Simplified Direct PCR Method for Reference Buccal Samples Using a Non-FTA Card by Omitting the Pretreatment Step

When using non-FTA cards in commercial multiplex STR kits for direct PCR, pretreatment steps with specific buffers are recommended. Here, we designed a rapid direct PCR method utilizing a non-FTA card, Oral Cell Sampling Kit, by omitting the pretreatment step involving Prep-n-Go™ Buffer, and it showed compatibility with the GlobalFiler™ Express PCR Amplification Kit, GlobalFiler™ PCR Amplification Kit, and PowerPlex^® Fusion system. To optimize the PCR conditions, we tested the method with different final PCR volumes and cycles. Finally, we conducted a performance test using 50 Korean buccal samples and confirmed the high performance of the method, detecting more than 90% of the samples with full profiles when using GlobalFiler™ PCR Amplification Kit and PowerPlex^® Fusion system at 29 cycles in a 10 μL final PCR volume. Thus, we report a simple direct PCR set-up to analyze reference samples collected using a non-FTA card manufactured in Korea.     

中国高校教师聘任制政策文本研究

本文通过内容分析的研究方法，对依据“可获得性”原则搜集到的47所高等学校教师聘任制度样本，从13个方面比较了具体内容的异同，对我国目前各地高校教师聘任制度改革的整体状况进行了统计分析和说明。研究发现，各地高校在政策制定上都与国家相关规定的基本精神保持了高度一致，相互之间不存在显著差异，这些都充分反映了我国高等教育高度行政化、高度集权化的特点。现阶段，我国高校教师聘任制度建设已初步形成体系，并发挥了一定作用，但是仍然存在各个方面的具体问题，需要进一步深化和落实聘任制度改革。     

Successful DNA typing of ultrafiltered urines used to detect EPO doping

Endogenous and exogenous erythropoietin (EPO) present in urine can be distinguished according to their isoelectric profiles. This methodology requires urine samples to be concentrated about 200 to 1000 times with manipulations that should remove most of the cells occurring in the original sample. In this study, we tried to obtain DNA profiles from 10 ultrafiltered urines (retentates) in order to evaluate whether a formal genetic identification was technically feasible. No nuclear DNA profiles could be established from retentates, despite 34 PCR-cycles amplifications. Contrastingly, mitochondrial DNA (mtDNA) profiles were obtained for 9 out of the 10 retentates. Apart from some particularities, retentate mtDNA profiles were all distinct and matched mtDNA profiles of corresponding reference samples.     

Implementation of a robotized real-time PCR setup for the use of the Quantifiler™ Human DNA Quantification Kit

Human DNA quantification occupies a central role within the DNA analytic process of forensic casework samples as DNA quantification results have an important impact on the quality of the short tandem repeat data. Manual processing for the setup of quantification reactions can be time consuming and labor intensive. Therefore automation of quantitative real-time PCR setup was an important component of our DNA-analysis automation concept. Here we show the implementation of a robotized setup for the Quantifiler™ Human DNA Quantification Kit.     

心肌及传导组织内3种蛋白的变化与SMDS的相关性

目的 观察心肌及传导组织内肌动蛋白、血浆白蛋白及纤维连接蛋白的染色变化，探讨青壮年猝死综合征(SMDS)死后诊断的新方法。方法 应用免疫组化S-P法，对SMDS及冠心病猝死者心肌及传导系统内肌动蛋白、血浆白蛋白和纤维连接蛋白的进行染色观察。结果 22例SMDS例猝死者心肌及传导系统内的肌动蛋白缺染17例；血浆白蛋白染色阳性18例；纤维连接蛋白染色阳性15例。并发现在SMDS中，8例有CCS严重病变。结论 心肌及传导系统内血浆蛋白、肌动蛋白改变与SMDS密切相关，部分SMDS病例死前存在早期心肌缺血或梗死的改变。     

中毒鱼体材料中五氯酚的检验

目的　研究五氯酚的检验方法并对一起五氯酚钠投毒案进行检验。方法　采用高效液相色谱法及气相色谱 质谱联用法对案件所涉及的各种检验材料 ,尤其是鱼体材料中的五氯酚进行检验 ,同时分析新建技术的特点。结果　运用所建方法检出检材中的五氯酚。结论　所建方法可进行五氯酚钠投毒案件的检验 ,且适用范围广。     

