Evaluating changes in the expression of BCL-2 gene, lncRNA SRA, and miR-361-3p in unexplained recurrent pregnancy loss.

Unexplained recurrent pregnancy loss (RPL) composed almost half of all diagnosed miscarriage cases. As the apoptosis pathway is involved in the pregnancy process the present investigation aimed to assess the differential expression of the BCL-2 gene, SRA lncRNA, miR-361-3p in unexplained RPL patients. In this study, RNA was isolated from 50 blood samples of people with a history of RPL, and 50 blood samples of people with healthy fertility. After cDNA synthesis from these samples, alterations in the expression levels of the above-mentioned genes were examined by Real-Time PCR. Our results showed that the expression of BCL-2 and lncRNA SRA was significantly higher in the blood samples of RPL patients than in controls, while the expression of miR-361-3p was significantly downregulated. Besides, there were significant correlations between the changes in the expression of lncRNA SRA and miR-361-3p with BCL-2, in positive and negative directions, respectively. Also, miR-361-3p presented as a good diagnostic marker with the highest AUC value to discriminate between RPL and the healthy control subjects. These results proposed that ncRNAs may have a significant role in the regulation of apoptosis relates genes expression in RPL.

Photobiomodulation and Stem Cell on Repair of Osteoporotic Bones.

Objective: This study examined the use of photobiomodulation (PBM) plus adipose-derived stem cells (ASCs) to enhance the osteogenic properties of demineralized bone matrix (DBM) scaffold in a critical size femoral defect (CSFD) of ovariectomy-induced osteoporotic (OVX) rats. Background: PBM could be used as a unique strategy to enhance the osteogenic potential of DBMs seeded with ASCs. Materials and methods: The OVX rats with a CSFD were divided into six groups: (1) Control (C); (2) DBM scaffold alone (S); (3) S+PBM; (4) S+alendronate; (5) S+ASC; (6) S+PBM+ASC. Stereological analysis, real-time polymerase chain reaction (RT-PCR), and cone-beam computed tomography (CBCT) were performed after euthanization at 4 and 8 weeks postimplantation surgery. Results: In the 8th week, Groups 4 and 6 showed a greatly high new trabecular bone volume than the scaffold group (all, p = 0.009). The CBCT data demonstrated that the CSFD was significantly smaller in the two, three, and six groups relative to the control group (p = 0.01, p = 0.000, and p = 0.000, respectively). RT-PCR revealed that Groups 3 and 6 had higher messenger RNA levels of osteocalcin (OC) and osteoprotegerin (OPG) compared with the control group (p = 0.05). Group 6 had significantly lower expression of receptor activator of nuclear factor-κB ligand (RANKL) compared with the control group (p = 0.02). Conclusions: The combination of DBM plus PBM plus ASC, as well as DBM plus PBM significantly improved the healing of CSFD in OVX rats, and affected the expression of OPG, OC, and RANKL genes.

Aberrant expression of BAX, MEG3, and miR-214-3P genes in recurrent pregnancy loss.

AIMS: Recurrent pregnancy loss (RPL), with unknown causes, is one of the most common challenges facing pregnancy. Apoptotic signaling pathways are involved in the normal and abnormal pregnancy process. Despite the evidence pointing toward the aberrant expression of apoptotic and apoptotic-related genes in pregnancy complications, the involvement of these genes in RPL remains to be elucidated. This study aimed to investigate the expression levels of BAX, MEG3, and miR-214-3p (as a microRNA), and their associations in an Iranian population. MATERIALS AND METHODS: Following the extraction of RNA from blood samples of RPL patients and controls, quantitative expression levels of BAX, MEG3, and miR-214-3p genes were analyzed by real-time RT-PCR. RESULTS: The findings showed that the expression levels of BAX and miRNA-214-3p were significantly higher in the blood samples of RPL patients than in control samples. In contrast, the expression of MEG3 was significantly down-regulated in women RPL. Furthermore, altered expressions of MEG3 and miRNA-214-3p are associated with their target gene BAX, where the BAX expression is positively and negatively correlated with the expressions of has-miR-214-3P and MEG3, respectively. ROC curve evaluation demonstrated the highest specificity and diagnostic value for miR-214-3p expression in distinguishing RPL samples from the healthy controls. CONCLUSIONS: These data indicated that the aberrant expression of BAX, MEG3, miRNA-214-3p genes in RPL patients could provide new insights into the biological characteristics and related pathways of differentially expressed genes, which could help as potential diagnostic biomarkers and a better understanding of the molecular mechanisms of RPL.

Hsa-miR-27a-3p and epidermal growth factor receptor expression analysis in glioblastoma FFPE samples.

