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AIM:To establish an effective method for purification and culture of human cumulus cells(CCs) in vitro,and to compare the characteristics between CCs and mural granulosa cells(MGCs).METHODS:Follicular fluid and cumulus complex from the patients undergoing intracytoplasmic sperm injection were collected.CCs were mechanically cut from cumulus complex and then directly inoculated on a Petri dish, and MGCs were obtained from follicular fluid through density gradient centrifugation.The expression of follicle stimulating hormone receptor(FSHR)was determined by immunofluorescence.The cell growth curves were measured by CCK-8 assay.The secretion of estrogen was detected by ELISA.RESULTS:After incubated for 24 h, the adherence of CCs was observed.CCs and MGCs had similar growth characteristics and FSHR expression.The similar cell growth curves were observed by CCK-8 assay and the results of ELISA showed that they had comparable secretion of estrogen.CONCLUSION:Direct culture of CCs mechanically cutting from cumulus complex is an effective method.CCs had similar growth characteristics,growth curves and secretion of estro-gen to MGCs cultured in vitro and could be a substitutive source of granulosa cell subsets.
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AIM To evaluate saponins in Paris Polyphylla Smith var.chinensis (Franch.) Hara from twentyone growing areas.METHODS HPLC was adopted in the content determination of pennogenins { pennogenin-3-O-β-D-glu (1→3) [α-L-rha (1→2)]-β-D-glu (PGGR),polyphyllinsⅥ,Ⅶ,H} and dioscins (gracillin,polyphyllins Ⅰ,Ⅱ,Ⅴ),then SPSS 22.0 software was used for principal component analysis and cluster analysis.RESULTS Pennogenin was the main kind of Paris Polyphylla var.chinensis saponins,and PGGR,polyphyllins Ⅰ,Ⅴ,Ⅶ,H were the main effective constituents of this medicinal material.The comprehension scores of seven growing areas were more than zero,which was in sequence of Bazhong ＞ Qingchuan ＞ Yanjin ＞ Tongjiang ＞ Nanchuan ＞ Chongzhou ＞ Hongya.Twenty-one batches of samples were divided into four types,which was similar to cluster analysis results.CONCLUSION Bazhong,Qingchuan,Chongzhou,Hongya,Tongjiang in Sichuan,Yanjin in Yunnan,and Nanchuan in Chongqing are suitable for cultivating Paris Polyphylla var.chinensis,which can guarantee the stability of saponins.
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Objective To investigate the mechanism of reverse redistribution (RR) on dipyridamole 201Tl myocardial perfusion studies in the patients with coronary artery spasm. Methods Twenty-six patients with coronary artery spasm and presented as RR on dipyridamole 201Tl myocardial perfusion studies were enlisted as RR group, while other 16 patients with no coronary artery stenosis nor RR were enlisted as control group. Dipyridamole test was repeated during coronary angiography. Corrected thrombolysis in myocardial infarction (TIMI) frame count (CTFC) and TIMI myocardial perfusion grade (TMPG) were measured at RR related and non-RR related coronary arteries before and after dipyridamole infusion respectively.All of the data were analyzed by Student's t-test orχ2-test and correlation analysis. Results Coronary artery angiography showed slower blood flow and lower myocardial perfusion in RR related vessels when compared with non-RR related vessels in RR group, but there was no significant difference among the main coronary arteries in control group. The perfusion defects of RR area at rest were positively related to slowerblood velocity at corresponding coronary arteries ( r = 0.79, t = 10.18, P ＜ 0.001 ). In RR related vessels,CTFC were (36 ±6) frames and (26 ±7) frames (t =4.15, P ＜0.01 ), while TMPG were (2.02 ±0.39)grades and (2.92 ± 0.12) grades ( t = 2.25, P ＜ 0.05 ) before and after dipyridamole infusion, respectively.In non-RR related vessels, CTFC were (29 ±7) frames and (25 ±5) frames (t =2.31, P ＜0.05), while TMPG were (2.56 ± 0.31 ) grades and (2.96 ± 0.06) grades ( t = 2.17, P ＜ 0.05 ) before and after dipyridamole infusion, respectively. However, there were no significant changes of CTFC and TMPG before and after dipyridamole infusion in control group ( t = 0.932, 0.867, respectively, both P ＞ 0.05 ). Conclusion RR is related to the decreased blood flow and myocardial perfusion induced by coronary artery spasm at rest,which may be improved by stress test such as intravenous dipyridamole infusion.