فصلنامه خون
سال دوازدهم شماره 3 (پیاپی 49، پاییز 1394)

To organize blood donors, promote blood donation, predict the future, and detect possible intervening factors, we must monitor and evaluate the trend of blood donation. In this cross-sectional study, all blood donors during the 5-year period from 2009 to 2013 in Kermanshah entered the study. The data analyzed using Excel, statistical package of Stata (version 12), and independent T-test were used to compare the means in different groups; Poisson regression was used to investigate the significant trend during the time. During 2009 to 2013 (5 years), 295925 potential donors referred to blood centers in Kermanshah province; 249420 (84.2%) were eligible for blood donation out of whom 216563 (87.8%) were men. The trend of blood donation during the years of study showed to be downward among females but upward among males. Moreover, among blood donors 117080 (46.9%) were regular donors whose number was growing. Findings show that most blood donors in the West of Iran are regular donors with an ascending trend. Given the fact that regular donors are safer than first-time donors for infections such as HBV, HCV and HIV, it can be concluded that over time the quality and safety of blood donations would be at a better position.

Micro RNAs are a class of small non-coding RNAs which have been recently shown to play a crucial role in major cellular processes such as development and differentiation through post-transcriptional regulation. The correlation of microRNA expression levels with the pathogenesis of some hematologic malignancies including acute myeloid leukemia is already known. The aim of this study was to investigate the expression of mir-150 and mir-155 in chronic myelogenous leukemia patients. miRNA expression level changes can be used for diagnosis, treatment and estimation of Minimal Residual Disease (MRD). In this descriptive study, the expression of miR-150 and miR-155 in 25 newly diagnosed CML patients in chronic phase and 25 healthy subjects as the control all in Payvand Medical Laboratory were examined by Stem-loop RT-PCR and Real Time PCR techniques. For data analysis, Pearson correlation analysis was used. miR-150 and miR-155 were down-regulated in CML patients rather than in healthy subjects (0.512 and 0.2552, respectively). There was no correlation between the laboratory findings and the miR expression level. But miR-155 was inversely associated with age (p ≤ 0.01). In conclusion, the decreasing expression of miR-150 and miR-155 in chronic myelogenous leukemia is indicative of their induced expression in suppressing proliferation of cells in this disease. Other studies are also required to elucidate the exact role of microRNA by identifying their target genes.

HLA system plays a key role in the cancer cells’escape from the immune surveillance. AML is a cancer of the myeloid lines of blood cells, characterized by the rapid growth of abnormal white blood cells in the bone marrow. Several studies worldwide have investigated the association of AML with alleles of HLA. This study was performed to assess the association between alleles of HLA-A, -B,-DRB1 with AML patients in Kerman province of Iran. In the present case control study, alleles of HLA-A, -B, -DRB1 were molecularly typed using PCR-SSP technique in 33 AML patients and 270 healthy individuals unrelated with patients in province of Kerman, Iran. Finally, the data analysis was performed using c 2 and SPSS 19. In this study, allelic frequency rates of HLA-A*11 in patient and control groups were 28.8% (19 cases) and 17% (92 cases), respectively; it indicates a significantly positive association between the presence of this allele and AML illness (p = 0.02, OR = 1.97 and CI (95%) = 1.10-3.51). The presence of HLA-A* 11 in AML patients may be a predisposing factor for this disease to develop.

Cord blood CD133+ stem cells are highly proliferative primitive hematopoietic progenitors. These cells can also differentiate to other cells especially neural cells. The PAX6 gene belongs to a family of genes that play a critical role in the formation of tissues and organs during embryonic development. The PAX6 protein is thought to activate genes involved in the formation of the eyes, brain and spinal cord (central nervous system), and the pancreas. The purpose of this study was to assess PAX6 (5a) gene transfer in cord blood CD133+ stem cells and evaluate its effect on differentiation. In this experimental study, cord blood stem cells were collected and mononuclear cells isolated by ficoll; then, CD133+ cells were obtained using the CD133 MicroBead Kit in combination with the autoMACS Separator. These cells were cultured in Stem Span media. Next, HEK293T packaging cells were co-transfected with PLEX-MCS, PsPAX2, and pMD2G by calcium phosphate method. Lentiviral vectors were collected and concentrated. The appropriate amount of viruses was used to infect CD133+ cells. The successful transduced cells were selected by puromycin resistance. After two weeks, the expression of Rhodopsin, CHX10, Thy1, Nestin, and PAX6 proteins was assessed by immunocytochemisrty method. Two weeks after infection, the expression of Rhodopsin, CHX10, Thy1, Nestin, and PAX6 proteins were detected in treatment cells and the expression of Rhodopsin and Nestin in control cells. The results showed CD133+ cells differentiated into progenitor neural like cells and retinal neural like cells including ganglion like cells.

Due to the unique criteria, mesenchymal stem cells (MSCs) are the ideal cells for cell therapy and gene therapy. However, the low survival of MSCs after transplantation has limited their application. This study aimed to evaluate the expression of cytoprotective genes including NQO1, TXNRD1, HO-1, GCLC following the overexpression of Nrf2 in MSCs. In this experimental study, umbilical cord-derived MSCs were cultured and recombinant vectors containing Nrf2 and empty vectors were transfected into MSCs using FuGENE HD. After exposure of the cells to stress, RNA extraction and cDNA generation were performed. Using Primer3 software, specific primers were designed for Nrf2, NQO1، TXNRD1، HO-1 and GCLC genes and the expression of these mentioned genes was evaluated by RT-PCR. The results were quantified and analyzed statistically utilizing Image J software and ANOVA. The expression of Nrf2 was up-regulated in MSCs after transfection (p< 0. 01). Overexpression of TXNRD1 and GCLC was observed in transfected cells (p <0. 05 and p <0. 01); however, the expression of NQO1 and HO-1 did not change in the transfected group in comparison to the control (p > 0. 05). Overexpression of Nrf2 resulted in the overexpression of TXNRD1 and GCLC in MSCs and might be explained by the fact that a part of the known Nrf2 cytoprotective mechanisms is controlled by the expression of these genes.

