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Objective:To investigate the protective effect and mechanism of miR-30c targeting Wnt/β-catenin signal on the proliferation and apoptosis of human retinal endothelial cells (HRECs) induced by high glucose.Methods:Human retinal endothelial cells (HRECs) were cultured and given normal concentration (control group) and high concentration glucose (high glucose group) respectively. According to the experimental design, miR-30c mimic, negative control (miR-NC), Wnt1 overexpression vector (pcDNA3.1-Wnt1) and empty vector (pcDNA3.1) were transfected respectively. RT-PCR was used to detect the expression level of miR-30c in each group. Western blot was used to detect the expression levels of Wnt1, β-catenin and GSK-3β protein in each group. The dual luciferase experiment verified the targeting relationship of miR-30c to Wnt1. Thiazole blue method was used to detect the proliferation activity of each group. Flow cytometry was employed to detect the level of apoptosis of each group of cells.Results:Compared with the control group, the expression of miR-30c in the high glucose group was significantly reduced [ (0.94±0.11) vs (0.32±0.06), P<0.001]; compared with the control group, the cell proliferation activity of the high glucose group was significantly reduced, and the apoptosis rate was significantly increased [ (0.75±0.08) vs (0.13±0.04), (3.53±0.29) % vs (14.89±0.94) %, P<0.001]; compared with the high glucose+miR-NC group, the cell proliferation activity of the high glucose+miR-30c group was significantly increased, and the apoptosis rate was reduced [ (0.14±0.04) vs (0.64±0.06), (14.14±0.86) % vs (6.28±0.45) %, P<0.001]; compared with the miR-NC group, the luciferase activity of the miR-30c group co-transfected with WT-Wnt1 was significantly reduced [ (0.97±0.09) vs (0.26±0.03), P<0.001]; compared with the control group, the protein expression of Wnt1, β-catenin and GSK-3β in the high glucose group were significantly increased [ (0.43±0.05) vs (1.02±0.09), (0.25±0.04) vs (0.82±0.10), (0.39±0.04) vs (0.76±0.11), P<0.001]; compared with the high glucose+miR-NC group, the protein expression of Wnt1, β-catenin and GSK-3β in the high glucose+miR-30c group were significantly reduced [ (1.04±0.10) vs (0.68±0.06), (0.79±0.09) vs (0.34±0.05), (0.74±0.12) vs (0.48±0.06), P<0.001]; compared with the high glucose+miR-30c group, the cell proliferation activity was significantly reduced in the high glucose+miR-30c+pcDNA3.1-Wnt1 group, and the apoptosis rate was significantly increased [ (0.66±0.07) vs (0.31±0.05), (4.26±0.57) % vs (9.75±0.85) %, P<0.001]. Conclusion:miR-30c may negatively regulate the Wnt/β-catenin signaling pathway, promote cell proliferation, and inhibit cell apoptosis induced by high glucose.
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Objective To investigate the impact of bone marrow mesenchymal stem cell transplantation on a rat model of experimental autoimmune uveitis (EAU) and analyze its immune regulatory mechanisms in vivo. Methods Eighteen Lewis rats were randomly divided into three groups: model control group, intervention group and normal control group, six animals in each group. Human retinal S-antigen peptide (HS-AgP35, 1 mg/ml) was mixed and emulsified with complete Freund's adjuvant and injected into hind foot pad of rats on the first and eighth day to establish the animal model of EAU. For bone marrow mesenchymal stem cell transplantation, 1 ml of cell suspension (2 × 106 cells/ml) was injected into tail vein of the intervention group rats on the first day when the emulsified S-antigen was injected. EAU manifestation, pathological change and IFN-γ level were evaluated and compared among those three groups after two weeks. Results No abnormal signs were found in the eyes of rats in normal control group. The manifestation grading of the intervention group (two rats at grade 0, three rats at grade 0.5, one rat at grade one) was significantly different from the model control group (one rat at grade one, one rat at grade two, three rats at grade three, one rat at grade four) (P=0. 015). The retina of rats in normal control group was ordinary under light microscope. The histopathological grading of the intervention group (one rat at grade 0, four rats at grade 0.5, one rat at grade one) and the model control group (four rats at grade three, two rats at grade four) was also statistically different (P＜0. 01). Furthermore, the IFN-γ level in peripheral blood of the intervention group rats declined significantly compared to the model control group (t=9. 057 4, P=0. 01). Conclusions Bone marrow mesenchymal stem cells can inhibit EAU significantly,possibly by lowering the level of IFN-γ, thereby reduce the severity of uveitis and improve the condition of uveitis in rats.
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<p><b>OBJECTIVE</b>To investigate the effects of baicalin against Candida albicans germ tube formation, and adherence to buccal epitherial and vaginal epitherial cells.</p><p><b>METHOD</b>Various concentrations of baicalin (100, 50, 10 mg x L(-1)) were incubated with C. albicans suspension, the mixed suspension of C. albicans and human buccal epitherial cells, the mixed suspension of C. albicans and vaginal epitherial cells, respectively. The effects of baicalin on C. albicans germ tube formation, and adherence to buccal epitherial and vaginal epitherial cells were then assessed microscopically.</p><p><b>RESULT</b>All concentrations of baicalin could inhibit C. albicans germ tube formation, and adherent to buccal epitherial and vaginal epitherial cells,while there was no significant difference between standard and clinical strains.</p><p><b>CONCLUSION</b>Baicalin could inhibit C. albicans germ tube formation, and adherence to buccal epitherial and vaginal epitherial cells.</p>