溴氰菊酯在急性中毒大鼠体内的分布

目的建立溴氰菊酯灌胃急性中毒大鼠模型及生物样品中溴氰菊酯的气相色谱-电子捕获检测器法(gas chromatography-electron capture detector,GC-ECD),研究溴氰菊酯在急性中毒大鼠体内的分布特征,为溴氰菊酯中毒案件的法医学鉴定提供参考依据。方法不同剂量(512和1024mg/kg)溴氰菊酯灌胃染毒大鼠后1.5 h处死,迅速解剖取血液、心、肝、肺、肾、脑等,样品经无水硫酸钠研磨,混合溶剂[V(石油醚)∶V(丙酮)=4∶1]浸提,GC-ECD法定量检测溴氰菊酯含量。结果溴氰菊酯与内源性杂质分离良好,血液和肝中溴氰菊酯定量限分别为0.1μg/mL和0.1μg/g(S/N≥10),血液中溴氰菊酯的回收率为91.55%~134.37%,日内精密度和日间精密度均小于5.67%。溴氰菊酯在512 mg/kg剂量灌胃染毒大鼠体内分布为:肺肝心肾血液脑;在1 024 mg/kg剂量灌胃染毒大鼠体内分布为:肺血液心肾脑肝(P0.05)。结论本研究建立的溴氰菊酯GC-ECD检测方法灵敏度高。溴氰菊酯在体内的分布存在剂量依赖,血液、心、肝、肺、肾、脑可作为体内溴氰菊酯分析的良好检材。     

试析俄东部地区民俗文化旅游的开发

俄东部地区少数民族众多，民俗风情古朴原始，民族传统文化完整，民俗旅游资源丰富。依托其丰富的民俗资源开发具有民俗文化特色的旅游项目，是发展民俗文化旅游、开展区域文化旅游合作和带动区域旅游经济跨越式发展的有效途径。     

The Optimization of Electrophoresis on a Glass Microfluidic Chip and its Application in Forensic Science

Microfluidic chips offer significant speed, cost, and sensitivity advantages, but numerous parameters must be optimized to provide microchip electrophoresis detection. Experiments were conducted to study the factors, including sieving matrices (the concentration and type), surface modification, analysis temperature, and electric field strengths, which all impact the effectiveness of microchip electrophoresis detection of DNA samples. Our results showed that the best resolution for ssDNA was observed using 4.5% w/v (7 M urea) lab‐fabricated LPA gel, dynamic wall coating of the microchannel, electrophoresis temperatures between 55 and 60°C, and electrical fields between 350 and 450 V/cm on the microchip‐based capillary electrophoresis (μCE) system. One base‐pair resolution could be achieved in the 19‐cm‐length microchannel. Furthermore, both 9947A standard genomic DNA and DNA extracted from blood spots were demonstrated to be successfully separated with well‐resolved DNA peaks in 8 min. Therefore, the microchip electrophoresis system demonstrated good potential for rapid forensic DNA analysis.     

The Diagnostic Value of Quantitative Assessment of Diatom Test for Drowning: An Analysis of 128 Water‐related Death Cases using Microwave Digestion‐Vacuum Filtration‐Automated Scanning Electron Microscopy

The value of diatom test for the diagnosis of drowning remains controversial. The conventional forensic diatom test with low sensitivity is not a useful tool to provide accurate information about diatom in the tissues and organs. To improve the sensitivity of the diatom test, we developed a novel method called the Microwave Digestion‐Vacuum Filtration‐Automated Scanning Electron Microscopy (MD‐VF‐Auto SEM) method which resulted in a high recovery of diatoms. In this article, we analyzed 128 water‐related death cases. Our results showed that the MD‐VF‐Auto SEM method could achieve a much higher positive rate (0.97) in drowning cases. Large amounts of diatoms are retained, even concentrated, in the lung tissues during the intense breathing movement in drowning process. This might be useful for the diagnosis of drowning. Our research indicates that the MD‐VF‐Auto SEM method would be a valuable methodology in the study of diatom test for the forensic community.     

THE INNOVATION OF LAW: FROM CHINESE PERSPECTIVE

It is a new and meaningful research in discovering the impact of technology on law, and the reciprocal relationship between law and invention. This paper starts with a Chinese ancient tragedy to disclose the direct relationship between technology and law that was reflected in the play. It explains the “Needham Puzzle” that was engraved in Chinese law in discussion with the indirect impact of technology on law in the Chinese legal system. After that, the Chinese patent system is analyzed specifically to discover the law of innovation. It also discusses the impact of law on technology in the field of patent system.     

