基于HPLC-TOF-MS代谢组学方法的溴鼠灵毒性作用评价

目的分析溴鼠灵中毒大鼠尿液的代谢特征,揭示溴鼠灵干预对大鼠毒性作用的分子机制。方法通过构建大鼠溴鼠灵中毒模型,采用高效液相色谱-飞行时间质谱(high performance liquid chromatographytime of flight mass spectrometry,HPLC-TOF-MS)获取大鼠尿液代谢轮廓,并用正交偏最小二乘法-判别分析(orthogonal partial least squares-discrimination analysis,OPLS-DA)进行多变量统计分析,找出与溴鼠灵毒性作用密切相关的差异代谢物。结果 OPLS-DA得分图显示给药前后不同时间的大鼠尿液样本代谢物轨迹在各时间段内相似度较好,呈现各自聚类现象。比较溴鼠灵给药前后大鼠尿液样本,筛选出22个与溴鼠灵毒性相关的差异代谢物。结论溴鼠灵主要通过干扰大鼠体内的三羧酸循环、糖酵解、鞘脂代谢和色氨酸代谢等代谢通路发挥毒性作用,且溴鼠灵毒性作用具有累积效应。基于尿液HPLC-TOF-MS代谢组学方法可为溴鼠灵毒性作用的分子机制研究提供新思路。     

基于模式识别方法研究深静脉血栓形成大鼠尿液代谢物谱

目的采用代谢组学技术研究深静脉血栓形成(deep venous thrombosis,DVT)大鼠尿液中代谢物谱,筛选出可用于DVT诊断和法医学鉴定的小分子生物标志物。方法建立下腔静脉完全结扎DVT大鼠模型,分为模型组、假手术组和对照组,每组各10只。模型组和假手术组大鼠在建模后48 h于代谢笼中收集24 h尿液,同时对照组收集24 h尿液。核磁共振检测其代谢物谱,SIMCA-P 14.1软件进行模式识别,通过正交偏最小二乘法-判别分析(orthogonal PLS-DA,OPLS-DA)模型中的变量权重重要性排序(variable importance in projection,VIP)值结合Mann-Whitney U检验,寻找尿液中差异代谢物。结果模型组、假手术组和对照组大鼠尿液的代谢轮廓呈现显著性差异。通过偏最小二乘法-判别分析(partial least squares method-discriminant analysis,PLS-DA)模型可以有效判别模型组、假手术组与对照组。与对照组大鼠相比,DVT大鼠尿液中亮氨酸、谷氨酰胺、肌酸、肌酐和蔗糖水平上调,3-羟基丁酸、乳酸、丙酮、α-酮戊二酸、柠檬酸和马尿酸水平下调。结论 DVT大鼠尿液中的差异代谢物有望成为该疾病的候选生物标志物,该结果可为DVT的诊断、治疗和法医学鉴定提供研究基础。     

甲基苯丙胺亚急性中毒大鼠血清和尿液代谢组学研究

目的运用核磁共振研究甲基苯丙胺(MA)亚急性中毒大鼠血清和尿液代谢物变化,寻找可能的生物标志物及其代谢通路,为MA中毒及滥用的毒理机制提供思路。方法 16只SD大鼠随机分为亚急性中毒组和对照组,以腹腔注射MA建立亚急性中毒模型。运用1H-NMR结合正交偏最小二乘法-判别方法分析MA中毒大鼠与对照组的血清和尿液。结果以VIP1和P0.05筛选出7种差异代谢物分别为2-酮戊二酸、醋酸盐、甲胺、肌酐、丙酸、琥珀酸和甘氨酸,尿液中2-酮戊二酸、甲胺、肌酐、甘氨酸呈上升趋势且具有统计学差异(P0.05)。与差异代谢物相关的代谢通路为天冬氨酸、丙氨酸和谷氨酸代谢和柠檬酸循环。结论 2-酮戊二酸、甲胺、肌酐、甘氨酸可作为MA亚急性中毒的潜在生物标志物,天冬氨酸、丙氨酸和谷氨酸代谢和柠檬酸循环代谢途径参与MA亚急性中毒代谢过程,为MA中毒机制及鉴定MA滥用提供新思路。     

