A real-time multiplex SNP melting assay to discriminate individuals

A method that quickly and inexpensively differentiates crime scene samples from multiple donors would expedite casework analysis by allowing the selection of probative items requiring comprehensive testing. This new method need not be perfectly definitive nor give a complete 13 locus short tandem repeat (STR) profile; it simply must be able to differentiate between most victim and suspect samples. We describe the development of multiplex, single nucleotide polymorphism (SNP), fluorescence resonance energy transfer-based real-time polymerase chain reaction (PCR) assays to fulfill this need. Dual probes, one fluorescently labeled and the other labeled with a quencher, are monitored during a melt analysis to reveal an increase in fluorescence, which allows the assessment of the two SNP alleles. Two alternate 6-plex assays (with and without gender determination) have been developed for the six-color RG6000 real-time instrument (Corbett Robotics, Inc.) and one seven SNP plus gender assay (performed as two 4-plex assays, one with gender the other without) have been developed for use in four/five color real-time instruments. This technique can discriminate between 95% and 99% of samples from different individuals. This assay is fast (approximately 2 h), much less expensive than STR analysis, and uses a real-time PCR instrument which is found in most forensic and molecular biology labs.     

大鼠急性心肌缺血后肌浆网RyR2 mRNA表达变化

目的 研究大鼠急性心肌缺血后心肌肌浆网兰尼碱受体蛋白2(ryanodine receptor 2,RyR2)mRNA表达的变化.方法 将SD大鼠分为正常对照组、心肌缺血组和缺血性猝死组.采用腹腔注射垂体后叶素的方法复制大鼠急性心肌缺血和猝死模型,对心肌进行半定量荧光RT-PCR检测,观察RyR2 mRNA表达水平的变化.结果 与正常对照组相比,不同时间和不同程度的急性心肌缺血后心肌肌浆网RyR2 mRNA表达均显著降低(P<0.05).结论 心肌缺血性损伤可诱导心肌钙调控蛋白RyR2 mRNA表达下调.     

Pig-mentation: postmortem iris color change in the eyes of Sus scrofa

Experienced forensic pathologists and examiners may be familiar with the phenomenon of postmortem iris color change; however, only Knight, Simpson's forensic medicine, Arnold, London, 1997; Ref. 1 and Saukko and Knight, Knight's forensic pathology, 3rd ed., Arnold, London, 2004; Ref. 2 have referred to it in the literature, and to date, there have been no published scientific research studies on this taphonomic artifact. A controlled experiment was conducted of postmortem changes to isolated Sus scrofa eyes. The eyes (n = 137) were separated into three groups and each sample was observed for 3-day postmortem at a different temperature. In addition, a Sus scrofa head was obtained to observe postmortem changes of eyes in situ. All isolated blue eyes in the experiment, at room temperature and higher, changed to brown/black within 48 h. The in situ blue eye, at room temperature, turned brown/black within 72 h. If iris color consistently changes postmortem in humans, then this taphonomic artifact must be incorporated into victim identification protocol, including disaster victim identification software, and autopsy reports to prevent inaccurate victim identification and inappropriate exclusion from the identification process.     

超高效液相色谱法比较鲜川芎和川芎药材中6种成分含量

目的 采用超高效液相色谱法(ultra-high performance liquid chromatography, UPLC)比较鲜川芎和川芎药材中6种活性成分(川芎嗪、阿魏酸、洋川芎内酯I、洋川芎内酯H、洋川芎内酯A、藁本内酯)的含量差异,为该药材的产地加工提供实验依据。方法 采用75%甲醇超声提取,UPLC测定鲜川芎和川芎药材中6种活性成分,根据样品处理条件计算质量分数并进行比较。流动相为乙腈(A)-0.1%甲酸水溶液(B),梯度洗脱,流速为0.2 mL/min,检测波长为294 nm,柱温30 ℃,进样体积2 μL。结果 经检测,阿魏酸、藁本内酯、洋川芎内酯A为鲜川芎和川芎药材的共有成分,鲜川芎中未检测到洋川芎内酯I及洋川芎内酯H。鲜川芎样品中各成分的质量分数为阿魏酸0.003 9%、洋川芎内酯A 0.006 4%、藁本内酯0.54%;川芎药材中各成分的质量分数为阿魏酸0.088%、洋川芎内酯I 0.20%、洋川芎内酯H 0.031%、洋川芎内酯A 0.76%、藁本内酯 1.3%。结论 川芎药材与鲜川芎中化学成分存在差异,推测在川芎的产地加工及贮藏过程中存在成分的转化。     

