JOURNAL OF CELL & TISSUE
Year:2016 | Volume:7 | Issue:1|Papers Count:10

Aim: The aim of this study was to generate recombinant lentiviruses carrying Nurr1 to express it in human cells.Material and methods: The IRES-EGFP fragment was isolated from the pIRES2-EGFP vector using restriction enzymes BglII/ NotI and made blunt-ended using Klenow. The transfer vector PNLEGFP/ CMV/WPREdU3 was digested with NheI/XhoI and made blunt-ended. Finally, the isolated IRES-EGFP fragment was inserted into this lentivirus vector to generate lentivirus construct (I). The human Nurr1 gene was then isolated from the PCMX-NOT vector using BamHI and XhoI and inserted into construct (I) pre-digested with BamHI and SalI. At this step lentivirus construct (II) as our final transfer construct was generated. In order to generate recombinant lentiviruses, we then transfected the HEK-293T cell line with transfer vector plus packaging and envelope vectors. Cell medium full of virus particles was collected and passed through Amicon filters to produce a concentrated virus stock. The stock was ultimately used for transduction of fresh HEK-293T cells.EGFP expression was shown under fluorescent microscope and Nurr1 expression was analyzed using RT-PCR.Results: Enzymatic tests confirmed the correct cloning of the hNurr1gene into the lentivirus backbone. Observation by fluorescent microscopy showed EGFP expression post-transfection and post-transduction. RT-PCR demonstrated Nurr1 expression at both stages.Conclusion: In this study, lentiviruses carrying the human Nurr1 gene were produced and used for transduction of human cell line HEK-293T. The transduced cells successfully expressed the Nurr1 gene.

Aim: The aim of this study was to evaluate the anti-diabetic effects of Tragopogon graminifolius hydroethanolic leaf's extract (TGE) on blood glucose and insulin level in diabetes male rats.Material and methods: In this experimental study 42 Wistar male rats were randomly divided in 6 groups: control, diabetic (streptozotocin60 mg/kg, i.p), experimental diabetic groups (treatment with TGE, 200, 400 and 800 mg/kg, i.p) and diabetic treatment with metformin (500mg/kg, gavaged) for 10 days. The blood glucose was examined daily by glucometer. At the end of examination blood samples and pancreatic tissue were collected for insulin evaluation and histological study. All data are expressed as mean±SEM and statistical significance was considered at p<0.05.Results: Treatment with the 800 mg/kg TGE decreased significantly blood glucose in diabetic rats compared with control group (P<0.05). In addition, administration of 400 and 800 mg/kg of TGE could significantly decreased plasma glucose levels compared with metformin group. The insulin blood levels increased significantly in treated groups compared with diabetic group (P<0.05).Conclusion: The TGE has flavonoid and phenolic compositions and its prescription might be decrease blood glucose and increase blood insulin levels in diabetic rats.

Aim: The aim of this study was to investigate the effect of different concentrations of Aluminum oxide nano-particles on seed germination, root length, and the amounts of photosynthetic pigments, total protein content, and changes in activity of some antioxidant enzymes and carbohydrates content in Phaseolus vulgaris.Material and methods: Experiment was performed under greenhouse conditions and completely randomized designed with four replications. Plants were exposed to different concentrations (0.01, 0.5 and 1 g/L) of nano-Aluminum oxide and the physiological and biochemical characteristics of treated plants were compared with control ones.Results: The results showed that treatment by Aluminium oxide nano-particles had a positive impact on the seed germination, root length, total chlorophyll and chlorophyll b content and also sugar content. Decrease in the content of chlorophyll a, protein and catalase activity was observed in the treated plants in comparison to control. Aluminium oxide nano-particle didn’t have a significant effect on seed germination speed and peroxidase activity.Conclusion: Based on the results of this study, nano-particles are able to have a positive effect on some developmental and physiological characteristics of Phaseolus vulgaris L. This indicated that applied nano-particles are not toxic in the used concentrations. Decreasing in characteristics (chlorophyll a, total protein and catalase activity) was observed that could be related to plant resistance.

