Development of Multiple Assays using 46 SNPs for Comprehensive mtDNA Haplogrouping and Application to Highly Degraded DNA

Six multiplex PCR systems using single‐base extension reactions to analyze 46 mitochondrial DNA (mtDNA)‐coding region single nucleotide polymorphisms (SNPs) that define 42 haplogroups, that is, 24 major mtDNA haplogroups and 18 subclades, were devised. To improve the usefulness of the established systems for the analysis of degraded DNA samples, novel primers to render amplicons with sizes <150 bp were designed. By applying these systems to 214 Japanese individuals, 24 different haplogroups (power of discrimination = 93.4%) were found. To assess the effectiveness of our systems in grouping degraded DNA, an ancient bone sample of a Jomon skeleton was analyzed and then classified as haplogroup N9b. We conclude that the present systems are powerful screening tools for major haplogroups of mtDNA in addition to the prevalent subhaplogroups in the Japanese population and that these systems are capable of analyzing highly degraded DNA samples in forensic studies.     

分泌抗人精液特异蛋白P_(30)单克隆抗体杂交瘤细胞系的建立和初步应用

作者通过杂交瘤技术建立了9株产生抗精浆特异蛋白 P_(30) 单克隆抗体的杂交瘤细胞系。它们是由 SP2/0骨髓瘤与经 P_(30) 免疫的 BALB/C 小鼠脾细胞按常规方法进行细胞融合、并经克隆化筛选得出.这些细胞株均经体外培养3个月以上能够稳定分泌抗 P_(30) 单克隆抗体。该抗体只能识别纯化的 P_(30) 和精液中的 P_(30) ;与人精液以外的其他体液和多种人体组织无交叉反应;与几种常见动物的精液和血液无交叉反应。这些 P_(30) 单克隆抗体均属 IgG 类和 IgG_1亚类。其培养上清液和腹水的抗体效价最高分别达到320和128,000。以 ELISA 法应用这些单克隆抗体能很好地鉴定精液和精斑。     

A New Method for Human ABO Genotyping Using a Universal Reporter Primer System

Abstract: We developed a new method for forensic ABO genotyping based on a universal reporter primer (URP) system. This allows for the simultaneous detection of six single nucleotide polymorphism (SNP) sites in the ABO gene (nucleotide positions 261, 297, 526, 703, 796, and 803). This URP system provides obvious peaks, ranging from 82 to 151 bp in length. ABO genotypes were classified and successfully genotyped by our method, including minor alleles that may cause a discrepancy between the genetic data and serological phenotypes. Full profiles were identified using as little as 0.1 ng (0.05 ng/reaction) of standard K562 and 9947A DNA. Moreover, the success rate of genotyping from a URP system was much higher than that from a conventional primer extension method in degraded DNA. This method enables simple and rapid detection of multiple SNP sites on human ABO genes and is highly specific and sensitive when using limited and degraded DNA.     

微量体表脱落上皮细胞的DNA检验

目的　建立微量体表脱落上皮细胞的DNA检验方法。　方法　采用Chelex -10 0法提取DNA ,以Microcon -10 0纯化柱纯化浓缩DNA ,用ProfilerPlus试剂盒PCR扩增后用 3 10基因分析仪检测。　结果　 10种常见粘附有体表脱落上皮细胞的样本 10 0个 ,其中 7种 70个样本成功检测到 10个STR位点的分型 ,检出率为 10 0 % ,其余 3种样本检出率分别为 5 0 %、2 0 %、10 % ,将该方法应用于 2例实际检案 ,取得满意效果。　结论　所建立方法稳定可靠 ,易于操作 ,适用于多种检材 ,为微量体表脱落上皮细胞的DNA检验提供了确实可行的检验方法     

我国刑法人身权保护现状和问题

人身权利是作为社会主体的人赖以生存和发展的基础,是现代人权和有关法律规范保护的重要内容.完整分析我国刑法的人身权概念和范围,详细总结我国参加和承认的国际人权公约中人身权保护的一般国际标准,深入说明我国刑法在人身权各方面保护的基本状况和存在的主要问题,提出根据国际人权标准提高我国人身权保护水平的方向性思考,指出降低起刑点才是改革中国刑事法律制度以及法律制度的正确方向.     

人权基本概念的演变与人权发展历程

人权是我们这个时代的共同话语,同时也是一个深刻、敏感并极富争议性的论题,其中人权概念就是一个必须认真对待的基本问题。现代人权概念是一个历时性与共时性相互作用的产物。在人类的人权发展史上,曾先后出现过人权的几个基本概念,厘清这些概念将有助于我们理解人权概念本身的演变及其内涵与外延的丰富与发展过程。它们从某个角度记载了人权事业发展的曲折艰难历程和人类在人权事业上不断进步的足迹。     

Evaluation of Commercial Kits for Dual Extraction of DNA and RNA from Human Body Fluids, ,

STR typing of DNA evidence can identify the donor with a high power of discrimination but cannot identify the tissue origin of a body‐fluid stain. Using RNA to attribute a crime scene stain to a particular tissue may aid in reconstruction efforts. With blood from 10 donors, four DNA and RNA coextraction kits were evaluated by measuring yields and STR and mRNA profiles. T tests indicated some significant differences in kit performance. The Zymo Research ZR‐Duet^? kit performed best based on average DNA (41.4 ng) and mRNA (4.07 ng) yields and was the only kit to provide complete DNA/RNA profiles for all samples. The consistency of this kit was challenged by data from additional blood and saliva donors. Further testing is advised before a superior kit is unequivocally chosen. Stand‐alone DNA or RNA purification generally offers higher yield, but coextraction may still allow successful STR profiling and tissue source identification.     

Modeling Bone Surface Morphology: A Fully Quantitative Method for Age‐at‐Death Estimation Using the Pubic Symphysis

The pubic symphysis is widely used in age estimation for the adult skeleton. Standard practice requires the visual comparison of surface morphology against criteria representing predefined phases and the estimation of case‐specific age from an age range associated with the chosen phase. Known problems of method and observer error necessitate alternative tools to quantify age‐related change in pubic morphology. This paper presents an objective, fully quantitative method for estimating age‐at‐death from the skeleton, which exploits a variance‐based score of surface complexity computed from vertices obtained from a scanner sampling the pubic symphysis. For laser scans from 41 modern American male skeletons, this method produces results that are significantly associated with known age‐at‐death (RMSE = 17.15 years). Chronological age is predicted, therefore, equally well, if not, better, with this robust, objective, and fully quantitative method than with prevailing phase‐aging systems. This method contributes to forensic casework by responding to medico‐legal expectations for evidence standards.     

运用二代测序技术拆分混合STR分型

目的探讨二代测序技术在混合STR分型拆分中的应用。方法对一例强制猥亵妇女案中受害人颈部的拭子和血样作DNA提取,以Precision ID GlobalFiler^TM NGS STR Panel v2试剂盒制备文库,经Ion S5测序仪测序,运用Torrent_Suite_v5.2.1软件进行数据分析后,将检测基因座的序列多态STR分型与长度多态STR分型进行比较。结果在D8S1179、D21S11、D2S441、D2S1338、D10S1248五个基因座发现存在序列特异的等位基因亚型,利用这些亚型对混合STR分型进行了成功拆分。结论二代测序技术提供的等位基因序列信息可对混合STR分型的拆分起到帮助作用。