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The human resistin expression vector was constructed and transfected into cells to observe its effects on glucose uptake in adipocytes,cell proliferation and differentiation.The results suggested that human resistin impaired glucose uptake in adipoeytes via stimulating proliferation of preadipocytes and suppressing adipocyte differentiation.Metformin reversed the inhibition imposed by resistin on adipogenesis.
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Objective To investigate the inhibitory effect of silencing survivin gene with siRNA on the growth of gastric cancer cells and its mechanism,and provide evidence in treatment for gastric cancer.Methods DNA template coding survivinspecific siRNA was designed and synthesized.Two recombinant plasmids (pGCsilencerU6/GFP/survivin-siRNA-1 and-2) were constructed.The gastric carcinoma cel1 line SGC-7901 were divided into three groups: liposome-treated control group,empty plasmid-transfected control group and survivin-siRNA-1 transfected group.In order to observe the effect of survivin-siRNA,the expressions of survivin mRNA and protein were detected by RT-PCR and Western blotting,respectively.Methyl thiazolyl tetrazolium(MTT) assay was applied to determine the cell growth status.Apoptotic rates were evaluated by flow cytometry(FCM).Results The results of Western blotting and RT-PCR indicated that the inhibitory rates of protein and mRNA in pGCsilenerU6/GFP/survivin-siRNA-1 transfected group(78.25% and 88.75%) were higher than those in liposome-treated control group(5% and 2%) and empty plasmid-transfected control group(1% and 6%)(P
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Objective:The experiment researched relation between applying p38MAPK inhibitor(SB203580) and pulmonary injury during CPB.Methods:18 pigs randomly distributed three groups(n=6). We perfused the lung of the second group with SB203580 during CPB. The first group(n=6) and the second group(n=6) are control group. Lung samples were collected. Activated forms of P38 were measured by Western blot. Pulmonary injury was determined by histology. Tissue water percentage was measured. Pulmonary function was measured.Results:Phospho-P38 of the first group were increased compared with both the second group and the third group(P
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Objective:To study the relationship between human leukocyte antigen(HLA) typing, HLA crossreaction antigen group(CREGs) mismatching and early rejection after cadaver donor kidney transplantation.Methods:Donor and recipient HLA class Ⅰ typing was performed with Paul Terasaki monoclonal antibody tray and HLA class Ⅱ gene typing with Micro-sequence-specific-primers(Micro-SSP). PRA was detected by using Paul Terasaki mixture antigen tray.Results:The cases of 6, 5, 4, 3, 2, 1, 0 HLA mismatching(MM) in 131 recipients with negative PRA were respectively 0, 4, 26, 49, 33, 15, 4. The incidence of early rejection were 0,25%,23.1%,14.3%,12.1%,6.7%,0. No recipient with 5 and 6 CREGs MM. The case of 4,3,2,1,0 CREGs MM, the early rejection were respectively 28.6%,22.9%,9.5%,6.9%,5.5%.Conclusion:HLA typing and HLA crossreactive antigen group(CREGs) matching play an important role in selecting suitable donor, reducing early rejection, improving the effect of kidney transplantation.
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Objective :To study mutation of Ps3 gene and its relationship with histologic grade and Dukes' stage in colorectal cancer. Methods :PCR-SSCP was used to detect mutations of Ps3 in 30 colorectal cancer patients. Results: It was found that 6 mutations (20. 0%) in the coding region of P53 gene. One mutation was located in exon4; 5 mutations were located in exon 5; none of mutation was found in exon 2 and 3. According to the histologic grade of of colorectal cancer,the frequencies of P53 gene mutation were 1/8 (12. 5%) in the well differentiated and in the moderately differentiated as well,3/10 (30. 0%) in the poorly differenctiated and 1/4 (25.0%) in the undifferentiated. In clinic, the frequencies of P53 gene mutation were associated with Dukes stage: 1/10 (10. 0 % ) in B stage, 2/12 (16. 7%) in C Stage,3/8(37. 5%) in D stage. Conclusion:The mutation of P53 gene is related to Dukes＇ stage,and the P53 gene mutation hardly occurs in exon 2,3 in coloretal cancer.
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AIM: To study apoptotic injury induced by aeactive oxygen species - hydrogen peroxide (H2O2) on cardiac myocytes. METHODS: Cultured rat neonatal cardiac myocytes were treated with H2O2 of various concentra- tion to observe apoptoitic injury of cardiomyocytes by agarose gel electrophoresis, Giemsa - stained smears of cell, and flow cytometry, meanwhile lactate dehydrogenas (LDH) and malondialdehyde (MDA) were determined to assess the effect of H2O2 on lipid peroxidation and permeability of the plasma membrane. RESULTS: 5 mmol/L H2O2 caused culted cardiomyocytes apoptotic morophological characteristics, including nucleosomal DNA fragmentation in my- ocytes by agarose gell electrophoresis (DNA ladder), cell shrinkage, nuclear condensation, and chromatin margin by Giemsa-stained cell smears, and aneuploid peak (AP) - apoptotic bodies occurence by flow cytometry. CONCLU- SONS: H2O2 - induced apotosis in myocytes was a the - and concentration - dependent process, Treatment with low concentration of H2O2(10 mmol/L) rapidly induced a necrotic form of death characterized by smeared patterns of DNA digestion on agarose gel electrophoresis and lethal membrane disuption (as measured by LDH release). Exposure of 5 - 10 mmol/ L H2O2 induced cardiomyocytes apoptosis concurrently with biochemical changes of LDH and MDA increase in the medium.
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Objective:The human heparin-binding growth factor Midkine(hMK) gene was constructed and expressed in E.coli, and the activity of MK protein was tested.Methods:The MK gene was amplificated from the tissue of human embryo kidney by RT-PCR techniques. The MK fragment without signal peptide was inserted into plasmid pET30a. The gene was expressed in E.coli, the protein was purificated by affinity chromatography, and the activity was tested by ~3H-TdR method.Results:The sequence of hMK in our experiment was as same as the sequence published in Genebank. The activity assay was shown high activity of MK protein in our experiment.Conclusion:The expression carrier of hMK was established successful and it can expression high activity hMK protein in E.coli.