (Registration ID in Iranian Registry of Clinical Trials: IRCT2015082323734N1).Materials and Methods: Using TaqMan® assays common alleles of CYP2D6 (∗1, ∗2, ∗4, ∗5, ∗6, ∗10, ∗17, and ∗41) and gene duplication were identified in 134 breast cancer patients. Based on CYP2D6 genotypes patients with an activity score 1 (n = 15) and 0–0.5 (n = 2) were treated with tamoxifen adjusted dosage of 30 and 40 mg/day, respectively. The concentration of tamoxifen and its metabolites before and after 4 and 8 months of dose adjustment were measured using LC-MS/MS technology.Results: At baseline, (Z)-endoxifen plasma concentrations (33 ± 15.5, 28.1 ± 14, 26.6 ± 23.4, 14.3 ± 8.6, and 10.7 ± 5.5 nmol/l for EM/EM, EM/IM, EM/PM, IM/IM and PM/PM, respectively) and the metabolic ratio (Z)-Endoxifen/N-desmethyltamoxifen (0.0558 ± 0.02, 0.0396 ± 0.0111, 0.0332 ± 0.0222, 0.0149 ± 0.0026, and 0.0169 ± 0.0177 for EM/EM, EM/IM, EM/PM, IM/IM, and PM/PM, respectively) correlated with CYP2D6 genotype (Kruskal–Wallis p = 0.013 and p < 0.0001, respectively). Dose escalation to 30 and 40 mg/day in patients with a CYP2D6 activity score of 1 (n = 15) and 0–0.5 (n = 2) resulted in a significant increase in (Z)-endoxifen plasma levels (22.17 ± 24.42, 34.43 ± 26.54, and 35.77 ± 28.89 nmol/l at baseline, after 4 and 8 months, respectively, Friedman p = 0.0388) along with the plasma concentrations of tamoxifen and its other metabolites. No severe side effects were recorded during dose escalation.Conclusion: For the first time, we show the feasibility of dose escalation of tamoxifen in breast cancer patients with compromised CYP2D6 activity and Iranian ethnic background to increase the plasma concentrations of (Z)-endoxifen.

Background:The zinc-finger X linked (ZFX) gene encodes a transcription factor that acts as a regulator of self-renewal of stem cells. Due to the role of ZFX in cell growth, understanding ZFX protein-protein interactions helps to clarify its proper biological functions in signaling pathways. The aim of this study is to define ZFX protein-protein interactions and the role of ZFX in cell growth. Materials and Methods: The PIPs output includes three interacting proteins with ZFX: eukaryotic translation initiation factor 3 subunit I(EIF3I), eukaryotic translation initiation factor 3 subunit G(EIF3G) and protein nuclear pore and COPII coat complex component homolog isoform 3 (SEC13L1). Results: As a cargo and transmembrane protein interacting with Sec13,eIF3I and eIF3G, ZFX mediates cargo sorting in COPII vesicles at ER exit sites. While traveling to cis-Golgi, eIF3I is phosphorylated by the mechanistic target of rapamycin (mTOR). Proteins transport by COPI vesicles to the nucleusouter site layer containing SEC13 via the contribution of microtubules. EIF3G and eIF3I interact with coatomer protein complex subunit beta 2 (COPB2) that helps to enclose ZFX in COPI vesicle. ZFX and eIF3G enter nucleolus where activation of transcription from pre rDNA genes occurs. Conclusion:We proposed a model in which ZFX is involved in cell growth by promoting the transcription of rDNA genes.

