大鼠死后心血吗啡浓度变化的HPLC检测

采用高效液相邑谱分析技术（HPLC）检测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，以治疗量吗啡肌往大鼠，在死后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里显著正相关；以中毒量吗啡肌注大鼠，在死后12h内，心血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，且在死后96h内，随死后时间增加．心血中吗啡浓度少数不断增高。     

The extent of postmortem drug redistribution in a rat model.

The aim of this study was to investigate the postmortem redistribution of several drugs in a rat model and to examine if any of the pharmacological properties was related to the extent of this phenomenon. One of the following drugs: phenobarbital (phenobarbitone), acetaminophen (paracetamol), carbamazepine, codeine, verapamil, amphetamine, mianserin, trimeprazine (alimemazine) or chloroquine was administered together with nortriptyline orally to rats 90 min prior to sacrifice. Heart blood was sampled immediately before sacrifice and after 2 h postmortem, as it has previously been shown that this is sufficient time for postmortem concentration changes to occur in heart blood. Blood was also sampled from the clamped abdominal inferior vena cava (representing peripheral blood) and tissue samples were taken from lungs, myocardium, liver, kidney, thigh muscle, forebrain, and vitreous humor together with a specimen from the minced carcass. Drugs were analyzed by high performance liquid or gas chromatography. For phenobarbital, acetaminophen and carbamazepine the postmortem to antemortem blood drug concentration ratios were close to 1.0 and tissue concentrations were low. The postmortem to antemortem heart blood drug concentration ratio for chloroquine (6.9 +/- 1.5) was higher than for nortriptyline (3.5 +/- 0.3), and the remaining drugs (codeine, verapamil, amphetamine, mianserin, and trimeprazine) showed ratios of the same magnitude as nortriptyline. The postmortem to antemortem blood drug concentration ratios for both heart blood and blood from the vena cava and also the lung to antemortem blood drug concentration ratio were closely related to the apparent volume of distribution for the drugs studied (p < 0.001). Accordingly, an apparent volume of distribution of more than 3-4 L/kg is a good predictor that a drug is liable to undergo postmortem redistribution with significant increments in blood levels. The postmortem drug concentration in blood from vena cava was closely related to the antemortem blood level, confirming that among the postmortem samples, the peripheral blood sample was the most representative for the antemortem blood concentration.     

Postmortem redistribution of digoxin in rats

Adult male Wistar rats were treated with either 0.1 or 3 mg/kg body weight X day of digoxin for five days, then killed and stored at 4 degrees C for 12 h in an attempt to mimic the normal preautopsy procedures in our hospital. In rats treated with 0.1 mg/kg body weight X day, the antemortem serum digoxin concentrations (SDC) were 1.1 +/- 0.4 ng/mL while the 12-h postmortem concentration was markedly increased (16.3 +/- 5.9 ng/mL) (P less than 0.01). In rats treated with 3 mg/kg body weight X day, SDC was not changed significantly (11.2 +/- 4.8 ng/mL antemortem and 13.3 +/- 6 ng/mL postmortem). Postmortem redistribution of digoxin was assessed by injection of 125I-labelled digoxin with or without pretreatment with the unlabelled drug. The results indicate that after death passive redistribution of digoxin may take place. When the SDC are within the therapeutic or low toxic range, digoxin may reenter the blood. High antemortem serum concentrations of digoxin may prevent such passive redistribution. Therefore, antemortem digoxin intoxication cannot be reliably inferred on the basis of high postmortem levels of the drug. Digoxin intoxication can be ruled out when postmortem SDC remain within the therapeutic range. The above changes cast doubt on some of the forensic and cardiologic literature, which has in the past been based on incorrect assumptions concerning postmortem behavior of digoxin.     