氰化物急性中毒大鼠的血浆代谢组学研究

目的 利用代谢组学技术研究急性氰化物中毒大鼠血浆中小分子代谢物的变化，寻找差异代谢物，分析与氰化物中毒相关的代谢途径，进而研究氰化物中毒的机制。方法 将健康SD大鼠随机分为空白对照组和实验组（雄性，每组5只），实验组灌胃3.2 mg/kg(1/2LD50)氰化钾溶液建立氰化物急性中毒模型，对照组灌胃等剂纯水，灌胃后分别于45 min,24 h,120 h眼眶静脉采血，通过高效液相色谱-飞行时间质谱采集大鼠血浆代谢谱，根据主成分分析（PCA）、正交偏最小二乘判别分析（OPLS-DA）以及t检验和变异倍数分析筛选差异代谢物，并通过KEGG(Kyoto Encyclopedia of Genes and Genomes)数据库进行代谢通路分析。结果 筛选出19个与氰化物中毒相关的差异代谢物，且随时间变化差异代谢物的种类存在差异，主要涉及胆汁分泌、氨基酸代谢等共11条代谢通路。结论 可将牛磺胆酸盐和牛磺鹅胆酸盐作为氰化物中毒潜在生物标志物，马尿酸可以作为两者的辅助指标，也可根据差异代谢物的不同进行服药时间的初步推断。氰化物会对机体酶的活性和能量代谢过程产生影响，且可能造成肝脏损伤。     

基于代谢组学技术的毒鼠强中毒机制研究

目的 研究毒鼠强中毒后大鼠的血浆代谢组学随时间变化规律，从代谢组学角度揭示其毒理学作用机制，为毒鼠强中毒的相关法医学研究提供依据。方法 将SD大鼠随机分成对照组和实验组(n=8)，实验组大鼠灌胃1/4LD₅₀(0.05 mg/kg)毒鼠强乙酸乙酯溶液，对照组灌胃等剂量乙酸乙酯溶液，灌胃后于24 h、5 d、16 d眼眶静脉采血(约1 mL)，应用超高效液相色谱-飞行时间质谱联用技术检测血浆中的内源性小分子物质。采用主成分分析(PCA)、正交偏最小二乘判别分析(OPLS-DA)等筛选显著差异代谢物，寻找与毒鼠强中毒相关的代谢通路。结果本研究以变量权重重要性排序(VIP)值>1和student t检验的P值<0.05为筛选标准，中毒后不同时间点共筛选出显著差异代谢物30种，包括鞘脂类、氨基酸类和脂肪酸类化合物，涉及到鞘脂信号通路、鞘脂代谢和ABC蛋白转运等10条代谢通路。结论 毒鼠强中毒后机体多条代谢通路受到显著影响，主要是不同程度的神经系统调节紊乱和能量代谢紊乱，整体变化呈现出累积效应，可为研究毒鼠强毒性作用机制提供依据。     

基于GC-MS研究百草枯中毒模型小鼠血清代谢产物变化

目的基于气相色谱-质谱联用技术(GC-MS)分析百草枯中毒模型小鼠血清代谢谱的变化,研究其代谢组学特征及其密切相关的代谢产物,探讨其分子机制及特征代谢物的潜在临床价值。方法灌胃30mg/kg百草枯建立百草枯中毒小鼠模型,应用GC-MS技术对血清样本进行检测,对检测结果应用主成分分析、正交偏最小二乘法-判别分析进行模式识别,然后通过变量重要性因子、t检验,结合数据库检索筛选出具有显著差异的代谢物。结果与对照组相比,模型组血清中的肌酐、脯氨酸、柠檬酸、琥珀酸、牛磺酸、苏糖酸、富马酸、甘氨酸、磷酸甲酯含量增加(P 0.05),而软脂酸、生育酚、色氨酸和6-磷酸葡糖酸含量减少(P 0.05)。结论通过对百草枯中毒模型小鼠血清代谢组学分析,证明代谢组学方法用于百草枯中毒研究是可行的,找到了与百草枯中毒密切相关的代谢信息,有助于阐明百草枯中毒机制。     

