超生反应阶段骨骼肌机械性损伤的形态学研究

对20例1．25～8．30hpm具有超生反应能力的尸检骨骼肌和17例10～24hpm和48～60hpm不具超生反应能力的尸检骨骼肌进行机械性损伤刺激的形态学系列对比研究。结果表明，经机械打击刺激后，具超生兴奋性的肌肉，受打击部位的肌纤维出现类似典型生前伤征象的形态学改变，而不具超生兴奋性的肌纤维仅出现被动的肌纤维断裂等非活性肌纤维改变。通过对超生形态学反应的形成机理进行探讨，为进一步研究超生形态学的作用提示了前景。     

Electrical excitability of skeletal muscle postmortem in casework

The paper gives a review of the development of methods of postmortem electrical excitability of skeletal muscle for determining the time since death. For practical application in casework, the following method can be proposed: position of electrodes in the orbicularis oculi muscle, stimulation by rectangular impulses of 10 ms duration, 30 mA in a repetition rate of 50/s. The muscular reaction on excitation is graded in 6 degrees according to the spread of movement and the intensity of contraction. To each degree a time information is corresponding (95%-limits of confidence). These 95%-limits of confidence were proved to be valid for cases of sudden natural or traumatic deaths. Exclusions: deaths after chronic lingering disease (shorter times), fatal hypothermia, haematomas, emphysemas of the eyelid (longer times). The practical application in casework together with the temperature method (Henssge 1988, 1) is described. Applying both methods the death time estimation in the early postmortem interval may be much more precise and accurate than using one method alone.     

Enzymehistochemical and immunohistological changes of skeletal muscle motor end-plates and muscle fibers and their relation to the time of death

In order to investigate the relationship between the retrograde changes of the skeletal muscle and the time of death in various postmortem intervals (PMI), a systemic study of the enzymehistochemical activity of AChE, SDH, LDH, Ca(2+)-ATPase and the immunohistochemical reaction of SYN in motor end-plates and muscle fibers was conducted in rats under different temperatures and at various PMI. The results were analyzed and compared by an image processing system. It was found that these changes were related to the PMI, especially AChE changes. The AChE could be used as a sign-enzyme of skeletal muscle to date death.     

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.     

The Achilles tendon as a DNA source for STR typing of highly decayed corpses

Forensic criminal casework often involves DNA profiling of human postmortem tissues, whereas degradational processes can affect PCR-based Short Tandem Repeat (STR) analysis. Degradation of DNA is observed to vary among different tissues and with time. Therefore, the stability of DNA in Achilles tendon samples is compared to that in muscle and kidney specimens with a variety of postmortem histories. Tissue samples from 28 autopsy cases, including 15 decomposed corpses and a control group of 13 nondecayed corpses were analysed. DNA was isolated using the All-tissue DNA Kit (GEN-IAL, Troisdorf, Germany), quantified by spectrophotometric measurement, amplified by the multiplex PCR genRES MPX-2 (Serac, Bad Homburg, Germany), and analysed on the ABI PRISM 310 Genetic Analyzer (Applied Biosystems, Darmstadt, Germany). Quantitative analysis of nondecomposed tissues revealed that the recovery of DNA was highest in kidney followed by muscle, whereas Achilles tendon tissue was the poorest source of isolated DNA. Only small amounts of DNA were present in both kidney and muscle samples from decomposed corpses. However, from decayed Achilles tendon samples twice as much DNA as from nondecayed samples could be isolated on average. These results suggest DNA to be better protected in Achilles tendons. Moreover, postmortem changes in Achilles tendons may even improve DNA isolation.     

Spontaneous degeneration of the posticus muscle

The intrinsic muscles of the larynx were prepared in 265 forensic postmortems (death by strangulation: 73; other violent deaths: 93, natural death: 79, others: 20) for a histological examination. Muscle-fiber alterations were found in the (macroscopically normal) PCA muscle in most cases where the persons were of middle age or older. There were abnormalities such as rows or clumps of nuclei (also inside muscle fibers), the accumulation of lipofuscin pigment, focal or total necrosis of muscle fibers with or without predominant histiocytic infiltration, atrophic muscle cells, and fibrosis. The changes were mostly mild (normal muscle cells predominated) but clearly visible. If there had been acute damage to the larynx (for instance, death some time after attempted strangulation), these preexistent alterations must be taken into consideration when interpreting the microscopical findings.     

