目的:观察补肾益髓(Bu Shen Yi Sui,BSYS)方及其拆方补肾(Bu Shen,BS)和化痰活血(Hua Tan Huo Xue,HTHX)方对实验性自身免疫性脑脊髓炎(Experimental Autoimmune Encephalomyelitis,EAE)小鼠脑和脊髓中p-Tau和Beta-tubulin的作用。方法:...目的:观察补肾益髓(Bu Shen Yi Sui,BSYS)方及其拆方补肾(Bu Shen,BS)和化痰活血(Hua Tan Huo Xue,HTHX)方对实验性自身免疫性脑脊髓炎(Experimental Autoimmune Encephalomyelitis,EAE)小鼠脑和脊髓中p-Tau和Beta-tubulin的作用。方法:C57BL/6小鼠雌性随机分为正常对照(NC)、模型(MO)、醋酸泼尼松(PA)、梓醇(CA)、BSYS、BS和HTHX组。造模当天与造模后第7天于小鼠背部两侧皮下注射抗原,即50μg髓鞘少突胶质细胞糖蛋白(MOG)35-55、完全弗氏佐剂(CFA)与结核分支杆菌(TB)混合,并在免疫当天和2 d后腹腔注射500 ng的百日咳毒素(PTX)。每天对小鼠进行灌胃,观察其体重及神经功能评分,并于造模第20 d和第40 d分别取脑和脊髓进行病理检测。采用免疫组化法检测p-Tau和beta-tubulin的表达。结果:与MO组比较,BSYS、BS和HTHX组体重与神经功能评分均出现明显上升,HE染色可观察到炎细胞显著减少,核固缩减轻,同时,BSYS、BS和HTHX方均可明显下调p-Tau表达(P<0.05,P<0.01),上调beta-tubulin蛋白表达(P<0.05,P<0.01)。结论:BSYS方及其拆方BS和HTHX均对EAE小鼠有神经保护作用,表现在减少体重丢失,降低神经功能评分,减轻炎炎性细胞浸润,减轻轴突损伤及促进其修复,尤以BSYS全方更为显著。展开更多
Endogenous tubulin promoter has been widely used for expressing foreign genes in green algae, but the efficiency and feasibility of endogenous tubulin promoter in the economically important Porphyra yezoensis (Rhodoph...Endogenous tubulin promoter has been widely used for expressing foreign genes in green algae, but the efficiency and feasibility of endogenous tubulin promoter in the economically important Porphyra yezoensis (Rhodophyta) are unknown. In this study, the flanking sequences of beta-tubulin gene from P. yezoensis were amplified and two transient expression vectors were con-structed to determine their transcription promoting feasibility for foreign gene gusA. The testing vector pATubGUS was constructed by inserting 5’- and 3’-flanking regions (Tub5’ and Tub3’) up- and down-stream of β-glucuronidase (GUS) gene (gusA), respectively, into pA, a derivative of pCAT?3-enhancer vector. The control construct, pAGUSTub3, contains only gusA and Tub3’. These con-structs were electroporated into P. yezoensis protoplasts and the GUS activities were quantitatively analyzed by spectrometry. The results demonstrated that gusA gene was efficiently expressed in P. yezoensis protoplasts under the regulation of 5’-flanking sequence of the beta-tubulin gene. More interestingly, the pATubGUS produced stronger GUS activity in P. yezoensis protoplasts when com-pared to the result from pBI221, in which the gusA gene was directed by a constitutive CaMV 35 S promoter. The data suggest that the integration of P. yezoensis protoplast and its endogenous beta-tubulin flanking sequences is a potential novel system for foreign gene expression.展开更多
Objective: The aim of this study was to investigate the impact of beta-elemene injection on the growth and beta-tubulin of human hepatocarcinoma HepG2 cells. Methods: Cell proliferation was assessed by MTT assay. Cell...Objective: The aim of this study was to investigate the impact of beta-elemene injection on the growth and beta-tubulin of human hepatocarcinoma HepG2 cells. Methods: Cell proliferation was assessed by MTT assay. Cell cycle distribution was detected by flow cytometry(FCM). The mRNA expression of beta-tubulin was measured by RT-PCR. Western blot analysis was used to determine protein expression of beta-tubulin and the polymerization of beta-tubulin. Results: Beta-elemene injection inhibited HepG2 cells proliferation in a dose- and time-dependent manner; FCM analysis indicated beta-elemene injection induced cell cycle arrested at S phase. RT-PCR and western-blot analysis showed that beta-elemene injection down-regulated beta-tubulin expression at both mRNA and protein levels, presenting a dose-dependent manner. Moreover, beta-elemene injection reduced the polymerization of microtubules in a dose-dependent manner. Conclusion: Beta-elemene injection can inhibit the proliferation of hepatoma HepG2 cells, the mechanism might be partly related to the down-regulation of beta-tubulin and inhibition of microtubular polymerization.展开更多
