MALDI-TOF-IMS在蛋白质组学研究中的新进展

基质辅助激光解吸/电离-飞行时间质谱成像(matrix-assisted laser desorption/ionization time-of-flight imaging mass spectrometry,MALDI-TOF-IMS)技术已成为目前蛋白质组学研究中的经典技术,是一种研究生物组织切片中蛋白质和小分子物质分布的工具。MALDI-TOF-IMS通过单次扫描就能够同时分析生物组织切片中的各种未知组分,在保持组织中细胞和分子完整性的同时获取组织的分子成像图。近年来,质谱成像技术在生物医学各领域中有了较快发展,本文综合文献资料对MALDI-TOF-IMS技术目前的发展水平、质谱成像原理、方法学以及在法医病理学中的应用前景进行了综述。     

傅里叶显微红外光谱成像技术研究进展及其法医学应用

傅里叶显微红外光谱成像技术可收集微小物体的红外光谱信息,针对组织切片和细胞进行扫描,记录选择区域像素位点的红外光谱信息,并通过计算机软件构建出组织和细胞中光谱吸收峰的二维和三维光谱映射图像.由于红外光谱对生物大分子的敏感性,通过该技术可以有效分析组织和细胞的生化构成、分子分布、代谢变化.本文阐述了傅里叶显微红外光谱成像...     

质谱成像技术及其在法医毒理学中的应用

质谱成像(mass spectrometry imaging,MSI)是一种新型的成像技术,可在无标记的情况下同时检测并记录样品表面多种分子的空间分布信息。MSI的主要原理是将质谱与成像技术相结合,通过离子束或激光照射样品切片使其表面的分子离子化,经检测器获得质谱信号,再由成像软件将获得的数据转化成像素点并重构出目标化合物在组织表面的空间分布图像。MSI的样品制备包括样品收集与储存、组织切片、组织预处理、基质的选择与应用。目前,该技术已被广泛地应用于生物医学、新药研发以及蛋白质组学等领域,其在法医毒理学领域的应用也逐渐受到关注。本文综述了MSI的原理、样品制备过程,叙述了MSI在各种材料基质、草药混合物、潜指纹、毛发和动植物组织中的滥用物质及代谢物的应用,并对该技术在法医毒理学中的应用进行了展望,以期为MSI技术在法医毒理学中的应用提供思路和参考。     

干生物基质点微样技术及其在法医毒物学中的应用

干生物基质点(Dried Matrix Spot,DMS)微样技术是通过将微量生物液体(如血液、尿液和唾液等)收集于卡纸上并干燥后形成的样本采集和储存形式。干生物基质点微样技术较传统的采样方法有明显的优势:取样量小,样品处理简单,储存运输方便,目前已应用于流行病学调查、毒药物监测、药代动力学研究和毒代动力学研究等领域。通过对干生物基质的样品采集、储存稳定性、样品处理和分析方法等方面进行综述,以推动DMS微样技术在法医毒物学领域的拓展应用及鉴定技术的进步。     

MALDI-TOF-MS成像技术及其在法医损伤标志物筛选中的应用

传统的免疫学和分子生物学技术在寻找损伤分子标志物方面存在成果孤立、效价模糊以及相互间作用机制不明等缺点.质谱成像技术集筛选、分析和图像化于一体,更适合基于形态学的损伤分子标志物研究.本文综述了基于基质辅助激光解吸/电离-飞行时间质谱（matrix-assisted laser desorption/ionizationtime-of flight mass spectrometry,MALDI-TOF-MS）成像技术筛选并识别特征性蛋白质的方法和最新进展,阐述了其在法医学损伤研究中的应用价值.     

