microRNA在急性心肌梗死法医学诊断中的应用前景

急性心肌梗死(acute myocardial infarction,AMI)的诊断一直是法医病理研究的重点和难点。在心肌缺血早期,组织病理学检查很少出现具有诊断价值的病理改变。近年来,利用分子指标进行AMI早期死后诊断成为法医研究热点之一。其中,microRNA在临床上已经被广泛研究可用于AMI病人的诊断,并且具有极高的敏感性和特异性。基于此,本文综述了临床病人样本和动物实验中利用microRNA诊断AMI的研究现状,并重点对microRNA死后稳定性进行总结,以期探讨microRNA在AMI法医学诊断中的应用前景。     

Utilization of serum tryptase and immunoglobulin e assay in the postmortem diagnosis of anaphylaxis

The postmortem diagnosis of anaphylaxis is difficult. Serum concentrations of tryptase (a mast cell product released during anaphylaxis) have been used after death as an indicator of possible antemortem anaphylaxis. However, studies have indicated that tryptase may be elevated with increasing postmortem interval (PMI), or in nonanaphylactic deaths with significant atherosclerosis or chest trauma. Serum total IgE has been used by some to confirm anaphylaxis when tryptase is elevated. Serum levels of tryptase from 57 decedents with varying PMI, all dying of presumed nonanaphylactic causes, were determined. In cases with elevated levels (>11.4 ng/mL), an assay of total serum IgE was also performed. Both tryptase and IgE demonstrated significant elevations with increasing PMI. Decedents were categorized according to presence of cardiovascular disease, chest trauma, or both; many demonstrated elevation of 1 or both markers, without statistically significant differences between categories. Postulated mechanisms for nonanaphylactic elevations of these markers are reviewed. The possible utility of allergen-specific IgE or allergen panels is discussed.     

3种特殊染色在急性心肌梗死死后诊断中的应用比较

目的对比观察苏木素-碱性复红-苦味酸染色(HBFP染色)、变色酸2R-亮绿染色和Heidenhain染色在急性心肌梗死死后诊断中的应用价值。方法以大鼠急性心肌梗死模型、法医检案急性心肌梗死心脏标本作为研究对象,采用HBFP染色、变色酸2R-亮绿染色和Heidenhain染色进行对比观察。结果①3种特染方法在大鼠心肌缺血15min时均可观察到阳性染色,且阳性染色面积随缺血时间的延长而扩大;②大鼠急性心肌缺血4h心脏标本在-20℃、4℃及室温条件下保存至14d,3种特染方法仍可见阳性着色,但变色酸2R-亮绿染色和HBFP染色随保存时间的延长而出现着色能力下降,阳性区域变小的趋势,Heidenhain染色效果最为稳定;③急性心肌梗死检案标本中,3种特染方法均可显示缺血心肌纤维,发病1h内死亡者Heidenhain染色优于另外两种染色。结论 3种特染均可客观的显示出急性心肌梗死早期病理改变,其中Heidenhain染色更具稳定性和可操作性。     

The use of rapid diagnostic test of procalcitonin serum levels for the postmortem diagnosis of sepsis