Umbilical cord blood, as a source of mesenchymal stromal cells, has many advantages compared to other sources. This study aimed to provide a new method for the in vitro neural differentiation of human umbilical cord blood derived mesenchymal stromal cells (MSCs hUCB). In this experimental study, MSCs hUCBs were isolated and characterized by morphologic, adipogenic, and osteogenic differentiation and immunophenotypical analysis. Then, neural induction of MSCs hUCB was performed by using RA, bFGF, NGF, EGF, AsA, IBMX, and neurobasal medium. Then, the relative expression of neural-specific genes was investigated by quantitative real-time PCR assays, REST 2009, and SPSS 11.5 software. Our results showed the osteocytic and adipocytic differentiation capacity and fibroblast-like morphology of MSCs hUCB. Flow cytometry analysis of MSCs hUCB revealed that the cells are positive for CD105 (78%), CD73 (78%), and negative for CD45 (2%), HLA-DR (2.5%). The cells showed the remarkable transition from fibroblast-like morphology to nueral progenitor cells. The results showed that the expression of GFAP, MBP, nestin, MAP-2 genes after neural induction significantly increased in comparison with that of the control as measured by quantitative real-time PCR assays (p < 0.05). Treatment of MSCs hUCB with a set of growth factors and chemical materials induces neural differentiation and increases the efficiency of cell-based therapy for neurodegenerative diseases in the future. Although, the functionality of neural progenitor cells must be carefully assessed in animal models prior to use in clinical application.

Cord blood CD133+ cells are able to maintain long-term hematopoiesis and to differentiate to different hematopoietic lineages. CXCR4 overexpression is involved in successful transplantation of hematopoietic stem cells in the bone marrow. Platelet micro particles (PMP) contain CXCR4 markers and are able to transfer them into hematopoietic stem cells. Therefore, considering the importance of CD133+ cells as primitive HSCs, the effect of platelet micro particles on the expression levels of CXCR4 and CD34 markers in these cells was examined. this experimental study, cord blood CD133+ cells were isolated by MACS. Isolated cells were cultured into three groups one as control without adding PMP and the other two groups with PMP with concentrations of 5 and 10 μg/mL. The cells were cultured for five days in stem span medium. Expression of CXCR4 surface marker was analyzed by flow cytometery; the total cell number was counted by hemocytometer and colony forming units (CFU) were measured by colony assay. Fold increase of CD34+ and CXCR4+ cells in the control group and in the presence of 5 and 10 µ g/mL of PMP were 52.6 ± 4.16, 64.2 ± 6.91, and 67.2 ± 6.76, respectively (p= 0.047). CXCR4+ cell percentage in the presence of 10 µ g/mL PMP compared to control cells (63.8 ± 6.4) was 72.8 ± 4 (p= 0.049). Exposure of CD133+ cells isolated from cord blood to PMP with 10 μg/mL concentration increased the expression of CXCR4 surface marker significantly.

The tissue donation is a complex phenomenon which involves the physical, psychological and social dimensions. People's perception of this phenomenon depends on cultural norms, social factors, and individual knowledge of tissue donation. This study is conducted to explore the experiences of physicians about the voluntary donation of tissues. In 2014, this qualitative phenomenological study was done on 14 physicians in a hospital in Uremia; the participants had a tissue card. To gather the data, the semi-structured interview was used. All recordings and handwritten information were analyzed with phenomenology of VAN Manen approach. The five major themes were extracted from the data analysis: the experience of physicians caring for the patients in need of tissue donation, altruism, faith, patterning, and the obstacle of family disapproval. They are the factors explaining the experiences of physicians with tissue donation. Physicians have had the unique experiences about voluntary organ donation. Therefore, it is necessary for all medical care staff to understand the complexities of the physicians’ experiences in preparatory stages for tissue donation.

Bone marrow microenviroment contains cellular and acellular compartments. Cellular compartment contains hematopoetic stem cells, mesenchymal stem cells and some other kinds of stromal cells while acellular compartment includes scaffold proteins known as an extra cellular matrix. Hematopoietic stem cells resided in the bone marrow interact with a bone marrow microenvironment called the stem cell niche. Hematopoietic stem cells (HSCs) are able to produce the variety of blood cells and bone marrow microenvironment plays a supportive role for the cells in this process. Recent studies over in-vitro models demonstrating the essential role of stromal cells in hematopoiesis reflected the view that cell-cell contact in the marrow microenvironment is critical for hematopoietic stem cells normal function and differentiation. Direct cell-cell contacts and cytokines secreted by mesenchymal stem cells play a critical role in the coculture of hematopoetic stem cells and mesenchymal stem cells and the determination of the fate of hematopoetic stem cells. Several studies demonstrate the effect of mesenchymal stem cells on self renewal, expansion, proliferation and differentiation of hematopoetic stem cells in vitro that lead to different and contradictory results. In this paper, we will investigate the effect of mesenchymal stem cells on differentiation of hematopoietic stem cells in vitro. The data of the present article were obtained through the review of many papers published on the effect of stem cells. The review of different studies has shown the necessity to survey the effect of mesenchymal stem cells on differentiation of different cell lines as a laboratory model. Direct cell-cell contact between mesenchymal stem cells and hematopoietic stem cells and the signaling pathways activated by this interaction enable proliferation and expansion of HSCs in the undifferentiated state.