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。     

Study of elastic fiber changes of human skin in distinction between antemortem and postmortem wounds

Elastic fiber changes of volunteers' antemortem and postmortem skin wounds of various time were observed in order to find possible differences between them. These sections of the wounds were stained by Hart's modification of Weigert's elastic tissue stain.. However, this study showed that there were no differences in the nature and distribution of the elastic fibers in the dermis of volunteers' antemortem and postmortem human skin wounds. Therefore, this suggests that the appearances of the elastic fibers in the dermis should not be as a means of differentiating antemortem skin wounds from postmortem wounds in forensic identification.     

大鼠死后腓肠肌直接电兴奋性变化与早期死亡时间的关系

目的探讨大鼠死亡后不同时间腓肠肌电兴奋性的变化,为早期死亡时间的推断提供新的指标。方法建立大鼠机械性窒息模型,利用电生理检测仪检测死亡后不同时间大鼠腓肠肌对不同强度的电刺激的反应性,记录肌肉组织动作电位出现时间,统计学分析其差异并建立回归方程。结果大鼠机械性窒息死亡后一定时间内,其腓肠肌对电刺激的兴奋性与死亡时间呈明显线性关系。结论肌肉组织的电兴奋性可以作为早期死亡时间推断的有效指标。     

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.     

免疫印迹检测人胸大肌肌钙蛋白I推断较晚期死后经过时间

目的研究人死后骨骼肌肌钙蛋白I(Skeletal troponn in I,sTnI)的变化规律,探讨较晚期死后经过时间推断方法。方法以人胸大肌为研究对象,利用免疫印迹(W estern b lot)结合图象分析技术半定量检测不同离体时间内人胸大肌sTnI的含量,观察其与离体时间的关系。结果人胸大肌sTnI含量随离体时间延长而逐渐下降,与离体时间的对数值呈近似的线性关系:Y=11 972.5-4 761.9 lgX,相关系数r=0.989;离体5d的人胸大肌仍可检出sTnI。结论检验人死后骨骼肌(胸大肌)sTnI的含量有望成为用于推断人体较晚期死后经过时间的新技术。     

Supravital electrical excitability of muscles

A new method of postmortem electrical stimulation of muscles, measuring the force of the reaction against time, is presented. Preliminary investigations have yielded the following results: The maximum force of reaction in response to a definitive stimulation, measured by a sensitive force transducer, decreases in proportion to the post mortem interval. Using this method, the mechanical reactions of muscles by electrical stimulation can be demonstrated over a much longer postmortem period as these movements are visible. The duration of postmortem reaction of different muscles seems to be much more uniform than was previously assumed. Occasionally, reactions of the M. orbicularis oculi can be seen for the longest time. The basal muscular tonus changes relative to time. The time course of a single force reaction also changes relative to the postmortal interval: the reaction becomes slower. It has been theoretically established that measurement of influencing factors such as temperature and pH in the tested muscle will enable the time of death to be more precisely estimated.     

Acidosis, lactate, electrolytes, muscle enzymes, and other factors in the blood of Sus scrofa following repeated TASER exposures