Because serum Procalcitonin is reported to be a valid postmortem marker of sepsis, this prospective study was carried out to determine whether the semi-quantitative PCT-Q((R))-Test (B.R.A.H.M.S., Germany) is a reliable indicator of postmortem Procalcitonin (PCT) serum levels, thus enabling a quick "tableside" diagnosis of sepsis. Postmortem PCT-levels of 70 forensic and 78 clinical-pathological autopsy cases (n=148) were examined using the B.R.A.H.M.S-PCT-Q-Test during autopsy. 27 cases were categorized as the cases of sepsis according to the ACCP/SCCM Consensus Conference criteria. 121 cases were assigned to the non-sepsis group. Among the 148 cases, 18 samples could not be analyzed by the reason of strong hemolysis. Using a cut-off point of 2 ng/ml, 20 cases of sepsis were identified (true positive) whereas 3 cases of sepsis were not detected (false negative). In the non-sepsis group (107 cases) 6 cases showed a positive testing (false positive). When applied within 48 h postmortem, the PCT-Q-Test showed a sensitivity of 86.96% and a specificity of 94.39% (at cut-off 2 ng/ml). Likelihood ratios and positive predictive values proved to be lower in the forensic autopsy group (PPV: 59.3% in forensic case vs. 85.1% in clinicopathological cases; NPV: 98.73% in forensic cases vs. 95.2% in clinicopathological cases). The PPVs using a cut-off point of 10 ng/ml were 100% in both groups independent of sepsis prevalences. The results show, that a high NPV for prevalences ranging from 3% to 30% can be reached using a 2 ng/ml cut-off point, whereas a cut-off of 10 ng/ml ensures a high PPV for the respective prevalences in the absence of exclusion criteria. The study provides strong evidence that the introduction of rapid diagnostic test (RDTs) of postmortem PCT serum levels may be useful in achieving rapid distinction between sepsis and non-sepsis-related causes of death, especially in conjunction with the medical case history and further autopsy results. In addition, the use of RDTs enables clinicians to conduct an evidence-based validation of clinical diagnosis, thus facilitating future clinical decision-making.     

On the possibility of postmortem diagnosis of diabetes mellitus

The present study has demonstrated that elevated level of glycated haemoglobin in cadaveric blood may be regarded as an absolute diagnostic criterion for diabetes mellitus irrespective of other macro- and/or micropathomorphological signs of this disease.     

Evaluation of histologic changes of the skin in postmortem period

ABSTRACT: Determination of the time of death is an important consideration in forensic practice. Many methods have been attempted to accurately and systematically determine the postmortem interval (PMI). Histologic examination of the skin or appendages is one of the methods tried by few researchers. However, no attempt had been made to analyze the histologic changes in the skin and appendages simultaneously and to compare them with PMI. We sequentially studied the histologic changes of the skin and appendages in the early PMI. The results of the present study show that the skin undergoes progressive morphological changes in the postmortem period. The epidermis and the dermis appeared normal for 6 hours after death, and after this period, degenerative changes began. By 6 to 9 hours after death, degeneration began in the dermis, and by the end of 18 hours, the dermis began to disintegrate. The sweat glands appeared normal for approximately 3 to 4 hours. For 18 hours after death, the sebaceous glands and hair follicles appeared normal, and after this period, degeneration began.     

类胰蛋白酶和IgE测定在过敏性猝死诊断中的应用研究

目的探讨过敏性猝死法医学鉴定的诊断方法和指标。方法采取10例正常人、9例过敏性猝死和19例其他死因（排除过敏反应、冠心病）尸体的静脉血，采用荧光酶联免疫法（Pharmacia UniCAP100过敏原定量分析仪）和酶联免疫吸附试验ELISA法分别测定血清肥大细胞类胰蛋白酶和]gE含量，采用免疫组化方法观察过敏性猝死和其他死因的肺组织中的肥大细胞类胰蛋白酶免疫组化染色。结果过敏性猝死者的血清类胰蛋白酶和IgE含量升高，与其他死因之间的差异具有显著性意义（P〈0．01），其他死因和正常人之间的差异无统计学意义（P〉0．05）；与其它死因相比，过敏性猝死肺组织中的肥大细胞类胰蛋白酶免疫组化阳性染色增强（P〈0．01）。结论过敏性猝死者血清IgE和肥大细胞类胰蛋白酶含量显著升高；过敏性猝死者肺组织中肥大细胞类胰蛋白酶染色增强。     

Fatal MDMA-intoxication with postmortem diagnosis of hyperthermia

Hyperthermia is one of the main symptoms of the toxic MDMA effect. In a fatal intoxication, the premortal increase in body temperature may lead to a false assessment of the time of death. The problems of drug-induced hyperthermia faced by forensic pathologists and criminal investigators are discussed on the basis of a death from the autopsy material of the Berlin Institute of Legal Medicine (CCM).     

