目的检测空泡蛋白分类相关蛋白33A(vacuolar protein sorting-associated protein 33A,VPS33A)在老年肺结核患者外周血中的表达情况,并探讨其与临床预后的相关性。方法收集麻城市人民医院2019年2月~2021年2月收治的95例老年肺结核患者...目的检测空泡蛋白分类相关蛋白33A(vacuolar protein sorting-associated protein 33A,VPS33A)在老年肺结核患者外周血中的表达情况,并探讨其与临床预后的相关性。方法收集麻城市人民医院2019年2月~2021年2月收治的95例老年肺结核患者为研究组,另外选取同期健康体检者90例为对照组。采用实时荧光定量PCR(qRT-PCR)检测外周血中VPS33A,Toll样受体2(TLR2)表达情况,酶联免疫吸附法(ELISA)检测外周血中白细胞介素-6(IL-6)水平。肺结核患者给予标准化抗结核方案治疗(2HRZE/4HR),治疗后随访12个月,根据患者预后将其分为预后良好组(n=58)和预后不良组(n=37),采用受试者工作特征(ROC)曲线分析VPS33A对患者预后不良的预测价值;Pearson法分析预后不良组血清VPS33A与IL-6,TLR2的相关性;Logistic回归分析老年肺结核患者预后不良的影响因素。结果与对照组比较,治疗前研究组外周血VPS33A(0.87±0.22vs1.02±0.23)水平明显降低,IL-6(22.13±5.36 pg/mlvs18.53±4.68pg/ml),TLR2(1.26±0.25vs0.99±0.21)水平明显升高,差异具有统计学意义(t=6.170,6.682,11.241,均P<0.05);与治疗前研究组比较,治疗后研究组外周血VPS33A(0.95±0.25)水平明显升高,IL-6(19.66±5.44 pg/ml),TLR2(1.11±0.2)水平明显降低,差异具有统计学意义(t-3.336,4.648,6.331,均P<0.05)。预后不良组外周血VPS33A水平(0.71±0.16)显著低于预后良好组(0.97±0.22);IL-6(26.41±7.16pg/ml),TLR2(1.35±0.26)水平显著高于预后良好组(19.40±4.21pg/ml,1.20±0.24),差异具有统计学意义(t=5.555,6.012,2.875,均P<0.05)。经Pearson法分析发现,治疗前外周血VPS33A分别与IL-6,TLR2水平呈负相关(r=-0.649,-0.634,均P<0.05)。ROC曲线分析结果显示VPS33A,IL-6和TLR2预测老年肺结核预后的AUC分别为0.805,0.842和0.882,三者联合AUC为0.971,明显高于三者单独预测(Z=4.010,3.136,2.285,P=0.001,0.002,0.022);Logistic回归分析显示外周血VPS33A,IL-6和TLR2是老年肺结核患者预后不良的影响因素(均P<0.05)。结论VPS33A在老年肺结核患者外周血中的表达下调,对老年肺结核患者预后有较好的预测价值,且是预后不良的影响因素。展开更多
PC12 cell injury was induced using 20 μM amyloid β-protein 25-35 to establish a model of Alzheimer＇s disease. The cells were then treated with 5, 10, and 25 μM Schisandrin B. Methylthiazolyldiphenyl-tetrazolium br...PC12 cell injury was induced using 20 μM amyloid β-protein 25-35 to establish a model of Alzheimer＇s disease. The cells were then treated with 5, 10, and 25 μM Schisandrin B. Methylthiazolyldiphenyl-tetrazolium bromide assays and Hoechst 33342 staining results showed that with increasing Schisandrin B concentration, the survival rate of PC12 cells injured by amyloid β-protein 25-35 gradually increased and the rate of apoptosis gradually decreased. Reverse transcription-PCR, immunocytochemical staining and western blot results showed that with increasing Schisandrin B concentration, the mRNA and protein expression of vacuolar protein sorting 35 and amyloid precursor protein were gradually decreased. Vacuolar protein sorting 35 and amyloid precursor protein showed a consistent trend for change. These findings suggest that 5, 10, and 25 μM Schisandrin B antagonizes the cellular injury induced by amyloid β-protein 25-35 in a dose-dependent manner. This may be caused by decreasing the expression of vacuolar protein sorting 35 and amyloid precursor protein.展开更多