超高效液相色谱-四级杆飞行时间串联质谱快速分析复方守宫散化学成分及血中移行成分

目的 采用超高效液相色谱-四级杆飞行时间质谱(ultra-performance liquid chromatogra-phy/quadrupole time-of-flight-tandem mass spectrometry,UPLC-Q-TOF/MS)对复方守宫散中化学成分及其血中移行成分进行定性分析,为该制剂的...     

湘黄精中高异黄酮类化学成分研究

目的 对黄精属植物湘黄精Polygonatum hunanense H. H. Liu&B. Z. Wang的干燥根茎进行化学成分研究。方法 综合运用硅胶、Sephadex LH-20凝胶、半制备HPLC等方法对湘黄精乙酸乙酯部位进行分离纯化,根据波谱数据、理化性质等技术手段对化合物结构进行鉴定。结果 最终从湘黄精根茎95%乙醇提取物中分离得到13个化合物,分别鉴定为5,7-dihydroxy-6-methyl-3-(4-methoxybenzyl)-chroman-4-one (1)、5,7-dihydroxy-3-(2-hydroxy-4-methoxybenzyl)-chroman-4-one (2)、ophiopogonanone G (3)、3-(4′-hydroxy-benzyl)-5,7-dihydroxy-6-methyl-chroman-4-one (4)、(±)-5,7-dihydroxy-6,8-dimethyl-3-(2′-hydroxy-4′-methoxybenzyl-)chroman-4-one (5)、3-(2′,4′-dihydroxybenz...     

颜色的中西文化含义差异比较

由于世界上各民族在文化风俗、地理位置、历史传统、宗教信仰、民族心理、思维习惯、意识形态等方面所存在的差异,同一颜色词语有时表现出明显的语义差异,刻上深深的民族文化烙印,甚至构成了人们对经过引申、转义之后颜色的崇尚或禁忌。本文旨在通过探讨不同文化之间颜色象征意义的差异,指出这种差异是在各民族文化历史背景的审美心理差异基础上发展衍生的,是在社会发展和历史沉淀中约定俗成的,是一种文化现象。对这种文化现象的解析有助于不同文化间的相互交流的发展,顺利地进行跨文化交际。     

ABO基因分型及其在法医学中的应用

为建立一种ABO血型系统基因分型方法，采用PCR-RFLP技术，成功地将ABO系统区分为AA，AO，AB，OO，BB，BO六种基因型。对240名中国汉族无关个体血样的ABO（基因型频率调查结果表明，6种基因型的频率分布为0．0125～0．3834，符合Hardy－Weinbeng遗传平衡法则（P＞0．1），其DP值为0．8161。家系分析表明，亲代a、b、o基因传递遵守孟德尔遗传规律。对法医学中常见的血痕、混合斑、骨组织及毛发根部等生物样品进行检测，均能准确判定ABO基因型，并可在实际案件鉴定中应用。     

浙江汉族人群6个STR基因座的遗传多态性的调查

目的进一步完善浙江省汉族人群STR基因座遗传多态性的调查,为其应用提供基础数据。方法采用AmpF1STRSGMplus和AmpF1STRCoficer反应试剂盒,使用ABI310型基因分析仪对浙江汉族人群200名无关个体血样进行了D16S539、D2S1338、D19S433、TH01、TPOX和CSF1PO6个STR基因座遗传学分析。结果分别发现了9、15、15、11、8、10个等位基因,发现的基因型分别为23、42、35、19、16、17个,其分布经X2检验均符合Hardy-Weinberg平衡定律,并分别统计了6个STR基因座的H、DP、PM、PE及PIC参数。结论6个STR基因座适合法医学应用。