Vitreous humor carbohydrate-deficient transferrin concentrations in the postmortem diagnosis of alcoholism

Deaths from the effects of alcohol intoxication are encountered routinely in forensic practice. In an important number of cases difficulty may arise in interpreting the significance of results obtained in the autopsy. In clinical practice biochemical markers, particularly serum gamma-glutamyl-transpeptidase (GGT), alanine aminotransferase (ALT), aspartate transaminase (AST), carbohydrate-deficient transferrin (CDT), and erythrocyte mean corpuscular volume are used to diagnose heavy alcohol consumption. CDT is used as a reliable and specific marker. In postmortem diagnosis, because of the difficulty in interpreting blood alcohol levels and relatively non-specific pathological features, biochemical compounds have been studied for use as possible markers. The aim of this study was to evaluate the usefulness of the postmortem determination of CDT in vitreous humor as a confirmation of antemortem alcoholism. CDT levels were studied in 66 male cadavers with a mean age of 55.9 years (S.D. 17.0, range 22-87 years) with a mean postmortem interval of 17.9 h (S.D. 11.4, range 4-72 h). Cases were assigned to two diagnostic groups according to the antemortem diagnosis of alcoholism. Statistically significant differences were found for CDT and ALT concentrations between the two diagnostic groups. The highest vitreous humor levels of CDT and ALT were obtained in the group of cases with a previous diagnosis of alcoholism. Our results suggest that vitreous humor CDT levels are useful in cases where the postmortem diagnosis of alcoholism is hindered by the non-specificity of data.     

Site of discovery in the bath tub. Evaluation of a fatality after four years using postmortem roentgen diagnosis]

On the one hand the following case demonstrates the problems of finding a dead body in the bathroom and on the other hand the importance of postmortem x-ray examinations. The corpse of an 65-year-old woman was discovered in her bath tub. At the body's feet there was a hair dryer. Investigations concerning murder were negative, circumstances appeared to be unsuspicious. That was why a postmortem examination was not performed. -4 years later murder had to be considered. After manual strangling the deceased and the hair dryer were layed into the bath tub. The corpse was exhumed and taken to a forensic autopsy. It was found that the hyoid bone and the superior horn of the thyroid cartilage were both fractured on the right. Damages caused by the preparation itself were excluded by using serial roentgenographs. These observations in combination with histological findings and the results of further investigations convincingly supported the suspicion of strangling by hand.     

Differential diagnosis of vital and postmortem processes in burn cadavers

After presenting the most important intravitam reactions in 114 burnt corpses of the G?ttingen autopsy material covering the years 1969-83 two cases are described with a) a so called burn haemorrhage in the stomach b) an extremely wide spreading of the upper thighs. Differential diagnosis and forensic significance of these findings according to chemical, histopathological and experimental examinations are discussed.     

Comparison of different techniques for the postmortem diagnosis of myocardial infarction

A study was performed on 74 medicolegal autopsy cases for the purpose of comparing the reliability of four different techniques (haematoxylin-eosin stain, acridine orange method, formazan test and K/Na ratio) used for the postmortem diagnosis of myocardial infarction. Although the results showed that the formazan test and K/Na ratio give a good reliability to rule out a myocardial infarction, the most specific technique for the diagnosis of infarct is the formazan test.     

纤维连接蛋白诊断心肌梗死的死后稳定性研究

建立狗急性心肌缺血模型,应用免疫组织化学和图像分析技术,对用纤维连接蛋白诊断心肌梗死的稳定性进行研究。心肌缺血组织随死后放置时间延长,纤维连接蛋白阳性反应面积逐渐减少,但放置4周仍呈阳性反应;而正常心肌组织放置不同时间,均未见阳性反应。纤维连接蛋白在心肌梗死死后诊断上具有很好的稳定性,在法医学实践中具有重要实用价值。     

