甲胺磷及其代谢产物检验的研究

本文利用ＧＣ／ＭＳ和ＧＣ／ＦＰＤ法测定了生物体内的甲胺磷（ＭＡＰ）及其代谢物Ｏ、Ｓ－二甲基磷酸酯（Ｏ、Ｓ－ＤＭＰ）．血中ＷＡＰ以乙、乙酸乙酯混合溶剂提取、以磷酸三丁酯为内标测定；尿液中Ｏ、Ｓ－二甲基磷酸酯（Ｏ、Ｓ－ＤＭＰ）以乙酸提取，以ＤＭＴＰ氘氢化合物为内标测定。ＭＡＰ最小检出量为１０ｎｇ／ｍｌ血，回收率为７５．１％，Ｏ、Ｓ－ＤＭＰ的最小检出量为２０ｎｇ／ｍｌ尿．     

GC／MSD分析生物样品中的巴比妥类药物

本文建立了ＧＣ／ＭＳＤ分析生物样品中五种巴比妥类药物的测定方法。线性范围是２００～５０００ｎｇ／ｍｌ，方法的回收率：巴比妥８１．４１±４．５０；异戊巴比妥８３．５８±６．６５；戊巴比妥８２．７８±７．１７；司可巴比妥７９．０４±７．２０；苯巴比妥８２．１３±６．８１（ｎ＝６）。     

尿样中海洛因代谢物的测定及海洛因滥用的确认

用ＳＰＥ－ＧＣ－ＮＰＤ法建立了尿样中吗啡、６－单乙酰吗啡及可待因的定性分析方法,适用于海洛因滥用者的尿样分析。尿样中吗啡及可待因的最小检测限均为５０ｎｇ／ｍｌ。方法的相对标准偏差分别为：吗啡１１．３％（ｎ＝５）,可待因１４．２％（ｎ＝５）。方法简便、灵敏、快速,１５ｍｉｎ可完成一例尿样的分析。研究了服用含可待因成分的复方甘草合剂后,尿样中的吗啡及可待因的峰面积比为０．４５７±０．１９７（Ｐ＝９９％）;统计了４０例明确滥用海洛因尿液的分析结果,吗啡与可待因的峰面积比为３．４６±０．８９４,Ｐ＝９９％。可作为判断海洛因滥用的依据。同时与免疫板法比较,附５５例免疫板法阳性尿样的分析结果     

大麻的体内代谢物THC—COOH的分析

本文介绍了定性定量分析大麻的主要代谢物ＴＨＣ－ＣＯＯＨ的两种方法。所有试管硅烷化，尿样碱性水解。ＴＬＣ法快速、简便，适用于多份检材的同时定性分析，最低检测限为５０ｎｇ．采用衍生化、ＳＩＭ的ＧＣ／ＭＳ方法灵敏，选择性、专一性强．五氟丙酸酐衍生化方法线性范围为２０－２５０ｎｇ／ｍｌ，最低检测限１０ｎｇ／ｍｌ，变异系数为８．４％，提取回收率为５８．９％；甲基化方法线性范围１０～１００ｎｇ／ｍｌ，最低检测限５ｎｇ／ｍｌ，变异系数为５．６％，提取回收率为５３．２％．     

SPME-GC\NPD法快速分析尿液中苯丙胺类化合物

建立了ＳＰＭＥ－ＧＣ／ＮＰＤ法同时分析尿液中九种苯丙胺类化合物的方法。取１ｍｌ尿样以４－苯基丁胺为内标,在碱性条件下,将１００μｍＰＤＭＳ纤维头浸入尿液中２０ｍｉｎ,气相色谱进样后热解吸３ｍｉｎ,用ＧＣ／ＮＰＤ进行测定。九种苯丙胺类化合物及内标完全分离,五种苯丙胺类化合物分别在０．０５～１５、０．１～１５、０．２～１５μｇ／ｍｌ浓度范围内线性良好,ｒ为０．９９２８～０．９９９５,变异系数均小于１０％。方法简便、快速、准确,可用于尿液中苯丙胺类违禁药物的检测     

酒精对乌头碱毒物动力学的影响

用反相高效液相色谱法测定生物样品中的乌头碱．给新西兰兔灌服１．５ｇ／ｋｇ短柄乌头后，乌头碱在免休内的毒物动力学过程为二室开放模型，其Ｋａ、Ａ、Ｂ值分别为１．１６２９±０．４０５３，０．６０４６±０．２５７４，１．１６０７±０．３７８１ｕｇ／ｍｌ．研究了灌服５．７５ｇ酒精对该动力学过程的影响，表明酒精不改变其模型类型，但可明显加速乌头碱的吸收和分布（Ｐ＜０．０１），对消除过程无明显影响，其Ｋａ、Ａ、Ｂ值分别为２．４０２６±０．５３７６，１．２０５１±０．５３２８，１．２０３７±０．４０９５μｇ／ｍｌ。说明酒精可增加短柄乌头的毒性。     

