月经血鉴定与基质金属蛋白酶-11

月经血鉴定是法医血痕检验的内容之一。本文就月经血鉴定的几种经典方法进行回顾,并分析其应用局限性。基质金属蛋白酶-11是一种能降解细胞外基质的水解蛋白酶,有研究提示基质金属蛋白酶-11以其在月经血中表达的特异性和检测方法的灵敏性有可能成为对月经血鉴定的新一代标志物。本文对基质金属蛋白酶-11的结构、功能、在月经血鉴定中的应用及展望进行综述。     

免疫组织化学检测MMP-11鉴定月经血

目的建立用自制兔抗人基质金属蛋白酶-11（matrix metalloproteinase-11,MMP-11）多克隆抗体鉴定月经血的免疫组化方法,探讨MMP-11蛋白酶的细胞定位。方法运用链霉卵白素-碱性磷酸酶（SAP）免疫组化法检测人月经血及外周静脉血涂片、阴道液涂片、子宫内膜石蜡切片和陈旧月经血痕涂片中的MMP-11。结果MMP-11存在于子宫内膜上皮细胞和间质细胞内;外周静脉血和阴道液涂片未见有MMP-11染色;陈旧月经血痕涂片罕见完整子宫内膜细胞,但能检测到微弱特异信号。结论用自制抗MMP-11多克隆抗体采用免疫组化技术能有效地检测MMP-11,区分月经血和静脉血。     

D-dimer assays for the identification of menstrual blood

A method to reliably distinguish menstrual blood from blood in the normal circulation (peripheral blood) would be of considerable use in the forensic analysis and interpretation of evidence in sexual offence investigations. Previous attempts to address this issue have explored microscopy, lactate dehydrogenase isozyme identification, mRNA and miRNA profiling, and identification of the products of fibrinolysis. Here, four assays for D-dimer, a terminal degradation product of fibrinolysis, are evaluated for their specificity and sensitivity in detection of menstrual blood. In addition the effect of exercise, and sample storage upon D-dimer detection was investigated. Comparison of different assays revealed significant differences in results given. Nevertheless, no positive results for D-dimer were obtained using peripheral blood, mixtures of peripheral blood with semen, or peripheral blood taken from donors after moderate exercise. D-dimer was found to be detectable in 100% of menstrual blood samples after 1 week at room temperature and also in samples stored long-term (>3 years) at -20 °C. D-dimer may be an effective, simple to use tool for the presumptive identification of menstrual blood identification.     

Detection of blood opiates by fluorescence-polarization immunoassay

It was found possible to use fluorescence-polarization immunoassay (FPIA) in screening of opium drugs in blood and blood serum as a preliminary test. The results of the study of blood and serum samples by FPIA and chromato-mass-spectroscopy were compared. As a criterion of assessment of the preliminary study results we propose to use a cutoff point by serial production of the analysed blood samples of the threshold morphine concentrations (20 ng/ml for living subjects and 40 ng/ml--for corpses). The method of opiates screening in blood proposed is characterized by fast conduction, high specificity and precision, small amounts of the expert material.     

Development of an immunoassay for the differentiation of menstrual blood from peripheral blood

This study examined all unintentional firearm fatalities while hunting that occurred in Sweden between 1983 through 2008. The circumstances as well as the impact of the hunter's exam on fatality frequency were analysed. During these 26 years, there were 48 such fatalities, representing 53% of all (n=90) unintentional firearm deaths during the same period. The average annual number of fatalities decreased over the last few decades. Very restrictive firearm legislation in Sweden combined with the introduction of a mandatory hunter's exam since 1985 accounted, at least partly, for this finding. Moose hunting accounted for 46% of the fatalities and small game hunting for the remaining cases. The mean age of the victims was 50 years and 96% of them were males; all shooters were males. During moose hunting, most of the victims were mistaken for game, whereas in small game hunting most of the fatalities were related to falls and improper handling of the weapon. Human error was thus the main cause of these fatalities.     

Detection of cocaine metabolite in perspiration stain, menstrual bloodstain, and hair

Low nanogram and picogram quantities of cocaine metabolite equivalents were detected in extracts from perspiration stains, menstrual bloodstains, and hair using radioimmunoassay. The theory of drug inclusion in hair and its significance are discussed.     

