猪繁殖与呼吸综合征病毒重组质粒pEGFP-NSP9的构建与表达

为验证siRNAs对猪繁殖与呼吸综合征病毒复制的抑制效果,构建了猪繁殖与呼吸综合征病毒NSP9基因与增强型绿色荧光蛋白基因GFP的融合表达质粒,并在Marc145细胞中进行了表达。通过RT-PCR方法扩增猪繁殖与呼吸综合征病毒NSP9基因,将其克隆入表达载体pEGFP-N1,并进行双酶切、PCR及测序鉴定。将阳性重组质粒转染Marc145细胞,检测绿色荧光蛋白的表达和NSP9基因转录水平。结果显示,经双酶切及PCR鉴定,目的基因的大小与预期相符。荧光显微镜和流式细胞仪均检测到细胞内绿色荧光蛋白的表达,荧光定量PCR检测到细胞内有NSP9基因的转录。本研究成功构建的猪繁殖与呼吸综合征病毒NSP9基因GFP融合表达质粒,为在细胞水平快速筛选有效的siRNAs提供了工具。     

猪繁殖与呼吸综合征疫苗的研究进展

简要介绍了国内外猪繁殖与呼吸综合征弱毒疫苗、灭活疫苗和基因工程疫苗的研究进展,分析了猪繁殖与呼吸综合征疫苗研制中存在的问题和未来发展趋势,以便兽医药科技工作者全面了解猪繁殖与呼吸综合征疫苗的研发及使用现状,进而指导临床应用.     

猪繁殖与呼吸综合征病毒引起断乳仔猪水肿病

猪繁殖与呼吸综合征（ＰＲＲＳ）是８０年代新出现的一种传染病。临床主要表现为母猪的繁殖障碍及仔猪与育成猪的呼吸系统症状。但由于猪的年龄、免疫状态以及毒株的不同影响着发病机理,使猪繁殖与呼吸综合征病毒（ＰＲＲＳＶ）感染后表现出多种不同的临床症状。我们从某...     

河北省猪繁殖与呼吸综合征病毒隐性感染情况的调查

为了解河北省猪繁殖与呼吸综合征病毒(PRRSV)的隐性感染情况,采用RT-PCR方法对2006-2009年采自全省11个地市228个县级屠宰场1 565份商品猪的肺或肺门淋巴结样品进行了PRRSV病原学检测。结果显示,PRRSV场总体阳性率为45.18%;高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)样品总阳性检出率为12.08%,这4年样品的阳性检出率分别为28.45%、40.74%、12.01%和7.31%;经典猪繁殖与呼吸综合征病毒(C-PRRSV)样品总阳性检出率为2.36%,这4年样品的阳性检出率分别为3.45%、3.70%、3.92%和1.46%。PRRSV同猪瘟病毒、猪伪狂犬病病毒和猪圆环病毒2型混合感染所占的比率为59.73%,HP-PRRSV和C-PRRSV同其他病原体混合感染所占的比率分别为57.67%和43.24%,混合感染以二重或三重感染为主。在同一份样品中不能同时检测到HP-PRRSV和C-PRRSV。总之,PRRSV,尤其是HP-PRRSV在河北省猪群中隐性感染的现象普遍存在,值得高度重视。     

猪繁殖与呼吸综合征病毒的分离及其基因组变异特征的研究

为了解猪繁殖与呼吸综合征病毒(PRRSV)在河南省新乡地区猪体内的进化情况,笔者对河南省新乡地区某猪场送检的疑似猪繁殖与呼吸综合征的病料进行了病毒的分离鉴定,并采用PCR扩增得到了该病毒的全基因组序列,同时对分离病毒株的GP5和NSP2序列进行了比对分析。结果显示:成功分离并鉴定出1株美洲型PRRSV(XX2015),与中国代表毒株CH-1a中的核苷酸同源性是92.8%,与高致病性(HP)-PRRS病毒代表株Hu N4、Hu B1、He Nan-1、He NAN-A14株的核苷酸同源性则分别为97.2%、97.0%、97.2%、94.4%。对基因组的进一步分析表明,该毒株GP5蛋白非中和性表位处存在氨基酸变异,NSP2蛋白在第481位、第485~488位、第533~561位共缺失34个氨基酸。XX2015分离株是HP-PRRSV的一个代表株,该病毒株在NSP2蛋白中存在跳跃性突变,为进一步研究猪繁殖与呼吸综合征病毒的分子进化提供了分子证据。     