Repeated exposure to electro-muscular incapacitating devices could result in repetitive, sustained muscle contraction, with little or no muscle recovery period. Therefore, rhabdomyolysis and other physiological responses, including acidosis, hyperkalaemia, and altered levels of muscle enzymes in the blood, would be likely to occur. Experiments were performed to investigate effects of repeated exposures of TASER International's Advanced TASER X26 on muscle contraction and resultant changes in blood factors in an anaesthetized swine model. A total of 10 animals were used. Six swine were exposed for 5 s, followed by a 5-s period of no exposure, repeatedly for 3 min. (In five of the animals, after a 1-h delay, a second 3-min exposure period was added.) The remaining four animals were used for an additional pilot study. All four limbs of each animal exhibited contraction even though the electrodes were positioned in areas at some distances from the limbs. The degree of muscle contraction generated during the second exposure period was significantly lower than that in the first exposure series. This finding was consistent with previous studies showing that prolonged activity in skeletal muscle will eventually result in a decline of force production. There were some similarities in blood sample changes in the current experiments with previous studies of muscular exercise. Thus problems concerning biological effects of repeated TASER exposures may be related, not directly to the "electric output" per se, but rather to the resulting contraction of muscles (and related interruption of respiration) and subsequent sequelae. Transient increases in hematocrit, potassium, and sodium were consistent with previous reports in the literature dealing with studies of muscle stimulation or exercise. It is doubtful that these short-term elevations would have any serious health consequences in a healthy individual. Blood pH was significantly decreased for 1h following exposure, but subsequently returned toward a normal level. Leg muscle contractions and decreases in respiration each appeared to contribute to the acidosis. Lactate was highly elevated, with a slow return (time course greater than 1 h) to baseline. Other investigators have reported profound metabolic acidosis during restraint-associated cardiac arrest. Since restraint often occurs immediately after TASER exposure, this issue should be considered in further development of deployment concepts. On the basis of the results of the current studies, the repeated use of electro-muscular incapacitating devices in a short period of time is, at least, feasible, with the caveat that some medical monitoring of subjects may be required (to observe factors such as lactate and acidosis).     

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。     

Practical experiences with the determination of cadaver age by evaluation of bacterial metabolic products

In an experiment a corpse had been kept at room temperature (16 degrees-23 degrees C) for 1163 h. At regular intervals brain samples were taken and the content of free amino acids and related compounds was determined by column chromatography. It could be demonstrated that in a period of 4-20 days postmortem the age of the corpse could be calculated from the concentrations of alpha-aminobutyrate (ABU), gamma-aminobutyrate (GABA), and glutamic acid (GLU) in brain employing the previously [5] presented formula: (formula; see text) T is the postmortem time lapse (days). ABU, GABA, and GLU are the concentrations (mumol/g) wet tissue of the corresponding amino acids. During this postmortem interval there is nearly a linear correlation; from this interval a correct assignment between concentrations and time cannot be given. The determination method is in the range of about 15 degrees-25 degrees C (guarantees bacterial growth and metabolism) independent of ambient temperature. The brain samples (cortex or parts of the putrified brain mush) can be taken without any special precaution during normal autopsy. The results of the experiment (in Fig. 1) were supported by some practical cases where the time lapse since death was well (in Fig. 1) or reasonably (in Fig. 1) known. These results suggest that this method allows in many cases the determination of the age of a corpse found in a warm environment (approximately 15 degrees-25 degrees C) approximately 4-20 days after death.     

不同环境条件下大鼠死后皮肤大体和组织学改变

目的研究观察大鼠在不同环境条件下死后不同时间其皮肤大体、组织学的改变,为推测较长的死后间隔时间建立实验动物依据。方法观察雌性SD大鼠在不同环境条件下的死后变化,并间隔一定死后时间取皮、染色和镜下观察。结果死后发生骨、软组织分离,A组约需35d,B组需13d。皮肤有形细胞成分和附属器完全分解破坏,A组约20d,B组7d。胶原纤维在A组死后40d（B组12d）仍有部分保留完整。结论环境温度差异是造成A、B两组发生死后改变时程不同的主要因素,利用皮肤大体、组织学改变可望较系统和准确的推测晚期死亡时间。     

Interleukin-6 and C-reactive protein serum levels in sepsis-related fatalities during the early postmortem period

Postmortem interleukin-6 (IL-6) and C-reactive protein (CRP) serum levels were investigated prospectively in sepsis-related fatalities and non-septic fatalities by using a linear regression model. At least three blood samples were collected between 0.3 and 139 h postmortem from sepsis-related fatalities (n=8) and non-septic fatalities (n=16). In addition, one antemortem blood sample was collected shortly before death from the septic patients. Antemortem and postmortem IL-6 and CRP levels were highly elevated in all individuals included in the sepsis group. An excessive postmortem increase of IL-6 serum levels associated with progressive time after death was observed in five out of the eight septic patients. Both, IL-6 and CRP serum concentrations seem to be suitable biochemical postmortem markers of sepsis. The determination of IL-6 serum levels above 1500 pg/ml in peripheral venous blood obtained in the early postmortem interval can be considered as a diagnostic hint towards an underlying septic condition. A more precise postmortem discrimination between sepsis and non-septic underlying causes of death is provided by the postmortem measurement of serum CRP in peripheral venous blood: on condition that at least two postmortem CRP values have been determined at different time points postmortem, the CRP level of a deceased at the time of death can be calculated by using linear regression analysis. When assessing postmortem IL-6 and CRP concentrations as biochemical postmortem markers of sepsis, various clinical conditions, such as a preceding trauma or burn injury going along with elevated IL-6 and/or CRP levels prior to death as a result of the systemic inflammatory response syndrome (SIRS) should be taken into consideration, thus adding relevant information for the practical interpretation of the results.     

