BACKGROUND:To build up an effective method of isolating and culturing granule cels is a pivotal step to enhance fertilization-embryo transfer rate. Current studies mainly focus on the isolation methods of human ovarian granulosa cels rather than cel counting, purity and subsequent growth. OBJECTIVE: To establish the effective methods of isolating, purifying and culturing human ovarian granulosa cels in vitro. METHODS: Folicular fluid was harvested from women undergoing fertilization-embryo transfer procedures. Human ovarian granulosa cels were obtained from the folicular fluid by lysis treatment, precipitation method or density gradient centrifugation. Granulosa cel mucus masses were digested with type I colagen enzyme or hyaluronidase and then cultured in the culture medium with or without autologous folicular fluid. RESULTS AND CONCLUSION: Lysis treatment yielded the largest amount of granulosa cels compared to the precipitation method and density gradient centrifugation (P > 0.05,P < 0.05, respectively). Cels prepared by the three methods showed the same cel viability. After 24 hours of culture, the precipitation method obtained the largest amount of adherent granulosa cels (P < 0.05); and the density gradient centrifugation obtained the least amount of cels (P < 0.05). Compared with type I colagen enzyme, hyaluronidase took less time to digest the cels thoroughly. Autologous folicular fluid could promote the growth and survival of granulosa cels. These findings indicate that the precipitation method, though time-consuming, can obtain the highest cel viability and harvested the largest amount of granulosa cels after culture; hyaluronidase is more suitable for digesting granulosa cel mucus mass than type I colagen enzyme; autologous folicular fluid added into the culture medium is more conducive to granulosa cel growth.

Objective: To preliminarily evaluate coronary heart disease (CAD) by dual-source CT vascular functional imaging in relevant patients. Methods: A total of 200 patients with suspected non-ST elevation acute coronary syndrome (NSTE-ACS) in our hospital from 2014-09 to 2015-10 were enrolled, 57 of them received dual-source CT angiography (DSCTA) and diagnosed for critical value of left anterior descending (LAD) stenosis; the patients were further examined by selective coronary angiography (SCA) within 1 week to conifrm the degree of stenosis. Meanwhile, fractional lfow reserve (FFR) was measured and taking FFR 0.8 as cut off point, the patients were divided into 2 groups: FFR<0.8 group,n=27 and FFR≥0.8 group,n=30. The values of left ventricular anterior wall, side wall, left ventricular cavity and the segmental thickness in diastolic and systolic stages were measured; relative CT value between ventricular anterior wall and side wall was compared, myocardium thickness at the end-diastolic stage was also compared. Results:①In FFR<0.8 group, compared with the side wall, anterior wall had decreased relative CT value (P=0.000), myocardium thickness at the end-diastolic stage (P=0.000) and myocardial wall thickening rate (P=0.001).②In FFR≥0.8 group, compared with the side wall, anterior wall had decreased relative CT value (P=0.000), myocardium thickness at the end-diastolic stage (P=0.018), while similar myocardial wall thickening rate (P=0.186).③Compared with FFR≥0.8 group, the patients in FFR<0.8group presented reduced relative CT value in anterior wall (P<0.05) and myocardial wall thickening rate (P<0.001), while similar myocardium thickness at the end-diastolic stage (P=0.964). Conclusion: CT information may provide the reference value for treating patients in clinical practice.