Expression of a Shrimp Kunitz-type Protease Inhibitor in Pichia pastoris and Activity Analysis

SKPI(shrimp Kunitz-type protease inhibitor)是日本囊对虾(Marsupenaeus japonicus)体内的一个小分子多肽,含有一个Kunitz型结构域,属于丝氨酸蛋白酶抑制剂。目前已知丝氨酸蛋白酶抑制剂在节肢动物免疫系统中起着非常重要的作用,为了了解SKPI在对虾天然免疫系统中的作用,首先对其进行了重组表达。从日本囊对虾肝胰腺中扩增skpi的cDNA片段,插入改造后的pPIC9K酵母表达载体,获得的重组质粒转化至毕赤酵母GS115进行表达。由于改造的pPIC9K载体加入了6-His标签,因此利用Ni Sepharose High Performance对SKPI进行了高效纯化。初步的活性研究表明,重组表达的SKPI能特异性地抑制胰蛋白酶的水解活性。SKPI(shrimp Kunitz-type protease inhibitor) from Marsupenaeus japonicus is a member of serine protease inhibitors which play an important role in the arthropod immunity.To fully understand its function in the innate immunity of shrimp,the skpi gene was cloned into a modified pPIC9K vector with a 6-His tag and expressed by Pichia pastoris GS115.The secretory SKPI was purified from the medium with high purity by using Ni Sepharose High Performance.This results also indicated that the purified SKPI could inhibit the activity of trypsin specifically.国家高技术研究与发展计划项目(No.2006AA100311);; 福建处科技厅资助重点项目(No.2006N0039)~

俄罗斯的太空安全观

太空被视为俄罗斯的国家总体实力与军事实力的象征。维护太空安全与太空力量的平衡,成为俄罗斯国家战略考虑的问题。面对美国太空武器化的政策,为保证其进出太空的自由和其太空战略利益,俄罗斯对此做出回应。一方面,联手中国等国家呼吁国际社会限制太空武器化;另一方面谋求"非对称反击"手段,发展相关反制武器,不让美国在太空武器领域一枝独秀

Application and Development of Synchronous Fluorescence Spectrometry

［中文文摘］同步荧光技术是解决多组分荧光物质同时测定的良好手段之一。本文从恒波长同步荧光法、恒能量同步荧光法、可变角同步荧光法、恒基体同步荧光法以及它们与导数技术、低温技术、化学计量学方法的联用等方面对同步荧光技术领域出现的新技术及应用作一评述。［英文文摘］Synchronous fluorescence spectrometry is useful in simultaneous determination of multi-component fluorescent substances. Its application and development in recent years are reviewed. Synchronous fluorescence approaches, including constant-wavelength synchronous fluorescence, constant-energy synchronous fluorescence, variable-angle synchronous fluorescence, isopotential matrix synchronous fluorescence, and the combination with the derivative technique, stoichiometry and cryogenic technique are discussed.国家自然科学基金(No .2 9875 0 2 3);教育部优秀青年教师基金;福建省自然科学基金资助项目

几种天然产物在海洋防污涂料中的应用研究

将天然产物齐墩果酸、蛇床子素和鱼藤酮作为防污剂,分别制备海洋防污涂料,在厦门海域检验其海区防污效能,并与已报道在厦门海域具防污效能的天然产物喜树碱进行比较,发现蛇床子素和鱼藤酮在海区挂板2个月内显示出一定的防污效能,齐墩果酸没有呈现出防污效能,而喜树碱的海区防污效能明显优于上述3种天然产物。另外,将喜树碱与鱼藤酮进行1∶1复配,并将这2种天然产物与氧化亚铜、N-(2,4,6-三氯苯基)马来酰亚胺(TCPM)和吡啶硫铜锌(ZPT)这3种防污(助)剂分别进行1∶1复配,共获得7种复配防污剂,制备相应的海洋防污涂料以检测其防污效能,发现喜树碱-TCPM、喜树碱-ZPT以及喜树碱-鱼藤酮这3种复配防污剂具良好的防污效能。进一步在上述涂料中选取了防污效能最优的含喜树碱涂料,在福建东山海域和海南陵水海域分别进行了海区挂板试验,结果表明含喜树碱涂料在这2个海域均显示出稳定、优异的防污效能,且期效可达13个月以上。文中研究为推进天然防污产物在海洋防污涂料中的应用进程提供了重要资料。厦门市海洋经济创新发展示范产业链协同创新类项目(16CZB023SF12);;海洋(赤潮)灾害立体监测技术与应用国家海洋局重点实验室开放研究基金(MATHAB201808);;福建省海洋经济发展补助资金项目(FJHJF-L-2018-2

Effects of Diosmetin from Carbonized Cirsium japonicum on Apoptosis in Human MCF-7 Breast Cancer Cells and its Mechanisms