DNA甲基化在组织/体液来源鉴定中的研究进展

对可疑生物样本的组织/体液进行来源鉴别是重建犯罪现场、推断犯罪性质等侦查活动中极为重要的一环。对表观遗传学理论的研究证明运用基因组中存在的组织特异性DNA甲基化差异位点(t DMRs)可以对组织/体液进行来源鉴别。本文旨在通过对近年来DNA甲基化在法医学领域用于鉴定人体组织/体液来源方面的研究成果进行阐述,试图用所得到的信息来分析DNA甲基化作为一种组织/体液鉴定遗传学标记的可能性、优劣点及其应用价值和发展前景,以期能为法医工作者的相关研究及实践提供参考。     

中毒致死者体内芬氟拉明的GC/NPD,GC/MS法测定

建立了生物检材中芬氟拉明的定性定量分析方法。体液及脏器组织经有机溶剂提取后 ,用GC/MS法进行药物筛选、定性 ,生物检材中的芬氟拉明浓度用4 -苯丁胺作内标、GC/NPD法测定。测得芬氟拉明中毒致死者的血液、尿液、肝等组织中浓度分别为7.8μg/ml、64.2μg/ml、31.3μg/g。并对尸体解剖所见及方法可行性进行讨论     

指(趾)甲在滥用药物分析中的研究现状及应用展望

在法医毒物分析中,各类生物检材都具有其特点和应用范围。本文对指(趾)甲的生物学特征、样品的采集和前处理、指甲中部分毒(药)物的分析方法作了较为详细的综述,介绍了指(趾)甲中滥用药物含量的影响因素,并针对研究中的问题,结合已有研究对其应用前景作出展望,认为指(趾)甲作为一种在体内滥用药物分析中尚未被普遍应用的非常规生物检材,在法医毒物学方面拥有潜在的可用性和优势。     

HE染色切片组织的STR检测及法医学应用

目的建立并优化HE染色切片组织STR分型方法,评估HE染色切片组织的STR分型有效性。方法用QIAgen法提取2例尸检人体的心、肝、肺、肠等8种组织的HE染色切片DNA,用TaqMan荧光探针法通过荧光域值循环数(Ct值)测定获得各提取液中的DNA质量浓度,以阳性内对照(internal posi-tive control,IPC)监测PCR过程中的抑制水平。再用Identifiler试剂盒扩增,在3100-Avant上进行STR片段分析。结果在本研究建立优化的STR分型技术条件下,8种人体组织的HE染色切片DNA提取液质量浓度均可达到1ng/μL以上,其IPC的Ct值提示无PCR抑制剂。HE染色切片组织的STR分型有效性与石蜡包埋组织相当,其DNA随时间延续而缓慢降解。结论在一定保存时限内,HE染色切片的DNA质和量可以满足STR分型检测需要,但受残余甲醛固定剂的影响,HE染色切片的分型成功率随保存时间延长而降低。     

傅里叶红外光谱技术在法医学脑挫裂伤鉴定中的应用展望

脑挫裂伤是常见的一类颅脑损伤,单纯的组织病理学技术可能难以早期发现及精确诊断。傅里叶变换红外面扫描成像技术(FTIR-mapping)是病理学研究领域的新技术,因其结合了傅里叶变换红外光谱、红外显微镜技术以及面扫描成像技术的特点,具有测定方法简便、对样本无损、灵敏度高、准确、直观等优点,突破了传统技术的局限,能检测出发生了物质数量、结构以及构象变化但缺乏明显形态学变化的脑挫裂伤,从而实现早期精准地诊断脑挫裂伤。FTIR-mapping虽已能绘制某些损伤组织的红外光谱病理图像,但其在法医病理学中的价值有待全面深入开发。本文综述了脑挫裂伤的法医病理学研究进展以及傅里叶变换红外面扫描成像技术在法医病理学中的应用。     

International Conference on Harmonisation; guidance on Q1A stability testing of new drug substances and products; availability. Notice

The Food and Drug Administration (FDA) is announcing the availability of a revised guidance entitled "Q1A(R) Stability Testing of New Drug Substances and Products." The revised guidance was prepared under the auspices of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). The guidance sets forth recommendations on the information to be submitted in the stability data package for a new drug substance or drug product for a registration application within the three regions of the European Union (EU), Japan, and the United States. The purpose of there vision is to add information to certain sections and to provide clarification to other sections of the guidance.     