车祸引起失血性休克致右眼前段缺血性视神经病

王某,男,２１岁。于１９９８年２月２０日在成都郊区公路上骑摩托车时,与一面包车相撞,王伤后立即送至成都某医院就诊。查体：神志不清,ＢＰ５～３ｋＰａ,脉搏１３０次／分,微弱,面色苍白,四肢湿冷,双瞳孔直径０．５ｃｍ,对光反射迟纯,头颅未见异常,颈软,右胸前青紫６ｃｍ×８ｃｍ,腹稍隆,右上腹腹肌紧张,右下肢青紫肿胀４ｃｍ×５ｃｍ,头部ＣＴ示颅内未见异常,胸部Ｘ光摄片示右胸腔积血,左肋骨５～９肋骨折,右下肢Ｘ光摄片示右胫、腓骨中段粉碎性骨折。立即给予输血、输液,行剖腹探查见腹腔积血１０００ｍｌ,肝破裂…     

星状神经节麻醉诱发急性心衰猝死1例

案例　某男,５７岁。主因颈部及左肩部疼痛到某门诊部就治。查体：Ｐ８０次／分,ＢＰ１２０／８０ｍｍＨｇ,颈部活动受限,第６、７颈椎有压痛;左肩关节压痛明显,活动受限。问无心脏病史。诊断为颈椎病,左肩周炎。采用局部封闭治疗,用药为利多卡因０．１ｇ及骨宁５ｍｌ,强的松龙２．５ｍｌ。用９号注射针头先在左肩关节两处压痛点各半量注射,观察１５分钟无异常。随即用上述药物在第７颈椎旁各半量注射,约１０分钟后,患者出现胸闷、心慌、头晕、血压急剧下降,经急救２小时无效死亡。尸检主要所见：成年男尸,发育正常。角膜轻度…     

胸部刺创致胸腹多脏器损伤1例

胸部刺创致胸腹多脏器损伤１例苏凤生张某，男，１５岁，学生。伤者在溜冰时与他人发生口角，后被用刀捅伤。急送医院抢救。入院查体：Ｔ３６Ｃ，Ｐ８０次／分，Ｒ２０次／分，ＢＰ９／５Ｋｐａ。口唇苍白，四肢偏冷，神志模糊，左胸部第八肋间有一５ｃｍ创口，深达胸腔，...     

大鼠脑损伤后GDNF表达变化的时间规律性研究

目的探讨大鼠脑损伤后胶质源性神经营养因子(GDNF)在神经元、神经胶质细胞中表达变化的时间规律性。方法以冲击应力σd为355.09kPa致大鼠脑损伤后,于不同时问段分别应用原位杂交、免疫组化、免疫组化双染色技术检测GDNF mRNA和蛋白的表达强度,用图像分析系统测定阳性反应物平均灰度,进行统计学分析。结果正常对照组GDNF mRNA和蛋白弱表达,伤后15min开始表达呈增强趋势,分别于伤后1d、2d,表达至峰值,平均灰度值分别为133.08±4.53、125.05±2.00,与对照组、邻近上组比较均具有显著性差异(P<0.05),并维持高水平表达至伤后7d,平均灰度值分别为116.65±1.31、114.20±1.68,与对照组比较具有显著性差异(P<0.05)。表达至峰值前,阳性细胞多为神经元,之后以神经胶质细胞为主。结论脑损伤后GDNF mRNA及蛋白的表达具有时间规律性,GDNF阳性细胞种类存在变化,可作为法医学推断脑损伤形成时间的指标。     