博落回中毒大鼠心肌组织代谢产物变化的研究

目的 基于气相色谱-质谱联用技术（GC-MS）的代谢组学方法研究博落回急性染毒模型大鼠心肌代谢产物的变化,寻找体内特征性代谢产物并探索相关毒性机制。方法 博落回总碱提取物溶液灌胃给药,剂量382 mg/kg,对照组给予相同剂量的空白溶液,利用GC-MS方法对心肌样本进行分析,利用偏最小二乘法-判别分析对结果进行模式识别,通过VIP(The variable importance in the projection)值> 1以及Student’s t检验（P <0.01）筛选出具有特征改变的差异代谢产物。结果 与对照组相比较,发现21种潜在的特征性代谢产物,经KEGG通路富集分析发现,这些代谢产物主要参与甘氨酸、丝氨酸和苏氨酸代谢途径、甘油酯代谢途径以及丙酮酸代谢途径。结论 经过对博落回染毒大鼠心肌代谢研究,发现了与中毒密切相关的代谢产物信息,为其中毒机制研究及法医学鉴定提供参考。     

丁卡因急性中毒致死小鼠血清和组织的代谢组学变化

目的采用代谢组学方法研究丁卡因急性中毒致死小鼠血清和组织(肾、肝和心脏)中的代谢物变化,寻找潜在的生物标志物及其相关代谢通路,为丁卡因急性中毒的死亡原因鉴定及毒理机制研究提供新思路。方法 40只ICR小鼠被随机分为对照组和丁卡因急性中毒致死组,以腹腔注射丁卡因建立急性中毒致死模型,运用超高效液相色谱-静电场轨道阱高分辨质谱联用(ultra-high performance liquid chroma-tography-electrostatic field orbitrap high resolution mass spectrometry,UPLC-Orbitrap HRMS)获取小鼠血清和组织的代谢轮廓。利用多元变量统计主成分分析和正交偏最小二乘-判别分析,并结合t检验和差异倍数分析找出与丁卡因急性中毒致死相关的差异代谢物。结果与对照组相比,丁卡因急性中毒致死组小鼠血清和组织的代谢轮廓表现出明显的区分。血清中鉴定出11种差异代谢物,包括黄嘌呤、精胺、3-羟基丁胺等;肝组织中鉴定出25种差异代谢物,包括腺苷酸、腺苷、柠檬酸等;心脏中鉴定出12种差异代谢物,包括次黄嘌呤、鸟嘌呤、鸟苷酸等;肾组织中鉴定出4种差异代谢物,包括牛磺鹅去氧胆酸、11,12-环氧二十碳三烯酸、二甲基乙醇胺和吲哚。丁卡因急性中毒主要影响了嘌呤代谢,三羧酸循环,丙氨酸、天冬氨酸和谷氨酸代谢。结论丁卡因急性中毒致死小鼠血清和组织中的差异代谢物有望成为该死因的候选生物标志物,该结果可为丁卡因急性中毒的作用机制和死亡原因鉴定提供研究基础。     

大鼠脑干早期创伤性轴索损伤的非靶向代谢组学研究

目的 通过非靶向代谢组学技术分析脑干创伤性轴索损伤后30 min的差异代谢物和关键代谢通路。方法将10只（8周龄,雄性）SD大鼠随机分为假手术组（Sham组）和脑干创伤性轴索损伤组（TAI组）,每组5只。采用Marmarou模型建立SD大鼠脑干创伤性轴索损伤模型,并以生命体征测量、HE染色和嗜银染色进行模型评价。采用UPLC/TripleTOF-MS系统分析TAI组和Sham组脑干组织的代谢物变化,筛选差异代谢物和分析差异代谢通路。结果 与Sham组相比,TAI组被打击后立即进入不同程度的昏迷状态,伴有生命体征紊乱,伤后30 min发现脑干存在弥漫性轴索损伤。代谢组学分析筛选出差异代谢物包括月桂酰二乙醇酰胺、邻苯二甲酸单乙基己基酯、PE(20:5(5Z,8Z,11Z,14Z,17Z)/24:1(15Z))、PC(16:0/16:1(9Z))、(2E,6Z)-十二二烯酸甲酯、六甲密胺,涉及代谢途径包括甘油磷脂代谢、甘油脂代谢、花生四烯酸代谢、逆行内源性大麻素信号、自噬-动物、糖基磷脂酰肌醇（GPI）-锚定生物合成。结论 利用UPLC/TripleTOF-MS技术进行大鼠脑干早期TAI的代...     