Postmortem metabolic indices of antemortem adrenergic stimulation

Tissue lactate concentration has been reported to be a useful postmortem indicator of antemortem awareness of mortal danger. The purpose of this study was to determine further whether selected tissue metabolites could be used as postmortem markers of antemortem adrenergic stress. Sprague-Dawley albino rats were anesthetized with pentobarbital and then injected with 2.0 mg kg-1 i.p. epinephrine hydrochloride to induce experimentally a severe sympathetic response that may be associated with the awareness of mortal danger; 20 min after the injection of epinephrine, when the metabolic response was at its peak, the animals were killed by exsanguination. Samples of the following tissues were removed immediately prior to death (0 h) and 48 h postmortem: soleus, plantaris, kidney medulla, kidney cortex, liver, and heart. These samples were analyzed for glycogen, lactate, ATP, creatine phosphate, pH, and total protein concentration. Significant differences in lactate concentration were observed in all tissues except soleus at 0 h in the epinephrine-injected animals. Specific tissues also had significant reductions in glycogen, ATP, and creatine phosphate concentrations at 0 h. At 48 h postmortem, however, only the liver and soleus lactate concentrations were significantly different from the 48-h control samples. It is unlikely that these small differences found in some tissues at 48 h postmortem would be detected in an uncontrolled accident situation. We concluded from these findings that these selected tissue metabolites are not useful as long-term postmortem indicators of antemortem adrenergically induced hypermetabolism.     

Timely antemortem and postmortem concentrations in a fatal carbamazepine overdose.

There are no published reports that include both timely antemortem and postmortem carbamazepine concentrations after massive overdose. We report a fatal overdose of carbamazepine with both timely antemortem and postmortem carbamazepine concentrations. Carbamazepine concentrations were 47.7 mcg/mL 2 h antemortem and 53 mcg/mL at 9 h postmortem. The slight rise in drug concentration may reflect continued absorption of the drug in the last 2 h before death. Postmortem carbamazepine concentrations drawn from a peripheral vessel in this patient appeared to reflect drug concentrations at the time of death.     

Investigation of cardiac troponins in postmortem subjects: comparing antemortem and postmortem levels

The limitations of autopsy in the diagnosis of death due to ischemic heart disease are well known. In the living, a simple reliable biochemical assay for cardiac troponins is used in the diagnosis of acute myocardial ischemia. Several studies have investigated the use of biochemical assays for cardiac troponins in postmortem subjects as a means to distinguish between a cardiac and anoncardiac cause of death. All of these studies, however, rely upon assigning subjects to "cardiac" or "noncardiac" death on the basis of a postmortem examination. As postmortem examination does not always accurately distinguish between these two groups, this approach is intrinsically flawed.Our study compares antemortem and postmortem cardiac troponin levels in five subjects. The antemortem samples were retrieved from the hospital biochemistry laboratory after each subject's death. The postmortem samples for each subject were taken from different sites and at different times during the early postmortem period.Erratic results bearing little or no relation to the antemortem cardiac troponin level were obtained for all subjects. Four of the five subjects had raised antemortem troponin levels, although only one had a cardiac cause of death.From this, we conclude that postmortem blood is not a suitable substrate for standard biochemical assays of cardiac troponins, which are designed for use on serum taken from living patients. In addition, the results of our study support the view that elevated cardiac troponins are a marker of serious morbidity and are not specific for cardiac injury as the primary cause of morbidity or mortality.     

Linearization of the relationship between postmortem plasma chloride concentration and postmortem interval in rats

A potentiometric titration procedure was employed for measurement of plasma chloride concentrations during the 0-96 h postmortem period in rats. The data revealed antemortem absolute values and postmortem rate of decrease in plasma chloride concentration (PCl) which were almost identical to those in dogs and in man. Expression of the data in the form of a double logarithmic plot of PCl versus postmortem interval (PMI) yielded a linear relationship of high correlation (r = -0.97; P less than 0.001). Attention is drawn to the possibility of utilizing postmortem rate of change in plasma chloride concentration as a means of estimating time elapsed since death.     