The influence of sex hormones on the elimination kinetics of ethanol

The goal of the investigation was to research the influence of sex hormones on the elimination kinetics of ethanol. Forty-seven healthy men (average age 25+/-6.1 years) and 61 healthy women (average age 24+/-2.4 years) received 0.79-0.95g of ethanol/kg body weight in the form of an alcohol beverage of their choice. The target concentration for both sexes was a blood alcohol concentration (BAC) of 1.10g/kg. Blood samples for the determination of the ethanol concentration followed in the elimination phase in 10-20min intervals. The sex hormone levels (estradiol, progesterone and testosterone) were determined concomitantly from the serum. In men, the mean testosterone concentration was 5.3+/-1.6ng/ml, the mean estradiol concentration was 34.6+/-13.6pg/ml and the mean progesterone concentration was 0.9+/-0.3ng/ml. In women, the mean estradiol concentration was 47.6+/-52.6pg/ml and the mean testosterone concentration was 0.8+/-0.4ng/ml. Progesterone displayed a so-called dummy effect in women. In the high progesterone group (n=11), the mean concentration was 11.1+/-3.5ng/ml and in the low progesterone group (n=50) the mean was 0.6+/-0.3ng/ml. The mean hourly elimination rate (beta60) was 0.1677+/-0.0311g/kg/h in men. In women, the mean hourly elimination rate was 0.2044+/-0.0414g/kg/h in the high progesterone group and 0.1850+/-0.0276g/kg/h in the low progesterone group (p<0.05). The beta60 for women in the low progesterone group was significantly higher than that of the men, whose progesterone levels fell within a similar range (p>0.01). These results allow one to conclude that the gender differences in the pharmacokinetics of ethanol can partly, but not completely, be explained by progesterone levels.     

A sandwich enzyme immunoassay for pulmonary surfactant protein D and measurement of its blood levels in drowning victims

A sensitive sandwich enzyme immunoassay for human pulmonary surfactant protein D (SP-D) was developed and used to examine the blood SP-D levels of drowning victims. Human SP-D was purified from amniotic fluid by chromatographic methods, and an antibody against human SP-D was prepared. A polystyrene ball coated with anti-SP-D IgG was incubated with purified human SP-D, and then with anti-SP-D Fab'-peroxidase conjugate. Peroxidase activity bound to the polystyrene ball was assayed by fluorometry using 3-(4-hydroxyphenyl)propionic acid as the hydrogen donor. The detection limit of human SP-D was 5.2 pg per assay tube. Examination of cross-reactions of this sandwich enzyme immunoassay with proteins from other human organs showed it to be highly specific for lung, and Northern blot analysis detected specific SP-D mRNA expression only in lung. The SP-D concentration of normal human serum was 6.4+/-2.7 (mean+/-S.D.) ng ml(-1) (n=20). The recovery rates of 0.52 ng and 5.2 ng SP-D added to 5 microl normal human serum were 93.6+/-2.7% and 93.6+/-6.1%, respectively. Blood SP-D levels of victims from the saltwater drowning group (n=14) revealed higher concentrations (105.8+/-53.7 ng ml(-1)), while freshwater drowning victims (n=12) were estimated to be 74.1+/-43.9 ng ml(-1). The SP-D levels of 15 subjects who died of hemorrhage (n=5), heart failure (n=8), traumatic shock (n=1), and electrocution (n=1) were lower (22.0+/-8.5 ng ml(-1)), and those of asphyxia victims (n=10) were slightly higher (36.2+/-17.1 ng ml(-1)) than those of other causes of death, except for drowning. These results suggest that in drowning victims, SP-D flowed into the systemic circulation by physiological and physical mechanisms, and the differences of blood SP-D levels between saltwater drowning and freshwater drowning victims are presumed to be influenced by the type of agony and/or the length of survival time in water.     

Nicotine and cotinine levels in body fluids of smokers who committed suicide

Cigarette smoking is associated with a higher risk for suicide. The present study was conducted on the hypothesis that suicide smokers show higher nicotine and cotinine levels in blood and urine than non-suicide smokers. We determined nicotine and cotinine levels in blood and urine of 87 deceased individuals (18 suicides and 69 non-suicides) by gas chromatography. The smoking rate was 77.8% for individuals who committed suicide and 42.0% for those who did not commit suicide. Average nicotine and cotinine levels in blood were significantly higher in the suicide smokers than in the non-suicide smokers (nicotine: 93.2+/-46.6 ng/ml versus 25.8+/-14.4 ng/ml, p<0.0001 and cotinine: 378+/-235 ng/ml versus 201+/-137 ng/ml, p<0.005). Average levels of urinary nicotine and cotinine were also significantly higher in the suicide smokers than in the non-suicide smokers (nicotine: 1980+/-2210 ng/ml versus 394+/-376 ng/ml, p<0.005 and cotinine: 1170+/-1330 ng/ml versus 414+/-290 ng/ml, p<0.05). Twenty-six decedents were intoxicated with alcohol, and they included 7 suicides (7 smokers) and 19 non-suicides (15 smokers). Our data suggest that cigarette smokers who commit suicide smoke more heavily than other cigarette smokers.     

