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一种简单经济提取小鼠大量肾小球的实验方法
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作者 李艳秋 刘晓丹 +4 位作者 范秋灵 田淑艳 刘雪 王力宁 姚丽 《中国现代医学杂志》 CAS 2020年第7期10-13,共4页
目的为减轻实验者用磁珠提取肾小球的经济负担,对传统的心脏灌注肾脏提取方法进行改良。方法结扎远端腹主动脉及下腔静脉,结扎肠系膜上动脉和腹腔动脉,以减少磁珠分流。将磁珠工作液经静脉留置针灌注肾脏。加入相应体积胶原酶A,通过100... 目的为减轻实验者用磁珠提取肾小球的经济负担,对传统的心脏灌注肾脏提取方法进行改良。方法结扎远端腹主动脉及下腔静脉,结扎肠系膜上动脉和腹腔动脉,以减少磁珠分流。将磁珠工作液经静脉留置针灌注肾脏。加入相应体积胶原酶A,通过100μm筛网2次,加入蛋白裂解液,离心裂解吸出含蛋白上清进行肾小球蛋白的纯化、定量。结果与传统的心脏灌注肾脏提取方法比较,改良胸主动脉灌注肾脏提取肾小球方法磁珠用量仅为原来的1/35~1/40。在获取相同的蛋白情况下,提取肾小球的最佳浓度为4×10~6磁珠/ml,磁珠用量为20ml。通过该方法灌注分离的肾小球完整,纯度为(95.7±2.3)%。结论改良的胸主动脉灌注肾脏是一种简单经济实用磁珠提取肾小球的实验方法,可以快速获取大量完整的肾小球,蛋白质甚至RNA,为肾小球本身的研究提供新的技术支持。 展开更多
关键词 改良 肾小球 C57BL/6小鼠 NZB/WF1小鼠 磁珠
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大鼠骨髓单核细胞的快速分离方法及向破骨细胞的诱导分化 被引量:6
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作者 王礼宁 马勇 +3 位作者 郑苏阳 郭杨 袁翰 孙杰 《中国现代医学杂志》 CAS 北大核心 2017年第18期1-6,共6页
目的建立一种简便高效的体外分离大鼠骨髓单核细胞的方法,观察其体外生长特性,并诱导其分化为破骨细胞。方法无菌条件下分离出SD大鼠的胫骨和股骨,使用环氧树脂管(EP管)和移液枪头快速分离骨髓组织,再用红细胞裂解液去除红细胞。细胞培... 目的建立一种简便高效的体外分离大鼠骨髓单核细胞的方法,观察其体外生长特性,并诱导其分化为破骨细胞。方法无菌条件下分离出SD大鼠的胫骨和股骨,使用环氧树脂管(EP管)和移液枪头快速分离骨髓组织,再用红细胞裂解液去除红细胞。细胞培养过夜后收集悬浮细胞,加入巨噬细胞集落刺激因子(M-CSF)继续培养扩增后得到贴壁的骨髓单核细胞。观察骨髓单核细胞生长过程中的形态学特征;通过细胞计数试剂盒法(CCK-8)测绘细胞的生长曲线;用流式细胞仪检测细胞表面CD11b的表达;在加入M-CSF和核因子κB受体活化因子配基(RANKL)诱导分化后,用抗酒石酸酸性磷酸酶(TRAP)染色和降钙素受体(CTR)免疫荧光染色鉴定其是否能分化为成熟破骨细胞。结果通过该方法获得的大鼠骨髓单核细胞培养1 d后基本呈小圆形,杂细胞较少。3 d后细胞数目稍多,两端开始出现触角,5 d后数目增多,细胞呈椭圆形,两端触角明显;细胞的增殖依赖于M-CSF;流式结果显示,通过此方法获得的单核细胞纯度较高;TRAP染色和CTR免疫荧光染色结果提示,通过该方法获得的单核细胞可以诱导为成熟破骨细胞。结论通过EP管和移液枪头装置可快速分离获取单核细胞,获得的细胞表型稳定,适合用于骨代谢疾病的进一步研究。 展开更多
关键词 分离培养方法 破骨细胞 骨髓单核细胞 分化 骨代谢疾病
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Human podocyte injury in the early course of hypertensive renal injury 被引量:3
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作者 Da Sun Jiao-Jiao wang +5 位作者 Wei wang Juan wang li-ning wang Li Yao Ying-Hui Sun Zi-Long Li 《World Journal of Clinical Cases》 SCIE 2019年第22期3698-3710,共13页
BACKGROUND Hypertension is prevalent in the general population and is regarded as the second leading cause of renal damage and dysfunction,outnumbered only by diabetes.However,the mechanisms remain unclear.AIM To inve... BACKGROUND Hypertension is prevalent in the general population and is regarded as the second leading cause of renal damage and dysfunction,outnumbered only by diabetes.However,the mechanisms remain unclear.AIM To investigate podocyte injury induced by hypertension in the early course without massive proteinuria or renal dysfunction.METHODS The hypertension group comprised 18 patients with hypertension accompanied by microalbuminuria,diagnosed with hypertensive renal injury according to biopsy results.For a comparison of pathological changes in renal tissue,control group 1 comprised 10 healthy volunteers,and control group 2 comprised 16 patients who underwent surgery for renal trauma.RESULTS The hypertension group had significantly higher blood pressure(P=0.000)and microalbuminuria(P=0.000)compared with control group 1.In the hypertension group,urinary podocytes were detected following positive staining of podocytespecific nephrin and/or CD2-associated protein(CD2AP)in urine sediment.Podocyte foot process fusion and a significant decrease in nephrin and/or CD2AP expression in glomeruli were observed in the hypertension group compared with control group 2.This indicated that hypertension caused podocyte injury and detachment from the glomerular basement membrane,which was consistent with urinary detection of podocytes.CONCLUSION Our results suggest that podocyturia appears early in the course of hypertensive renal injury,and may be a sensitive marker for early prediction of hypertensive renal injury. 展开更多
关键词 PODOCYTE HYPERTENSION HYPERTENSIVE RENAL injury MICROALBUMINURIA NEPHRIN CD2-associated protein
