机械性窒息死亡他杀案件的法医学分析

目的分析机械性窒息死亡他杀案件的现场及尸检特点,为此类案件的法医学检验鉴定及案件侦破提供参考。方法采用分类描述性研究方法,对东莞市2009-2012年的152例机械性窒息死亡的他杀案件进行回顾性分析。结果机械性窒息死亡他杀案件的发生具有一定时间性、区域性。闹市区室内为高发场所,女性及儿童为高危人群。加害方式以徒手、扼颈为主。不同窒息征象间的检出率差异有统计学意义。被害人指甲拭子提取到案犯DNA成分几率高(21.05%)。结论机械性窒息致死的他杀案件具有区域性,高发人群较固定。窒息征象的检出对于窒息的死因认定具有法医学意义,被害人指甲拭子应作为DNA的重点检验部位。     

3种方法对临床菌血症样本细菌DNA提取的比较

目的寻找一种简单、有效的提取临床菌血症样本细菌DNA的方法．方法分别用碱液加热裂解法、溶菌酶法和商品DNA提取液法提取细菌DNA，用临床常见病原菌特异性引物，进行PCR扩增．结果（1）商品DNA提取液法提取革兰阴性菌DNA，经PCR扩增后得到特异性目的片段，但未扩增出革兰阳性菌的特异性目的片段；（2）碱液加热裂解法、溶菌酶法提取革兰阴、阳性菌DNA，经PCR扩增后均得到特异性目的片段，但碱液加热裂解法比溶菌酶法能扩增到更低浓度的细菌DNA目的片段．结论碱液加热裂解法是3种方法中最简单有效的细菌DNA提取方法，适用于临床病原菌基因组DNA提取．     

A review of DNA profiling success for laser microdissected forensic casework samples

ABSTRACT

Laser microdissection (LMD) is a tool that is used in forensic laboratories for the analysis of DNA from specifically targeted cells. Since 2010, the Institute of Environmental Science and Research Limited’s (ESR) Forensic Biology laboratory has applied LMD DNA testing to a variety of sexual assault casework samples where small numbers of sperm are present in cell mixtures. In this paper we review the DNA profiling results obtained from semen-stained casework samples that have been analysed using the LMD DNA methodology developed in our laboratory. Dissected sperm have been analysed using the AmpFISTR Identifiler amplification kit at 28 cycle PCR, the AmpFISTR MiniFiler amplification kit at 30 cycle PCR, or the AmpFISTR SGM Plus amplification kit at 34 cycle PCR, depending on the number of sperm recovered and on consideration of other circumstantial case information, such as the time since intercourse (TSI). From a review of these data, success rates for different sample numbers of sperm recovered from semen-stained samples are determined. The DNA profiling results obtained from three cases where laser microdissection has been used are also presented to demonstrate the success of the LMD testing strategy in a forensic laboratory.     

Would the real allele please stand up? Compiling DNA artefacts for the GlobalFiler PCR amplification kit – a South Australian approach

ABSTRACT

This article presents a compendium of DNA artefacts observed using the GlobalFiler and GlobalFiler Express PCR kits (ThermoFisher Scientific) during DNA reference profile assessment at Forensic Science SA (FSSA). The data are currently used to assist with the interpretation of GlobalFiler DNA evidence profiles encountered in the course of routine case work at FSSA. Over 1000 reference profiles have passed observation by reference DNA run readers. An artefact was considered confirmed if it was observed in five or more individual reference samples and could be confirmed upon re-PCR. Artefacts were documented in the following two categories: (1) those with a location independent of true allelic products and (2) those with a location relative to a true allelic product. The artefacts observed were positioned within and outside of allelic designations and were typically less than 1% of the closest allelic product. Artefacts reported here are observed from reference samples only. Their causes are largely unknown and warrant further investigation. Post-developmental artefacts may be encountered due to storage and handling conditions. As such, continued monitoring of reference samples for low level artefacts is warranted.     