The typing of group-specific component (Gc protein) in human blood stains

It is known that the typing of group-specific component (Gc protein) in human blood stains is difficult since Gc protein of the extracts of blood stains migrates more anodally to the α₁-globulin region in agar-gel immunoelectrophoresis, while Gc protein in liquid blood normally migrates to the α₂-globulin region. We have reported that the Gc protein found in the α₁-region is the result of binding of actin to Gc protein (Shinomiya, K., Kimura, H., Yoshida, K., and Shinomiya, T., J. Biochem., 92 (1982) 1163–1171, which renders it difficult to determine the Gc-phenotypes in the blood stains. On the basis of the above findings, we developed the method of phenotyping the Gc protein of human blood stains by agar-gel immunoelectrophoresis. Since the binding activity of actin to Gc protein is lost after treatment with a high concentration of guanidine HCl, the extracts of blood stains were treated with 4 M guanidine HCl to dissociate Gc protein and actin and then dialyzed to remove guanidine HCl. By this method we are able to determine the phenotypes of Gc protein in blood stains. The method we have developed is a useful tool in the forensic laboratory.     

气相色谱法同时测定血清中甲醇、乙醇、正丙醇

目的建立气相色谱同时测定血清中甲醇、乙醇、正丙醇含量的方法。方法改变气相色谱条件,以异戊醇为内标,采用气相色谱一氢火焰离子化检测器对血清直接进行检测。并通过待测组分与内标物的响应值比进行定量。结果GC／FID法检测血清中的甲醇、乙醇、正丙醇含量,得到了良好的线性关系。乙醇浓度从1～100mg／100ml。的线性关系式为Y=0．4145X＋0．0232（R2=0．9974）、浓度从100～1000mg／100mL的线性关系式为Y=0．4511X＋0．0746（R2=0．9911）,甲醇浓度从l-200mg／100mL的线性关系式为Y=0．2778X＋0．0493（R2=0．9983）。结论该方法操作简便快速,重现性好,通过检测正丙醇还可以推断腐败血样自身产生的乙醇量,是一种较为理想的血醇检测方法。     

超滤管在陈旧生物检材中的应用

目的探讨millipore超滤管滤过方法对陈旧生物检材DNA分型检验的应用价值。方法将23份陈旧血样分别剪取同样合适大小的血片3组,标为A、B、C组,磁珠法提取DNA,分别用80μL、80μL、20μL洗脱液洗脱得到模板DNA,其中A、C组模板直接扩增,B组用millipore超滤管滤过浓缩后扩增。PCR产物用AB3130x L基因分析仪检测,Gene Mapper ID V3.2软件进行自动分型。所得实验数据用SPSS软件分析处理。结果 A组没有1例样本扩出全部STR基因座,B组有18例样本扩出全部STR基因座,C组有11个样本扩出全部STR基因座。结论应用millipore超滤管滤过方法可以明显提高陈旧生物检材的DNA分型成功率。     

单细胞检验技术用于混合上皮细胞的分离和检验

目的建立单细胞显微捕获联合低体积扩增技术,用于混合上皮细胞检材分离检验。方法取5名男性口腔上皮细胞拭子浸泡液30μL,分别滴加到5份含同一女性皮肤表皮细胞拭子上,制成5份混合上皮细胞样本为实验组,同时制备同样的5份样本为对照组。实验组样本采用显微捕获单个口腔上皮细胞,并使用低体积扩增技术进行扩增;对照组用M48纯化试剂盒提取DNA,Identifiler试剂盒复合扩增,扩增体系为10μL。所有产物均用ABI 3130遗传分析仪进行STR分型。结果 5份实验组样本均得到男性STR分型结果,5份对照组样本则均仅得到混合分型结果。将该方法应用于1例强奸杀人案例检验,取得了满意效果。结论单细胞显微捕获联合低体积扩增技术可用于混合上皮细胞样本的分离检验。