Background: Coronary artery disease (CAD) refers to stenosis or obstruction in a part or all of a coronary artery due to atherosclerosis or clotting. This study aimed to evaluate the possible efficacy of the serum microRNA-197 (miR-197) as an indicator of diagnosis in patients with CAD. Methods: In this study, 100 patients with CAD who had angiography and vascular transplantation were selected and evaluated. The expression level of miR-197 was determined via the real-time RT-PCR technique and the SYBR Green method. For the analysis of the miRNA expression level and the significance of the patient sample, the t test was used. Additionally, the Pearson correlation coefficient test was utilized to determine the relationship between the expression levels of miRNAs and CAD severity. Results: A positive correlation was observed between miR-197 expression and CAD severity. The average expression of 0. 78 in the control sample was increased to 2. 76 according to the severity of involvement in the patient. In other words, the relative expression of miR-197 in the CAD + group was significantly increased compared with the control group (P<0. 004). Conclusions: It appears that miR-197 can be considered an indicator of coronary endothelial cell function, and it is possible to use it as a biomarker for the prognosis, control, or treatment of CAD.

Objective: A class of small non-coding RNAs (microRNAs) has been recently identified as critical molecules involved in a wide spectrum of normal and pathological processes. MicroRNAs (miRNAs) are negative regulators of gene expression and function as cytoplasmic gene silencers by suppressing translation of targeted mRNAs. They have a tumor specific profile which is variable in different tumor grades and stages. In this study, the expression of miR-21 (an oncomir upregulated in a variety of tumors) and miR-302 (an embryonic stem cells-specific miRNA, expressed in some cancer stem cells) was evaluated in formalinfixed paraffin-embedded (FFPE) samples of esophagus cancers. FFPE archival tissue samples are a valuable source of materials for determining the retrospective prognostic value of miRNA profiling in human cancers.Materials and Methods: Tumor and non-tumor (adjacent, apparently-normal tissue of the same patients) samples of esophagus squamous cell carcinoma (SCC) were collected from the archival collection of Shiraz university of medical sciences. The tumor parts of the whole blocks were carefully punched off to avoid contamination with non-tumor parts of the blocks. Deparaffinization procedure as well as proteinase K digestion process was optimized using different strategies and solutions. Then, total RNA extraction was performed using Trizol and RNXsolutions. cDNA synthesis and real-time RT-PCR were performed using Exiqon uniRT cDNA synthesis kit and SYBR green master mix. MiRCURY LNA™ microRNA detection probes were employed to localize subcellular distribution of miR-21, and miR-302 by means of in-situ hybridization (ISH). The ISH procedure was optimized by using different Proteinase K solutions and hybridization temperatures on an internal control (U6 snRNA). Results: We detected miR-21 and miR-302 expression in several stem and tumor cell lines, including KYSE30 cell line of esophagus. We also optimized a practical technique for total RNA extraction from FFPE samples and detected miR-21 and miR-302 overexpression in tumor samples compared to their non-tumor counterparts. We also detected the nuclear localization of U6 snRNA, as a control non-coding RNA, using ISH technique and by means of LNA probes. Conclusion: We were able to extract and amplify miRNAs from FFPE tumor samples. The obtained data are of great clinical importance and have potential usefulness to introduce new tumor markers capable of predicting the malignant behavior of different types of tumors.

Hemoglobinopathy is an important genetic problem affecting millions of world population. The new alternative treatment is the use of gene therapy. The repair based on small nuclear RNAs (SnRNAs) is a widely discussed method. It is proved and validated for many hemoglobinopathies with underlying beta globin gene defects [1]. Many recent publications indicate the feasibility of using U7.623 gene therapy for several beta hemoglonopathies such as hemoglobin (Hb) C [2] and S disorders [3]. However, this approach has never been tested for the Hb J Iran (beta 77 His----Asp), which is a problematic hemoglobinopathy in Middle East [4]. For a brief assessment, the author uses the previously published in silico method by Wiwanitkit for assessment of recovery of function and biological process of hemoglobin Iran after application of U7.623 gene therapy [2-3]. The validity and reliability of the used method is approved [2-3]. Based on the gene ontology study, the complete recovery cannot be determined. The question is why this technique is not applicable for treatment of hemoglobin J Iran. Basically, the rationale of using U7.62 is that it contains a sequence antisense to a region between the aberrant splice sites, therefore it reduces the incorrect splicing of pre-mRNA and leads to increased levels of the correctly spliced b-globin mRNA and protein. Therefore, it is justified to use U7 in certain mutated Hbs (such as mutations at 654, 705 or 745 in intron 2 of the Hb) because these mutations can activate aberrant 3' and 5' splice sites within the intron, prevent correct splicing of beta-globin pre-mRNA and result in inhibition of beta-globin synthesis [5]. Hence, this is the reason that using U7 snRNA in hemoglobin J Iran where His77 is substituted by Asp is not applicable. Although this article did not report on any novel but negative finding, the report in this work is different from the previous reports by the authors in the serial study of gene therapy for hemoglobin disorders that highlighted the efficacy of U7.623 gene therapy in several beta hemoglobinopathies [2-3]. It is interesting that U7.623 gene therapy cannot be applicable for treatment of hemoglobin J Iran and it is required to find and test other alternative gene therapeutic approaches.