Evaluation of postmortem redistribution phenomena for commonly encountered drugs

We described the findings of a study into the post-mortem redistribution (PMR) of 76 drugs found in 129 drug-related cases between 2006 and 2009. Seventy six drugs (psychotropic drugs (n=14), antidepressants (n=9), sedatives (n=6) and so on) were simultaneously quantified in cardiac and peripheral blood by gas chromatography-mass spectrometry (GC/MS) or liquid chromatography-tandem mass spectrometry (LC/MS/MS). The absence, possibility or presence of PMR of drugs was determined according to the ratios of cardiac to femoral blood concentrations (C/P ratios). Proxyphylline (C/P ratio: 0.85) showed no PMR; carbamazepine was not subject to PMR; a potential for PMR of lorazepam and mirtrazapine cannot be excluded; chlordiazepoxide is subject to PMR; acetaminophen and alprazolam exhibit minimal PMR; amitriptyline and benztropine exhibit PMR. Codeine (C/P ratio: 4.9), zolpidem (C/P ratio: 3.74), chlorpromazine (C/P ratio: 2.97), fluoxetine (C/P ratio: 2.83) and propranolol (C/P ratio: 2.72) had the largest C/P ratios. Postmortem drug concentrations showed variations depending on sampling sites and characteristics of the drugs. It is continuously necessary to analyze commonly used or abused drugs in simultaneously collected cardiac and peripheral blood to establish significant reference values for PMR. These findings can be used to reach a conclusion about the cause and manner of death.     

早期心肌梗死死后诊断的法医病理学研究进展

心源性猝死在成人猝死的死因中占首位,心肌梗死是冠心病猝死的主要机制之一,由于早期心肌梗死发生突然,且可在短时间内死亡,肉眼和镜下都缺乏典型的形态学改变,一直都是法医实践中研究的热点,本文回顾多年来心肌梗死的研究成果在法医学中的应用及局限,展望新的临床指标在法医工作中的可行性,期望提高死后心肌梗死诊断的准确率。     

RNA指标在死后人体皮肤组织中稳定性研究

目的探讨多种RNA指标的表达水平在死后人体皮肤组织中的稳定性。方法收集死亡时间(PMI)明确的人体皮肤组织样本10例,提取总RNA,运用实时荧光定量PCR(qRT-PCR)技术检测ACTB、GAPDH、18S、miR-203、5S、1et-7a、RPS29、U6共计8个RNA指标的表达水平。geNorm软件评估各RNA指标表达水平的稳定性,挑选出稳定的内参指标并分析其表达水平与相关因素(年龄、性别、死因)的关系。结果人体皮肤组织中5S、U6和miR-203表达水平最为稳定,其平均表达量与年龄、性别、及死因无关(P0.05)。结论 5S、U6和miR-203可以作为qRT-PCR方法研究PMI的理想内参指标。     

Ethanol-oxidizing enzymes of the adrenal glands in acute alcoholic intoxication

Complex morphological-and-histological examinations of the adrenal glands were performed on actual forensic-medical samples in acute alcoholic intoxication (AAI), i.e. 107 death cases of men and women, aged 17 to 60, due to mechanical trauma, lethal ethanol intoxication and somatic complications of chronic alcoholism. A differentiated distribution of ethanol-oxidizing enzymes was defined, i.e. of alcohol-dehydrogenase and acetyl-dehydrogenase in the structural-and-functional zones of the adrenal cortex and medullary substance. A differential activity of enzymes was established that correlated with the level of exogenous alcoholemia. Sets of morphological signs related with examinations of the adrenal glands, which enabled a differential diagnosis of death causes in AAI, were specified.     