气相色谱质谱联用法检测大鼠尿液中2C-B及其代谢物

目的研究4-溴-2,5-二甲氧基苯乙胺（2C—B）在大鼠体内的代谢物以及代谢途径。方法取Wistar大鼠3只,以2C-B灌胃,收集24h内尿液,用B葡萄糖醛酸酶水解,Oasis HLB柱固相提取,DB-35MS柱分析,气相色谱质谱联用检测。结果从大鼠尿液中检出6种2C-B的代谢产物,分别为：4-溴-2-羟基-5-甲氧基苯乙醇、4-溴-2,5-二甲氧基苯乙醇、4-溴-2-羟基-5-甲氧基苯乙酸、4-溴-2,5-二甲氧基苯乙酸、1-乙酰氨基-2-（4-溴-5-羟基-2-甲氧基苯）乙烷和1-乙酰氨基-2-（4-溴-2-羟基-5-甲氧基苯）乙烷。未检出2C—B原药。结论2C-B在大鼠尿液中主要以代谢物形式存在,其在大鼠体内至少仔在两种代谢途径：第一种是2C—B的2位和5位氧上去甲基后氨基被乙酰化;第二种是2C—B去氨基生成醛,接着被还原或氧化生成醇和羧酸。     

液相色谱-串联质谱法检测生物样品中百草枯及其代谢物

目的建立生物样品中百草枯(paraquat,PQ)及其2种主要代谢物monoquat,paraquatmonopyridone(MP)的液相色谱-串联质谱(LC-MS/MS)检测方法,应用于百草枯中毒案件的法医学鉴定。方法生物样品经乙腈或甲醇沉淀蛋白,使用Agilent HILIC Plus(4.6×100mm,3.5μm)色谱柱,以0.1%甲酸水溶液～0.1%甲酸乙腈溶液(v/v)为流动相进行洗脱,在多反应监测模式下检测。结果百草枯及其代谢物在1～1000ng/mL内线性关系良好,相关系数(r)≥0.9996,最低检出限为0.34～6.00ng/mL,检测准确度为91.25%～113.44%,日内及日间精密度分别为1.51%～3.99%和1.92%～4.93%。结论本文建立的LC-MS/MS法具有灵敏度高、特异性好的特点,可应用于百草枯中毒相关案件的法医学鉴定。     

应用Pixel法测量平面内非直线长度的法医学研究

目的探寻一种法医鉴定工作中测量平面内非直线长度的方法。方法精确切断长度不同的10根焊接锡丝并读出结果作为对照组(钢尺直接读取法),再分别将这10根长度不同的焊接锡丝任意弯曲,然后用本文研究的方法计算出长度作为Pixel组,并对两组数据进行统计学分析。结果 Pixel方法得出的结果与钢尺读取法直接读出的结果之间的差异不具有统计学意义。结论 Pixel法在测量平面内非直线长度(如瘢痕、创口)上比传统的棉线法有明显优势,计算出来的长度更精确、易操作,使法医学鉴定结论更准确、有说服力,也更能适应诉讼要求,在法医学鉴定中有很好的应用前景。     

Are UK undergraduate Forensic Science degrees fit for purpose?

