The mechanism of peroxynitrite (ONOO -)induced [Ca 2+ ] i increase in single MN9D cell (Dopaminergic neuroblastoma cell line) was studied by using Fura2 microfluorometric technique. The results show that ONOO - caused...The mechanism of peroxynitrite (ONOO -)induced [Ca 2+ ] i increase in single MN9D cell (Dopaminergic neuroblastoma cell line) was studied by using Fura2 microfluorometric technique. The results show that ONOO - caused a rapid increase of [Ca 2+ ] i when ONOO - was puffed to the cell. Removing Ca 2+ from the bath or using calcium channel antagonist (CdCl 2, Nifedipine) greatly inhibited the [Ca 2+ ] i increase induced by ONOO -, suggesting that the opening of LCa 2+ channel makes a great contribution to the [Ca 2+ ]\-i increase. The effect of sulfhydryl reductive agent (DTT) on ONOO -induced [Ca 2+ ]\-i increase suggests that ONOO -activating LCa 2+ channel is partly related to its oxidative speciality.展开更多
The microstructure, composition and shape of precipitated phase under as-cast and finished product state of 5Cr21Mn9Ni4N steel with different rare earth （RE） amount were studied. Mechanical properties of 5Cr21Mn9Ni4...The microstructure, composition and shape of precipitated phase under as-cast and finished product state of 5Cr21Mn9Ni4N steel with different rare earth （RE） amount were studied. Mechanical properties of 5Cr21Mn9Ni4N steels withont RE addition and with RE added by 0. 2% in mass percent were tested respectively. The results indicate that the solid solution amount of RE is about 10^-6 -10^-5 order of magnitude in 5Cr21Mn9Ni4N steel. Dendrite of as-cast condition is refined obviously and dimension of interstitial phase is shortened when RE is added by 0.10%-0.20%. But the microstructure will be coarser if surplus RE is added. Precipitated phase under finished product state distributes evenly in nearly same size with RE added by 0. 2% which leads to a largely improved high temperature mechanical property.展开更多
目的探讨α-突触核蛋白（α-synuclein）各结构域与MN9D细胞线粒体的关系。方法用PCR方法获得α-synuclein/1~65（N），α-synuclein/61~95（A）及α-synuclein/96~140（C）基因片段，克隆入真核表达质粒pLNCX2，经测序正确后以脂质体转...目的探讨α-突触核蛋白（α-synuclein）各结构域与MN9D细胞线粒体的关系。方法用PCR方法获得α-synuclein/1~65（N），α-synuclein/61~95（A）及α-synuclein/96~140（C）基因片段，克隆入真核表达质粒pLNCX2，经测序正确后以脂质体转染PT-67细胞，挑取单克隆并扩增，收集上清感染MN9D细胞，分别用Real Time PCR检测细胞基因表达水平，免疫组织化学染色检测蛋白表达，激光扫描共焦显微镜检测蛋白与线粒体共定位，流式细胞术检测细胞状态及线粒体膜电位状态。结果成功构建了pLNCX2/N、pLNCX2/NAC及pLNCX2/C基因片段的重组真核表达质粒，获得了可稳定表达α-synuclein各基因片段的MN9D细胞株。通过激光扫描共焦显微镜观察，可见α-synuclein/N端与线粒体存在共定位关系；α-synuclein/NAC主要在核内聚集表达；α-synuclein/C端在胞质和胞核内均有表达。JC1染色流式细胞术检测显示，过表达α-synuclein/N实验组细胞线粒体膜电位降低。结论α-synuclein的N端可能定位于线粒体，并参与调节线粒体功能；α-synuclein/NAC虽具有疏水性但却能穿过核膜，聚集在核仁周围；α-synuclein的C端定位于胞质和核内可能参与多种细胞功能。展开更多
文摘The mechanism of peroxynitrite (ONOO -)induced [Ca 2+ ] i increase in single MN9D cell (Dopaminergic neuroblastoma cell line) was studied by using Fura2 microfluorometric technique. The results show that ONOO - caused a rapid increase of [Ca 2+ ] i when ONOO - was puffed to the cell. Removing Ca 2+ from the bath or using calcium channel antagonist (CdCl 2, Nifedipine) greatly inhibited the [Ca 2+ ] i increase induced by ONOO -, suggesting that the opening of LCa 2+ channel makes a great contribution to the [Ca 2+ ]\-i increase. The effect of sulfhydryl reductive agent (DTT) on ONOO -induced [Ca 2+ ]\-i increase suggests that ONOO -activating LCa 2+ channel is partly related to its oxidative speciality.
文摘The microstructure, composition and shape of precipitated phase under as-cast and finished product state of 5Cr21Mn9Ni4N steel with different rare earth （RE） amount were studied. Mechanical properties of 5Cr21Mn9Ni4N steels withont RE addition and with RE added by 0. 2% in mass percent were tested respectively. The results indicate that the solid solution amount of RE is about 10^-6 -10^-5 order of magnitude in 5Cr21Mn9Ni4N steel. Dendrite of as-cast condition is refined obviously and dimension of interstitial phase is shortened when RE is added by 0.10%-0.20%. But the microstructure will be coarser if surplus RE is added. Precipitated phase under finished product state distributes evenly in nearly same size with RE added by 0. 2% which leads to a largely improved high temperature mechanical property.
文摘目的探讨α-突触核蛋白（α-synuclein）各结构域与MN9D细胞线粒体的关系。方法用PCR方法获得α-synuclein/1~65（N），α-synuclein/61~95（A）及α-synuclein/96~140（C）基因片段，克隆入真核表达质粒pLNCX2，经测序正确后以脂质体转染PT-67细胞，挑取单克隆并扩增，收集上清感染MN9D细胞，分别用Real Time PCR检测细胞基因表达水平，免疫组织化学染色检测蛋白表达，激光扫描共焦显微镜检测蛋白与线粒体共定位，流式细胞术检测细胞状态及线粒体膜电位状态。结果成功构建了pLNCX2/N、pLNCX2/NAC及pLNCX2/C基因片段的重组真核表达质粒，获得了可稳定表达α-synuclein各基因片段的MN9D细胞株。通过激光扫描共焦显微镜观察，可见α-synuclein/N端与线粒体存在共定位关系；α-synuclein/NAC主要在核内聚集表达；α-synuclein/C端在胞质和胞核内均有表达。JC1染色流式细胞术检测显示，过表达α-synuclein/N实验组细胞线粒体膜电位降低。结论α-synuclein的N端可能定位于线粒体，并参与调节线粒体功能；α-synuclein/NAC虽具有疏水性但却能穿过核膜，聚集在核仁周围；α-synuclein的C端定位于胞质和核内可能参与多种细胞功能。
