旋毛虫谷氨酸脱羧酶基因的原核表达及其活性研究

采用RT-PCR从旋毛虫中扩增出谷氨酸脱羧酶(TsGAD)基因全长,将其克隆到表达载体pET-30a(+)后,转化到大肠杆菌BL21(DE3)中进行IPTG诱导表达,应用比色法鉴定表达产物(rTsGAD)的酶活性,并用纯化的GAD蛋白制备多克隆抗体。结果显示,成功构建了重组质粒pET-30a(+)-TsGAD,经SDS-PAGE分析,rTsGAD蛋白的分子质量约为57ku。利用比色法测得粗提的rTsGAD具有酶活性,酶活力为1.5U。制备的多克隆抗体效价较高,可达1∶65 536。Western-blot分析结果显示,该多克隆抗体与TsGAD具有较强的反应性。结果表明,本试验成功原核表达了rTsGAD,并制备了多克隆抗体,为TsGAD定位以及进一步研究旋毛虫谷氨酸依赖型抗酸机制奠定了基础。     

社区反恐怖视阈下的警务工作探究

《中华人民共和国反恐怖主义法》的正式施行,为我国反恐怖工作的顺利开展提供了法律保障。社区反恐就是以维护社区安全和群众生活稳定、解决社区治安问题和减少犯罪为根本目标。通过分析当前社区反恐存在的主要问题,构建以社区群众为防范恐怖犯罪的主要力量,以基础信息摸排、情报先行、先发制敌为基本原则;以重点人管控、重点部位管理、风险评估等机制建立。同时把社区涉恐情报信息搜集、去宗教极端化、反恐法律宣传、群防群治列为工作重点,积极构建全民反恐的人民战争。     

长三角地区产业向“泛长三角”转移的路径与模式研究

＂十二五＂时期,受成本、资源、环境、政策等因素的影响,长三角地区产业将呈现出大规模向周边乃至中西部地区转移的趋势。考虑各种因素的综合影响,在近期内,长三角地区产业转移的主要区域是＂泛长三角＂地区,借助轴向式转移和圈层式转移两种方式,将会集中将成本驱动型、产业配套型、资源利用型、市场拓展型等四种类型产业沿长江和交通干线向安徽、江西等的泛长三角地区转移。泛长三角地区应积极抓住这一机遇,主动对接长三角,通过承接产业转移实现本地产业的发展和升级。     

China’s ten years in the wto: its performance and new challenges

Becoming a member of the WTO in 2001 was a historic event of great significance during the process of China’s reform and opening up. Since then, China has steadily pushed forward the reform and opening up policy, proactively seizing the opportunities of economic globalization and positively utilizing the multilateral trading system to develop economic and trade relations with other countries, all of which have contributed to the great economic and social achievements during the first decade of 21st century. However, there are different opinions on China’s futuristic role in the WTO, and those disagreements resulting from various interest preferences are not only one-sided and limited, but also triggering off the discussions on the criteria to assess China’s performance in the WTO. This article argues that China’s activities in the WTO (i.e., implementing WTO commitments, participating in the Doha Round negotiation, the dispute settlement and trade policy review) should be a kind of assessment criteria. Based on comprehensive observation of China’s performance in the WTO, it is concluded in this article that China’s participation in the WTO system and global trade governance extends the scope of world trade law, improves its effectiveness, constitutes China’s new contributions to implement treaty obligations in good faith, resolves peacefully international trade disputes, and maintains substantively the international rule of law. At the same time, it has not only caused new driving forces for international trading system, but also made China face new challenges in the WTO.     

猪传染性胃肠炎病毒S-N融合双基因疫苗的构建及其免疫原性分析

为了构建TGEV S-N融合双基因疫苗并分析其免疫原性,从S、N基因克隆质粒中以PCR扩增了S基因(2.1kb,含A、B、C、D抗原位点)和N基因(1.2kb),将S基因插入pVAX1载体构建了pVAX-S质粒,再将N基因插入pVAX-S中S基因末端,构建了融合表达S、N双基因的重组质粒pVAX-S-N,将pVAX-S-N转染COS7细胞以免疫荧光试验进行S、N双基因的表达鉴定。用纯化的pVAX-S-N和作为对照的pVAX-S、pVAX1、PBS免疫BALB/c小鼠,共免疫3次,分别测定免疫后第0、14、28、42天的小鼠血清IgG抗体,测定免疫后第42天小鼠外周血T淋巴细胞亚群(CD3+、CD4+、CD8+)的数量。结果,融合质粒pVAX-S-N可在COS7细胞特异性表达S、N两个蛋白,pVAX-S-N免疫小鼠后第14天即可诱导产生抗TGEV的特异性IgG,但pVAX-S-N诱导的抗体水平一直低于pVAX-S诱导的抗体水平,在免疫后第42天差异极显著(P<0.01);pVAX-S-N可激发小鼠产生细胞免疫应答,但pVAX-S-N组的CD3+、CD4+、CD8+数量均低于pVAX-S免疫组。研究结果表明,融合双基因疫苗pVAX-S-N具有免疫原性,但免疫效果却不如单基因疫苗pVAX-S的理想。     