研究大蓟炭中香叶木素诱导人乳腺癌MCF-7细胞凋亡及其作用机制。采用硅胶和Sephadex; LH-20柱层析从大蓟炭中分离鉴定了三个黄酮类化合物,经NMR和MS鉴定他们的结构分别为香叶木素(1)、刺槐素(2)和柳川鱼黄素(3)。采用MT; S方法检测不同浓度的香叶木素对MCF-7的细胞活力的影响;流式细胞术检测不同浓度的香叶木素处理对MCF-7细胞凋亡的作用;Western; blot法检测香叶木素处理对细胞PARP、P-JNK等细胞凋亡相关蛋白表达的影响。MTS及流式细胞术结果显示香叶木素能显著抑制MCF-7的增殖并; 且诱导细胞凋亡;香叶木素可上调P-JNK促进细胞凋亡。结果表明香叶木素在体外实验能通过激活JNK细胞凋亡通路抑制MCF-7的增殖及促进细胞凋亡。To study the effects of diosmetin from carbonized Cirsium japonicum on; apoptosis in human MCF-7 breast cancer cells and its mechanisms. Three; flavonoids were isolated and purified from carbonized C. japonicum by; silica gel and Sephadex LH-20 chromatography methods. Their structures; were elucidated by NMR and MS spectroscopic data and identified as; diosmetin(1),acacetin(2) and pectolinarigenin(3). MTS assay was; performed to detect the viability of MCF-7 cells treated by different; concentrations of diosmetin. The cell apoptosis rate was further; analyzed by flow cytometry(FCM). Western blot assay was applied to; measure the apoptosis related protein expression levels of PARP,P-JNK.; Diosmetin treatment on MCF-7 cells significantly inhibited cell; proliferation and induced cell apoptosis. Diosmetin significantly; downregulated P-JNK and upregulated cleaved-PARP protein expression.; Diosmetin inhibited MCF-7 cell proliferation and induced cell apoptosis; by activation of JNK pathway in vitro.江西中医药大学校级科研项目; 国家自然科学基金; 江西省自然科学基

改良分子信标-双重实时荧光PCR快速检测SARS病毒

目的建立改良分子信标-双重实时荧光PCR检测SARS病毒的方法,用于SARS的早期诊断和动物溯源。方法利用改良分子信标技术、装甲RNA和双片段双色荧光技术,根据GenBank公布的SARS病毒聚合酶基因1b的阅读开放框架结构的保守序列,自行设计一对引物和探针,以部分临床标本的酶联吸附实验结果和传统细胞培养方法作为对照,建立分子信标检测SARS病毒的方法。对368份临床标本(咽漱液、血液、粪便、尿液)、52份细胞培养液和50份动物标本进行荧光PCR扩增。结果分子信标检测SARS病毒的方法灵敏度为10～100个拷贝ml,与流感病毒等呼吸道病毒无交叉反应。分子信标检测368份临床标本,20份阳性。其中确诊病例阳性率为21.27%(1047),确诊病例的咽漱液阳性率为43.48%,还分别从粪便和血清中检测到SARS病毒。52份细胞培养液,29份阳性,阳性率为55.77%。50份动物标本,23份阳性,阳性率为46%。结论改良分子信标-双重实时荧光PCR检测SARS病毒方法灵敏度高、特异性强,可用于SARS的临床早期诊断和动物溯源

Clinical and pathological significance of HPV infection and p53 mutation in human esophageal cancer

published_or_final_versionAnatomyDoctoralDoctor of Philosoph

Discussing on the Cultivation Scheme for Compound-ability Professionals in International Relations

如何培养具有中国特色、精通理论、熟稔政策具有国际眼光与专业水准的国际关系专业复合型人才,这是目前国内高校与研究机构的国际关系专业面临的时代要求。随着中国国际地位的提升,纯粹西方学术话语体系下的西方国际关系理论,特别是经典的三大主义研究范式,已经无法满足中国外交与国际关系研究与实践的客观需求。以国际关系理论中国学派生成为出发点,在教学实践中,探索以问题意识为导向,跨学科研究为支撑的复合型专业人才培养模式。How to cultivate the compound-ability professionals,who master theory,are familiar with policy and characterized with Chinese feature,is the theme of current era faced by domestic universities and institutions.With the rise of China's international status,the international relation theory purely rooted in the western academic discourse system,especial realism idealism and constructivism paradigm,does not meet the requirement of the theory and practice of China's diplomacy and international relations any more.From the perspective of building Chinese school,this paper explores the cultivation mode for compound-ability professionals in international relations,oriented with problem-consciousness and supported with cross-disciplinary research.上海政法学院国际政治专业教育高地建设项目资

对两种期刊近十年技术史论文的统计与分析

用统计学的方法对《中国科技史料》和《自然科学史研究》两种重要科学技术史期刊1994~2005年发表的中国技术史论文进行统计与分析,对近十年来中国技术史研究的主要领域及其进展进行综述,并对中国技术史研究方法的变化进行总结,从而就中国技术史的发展提出几点建议