ERCC1和XPF基因在法医学年龄推断中的初步研究

目的探讨ERCC1和XPF基因mRNA及蛋白在不同年龄段健康汉族人群中的表达情况,分析其mRNA和蛋白表达量与年龄之间的相关性,以期为法医学年龄推断提供新的分子生物学指标。方法收集150例不同年龄段健康汉族人群外周血样,梯度离心法分离血浆,Trizol法提取外周血单个核细胞(PBMCs)总RNA;实时荧光定量PCR检测ERCC1和XPF基因mRNA在PBMCs的相对表达量;酶联免疫吸附试验检测ERCC1和XPF蛋白在血浆中的表达量。结果 ERCC1和XPF基因mRNA在不同性别PBMCs的相对表达量均无统计差异(P0.05)。ERCC1和XPF基因mRNA相对表达量在不同年龄段有统计学差异(P0.05),且不同年龄段组间比较亦均有统计学差异(P0.05)。回归分析显示ERCC1和XPF基因mRNA相对表达量与年龄呈负相关,其相关系数(r)分别为-0.578和-0.844;以年龄为自变量(x),以mRNA相对表达量为因变量(Y),其拟合曲线分别为Y=3.3E-5x~2-0.0261x+1.9175(R~2=0.3244,P0.01)、Y=0.0003x~2-0.0459x+2.0439(R~2=0.729,P0.01)。血浆中ERCC1和XPF蛋白表达量在不同年龄段及性别间均无统计差异(P0.05)。结论 ERCC1和XPF基因mRNA在PBMCs的相对表达量随年龄增加而下降,其血浆中蛋白表达与年龄无关,为建立ERCC1和XPF基因与年龄之间的数学模型提供理论学依据。     

人外周血FoxO3a mRNA表达随龄变化规律的初步研究

目的评估人外周血中FoxO3a mRNA表达的随龄变化规律,为法医学个体年龄推断寻找新的分子指标。方法采集80个健康无关个体血样,按年龄分为8组:≤9岁、10～19岁、20～29岁、30～39岁、40～49岁、50～59岁、60～69岁、≥70岁组,每组10人。提取总RNA后,用实时定量RT-PCR技术检测FoxO3a mRNA的表达水平。结果人外周血FoxO3a mRNA表达随年龄的增长而逐渐增加,具有良好的相关性(r=0.841,P0.01),以年龄分组为自变量,FoxO3a mRNA表达水平为因变量,可得拟合曲线Y=0.001e0.418x。结论人外周血中FoxO3a mRNA表达具有较强的年龄特征,有望成为年龄推断的有用指标之一。     

大鼠骨骼肌挫伤后Fzd2表达与损伤时间的关系

目的检测大鼠骨骼肌挫伤后卷曲受体蛋白2(frizzled-2,Fzd2)mRNA及其蛋白表达量与损伤时间的关系,探讨其是否可以作为损伤时间推断的指标。方法利用RT-qPCR和Western印迹法检测正常对照组及损伤后4~48 h每4 h大鼠骨骼肌中Fzd2 mRNA和蛋白水平的表达量。结果 Fzd2 mRNA在损伤后24 h、36 h、40 h相对表达量增高,24 h组表达量达正常对照组的两倍(P0.05);而Fzd2蛋白在损伤后相对表达量变化不明显(P0.05)。结论大鼠骨骼肌损伤后Fzd2 mRNA在一定时间内的表达变化情况可以作为多指标联合推断损伤时间的依据。     

大鼠早期缺血心肌中Basigin mRNA的表达

目的检测Basigin mRNA在大鼠早期缺血心肌和非缺血区心肌中的表达差异及探讨其法医学意义。方法建立大鼠早期心肌缺血手术模型,分为EIM组、NIM组、假手术组以及空白对照组,采用real-time PCR方法检测大鼠心肌缺血后15min、30min、1h和2h Basigin mRNA的表达量。结果与NIM组、假手术组及空白组相比,EIM组BSG mRNA表达量在缺血15min时降低,而冠状动脉结扎1h后表达量降低到SO组的一半,此时具有统计学差异(P0.05),结扎2h后表达量恢复至SO组水平。NIM组、SO组和空白对照组之间表达量无统计学差异(P0.05)。结论心肌急性缺血2小时内BSG mRNA表达降低明显,之后升高,说明BSG参与早期缺血心肌自我保护病理生理过程,提示BSG可用于鉴定早期心肌缺血。     