氯胺酮在急性中毒大鼠体内的死后再分布研究

目的探索氯胺酮在大鼠体内的死后再分布变化规律及温度对再分布的影响。方法48只雄性SD大鼠随机分为2个实验组（室温组24只、冷藏组18只）和1个对照组（6只）,实验组大鼠以氯胺酮290mg／kg灌胃,45min后缺氧处死,分别置于室温（24℃）和冷藏（4℃）条件下,于死后不同时间（0、12、24、48h）取心血、外周血、肝、肺、肾、心肌、大脑,检测其中氯胺酮含量;对照组大鼠以生理盐水灌胃,各对应组织器官样品为空白对照。血和组织样品中加入内标物SKF。。后碱化,乙酸乙酯萃取,GC／MS全扫描定性,内标法、工作曲线法气相色谱定量分析。结果室温条件下,大鼠死后48h内随着死亡时间延长,心血、肺、肝中氯胺酮的浓度呈升高趋势（P〈0．05）,肾脏中氯胺酮的浓度先升高后下降（P〈0．05）,外周血、心肌和脑中氯胺酮的浓度无显著性变化（P〉0．05）。冷藏条件下,血液及组织中氯胺酮浓度变化无显著性差异（P〉0．05）,除心肌外,各样本浓度均低于相应时段室温条件保存的样本。结论氯胺酮在大鼠体内存在死后再分布现象。温度对大鼠死后血液及组织中氯胺酮浓度变化有较明显的影响。     

References for determining the time of death by potassium in vitreous humor

The different statements concerning the slope and intercept of the regression line and the 95% limits of confidence are the reason that potassium in vitreous humor is not used (at least in Germany) as an aid in estimating the time of death. The relationship between the concentration of potassium and the time of death is mainly influenced by antemortem electrolyte imbalances caused by disease and/or duration of terminal episode. The influence of terminal episode is best identified by its duration (Adelson et al., J. Forensic Sci., 8 (1963) 503-514). In order to have a method suitable for every case and to be as precise as possible we looked therefore for parameters in vitreous humor which were stable postmortem and indicating antemortem electrolyte imbalance. Urea is such a parameter, being stable postmortem (Coe, Am. J. Clin. Pathol, 51 (1969) 741-750) and useful as a marker of antemortem electrolyte imbalance. Our investigations on potassium in vitreous humor, including sudden and hospital deaths after chronic lingering disease, revealed 95% limits of confidence of +/- 34 h up to 120 h postmortem. Reviewing only cases with urea less than 100 mg/dl the 95% limits of confidence could be reduced to +/- 22 h. Considering the duration of terminal episode (less than 6 h) the precision was +/- 20 h. In this way our modified procedure is suitable for every case with the resulting precision of estimation being determined only by the duration of the terminal episode and urea concentration.     

电击死兔骨骼肌及心肌HSP70 mRNA和c-fos mRNA表达

目的 研究电击死兔骨骼肌与心肌HSP70 mRNA和c-fos mRNA-表达变化。探究生前电击与死后电击的鉴别方法。方法 15只新西兰兔,随机分电击死组、死后电击组和对照组,每组5只,用荧光RT-PCR技术检测骨骼肌与心肌热休克蛋白70（HSP70） mRNA与c-fos mRNA表达水平,对所得结果进行统计学分析。结果 生前电击兔骨骼肌及心肌HSP70 mRNA与c-fos mRNA表达高于死后即刻电击者（P〈0．05）。结论 检测骨骼肌及心肌HSP70 mRNA与c-fos mRNA表达变化有助于于生前电击与死后电击的鉴别。     

缢死大鼠肝肾细胞内HIF-1α免疫组化染色观察

目的探讨缺氧诱导因子1α(HIF-1α)在窒息死亡鉴定中的意义。方法制作大鼠缢死后0、2、6、24h的窒息死模型,以相应时间段断颈处死大鼠为对照,用免疫组织化学ABC法结合图像分析观测肝和肾组织中的HIF-1α表达情况,并对其结果进行统计分析。结果除0h段外,HIF-1α免疫组化阳性染色可见于窒息组和对照组的各时段大鼠,主要位于肝细胞、肾近曲小管和远曲小管上皮细胞。死后6h内的肝组织HIF-1α免疫组化染色显示窒息组与对照组差别明显(P<0.05),24h后则无明显差别(P>0.05)。肾脏窒息组与对照组差别明显(P<0.05)。结论观测HIF-1α在死后6h内肝脏或24h内肾脏中的表达状态,对机械性窒息的鉴定有一定的法医学意义。     

