表观遗传修饰参与了药物成瘾的形成过程,而在药物成瘾过程中组蛋白泛素化水平的变化仍未可知。药物成瘾过程中常表现为多巴胺(dopamine, DA)表达量的升高,因此本研究欲探讨多巴胺升高对神经细胞组蛋白泛素化的影响及其机制。Western印...表观遗传修饰参与了药物成瘾的形成过程,而在药物成瘾过程中组蛋白泛素化水平的变化仍未可知。药物成瘾过程中常表现为多巴胺(dopamine, DA)表达量的升高,因此本研究欲探讨多巴胺升高对神经细胞组蛋白泛素化的影响及其机制。Western印迹结果显示,在终浓度0.8 mmol/L的多巴胺作用8 h后,人神经母细胞瘤细胞系SH-SY5Y细胞中环指蛋白20(ring finger protein 20, RNF20)表达量降低(0.29±0.032 vs. 1.0±0.025,P<0.0001),泛素化组蛋白H2B(H2Bub1)表达量下降(0.28±0.032 vs. 1.0±0.017,P<0.0001)。但是RT-PCR结果显示,多巴胺处理SH-SY5Y细胞后,RNF20在mRNA水平的表达无明显变化。在SH-SY5Y细胞中沉默RNF20的表达,H2Bub1在蛋白质水平的表达明显降低(0.20±0.069 vs. 1.0±0.060,P=0.001)。在加入多巴胺的基础上,分别加入蛋白酶体抑制剂MG132、自噬体形成抑制剂3-MA以及空泡型H^+-ATP酶特异性抑制剂Baf-A1等药物来检测RNF20的降解途径,结果发现,加入MG132、3-MA以及Baf-A1后,RNF20表达量均比DA处理组显著上升(1.51±0.095,P=0.0003; 0.89±0.075,P=0.0021; 2.74±0.099,P<0.0001;vs. 0.27±0.044)。上述结果表明,在SH-SY5Y细胞中,RNF20对H2Bub1具有调控作用,多巴胺可通过泛素化及自噬两种途径促进RNF20降解,从而抑制组蛋白H2B泛素化。展开更多
HIV is a neurotropic virus and can be engulfed macrophagocytes. It invades the brain tissue via the blood-brain barrier and multiplies in the cerebrospinal fluid （CSF）. HIV has been isolated from the brain, spinal m...HIV is a neurotropic virus and can be engulfed macrophagocytes. It invades the brain tissue via the blood-brain barrier and multiplies in the cerebrospinal fluid （CSF）. HIV has been isolated from the brain, spinal marrow, peripheral nerve, and the CSF. Electronic microscopy, immunohistochemistry, and in situ hybridization have demonstrated monocytes and multinucleated cells containing HIV virus.展开更多
Background The global outbreak of influenza A （H1N1） has led to the Ministry of Health of China listing it as one of the A-class infectious diseases. Pneumonia is the most serious complication of influenza A, common...Background The global outbreak of influenza A （H1N1） has led to the Ministry of Health of China listing it as one of the A-class infectious diseases. Pneumonia is the most serious complication of influenza A, commonly causing death. Populations are ordinarily susceptible to influenza A. This study aimed to investigate the imaging manifestation features of critical influenza A （H1 N1 ） pneumonia and to improve its diagnostic techniques. Methods A total of seven death cases from critical influenza A （H1N1） pneumonia were retrospectively analyzed on their imaging manifestations and autopsy data. Pulmonary CT scanning was performed for five cases, with one receiving additional chest X-ray and chest CT scanning, and chest postero-anterior position X-ray examination was performed for other two. Autopsy was performed for five cases and postmortem examinations were performed for other two cases. Results The seven cases of influenza A showed critical manifestations in 4-7 days after symptoms onset, with two having basic diseases of diabetes and one being pregnant. Extensive blurry high-density shadows of bilateral lungs were found