In October 2009 Skills for Justice published the social research paper ‘Fit for purpose?: Research into the provision of Forensic Science degree programmes in UK Higher Education Institutions.’ The research engaged employers representing 95% of UK Forensic Science providers and 79% of UK universities offering Forensic Science or Crime Scene degree programmes. In addition to this, the research collected the views of 430 students studying these degrees. In 2008 there were approximately 9000 people working in the Forensic Science sector in the UK. The research found that the numbers of students studying Forensic Science or Crime Scene degrees in the UK have more than doubled since 2002–03, from 2191 in to 5664 in 2007–08. Over the same period there were twice as many females as males studying for these degrees. The research concluded that Forensic Science degree programmes offered by UK universities were of a good quality and they provided the student with a positive learning experience but the content was not relevant for Forensic Science employers. This echoed similar research by the former Government Department for Innovation, Universities and Skills on graduates from wider science, technology, engineering and mathematics degree programmes. The research also found that 75% of students studying Forensic Science or Crime Scene degrees expected to have a career in the Forensic Science sector, meaning that ensuring these courses are relevant for employers is a key challenge for universities. This paper reflects on the original research and discusses the implications in light of recent government policy.     

道路交通事故伤残等级重新鉴定改变结论100例分析

目的研究道路交通事故伤残等级重新鉴定中暴露出来的初次鉴定中存在的问题,探讨解决方法。方法收集自2009年至2010年经上海市司法鉴定中心重新鉴定改变结论的道路交通事故伤残等级评定案例100例,对结论改变原因及其存在问题进行分析。结果鉴定时机把握不当、法医临床学检验方法不当、法医临床学检验不全面、医学影像学资料阅读错误、标准条款适用不当、肢体功能丧失程度计算错误、鉴定人鉴定理念就高不就低及其它一系列问题是重新鉴定改变结论的主要原因。结论针对上述问题应该从严格执业纪律,强化重新鉴定的实施,加强行政管理和业务培训等方面进行改进。     

The effect of NucleoSpin® Forensic Filters on DNA recovery from trace DNA swabs

Forensic DNA profiling is a globally accepted method for human identification, however, obtaining full DNA profiles from trace DNA can be challenging. The optimal recovery of DNA from trace DNA swabs is therefore crucial. Methods for extracting DNA from swabs often make use of a spin basket combined with a centrifugation step, to enhance the release of cells from the swab prior to DNA extraction. The NucleoSpin® Forensic Filter (Macherey-Nagel, Düren) is a type of spin basket, but it has not been thoroughly assessed on trace DNA samples. This study aimed to assess if the inclusion of the NucleoSpin® Forensic Filter significantly improved DNA recovery and DNA profiling success from cotton and flocked swabs used to collect trace DNA and buccal cells (control). Buccal cells and trace DNA samples were collected from 25 volunteers using each swab type (cotton and flocked) in duplicate. DNA was extracted from the samples using the NucleoSpin® DNA Forensic kit, one set with, and the other set without, NucleoSpin® Forensic Filters. DNA concentration was assessed using real time PCR, and DNA profiling was done using the PowerPlex® ESX 16 system. The inclusion of the NucleoSpin® Forensic Filters significantly improved DNA concentration for buccal cells that were collected using flocked swabs (p = 0.035). However, no significant differences were noted for trace DNA samples for either swab type. There was also no significant difference in DNA profiling success when NucleoSpin® Forensic Filters were used, regardless of swab and sample type. These results may be helpful for laboratories that are considering the NucleoSpin® Forensic Filters in the DNA extraction workflow, particularly for trace DNA samples.     

26例心脏传导系统发育异常猝死的病理学分析

目的研究心脏传导系统发育异常致猝死的病理变化。方法排除外伤、中毒及疾病所致死亡,收集26例不明原因猝死者心脏,采用作者创建的心脏传导系统取材法,常规切片、HE染色和Masson三色染色,光镜观察。结果 26例中见心传导系统有发育不全、结构异常、细胞或组织结构移位和窦房结纤维垫等病变,9例同时具有2~3种类型。16~30岁共21例,占全部样本的80.8%。结论心脏传导系统发育异常可能是不明原因青壮年猝死的重要原因之一。     

