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Objective To investigate the effects of Rho-associated coiled-coil-forming kinase (ROCK) inhibitor Y27632 on the development of mouse preantral follicles in vitro. Methods The ovaries of 12.5-day-old mice were collected and single preantral follicle was isolated by mechanical method to culture Nunclon Sphera 96-well U-bottom 3D cell culture plate in vitro. Set up a control group and treated group containing Y27632, and evaluate the overall development of the follicles through follicular morphology, follicle diameter, the number of mature follicles, and changes in oocyte spindles. The expression of oocytes [bone morphogenetic protein 15(BMP 15), growth differentiation factor 9(GDF9)], granulosa cells [follicle stimulating hormone receptor (FSHR)] and apoptosis related genes (Bad, Bax and Caspase-3) were also assessed by the Real-time PCR technique. Meanwhile, the follicle viability was detected by follicular activity test during follicular development. Results Rock inhibitor Y27632 had no significant effect on the growth diameter and basic development indicators (follicle survival rate, cavity formation rate and maturation rate) (P>0.05). And abnormal spindle assembly occurred during follicle development in the control group. After 8 days of culture, compared with the control group, the granulosa cell-specific gene FSHR was up-regulated in the experimental group; As well as the oocyte-specific genes BMP 15 and GDF9 were up-regulated, while forkhead box 03(Fox03) was down-regulated; The apoptosis genes Bax, Bad and Caspase-3 were also showed down-regulated. And the granulosa cell apoptosis in the experimental group was significantly less than that in the control group. Conclusion ROCK inhibitor Y27632 can inhibit the apoptosis of granulosa cells and prevents the abnormal assembly of oocyte spindles to improve the quality of follicle development in vitro.
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Objective To investigate the number and activity changes of endothelial progenitor cells(EPCs) in hemodialysis patients,and explore its correlation with the risk factors of coronary heart disease.Methords Total mononuclear ceils(MNCs) were isolated from peripheral blood of patients with chronic renal failure in long-term hemodialysis and from a normal control group by Ficoll density gradient centrifugation and then were plated on humanfibronectin-coated dishes.Mter 7 days of culture,EPCs were characterized as adherent cells by double staining with FITC-UEA-I and DI-LDL,and were further identified by the expression of CD34,CD133 and KDR with flow cytometry.EPCs migration was determined with modified Boyden chamber assay.EPCs adhesive assay was performed by replanting EPCs on humanfibronectin-coated plates and then counting the adherent cells.The relationship of the EPCs' number and activity with Framingham Risk Score of ten years was also be assessed.Results Number of EPCs and the migratory & adhesive capacity were significantly lower in patients than in the control(P
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<p><b>OBJECTIVE</b>To find a better index for detecting myocardial lesion in patients with acute ammonia poisoning.</p><p><b>METHODS</b>Cardiac troponin T (cTnT), creatine kinase (CK), creatine kinase-MB isoenzyme (CK-MB), and electrocardiogram (ECG) were determined and compared in patients with acute ammonia poisoning and healthy controls.</p><p><b>RESULTS</b>In severe ammonia poisoning patients, the levels of cTnT [(1.285 +/- 2.650) micro g/L], CK [(257.636 +/- 362.719) IU/L], CK-MB [(20.909 +/- 19.770) IU/L] were significantly higher than those in healthy control [(0.035 +/- 0.014) micro g/L, (82.050 +/- 36.302) IU/L, (8.350 +/- 2.455) IU/L respectively, P < 0.05]. The levels of cTnT, CK and CK-MB in mild ammonia poisoning patients [(0.039 +/- 0.016) micro g/L, (78.200 +/- 28.401) IU/L and (8.467 +/- 2.669) IU/L respectively], and moderate ammonia poisoning patients [(0.054 +/- 0.043) micro g/L, (99.786 +/- 71.941) IU/L and (9.429 +/- 3.857) IU/L were not significantly different from those of healthy controls (P > 0.05)]. cTnT positive detection rate (68.2%) was significantly higher than CK (36.4%), CK-MB (31.2%) and ECG (31.2%) (P < 0.05).</p><p><b>CONCLUSION</b>cTnT is a better index to detect myocardial lesion in severe ammonia poisoning.</p>