in three cases, which were most obvious in middle and inferior parts of lungs. Pulmonary CT scanning revealed bilateral flaky parenchymal shadows in peripheral, dorsal and fundus segments of the middle-inferior parts of lungs, with one case of complicated pneumothorax, atelectasis and pleural effusion and another case of thin-walled cavity and dilated bronchi shadows in the superior parts of lungs. Conclusions Diagnostic imaging is an important assessing tool for critical influenza A （H1N1） pneumonia. The imaging manifestations are characteristic instead of being specific. The definitive diagnosis can be made in combination with clinical examinations and laboratory tests.展开更多
目的在体内,热休克蛋白90α(heat shock protein 90α,HSP90α)对肿瘤的发生发展起着重要的作用。HSP90α抑制剂在体内可阻断肿瘤赖以生存的信号通路网络。本研究探讨HSP90α特异性抑制剂烯丙胺基-17-去甲氧基格尔德(17-allyl-amino-17-...目的在体内,热休克蛋白90α(heat shock protein 90α,HSP90α)对肿瘤的发生发展起着重要的作用。HSP90α抑制剂在体内可阻断肿瘤赖以生存的信号通路网络。本研究探讨HSP90α特异性抑制剂烯丙胺基-17-去甲氧基格尔德(17-allyl-amino-17-demethoxygeldanamycin,17-AAG)对甲状腺癌细胞增殖和迁移的影响及相关信号分子的调控机制。方法体外培养甲状腺乳头状癌细胞TPC-1和甲状腺未分化癌细胞TAK,分别给予0.001～10.000μmol/L浓度的17-AAG作用24和48h。MTT实验、划痕实验观察细胞生长和迁移情况。蛋白质印迹法检测细胞HSP90α及信号传导及转录激活因子(signal transducers and activators of transcription,STAT3)、丝氨酸/苏氨酸蛋白激酶(serine/threonine protein kinase,AKT)、细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)相关信号蛋白的表达及磷酸化水平变化。结果与未处理的细胞相比,经0.001～10.000μmol/L浓度的17-AAG作用24和48h的TPC-1和TAK细胞增殖能力降低,且有明显的时间和剂量依赖性。TPC-1(F=7.326,P=0.005)和TAK细胞(F=22.770,P<0.001)浓度组间比较差异有统计学意义,时间组间比较差异有统计学意义,F值分别为1 048.000和663.800,均P<0.001。与未处理的细胞相比,经0.1和1.0μmol/L浓度的17-AAG作用24h的TPC-1和TAK细胞迁移能力降低,各浓度组间比较差异有统计学意义,F=1.228,P<0.005;两细胞株间比较差异无统计学意义,F=0.365,P=0.578。经0.1和1.0μmol/L浓度的17-AAG作用24h的TPC-1和TAK细胞以及STAT3、AKT、ERK蛋白磷酸化水平降低,却对STAT3和ERK总蛋白表达影响差异无统计学意义,P>0.05。结论 17-AAG通过下调甲状腺癌细胞HSP90α相关信号蛋白STAT3/AKT/ERK磷酸化水平,从而抑制甲状腺癌细胞的增殖和迁移能力。展开更多
文摘表观遗传修饰参与了药物成瘾的形成过程,而在药物成瘾过程中组蛋白泛素化水平的变化仍未可知。药物成瘾过程中常表现为多巴胺(dopamine, DA)表达量的升高,因此本研究欲探讨多巴胺升高对神经细胞组蛋白泛素化的影响及其机制。Western印迹结果显示,在终浓度0.8 mmol/L的多巴胺作用8 h后,人神经母细胞瘤细胞系SH-SY5Y细胞中环指蛋白20(ring finger protein 20, RNF20)表达量降低(0.29±0.032 vs. 1.0±0.025,P<0.0001),泛素化组蛋白H2B(H2Bub1)表达量下降(0.28±0.032 vs. 1.0±0.017,P<0.0001)。但是RT-PCR结果显示,多巴胺处理SH-SY5Y细胞后,RNF20在mRNA水平的表达无明显变化。在SH-SY5Y细胞中沉默RNF20的表达,H2Bub1在蛋白质水平的表达明显降低(0.20±0.069 vs. 1.0±0.060,P=0.001)。在加入多巴胺的基础上,分别加入蛋白酶体抑制剂MG132、自噬体形成抑制剂3-MA以及空泡型H^+-ATP酶特异性抑制剂Baf-A1等药物来检测RNF20的降解途径,结果发现,加入MG132、3-MA以及Baf-A1后,RNF20表达量均比DA处理组显著上升(1.51±0.095,P=0.0003; 0.89±0.075,P=0.0021; 2.74±0.099,P<0.0001;vs. 0.27±0.044)。上述结果表明,在SH-SY5Y细胞中,RNF20对H2Bub1具有调控作用,多巴胺可通过泛素化及自噬两种途径促进RNF20降解,从而抑制组蛋白H2B泛素化。
基金This work was supported by special funds of the National Serious Infectious Diseases Foundation of China （No. 2008ZX10001-006）, grants from Beijing Natural Science Foundation （No. 7132108）, and the Academic Leader of 215 High-level Health Personnel, Beijing, China （No. 2009-3-55）.