مقدمه: سرطان‌های بدخیم کولورکتال، چه در صورت تکی و چه در صورت خانوادگی، اگر با افزایش میزان ناپایداری در میکروستلایت همراه باشند، پیش‌آگهی بهتری دارند، راهکار متفاوتی برای جراحی در آن‌ها اتخاذ می‌شود و پاسخ متفاوتی نسبت به داروی شیمی‌درمانی 5FU (Fluorouracil) خواهند داد. بنا بر این، امروزه برای بیماران مبتلا به سرطان کولورکتال ارثی و غیر ارثی، آزمایش سنجش وضعیت ناپایداری میکروستلایت انجام می‌شود. هدف این مطالعه، سنجش کارایی نشانگر تک نوکلئوتیدی MT1XT20 در جمعیت بیماران مبتلا به سرطان کولون ارثی غیر پولیپی (HNPCC یا Hereditary nonpolyposis colorectal cancer) کشور ایران بود. علاوه بر نشانگر پیش‌گفته، از دو روش بررسی وضعیت میکروستلایت با پنل 5 نشانگری و استاندارد Promega و نیز آزمایش ایمنوهیستوشیمیایی استفاده شد. روش‌ها: در این مطالعه، نشانگر تک نوکلئوتیدی MT1XT20 و کیت استاندارد Promega از طریق روش PCR (Polymerase chain reaction) بر روی 20 خانواده‌ی مبتلا به HNPCC ایرانی مورد سنجش قرار گرفت و آزمایش ایمنوهیستوشیمیایی نیز برای بررسی وضعیت 4 پروتئین مهم دخیل در مسیر ترمیم ناجور جفت شدگی مورد استفاده قرار گرفت. یافته‌ها: در روش سنجش ناپایداری میکروستلایت از طریق پنل Promega و نشانگر تک نوکلئوتیدی MT1XT20 به ترتیب 8 (40 درصد) و 5 نمونه (25 درصد) از 20 نمونه وضعیت ناپایداری نشان دادند. در روش ایمنوهیستوشیمیایی نیز، 7 نمونه از 20 نمونه (35 درصد) وضعیت کمبود پروتئین مسیر ترمیم ناجور جفت شدگی را نشان دادند. در میان نشانگرهای مورد استفاده در کیت Promega، نشانگر BAT26 ناپایدارترین نشانگر بود و در هر 8 نمونه، Microsatellite instability-high (MSI-H) تشخیص داده شده توسط کیت Promega ناپایداری نشان داد. نشانگرهای NR21 و NR24 در 7 نمونه (5/87 درصد) ناپایداری نشان دادند و نشانگرهای BAT25 و MONO27 به ترتیب با نشان دادن ناپایداری در 6 (0/75 درصد) و 5 نمونه (5/62 درصد)، پایدارترین نشانگرها بودند. نتیجه‌گیری: MT1XT20 در جمعیت ایتالیایی، به عنوان نشانگر تک نوکلئوتیدی مورد اعتماد ارزیابی شده است، اما این مسأله در رابطه با جمعیت ایرانی صدق نمی‌کند. در عوض، BAT26 با نشان دادن ناپایداری در تمام نمونه‌های MSI-H ارزیابی شده توسط کیت Promega، می‌تواند به عنوان نشانگری موثق و قابل مقایسه با پنل 5 نشانگری Promega در جمعیت ایرانی در نظر گرفته شود.

Background: Beta-thalassemia is one of the most common autosomal recessive disorders in the world population caused by more than 200 different mutations in the beta-globin chain. It is clinically classified as minor, intermediate and major. Beta-thalassemia is the most common monogenic disease in the Mediterranean countries, Middle East, Indian Subcontinent, and Southeast Asia and one of the widespread hereditary disorders in Iran. Among different

Metabolic syndrome (MetS) is a prevalent disorder in children and it is described as a group of risk factors including obesity, increased fasting blood sugar, increased triglyceride, high blood pressure and decreased high-density lipoprotein (HDL) levels. This multifactorial disorder has been considered as one of the leading causes of mortality throughout the world and has been considered as a major risk factor for type 2 diabetes mellitus and atherosclerotic cardiovascular diseases. Recent studies have shown that the syndrome is highly prevalent among overweight children and adolescents. The etiology of the metabolic syndrome is complex, and is determined by the interaction between genetic and environmental factors and their influence on the development of obesity, insulin resistance and various inflammatory processes. Recent findings on Iranian children and adolescents show the prevalent of 23%. In this review, we looked at the definition, prevalence etiology and the main determinant genes in the development of this disorder.

