Cell-free seminal mRNA of DDX4 and TNP1 Genes as Potential Biomarkers of the Presence of Sperm in the Testicular Tissue

Introduction: Non-obstructive azoospermia (NOA) is one of the reasons for infertility in men, and different factors including genetic factors are involved in its development. Since taking biopsies of the testicular tissue for assisted reproductive technologies (ARTs) is invasive and time-consuming, and the testicular tissue is heterogeneous, introducing a biomarker for predicting the possibility of the presence of sperm in the testicle can increase the ART efficiency. Accordingly, Cell-free seminal mRNA (CFs-mRNA), which is found in many fluids of the body including the seminal fluid of NOA individuals, can be employed as a biomarker for this purpose. Materials and methods: This study was conducted on 15 men suffering NOA, candidates for testicular sperm extraction (TESE), along with 15 healthy men as control. The testicular tissue of 10 patients was examined using hematoxylin and eosin staining and then classified according to Johnsen scoring. RNA was extracted from the cell-free plasma of semen samples and cDNA was synthesized. The Expression level of TNP1 and DDX4 genes was investigated using real-time polymerase chain reaction (PCR). Results: The expression of CFS-mRNA of the DDX4 gene was observed in only one sample of NOA individuals (10%), showing a score of 8. Further, the expression of CFS-mRNA of the TNP1 gene was observed only in two samples (20%) of NOA patients whose scores were 3 and 8. Conclusion: Insufficiency or lack of expression of CFS-mRNA of TNP1 and DDX4 genes may be helpful in predicting the absence of sperm in the testicular tissue of NOA patients in terms of sperm retrieval for ART. Yet, further studies with more specific and sensitive techniques are required to achieve a more solid and precise conclusion

Increased expression of stemness genes Rex-1, Oct-4, Nanog, and Sox-2 in women with ovarian endometriosis versus normal endometrium: A case-control study

Background: Endometriosis is a common, chronic inflammatory disease which is defined as an overgrowth of endometrial tissue outside the uterine cavity. The etiology of this disease is complex and multifactorial but there is a strong evidence that supports the presence of endometrial stem cells and their possible involvement in endometriosis. Objective: In this study, we analyzed the mRNA expression of REX-1 stemness gene and reconsidered three other stemness genes SOX-2, NANOG, OCT-4 in women with endometriosis compared to normal endometrium. Materials and Methods: Ten ectopic and ten eutopic tissue samples along with 23 normal endometrium specimens were recruited in this study. The expression levels of OCT-4, NANOG, SOX-2, and REX-1 genes were evaluated by the quantitative real-time polymerase chain reaction. Results: The transcription levels of OCT-4, NANOG, and SOX-2 mRNA were significantly increased in ectopic lesions compared with eutopic and control group (p = 0.041, p = 0.035, p = 0.048), although the REX-1 mRNA increase was not significant between endometriosis and control groups. Also, there were differences in the expression level of these genes in normal endometrium during the menstrual cycles (p = 0.031, p = 0.047, p = 0.031). Conclusion: Based on our data, we confirm the dynamic role of stemness genes in proliferation and growth of normal endometrium during the menstrual cycle and conclude that differential expression n levels of these genes may contribute to the pathophysiology of endometriosis. Key words: Endometriosis, Stemness genes

Basic and clinical genetic studies on male infertility in Iran during 2000-2016: A review

The male factor contributes to 50% of infertility. The cause of male infertility is idiopathic and could be congenital or acquired. Among different factors which are involved in idiopathic male infertility, genetic factors are the most prevalent causes of the disease. Considering, the high prevalence of male infertility in Iran and the importance of genetic factors in the accession of it, in this article we reviewed the various studies which have been published during the last 17 yr on the genetic basis of male infertility in Iran. To do this, the PubMed and Scientific information database (SID) were regarded for the most relevant papers published in the last 17 yr referring to the genetics of male factor infertility using the keywords „„genetics‟‟, “cytogenetic”, „„male infertility”, and “Iranian population”. Literatures showed that among the Iranian infertile men Yq microdeletion and chromosomal aberrations are two main factors that intervene in the genetics of male infertility. Also, protamine deficiency (especially P2) is shown to have an influence on fertilization rate and pregnancy outcomes. The highest rate of sperm DNA damages has been found among the asthenospermia patients. In several papers, the relation between other important factors such as single gene mutations and polymorphisms with male infertility has also been reported. Recognition of the genetic factors that influence the fertility of Iranian men will shed light on the creation of guidelines for the diagnosis, consultation, and treatment of the patients.&quot

Molecular investigation of c-MYC oncogene amplification in diabetic patients of Karaj County, Iran: Investigating c-MYC Amplification in Diabetics

Introduction: There is growing evidence revealing that genetic factors could be involved in the etiology of insulin resistance and diabetes. Recent studies now suggest that c-MYC oncogene amplification in beta-cells can cause downregulation of insulin gene expression and leading to diabetes. The present study was carried out to examine gene amplification level of c-MYC gene in healthy individuals compared to those with diabetes. Materials and Methods : This case control study consisted of 70 subjects (34 diabetic patients and 36 healthy controls). The genomic DNA was extracted from blood samples using standard phenol-chloroform method, and Differential Quantitative PCR (DQ PCR) was accomplished. Subsequently, the mean Band Intensity Ratio (BIR) of c-MYC to γ-IFN was applied to determine the amplified products' molecular band intensity. The Real-time PCR was used to reconfirm the obtained results. Results: The mean BIR of c-MYC to γ-IFN was 1.29 in diabetics, whereas in healthy individuals, the mean BIR was equal to 1.16.The findings indicate that the mean BIR in diabetics was almost 1.1 times higher than healthy controls; however, it was not statically different (P-value= .168). Conclusions: Since no significant difference was found in terms of BIR of c-MYC to γ-IFN  in case and control groups, it can be concluded that c-MYC may not have a role to trigger the disease in this limited Iranian population. Further studies and larger statistical populations are needed to confirm our findings. This study showed that the DQ PCR technique could be reliable to screen gene amplification in large populations