目的构建小鼠胚胎成纤维细胞(MEF)复制性衰老模型,检测衰老溶酶体相关功能改变,为衰老相关溶酶体疾病提供有效的细胞模型。方法通过酶消化法分离提取MEF,体外连续传代培养构建MEF复制性衰老模型,实时定量PCR(qPCR)检测衰老标志物p16和p...目的构建小鼠胚胎成纤维细胞(MEF)复制性衰老模型,检测衰老溶酶体相关功能改变,为衰老相关溶酶体疾病提供有效的细胞模型。方法通过酶消化法分离提取MEF,体外连续传代培养构建MEF复制性衰老模型,实时定量PCR(qPCR)检测衰老标志物p16和p21,衰老相关β⁃半乳糖苷酶(SA⁃β⁃gal)染色方法进一步验证衰老状态;通过溶酶体探针类染料LysoTracker Red DND⁃99探针及LysoSensor Yellow/Blue DND⁃160双激发探针追踪酸性溶酶体定性检测溶酶体酸碱性及定量检测溶酶体pH值;通过偶联自淬BODIPY®染料的牛血清白蛋白(DQ⁃BSA)检测衰老溶酶体的降解能力。结果qPCR和SA⁃β⁃gal染色结果显示,MEF复制性衰老模型构建成功。与年轻P3代MEF相比,衰老P9代MEF溶酶体的酸性明显丧失,差异有统计学意义(P<0.0001),衰老P9代MEF溶酶体的降解能力明显降低(P<0.05)。结论成功构建MEF复制性衰老细胞模型,并证明衰老MEF溶酶体的酸性丧失以及降解能力下降。展开更多
We set forth the macroeconomics evaluation of floods （MEF） model, a new model to assess and evaluate the impact of floods on GNP growth. This model points to a new, more concrete approach to measure the economic imp...We set forth the macroeconomics evaluation of floods （MEF） model, a new model to assess and evaluate the impact of floods on GNP growth. This model points to a new, more concrete approach to measure the economic impact of floods. Up to now, the measurement has been subject to a great deal of uncertainty. The main contribution of the model is to significantly reduce this uncertainty by mea- suring the impact with four well-defined, economically intuitive indicators. To illuminate and demonstrate its promise, we employ the model to evaluate and analyze the impact of two major floods on the economy of the People＇s Republic of China. Our MEF-model analysis indicates that the floods of Zhangshu and Jiangxi in 2010 caused greater economic damage than the floods of Central South China in 1931. Going forward, MEF-model simulations are a valuable tool for estimating the effect of potential future floods in the PRC and elsewhere.展开更多
文摘目的构建小鼠胚胎成纤维细胞(MEF)复制性衰老模型,检测衰老溶酶体相关功能改变,为衰老相关溶酶体疾病提供有效的细胞模型。方法通过酶消化法分离提取MEF,体外连续传代培养构建MEF复制性衰老模型,实时定量PCR(qPCR)检测衰老标志物p16和p21,衰老相关β⁃半乳糖苷酶(SA⁃β⁃gal)染色方法进一步验证衰老状态;通过溶酶体探针类染料LysoTracker Red DND⁃99探针及LysoSensor Yellow/Blue DND⁃160双激发探针追踪酸性溶酶体定性检测溶酶体酸碱性及定量检测溶酶体pH值;通过偶联自淬BODIPY®染料的牛血清白蛋白(DQ⁃BSA)检测衰老溶酶体的降解能力。结果qPCR和SA⁃β⁃gal染色结果显示,MEF复制性衰老模型构建成功。与年轻P3代MEF相比,衰老P9代MEF溶酶体的酸性明显丧失,差异有统计学意义(P<0.0001),衰老P9代MEF溶酶体的降解能力明显降低(P<0.05)。结论成功构建MEF复制性衰老细胞模型,并证明衰老MEF溶酶体的酸性丧失以及降解能力下降。
文摘We set forth the macroeconomics evaluation of floods （MEF） model, a new model to assess and evaluate the impact of floods on GNP growth. This model points to a new, more concrete approach to measure the economic impact of floods. Up to now, the measurement has been subject to a great deal of uncertainty. The main contribution of the model is to significantly reduce this uncertainty by mea- suring the impact with four well-defined, economically intuitive indicators. To illuminate and demonstrate its promise, we employ the model to evaluate and analyze the impact of two major floods on the economy of the People＇s Republic of China. Our MEF-model analysis indicates that the floods of Zhangshu and Jiangxi in 2010 caused greater economic damage than the floods of Central South China in 1931. Going forward, MEF-model simulations are a valuable tool for estimating the effect of potential future floods in the PRC and elsewhere.