Background: Integral membrane proteins involve in a variety of valuable cellular processes including signal transduction, transport and cell-to-cell communication. Thus, due to their importance, are targeted to treatment of diseases or disorders. To achieve this goal, it is necessary to obtain adequate membrane proteins. A variety of expression systems have been used to overexpress these proteins but mammalian cells can produce membrane proteins structurally close to natural form of them and are favourable for this purpose. TOSO/FAIM3 (Fas Apoptosis Inhibitory Molecule), highly conserved membrane protein, playing important role in cell survival, immune surveillance, and homeostasis and in this study was overexpressed on HEK-293T membrane. Methods: Eukaryotic expression vector pEZ-M67-TOSO was transformed into Escherichia coli top 10F’ strain to multiply. Then, the plasmid was extracted and digested with restriction enzyme Eco31I to make a linear plasmid. Subsequently, HEK-293T cells were transfected with linear plasmid and treated with hygromycin to select TOSO expressing stable cells. After three weeks, resistant colons were expanded, Chromosomal DNA and total RNA were extracted and the presence of TOSO-cDNA in HEK-293T genome and the expression level of TOSO were evaluated using polymerase chain reaction (PCR) and Real-time PCR methods, respectively. Findings: The integration of TOSO expressing cassette in HEK-293T genome was confirmed. HEK-293 cells expressed 1700 mRNA TOSO molecule/cell which was in the range of protein with abundant expression. Conclusion: Mammalian cells represent membrane proteins in desirable form versus other expression systems. For this reason, we used HEK293T cells to overexpress transmembrane TOSO protein. In this investigation, we successfully constructed the HEK293 cell line with stable TOSO overexpression, which will facilitate further researches to characterize biophysical and biochemical structure and function of TOSO and pharmaceutical biological researches about it.

Objective(s):Previous studies demonstrate that changes in pre-mRNA splicing play a significant role in human disease development. Furthermore, many cancer-associated genes are regulated by alternative splicing. There are mounting evidences that splice variants which express predominantly in tumors, have clear diagnostic value and may provide potential drug targets. Located on the X chromosome, ZFX gene functions as a transcription regulator for self-renewal of stem cells. This gene has 5 splice variants that encode 3 isoforms. In the present study, we evaluated the clinicopathological relevance of the expression of ZFX isoform 3/variant 5 gene in gastric carcinoma. Materials and Methods: A total of 60 tumoral and non-tumoral gastric specimens were evaluated for ZFX isoform 3/variant 5 gene expression using quantitative real-time PCR. Results: Our results showed that the expression of ZFX isoform 3/variant 5 transcript was heterogeneous in gastric specimens. We further showed that there was a positive correlation between the variant expression and tumor size, but not with other clinicopathological features of gastric tumors. Conclusion: This report shows that the expression of ZFX isoform 3/variant 5 transcript was heterogeneous in gastric specimens. Furthermore, there was no significant association between ZFX isoform 3/variant 5 expression and most of clinicopathological features of gastric tumors except for a positive correlation with tumor size. The elucidation of the precise molecular mechanisms governed by the ZFX isoforms/variants needs further investigation.

Background: Considering the high prevalence of congenital hypothyroidism (CH) in Isfahan and its different etiologies in comparison with other countries, the high rate of parental consanguinity, and the role of NIS gene in permanent CH due to dyshormonogenesis, the aim of this study was to investigate the G395R mutation of the NIS gene in patients with permanent CH due to dyshormonogenesis Methods: In this case-control study, patients diagnosed with permanent CH due to dyshormonogenesis during CH screening program were selected. Venous blood sample was obtained to determine the G395R mutations of NIS gene using polymerase chain reaction (PCR) sequencing method. Results: In this study, 35 CH patients with permanent CH due to dyshormonogenesis and 35 neonates with normal screening results as a control group were studied. We did not find any changes of the mentioned mutation of NIS gene in the patients′ group. Conclusion: Considering the findings of the current study, it seems that further studies with larger sample size and with consideration of other gene mutations such as pendrin and thyroglobulin are needed for more accurate conclusion.

