家兔玻璃体液HBDH、LDH活性变化与PMI相关性的研究

目的 寻找一种推断PMI的新方法。方法 应用全自动生化检测仪(用比色法)检测家兔死后0-54小时玻璃体液内羟丁酸脱氢酶(HBDH)、乳酸脱氢酶(LDH)在两组温度下(25-30℃:10-15℃)的失活情况。结果 两种酶的活性在死后一定时间内各出现一个平台期,平台期后两酶的活性迅速下降,低温组在死亡54小时后活性几乎为零,高温组在死亡48小时后活性几乎为零。经统计学分析得知死后两种酶活性与PMI之间有显著的负相关性(P<0.05)。结论 在0-54小时内依据其各自的回归方程将可以大致地推断PMI,依据两种酶活性的多元回归方程可以较准确地推断PMI。     

死后大鼠骨骼肌运动终板和肌纤维酶,免疫组化研究

为进一步了解骨骼肌组织的死后变化特点和规律及其与死后时间（ＰＭＩ）的关系。本文选用成年实验大鼠,系列观察了死后０－９６小时骨骼肌运动终板和肌纤维在不同温度条例上ＡＣｈＥ（乙酰胆碱酯酶）,ＳＤＨ（琥珀酸脱氢酶）ＬＤＨ（乳酸脱氢酶）Ｃａ＾２＋－ＡＴＰａｓｅ（钙激活三磷酸腺苷酶）的酶组织化学变化及运动终板突触素的免疫组织化学变化,并使用图像处理系统对上述酶反应结果进行了定量分析和比较。结果发现,上述改变     

保存温度与时间对生物样品中氯胺酮稳定性的影响

目的研究不同温度和时间条件保存时生物样品中氯胺酮的稳定性。方法家兔以氯胺酮150mg/kg灌胃,30min后处死,取其血、肝、肾、脑,分别在室温（18～24℃）和冷冻（-20℃）条件下保存,并用气相色谱-质谱法定性分析、气相色谱-氮磷检测器法测定不同时间各样品中氯胺酮含量。结果血、肝、肾、脑冷冻保存至第30天氯胺酮含量均降低（P〈0.05）;室温条件下各样品中氯胺酮含量自第5天起均升高（P〈0.05）。结论生物样品在冷冻条件下保存时氯胺酮稳定性较好,怀疑氯胺酮中毒或死亡的检材应冷冻保存,尽快检测。     

《法医学杂志》文章中可直接使用的缩略词

PMI（postmorteminterval，死亡时间） 5-HT（5-hydroxytryptamine，5-羟色胺）     

家兔死后眼玻璃体液消光度与死亡时间关系研究

目的 为寻找一种精确推定死亡时间的方法。方法 应用 754分光光度计,在 420nm波长下,检测了 48只家兔死后不同时间的眼玻璃体液消光度,探讨其与死亡时间的关系。结果 眼玻璃体液消光度（ X）与死亡时间（ Y）呈线性正相关（ r=0.983 27, P< 0.05）,在死后 0～ 72小时,眼玻璃体液消光度与死亡时间存在回归关系（ F=116.54, P< 0.05）,直线回归方程为： Y = 453.30X＋ 0.75 [Y为死亡时间 (PMI),单位为小时; X为眼玻璃体液消光度 ]。结论眼玻璃体液消光度的测定可作为推断死后 72小时内死亡时间的参考指标之一。     

大鼠死后肝细胞核DNA的组织化学定量研究

推断死亡时间是法医学尸体检验须解决的重要问题之一。本实验用显微分光光度计定量测定了不同环境温度下（１０℃、２０℃、３０℃）大鼠死后不同时间肝细胞核ＤＮＡ平均含量，发现１０℃、２０℃、３０℃下大鼠死后肝细胞核ＤＮＡ含量与死亡时间呈负线性相关，其相关系数分别为—０．９９０５、—０．９８６３、—０．９７０２，且随环境温度升高ＤＮＡ含量的下降速度明显加快。作者认为ＤＮＡ组织化学定量技术可望成为推断死亡时间的新方法。     