扫描电子显微镜下生前和死后枪弹创的对比观察

本文用扫描电镜,对11例生前枪弹创和12例死后不同时间形成的枪弹创作对比观察。结果发现,生前枪弹创有大量红细胞和血小板凝块,纤维蛋白网形成,死后10min内的枪弹创亦有少许散在红细胞和纤维蛋白形成,很少见纤维蛋白网。死后25min的标本仍可见稀疏的红细胞,但无纤维蛋白形成。作者认为在扫描电镜下诊断生前枪弹创,需全面观察,综合分析。     

Temperature-correction of abdominal impedance: improved relationship between impedance and postmortem interval

The relationship between extracellular abdominal impedance and postmortem interval (PMI) reflects the combined effects, on impedance, of postmortem cooling of the tissues and of autolysis per se. This study was performed in order to eliminate temperature change as a major factor contributing to the time course of postmortem change in abdominal impedance. Dissociation of thermal and autolytic influences was achieved by recording deep abdominal temperature at the time of impedance measurement, followed by correction of all measured impedances to their theoretically predicted values at an arbitrarily chosen temperature of 40 degrees C. Uncorrected abdominal impedance increased from 82+/-12 Ohmz, 1 h after death, to 108+/-21 Ohmz after 12 h. Impedance then decreased to 96+/-23, 89+/-22, 75+/-19, 66+/-21 and 59+/-19 Ohmz at postmortem intervals of 24, 36, 48, 60 and 72 h, respectively. In contrast, corrected abdominal impedance decreased progressively from 63+/-7 Ohmz, 1 h after death, to 61+/-9, 56+/-11, 51+/-10, 46+/-10, 39+/-11 and 35+/-10 Ohmz at postmortem intervals of 12, 24, 36, 48, 60 and 72 h, respectively. The improved relationship between (corrected) abdominal impedance and PMI is of potential value in estimating time since death.     

Preliminary study of the effect of acute antemortem haemorrhage on postmortem abdominal impedance in rats

Studies over many years have revealed a consistent, inverse relationship between extracellular impedance of the rat abdomen and postmortem interval (PMI). Recent studies have shown that this relationship can be improved by correcting measured impedances to their theoretically predicted values at an arbitrarily chosen temperature of 40 degrees C, thus rendering them independent of the tissues' temperature at the time of impedance measurement. The present study, undertaken as a part of an ongoing effort to identify variables which might influence postmortem rate of change in abdominal impedance, was aimed at studying the possible effect of acute antemortem haemorrhage on abdominal impedance. Sudden loss of approximately 30% of the total blood volume, immediately prior to the death was without statistically significant effect on the pattern of postmortem change in abdominal impedance. Thus, in the control and experimental rats, respectively, impedance decreased progressively from 58.7 +/- 1.8 and 68.0 +/- 11.3 omega, 1 h postmortem, to 31.5 +/- 2.2 and 33.7 +/- 4.7 omega at a postmortem interval of 120 h (n = 6 in each group). In both groups, the relationship between impedance and postmortem interval was either linear or curvilinear. These findings are believed to mitigate in favour of continued effort to examine the potential usefulness of abdominal impedance, as an index of postmortem interval, under conditions encountered in routine forensic investigation.     

Cardiac blood pH as a possible indicator of postmortem interval

Postmortem changes in the pH of blood and selected tissues in rats were evaluated at intervals ranging from 2 min to 96 h. Cardiac blood pH was significantly and reproducibly decreased in all groups at all postmortem intervals, independent of the method of sacrifice used. A preliminary study using cardiac blood obtained at autopsy from a limited number (n = 11) of human subjects demonstrated a significant negative correlation (r = -0.908, P less than 0.01) between postmortem interval (range 2 to 20 h) and cardiac blood pH.     

大鼠百草枯中毒后体内的分布研究

目的建立生物检材中百草枯的超高效液相色谱-质谱联用检测方法,研究百草枯灌胃染毒致死的大鼠动物模型。方法大鼠以1/2 LD_(50)剂量灌胃染毒,分别于染毒后0.5h、2h、4h、8h、12h、24h、48h、72h处死解剖,采集心、肝、脾、肺、肾、脑、肌肉、膀胱和胃组织,UPLC-MS/MS法定量检测各组织中百草枯。结果试验中大鼠灌胃后,4h以内胃是主要分布器官,胃中含量最多,其他器官中含量相对较低。4h内除胃以外的脏器含量变化不大,4h后胃内百草枯含量有所下降,除胃以外的脏器含量均升高。各组织与脑组织比较具有显著性差异(P0.05)。结论百草枯在大鼠体内死后分布不均匀并且各组织含量随着时间变化有所改变。百草枯UPLC-MS/MS方法、口服染毒致死的动物模型、各组织分布规律可为甲百草枯中毒死亡案件提供检测依据。     