• Background: Utilizing an intact RNA is essential while performing molecular genetics methods like microarray and quantitative Real-time polymerase chain reaction (RT-PCR). Using RNA with poor quality may affect the results of downstream manipulations. Our aim in the current study was to compare and optimize various procedures for RNA extraction by using different kits and DNase treatment to find a standard method for RNA extraction from human frozen tissues. • Methods: Using Cinnagen RNX plus solution, Qiagen RNeasy Mini Kit and Qiagen RNeasy Plus Mini Kit, total RNA was extracted from human frozen tissues comprising stomach (#40), glioma (#5) and breast (#15). For genomic DNA elimination, extracted RNA was treated with DNase enzyme. After RNA extraction, its quantity and quality was assessed by spectrophotometry and gel electrophoresis. After cDNA synthesis, RT-PCR technique by specific primers designed for TBP gene was performed and results were assessed by agarose gel electrophoresis. • Finding: Extracted RNA by Cinnagen RNX plus solution did not show good results in RT-PCR after DNase treatment. Extracted RNAs by Qiagen RNeasy Mini Kit and Qiagen RNeasy Plus Mini Kit were of high quality for RT-PCR reaction because of using DNase treatment during the extraction procedure and an additional genomic DNA elimination column respectively. • Conclusion: Combining Qiagen RNeasy Mini Kit, Qiazol solution and DNase treatment during the extraction procedure is the most suitable method for RNA extraction from human frozen tissues and the extracted RNA can be used with assurance in downstream molecular techniques.

Background: Diabetes, a major metabolic disorder, seems to affect the fertility rates of women in various ways. Due to the uncertainty of the effects of diabetes along with superovulation treatment on the infertility, we investigate the effects of ovulation induction treatment as therapeutic approach on the expression of leukemia inhibitory factor (LIF) and vascular endothelial growth factor A (VEGFA) as two main factors which are involved in the implantation in the streptozotocin (STZ)-induced type 1 diabetic rats. Materials and Methods: Type 1 diabetes was induced by injections of STZ in Wistar rats. The animals were kept in diabetic conditions for 4 weeks, while some were treated with insulin for treatment. After treatment, the ovulation was induced by human menopausal gonadotropin (hMG) and human chorionic gonadotropin (hCG). The rats were then sacrificed and the expression of LIF and VEGFA was checked by immunohistochemistry staining method, and the relative expression of LIF and VEGFA was measured by quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting methods. Results: It was observed that diabetes and insulin treatment for diabetes altered the expression of Lif and VEGFA in both mRNA and protein levels. However, superovulation treatment seems to ameliorate this alternation for both factors. Conclusion: According to our results, diabetes and insulin therapy could alter the expression of Lif and VEGFA genes and proteins that are effective in endometrial receptivity and implantation process. It seems in diabetic cases, the effect of hCG and hMG therapy by itself could regulate the level of expression and presence of these two genes and proteins.

Cytochrome P450 2D6 (CYP2D6) is an important drug-metabolizing enzyme involved in the pharmacokinetic metabolism of drugs. CYP2D6 gene is highly polymorphic, and the combination of its different alleles yields different phenotypes including extensive metabolizer (EM), intermediate metabolizer (IM), poor metabolizer (PM), and ultrarapid metabolizer (UM). Genotyping of the important alleles for this gene in different ethnicities is of particular importance for assessing the efficacy of various drugs. In this study, we reviewed the CYP2D6 allele and phenotype frequencies predicted from the genotypes of CYP2D6 in the Middle East area. Regardless of different ethnicities, the CYP2D6*41 allele frequency was shown to be higher than that of other reduced functional alleles. In addition, CYP2D6*4 was the most frequent nonfunctional allele in all studied populations in the Middle East. Taken together, our findings illustrated that the frequencies of PM or IM alleles and different genotypes harboring these alleles are relatively high in the Middle Eastern countries. Therefore, the study of CYP2D6 alleles for each patient to detect those that are at risk is of great importance to prevent adverse drug reactions through individualization therapy.

