青壮年猝死综合征SCN2B、SCN4B基因检测

目的寻找SCN2B、SCN4B基因的变异位点,探讨其与青壮年不明原因夜间睡眠中猝死(SMDS)的关系。方法提取SMDS病例组及健康对照组的基因组DNA,采用聚合酶链式反应(PCR)方法扩增SCN2B、SCN4B基因编码区外显子、外显子-内含子交界区以及3'侧翼区序列,直接行DNA测序以明确遗传变异类型。结果在病例组中共检测到4个变异位点,c.237+27AG,c.*38CT,c.174CT(p.C58C)和c.*7CT。结论本研究首次对中国人SMDS病例进行了SCN2B、SCN4B基因的检测,上述基因是否为中国人SMDS的易感基因尚有待进一步研究证实。     

过敏性休克死亡豚鼠心、气管、喉头IL-10表达的死后变化

目的研究过敏性休克死亡豚鼠心、气管、喉头白细胞介素10(Interleukin-10,IL-10)表达的死后变化规律,寻找过敏性休克死亡的法医学诊断的客观指标。方法参照文献建立豚鼠过敏性休克死亡动物模型,采用免疫组织化学的方法,应用图像分析技术,检测各组豚鼠心、气管、喉头IL-10在死后0h、48h(4℃冷藏)、5d(-20℃冷冻)、7d(-20℃冷冻)四个时间点的表达量。结果在健康的豚鼠(对照组)心肌细胞、气管、喉头粘膜上皮细胞中IL-10均呈弱阳性表达,而在过敏性休克死亡豚鼠(实验组)心肌细胞、气管、喉头粘膜上皮细胞中IL-10均呈较强阳性表达,各实验组与相应的对照组之间均有显著差异(p0.05);随着死后时间的延长,在死后冷藏48 h或冷冻7 d内,IL-10在心、气管、喉头的表达量呈缓慢减少趋势,但各实验组之间IL-10的表达量均无显著差异(p0.05)。结论死后冷藏48h或冷冻7d内尸体,其心肌细胞、气管、喉头粘膜上皮细胞IL-10呈较强阳性表达,这可能成为过敏性休克死亡法医学鉴定的较好的客观指标。     

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。