This study examined the anti-hepatitis B virus （HBV） effect of wild-type （WT） vacuolar protein sorting 4B （VPS4B） and its dominant negative （DN） mutant VPS4B-K180Q in vivo in order to further explore the relat...This study examined the anti-hepatitis B virus （HBV） effect of wild-type （WT） vacuolar protein sorting 4B （VPS4B） and its dominant negative （DN） mutant VPS4B-K180Q in vivo in order to further explore the relationship between HBV and the host cellular factor VPS4. VPS4B gene was amplified from Huh7 cells by RT-PCR and cloned into the eukaryotic expression vector pXF3H. Then, the VPS4B plasmid and the VPS4B-K180Q mutation plasmid were constructed by using the overlap extension PCR site-directed mutagenesis technique. VPS4B and HBV vectors were co-delivered into mice by the hydrodynamic tail-vein injection to establish HBV vector-based models. Quantities of HBsAg and HBeAg in the mouse sera were determined by ElectroChemiLuminescence （ECL）. HBV DNA in sera was measured by real-time quantitative PCR. Southern blot analysis was used to assay the intracellular HBV nuclear capsid-related DNA, real-time quantitative PCR to detect the HBV-related mRNA and immunohistochemical staining to observe the HBcAg expression in the mouse liver tissues. Our results showed that VPS4B and its mutant VPS4B-K180Q could decrease the levels of serum HBsAg, HBeAg and HBV-DNA. In addition, the HBV DNA replication and the mRNA level of HBV in the liver tissues of treated mice could be suppressed by VPS4B and VPS4B-K180Q. It was also found that VPS4B and VPS4B-K180Q had an ability to inhibit core antigen expression in the infected mouse liver. Furthermore, the anti-HBV effect of mutant VPS4B-K180Q was more potent than that of wild-type VPS4B. Taken together, it was concluded that VPS4B and its DN mutant VPS4B-K180Q have anti-HBV effect in vivo, which helps develop molecular therapeutic strategies for HBV infection.展开更多
文摘目的检测空泡蛋白分类相关蛋白33A(vacuolar protein sorting-associated protein 33A,VPS33A)在老年肺结核患者外周血中的表达情况,并探讨其与临床预后的相关性。方法收集麻城市人民医院2019年2月~2021年2月收治的95例老年肺结核患者为研究组,另外选取同期健康体检者90例为对照组。采用实时荧光定量PCR(qRT-PCR)检测外周血中VPS33A,Toll样受体2(TLR2)表达情况,酶联免疫吸附法(ELISA)检测外周血中白细胞介素-6(IL-6)水平。肺结核患者给予标准化抗结核方案治疗(2HRZE/4HR),治疗后随访12个月,根据患者预后将其分为预后良好组(n=58)和预后不良组(n=37),采用受试者工作特征(ROC)曲线分析VPS33A对患者预后不良的预测价值;Pearson法分析预后不良组血清VPS33A与IL-6,TLR2的相关性;Logistic回归分析老年肺结核患者预后不良的影响因素。结果与对照组比较,治疗前研究组外周血VPS33A(0.87±0.22vs1.02±0.23)水平明显降低,IL-6(22.13±5.36 pg/mlvs18.53±4.68pg/ml),TLR2(1.26±0.25vs0.99±0.21)水平明显升高,差异具有统计学意义(t=6.170,6.682,11.241,均P<0.05);与治疗前研究组比较,治疗后研究组外周血VPS33A(0.95±0.25)水平明显升高,IL-6(19.66±5.44 pg/ml),TLR2(1.11±0.2)水平明显降低,差异具有统计学意义(t-3.336,4.648,6.331,均P<0.05)。预后不良组外周血VPS33A水平(0.71±0.16)显著低于预后良好组(0.97±0.22);IL-6(26.41±7.16pg/ml),TLR2(1.35±0.26)水平显著高于预后良好组(19.40±4.21pg/ml,1.20±0.24),差异具有统计学意义(t=5.555,6.012,2.875,均P<0.05)。经Pearson法分析发现,治疗前外周血VPS33A分别与IL-6,TLR2水平呈负相关(r=-0.649,-0.634,均P<0.05)。ROC曲线分析结果显示VPS33A,IL-6和TLR2预测老年肺结核预后的AUC分别为0.805,0.842和0.882,三者联合AUC为0.971,明显高于三者单独预测(Z=4.010,3.136,2.285,P=0.001,0.002,0.022);Logistic回归分析显示外周血VPS33A,IL-6和TLR2是老年肺结核患者预后不良的影响因素(均P<0.05)。结论VPS33A在老年肺结核患者外周血中的表达下调,对老年肺结核患者预后有较好的预测价值,且是预后不良的影响因素。
基金supported by the National 985 Project "linguistic science technology and the construction of interdisciplinary innovation platform in current society",No.985yk002the National 985 Project "cognitive and neural information science platform",No.904273258
