MDCK细胞感染犬细小病毒后的差异表达谱分析

通过了解MDCK细胞感染犬细小病毒(CPV)后基因表达水平的差异,研究病毒对细胞的致病作用以及细胞抵御病毒感染的机制。利用表达谱基因芯片技术,分析持续感染犬细小病毒的MDCK细胞基因表达水平的变化情况,并用Real-Time PCR技术加以验证。结果显示,获得了359个差异大于1.5倍的表达基因(P0.05),占总基因数的1.53%,其中193个上调表达(0.84%),166个下调表达(0.69%)。对差异基因进行GO功能聚类分析,小部分涉及免疫应答、生长周期调控、信号转导和蛋白酶活性,其他大部分基因功能未知。利用Real-Time PCR随机验证5个基因在持续感染CPV后的差异表达,其结果和芯片杂交的结果一致。表明建立了MDCK细胞持续感染CPV后的差异表达谱,初步了解到CPV对宿主细胞的制约作用,引起部分细胞增殖调控相关基因发生了表达下调,以及细胞对感染的积极应答反应,部分免疫反应基因、肽链内切酶活性基因等发生了上调表达,从而为探索病毒的致病机理和宿主的抗病毒途径提供了试验基础。     

鸭肝炎病毒VP1和VP3基因的克隆及其杆状病毒表达载体的构建

为了构建鸭肝炎病毒VP1和VP3结构蛋白基因的杆状病毒表达载体,以RT-PCR方法扩增鸭肝炎病毒的VP1和VP3基因,克隆至pGEM-T载体。经筛选测序验证后,以BamHⅠ+XhoⅠ双酶切回收VP1和VP3目的片段,与经相同方法处理的杆状病毒转座载体pFastBac1连接,得到重组质粒pFastBac1-VP1和pFastBac1-VP3。将经酶切及测序鉴定的阳性重组质粒转化入含穿梭载体Bacmid的感受态细胞DH10Bac,发生转座作用,经抗性及蓝白斑筛选,成功获得各自携带VP1和VP3基因的重组穿梭载体,将其命名为rBacmid-VP1、rBacmid-VP3。研究结果为进一步在昆虫细胞中表达VP1或VP3基因,开发研制鸭肝炎病毒基因工程疫苗奠定了基础。     

中华鳖TLR2部分序列的克隆及其组织表达差异分析

为探究中华鳖天然免疫的分子机制,针对TLR2(Toll-like receptor 2)保守区设计兼并引物,从中华鳖脾cDNA中扩增出了740bp的目标序列。测序和序列分析结果表明,该序列包含富含半胱氨酸型C端结构域、跨膜结构域和TIR结构域;与斑马鱼、鸡等脊椎动物相应的TLR2序列相似性为60.5%～74.4%。采用荧光定量PCR检测中华鳖感染嗜水气单胞菌后各组织TLR2mRNA表达的变化,结果显示,感染后第24小时,中华鳖脾和外周血TLR2mRNA相对表达水平升高,其中外周血中的增幅最显著,是对照组的8.57倍;中华鳖外周血经4μg/mL酵母聚糖刺激后,第2小时TLR2mRNA相对表达水平较高。首次获得了中华鳖TLR2部分cDNA序列,并分析了不同刺激物对中华鳖TLR2mRNA转录的影响。     

乡村治理变迁与新乡贤的当代表达

随着我国乡村社会由现代的逐步转型,乡村社会形态在变迁过程中呈现出农业社会、工业社会、现代社会相互叠加的异像,乡村的经济、社会、秩序、道德都进入破碎-重构的通道面临重重困境,作为国家治理重要环节的乡村治理面临重重困境。乡贤文化依附强烈的地缘、血缘、亲缘关系而生而长,内涵着特定场域内的风俗、人情、道德、礼法、情感、习惯及信仰,是我国千百年来乡村治理经验与智慧的宝贵结晶。当代新乡贤因其在治理传统、身份特性、能力资源、行为标尺、品德垂范等方面的“有用性”和“可用性”,为解构乡村治理困境提供了新的思路。     

