残缘璃眼蜱subolesin基因的原核表达及表达产物的反应原性分析

为研究残缘璃眼蜱subolesin基因的生物学特性以及寻找新的抗蜱和蜱传病疫苗候选抗原,根据GenBank上登录的小亚璃眼蜱subolesin基因核苷酸序列设计特异性引物,以残缘璃眼蜱、亚洲璃眼蜱、微小牛蜱的饥饿成蜱cDNA为模板,扩增出subolesin基因的完整阅读框(ORF)。将残缘璃眼蜱subolesin基因的PCR产物经EcoRⅠ+NotⅠ双酶切后连接至表达载体pGEX-4T-1,把重组质粒pGEX-4T-1-subolesin导入感受态细胞BL21(DE3)中,在37℃、1mmol/L IPTG条件下诱导表达。利用SDS-PAGE分析subolesin基因的体外表达特征;用Western-blot分析Subolesin蛋白的反应原性。结果显示,残缘璃眼蜱、亚洲璃眼蜱、微小牛蜱subolesin基因的ORF长度分别为492、492、486bp,分别编码163、163、161个氨基酸。subolesin基因序列和推导的氨基酸序列的比对结果显示,残缘璃眼蜱、亚洲璃眼蜱、微小牛蜱与参考的小亚璃眼蜱的核苷酸序列同源性分别为98%、98%、89%,氨基酸序列同源性分别为98%、99%、93%。SDS-PAGE和Western-blot结果显示,重组残缘璃眼蜱Subolesin蛋白的分子质量为45ku,纯化的subolesin重组抗原可与抗残缘璃眼蜱全蜱血清、残缘璃眼蜱唾液腺、卵巢血清有较强的反应原性。     

尤氏泰勒虫OPRT基因的优化表达

为克隆和表达尤氏泰勒虫OPRT基因的功能区片段,并对重组OPRT蛋白的反应原性进行分析,以反转录得到的尤氏泰勒虫cDNA链为模板,PCR扩增出OPRT基因片段,然后将其克隆至pET-30a(+)载体;把构建成功的重组质粒转化至感受态大肠杆菌BL21进行优化表达,对诱导表达的蛋白纯化后用Western-blot分析它与吕氏泰勒虫和绵羊泰勒虫阳性血清是否发生交叉反应。结果显示,OPRT基因获得了不可溶性表达,最佳表达时间为6h,最佳表达温度为28℃,且表达的重组OPRT蛋白分别与上述2种泰勒虫阳性血清发生交叉反应。结果表明,OPRT同源基因也存在于吕氏泰勒虫和绵羊泰勒虫之中。     

行政诉讼司法建议制度的功能衍化

行政诉讼司法建议原本是一项裁判执行措施,但随着最高人民法院司法政策的变化,其发挥的事实功能被大大扩展。实践中,行政诉讼司法建议在规范性文件修改中发挥着功能性审查的作用。从社会变迁的视角观察不难发现,维稳压力、协调和解政策与地方发展型政府的模式变迁,诱发了对于行政诉讼司法建议的制度性需求,从而迫使司法建议成为行政诉讼工具箱中的重要工具。行政诉讼司法建议制度的功能衍化,为法社会学研究提供了一个极具价值的制度样本,亦为行政诉讼法的未来发展提供了背景材料。     

205例高坠案件局部机械性损伤与死亡性质关系分析

目的探讨高坠案件中局部机械性损伤行为心理分析与死亡性质的关系,为此类案件的定性提供参考。方法收集2008—2013年高坠死亡案件311例,其中存在高坠难以形成的局部机械性损伤案例205例。对损伤形成特点、现场痕迹、高坠致命伤、文字信息等进行分析。结果 205例中,根据初步现场痕迹、高坠致命伤、文字信息等分析后,明确自杀86例,意外24例,性质不明95例。95例性质不明案例经对局部机械性损伤进行行为心理分析后,认定自杀80例,意外11例,他杀4例。结论高坠难以形成的局部机械性损伤与死亡性质具有关联性,根据局部机械性损伤行为心理分析推断高坠死亡性质基本准确。     