Objective Based on our previous established cohort of myelodysplastic syndrome(MDS),we investigated the potential effect of beta-tubulin(TUBB)gene in the transformation of MDS into acute leukemia.Methods From our nest...Objective Based on our previous established cohort of myelodysplastic syndrome(MDS),we investigated the potential effect of beta-tubulin(TUBB)gene in the transformation of MDS into acute leukemia.Methods From our nested case-control study cohort of MDS patients,we chose 11 paired transformed and non-transformed展开更多
目的:探讨β微管蛋白(β-tubulin)在非小细胞肺癌(NSCLC)中的表达情况,及其与诺维本/铂类(NP)术后辅助化疗预后的关系。方法:回顾性分析广州医学院第一附属医院2002年1月至2009年12月收治的84例手术完全切除或部分切除的NSCLC患者,术后...目的:探讨β微管蛋白(β-tubulin)在非小细胞肺癌(NSCLC)中的表达情况,及其与诺维本/铂类(NP)术后辅助化疗预后的关系。方法:回顾性分析广州医学院第一附属医院2002年1月至2009年12月收治的84例手术完全切除或部分切除的NSCLC患者,术后以NP方案辅助化疗。应用免疫组织化学S-P法检测患者手术标本β-tubulin中蛋白的表达,进一步分析其与无病生存时间(DFS)和总生存时间(OS)的关系。结果:NSCLC患者中β-tubulin蛋白的低表达率为42.9%(36/84),高表达率为57.1%(48/84),男性β-tubulin高表达患者多于低表达患者(68.2%vs.31.8%,P=0.032);Ⅰ～Ⅱ期β-tubulin高表达患者多于低表达患者(70.7%vs.29.3,P=0.014);且行根治性手术β-tubulin高表达患者多于低表达患者(61.6%vs.38.4%,P=0.032),与NSCLC患者的年龄、病理类型、病理分级、淋巴结转移、吸烟史和放疗均无统计学相关性(P>0.05)。单因素生存分析显示,经低表达β-tubulin的NP方案辅助化疗的NSCLC患者,中位DFS短于高表达者(22.6 vs 69.6个月),两者之间差异无统计学意义(P=0.052),但高表达β-tubulin患者的DFS在术后13个月后明显优于低表达者;进一步分层分析显示,年龄>60岁(P=0.032)、腺癌(P=0.034)、中高分化程度(P=0.028)、不吸烟(P=0.016)的β-tubulin高表达者的DFS均明显优于低表达者。、多因素生存分析显示,β-tubulin的表达(RR=2.213,P=0.025)和临床分期(RR=0.319,P<0.0001)可作为影响DFS的独立预后因素;临床分期(RR=0.426,P=0.010)和放疗(RR=2.381,P=0.026)可作为影响OS预后的独立指标。。结论:在术后辅以NP化疗NSCLC患者中,β-tubulin高表达者预后比低表达者好,术后化疗前对β-tubulin蛋白进行免疫组织化学检测,有利于制定个体化治疗方案,可能改善生存。展开更多
文摘目的:观察补肾益髓(Bu Shen Yi Sui,BSYS)方及其拆方补肾(Bu Shen,BS)和化痰活血(Hua Tan Huo Xue,HTHX)方对实验性自身免疫性脑脊髓炎(Experimental Autoimmune Encephalomyelitis,EAE)小鼠脑和脊髓中p-Tau和Beta-tubulin的作用。方法:C57BL/6小鼠雌性随机分为正常对照(NC)、模型(MO)、醋酸泼尼松(PA)、梓醇(CA)、BSYS、BS和HTHX组。造模当天与造模后第7天于小鼠背部两侧皮下注射抗原,即50μg髓鞘少突胶质细胞糖蛋白(MOG)35-55、完全弗氏佐剂(CFA)与结核分支杆菌(TB)混合,并在免疫当天和2 d后腹腔注射500 ng的百日咳毒素(PTX)。每天对小鼠进行灌胃,观察其体重及神经功能评分,并于造模第20 d和第40 d分别取脑和脊髓进行病理检测。采用免疫组化法检测p-Tau和beta-tubulin的表达。结果:与MO组比较,BSYS、BS和HTHX组体重与神经功能评分均出现明显上升,HE染色可观察到炎细胞显著减少,核固缩减轻,同时,BSYS、BS和HTHX方均可明显下调p-Tau表达(P<0.05,P<0.01),上调beta-tubulin蛋白表达(P<0.05,P<0.01)。结论:BSYS方及其拆方BS和HTHX均对EAE小鼠有神经保护作用,表现在减少体重丢失,降低神经功能评分,减轻炎炎性细胞浸润,减轻轴突损伤及促进其修复,尤以BSYS全方更为显著。
文摘Endogenous tubulin promoter has been widely used for expressing foreign genes in green algae, but the efficiency and feasibility of endogenous tubulin promoter in the economically important Porphyra yezoensis (Rhodophyta) are unknown. In this study, the flanking sequences of beta-tubulin gene from P. yezoensis were amplified and two transient expression vectors were con-structed to determine their transcription promoting feasibility for foreign gene gusA. The testing vector pATubGUS was constructed by inserting 5’- and 3’-flanking regions (Tub5’ and Tub3’) up- and down-stream of β-glucuronidase (GUS) gene (gusA), respectively, into pA, a derivative of pCAT?3-enhancer vector. The control construct, pAGUSTub3, contains only gusA and Tub3’. These con-structs were electroporated into P. yezoensis protoplasts and the GUS activities were quantitatively analyzed by spectrometry. The results demonstrated that gusA gene was efficiently expressed in P. yezoensis protoplasts under the regulation of 5’-flanking sequence of the beta-tubulin gene. More interestingly, the pATubGUS produced stronger GUS activity in P. yezoensis protoplasts when com-pared to the result from pBI221, in which the gusA gene was directed by a constitutive CaMV 35 S promoter. The data suggest that the integration of P. yezoensis protoplast and its endogenous beta-tubulin flanking sequences is a potential novel system for foreign gene expression.
