19 research outputs found
Changes in Plasma Level of oxidative Stress, hs-CRP and Anti-Hsp27 Titers in Stroke patients
No full textOverexpression of microRNA-16 declines cellular growth, proliferation and induces apoptosis in human breast cancer cells
No full textMicroRNAs (miRNA) are a large family of small single-stranded RNA molecules found in all multicellular organisms. Early studies have been shown that miRNA are involved in cancer development and progression, and this role can be done by working as an oncogenes and tumor suppressor genes, so manipulation of this molecules can be a promising approach in cancer therapy, and experimental results represented that the modification in breast cancer phenotype is possible by miRNA expression alteration. miR-16, which is located in 13q14 chromosome, plays critical roles as a tumor suppressor by targeting several oncogenes which regulate cell cycle and apoptosis. Hence, in the present study, we investigated whether miR-16 could decline growth and survival of MCF-7 cell line as model of human breast cancer. MCF-7 cell line was infected with lentiviruses containing miR-16 precursor sequence. The effects of ectopic expression of miR-16 on breast cancer phenotype were examined by cell cycle analysis and apoptosis assays. miR-16 cytotoxicity effect was measured by the MTT assay. We showed that the miR-16 overexpression reduces Cyclin D1 and BCL2 at messenger RNA (mRNA) and protein levels in MCF-7 cell line. In addition, this is found that enforced expression of miR-16 decreases cell growth and proliferation and induces apoptosis in MCF-7 cells. In conclusion, our results revealed that upregulation of miR-16 would be a potential approach for breast cancer therapy. © 2015, The Society for In Vitro Biology
Evaluation of the expansion of umbilical cord blood derived from CD133+ cells on biocompatible microwells
No full textBackground: Hematopoietic stem cell transplantation (HSCT) is a therapeutic approach for treatment of hematological malignancies and incompatibility of Bone marrow. Umbilical cord blood (UCB) has known as an alternative for hematopoietic stem/progenitor cells (HPSC) in allogeneic transplantation. The low volume of collected samples is the main hindrance in application of HPSC derived from umbilical cord blood. So, ex vivo expansion of HPSCs is the useful approach to overcome this restriction. The goal of using this system is to produce appropriate amount of hematopoietic stem cells, which have the ability of transplantation and long term haematopoiesis.
Material & Methods: In current study CD133+ cells were isolated from cord blood (CB). Isolated cells were seeded on microwells. Then expanded cells proliferation rate and ability in colony formation were assessed and finally were compared with 2 Dimensional (2D) culture systems.
Results: Our findings demonstrated that CD133+ cells derived from UCB which were cultivated on microwells had significantly higher rate of proliferation in compared with routine cell culture systems.
Conclusion: In Current study, it was shown that CD133+ cells’ proliferations which were seeded on PDMS microwells coated with collagen significantly increased. We hope that 3 dimensional (3D) microenvironment which mimics the 3D structure of bone marrow can solve the problem of using UCB as an alternative source of bone marrow
Calprotectin Pegylation Enhanced Its Physical and Structural Properties
No full textAbstract Calprotectin is member of the S-100 protein
family with a wide plethora of intra-and extracellular
functions. Anticancer activities, antimicrobial effects and
being a qualified disease marker are among the compelling
features of this protein to be used as a pharmaceutical
agent. However, there are several impediments to applications
of protein pharmaceuticals including: proteolytic
degradation, short circulating half-life, low solubility and
immunogenicity. Pegylation is a common bioconjugation
polymer capable of overcoming these drawbacks. Recombinant
expression and purification of calprotectin along
with its pegylation would result in enhanced pharmacodynamic
and pharmacokinetic properties. Our florescence
spectroscopy and far Ultraviolet-optical density results
indicate that pegylation altered the physical and structural
properties of the calprotectin to become in a more
stable and functionally active state. Due to enhanced
pharmacodynamic and pharmacokinetic properties of the
calprotectin via pegylation, this study would pave the way
for better in vitro and in vivo validations of calprotectin
applications in medical practice
Three-dimensional nanofiberous PLLA/PCL scaffold improved biochemical and molecular markers hiPS cell-derived insulin-producing islet-like cells
No full textNanofibrous scaffolds are considered as a new strategy for Type 1 diabetes mellitus therapy. We used a hybrid of poly-l-lactic acid (PLLA) and polycaprolactone (PCL) as three-dimensional (3D) culture models for differentiation of human induced pluripotent stem cells (hiPSCs) to beta islet-like cluster cell compared with routine culture (2D). Morphological changes of cells were checked by microscope. mRNA endodermal SOX-17 on day 7 and pancreatic gene markers Pdx1, glucagon and Glut2 were evaluated on day 23 by qPCR. As well as, insulin and C-peptide protein expression was evaluated by immunocytochemistry staining. In addition, insulin and C-peptide secretion in various glucose concentrations was evaluated by ELISA. Light and scanning electron microscopy (SEM) microscope showed changes in induced cells. In tandem, these modifications were more evident in 3 D culture. Pdx1, Glucagon and Glut2 markers in PLLA/PCL were significantly higher in 3 D culture. In addition, qualitative immunochemistry showed that insulin and C-peptide were expressed in 2 D and 3 D culture medium. Furthermore, evaluation of insulin and C-peptide clarified that secretion of these proteins in PLLA/PCL scaffold were statistically different in 2 D and 3 D strategies. These findings suggest that functional matured induction cells on PLLA/PCL scaffold can be used for islet beta cell therapy and regenerative medicine