急性吗啡中毒大鼠主要脏器内吗啡分布变化的研究

研究急性吗啡中毒大鼠随给药和死后时间延长 ,主要脏器内吗啡的分布变化规律 ,为吗啡类毒品中毒死亡者尸检取材提供依据。采用免疫组织化学SP技术 ,观察 44只尾静脉注射吗啡的大鼠。从给药后 15min到 5h ,从死后即刻至 48h ,脑、肾、心、肝等脏器内吗啡分布的变化规律。结果表明 ,注射吗啡后短时间内各脏器均有吗啡存在 ,主要分布在某些实质细胞的胞浆内 ,且随时间延长吗啡含量上升 ,达高峰 ( 1h)后逐渐减少或消失。不同组织器官的吗啡含量及其变化速率差异巨大。脑内吗啡出现早 ( 15min) ,消失晚 ( 5h) ,峰值高 ,死后衰减慢 ( 4 8h仍呈强阳性 )。肾脏次之 ,但明显优于心、肝。免疫组化SP技术可作为一种鉴定吗啡类毒品中毒的特异性方法 ,脑、肾是其较理想的检材     

家兔死后体表锐器伤出血现象的研究

目的对家兔死后体表锐器损伤出血现象进行研究,以期能够贴近实际办案需要,找到一个更为实用的鉴别生前锐器伤和死后锐器伤的方法。方法家兔脱毛,制作锐器损伤模型,采用大体观察结合HE染色镜下组织病理学观察。结果死后锐器损伤出血量均较少,随着时间延长出血量减少,出血速度慢。死后30min的锐器损伤在形成过程与生前损伤有所区别,但在死后12h肉眼观察结果与生前损伤难以区别。死后1h以上的锐器损伤与生前损伤不同之处在于创缘不会被血染。结论位于尸体低下位置的死后30min内的锐器创与生前锐器创的区别是出血量相对较少。死后60min-90min的锐器伤出血量少,创缘皮肤不被血染,肌肉的出血较局限,与生前损伤相鉴别较容易。     

不同死后时间组织中吗啡免疫组化检测的初步研究

目的检测不同死后时间组织中的吗啡。方法应用抗吗啡兔血和SABC法对不同死后时间大鼠肝肾组织中吗啡进行了检测。结果初步发现死后27h仍可在肝肾组织普通石蜡切片上检出阳性染色。结论免疫组化技术可以用以检测死后组织中的毒物。     

Efficacy of cerebro-spinal fluid biochemistry in the diagnosis of brain insult.

Postmortem biochemical indices may provide a useful adjunct to morphological studies in the identification of antemortem brain insult. We studied 34 routine medico-legal cases categorising them into one of four diagnostic groups. There were 11 cases of head trauma, 7 of 'hypoxia' (3 hangings and 4 carbon monoxide or drug poisonings), 7 sudden cardiac deaths and 9 miscellaneous cases. Survival time and postmortem interval was known for each case. The degree of cranio-cerebral trauma was graded. Cerebro-spinal fluid (CSF) and vitreous humour were analysed for calcium, glucose, total proteins, aldolase, aspartate transaminase (AST), alanine transaminase (ALT), gamma glutamyltransferase (GGT), lactate dehydrogenase (LDH), creatine kinase (CK) and creatine kinase BB isoenzyme (CK-BB). CK-BB was also measured in superior vena cava serum. In CSF there was a significant correlation between the severity of cranio-cerebral trauma and levels of aldolase, CK-BB, AST, ALT and total proteins. CSF CK-BB, median units/l (range), for the groupings of head trauma, hypoxia, sudden cardiac death and miscellaneous were respectively 823 (2-3431); 96 (2-187); 4 (2-25); 5 (1-69). Corresponding serum CK-BB levels were 240 (28-322); 390 (26-411); 180 (20-482); 79 (18-530).