AIM: Glioblastoma multiforme (GBM) is the most invasive type of glial tumors. MicroRNAs as the small, noncoding RNAs have been revealed that play an important role in tumorigenic processes. So, they may be used as potential biomarkers for detection and prognosis of cancers at the early stages. In addition, they can be applied as therapeutic targets. In the present study, the expression levels of hsa-miR-27a-3p and EGFR were investigated in GBM. METHODS: Real-time RT-PCR was applied to evaluate hsa-miR-27a-3p and EGFR expressions in the formalin-fixed paraffin-embedded (FFPE) tissue samples obtained from 50 GBM and 50 normal people. RESULTS: The expression level of hsa-miR-27a-3p and EGFR was significantly different between cases and controls. Positive association was found between gene expressions and immunohistochemistry markers, such as Ki67 and glial fibrillary acidic protein, except for IDH1 status. CONCLUSION: We showed the association of hsa-miR-27a-3p and EGFR with GBM and it can be concluded that they have a promising potential to use as primary cancer biomarkers.

Downregulation of Cancer-Associated lncRNAs in Peripheral Blood of Multiple Sclerosis Patients.

Recent studies have shown contribution of long non-coding RNAs (lncRNAs) in the pathogenesis of immune-related disorders including multiple sclerosis (MS). Based on the role of these transcripts in the regulation of immune response, peripheral levels of lncRNAs can reflect the level of immune activation. In the present study, we quantified expression of four lncRNAs namely SPRY4-IT1, HOXA-AS2, LINC-ROR, and MEG3 in venous blood of MS patients and controls using quantitative real-time PCR method. Relative expressions of SPRY4-IT1, HOXA-AS2, LINC-ROR, and MEG3 were significantly lower in female MS patients compared with female healthy subjects. For MEG3, this pattern of expression was also observed in male subjects. However, for other lncRNAs, no significant difference was detected between male patients and male controls. Expression of HOXA-AS2 was correlated with progression index (r = 0.36, P < 0.001). Besides, there was a significant correlation between expression of this lncRNA and expression of LINC-ROR in MS patients (r = 0.44, P < 0.0001). There was no other correlation between expression of lncRNAs and clinical data in MS patients. In control group, expressions of none of lncRNAs were correlated with age of persons. Notably, significant correlations were demonstrated between expression levels of all lncRNAs in healthy subjects with r values ranging from 0.23 to 0.42. The current investigation shows dysregulation of lncRNAs in MS patients in a sex-specific manner and warrants further studies to unravel the clinical and therapeutic implications of such dysregulation.

Down-regulation of RORA gene expression in the blood of multiple sclerosis patients.

Multiple sclerosis (MS) is an autoimmune disease characterized by recurrent episodes of demyelination and loss of oligodendrocytes. The demyelination process is caused by various subsets of CD4+ T cells with a Th1 and Th17 phenotype. The retinoid acid-related orphan receptor A (RORA) is expressed in Th17 cells and promote Th17 differentiation. In this study, we compared the expression level of RORA gene in the blood of 50 relapsing-remitting MS (RRMS) patients who were treated with IFN-ß and 50 healthy controls by TaqMan Quantitative Real-Time PCR.We found that RORA expression was significantly down-regulated in MS patients compared with controls (P= 0.006). However, there was no significant correlation between RORA gene expression and Kurtzke Expanded Disability Status Scale (EDSS). Our findings suggest a possible contribution of IFN-ß in the downregulation of RORA. In addition, RORA downregulation may be a potential indicator of positive response to interferon beta treatment of multiple sclerosis patients.

STAT5a and STAT6 gene expression levels in multiple sclerosis patients.

Multiple sclerosis (MS) is a complex inflammatory, autoimmune disease of the central nervous system (CNS). The disease pathogenesis is not well defined yet. Cytokines have an important role in inflammation as characteristic feature of the disease. Janus kinase/signal transducers and activators of transcriptions (JAK/STAT) family promote cytokine-mediated cell activation. Failure in the JAK/STAT signaling pathway is associated with the pathological outcome in MS. In this study, we compared the expression levels of STAT5a and STAT6 genes in the blood of 50 relapsing-remitting MS (RR-MS) patients and 50 healthy controls by Taqman Quantitative Real-Time PCR in patients and healthy control group. We found that STAT5a expression was significantly down-regulated (p = .049), whereas STAT6 gene expression was significantly up-regulated (p = .046) in MS patients compared with controls. Moreover, there was significant correlation between the STAT6 gene expression and Kurtzke Expanded Disability Status Scale (EDSS) criterion. However, no significant correlation was demonstrated between the expression of STAT5a gene and clinical findings. Furthermore, there was not significant correlation between expression levels of STAT5a and STAT6 genes. Our findings suggest that STAT5a and STAT6 dysregulation may have a critical role in modification of immune responses leading to imbalance between Th2- and Th1-related cytokines. However, the mechanisms underlying it still remain to be elucidated. Future studies are needed to explore the role of STAT5a and STAT6 as prognostic biomarkers in research, design of experimental therapies or clinical settings of the MS.