利用额窦影像进行个体识别的研究进展

鉴于额窦不规则气腔形态的高度特异性、成熟后形态不变的稳定性以及放射学技术在临床的广泛应用,额窦影像已成为法医学个体识别的重要研究对象。利用额窦影像进行个体识别已具有法庭意义。当DNA和指纹等传统个体识别方法应用受限或无法有效进行,或缺乏相应的牙科记录等情况时,额窦影像对比是有效的个体识别替代方法。多种影像资料都可用于额窦的个体识别,以人工视觉观测分析为主要方式,受专业性要求高、效率低及应用范围小等限制,无法有效应用于大型灾难等情况。近年来,一些计算机图像识别技术被应用于额窦影像的识别中,能显著提高识别的效率,有望克服人工识别效率低等应用困难。本研究对额窦影像用于法医学个体识别的报道进行总结,以回顾额窦影像用于个体识别的研究进展,为额窦影像的进一步研究提供参考,为探索及建立一套更加快捷、高效及准确的个体识别方法提供思路。     

Immunofluorescence detection of drugs in postmortem tissues: a new technique with potential for assessment of drug influence in cause of death

This report describes a new technique, immunofluorescence, for the detection and possible characterization of drug content in postmortem tissues. By using antisera generated against a drug-protein conjugate, the stabilization of tissue-sequestered drug is accomplished by incubation of fresh frozen sections of tissue with dilute solutions of rabbit anti-drug antibodies. Secondary incubation with a fluorescence-labeled anti-rabbit immunoglobulin labels these points of sequestration. Tissue sections so stained are examined by fluorescence microscopy. In studies with rats given graded doses of morphine sulfate, there were discernible differences in tissue binding of morphine in brain sections from animals treated "therapeutically," fatally, and chronically. Extension of these studies to human autopsy material is anticipated and potential problems are discussed. This technique offers the forensic toxicologist the potential for evaluating the drug content of tissues in situ.     

Morphometric assessment of testicular changes in drug-related fatalities

Testicular sections of 50 deceased drug addicts were examined by light microscopy using morphometric methods and determining the extent of spermatogenesis by application of the Johnsen score count. Twenty cases of violent death served as reference sample. The study was aimed at obtaining data about the degree of germinal gland damage following the duration of the individual drug career, which was correlated to the histologic findings. Progressively reduced maturation of germinal cells, decrease of tubular diameter and height of sperm-producing epithelium were found the longer the individual case history has been. The results clearly indicated a substantial impairment of male fertility in the course of a drug career, but etiology seems to be multifactorial, since most of the deceased were polytoxicomanic and more than 40% were under the influence of alcohol at the time of death. Testicular damage is obviously a sequel of long-lasting abuse and a very complex process, presumably progressing with the duration of the adverse habit.     

Progress in diagnosis of neurogenic erectile dysfunction

Recently, with application of evoked potentials technology in the test of somatic and autonomic nerves, quantitative sensory testing in the detection of small nerve fiber function, and functional magnetic resonance imaging in the detection of senior central function, the detection of neural function has become more accurate. This article reviews the progress and application of diagnostic methods about neurogenic erectile dysfunction in order to provide a reference for forensic diagnosis and research in the future.     

Lymph-node and thymus pathology in fatal drug addiction

Lymph-node sections from the porta hepatis, the lung hilus, the para-aortic and axillary regions and from the neck from totally 50 drug addicts submitted for medico-legal autopsy at the Institute of Forensic Medicine in Copenhagen were studied together with tissue sections from 23 normal persons. In addition thymus sections from 30 drug addicts and 20 normal persons were investigated. Enlargement of lymph-nodes was found significantly more often in active drug addicts compared to normals except for the nodes on the neck. Birefringent material was seen in portal lymph-nodes in 42% of addicts, in para-aortic and lung hilus nodes in 18% each and in the axillary nodes in 12%. Signs of antigenstimulation evaluated by the number of germinal centre and plasma cells were found in about 60% or more in active drug addicts compared to about 30-40% in normals. There was not significant relation between the size of the lymph-nodes and the immunoactivity nor to the duration of the abuse. Examination of the thymus showed that the average weight in active drug addicts was 34 g, in normals 25 g. Histologically the tissue was in 48% of the active addicts composed of more than 80% lymphatic tissue compared to 15% of the normals. The results indicate that the enlargement and histological signs of activation in all the lymph-nodes investigated are caused by continuous antigenstimulation due to repeated injections of various antigens. The same may in part be applied to the thymus changes demonstrated.     