Background: Cancer is the second cause of death after cardiovascular diseases worldwide. Tumor metastasis is the main cause of death in patients with cancer; therefore, unraveling the molecular mechanisms involved in metastasis is critical. Epithelial-mesenchymal transition (EMT) is believed to promote tumor metastasis. Based on the critical roles of long noncoding RNA-ATB (lncRNA-ATB) and SETD8 genes in cancer pathogenesis and EMT, in this study, we aimed to assess expression profile and clinicopathological relevance of these two genes in human gastric cancer. Materials and Methods: Quantitative real-time polymerase chain reaction was performed to assess these gene expressions in gastric cancer tissues and various cell lines. The associations between these gene expressions and clinicopathological characteristics were also analyzed. Results: Insignificant downregulation of lncRNA-ATB and significant upregulation of SETD8 in cancerous versus noncancerous gastric tissues were observed. Among different examined cell lines, all displayed both genes expression. Except for a significant inverse correlation between the expression levels of lncRNA-ATB and depth of invasion (T) and a direct association between SETD8 levels and advanced tumor grades, no significant association was found with other clinicopathological characteristics. Conclusion: lncRNA-ATB and SETD8 genes may play a critical role in gastric cancer progression and may serve as potential diagnostic/prognostic biomarkers in cancer patients.

Background: Familial hypercholesterolemia (FH) is a disorder that is inherited by autosomal dominant pattern. The main cause of FH disease is the occurrence of mutations in low-density lipoprotein receptor (LDLR) gene sequence, as well as apolipoprotein B and proprotein convertase subtilisin/kexin type 9 genes, located in the next ranks, respectively. Materials and Methods: Forty-five unrelated Iranian patients with FH were screened using a high-resolution melting (HRM) method for exon 9 along with intron/exon boundaries of LDLR gene. Samples with shift in resultant HRM curves were compared to normal ones, sequenced, and analyzed. Results: Our findings revealed a missense mutation c. 1246C>T and a known variant IVS9-30C>T (rs1003723) that was recognized in 71% of the patients (22%: homozygous and 49%: heterozygous genotypes). In silico analysis, predicted the pathological effect of the c. 1246C>T mutation in LDLR protein structure, but IVS9-30C>T variant had no predicted effect on splice site and branch point function. Conclusion: FH is a hereditary type of hypercholesterolemia that leads to premature cardiovascular disease and atherosclerosis, and early diagnosis is needed. We detected a rare missense mutation (1246C>T) and a common single nucleotide polymorphism (SNP) in the Iranian population. These reports could help in the genetic diagnosis and counseling of FH patients.

Background: Patients affected by gliomas have a poor prognosis. Astrocytoma is a subtype of glioma. Identification of biomarkers could be an effective way to an early diagnosis of tumor or to distinguish more aggressive tumors that need more intensive therapy. In this study, we investigated whether the expression of miR-362 was increased or decreased in patients with different grades of astrocytoma. Materials and Methods: miR-362 expression was compared in 25 patients with astrocytoma with that of 4 normal nonneoplastic brain tissues. Results: In all tumor tissues, the expression of miR-362 was significantly decreased relative to its expression in normal brain tissues. However, there was no significant difference between miR-362 expressions in high and low grades of astrocytoma. Conclusions: In conclusion, miR-362 showed a down-regulation pattern in astrocytoma tissues that was different from the pattern obtained from previously published microarray studies.

Background: Metabolic syndrome (MetS) is a common disorder which is a constellation of clinical features including abdominal obesity, increased level of serum triglycerides (TGs) and decrease of serum high-density lipoprotein-cholesterol (HDL-C), elevated blood pressure, and glucose intolerance. The apolipoprotein A5 (APOA5) is involved in lipid metabolism, influencing the level of plasma TG and HDL-C. In the present study, we aimed to investigate the associations between four INDEL variants of APOA5 gene and the MetS risk. Materials and Methods: In this case–control study, we genotyped 116 Iranian children and adolescents with/without MetS by using Sanger sequencing method for these INDELs. Then, we explored the association of INDELs with MetS risk and their clinical components by logistic regression and one-way analysis of variance analyses. Results: We identified a novel insertion polymorphism, c. *282–283 insAG/c. *282–283 insG variant, which appears among case and control groups. rs72525532 showed a significant difference for TG levels between various genotype groups. In addition, there were significant associations between newly identified single-nucleotide polymorphism (SNP) and rs72525532 with MetS risk. Conclusions: These results show that rs72525532 and the newly identified SNP may influence the susceptibility of the individuals to MetS.