大鼠脑损伤后EPO及其受体表达与损伤时间的关系

目的研究大鼠脑损伤后脑组织中促红细胞生成素(erythropoietin,EPO)及其受体(erythropoietin receptor,EPOR)的表达与损伤时间的关系。方法 72只SD大鼠随机分为对照组(36只)、脑损伤组(36只),在建模后1、2、4、8、12、24 h每组各取6只大鼠处死,取挫伤部位脑组织,应用实时荧光定量PCR和Western印迹法检测不同时间点EPO以及EPOR的mRNA表达和蛋白表达情况。结果伤后24h内,EPO及EPOR的表达随损伤时间的增加而呈上升趋势。EPO的mRNA及蛋白表达与损伤时间之间存在相关性,其相关系数分别为0.875、0.911(P0.05);EPOR的mRNA及蛋白表达与损伤时间之间存在相关性,其相关系数分别为0.936、0.905(P0.05)。结论大鼠颅脑损伤早期EPO及EPOR的表达呈逐步上升趋势,与损伤时间具有相关性,可以作为脑损伤时间推断的参考。     

Spry1和MAPK蛋白在病毒性心肌炎心肌组织中的表达

目的通过探讨Spry1和MAPK蛋白在病毒性心肌炎(viral myocarditis,VMC)心肌组织中的表达,揭示其发病及猝死机制,并为法医学心源性猝死的鉴定提供指导。方法 30只Balb/c雄性小鼠随机分为VMC组和对照组,分别腹腔注射柯萨奇B3病毒和Eagel’s培养液,处死小鼠取心脏,进行常规病理检查,并通过免疫组织化学、Western印迹法及real-time PCR方法检测心肌组织中Spry1蛋白及mRNA、MAPK蛋白表达变化。结果 VMC组在光镜下可见心肌间质水肿,广泛炎症细胞浸润,心肌细胞坏死,心肌纤维形成多处灶性、大片状坏死,心肌组织中Spry1蛋白表达水平低于对照组(P0.05),Spry1 mRNA水平略降低(P0.05);而MAPK蛋白表达水平在VMC组高于对照组(P0.05)。结论 Spry1参与的MAPK/ERK通路在病毒性心肌炎心肌纤维化过程中起着促进胶原表达的作用,从而导致心律失常、心功能减退甚至心源性猝死。     

Cathepsin-L在缺血心肌中的表达

目的检测cathepsin-L(CTSL)在大鼠早期缺血心肌中的表达变化,并探讨其法医学意义。方法建立大鼠早期心肌缺血模型,设置早期缺血组(early ischemic myocardium,EIM)、非缺血组(non-ischemic myocardium,NIM)、假手术组以及空白对照组,采用real-time PCR方法检测大鼠缺血后15min,30min,1h和2h CTSL mRNA的表达量,并进行组内和组间比较。收集心源性猝死者缺血心肌和机械性损伤致死者正常心肌样本,采用免疫组化染色方法观察心肌组织中CTSL蛋白表达。结果 EIM组CTSL mRNA表达量与NIM组、假手术组及空白组相比,在15min时无统计学意义(P>0.05),在30min、1h和2h时分别升高1.4、3.1和4.5倍(P<0.05);其余3组间差异无统计学意义(P>0.05)。观察10例冠脉狭窄程度Ⅲ～Ⅳ级的心源性猝死(SCD)者缺血心肌,CTSL表达增强。结论大鼠心肌缺血后早期即可检测到CTSL mRNA的升高,并在SCD冠脉狭窄死者缺血心肌中高表达,提示CTSL可作为心肌缺血与SCD的参考指标。     

创伤性脑损伤大鼠水通道蛋白4表达变化及法医学意义

目的观察大鼠颅脑损伤后不同时间内水通道蛋白4（AQP4）mRNA表达的变化，探讨AQP4在脑损伤经过时间推断中的意义。方法利用液压冲击法制作不同程度大鼠颅脑损伤模型，在伤后不同时间（0．5、2、6、12、24、48、72h），应用RT-PCR法检测脑组织AQP4 mRNA表达，同时以非损伤组做对照。结果不同程度颅脑损伤后0．5h脑组织AQP4 mRNA表达均开始上调（P〈0．01），6、12h依次增高，24h达到高峰，差异均有统计学意义（P〈0．01），72h时仍维持较高水平（P〈0．01）；0．5h时轻度、中度、重度脑损伤时AQP4 mRNA的表达差异两两之间无统计学意义（P〉0．05），2、6、12、24、48、72h轻度、中度、重度脑损伤时AQP4 mRNA的表达差异两两之间有统计学意义（P〈0．01）。结论颅脑损伤后AQP4 mRNA呈现出时序性变化，其变化规律可望成为法医学推断早期脑损伤时间的指标之一。     