基金Supported by grants from the National Natural Science Foundation of China(No.81173615)the Scientific Research Foundation for the Returned Overseas Chinese Scholars and State Education Ministrythe Specialized Research Fund for the Doctoral Program of Higher Education(No.20102105120002)
文摘Objective: The aim of this study was to investigate the impact of beta-elemene injection on the growth and beta-tubulin of human hepatocarcinoma HepG2 cells. Methods: Cell proliferation was assessed by MTT assay. Cell cycle distribution was detected by flow cytometry(FCM). The mRNA expression of beta-tubulin was measured by RT-PCR. Western blot analysis was used to determine protein expression of beta-tubulin and the polymerization of beta-tubulin. Results: Beta-elemene injection inhibited HepG2 cells proliferation in a dose- and time-dependent manner; FCM analysis indicated beta-elemene injection induced cell cycle arrested at S phase. RT-PCR and western-blot analysis showed that beta-elemene injection down-regulated beta-tubulin expression at both mRNA and protein levels, presenting a dose-dependent manner. Moreover, beta-elemene injection reduced the polymerization of microtubules in a dose-dependent manner. Conclusion: Beta-elemene injection can inhibit the proliferation of hepatoma HepG2 cells, the mechanism might be partly related to the down-regulation of beta-tubulin and inhibition of microtubular polymerization.
文摘Objective Based on our previous established cohort of myelodysplastic syndrome(MDS),we investigated the potential effect of beta-tubulin(TUBB)gene in the transformation of MDS into acute leukemia.Methods From our nested case-control study cohort of MDS patients,we chose 11 paired transformed and non-transformed
文摘目的:探讨β微管蛋白(β-tubulin)在非小细胞肺癌(NSCLC)中的表达情况,及其与诺维本/铂类(NP)术后辅助化疗预后的关系。方法:回顾性分析广州医学院第一附属医院2002年1月至2009年12月收治的84例手术完全切除或部分切除的NSCLC患者,术后以NP方案辅助化疗。应用免疫组织化学S-P法检测患者手术标本β-tubulin中蛋白的表达,进一步分析其与无病生存时间(DFS)和总生存时间(OS)的关系。结果:NSCLC患者中β-tubulin蛋白的低表达率为42.9%(36/84),高表达率为57.1%(48/84),男性β-tubulin高表达患者多于低表达患者(68.2%vs.31.8%,P=0.032);Ⅰ～Ⅱ期β-tubulin高表达患者多于低表达患者(70.7%vs.29.3,P=0.014);且行根治性手术β-tubulin高表达患者多于低表达患者(61.6%vs.38.4%,P=0.032),与NSCLC患者的年龄、病理类型、病理分级、淋巴结转移、吸烟史和放疗均无统计学相关性(P>0.05)。单因素生存分析显示,经低表达β-tubulin的NP方案辅助化疗的NSCLC患者,中位DFS短于高表达者(22.6 vs 69.6个月),两者之间差异无统计学意义(P=0.052),但高表达β-tubulin患者的DFS在术后13个月后明显优于低表达者;进一步分层分析显示,年龄>60岁(P=0.032)、腺癌(P=0.034)、中高分化程度(P=0.028)、不吸烟(P=0.016)的β-tubulin高表达者的DFS均明显优于低表达者。、多因素生存分析显示,β-tubulin的表达(RR=2.213,P=0.025)和临床分期(RR=0.319,P<0.0001)可作为影响DFS的独立预后因素;临床分期(RR=0.426,P=0.010)和放疗(RR=2.381,P=0.026)可作为影响OS预后的独立指标。。结论:在术后辅以NP化疗NSCLC患者中,β-tubulin高表达者预后比低表达者好,术后化疗前对β-tubulin蛋白进行免疫组织化学检测,有利于制定个体化治疗方案,可能改善生存。