Aim: The aim of this study was to describe the possible effects of vitrification on maturation rate and ultrastructural morphology of in-vitromatured human oocytes.Material and Methods: A total of 292 immature Germinal Vesicle (GV) and Metaphase I (MI) oocytes obtained from infertile patients were allocated into two groups: (i) GV oocytes (n=145), (ii) MI oocytes (n=147). Oocytes were first vitrified and then matured in-vitro. Supernumerary fresh invivo matured oocytes (n=10) were used as control. in-vitromaturation media was Ham’s F10 supplemented with FSH+LH and human follicular fluid. After 36h of incubation, the oocytes were investigated for nuclear maturation and ultrastructural changes.Results: The rate of survival and degeneration was significantly higher in Metaphase I than GV group.But, Oocyte maturation rates were not significant between groups (P<0.06). In addition, the rate of oocyte arrest was significantly higher in MI oocytes in comparisson with GV oocytes.Ultrastructurally; a drastic reduction was observed in amount of cortical granules at the cortex of vitrified-warmed GV and MI oocytes, In addition, Transmission Electron Microscopy (TEM) revealed vacuoles and small mitochondria-Smooth Endoplasmic Reticulum aggregates in the ooplasm of oocytes.conclusion: Freeze/thawing procedures are associated with ultrastructural alterations in specific oocyte microdomains, presumably related to the reduced competence of cryopreserved oocytes to maturation.

Aim: The present study aims to investigate if artesunate exerts its anti-proliferative activity by increasing antioxidant enzymes activity in MCF-7 human breast cancer cell line. Determining the oxidant effect of artesunate may elucidate a possible alternative mechanism for its cytotoxicity.Material and methods: For evaluating cytotoxic activity of artesunate, MCF-7 cells were treated with different concentrations (0, 1, 5, 10, 25, 50, 75, 100 and 200 mg/ml) of artesunate and subjected to MTT assay after 24 hours. Also, the enzymatic activities of superoxide dismutase (SOD), and peroxidase (POD) were measured in MCF-7 cells treated with selected doses of artesunate (0, 10, 25, 50 and 100 mg/ml) after 24h. In addition, the cell culture supernatants were used to assess the amount of nitric oxide (NO) production using the Griess method.Results: Artesunate inhibited the growth of MCF-7 cells, dose-dependently and also significantly increased the activity of antioxidant enzymes: superoxide dismutase (SOD), and peroxidase (POD).Furthermore, it suppressed NO production, dose-dependently.Conclusion: To conclude, it seems that artesunate exert its cytotoxic activity by increasing the activity of antioxidant enzymes and through inhibition of NO production in MCF-7 cells. The increased activities of antioxidant enzymes in the treated cells could alter the antioxidant defense system, potentially contributing towards the anti-proliferative effect.

Aim: This study aimed to evaluate the histomorphometric and immunohistochemical parameters of the repair of femoral bone defect using bone marrow stromal cells on gelatin– chitosan membrane in adult Albino Wistar rats.Materials and Methods: In this experimental study, sixty male Albino wistar adult rats were equally divided into five groups as follows: Control group that received no treatment after bone defect. Sham group that after bone defect, the culture medium was injected locally at the site of defect. Gelatinchitosan group that membrane was used into bone defect. Cell group that nonautolog BMSCs were injected locally into defect. Cell-GC group that cell transplantation with chitosan - gelatin membrane were used into the bone defect.Results: The mean area of trabeculae in groups of membranes and cells significantly increased when compared to the control group. The mean number of osteocytes and cells in the bone defect in cell group significantly increased when compared to the control group. No significant difference were found in chitosan - gelatin and sham groups compared to control group but the mean number of osteocytes significantly decreased in BMSCs with gelatin-chitosan scaffold group compared to control group (P<0.001) Conclusion: BMSC transplantation and gelatin-chitosan scaffold are effective in repair of bone defect.However, the use of BMSCs with gelatin-chitosan scaffold is not effective in repair of femoral bone defect.