猪繁殖与呼吸综合征病毒样颗粒的构建

为构建高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)病毒样颗粒(VLPs)疫苗,以分离鉴定的HP-PRRSV CQ株的GP5和M蛋白的全部编码基因为模板,利用杆状病毒-昆虫细胞表达系统,共表达GP5蛋白和M蛋白,并采用SDS-PAGE、Western-blot对重组蛋白进行定性分析;通过透射电子显微镜对病毒样颗粒进行直观的形态学鉴定。结果表明,利用杆状病毒-昆虫细胞表达系统能够成功地共表达重组质粒pFastBac1-GP5-M的GP5蛋白和M蛋白,并能自动装配成病毒样颗粒,显示具有与天然蛋白类似的免疫原性;电镜观察到病毒样颗粒的形态与文献中描述一致,直径约为50nm。本试验结果为猪繁殖与呼吸综合征病毒样颗粒疫苗的研究奠定了基础。     

猪圆环病毒病及其生物制品学的研究进展

结合国内外对猪圆环病毒病的研究结果,概述了猪圆环病毒病的主要临床类型及其流行病学特点,并对其病原学、免疫学及生物制品学方面的研究进展进行了综合论述,为认识该病并有效防控该病提供参考。认为,猪圆环病毒2型的危害主要体现在断奶仔猪多系统衰竭综合征症状中,并与猪细小病毒、猪繁殖与呼吸综合征病毒、副猪嗜血杆菌、猪肺炎支原体等的继发感染有关。     

猪繁殖与呼吸综合征病毒GP5蛋白糖基化位点缺失突变株的构建

以含有猪繁殖与呼吸综合征病毒(PRRSV)VR-2332株全长感染性cDNA克隆的质粒pVR2332为模板,通过融合PCR方法扩增GP5蛋白第34、51位天冬酰胺糖基化位点缺失的突变序列,替换pVR2332质粒中编码GP5蛋白的DNA片段,分别获得重组质粒pVR-N34A、pVR-N51A。在体外转录后转染BHK-21细胞,转染后第48小时收集细胞接种Marc-145细胞,进行突变病毒的拯救。然后通过RT-PCR扩增拯救病毒GP5蛋白的编码序列,序列分析显示构建正确。间接免疫荧光试验证明拯救病毒能够正常表达GP5蛋白。同时,中和试验结果初步表明GP5糖基化位点缺失影响PRRSV与中和抗体的反应。猪繁殖与呼吸综合征病毒GP5蛋白不同糖基化位点缺失突变株的获得将为进一步揭示GP5蛋白糖基化与诱发机体免疫反应之间的关系奠定基础。     

同时检测五种引起猪繁殖障碍的病毒的多重PCR的建立

为建立一种能同时检测5种引起猪繁殖障碍的病毒的多重PCR方法,分别针对各病毒的保守序列设计了4对特异性引物,其中3对分别用于扩增猪瘟病毒(CSFV)E2基因、非洲猪瘟病毒(ASFV)VP72基因、伪狂犬病病毒(PRV)gB基因的目的片段;针对NSP2基因设计的1对引物用于区分猪繁殖与呼吸综合征病毒(PRRSV)和高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)。通过对反应条件的优化,建立了快速检测CSFV、ASFV、PRV、PRRSV和HP-PRRSV的多重PCR方法。敏感性试验结果显示,该多重PCR对这5种病原核酸的最低检测量分别为2.10×103(HP-PRRSV),1.30×103(PRRSV),1.09×104(CSFV),1.50×103(ASFV)和8.97×102(PRV)copies/μL。对49份临床样品的检测结果显示,它们的混合感染率为51%(25/49)。该方法对阴性样本的扩增结果均为阴性,并且无交叉反应性,表明该方法特异性良好,具有快速、灵敏且成本低等优点,能够对猪繁殖障碍病毒病的单个或混合感染的临床样品进行快速鉴别诊断,并对其流行病学调查具有深远意义。     

南阳市猪繁殖与呼吸综合征流行情况调查

南阳市猪繁殖与呼吸综合征流行情况调查石应俭张国宪牛鑫王灵敏刘志华王国俊常万有（河南省南阳市畜牧兽医工作站４７３０００）猪繁殖与呼吸综合征（ＰＲＲＳ）是一种导致妊娠母猪流产、仔猪和育肥猪发生肺炎为特征的病毒性传染病。目前，该病已遍布世界各地的养猪地区和...     

Undermining cooperation: donor-patrons and the failure of political conditionality

This study extends the debate concerning the feasibility of using foreign assistance to leverage political reforms in developing countries. This research combines both quantitative and qualitative methodologies in an effort to expand on the work done by previous scholars. Conclusions of the empirical analysis point to the finding that political conditionality is most effective when close cooperation exists between donors. This coordination, however, is frequently sabotaged by self-interested donor-patrons, illustrated by the case of Cameroon and France. This pattern is generalizable and significantly undermines the overall success rate of a political conditionality approach.     

European small states' military policies after the Cold War: from territorial to niche strategies

This article looks at the evolution of European small states' military policies after the Cold War. Traditionally, small states faced a security dilemma between favouring influence and guaranteeing sovereignty. These security options were embodied by the strategy of alliance and the policy of neutrality. This article argues that in today's unipolar world small states' security policy must be cooperative either in the form of joining a security institution or an ad hoc coalition. This has two consequences for small states' military policies. These can either favour niche or lead/framework nation strategies. This in turn, depends on the strategic ambitions of the small states, which are ultimately mediated by their strategic culture. This article concludes by looking at the military policies of Cold War neutral states after the Cold War.     