成文宪法中的宗教内容比较研究

宗教信仰自由权是世界范围内各成文宪法保护的重要内容。与其他人权的宪法保护不同,宗教信仰自由权在各国宪法中具有扩散性保护特征。只有综合考量宪法规范中的宗教内容,才能准确理解宗教信仰自由权的宪法规范保护。通过在各成文宪法中查找与宗教相关的关键词,可以发现宗教信仰自由权成文宪法保护具有普遍性。以宗教信仰自由权为核心,辅之以政教关系的相关规定,构成了宗教问题宪法保护的基本框架。从内容上看,成文宪法保护的宗教信仰自由权具有内容的绝对性与相对性相统一、主体自主选择与结社自由相结合的特点;从属性上看,宗教信仰自由权具有入世性与政治性并存的特征。     

Editorial Introduction

Abstract

There has always been a tension between a critique of ‘real existing conditions’ and meta-theoretical paradigms through which the tasks of critique can both be anchored and images of humankind explored.     

“Willing the Event”

Abstract

A major problem threatens Deleuze’s project in The Logic of Sense. He makes an ontological distinction between events and substances, but he then collapses a crucial distinction between two kinds of events, namely, actions and mere occurrences. Indeed, whereas actions are commonly differentiated from mere occurrences with reference to their causal dependence on the intentions of their agents, Deleuze asserts a strict ontological distinction between the realm of causes (including psychological causes) and the realm of events, and holds that events of all types are incorporeal happenings which are inseparable from expressed sense. For Deleuze, what counts as one’s action thus does not depend on one’s intention, but rather on a process of “making sense” of that action. Nevertheless, Deleuze continues to speak of the need to “will” the event. In order to resolve this apparent contradiction, I will read a conception of “expressive agency” into The Logic of Sense.     

Regulating the Media: Some Neglected Freedom of Expression Issues in the United Kingdom's Counter-Terrorism Strategy

ABSTRACT

In the wake of the first ever Al Qaeda-inspired bombings in Britain in July 2005, there has been much discussion about the appropriate form(s) of counter-terrorism response. This article focuses on one aspect of the “war on terror” usually afforded less prominence than other counter-terrorist measures; namely a range of existing and proposed constraints on media freedom and the constitutional/human rights issues provoked. The United Kingdom is the focus because terrorism laws intended for the ethno-nationalist conflict in Northern Ireland have been replaced by legislation in 2000 and 2001 claimed to reflect the changed nature of terrorism and that arguably has serious implications for freedom of expression. Measures that would impact adversely on speech are being debated in Parliament presently, measures that go considerably further than the previous bans on the direct broadcasting of Sinn Féin representatives and their sympathisers.     

柔嫩艾美球虫pEtK2抗原基因的克隆表达及其表达产物的抗原性分析

应用RT-PCR技术从纯化的柔嫩艾美球虫孢子化卵囊子孢子中克隆了pEtK2抗原基因,并进行了原核表达和抗原性分析,同时用MTT法检测了重组蛋白对球虫耐过鸡淋巴细胞的刺激效果。结果显示,克隆的pEtK2基因全长1464 bp,具有一个完整的开放阅读框,编码487个氨基酸,具有多个T细胞表位。重组蛋白诱导5 h后表达量最多,分子质量约55 ku,占菌体总蛋白的32%。纯化的重组蛋白能够刺激球虫耐过鸡T淋巴细胞增殖,可作为球虫基因工程疫苗或DNA疫苗的候选基因。     

法眼看辽宁西丰“拘传记者案”

辽宁西丰"拘传记者案"是地方公共权力侵犯公民宪法权利,侵害传媒新闻自由权,侵犯记者人身自由权的典型案例。该案例对我们的启示是,应着重在制度层面上调整地方党政机关职权划分和完善党对地方事务,尤其是司法事务的领导方式;在法律层面上进一步完善《刑法》、《刑事诉讼法》等相关法律,细化公安机关和检察机关司法行为的法定要件,特别是要尽快完善关于记者权利保护的立法等,以更好地保障新闻媒体和社会大众对我国各级国家机关及其工作人员进行舆论监督。