文摘HIV is a neurotropic virus and can be engulfed macrophagocytes. It invades the brain tissue via the blood-brain barrier and multiplies in the cerebrospinal fluid （CSF）. HIV has been isolated from the brain, spinal marrow, peripheral nerve, and the CSF. Electronic microscopy, immunohistochemistry, and in situ hybridization have demonstrated monocytes and multinucleated cells containing HIV virus.
文摘Background The global outbreak of influenza A （H1N1） has led to the Ministry of Health of China listing it as one of the A-class infectious diseases. Pneumonia is the most serious complication of influenza A, commonly causing death. Populations are ordinarily susceptible to influenza A. This study aimed to investigate the imaging manifestation features of critical influenza A （H1 N1 ） pneumonia and to improve its diagnostic techniques. Methods A total of seven death cases from critical influenza A （H1N1） pneumonia were retrospectively analyzed on their imaging manifestations and autopsy data. Pulmonary CT scanning was performed for five cases, with one receiving additional chest X-ray and chest CT scanning, and chest postero-anterior position X-ray examination was performed for other two. Autopsy was performed for five cases and postmortem examinations were performed for other two cases. Results The seven cases of influenza A showed critical manifestations in 4-7 days after symptoms onset, with two having basic diseases of diabetes and one being pregnant. Extensive blurry high-density shadows of bilateral lungs were found in three cases, which were most obvious in middle and inferior parts of lungs. Pulmonary CT scanning revealed bilateral flaky parenchymal shadows in peripheral, dorsal and fundus segments of the middle-inferior parts of lungs, with one case of complicated pneumothorax, atelectasis and pleural effusion and another case of thin-walled cavity and dilated bronchi shadows in the superior parts of lungs. Conclusions Diagnostic imaging is an important assessing tool for critical influenza A （H1N1） pneumonia. The imaging manifestations are characteristic instead of being specific. The definitive diagnosis can be made in combination with clinical examinations and laboratory tests.
文摘目的在体内,热休克蛋白90α(heat shock protein 90α,HSP90α)对肿瘤的发生发展起着重要的作用。HSP90α抑制剂在体内可阻断肿瘤赖以生存的信号通路网络。本研究探讨HSP90α特异性抑制剂烯丙胺基-17-去甲氧基格尔德(17-allyl-amino-17-demethoxygeldanamycin,17-AAG)对甲状腺癌细胞增殖和迁移的影响及相关信号分子的调控机制。方法体外培养甲状腺乳头状癌细胞TPC-1和甲状腺未分化癌细胞TAK,分别给予0.001～10.000μmol/L浓度的17-AAG作用24和48h。MTT实验、划痕实验观察细胞生长和迁移情况。蛋白质印迹法检测细胞HSP90α及信号传导及转录激活因子(signal transducers and activators of transcription,STAT3)、丝氨酸/苏氨酸蛋白激酶(serine/threonine protein kinase,AKT)、细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)相关信号蛋白的表达及磷酸化水平变化。结果与未处理的细胞相比,经0.001～10.000μmol/L浓度的17-AAG作用24和48h的TPC-1和TAK细胞增殖能力降低,且有明显的时间和剂量依赖性。TPC-1(F=7.326,P=0.005)和TAK细胞(F=22.770,P<0.001)浓度组间比较差异有统计学意义,时间组间比较差异有统计学意义,F值分别为1 048.000和663.800,均P<0.001。与未处理的细胞相比,经0.1和1.0μmol/L浓度的17-AAG作用24h的TPC-1和TAK细胞迁移能力降低,各浓度组间比较差异有统计学意义,F=1.228,P<0.005;两细胞株间比较差异无统计学意义,F=0.365,P=0.578。经0.1和1.0μmol/L浓度的17-AAG作用24h的TPC-1和TAK细胞以及STAT3、AKT、ERK蛋白磷酸化水平降低,却对STAT3和ERK总蛋白表达影响差异无统计学意义,P>0.05。结论 17-AAG通过下调甲状腺癌细胞HSP90α相关信号蛋白STAT3/AKT/ERK磷酸化水平,从而抑制甲状腺癌细胞的增殖和迁移能力。