Shaping participation: an international NGO implementing a government participation policy

This article reports on a case study of how one NGO utilised a government participation policy to establish ostensibly participatory spaces in the Cambodian health sector. The ethnographic field study revealed how the NGO exerted influence in establishing and facilitating participative committees by controlling membership, resources, and meeting agendas. This resulted in limited citizen participation, with committees used to educate, lobby, and mobilise community leaders to work towards the NGO’s priorities rather than community identified needs. This case contributes to our understanding of insisted spaces and the role of third parties implementing government participation policies.     

Countering Violent Extremism: A Realist Review for Assessing What Works,for Whom,in What Circumstances,and How?

ABSTRACT

Scientific knowledge on what works in countering violent extremism remains limited. This article argues that we should move away from the “what works?” question and towards: “what works, for whom, in what circumstances, and how?” This method is also known as realist evaluation. This article applies the realist review method to CVE studies, which synthesizes the existing CVE literature and helps us gain insight into relevant contexts, mechanisms, and outcomes for CVE. Realist reviews help to develop and shape more effective policy and contribute to further CVE theory development.     

Good intentions are not enough: French NGO efforts at democracy building in Cameroon

NGOs have traditionally had little scope to bring about political reform in developing countries. This was certainly true of French development NGOs (NGDOs) operating in Cameroon during the early post-colonial decades. This situation changed in 2002 when French NGDOs, with support from the French state and Cameroonian civil society, initiated a multi-actor consultative programme (the PCPA), aiming to build democracy in Cameroon. This article traces the origins of the PCPA, assesses its achievements, and explains why the programme failed. It then identifies key lessons and asks whether the PCPA represents a useful model for French NGOs and donor states.     

Who Are Pakistan's Militants and Their Families?

This article presents results of a survey of 141 Pakistani families of slain militants. This survey collected data about the militants and their households. While derived from a convenience sample, these data are unprecedented and offer a glimpse into the backgrounds of militants and the families who (mostly) supported their decision to join the jihad. Most militants served and died in Kashmir and seem to be “high quality” militants in that they, like their heads of household, are well educated and not predominantly coming from seminaries, as is often claimed. This analysis suggests that while the militants merit attention, so do the families that produce militants.     

THE RELATIONSHIP BETWEEN NEW SOCIAL MOVEMENT THEORY AND TERRORISM STUDIES: THE ROLE OF LEADERSHIP,MEMBERSHIP, IDEOLOGY AND GENDER

This article demonstrates the relationship between new social movement theory and terrorism studies. The revolutionary dimension of new social movements can be correlated to terrorist groups. This article also uses new social movement theory to develop a fuller understanding of the leadership, membership, personal ideology and gender within a new social movement; these then can be used to follow the development of a politically violent group. This article uses the example of the American movement and its by-product—the Weather Underground.     

The remote securitisation of Islam in the US post-9/11: euphemisation,metaphors and the “logic of expected consequences” in counter-radicalisation discourse

This article critically analyses the securitisation of Islam post-9/11 in the US and argues that this securitisation is a remote securitisation whereby the securitisers – the security practitioners – are placed at a distance from the securitisees – the Muslim community. This is achieved through two processes of security practice: linguistically by euphemising language and using metaphors, and analytically by understanding radicalisation through a rationalist perspective, which follows the “logic of expected consequences”. This article further problematises the rationalist view of radicalisation in the counterterrorism sector in the US and concludes by introducing a Bourdieusan concept of relationality to critical counter-radicalisation studies.     

东盟中国“10+1”关系的全方位发展——纪念中国—东盟建立对话伙伴关系20周年

今年是中国和东盟建立对话伙伴关系20周年.20年来,双方关系得到了深入和广泛的发展,东盟中国"10+1"关系更是在全方位地发展.本文阐述东盟中国"10+1"关系的发展,分析其特点和作用,并展望双方关系未来的发展.     

马来西亚环保非政府组织浅析

摘要：马来西亚环保非政府组织是马来西亚众多非政府组织中发育良好并卓有成效的公民社会组织,但学术界对马来西亚环保非政府组织的研究散见于各种文献之中。本文尝试从多个方面对马来西亚环保非政府组织进行较为综合的介绍和分析。首先,本文介绍了马来西亚环保非政府组织的发展概况,认为他们不仅是马来西亚环保运动的先驱者,而且也是发展中国家环保运动的先驱者之一;接着,介绍了主要环保非政府组织的基本情况;然后,以环保非政府组织反对和监督巴昆大坝建设为主要案例,对马来西亚环保非政府组织的主要活动及环保非政府组织与政府的关系进行了介绍和分析。