人白介素6受体基因在痘苗病毒载体系统中的表达

人ＩＬ－６受体是一个在各种细胞上广泛表达的跨膜糖蛋白分子,是ＩＬ－６发挥细胞效应所必需的。本文通过将ＩＬ－６ＲｃＤＮＡ重组到痘苗病毒的ＴＫ基因中构建成重组痘苗病毒ＶＩＬ６Ｒ。细胞原位杂交和ＡＰＡＡＰ染色结果表明,感染ＶＩＬ６Ｒ后的Ｖｅｒｏ细胞中,ＩＬ－６Ｒ在ｍＲＮＡ和蛋白水平上均呈现较强的表达。Ｗｅｓｔｅｒｎｂｌｏｔ分析所表达的分子量为８０ｋＤ,表明所表达的产物是糖基化的。ＩＬ－６结合试验表明,表达的膜ＩＬ－６Ｒ能够结合ｒＩＬ－６,说明它是有功能的。利用ＶＩＬ６Ｒ免疫小鼠后,能够刺激较强的抗体产生。从而为进一步研究ＩＬ－６Ｒ的信号传导和构效关系提供了基础。     

木薯果胶甲基酯酶抑制因子MePMEI1的生物信息学分析

为探讨木薯MePMEI1的分子结构特征。通过PCR扩增和测序技术及生物信息学分析工具对木薯MePMEI1基因进行克隆、测序及相关生物信息学分析。结果表明木薯MePMEI1基因编码区全长609 bp,编码202个氨基酸残基;MePMEI1基因编码蛋白分子量21.78 k D,理论等电点(pI)约为5.51;生物信息学预测发现,木薯MePMEI1蛋白是稳定的亲水蛋白;具有跨膜区为分泌蛋白;含有1个PMEI结构域,1个糖基化位点,31个磷酸化位点;二、三级结构以α螺旋和无规则卷曲为主。该蛋白的生物功能可能与细胞被膜、酶和生长因子等相关。木薯MePMEI1基因的生物信息学分析为进一步研究其遗传特性和生理生化机制提供了理论依据。     

Vaccination of chickens with DNA vaccine encoding Eimeria acervulina 3-1E and chicken IL-15 offers protection against homologous challenge

Eimeria acervulina 3-1E antigen gene and mature chicken interleukin 15 (mChIL-15) gene were cloned into expression vector pcDNA3.1(+) in different forms, produced DNA vaccine pcDNA3.1-3-1E, and pcDNA3.1-3-1E-linker-mChIL-15 co-expressing E. acervulina 3-1E gene and mChIL-15 gene, respectively. The expression of objective gene in vitro was detected by indirect fluorescent antibody technique and immunohistochemistry. The two DNA vaccines were administered by intramuscular leg injection. An animal challenge experiment was carried out to evaluate the immune protective efficacy of the vaccines. The results indicated that DNA vaccines were successfully constructed and the expression of objective gene could be detected in vitro. The animal experimental results showed that both DNA vaccines could provide partial protection against homologous challenge in chickens. The chimeric DNA vaccine, pcDNA3.1-3-1E-linker-mChIL-15, could significantly increase oocyst decrease ratio, reduce the average lesion score in the duodenum, improve body weight gain, and increase anti-coccidial index (ACI) compared to the DNA vaccine pcDNA3.1-3-1E. Taken together, these results demonstrate ChIL-15 enhance the immunogenicity of 3-1E DNA vaccine, and co-expression of cytokine and optimized surface antigen of Eimeria may be a promising method to enhance immunogenicity of DNA vaccines in poultry.     

地方医科大学实验动物管理工作水平评价指标体系的构建

目的构建地方医科大学实验动物管理工作水平评价指标体系,为高校实验动物管理工作的评价提供参考。方法根据研究主题确定咨询专家人选37人,采用Delphi法进行问卷调查和统计分析。结果两轮咨询问卷回收率分别为70．27％和76．92％。权威系数为0．855;第二轮指标体系的协调系数为0．486。结论本次研究构成的地方医科大学实验动物管理工作水平评价指标体系是可靠的。     

铁红圆酵母属两个类群的培养特征初步研究

通过超滤膜过滤法,在大连市黑石礁海域２５００米以内,分离到白色与铁红圆酵母两类海洋酵母。铁红圆酵母分属于Ｔｏｒｕｌｏｐｓｉｓｐｕｌｃｅｒｒｉｍａ类群和Ｔｏｒｕｌｏｐｓｉｓｃａｎｄｉｄａ类群。葡萄糖——蛋白胨——酵母膏液体培养基静止培养条件下,Ｔ．ｐｕｌｃｅｒｒｉｍａ类群培养五天半,活酵母数量达到第一次高峰,为１．２×１０￣８／ｍｌ,大约在八天半之后,达到最高数量,为２．０×１０８／ｍｌ。Ｔ．ｃａｎｄｉｄａ类群,培养大约二天半后,酵母数量达到第一次高峰,为１．８×１０￣７／ｍｌ,培养大约十天半,活细胞数达到最高峰,为１．２×１０￣（－８）／ｍｌ。由于铁红园酵母具有易培性,耐不良环境能力强的优点,作为未来水产养殖业幼体饵料微生物具有广泛地应用前景。     

江西婺源县蜡梅属资源现状及其开发利用

通过踏查法对江西婺源县蜡梅属资源进行调查,阐述了婺源县蜡梅属植物的分布、形态特征、群落学特点和开发利用状况.并提出要加强当地蜡梅的就地保存工作,用人工栽培来取代对野生资源的掠夺,以确保资源的永续利用.     

Mast cell-induced lung injury in mice infected with H5N1 influenza virus

Although an important role for mast cells in several viral infections has been demonstrated, its role in the invasion of highly pathogenic H5N1 influenza virus is unknown. In the present study, we demonstrate that mast cells were activated significantly by H5N1 virus (A/chicken/Henan/1/2004) infection both in vivo and in vitro. Mast cells could possibly intensify the lung injury that results from H5N1 infection by releasing proinflammatory mediators, including histamine, tryptase, and gamma interferon (IFN-γ). Lung lesions and apoptosis induced by H5N1 infection were reduced dramatically by treatment with ketotifen, which is a mast cell degranulation inhibitor. A combination of ketotifen and the neuraminidase inhibitor oseltamivir protected 100% of the mice from death postinfection. In conclusion, our data suggest that mast cells play a crucial role in the early stages of H5N1 influenza virus infection and provide a new approach to combat highly pathogenic influenza virus infection.     

A strategy for fusion expression and preparation of functional glucagon-like peptide-1 (GLP-1) analogue by introducing an enterokinase cleavage site

KGLP-1, a 31-amino acid glucagon-like peptide-1 (GLP-1) analogue, has a great therapeutic potential for anti-diabetes. In this work, a strategy for expression and purification of functional KGLP-1 peptide has been established. KGLP-1 cDNA was fused with glutathione S-transferase (GST), with an enterokinase cleavage site in the fusion junction. The recombinant fusion protein GST–KGLP-1 was affinity purified via the GST-tag, and then digested with enterokinase. The resulting GST part as well as the enzymes were eliminated by ultra-filtration followed by size exclusion chromatograph. The yield of purified KGLP-1 was approximately 12.1 mg/L, with purity of 96.18 %. The recombinant KGLP-1 was shown to have similar bioactivity as native GLP-1 when evaluated in a Chinese hamster ovary cell line expressing a GLP-1 receptor-egfp reporter gene.