Distribution of ethyl glucuronide in rib bone marrow, other tissues and body liquids as proof of alcohol consumption before death

Postmortem ethyl glucuronide (EtG) concentrations in rib bone marrow, liver, muscle, fat tissue, urine, blood and bile have been determined by LC-MS/MS. Samples have been taken from twelve corpses during autopsies. In nine corpses EtG could be detected, corresponding blood ethanol concentrations (BAC) were 0.04-0.37 g%. In three cases, no EtG was found; two of these cases showed postmortem BACs - possibly due to putrefaction - of 0.01 and 0.1g%. In rib bone marrow, which is easily accessible during autopsy, EtG concentrations (0.77-9.36 microg/g) have been lower than in blood (2.24-20.46 microg/mL) in eight of nine cases and comparable or higher than in muscle tissue. Therefore, rib bone marrow has been found suitable as matrix for EtG determination. The highest EtG concentrations have been found in urine in all but one case, where the resorption of ethanol had been incomplete. Second highest EtG concentrations have been detected in liver samples. In two cases with putrefaction, EtG could not be detected. In these cases, the detectable ethanol might have been produced partially or in total by postmortem fermentation. However, instability of EtG during putrefaction cannot be totally excluded which might result in a total loss of EtG.     

单细胞凝胶电泳分析死后小鼠骨骼肌细胞核DNA降解

目的研究小鼠死后骨骼肌细胞核DNA降解与死亡时间的关系。方法应用单细胞凝胶电泳(SCGE)技术结合荧光显微镜和专业的计算机图像分析技术,测定111只小鼠在死后72h内不同时间点骨骼肌细胞核DNA电泳形状的8项参数值,所得数据进行统计学分析。结果在个体死亡72h内,“彗星”的尾DNA含量比例、尾长、尾矩、O live矩、尾面积数值随时间的延长而增大;而“彗星”的头半径,头DNA含量比例和头面积数值则随时间的延长而减小。经对每个参数的测量值进行多项式运算,获得DNA降解趋势的二项式回归方程(P<0.001)和多元回归方程(P<0.0001),且均具有极显著性意义。结论在死后一定时间内,小鼠骨骼肌组织核DNA降解随死后经过时间的变化而呈一定规律性变化,DNA降解与死亡时间之间呈线性相关。     

利用尸体软组织生物力学性状时序性变化推断死亡时间

目的探讨尸体软组织生物力学性状时序性变化规律及其推断死亡时间的可行性。方法将实验大鼠用颈椎脱臼法处死,放置25℃恒温培养箱内即刻、6、12、18、24、30、36、42、48、60、72、96、120、144、168、192h,定时、定位、定向、定形切取大鼠胸部正中皮肤、腹壁肌肉、胸主动脉、结肠和小肠的生物力学试件,用电子万能材料试验机检测各试件的极限载荷、应变、应力生物力学参数。结果除主动脉外,尸体皮肤、肌肉、小肠、结肠的各项软组织生物力学参数均呈明显的死后时序性下降趋势。各参数下降趋势具有显著性差异的＂窗口期＂分别为：极限载荷：死后18h内皮肤、小肠呈下降趋势;24h内肌肉、结肠呈下降趋势;96～168h肌肉呈下降趋势;72～192h结肠呈下降趋势。应变：各组织在24h内均呈下降趋势,其结肠72～192h缓慢下降趋势。应力：皮肤18h内呈下降趋势,肌肉、小肠24h内呈下降趋势,结肠36h内呈下降趋势。结论大鼠尸体软组织各生物力学性状在不同的＂窗口期＂存在时序性变化规律,综合各指标可用于推断相应＂窗口期＂内的死亡时间。