不同温度下大鼠皮肤5种RNA指标与PMI的相关性

目的探讨大鼠皮肤内β-肌动蛋白(β-actin)、甘油醛-3-磷酸脱氢酶(glyceraldehyde-3-phosphate dehydrogenase,GAPDH)、18S核糖体RNA(18S ribosomal RNA,18S rRNA)、5S核糖体RNA(5S ribosomal RNA,5S rRNA)以及微小RNA-203(microRNA-203,miR-203)这5种RNA指标在不同温度下的表达水平与死亡时间(PMI)的相关性。方法将18只SD大鼠随机分为3组,处死后分别置于4℃、15℃和35℃的环境中,于死后0~120h内11个时间点取大鼠腹部皮肤。抽提皮肤总RNA,利用实时荧光定量PCR技术检测5种RNA表达水平。运用geNorm软件选取合适内参后,利用GraphPad软件对内参标准化的RNA指标进行回归分析。结果 5S rRNA和miR-203作为内参最合适。在4℃和15℃温度组中,β-actin和GAPDH的表达量变化与PMI线性关系良好。在35℃下,β-actin和GAPDH与PMI呈现S形曲线关系。而18S rRNA只在15℃和35℃温度组呈现一定的线性关系。结论皮肤组织比较适合作为提取RNA的检材,其中β-actin和GAPDH表达水平与PMI相关性良好,有望成为推测PMI的辅助指标。     

多温度下死后心血pH值随时间变化的三维拟合函数研究

目的 探讨通过测量多温度条件下家兔右心血液pH值进行死亡时间推断的可行性.方法 48只家兔随机分为6组,空气栓塞法处死后于10℃、15℃、20℃、25℃、30℃和35℃的不同死亡时间点采集右心血液;用PB-10型pH仪测量样本pH值;SPSS统计软件进行回归分析;MATLAB软件进行数学分析.结果 各温度组右心血液pH值变化与死亡时间呈高度相关(R2=0.974 ～0.982);获得拟合的曲面方程.结论 利用插值函数拟合pH值与死亡时间的曲面方程可进行温度变化条件下的死亡时间推断.     

北京地区尸生性蝇类研究及其在法医鉴定中的应用

为提供法医学尸体死亡时间鉴定的准确性，对北京地区人尸上蝇类区系进行了系统调查（1994～1996）。发现本地区共有双翅目尸生性蝇类3科、12属、14种。其中常见种有红头丽蝇、丝光绿蝇、大头金蝇、肥须亚麻蝇、急钓亚麻蝇、棕尾别麻蝇和家蝇7种。对其常见种的幼虫进行不同温度和湿度下培养，观察常见环境因素，如日历期、风雨天气等因素对其生长发育速率的影响。结果表明，蝇类幼虫在其适宜生长发育的温度范围内，随着温度的升高发育速率加快，历期缩短；在同一温度和同一饲养条件下，发育速率因种而异；平均温度在28℃左右时，蝇类幼虫发育最快，尤其是1～2龄幼虫，时间为1天左右，且较稳定。这些数据对法医学推断死亡时间具有参考价值。     

尸温与死后时间的研究进展

利用尸温推测死亡时间,是最常用的方法之一。但是,由于尸温的下降受到尸体内外诸多因素的影响,推测出来的死亡时间误差过大,影响其单独应用。但与其它方法相比较,它仍有很大的优越性,如仅靠温度数据就能作出死亡时间的推测。为了使其优越性充分发挥作用,国内外不少学者对尸温的下降规律进行了大量研究,将误差缩短到2小时以内,有的甚至缩短到1小时之内。为此,本文就尸温的研究与应用进展扼要综述如下: 1 尸温下降的原理 生活状态的人体,由于产热与散热的动态平衡,维持着机体温度相对恒定在一定的范围内,如腋窝温度在36.0～37.4℃;口腔温度在36.7～37.7℃;直肠温度在36.9～37.9℃。     

Time- and Temperature-Dependent Changes in Cytochrome c Oxidase Activity and Cyanide Concentration in Excised Mice Organs and Mice Cadavers

Postmortem stability of cyanide biomarkers is often disputed. We assessed the time and temperature-dependent changes in cytochrome c oxidase (CCO) activity and cyanide concentration in various organs of mice succumbing to cyanide. Immediately after death, excised mice organs and mice cadavers were stored at room temperature (35°C ± 5°C) or in frozen storage (−20°C ± 2°C). At various times after death, CCO activity and cyanide concentrations were measured in excised mice organs or organs removed from mice cadavers. The study revealed that (i) measuring both the biomarkers in mice cadavers was more reliable compared to excised mice organs, (ii) measuring temporal CCO activity and cyanide concentration in vital organs from mice cadavers (room temperature) was reliable up to 24 h, and (iii) CCO activity in the brain and lungs and cyanide concentration in organs from mice cadavers (frozen) were measurable beyond 21 days. This study will be helpful in postmortem determination of cyanide poisoning.     