尿液中DNA的提取与检验

尿液中DNA的STR分型检验有一定的应用价值:国外文献报道过应用于运动员兴奋剂控制^[1]中可疑尿检样品的个体识别,国内则偶有案发现场提取到尿液、尿冰、尿斑并成功进行个体识别的案例报道。与血液、精斑、唾液等检材相比,尿液STR分型的检出率较低,主要是与尿液中的成份有关。本文重点分析人类尿液成分,分类总结目前可用于提取尿液中DNA的多种方法,分析可能影响尿液STR分型检验结果的因素。     

On the added value of forensic science and grand innovation challenges for the forensic community

In this paper the insights and results are presented of a long term and ongoing improvement effort within the Netherlands Forensic Institute (NFI) to establish a valuable innovation programme. From the overall perspective of the role and use of forensic science in the criminal justice system, the concepts of Forensic Information Value Added (FIVA) and Forensic Information Value Efficiency (FIVE) are introduced. From these concepts the key factors determining the added value of forensic investigations are discussed; Evidential Value, Relevance, Quality, Speed and Cost. By unravelling the added value of forensic science and combining this with the future needs and scientific and technological developments, six forensic grand challenges are introduced: i) Molecular Photo-fitting; ii) chemical imaging, profiling and age estimation of finger marks; iii) Advancing Forensic Medicine; iv) Objective Forensic Evaluation; v) the Digital Forensic Service Centre and vi) Real time In-Situ Chemical Identification. Finally, models for forensic innovation are presented that could lead to major international breakthroughs on all these six themes within a five year time span. This could cause a step change in the added value of forensic science and would make forensic investigative methods even more valuable than they already are today.     

基于Citespace的法医临床学研究发展历程分析

目的运用Citespace软件对法医临床学相关文献绘制知识图谱、进行可视化分析,探究该学科领域研究热点和动态发展,梳理鉴定难点脉络。方法汇总CNKI收录《中国法医学杂志》《刑事技术》《法医学杂志》《中国司法鉴定》刊登的法医临床学相关论文,利用Citespace分析其关键词、研究热点、基金资助、作者、发文机构及所在地等。结果法医临床学作为法医学分支学科之一,近年来呈现的科研成果部分获基金资助,多由开设法医学专业的院校完成;随着科研理论、技术方法推陈出新,法医临床学鉴定标准、版本也得以构建、更迭、完善,鉴定体量位居法医"四大类"之首。结论作为最具中国特色的法医学分支学科,法医临床学研究着重服务司法实践、以期用成果技术反哺鉴定,实现学术、实践的交流、转化。通过Citespace知识图谱可视化分析学科发展历程,有助于回溯此间规律,提示未来研究方向,提升鉴定业务水平,为持续增强法医临床学学科影响力提供借鉴。     

Civilian Forensic Technician and Sworn Police Officer Job-Related Stress

Forensic Technicians provide crime scene investigation services and are exposed to stressful violent crimes, motor vehicle accidents, biological or chemical hazards, and other appalling imagery. Forensic Technicians would likely experience physical and psychological stress after exposure to trauma, and security vulnerabilities similar to Sworn Police Officers. The perceived availability of mental health resources, job-related physical, psychological stress, and traumatic experiences of both Forensic Technicians and Sworn Police Officers from California law enforcement agencies were investigated using a self-reported survey. Responses were evaluated for any significant differences in the perceived stress, job-related physical stress, and resulting psychological impact affecting the participants. The survey contained a mix of True/False, Circle/Check the Appropriate Box, or Likert Scale (1–5) responses. The results were evaluated statistically and discussed. Results indicated Sworn Police Officers and Forensic Technicians have different on-duty stress levels, but similar off-duty stress levels. Nearly two-thirds of 54 job-related stressors were not significantly different between the two occupations. However, Forensic Technicians reported more adverse effects in 17 physical and psychological job-related activities compared with Sworn Police Officers. Forensic Technicians reported lower awareness levels and availability of agency mental health support services than were reported by Sworn Police Officers. This study reports for the first time an unexpected outcome that perceived and job-related psychological stress is greater for Forensic Technicians than Sworn Police Officers. Possible reasons for this disparity will be discussed as well as stress management tools that should be implemented to reduce health risk factors for both career professionals as well as increase public safety.