CT技术在法医人类学研究中应用的进展

近年来,将计算机断层扫描(CT)等影像学新技术应用于法医人类学个体识别的研究受到国内外法医人类学家的重视。CT具有清晰快速的成像能力以及强大的后处理功能,无需骨骼前处理就能提供高分辨率模型图像,对法医人类学具有重要的研究和应用价值。本文就CT技术在法医人类学研究中应用的进展进行综述,以期为相关研究和实践提供参考和借鉴。     

视野空间注意的事件相关电位研究进展

在视野脑机制的研究中,事件相关电位（event-related potential,ERP）具有高时间分辨率的特点。近年来高密度记录技术和脑成像技术的发展,提高了ERP在脑功能定位方面的应用。本文概述了近年来有关ERP在视野空间注意研究的实验方法、结果和初步结论,并提出今后相关问题的研究方向及法医学意义。     

Spleen and portal lymphnode pathology in fatal drug addiction

Spleen and portal lymphnode sections from 86 drug addicts submitted for medico-legal autopsy at the Institute of Forensic Medicine in Copenhagen in the year 1979 were studied together with tissue sections from 24 "normal" persons. In 70% of the drug addicts the spleen weight was more than 200 g, and in 71% portal lymphnode hyperplasia was found. Birefringent foreign material was found in spleen tissue of drug addicts in 72% and in portal lymphnode tissue in 44%. Signs of antigen stimulation in both spleen and portal lymphnode tissue evaluated by the number of germinal centre and plasma cells were found in more than 80% of the drug addicts compared with about 20% of the "normal" persons. The results were related to anamnestic information of duration of drug abuse, to the spleen weight, to the occurrence of birefringent material and to the liver changes. Examination of lysozyme and immunoglobulin containing cells using the indirect preoxidase technique was performed in a total of 72 cases of spleen tissue, 59 cases of portal lymphnode tissue from drug addicts, 24 cases of spleen tissue and 18 of portal lymphnode tissue from "normal" persons. Lysozyme, IgM and IgG containing cells were found significantly more often among drug addicts than "normal" persons. The results indicate that the splenomegalia and the portal lymphnode hyperplasia often found in drug addicts are caused by continuous antigen stimulation due to repeated injections of various antigens.     

Body packing: the value of modern imaging procedures in the detection of intracorporeal transport media

The necessity of rapid objective appraisal of the suspicion of intracorporeal drug smuggling (body-packing) by effective methods of investigation in the course of the first criminal investigation department measures raises the questions of suitable and admissible methods. The proportion of undetected crimes in intracorporeal narcotic smuggling must be rated as very high according to the data of the Federal Criminal Investigation Department. In the present paper, the suitability of various imaging techniques for drug detection are reported in terms of their risk, practicability and costs. For this purpose, the value of digital radiography, two-spectra radiography and X-ray computer tomography as compared to conventional X-ray investigations is examined in human experiments. A reality-oriented narcotics dummy (glucose pressed hard in preservative) was developed and administered to nine volunteers per os with a variable initial alimentary situation. Four radiograms were taken at fixed times up to 12 hours after administration with each imaging technique. The highest rate of detection was attained with computer tomography. In contrast to the other methods, more than 90% of the body packs could be identified here. Nevertheless, an application in criminal investigation practice cannot be recommended owing to various disadvantages. The recovery rate of the remaining methods is between 20% and 25%.