Aim: Electromagnetic field as a physical stimulus, willingly or unwillingly are affected cellular processes. In this study morphology changing and proliferation rate of mesenchymal stem cells were investigated in presence of electromagnetic field (EMF) and nitric oxide (NO).Material and methods: The stromal stem cells were isolated from the Rat bone marrow and incubated. After several passages of the harvested cells, Deta-NO as a donor of nitric oxide was then added to cell culture. These cells were also treated by EMF (50 Hz and 20 mT). The MTT test was used for estimating proliferation rate of the cells. To estimate the proportion of the cell line in different phases of cell cycle due to treatment with NO and EMF, cellular DNA contents were measured by flow cytometry.Result: The results demonstrated decreasing proliferation rate of the stem cells exposed with EMF and NO compared with the control group. We found that the rate of decrease is highly related to the concentration of NO. The double treatment of EMF and nitric oxide was yielded to an obvious arrest in the cell cycle at G2/M phase. Nitric oxide associated with EMF also changed the cell morphology and increased the cell motilities.Conclusion: The changing of cell morphology and reduction of proliferation rate of the stem cell can be considered as a symptom showing the beginning of cell differentiation.

Aim: The aim of this study was to investigate the preventive role of vitaminE (Vit. E) on spermatogenesis indexes in rats following exposure to Bisphenol A.Materialand and Methods: Adult male wistar rats with the mean body weight of 231±10g were randomly divided into 4 groups (n=6): control, BPA (250mg/kg/day), Vit. E (150mg/kg/day) and BPA+Vit. E. Oral treatment was performed three times a week till 56 days. At the end of the treatment, the rats were killed and their left testis were weighed, fixed and stained using Heidenhain's Azan methed. Histological and morphometrical analysis of spermatogenesis was carried out. Data were analyzed using One Way ANOVA and the means were considered significantly different at p<0.05.Results: Testis weight (P<0.01) significantly decreased in the BPA group compared to the control. A significant decrease (P<0.001) in the mean diameter of seminiferous tubules and the germinal epithelium thickness and the indexes of tubular differentiation, spermiogenesis and meiosis were also observed in the testicular tissue of rats treated with BPA compared to the control ones. In the BPA+Vit. E group, the mentioned parameters increased significantly to the control level.Conclusion: Vitamin E can compensate for the undesired effects of BPA on spermatogenesis and therefore could be considered as a therapeutic supplement in the case of BPA toxicity.

Aim: Infertility is the failure of a couple to engender after endeavoring at least one year of unprotected intercourse. male factor infertility accounts for approximately 50 percent of causes. Tumor necrosis factor-a(TNF-a) is a multifunctional cytokine. TNF-a plays important role in the regulation of cellular processes related to spermatogenesis. There are two variants of the cell receptors that interacts with TNF-a. In the present study, the association of TNFR1 36A/G polymorphisms with idiopathic male infertility in the Guilan population was studied.Materials and Methods: This study consists of 106 infertile men and 114 fertile men as control group. Blood samples were taken and genomic DNA was extracted. Then genotypes and allele frequencies were assessed by PCR-RFLP method and the statistical analysis was performed by Med Calc software.Results: The frequencies of AA, AG, GG genotypes in patients were 41.5%, 7.5% and 50.9%, respectively and in controls were 45.6%, 33.3%, and 21.1%, respectively. The frequencies of A and G in patients were 0.45 and 0.55, respectively and in controls were 0.62 and 0.38, respectively. The results showed that there is a significant association between genotype frequency (p=0.002) and allele frequency (p=0.0004) in infertile and control groups.Conclusion: In conclusion, the subjects with G allele and GG genotype appears to be at greater risk of developing idiopathic infertility in Guilan province. Further studies with larger numbers of patients are required to elucidate the potential role of TNFR1 polymorphism in male infertility.