河南与浙江地区汉族女性青少年骨发育差异性分析

目的 探讨河南与浙江地区之间女性青少年骨发育的差异性.方法 摄取河南及浙江两地599例年龄介于12～19岁的女性锁骨胸骨端及六大关节x线片.对照青少年骨发育分级标准24项骨骼发育指标进行阅片,并通过数学模型法计算骨龄,然后利用SPSS 15.0软件对骨龄与生活年龄的比值进行不同地区间差异性分析. 结果 河南女性青少年骨发育在14-15岁年龄段(不含15周岁)比浙江提前约0.39岁,河南女性青少年骨发育在15～16岁年龄段(不含16周岁)比浙江滞后约0.37岁,河南女性青少年骨发育在18-19岁年龄段(不含19周岁)比浙江滞后约0.38岁,余年龄段无统计学差异.结论 河南与浙江地区女性青少年骨发育比较存在差异性,呈现先快后慢的趋势.本研究结论 可供法医学、人类学、临床医学等有关学科对青少年骨龄评价作为参考.     

大鼠局灶性脑挫伤后Cdk5的表达变化

目的研究大鼠受到一定钝力打击造成中度脑损伤后,3h至10d脑组织Cdk5的表达规律及法医学意义。方法建立大鼠钝力性局灶性脑挫伤模型,应用免疫组织化学技术和免疫蛋白印迹技术观察脑损伤后不同时间脑组织中Cdk5表达变化。结果正常组、假手术组脑组织有少量Cdk5表达;实验组损伤后3h组、6h组Cdk5表达增高,12h组表达明显升高(P0.05);在24h达到峰值,3d组、5d组、7d组、10d组逐渐下降,并在7d组、10d组基本降至正常,与正常组、假手术组比较基本无差异性(P0.05)。结论大鼠钝力性脑挫伤后Cdk5在挫伤脑组织周边区表达量增多,并呈现单峰变化,随时间延长基本降至正常。Cdk5表达变化可为早期脑挫伤时间推断奠定基础,提供新的参考指标。     

Investigation of homicides interred in concrete--the Los Angeles experience

Decedents interred in concrete present unique problems and investigation of these deaths necessitates a team of forensic specialists. The Los Angeles County Department of Coroner has had five such deaths in the past 18 years. The buried cases needed layer-by-layer excavation to establish the time and cause of death. Metal detectors are often used in this process. X-rays of the interred remains were completed to help with locating the decedent's position in the concrete. The breaking of concrete in some of the cases required the use of a sledgehammer and later a chisel in a manner that would not damage the remains. Postmortem dismemberment was frequent in our cases. The decedents were all female or prepubescent children, and the perpetrators were closely related to the decedents. While concrete can interfere with determination of postmortem interval, it can also preserve the remains and assist with identification.     

Forensic Approach to an Archaeological Casework of “Vampire” Skeletal Remains in Venice: Odontological and Anthropological Prospectus*

Abstract: During the years 2006–2007, the Archeological Superintendent of Veneto (Italy) promoted a research project on mass graves located on Nuovo Lazzaretto in Venice, where the corpses of plague deaths were buried during the 16th and 17th centuries. The burials were of different stages and are believed to be the remains of plague victims from the numerous outbreaks of pestilence, which occurred between the 15th and 17th centuries. Among the fragmented and commingled human bones, an unusual burial was found. The body was laid supine, with the top half of the thorax intact, arms parallel to the rachis axis, the articulations were anatomically unaltered. Both the skull morphology and the dimensions of the caput omeris suggest the body was a woman. A brick of moderate size was found inside the oral cavity, keeping the mandible wide open. The data collected by the anthropologist were used to generate a taphonomic profile, which precluded the positioning of the brick being accidental. Likewise, the probability of the brick having come from the surrounding burial sediment was rejected, as the only other inclusions found were bone fragments from previous burials in the same area. The data collected by the odontologist were employed for age estimation and radiological dental assessment. The forensic profile was based conceptually on the “circumstances of death” and concluded that the positioning of the brick was intentional, and attributed to a symbolic burial ritual. This ritual confirms the intimate belief held at those times, between the plague and the mythological character of the vampire.     

Forensic Analysis of a Contact Lens in a Murder Case

Contact lenses have had rare relevance in trials and/or investigations. After 5 years of burial, orbital remnants were retrieved from an exhumed body and subsequently identified as a key piece of material evidence in a murder trial. The exhumed case materials were evaluated under laboratory conditions and were determined to be contact lens remnants. Contact lens fracture and burial simulation studies were performed to provide additional corroboration of the physical findings of the exhumed contact lens remnants. This material evidence was instrumental in providing factual proof refuting the defendant's testimony in the murder trial. A brief history of contact lens composition and use is provided for understanding the methods and observational results. This forensic case study represents the first published documentation of a contact lens from an exhumed body being used in a murder investigation and establishes an operational procedure for future forensic contact lens examinations.     