减毒沙门菌递呈的猪传染性胃肠炎病毒S基因与猪IL-6基因双启动子DNA疫苗的构建

从猪传染性胃肠炎病毒克隆质粒pMD19T-S中扩增了2.1kb的S基因片段(含完整的A、B、C、D 4个抗原位点序列),以RT-PCR从猪外周血淋巴细胞中扩增了猪白细胞介素6(IL-6)(658bp),插入同一双启动子真核表达载体pVAXD的2个多克隆位点,构建了共表达S与IL-6基因的双启动子表达质粒pVAXD-S-IL-6,转染COS7细胞进行转录和表达鉴定。将质粒pVAXD-S-IL-6电转化减毒鼠伤寒沙门菌SL7207,筛选获得重组菌株SL7207(pVAXD-S-IL-6),并对重组菌株SL7207(pVAXD-S-IL-6)的体外稳定性和口服小鼠的安全性进行了鉴定。结果表明该双启动子真核表达质粒pVAXD-S-IL-6构建成功,RT-PCR检测到转染的COS-7细胞中2个目的基因的转录,间接免疫荧光试验结果显示表达产物可与猪抗猪传染性胃肠炎病毒血清发生特异性反应。重组菌株SL7207(pVAXD-S-IL-6)在含卡那霉素的抗性培养基中稳定性好,以1×109 CFU灌胃接种小鼠具有良好的安全性。本研究构建了猪传染性胃肠炎病毒的新型口服疫苗株SL7207(pVAXD-S-IL-6),为开展其免疫原性评价奠定了基础。     

Perceptions and experiences of intrusive behavior and stalking within a culture

ABSTRACT

Previous studies have compared perceptions and experiences of intrusive activity and stalking between countries and the present work compares subcultures within a single country. Singaporean women (89 Chinese, 69 Indian and 68 Malay) with similar age profiles completed a modified version of the ‘Stalking: International perceptions and prevalence’ questionnaire (SIPPQ). This contained measures of individual perceptions and experiences of a range of 47 intrusive behaviors, and a measure of stalking. Between the three ethnic groups, few differences were found in terms of how unacceptable the 47 intrusive acts were judged to be. The small number of differences identified related to courtship behaviors. Women across ethnicities reported largely the same experiences of intrusive behaviors, differing on just two of the 47 behaviors. This suggests that overarching national attitudes towards women are better determinants of the types of intrusive behaviors they are likely to experience, as opposed to their subculture. Finally, no differences were found in stalking rates between the three ethnic groups, but the overall stalking rate within the sample was high at 54.9%. This finding may provide impetus to increase awareness of stalking in Singapore, given that it was only recently criminalized in the sovereign city–state.     

藏区刑事案件发案特点、原因及防控对策--以四川省康定县为调查对象

藏区地理环境特殊,经济、文化、教育发展水平相对落后,其刑事犯罪有自身特点。刑事犯罪的防控工作需要因地制宜。以康定县为例,该县刑事犯罪呈现出较强的季节性特点,案件多与虫草相关,家族仇杀等恶性命案以及赌博、拦路抢劫案件多发,各类劳务、劳资纠葛引发的案件逐渐增多。其刑事犯罪案件发生原因主要有犯罪者个体、职能管理部门乃至社会大环境方面存在的弊端。刑事犯罪的防控需要相应地从上述多方面寻求对策。     

命案现场重建解析—“7·06”灭门惨案现场重建

2004年云南“7·06”灭门惨案得以及时告破,得益于办案人员凭借对案发现场的精勘细验和认真分析研究;是一起典型的重建案发现场,进而一举破案的成功案例。     

论互联网社交在法治社会建设中的功能

作为一个同法治国家、法治政府相区别的概念,法治社会命题致力于实现社会生活的秩序化、法治化,并尤其强调社会力量对国家法治建设的推动作用,试图借助多元社会主体的参与来型构法治发展的动力基础。准确认识当下的社会状况,充分挖掘现有的社会力量,是开展法治社会建设的正确路径。在当代,互联网信息技术的纵深发展日益改变着现代社会的交往模式,并塑造出互联网社交这一新型的社会关系形态。互联网社交的出现与发展孕育了以网络为介质的社会力量,使社会逐渐获取了同国家对话并参与国家法治建设的能力。互联网社交在法治社会建设中的功能可以从四个向度获得理解:互联网社交缔造着网络公共空间的公民品格;互联网社交孕育了网络社会的自治能力;互联网社交构成了民主参与的"非正式"渠道;互联网社交提供了权力制约的社会手段。