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Spillover of international crude oil prices on China's refined oil wholesale prices and price forecasting:Daily-frequency data of private enterprises and local refineries
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作者 Xun-Zhang Pan Xi-Ran Ma +3 位作者 li-ning wang Ya-Chen Lu Jia-Quan Dai Xiang Li 《Petroleum Science》 SCIE CAS CSCD 2022年第3期1433-1442,共10页
Compared with retail prices of state-owned companies used in almost all existing studies,China’s refined oil wholesale prices of private enterprises and local refineries are more affected by the market and better ref... Compared with retail prices of state-owned companies used in almost all existing studies,China’s refined oil wholesale prices of private enterprises and local refineries are more affected by the market and better reflect the real supply-demand situation.For the first time,this paper applies own-monitored dailyfrequency wholesale prices of China’s private enterprises and local refineries during 2013-2020 to derive spillover effects of international crude oil prices on China’s refined oil prices through the VAR-BEKKGARCH(vector autoregression-Baba,Engle,Kraft,and Kroner-generalized autoregressive conditional heteroscedasticity)model,and then tries to forecast wholesale prices through the PCA-BP(principal component analysis-back propagation)neural network model.Results show that international crude oil prices have significant mean spillover and volatility spillover effects on China’s refined oil wholesale prices.Changes in crude oil prices are the Granger cause of changes in refined oil wholesale prices.With the improvement of China’s oil-pricing mechanism in 2016,the volatility spillover from the international crude oil market to China’s refined oil market gradually increases,and the BRENT price variation has an increasing impact on the refined oil wholesale price variation.The PCA-BP model could serve as a candidate tool for forecasting China’s refined oil wholesale prices. 展开更多
关键词 Own-monitored daily-frequency data Refined oil wholesale prices Spillover effects VAR-BEKK-GARCH PCA-BP neural Network
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Isolation and Identification of Bioactive Constituents from Stem Barks of Illicium difengpi 被引量:4
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作者 Yong-zhi He Eric Kibagendi Osoro +2 位作者 Sivoko Imbenzi Palmer li-ning wang Naji Said Aboud 《Chinese Herbal Medicines》 CAS 2014年第1期76-79,共4页
Objective To isolate and identify bioactive constituents from the stem barks of Illicium difengpi.Methods The chemical constituents were isolated and purified by repeated silica gel,Sephadex LH-20,recrystallization,an... Objective To isolate and identify bioactive constituents from the stem barks of Illicium difengpi.Methods The chemical constituents were isolated and purified by repeated silica gel,Sephadex LH-20,recrystallization,and preparative HPLC techniques.The structures of the compounds were identified on the basis of spectral data including NMR,MS,and IR.Results Two sesquiterpene lactones,majucin(1)and anisatin(2),two steroids,β-sitosterol(3)and daucosterol(4),three carboxylic acids,2-ethyldecanoic acid(5),shikimic acid(6),and 3,4-dihydrobenzoic acid(7),and a flavonoid,quercetin(8),were successively isolated from the stem barks of I.difengpi.Conclusion Apart from compound 3,other seven compounds are reported in this plant for the first time.The results suggested that the current studies on I.difengpi is still far from being well known and therefore more studies need to be done for better understanding of this plant. 展开更多
关键词 中药学 中草药 化学成分 药效
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