毒物代谢动力学研究在法庭科学中的应用

随着法庭科学的发展以及近年来涉及毒品、毒物等中毒案件的增加,许多研究者对毒物代谢动力学进行研究,并取得了一定的成果.本文综述毒物代谢动力学研究及其在法庭科学中的应用状况,概要介绍毒物代谢动力学的研究现状、研究方法及研究成果,以期为相关研究及实践提供参考和借鉴.     

Antemortem and Postmortem Nonapposite Data—A Multidisciplinary Identification Strategy

Identification of human remains is often achieved by comparing documented reference data with the same type of evidence obtained from the remains. We present a case of a decomposed unidentified body, whose identity was presumed but because of the low validity range of the available data, the identification process could not be completed. Antemortem radiographs of the teeth found in the house of the reputed victim could not be compared to the edentulous cadaver and the kinship between the victim and the only living relative that could provide DNA for comparison was too tenuous. Isolated teeth found at the scene, were neither a source of DNA reference information nor questioned data to be compared to the antemortem radiographs. The strategy implemented by the investigators to reallocate the status of the isolated teeth from the questioned source to the reference source of DNA to be compared with the cadaver is presented.     

Comparison Among Manual Facial Approximations Conducted by Two Methodological Approaches of Face Prediction

This study verified the difference between two methods of forensic facial approximation (FFA) regarding recognition and resemblance rates. Three‐dimensional models of skulls were obtained from computerized tomography (CT) scans of two subjects (targets). Two manual FFAs were performed for each target, by applying two different guidelines for the facial structures (what we called “American method” (AM) and “Combined method” (CM)). Unfamiliar assessors evaluated the sculptures by recognition and resemblance tests. The AM was that which allowed more correct responses of recognition and higher resemblance's scores for the male target (p < 0.001). Regarding guidelines for modeling characteristics of the face, the ones that are practical and easily performed for sculptures, such as the length of the anterior nasal spine multiplied by 3 for nose prediction, may offer better results in terms of resemblance.     

Creating a State‐Academic Partnership to Advance a Forensic Teaching Service: Benefits and Barriers

In Washington State, like many states, there is a shortage of forensically trained mental health clinicians to work with criminal justice‐involved individuals. At the direction of the state legislature, a collaborative project was undertaken by the University of Washington, the state Department of Social and Health Services, and a state psychiatric hospital to develop a proposal for a jointly sponsored forensic teaching service. The authors reviewed the literature, surveyed and interviewed forensic psychiatry and psychology training directors, and conducted site visits of selected training programs that offer multidisciplinary training or have affiliations with state hospitals. The authors conducted focus groups of additional stakeholders, including clinicians and patients in forensic settings, to better understand the needs in Washington. The authors report on several common benefits and barriers to establishing forensic teaching services. Other states and forensic programs may find this article useful in identifying common considerations for forensic mental health teaching services.     

刑事强制鉴定制度初探

近年来,随着刑事要案为公众所广泛关注,在某些案件中,司法鉴定成为直接影响被告人刑事责任的重要手段,也被推至舆论的风口浪尖之上。然而学界却少有对强制鉴定构建模式的专门论证。我国应当在法律中规定当事人的鉴定请求权,构建与裁量鉴定并行的强制鉴定制度,确定其适用条件、具体程序及监督体制。     

Pacemaker/defibrillator evaluation at Los Angeles County Department of Coroner*

Pacemakers and implantable cardioverter-defibrillators (ICDs) are implanted medical devices for the treatment of cardiac arrhythmias. These devices are now commonly encountered in the postmortem situation. The Los Angeles County Department of the Coroner uses the services of a cardiac electrophysiology consultant to interrogate pacemakers and ICDs, producing a detailed picture of the cardiac events recorded by the device. We have used this method to evaluate 20 cases where the ICD or pacemaker provided significant information. We report four forensic cases to illustrate the different applications of pacemaker interrogation and its contribution in forensic investigation. This technique was used to establish identification of the decedent, to determine the cause and time of death, and to determine whether device malfunction could have played a role in the death. We conclude that detailed evaluation of the pacemaker or ICD in selected cases may provide essential information to the forensic pathologist.     

道路交通事故法医检验技术的研究进展

近年来,经济快速发展和机动车数量急剧增多,道路交通事故频发和伤亡人数不断上升已经成为备受关注的全球性问题。法医检验作为交通事故处理中重要的证据,其鉴定意见直接影响着当事方的权益和事故处理。不容置疑,法医检验技术的运用对鉴定意见有十分重要的影响作用。本文对国内外法医检验技术的最新研究进展和应用进行综述,供同行实践工作中参考。