Background: Metabolic syndrome (MetS) is a prevalent disorder in pediatric age groups, described by a combination of genetic and environmental factors. Sterol regulatory element-binding transcription factor 1 (SREBF-1) induces the expression of a family of genes involved in fatty acid synthesis. Moreover, dysregulation of miR-33b, which is located within the intron 17 of the SREBF-1 gene, disrupts fatty acid oxidation and insulin signaling, thus leading to MetS. The aim of the present study was to investigate the association between SREBF-1 rs8066560 polymorphism and MetS in Iranian children and adolescents. Methods: This study includes 100 MetS and 100 normal individuals aged 9-19 years. Anthropological and biochemical indexes were measured. The -1099G > A polymorphism was genotyped by TaqMan real-time polymerase chain reaction. Results: Significant differences were observed in anthropometric measurements and lipid profiles between MetS and normal children. There were no differences in the genotype frequencies or allele distribution for -1099G > A polymorphism between MetS and control groups. High-density lipoprotein cholesterol levels were significantly higher in the MetS GG group than in the A allele carrier group. The genotype AA controls had significantly increased cholesterol and low-density lipoprotein cholesterol levels than AG genotypes. By logistic regression using different genetic models, no significant association was observed between SREBF-1 rs8066560 polymorphism and the risk of MetS. Conclusions: We conclude that the -1099G > A variant on SREBF-1 gene associated with serum lipid profiles, however, it may not be a major risk factor for the MetS in Iranian children and adolescents.

Background: Congenital hypothyroidism (CH) due to the thyroid dyshormonogenesis is more prevalent in Iran in comparison to other countries. Sodium iodide symporter (NIS) is one of the plasma membrane glycoproteins that is located on the basolateral side of thyroid follicular cells and mediates active I− trapping into these cells. Playing a prominent role in thyroid hormone synthesis, NIS gene mutations can be a cause of permanent CH with the etiology of dyshormonogenesis. The aim of this study was to investigate the occurrence of T354P mutation of the NIS gene, in a group of children affected with permanent CH in Isfahan. Materials and Methods: Thirty-five patients with the etiology of dyshormonogenesis, and 35 healthy children, collected between 2002 and 2011 in Isfahan Endocrine and Metabolism Research Center, were examined for T354P mutation of the NIS gene by direct polymerase chain reaction-sequencing method. Results: No T354P mutation was detected in any of the studied children. Conclusions: More subjects with confirmed iodide transport defects should be screened for detecting the frequency of different reported NIS gene mutations in our population.

Background: Gastric cancer is the second and fourth most common cancer in Iranian men and women, respectively, but it is the first leading cause of cancer deaths in Iran. Most Iranian patients with gastric cancer are diagnosed at an advanced stage of disease when the conventional treatments have no effect on improving the survival. So, early gastric cancer detection is of high priority in order to decrease its high mortality rate in Iran. HOTAIR is a long non-coding RNA which its overexpression has been documented in different types of human cancer and can be considered as a potential cancer biomarker. The aim of this study was to evaluate the clinicopathological relevance of the expression of HOTAIR gene in gastric carcinoma. Materials and Methods: A total of 60 tumoral and non-tumoral gastric specimens were evaluated for HOTAIR gene expression using quantitative real-time PCR. Results: The expression of HOTAIR was markedly increased in gastric cancer tissues compared with adjacent non-tumoral tissues. We further showed that there was a positive significant correlation between the HOTAIR gene expression, TNM staging, perineural invasion, and distant metastasis, but not with other clinicopathological features of gastric tumors. Conclusions: These results suggest that HOTAIR expression is modulated during gastric cancer progression and therefore may participate in molecular processes relevant to malignant transformation and metastasis in gastric carcinoma.