Detection of cocaine in blood stains]

Cocaine is rapidly degraded in blood samples, and its degradation was found to be highly dynamic in nature. The analysis of blood spots dried on filter paper may provide a method to minimize the break-down of cocaine and to largely preserve the analytical profile of the parent drug and its hydrolysis products at the time of sampling. The short term stability of cocaine in 100 microL blood spots prepared from unpreserved and preserved (sodium fluoride, 0.25%) blood samples was compared to the stability of the particular whole blood specimens stored in tubes at ambient temperature and at -20 degrees C. Due to dehydration, both the chemical and the enzymatic hydrolysis of cocaine and its products could be stopped in dried blood spots. More than 75% of the initial cocaine concentration could be detected in the blood spots, and the analytical profile was ensured for 17 days. Provided its practical suitability, the spot technology should offer a simple approach to detect actual impairment of motorists taken in police custody in the view of section 24a of the German traffic act as well as in cocaine associated criminal cases.     

Detection of blood in stains on material evidence by fluorescent hemotest and spectrofluorometry

The efficiency of detection of blood in stains on material evidence by spectrofluorometry and fluorescent hemotest developed at Bureau for Forensic Medical Expert Evaluations of the Leningrad region and by traditional methods used in Russia and abroad is compared. The proposed methods are 1000 times more effective than the methods routinely used in Russia; moreover, they allow objective computer recording and processing. These methods are 50-70 times cheaper than the methods used in foreign countries and are virtually as sensitive. Fluorescent hemotest persuasively proves the presence of blood in stains on material evidence under laboratory conditions, at the site of accident, and even under field conditions (express analysis).     

大鼠死后心血中吗啡浓度变化的HPLC检测

本文采用高效液相色谱（HPLC）分析技术检测了治疗量及中毒量吗啡肌注大鼠死后心血中吗啡浓度变化．结果表明．以治疗量吗啡肌注大鼠死后96h内，心血中吗啡浓度随死后时间增加而显著升高（P<0.01），吗啡浓度水平与死后时间呈显著正相关．以中毒量吗啡肌注大鼠死后12h内，心血中吗啡浓度无明显变化．死后24h、48h及96h．随死后时间延长，心血中吗啡浓度逐渐升高（P＞0.01），其递增强度不如治疗量吗啡组大鼠的明显．本研究证实死后尸体心血吗啡浓度明显受生前剂量的影响，且死后96h内，随死后时间延长心血中吗啡浓度不断增高．本文初步探讨了死后心血吗啡浓度变化发生的可能机制．为海洛因或吗啡中毒死亡的血液检测结果评判及死因分析提供理论依据．     

Identification of menstrual blood by real time RT-PCR: technical improvements and the practical value of negative test results

The identification of body fluids can be of crucial importance in forensic casework. Recently we have shown that the detection of cell- and tissue-specific mRNAs is a suitable technique to identify menstrual blood and semen. In this paper we present an improved method using real time-PCR and the menstrual blood marker MMP-7 which increases sensitivity and specificity. We furthermore demonstrate case examples described showing that the correct identification of samples as non-menstrual is likely to be in the focus of criminal investigations. Although negative test results may have a number of other reasons, the new technique provides quantitative results thus allowing a cut-off value for the control marker GAPDH to be defined, below which negative results for the menstrual blood marker MMP-7 allow the conclusion that the sample was indeed not menstrual blood. Using these data, it is possible to apply the molecular identification of menstrual blood to forensic casework considering both the limitations and the capabilities of the method.     

Detection of diatoms in blood by a combination of membrane filtering and chemical digestion

An improved method for detecting diatom in blood is reported. Blood of cadavers was obtained by cardiac puncture at inquest or from the left atrium directly at autopsy. The blood was hemolyzed by sodium dodecyl sulfate (SDS) and filtered through membrane filter (47 mm in diameter and 5 microns in pore size). When the blood was putrefied, two or three pieces of membranes filters were needed because of choking membrane pore. The membrane filters were digested with fuming nitric acid and diluted with distilled water. The diluted solution was filtered through membrane filter (25 mm in diameter) again. After drying the membrane filter was immersed in oil. Diatoms on the membrane filter were clearly observed microscopically.     

固相萃取/气相色谱法检测全血中的扎来普隆

目的建立全血中扎来普隆的固相萃取/气相色谱检测方法。方法采用Oasis HLB固相萃取柱,5%甲醇水淋洗,二氯甲烷洗脱,GC/NPD定量检验,GC/MS定性检验,并对检测条件进行考察。结果扎来普隆与血中杂质分离良好,平均萃取回收率达89%以上,最低检出限20ng/mL,50～2000ng/mL内线性良好,相关系数R2=0.9945,日内RSD为3.1%～4.2%,日间RSD 5.8%～6.7%。结论该方法操作简便快速,提取回收率高,重现性好,提取物杂质少,可用于实际案件的检验。