High expression of human augmenter of liver regeneration in E. coli

Heat-stable hepatocyte stimulatory activity has been described in the liver of weanling rats and pigs. This growth factor is called hepatocyte stimulator substance (HSS)[1]. Hagiya et al[2]reported the complete amino acid sequence of a 30KDa band from their purified product of rat liver and the cloning and sequence analysis of its cDNA. They called it augmenter of liver regeneration (ALR). Our previous study[3]demonstrated that human ALR had been cloned and sequenced. To further study the bioactivity of human ALR (hALR), we constructed a highly expressed vector pBV-hALR in E. coli and about 20% of somatic protein of rhALR was expressed.     

精浆弹性硬蛋白酶与精子 DNA 完整性及精液参数的影响分析

目的：探讨男性不育患者精浆弹性硬蛋白酶水平与精子 DNA完整性及精液质量之间的关系。方法选择148例男性不育患者根据精浆弹性硬蛋白酶的检测结果将所有患者分为 A、B、C3组。 A组为精浆弹性硬蛋白酶含量＜290 ng／mL；B组为精浆弹性硬蛋白酶含量290～1000 ng／mL；C 组为精浆弹性硬蛋白酶含量＞1000ng／mL。用酶联免疫吸附试验（ ELISA）检测精浆中弹性硬蛋白酶，精子 DNA完整性检测用精子染色质扩散法（ SCD），精液参数用计算机辅助精子分析系统检测。结果 A组与 C 组相比精子存活率与前向运动精子百分率（PR）均升高、精子DNA碎片指数（DFI）明显降低，差异均具有统计学意义（ P＜0．05）。 B组的上述指标与A组相比，差异均无统计学意义（ P ＞0．05）。结论精浆弹性硬蛋白酶与精子DNA完整性及精液质量密切相关，生殖道感染是影响男性精液质量的一个重要原因。     

HBV微环DNA重组质粒的构建及初步药效学实验

目的 构建HBV的治疗性微环质粒并进行体内外实验研究. 方法 采用聚合酶链式反应扩增HBV包膜蛋白preS2.S抗原基因和hIL-2-IFNγ融合基因,将2个基因分别克隆于ZY781载体,得到亲本质粒preS2.S/ZY781和hIL-2-IFNγ/ZY781,测序正确后,用阿拉伯糖诱导生成微环质粒pMCS2.S和pMCIIF. 转染微环质粒于COS-7细胞株,用酶联免疫吸附法定量法检测24、48、72 h时目的蛋白的表达情况.双质粒联合在体电脉冲注射BALB/c小鼠,4周后处死小鼠,检测特异性的体液和细胞免疫应答. 结果 微环质粒pMCS2.S和pMCIIF转染COS-7细胞后,目的蛋白HBsAg、白细胞介素-2、干扰素 γ在不同时间均有表达,在48 h达峰值,分别为(60.3±7.8)、(17.7±1.9)、(10.3±0.9) ng/ml. 微环质粒pMCS2.S能诱导小鼠产生HBsAb的抗体保护,佐剂pMCIIF可显著增强其免疫效果,在低剂量5μg/只就能诱导较强特异性细胞和体液免疫. 结论 成功构建了HBV微环质粒pMCS2.S及其佐剂微环质粒pMCIIF,其有较好的体内外活性.     

高通量基因测序对血清学筛查临界风险孕妇的应用价值

目的：探讨对于血清学筛查临界风险的孕妇应用高通量基因测序价值。方法选择2012年5月至2015年5月在常州市妇幼保健院产前诊断中心就诊的1066例血清学筛查临界风险的孕妇，在知情同意的原则下抽取孕妇外周血，提取血浆中胎儿游离DNA ，制备文库，采用Illumina NextSeq500测序平台对其进行测序分析，对测序提示的染色体异常患者行羊膜腔穿刺，羊水细胞培养后染色体G显带核型分析。结果1066例样本中，高通量基因测序提示15例染色体非整倍体异常，经知情同意，13例孕妇自愿接受羊水产前诊断，其中7例羊水G带核型结果与测序结果一致，包括4例21三体，1例18三体，1例47，XXX ，1例47，XXY ，其余6例G带核型正常。结论高通量基因测序可作为血清学筛查临界风险孕妇的有效补充检测手段。