To detect retrospectively the phenotype and stability of the E-protein gene in Japanese Encephalitis (JE) virus strain SA14-14-2 used in the live-attenuated JE vaccine prepared at different years, the viral titer was ...To detect retrospectively the phenotype and stability of the E-protein gene in Japanese Encephalitis (JE) virus strain SA14-14-2 used in the live-attenuated JE vaccine prepared at different years, the viral titer was titrated by plaque formation in BHK-21 cell cultures, and the neuro-virulence of viruses was assayed in mice with body weight of 12-14 g by intracerebral inoculation. Meanwhile, the total RNA of virus gene was extracted and amplified by RT-PCR with the designed primers, and then it was purified and cloned to the expression vector pGEM-T. The recombinant plasmid was purified and sequenced. It was found that the loss of viral titer of vaccines stored in -20℃ for longer than 10 years was less than 0.5 Lg PFU/ml. No mice inoculated intracerebrally showed signs of illness or even death. The size of plagues of the vaccine virus remained to be small, and the E genes of primary virus seed SA14-14-2 and the vaccines prepared at different years (1987-2001) were unchanged, including the 8 critical amino acid sites which were different from the parent wild virus strain SA14 and the related neuro-virulence. These results indicate that the genotypic and biological characteristics of the attenuated JE virus strain SA14-14-2 and its vaccines prepared are quite stable without any reversion noted.展开更多
目的探讨基质金属蛋白酶(matrix metalloproteinase,MMP)-2、14及基质金属蛋白酶组织抑制因子-2(tissueinhibitor of metalloproteinase,TIMP-2)在胃癌中的表达及其意义。方法用免疫组织化学方法(SP法)检测80例胃癌手术患者胃癌组织及3...目的探讨基质金属蛋白酶(matrix metalloproteinase,MMP)-2、14及基质金属蛋白酶组织抑制因子-2(tissueinhibitor of metalloproteinase,TIMP-2)在胃癌中的表达及其意义。方法用免疫组织化学方法(SP法)检测80例胃癌手术患者胃癌组织及30例同期癌旁≥5cm正常组织中MMP-2、MMP-14及TIMP-2的表达。结果①在胃癌、正常胃组织中,MMP-2阳性表达率分别为75.00%和36.67%,MMP-14阳性表达率分别为82.50%和33.33%,差异有统计学意义(均P<0.01);TIMP-2阳性表达率分别为45.00%和40.00%,差异无统计学意义。②癌组织中,MMP-2、MMP-14的表达与肿瘤分化程度、浸润深度、淋巴结有无转移和TNM临床分期密切相关(均P<0.01)。TIMP-2的表达仅与淋巴结有无转移相关(P<0.05)。③生存期<2年患者的MMP-2、MMP-14表达率明显高于生存期≥2年患者(均P<0.01)。结论胃癌组织中,MMP-2、MMP-14高表达,联合检测MMP-2、MMP-14或MMP-2、MMP-14、TIMP-2可作为胃癌恶性程度及预后判断的指标。展开更多
文摘To detect retrospectively the phenotype and stability of the E-protein gene in Japanese Encephalitis (JE) virus strain SA14-14-2 used in the live-attenuated JE vaccine prepared at different years, the viral titer was titrated by plaque formation in BHK-21 cell cultures, and the neuro-virulence of viruses was assayed in mice with body weight of 12-14 g by intracerebral inoculation. Meanwhile, the total RNA of virus gene was extracted and amplified by RT-PCR with the designed primers, and then it was purified and cloned to the expression vector pGEM-T. The recombinant plasmid was purified and sequenced. It was found that the loss of viral titer of vaccines stored in -20℃ for longer than 10 years was less than 0.5 Lg PFU/ml. No mice inoculated intracerebrally showed signs of illness or even death. The size of plagues of the vaccine virus remained to be small, and the E genes of primary virus seed SA14-14-2 and the vaccines prepared at different years (1987-2001) were unchanged, including the 8 critical amino acid sites which were different from the parent wild virus strain SA14 and the related neuro-virulence. These results indicate that the genotypic and biological characteristics of the attenuated JE virus strain SA14-14-2 and its vaccines prepared are quite stable without any reversion noted.
文摘目的探讨基质金属蛋白酶(matrix metalloproteinase,MMP)-2、14及基质金属蛋白酶组织抑制因子-2(tissueinhibitor of metalloproteinase,TIMP-2)在胃癌中的表达及其意义。方法用免疫组织化学方法(SP法)检测80例胃癌手术患者胃癌组织及30例同期癌旁≥5cm正常组织中MMP-2、MMP-14及TIMP-2的表达。结果①在胃癌、正常胃组织中,MMP-2阳性表达率分别为75.00%和36.67%,MMP-14阳性表达率分别为82.50%和33.33%,差异有统计学意义(均P<0.01);TIMP-2阳性表达率分别为45.00%和40.00%,差异无统计学意义。②癌组织中,MMP-2、MMP-14的表达与肿瘤分化程度、浸润深度、淋巴结有无转移和TNM临床分期密切相关(均P<0.01)。TIMP-2的表达仅与淋巴结有无转移相关(P<0.05)。③生存期<2年患者的MMP-2、MMP-14表达率明显高于生存期≥2年患者(均P<0.01)。结论胃癌组织中,MMP-2、MMP-14高表达,联合检测MMP-2、MMP-14或MMP-2、MMP-14、TIMP-2可作为胃癌恶性程度及预后判断的指标。
基金supported by grants from The National Natural Scientific Foundation of China(30901577)The Hengyang Technological Bureau of China(2009KJ14)+1 种基金Ministry of Education of Returned Overseas Students to Start Research and Fund Projects(20091590)Aid Program for Science and Technology Innovative Research Team in Higher Educational Institutions of Hunan Province~~
