Journal of

Background and Aim: MicroRNAs (miRNA) are small noncoding RNA molecules that transcribed by RNA polymerase II. After biogenesis, these molecules act by incorporation into the RNA-induced silencing complex (RISC). MiRNAs are involved in multiple physiological and pathological processes such as proliferation, differentiation, apoptosis and cancer.
Recently several studies reported down regulation of mir-150 during erythropoesis. Since hemoglobin expression is valuable indicator of erythroid differentiation we evaluated the mir-150 downregulation effect on alpha chain expression by Quantitative RT-PCR.

Materials and Methods: K562cells were grown in RPMI1640 in standard condition. K562 cells were transfected by microRNA 150 Inhibitor using transfection kit .Mir-150 downregulation was confirmed by miRNA Real time PCR, followed by Q-RT-PCR to investigate the alpha chain expression changes.

Results: By relative QRT-PCR the alpha chain expression was increased 10 folds in comparison to untransfected and scramble cells. Furthermore, the differences were statistically significant (P<0.05)

Conclusion: Elevation of alpha chain expression in our study showed that mir-150 downregulation has a crucial role in erythroid differentiation and can introduce as a novel marker in alpha thalassemia. Further researches to find out the detail mechanism and miRNAs genes target could improve our knowledge about miRNAs potential in management of diseases and their applications in gene therapy and regenerative medicine.

Background and Aim: Embryonic stem cells are identified with two unique characteristics. First, they can be maintained and expanded as pure populations of undifferentiated cells, a characteristic which is known as self renewal aspect of embryonic stem cells. Second, these cells can give rise to all body cell types. In the current study, we used a feeder-free condition to differentiate mouse embryonic stem cells into lymphoid lineage by IL-7 and FLT-3 ligand.

Materials and Methods: Mouse embryonic stem cells cultured on mouse embryonic fibroblasts were separated from the feeder layer. Then, embryoid bodies were formed from mouse embryonic stem cells. Following that, differentiation was performed by FLT-3 ligand and IL-7. In order to demonstrate the differentiation into lymphoid lineages, the expression of CD25, CD19 and CD3 was assessed by RT-PCR technique on days 7 and 14.

Results: After 14 days of differentiation into lymphoid lineages by defined factors, RT-PCR results showed the expression of CD25 and CD19 markers.

Conclusion: In all previous studies, mouse embryonic stem cells were differentiated into lymphoid lineage by OP9 stromal feeder cells. In this study, a feeder-free condition was used to differentiate mouse embryonic stem cells into lymphoid lineage. It is hoped that the present study can lead to new insights in cell therapy of lymphoid deficiency disorders.

 Background and Aim: Understanding the molecular mechanisms involved in the increased levels of HbF inducing drugs should be advised for effective induction. The aim of this study was to investigate the molecular effects of the drugs thalidomide and sodium butyrate considered as HbF inducer agents.

 Materials and Methods: In this experimental study, CD133+ cord blood stem cells carrying mutations of heterozygous β-thalassemia were isolated and differentiated into erythroid lineage. In order to evaluate the expression of the erythroid markers, CD71 and CD235a, was analysed. For this purpose, the RNA extracted from erythroid precursors at days 6 and 12 of erythroid differentiation and cDNA synthesized, and then the expression of these genes was performed by quantitative Real-time PCR technique.

 Results: The results of this study showed the significant effect of thalidomide on erythroid proliferation as compared to sodium butyrate and control group (P<0.05). Also, thalidomide significantly increased CD71expression and decreased CD235a expression as compared to sodium butyrate and control groups (P<0.05).

 Conclusion : Thalidomide may play its role on HbF induction by increasing the proliferation of early erythroid precursors.

Background and Aim: Obesity is now considered as one of the main risk factors of certain known diseases such as cardio-vascular diseases, non- insulin-dependent diabetes, and common cancers. Moreover, the increase of white fat tissue is known as a main factor in the obesity process, in terms of physiology and pathology. Therefore, the understanding of adipocytes differentiation processes is crucial.

Materials and Methods: In this study, mesenchymal stem cells (MSCs) were isolated from human bone marrow by ficol-gradient, and then, their surface markers were confirmed by flow cytometry. Osteoblastic and adipocytes differentiation were done by dexamethasone protocol and confirmed by staining. Then qualitative and quantitative expressions of PPARgamma (PPAR-γ) gene as an important transcription factor in the differentiation of fat were studied by RT-PCR and REAL TIME PCR before and after differentiation into adipocytes. For statistical analysis, paired t-test was performed, using pfaffl and graph pad software.

Results: PPAR-gamma gene expression showed a significant increase after differentiation of human bone marrow mesenchymal stem cells into adipocytes (p<0/05).

Conclusion: According to the results, the PPAR-γ gene acts as one of the important factors in the differentiation of MSCs into adipocytes. In brief, the inhibition of this gene's expression to prevent obesity is suggested as an idea for treatment in the future.

Background and Aim: Vaginal delivery and cesarean are two different methods of delivery. These two different conditions could affect the number and types of predominant white blood cells of mothers. This study is aim to assess and determine the number and types of white blood cells of mothers according to vaginal and elective cesarean.

Materials and Methods: That induced among pregnant women referred to Malayer hospital in Iran between 2011-2012. 30 pregnant women with vaginal, and 30 women with elective cesarean were selected as sample size simultaneously. The blood specimen of pregnant women was analysed by cell counter, sysmex model KX-21 before and after delivery. The data was analysed by SPSS Software using descriptive tests, analysis of variance, and multivariance analysis along with P< 0.05 as a significant level.

Results: The results showed that there was a significant correlation between increase of white blood cells count of two groups. The average of segmented Neutrophils of vaginal delivery group was significant increased in comparison with selective cesarean women.

Conclusion: Stress and physical pressure in vaginal delivery could affect the increase of white blood cells in comparison with cesarean delivery. Although, there is no evidence to approve the impression increased of predominant white blood cells on delivered mother and newborn at birth.

Background and Aim: Different regulation processes have an effect on osteoblastic differentiation of mesenchymal stem cells (MSCs), and among them Wnt signaling pathway is particularly desirable. In Wnt signaling pathway, Adenomatous Polyposis Coli (APC) bind to β-catenin and induce its degradation, thereby acting as a negative regulator of canonical Wnt pathway. In this study, gene expression and DNA methylation of APC gene during osteoblastic differentiation were determined.

Materials and Methods: In this experimental study, after the isolation of MSCs, the induction of osteoblastic differentiation was done. To confirm osteoblastic differentiation, alizarin red staining together with the expression of Alkaline Phosphatase (ALP) and osteocalcin as specific osteoblastic markers was performed. APC gene methylation status by MSP (Methylation Specific PCR) and gene expression status of APC gene using Real-Time PCR technique during different times were evaluated.

Results: The results of alizarin red staining and the expression of ALP and osteocalcin confirmed osteoblastic differentiation. In addition, the results showed a significant decrease in the expression of APC gene on the 7^th day of osteoblastic differentiation (P<0.05). Also, the results revealed hypermethylation status of APC gene promoter during osteoblastic differentiation.

Conclusion: It seems that the decreased expression APC gene will play an important role in Wnt signaling pathway regulation in different stages during osteoblastic differentiation of bone marrow-derived MSC. Also, according to the results, APC gene promoter methylation will play an important role in controlling gene expression.