Postmortem serotonin (5-HT) concentrations in the cerebrospinal fluid of medicolegal cases

In a medicolegal study the postmortem serotonin (5-HT) cerebrospinal fluid (CSF) concentrations were determined in routine autopsies using a high performance liquid chromatographic procedure with electrochemical detection. There was no correlation between 5-HT concentrations and age, sex or blood alcohol concentration using a postmortem delay < or = 3 days. In suicides the suboccipital CSF concentrations were significantly decreased compared to the levels measured in the control group (8.55+/-5.99 ng/ml versus 20.15+/-13.56 ng/ml). Additionally, a decrease of 5-HT was found in the suboccipital CSF of opiate fatalities (15.56+/-13.52 ng/ml). The results support the hypothesis that decreased 5-HT concentrations in the CSF are characteristic in suicides. However, due to a rather broad overlapping of values between suicides and controls the results failed to define a possible cut-off level in the 5-HT CSF concentration to distinguish between a suicidal and a non-suicidal incident.     

Enzyme-linked immunosorbent assay for determination of plasma thyroglobulin and its application to post-mortem diagnosis of mechanical asphyxia

A high sensitive enzyme-linked immunosorbent assay (ELISA) for determination of plasma thyroglobulin was devised and applied to the postmortem diagnosis of mechanical asphyxia. The sensitivity of the present ELISA was 5 ng/ml. All of the 15 victims who died from causes other than mechanical asphyxia showed plasma thyroglobulin levels lower than 200 ng/ml (99.8 +/- 37.9 ng/ml), while 12 out of 14 victims of mechanical asphyxia showed the levels higher than 200 ng/ml (2100 +/- 3450 ng/ml in 14 victims). Thus, the present ELISA appears to be useful for postmortem diagnosis of mechanical asphyxia.     

Fast confirmation of 11-nor-9-carboxy-Delta(9)-tetrahydrocannabinol (THC-COOH) in urine by LC/MS/MS using negative atmospheric-pressure chemical ionisation (APCI)

A fast method using automated solid-phase extraction (SPE) and short-column liquid-chromatography coupled to tandem mass-spectrometry (LC/MS/MS) with negative atmospheric-pressure chemical ionisation (APCI) has been developed for the confirmation of 11-nor-9-carboxy-Delta(9)-tetrahydrocannabinol (THC-COOH) in urine samples. This highly specific method which combines chromatographic separation and MS/MS-analysis can be used for the confirmation of positive immunoassay results with a NIDA cut-off of 15ng/ml. The conjugates of THC-COOH were hydrolysed prior to SPE, and a standard SPE was performed using C18-SPE columns. No derivatisation of the extracts was needed as in GC/MS analysis, and the LC run-time was 6.5min by gradient elution with a retention time of 2.4min. Linearity of calibration was obtained in the range between 0 and 500ng/ml (correlation coefficient R(2)=0.998). Using linear regression (0-50ng/ml) the limit of detection (LOD) was 2.0ng/ml and the limit of quantitation (LOQ) was 5.1ng/ml; day-to-day reproducibility and precision were tested at 15 and 250ng/ml and were 13.4ng/ml+/-3.3% and 255.8ng/ml+/-4.5%, respectively.     

Urinary catecholamines in hyperthermia-related deaths

A group of five hyperthermia-related deaths is presented in which urinary noradrenaline (NA) concentrations were elevated (172.1 +/- 119.4 ng/ml) compared with a control group of rapid violent deaths (43.7 +/- 26.1 ng/ml). Urinary adrenaline (A) concentrations were not elevated in the hyperthermia cases, nor were there any significant differences in urinary dopamine (DA) concentrations between the two groups. All except one of the hyperthermia victims were under the influence of ethanol. It is suggested that a combination of heat stress and ethanol consumption was responsible for the elevated urinary NA in the hyperthermia cases, reflecting increased sympathetic nervous system activity. A combination of high urinary NA with low A seems to be characteristic of hyperthermia fatalities, in contrast to hypothermia deaths, where both NA and A are usually elevated.     