Post-mortem changes in skeletal muscle protease and creatine phosphokinase activity — A possible marker for determination of time of death

The pattern of change in activity of two enzymes in rat skeletal muscle during body storage after death has been determined. Myofibrillar protease activity was found to increase linearly with time of storage post mortem at room temperature but not at 4 °C. In contrast, creatine phosphokinase activity declines linearly with time, and again storage at 4 °C prevented the change in enzyme activity. Starvation of animals for 5 days or forced exercise prior to death did not markedly alter the rate of change in activity of the two enzymes, although creatine phosphokinase specific activity at time of death was higher in the starved and exercised rats as compared to control animals. A plot of the logarithm of protease/creatine phosphokinase specific activities ratio versus time post mortem yields a linear curve at room temperature. These observations offer a potential method for estimating time of death.     

Viability of Human Articular Chondrocytes Harvested Postmortem: Changes with Time and Temperature of In Vitro Culture Conditions

Different studies of long‐term chondrocytes viability have shown a gradual reduction as a function of time and ambient temperature. The aim of our in vitro study was to establish chondrocyte postmortem viability curves for 4°C, 11°C, 23°C, 35°C during 63 days after the donors' death. Osteochondral cylinders were procured from the knees of 16 male donors (20–47 years), stored in preservation media that was not changed, and analyzed in 3‐day intervals using a confocal laser scanning microscope. A significant influence of time on viability was found from Day 9 (p = 0.0029) and onwards (p < 0.0001). The lowest overall chondrocyte viability was at 35°C, followed by 4°C (p < 0.0001). The conditions used in this in vitro analysis suggest that similar viabilities may occur while in situ in the decedent. Further studies of chondrocyte viability from individuals with known postmortem intervals may show premise to help evaluate time since death in the late postmortem interval.     

应用酶组织化学方法鉴定死亡时间的初步探讨

利用7例颅脑外伤死亡的健康青年尸体,在死后48h,环境温度18～24℃,空气相对湿度83～92％和实验湿度54～64％的条件下,检测肝脏、肾脏酶活性的变化。实验结果表明,肝脏乳酸脱氢酶(LDH)和L-苹果酸脱氢酶(MDH),随着死亡时间的延长,活性逐渐减低,48h近于阴性;而肾脏上述二种酶活性则在死亡后6h和24h出现高峰,36h开始下降;肝脏的酸性磷酸酶(ACP)亦于死后6h和24h出现高峰,36h开始下降。而肾脏此种酶在死后18～24h,有增高趋势。笔者认为上述酶活性的规律性变化有助于死亡时间的推断。应用二种以上酶活性的变化特点,能够较准确地判断死亡时间。     

The Optimal Combination of Cartilage Source and Apparatus for Long‐Term In Vitro Chondrocyte Viability Analysis

Abstract: Most studies of long‐term chondrocytes survival were for tissue banks. They showed a gradual reduction in the viable chondrocytes percentage as a function of time and ambient temperature, but the samples were harvested under optimal conditions. The aim of our study was to determine the most reliable combination of cartilage source and assay for the in vitro postmortem chondrocyte viability analysis in the conditions that imitate a dead body. Osteochondral cylinders were procured from femoral condyles and talar trochleas of three male donors and stored in the cell culture media at 4 ± 2°C and 23 ± 2°C. The samples were analyzed by a cell viability analyzer and a confocal laser scanning microscope (CLSM) initially 24–36 h after death and then in 4‐week intervals. The results reconfirmed the significant influence of time (p = 0.0002), but not of the temperature (p = 0.237). The largest reproducibility was presented for the knee joint and the CLSM.     

The Role of Vitreous Magnesium Quantification in Estimating the Postmortem Interval

The use of magnesium as a parameter to estimate the time of death is controversial. In 32 traumatic deaths with known postmortem intervals (PMIs), small quantities of vitreous humor (VH) were sampled repetitively every 3 h until 24 h postmortem. The bodies were kept at the constant ambient temperature of 20°C (68°F). The concentrations of magnesium were in the range of 0.47–1.15 mM. A statistically significant correlation of the concentration of magnesium with the PMI was observed (r = 0.453, p < 0.01), but with small predictive value—coefficient of variation for regression was 45.5%; the average of the paired differences between the estimated and actual PMIs was 5.24 + 3.19 h. Although useful results might be expected due to the large transmembrane gradient for magnesium, the results of this study strongly disprove the usefulness of measuring magnesium in VH to estimate the time of death.     