Tuberculosis (TB) remains a major threat to human health. Understanding the strategies mycobacteria take to overcome immune defense is important in order to control the infection. Micro (mi)RNAs are master regulators of most pathways in the human body. Infection with mycobacterium impacts upon the host metabolic pathways as they are subverted to obtain the nutrition for intracellular TB survival. In this study, we aimed to investigate the effect of Bacillus Calmette-Gué rin (BCG) infection on the expression of three miRNAs (miR-1224,-484 and-425), which are important in infection and in the regulation of metabolic pathways. Peripheral blood monocyte-derived macrophage (MDM) cultures were prepared and infected with BCG at a multiplicity of infection (MOI)=10 or left uninfected as a control. 72h post-infection, RNA was extracted from the cultured cells and cDNA synthesis and real-time PCR performed. Expression levels miRNAs were normalized to the levels of U6 snRNA (Rnu6) using the 2– Δ Δ Ct method. Infection with BCG resulted in a highly significant increase in miR-1224 expression (24. 4± 3. 8-fold induction) in human MDMs. The induction of miR-484 (1. 8± 0. 3-fold increase) and of miR-425 (1. 2± 0. 2-fold increase) was less increased compared to miR-1224. Mycobacterium tolerates a hostile microenvironment by escaping from lysosomal degradation and providing a lipid-rich niche by trigger with and re-pattering host metabolism. This study highlighted the potential roles of miRNAs in host responses upon mycobacterium infection.

Objective: microRNAs (miRNAs) are a new class of non-coding RNAs involved in regulating various biological processes including proliferation, differentiation, and apoptosis, among others. Alterations in miRNA expression are reported in several human cancers, which suggest their potential roles in tumor initiation and progression. Members of the miR-302 cluster are highly expressed in embryonic stem cells (ESCs), where they regulate cell self-renewal and pluripotency. Based on the cancer stem cell (CSC) hypothesis, mis-expression of such genes might contribute to tumorigenicity. This study aims to find a potential link between the expression level of human/homo sapiens miR-302b (has-miR302b) and tumor/grade state of gastric tissues.Materials and Methods: A matched based case-control study was conducted that included tumor and matched marginal non-tumor surgical specimens from 34 patients diagnosed with gastric adenocarcinoma. Realtime reverse transcriptionpolymerase chain reaction (RT-PCR) assays were performed with specific LNATM primers and SYBR Green master mix. The human embryonic carcinoma cell line, NTERA2 (NT2) and a human gastric adenocarcinoma cell line, AGS, were used as controls. The ability of miR-302b to discriminate tumor from non-tumor gastric samples was evaluated using the area under the receiver operating characteristic (ROC) curve.Results: According to our data, miR-302b expression (normalized to that of the U6 snRNA housekeeping gene) in the pluripotent cell line NT2 was more than 500 times greater than that of the AGS cell line. The level of expression was even lower in tumor and nontumor gastric tissue samples. The data further revealed a down-regulation of miR-302b in gastric tumor samples (p=0.001), particularly in high-grade adenocarcinoma (p=0.009). However, ROC analysis data demonstrated a low sensitivity and specificity of miR-302b expression to discriminate between the tumor and non-tumor state of the samples (AUC=0.63).Conclusion: Despite the upregulation of some hESCspecific genes in tumors, our data revealed a downregulation of miR-302b in high-grade tumors. This data suggested a potential tumor-suppressor role for miR- 302b in tumorigenesis of gastric tissue.