Concentration of drugs in blood of suspected impaired drivers

Analytical records concerning 440 living drivers suspected of driving under the influence of drug (DUID) were collected and examined during a 2 years period ranging from 2002 to 2003 in canton de Vaud, Valais, Jura and Fribourg (Switzerland). This study included 400 men (91%) and 40 women (9%). The average age of the drivers was 28+/-10 years (minimum 16 and maximum 81). One or more psychoactive drugs were found in 89% of blood samples. Half of cases (223 of 440, 50.7%) involved consumption of mixtures (from 2 to 6) of psychoactive drugs. The most commonly detected drugs in whole blood were cannabinoids (59%), ethanol (46%), benzodiazepines (13%), cocaine (13%), amphetamines (9%), opiates (9%) and methadone (7%). Among these 440 cases, 11-carboxy-THC (THCCOOH) was found in 59% (median 25 ng/ml (1-215 ng/ml)), Delta(9)-tetrahydrocannabinol (THC) in 53% (median 3 ng/ml (1-35 ng/ml)), ethanol in 46% (median 1.19 g/kg (0.14-2.95 g/kg)), benzoylecgonine in 13% (median 250 ng/ml (29-2430 ng/ml)), free morphine in 7% (median 10 ng/ml (1-111 ng/ml)), methadone in 7% (median 110 ng/ml (27-850 ng/ml)), 3,4-methylenedioxymethamphetamine (MDMA) in 6% (median 218 ng/ml (10-2480 ng/ml)), nordiazepam in 5% (median 305 ng/ml (30-1560 ng/ml)), free codeine in 5% (median 5 ng/ml (1-13 ng/ml)), midazolam in 5% (median 44 ng/ml (20-250 ng/ml)), cocaine in 5% (median 50 ng/ml (15-560 ng/ml)), amphetamine in 4% (median 54 ng/ml (10-183 ng/ml)), diazepam in 2% (median 200 ng/ml (80-630 ng/ml)) and oxazepam in 2% (median 230 ng/ml (165-3830 ng/ml)). Other drugs, such as lorazepam, zolpidem, mirtazapine, methaqualone, were found in less than 1% of the cases.     

Enzyme-linked immunoabsorbent assay for plasma thyroglobulin following compression of the neck

A highly sensitive enzyme-linked immunosorbent assay (ELISA) for determination of plasma thyroglobulin (Tg) which we devised was applied to the postmortem diagnosis of external compression of the neck. The coefficients of variation within and between assays were 2.4-6.6% and 6.8-12.0%, respectively. A significant correlation was observed between Tg levels measured by our ELISA and those measured by radioimmunoassay (r = 0.996, P less than 0.001). Plasma Tg levels in all 36 cadavers without external compression of the neck or neck injuries were lower than 200 ng/ml (73.6 +/- 51.9 ng/ml, M +/- S.D.) although these levels were a little higher than those in living bodies (16.7 +/- 11.8 ng/ml). On the other hand, plasma Tg levels in most of 42 victims of asphyxia due to external compression of the neck were higher than 200 ng/ml (2190 +/- 4300 ng/ml), and the highest one was 24,600 ng/ml. Plasma levels of the other thyroid hormones and thyroid-stimulating hormone were determined in some cases, and the cervical conditions in certain cases of strangulation were analyzed. Present results suggest that Tg was released by mechanical force on the thyroid gland added at the agonal stage. The determination of plasma Tg level in a cadaver using our ELISA would seem to be useful for the postmortem diagnosis of the presence of external compression of the neck. Although thyroid diseases causing elevated Tg levels are rare in forensic cases, they should be excluded by routine histology of the thyroid at autopsy, so that the finding of a high Tg level would thus carry more weight when given in evidence.     

Area-specific increased density of mu-opioid receptor immunoreactive neurons in the cerebral cortex of drug-related fatalities

In animal experiments and in cell culture, chronic morphine treatment has been followed by "up-regulation" as well as "down-regulation" of the mu-opioid receptor (OR) number. The present postmortem morphometric study of morphine-related fatalities of drug-addicts (n=13, 20-35 years old, with blood unconjugated morphine levels from 27.1 ng/ml to 458 ng/ml, m.v. 198.5 ng/ml) versus a non-addicted control group (n=13, 10-44 years old) was intended to examine, whether chronic opiate exposure affects the numerical density of mu-OR expressing neurons in the human neocortex (areas 11, 24 and 25 according to Brodmann). For the immunohistochemical procedure, vibratome sections (100 microm) were incubated with a monoclonal antibody against the mu-OR, diluted 1:100, and immunolabelled sites were visualized using an immunoperoxidase protocol. The numerical densities of OR immunoreactive neuronal profiles and Nissl-stained central profiles were assessed morphometrically (camera lucida-drawings). In both groups, the anti-mu-OR-immunoreactivity was mainly localized in pyramidal neurons of layers (L) II/III and V and in multiform neurons of L VI. In the areas 24 and 25, the density of the immunoreactive neuronal profiles did not display a significant difference between the two examined groups. In the area 11, however, the number of immunolabelled neuronal profiles amounted to 2777+/-206 mm(3) in the drug-related fatalities and to 2320+/-124 mm(3) in the control group and thus was significantly increased.