现代公益事业法人治理结构研究

公益事业单位既是中国事业单位集群的主体,也是分类改革后能够继续保留在国家事业序列内的唯一类型。现有公益事业单位虽然在职能目标上符合国家事业的要求,但在组织形态上还需要进一步转变成为"现代公益事业组织"。在法人自治基础上的科学管理是现代公益事业组织的基本要求和基本特征,公益事业单位内部治理模式改革的核心是构建与组织属性相符合的法人治理结构。公益事业单位由于其职能范围的广泛性、组织地位的独立性以及行为目标的公共性,适宜采用"理事会+执行负责人"的法人治理结构,同时还应建立一个由利益相关人广泛参与的共同治理架构。     

猪瘟病毒石门株E2蛋白的单抗制备及抗原表位鉴定

用猪瘟病毒石门株E2(SM-E2)的BC区原核表达蛋白为抗原免疫BALB/c小鼠,应用杂交瘤技术制备单抗,测定其效价并鉴定抗原表位。结果表明,制备了1株持续、稳定分泌抗SM-E2的单克隆抗体2B10,效价在1∶600～1∶1 000之间。单抗2B10能与常规SDS-PAGE转膜的SM-E2蛋白发生反应,表明其识别的是线性表位,并且该单抗只能识别石门毒株,而不能与C株疫苗及2型流行毒株发生反应。点突变E2蛋白ELISA分析结果表明,该单抗不与707、709、711、713和714位突变蛋白发生反应,表明单抗2B10识别的线性表位为707I-X-P-X-G-X-G-G714。该单抗对猪瘟病毒抗原多样性分析和致病机制研究有一定的应用价值。     

鸡朊蛋白的原核表达及其抗血清的制备

运用分子生物学方法分别构建了pGEX-4T-ChPrP1、pGEX-4T-ChPrP2、pGEX-4T-ChPrP3重组表达载体,经转化大肠杆菌BL21(DE3)和IPTG诱导表达,并对融合蛋白诱导表达时间、IPTG浓度进行优化,诱导表达的GST-ChPrP(25～247)融合蛋白经GST-trap FF柱亲和层析纯化,用纯化后的GST-ChPrP(25～247)融合蛋白免疫新西兰雄兔,以探究鸡朊蛋白的结构与功能。结果表明,本试验成功构建了pGEX-4T-ChPrP1、pGEX-4T-ChPrP2、pGEX-4T-ChPrP3重组表达菌,SDS-PAGE分析结果显示,得到大小为50、42和32ku左右的融合蛋白GST-ChPrP(25～247)、GST-ChPrP(96～247)、GST-ChPrP(174～247),与预期结果基本相符,在最适条件下每升pGEX-4T-ChPrP1菌液通过GST-trap FF层析柱获得的GST-ChPrP(25～247)融合蛋白达10mg,制备的抗血清经琼脂糖凝胶扩散试验检测到较高的多克隆抗体效价(1∶32);Western-blot分析结果显示,融合蛋白GST-ChPrP(25～247)、GST-ChPrP(96～247)和GST-ChPrP(174～247)及鸡脑组织蛋白均可与该抗血清发生特异性免疫反应。证实,鸡重组朊蛋白具有良好的抗原性,与鸡脑组织朊蛋白具有相同的抗原成分。     

鸭病毒性肝炎病毒RNA聚合酶在真核细胞中的表达及亚细胞定位

为研究鸭病毒性肝炎病毒RNA依赖性RNA聚合酶的分子生物学特性及其在细胞中的定位,构建了RNA依赖性RNA聚合酶与增强型绿色荧光蛋白融合表达的真核重组表达质粒;然后用脂质体介导转染MDCK细胞。分别使用荧光显微镜、激光共聚焦显微镜以及4,6-二脒基-2-苯基吲哚(DAPI)染色和蛋白免疫电泳试验等方法检测了RNA依赖性RNA聚合酶在细胞中的表达和亚细胞定位情况。结果表明,RNA依赖性RNA聚合酶能与增强型绿色荧光蛋白一起在MDCK细胞中良好表达,且主要分布于细胞核中,Western-blot分析结果显示,融合蛋白在80ku处出现阳性条带,说明表达的外源蛋白具有免疫活性。表明成功构建了RNA依赖性RNA聚合酶与增强型绿色荧光蛋白融合表达载体,并在真核细胞内实现了表达,通过检测荧光蛋白,发现RNA依赖性RNA聚合酶主要在细胞核中发挥复制酶的功能。     

保证期间的困扰--兼论法律与司法解释的关系

保证期间既可以由当事人约定,也可以在无约定时由法律规定,因此其既不是诉讼时效期间,也并非除斥期间,而仅应视为是保证人承担保证责任或履行保证合同项下代偿义务的期限。对当下保证期间(限)制度的考察显示,司法解释仍具有超越上位法的冲动和纪录,从而使得既有的司法实践经验即便在法律出现大幅度修改的情况下,仍可通过司法解释得以保存和延续。而将上级法院观点预先成文化的做法,恰与严格限制法官个人在个案中诠释法律的惯行构成鲜明对比。从2011年12月开始,最高人民法院不定期发布指导性案例。但若想以其取代相沿已久的司法解释制度,恐怕还有待时日。