Investigation of Leukemia Frequency in Children of Qazvin Province and its Correlation with Gender, Age, and Blood Groups between 2006-2016
No full textBackground: About 8 percent of all cancers in human population are related to leukemia and it is one of the most common malignancies in children. The aim of this study was to compare the prevalence of age, gender and blood group types with the frequency of leukemia among the children with leukemia in Qazvin province during the 2006 to 2016.Materials and Methods: This was a cross-sectional analysis. Investigated population was 110 children and adolescents under 18 years in the hospitals of Qazvin province. The date collecting method was through review of medical records of the patients and their analysis performed by using SPSS version 16.Results: According to data from this study, leukemia ALL-L1 is more frequent in Qazvin than other types of leukemia, and children with ages 0-5 years was more than other age groups. This disorder is more common in boys than girls, and among the patients, the people who has A and O blood groups, and Rh + are the most abundant.Conclusion: such factors like age, gender and blood groups can use as prognostic factors in children leukemia. So that leukemia in children less than 5 years old is more than any other age. In addition to that; the incidence of leukemia ALL-L1 reduced with increasing age in the general population in Qazvin and number of boys with leukemia is more than girls.</p
Hypoxia-induced miR-210 overexpression promotes the differentiation of human-induced pluripotent stem cells to hepatocyte-like cells on random nanofiber poly-L-lactic acid/poly (? -caprolactone) scaffolds
No full textAn alternative treatment to liver transplantation includes the use of differentiated stem cells. Hypoxia has been shown to endow human-induced pluripotent stem cells (hiPSCs) with enhanced hepatic differentiation. We have investigated a new strategy for hepatocyte differentiation from hiPSCs using a three-step differentiation protocol with lentiviral overexpression of hypoxia-microRNA-210 of cells grown on a hybrid scaffold. We analyzed the transduction of the miR-210 lentiviral and definitive endoderm and pluripotency gene markers, including SRY-box 17 (SOX17), forkhead box A2 (FOXA2), and octamer-binding transcription factor 4 (OCT-4) by Real-Time PCR and fluorescent microscope. The scanning electron microscopy (SEM) examined the 3D cell morphological changes. Immunocytochemistry staining was used together with assays for aspartate aminotransferase, alanine aminotransferase, and urea secretion to analyze hepatocyte biomarkers and functional markers consisting of a-fetoprotein (AFP), low-density lipoprotein (LDL) uptake, fat accumulation, and glycogen. The flow cytometry analyzed the generation of reactive oxygen species (ROS). Compared to cells transfected with the blank lentiviral vectors as a control, overexpressing miR-210 was at higher levels in hiPSCs. The expression of endodermal genes and glycogen synthesis significantly increased in the differentiated lentiviral miR-210 cells without any differences regarding lipid storage level. Additionally, cells containing miR-210 showed a greater expression of ALB, LDL, AST, ALT, urea, and insignificant lower AFP and ROS levels after 18 days. However, SEM showed no significant differences between cells under the differentiation process and controls. In conclusion, the differentiation of hiPSCs to hepatocyte-like cells under hypoxia miR-210 may be a suitable method for cell therapy and regenerative medicine
Laboratory Biochemical and Hematological Parameters: Early Predictive Biomarkers for Diagnosing Hepatitis C Virus Infection
No full textCorrelation between Methylation and Expression Level of P15 and P16 Genes during Differentiation of Cord Blood Stem Cells into Erythroid Lineage Mediated by Erythropoietin
No full textBackground: Several influential factors such as transcription factors and intracellular signaling components are involved in differentiation of stem cells into a specific lineage. P15 and p16 proteins are among these factors. Accumulating evidences has introduced the epigenetic as a master regulator of these factors during lineage specification. The main objective of this study is to determine the correlation between the expression level and methylation pattern of P15 and P16 genes in erythroid lineage after in vitro differentiation by erythropoietin (EPO).
Materials and Methods: The purified and expanded CD34+ cord blood stem cells were differentiated into erythroid lineage in the presence of EPO. DNA was isolated from both cord blood stem cells and differentiated cells. The Real-Time PCR performed using cDNA and the isolated DNA was used in methylation Specific PCR (MSP) reaction for methylation pattern analysis in both pre and post differentiation stages.
Results: The study demonstrated that P15 and P16 genes have partial methylation after erythroid differentiation by EPO. The Expression of P15 gene was higher after differentiation and the expression of P16 gene had a slightly decreased level in post differentiation stage.
Conclusion: Significant increase in P15 gene expression after differentiation to erythroid lineage, suggests the remarkable efficacy of this gene in erythroid function. According to upregulation of P15 gene after differentiation despite unchanged methylation status and slight down regulation of P16 gene with slight hyper-methylation of the gene it can be suggested that although the methylation can affects the expression level of P16 gene, the P15 gene is not affected by this mechanism during erythroid differentiation mediated by EPO