A Simple Digestion Method with a Lefort Aqua Regia Solution for Diatom Extraction

Presence of diatoms in tissues has been considered as a significant sign of drowning. However, there are limitations in the present extraction methods. We developed a new digestion method using the Lefort aqua regia solution (3:1 nitric acid to hydrochloric acid) for diatom extraction and evaluated the digestive capability, diatom destruction, and diatoms' recovery of this new method. The kidney tissues from rabbit mixed with water rich in diatoms were treated by the Lefort aqua regia digestion method (n = 10) and the conventional acid digestion method (n = 10). The results showed that the digestive capability of Lefort aqua regia digestion method was superior to conventional acid digestion method (p < 0.01); the structure of diatom remained almost intact; and the recovery of diatom was comparable to the conventional acid digestion method (p > 0.05). The Lefort aqua regia reagent is an improvement over the conventional acid digestion for recovery of diatoms from tissue samples.     

快速PCR扩增检验体系的建立及技术指标验证

目的建立PCR快速扩增程序和体系,并对其技术指标进行验证。方法运用快速扩增酶Fast Start与DNA TyperTM15 plus primer mix组合并优化建立快速扩增体系,对50份静脉血卡和35份案例检材提取的DNA进行扩增,并对体系检测结果的准确性、稳定性和检材适应性进行验证;采用不同稀释浓度的标准品9947,采用快速扩增方法进行检测,验证体系的灵敏度。结果模板DNA浓度达到0.50ng/μL,采用快速扩增体系可能获得准确分型;扩增耗时仅为69min,比常规扩增方法明显缩短;不同种类检材,经检测均获得良好分型图谱,同一份样本重复实验结果一致、稳定。结论本文建立的快速扩增体系可显著提升检测速率,其灵敏度、准确性、稳定性、检材适用性等技术性能,可满足实际检验的需求。     

Introduction: The DPP in power

The transition of power from the KMT to the DPP in Taiwan has not been easy. Instead of bringing in a new way of thinking, it is noted that ideology is too ingrained for the new ruling party to make a break from it. Instead of providing a new form of governance, the DPP has proved to be inexperienced at coping with the intricacies of a modern democratic polity. Rather than building new state-civil society relations, the DPP has been unable to expand its outreach to segments beyond its traditional constituencies. Most painful has been the effort required to try to change the general mindset from confrontational to accommodative. The DPP went on to form a minority government and to challenge the still powerful now-opposition KMT in almost every important policy debate. Consequently, components for effective governance such as coordination, compromise, institution-building, etc., are rare stock. The inherent factional schism is hurting the party as virtues like coherence of policy, coordination of bureaucracies, and even channel of command are compromised. Ethnic incongruence seems to be aggravating the situation.     

关于贪污贿赂案件中证据认定和使用问题的实践分析和理论反思

贪污贿案件中的证据具有言词多、间接证据多、一对一证据多、书证多等特点。在司法实践中,对案件的认定和证据的使用还存在着轻信口供,草率定案等问题,这是一种主观主义办案作风,违背了实是求是的指导思想。     

试论中国社区警务的构建

从一定意义上讲,中国是实践社区警务最早的国家。构建中国社区警务是必要的,也是可行的。构建社区警务,必须贯彻改革精神,解放思想,转变传统思想观念,对警务重心、警务模式和警务风格重新调整,强化基层基础工作,深化派出所和刑侦体制改革,并应建立与之配套的民警巡逻体制。     

做好反腐倡廉工作是构建社会主义和谐社会的重要政治保证

构建社会主义和谐社会,把提高构建社会主义和谐社会的能力作为加强党的执政能力建设的重要内容,是党的十六大和十六届三中、四中全会提出的重大任务。社会主义和谐社会的基本特征是:民主法制、公平正义、诚信友爱、充满活力、安定有序、人与自然和谐相处。纪检监察工作和反腐倡廉工作是党的建设和国家政权建设的重要组成部分,提高纪监察能力和反腐倡廉水平是增强党的执政能力、巩固党的执政地位、完成党的历史使命的重要条件,对构建社会主义和谐社会有着重要作用。