Comparative evaluation of postmortem serum concentrations of neopterin and C-reactive protein

The cellular immune response is accompanied by the release of neopterin. The level of neopterin in serum is increased in patients suffering from viral infections, autoimmune diseases, systemic inflammation, allograft rejection and malignant diseases, while that of C-reactive protein (CRP) is known to rise during inflammatory diseases and traumas. To investigate postmortem neopterin and CRP concentrations with regard to the cause of death, we examined cardiac and peripheral blood samples in 474 autopsy cases without advanced decomposition (0-96 years of age, 343 males and 131 females), 2.8 h to 3 days (median, 18.0 h) after death. Survival time was 0.1 h to 5 months (median, 3.0 h) for traumatic death, and 0.1-1, 440 h (median, 2.5 h) for natural death. In autopsied subjects, neopterin concentrations were higher than the clinical reference, independent of the time after death, and depended on the survival time. In cases of acute and subacute death due to trauma, the neopterin level in right heart blood was mildly to moderately elevated (about 50-200 nmol/l) except for sharp instrument injury, whereas the CRP concentration usually remained low (<1 mg/dl). However, a moderate rise in the CRP level (around 1-10 mg/dl) was observed in fatal cases of hypothermia (cold exposure). Markedly elevated serum CRP and neopterin levels (>10 mg/dl and >500 nmol/l, respectively) were detected in cases of delayed death due to trauma involving systemic inflammatory response syndrome (SIRS) and of fatal bacterial infections. For sepsis, the serum CRP level was markedly elevated but the neopterin level was low in some cases. Fatal viral infections usually resulted in a marked elevation in the serum neopterin level (>500 nmol/l) with a mild to moderate rise in the CRP level. Combined analyses of neopterin and CRP may be useful to investigate viral infections and delayed traumatic death involving SIRS to support pathological findings.     

心肌梗死6项免疫组化指标的死后稳定性比较

目的　探讨用于早期心肌梗死死后诊断的维连接蛋白 (Fn)、纤维蛋白原 (Fg)、补体 (C5 )、肌红蛋白(Mb)、肌动蛋白 (HHF3 5 )、结蛋白 (Dm )等 6项免疫组化指标在死后不同时间的稳定性。方法　应用免疫组织化学法、图像分析和统计学处理系统 ,检测缺血心肌细胞内Mb、HHF3 5、Dm的缺失面积和Fn、Fg、C5的阳性反应面积 ,并对 6项免疫组化指标在死后不同时间的稳定性进行比较。结果　正常心肌组织 4℃放置 1～ 2d ,Dm、HHF3 5、Mb染色均匀 ,未见明显缺失 ;放置 3d以上 ,即可见Dm、HHF3 5、Mb明显缺失 ,且随放置时间延长 ,缺失面积逐步增大。缺血心肌组织随放置时间延长 ,Dm、HHF3 5、Mb的缺失面积逐步增大 ,Fg、C5、Fn阳性反应面积逐渐减少 ;放置 14d以上 ,Fg呈阴性反应 ;放置 2 1d以上 ,C5呈阴性反应 ;放置 2 8d ,Fn仍呈阳性反应。正常心肌组织放置不同时间 ,均未见Fg、C5、Fn阳性反应。图像分析结果显示阳性反应面积逐步减少。结论　Dm、HHF3 5、Mb的稳定性最差 ,易受死后自溶的影响 ,只适用于新鲜尸体 (死后 1～ 2d) ;Fg次之 ,可用于死后 4℃放置 7d的尸体 ;C5较好 ,可用于死后 4℃放置 14d的尸体 ;Fn稳定性最好 ,可用于死后 4℃放置 2 8d的尸体。     

Usefulness of different myocardial sampling zones for the postmortem diagnosis of myocardial infarction

The diagnosis of myocardial infarction requires the use of a group of tests that are very efficient, quick and inexpensive. Another important consideration is the choice of myocardial sampling zones, especially in cases of differential diagnosis between a cardiac injury secondary to a trauma or violent asphyxia and others, secondary to myocardial infarction. The aim of this work was to choose, through discriminant analysis, the most useful zones of cardiac tissue for the quantification of free fatty acids and free carnitine and for the performance of the K/Na quotient, as biochemical parameters for the postmortem diagnosis of myocardial infarction. According to the discriminant analysis performed, seven zones of cardiac tissue are necessary to achieve a differential diagnosis among "myocardial infarction," "other natural deaths," and "violent deaths" with a 71.9% efficacy. Greater diagnostic efficacy was found (78.1%) for differentiating between "natural deaths" and "violent deaths."