文摘PC12 cell injury was induced using 20 μM amyloid β-protein 25-35 to establish a model of Alzheimer＇s disease. The cells were then treated with 5, 10, and 25 μM Schisandrin B. Methylthiazolyldiphenyl-tetrazolium bromide assays and Hoechst 33342 staining results showed that with increasing Schisandrin B concentration, the survival rate of PC12 cells injured by amyloid β-protein 25-35 gradually increased and the rate of apoptosis gradually decreased. Reverse transcription-PCR, immunocytochemical staining and western blot results showed that with increasing Schisandrin B concentration, the mRNA and protein expression of vacuolar protein sorting 35 and amyloid precursor protein were gradually decreased. Vacuolar protein sorting 35 and amyloid precursor protein showed a consistent trend for change. These findings suggest that 5, 10, and 25 μM Schisandrin B antagonizes the cellular injury induced by amyloid β-protein 25-35 in a dose-dependent manner. This may be caused by decreasing the expression of vacuolar protein sorting 35 and amyloid precursor protein.
基金supported by a grant from the National Mega Research Program of China(No.2008ZX10002-011)
文摘This study examined the anti-hepatitis B virus （HBV） effect of wild-type （WT） vacuolar protein sorting 4B （VPS4B） and its dominant negative （DN） mutant VPS4B-K180Q in vivo in order to further explore the relationship between HBV and the host cellular factor VPS4. VPS4B gene was amplified from Huh7 cells by RT-PCR and cloned into the eukaryotic expression vector pXF3H. Then, the VPS4B plasmid and the VPS4B-K180Q mutation plasmid were constructed by using the overlap extension PCR site-directed mutagenesis technique. VPS4B and HBV vectors were co-delivered into mice by the hydrodynamic tail-vein injection to establish HBV vector-based models. Quantities of HBsAg and HBeAg in the mouse sera were determined by ElectroChemiLuminescence （ECL）. HBV DNA in sera was measured by real-time quantitative PCR. Southern blot analysis was used to assay the intracellular HBV nuclear capsid-related DNA, real-time quantitative PCR to detect the HBV-related mRNA and immunohistochemical staining to observe the HBcAg expression in the mouse liver tissues. Our results showed that VPS4B and its mutant VPS4B-K180Q could decrease the levels of serum HBsAg, HBeAg and HBV-DNA. In addition, the HBV DNA replication and the mRNA level of HBV in the liver tissues of treated mice could be suppressed by VPS4B and VPS4B-K180Q. It was also found that VPS4B and VPS4B-K180Q had an ability to inhibit core antigen expression in the infected mouse liver. Furthermore, the anti-HBV effect of mutant VPS4B-K180Q was more potent than that of wild-type VPS4B. Taken together, it was concluded that VPS4B and its DN mutant VPS4B-K180Q have anti-HBV effect in vivo, which helps develop molecular therapeutic strategies for HBV infection.