Autolytic changes in blood cells and other tissue cells of human cadavers. I. Viability and ion studies

The viability of white blood cells, spermatozoa of the epididymis, cells from minced spleen, lymph node and lung, and cells aspirated from the bone marrow of the sternum was studied using the vital dye exclusion test. The material comprised 123 medicolegal autopsy cadavers which had been stored in a mortuary cold room at +4 °C up to 10 days after death. Cells which excluded trypan blue were found in various specimens from all cadavers. However, there was marked individual variation in the results. The loss of viability of the white blood cells showed a moderate correlation (r = ?0.78) with the increase in the post-mortem (p.m.) time, whereas the results for other tissues were not so significant. The K⁺ and Mg²⁺ and haemoglobin content and the osmotic resistance of the red cells correlated poorly with the p.m. time. The present results suggest that despite the general assumption that autolytic changes proceed rapidly at the cellular level, individual cells and especially blood cells may remain viable for a long time in cadavers kept at +4 °C.     

损伤皮肤酯酶显微定位、定量与损伤时间推断

作者用显微分光光度计扫描法,对32只 Hartly 豚鼠的实验性损伤以及8例人体损伤的皮肤组织中的酯酶,进行定位、定量研究。发现生前损伤后,创壁0.4mm 以内真皮胶原纤维等间质即刻出现酯酶着色,存活时间越长,着色就越深、范围越大。生前各时间组互相间有显著差异。所有的生前损伤组创缘酯酶含量及平均单位面积酶含量与正常皮肤及死后损伤组差别均极显著。人体损伤皮肤组织酯酶定位及含量变化与豚鼠所见一致。因此,作者认为损伤皮肤酯酶定位定量分析以推断损伤后存活时间,已可应用于法医实际案例鉴定中。作者还根据对抑制剂的反应认为创缘真皮胶原纤维上所显示的酯酶为 B-酯酶。     

大鼠死后骨骼肌细胞核DNA降解的彗星电泳检测

目的探讨彗星电泳检测大鼠死后骨骼肌细胞核DNA降解碎片与死后时间的关系。方法建立SD大鼠死亡模型,18只雌性大鼠断颈处死,在25.1℃分别于死后6、12、24、36、48、60提取大鼠骨骼肌组织进行彗星电泳,荧光显微成像系统采集图像,同时获取多个分析参数(Comet4.0),并进行统计学处理。结果在死后6～48h,彗星尾长(TL)在死后各时间点的均数依次为11.56±0.10、17.76±0.18、18.82±0.21、21.68±0.18和23.33±0.07;Oliver尾矩(TM)在各时间点的均数依次为1.63±0.46;2.12±0.90;2.15±1.03;2.22±0.76;3.35±0.80;尾DNA(TDNA)在各时间点的均数依次为29.57±8.42;32.36±10.92;30.11±12.55;37.81±12.03;54.76±8.60。结论细胞DNA降解随着死后时间的延长而增加;彗星尾长用于推断死后时间优于Oliver尾矩和尾DNA。     

Developmental Times of Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) at Constant Temperatures and Applications in Forensic Entomology

The characteristic life stages of infesting blowflies (Calliphoridae) such as Chrysomya megacephala (Fabricius) are powerful evidence for estimating the death time of a corpse, but an established reference of developmental times for local blowfly species is required. We determined the developmental rates of C. megacephala from southwest China at seven constant temperatures (16–34°C). Isomegalen and isomorphen diagrams were constructed based on the larval length and time for each developmental event (first ecdysis, second ecdysis, wandering, pupariation, and eclosion), at each temperature. A thermal summation model was constructed by estimating the developmental threshold temperature D₀ and the thermal summation constant K. The thermal summation model indicated that, for complete development from egg hatching to eclosion, D₀ = 9.07 ± 0.54°C and K = 3991.07 ± 187.26 h °C. This reference can increase the accuracy of estimations of postmortem intervals in China by predicting the growth of C. megacephala.