Localization and quantification of histamine in injured skin as parameters for the timing of wounds.

Localization and estimation of the histamine (HA) content in skin wound edges in 86 Sprague-Dawley rats and three cases of human injuries were carried out by a microfluorimetric method specific for this amine which forms a complex with o-phthalaldehyde (OPT). Distribution and density of the mast cells in the same areas were observed at the same time by toluidine blue stain. In all skin specimens with antemortem wounds, both the epidermis and upper dermis exhibit extracellular yellowish fluorescence of the HA-OPT complex. The fluorescent zone spreads in the wound edges with the lapse of time in vital injuries. The HA content increases gradually up to 30 min and then the yellow histamine fluorescence in areas 0-200 microns from the wound edge decreases. None of these features can be observed in normal skin and postmortem-injured skin. Mast cell degranulation can be demonstrated in all antemortem-injured skin. No statistical relationship exists between the number of mast cells and the HA-OPT fluorescence in either ante- or postmortem-injured groups. This study indicates that skin HA microfluorimetry by the OPT method is of practical value for distinguishing ante- from the postmortem wounds and for timing antemortem wounds.     

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。     

Localization and quantification of the nonspecific esterase in injured skin for timing of wounds.

Histochemical quantification of the nonspecific esterase (NSE) in injured skin was performed using histochemical demonstration of the enzyme and a microspectrophotometric scanning technique on specimens taken from 32 Hartley guinea-pigs and 8 cases of human skin wounds. In all antemortem incisions and lacerations, including those made at the agonal stage, NSE activity could be observed in the dermal tissue of the wound edge. The enzyme activity increases with the antemortem duration of the injuries. Both total content and mean concentration of NSE in the wound edge between antemortem and postmortem wound groups differ significantly (less than 0.01). Multiple range test shows that significant differences (P less than 0.05 or P less than 0.01) of total content of NSE in the wound edge also exist in 0-5 min, 15-30 min, 1-h, 2-h and 4-h antemortem incised wound groups and in 0-min, 5-min, 15-30 min, 1-h and 4-h antemortem laceration groups. The positive NSE reactions in 8 cases of human skin wounds were similar. The study indicates that histochemical quantification of NSE in injured skin is very useful in timing wounds and is exactly applicable in medicolegal practice. According to the different influences of inhibitors on enzymes, it was inferred that the enzyme activity in wound edges was due to B-esterase.     

损伤皮肤中组胺荧光显微定位定量与损伤时间推断的实验研究

本文应用显微荧光分光光度计法研究大白鼠皮肤创缘组织中组胺的分布和含量,并用甲苯胺蓝法观察创缘肥大细胞形态和数量的变化。结果发现,创缘真皮乳头层出现扩散的细胞外黄色荧光和肥大细胞脱颗粒现象为生前伤的重要特点;组胺荧光分布范围及增多的程度与损伤时间密切相关。从而为损伤时间推断提供了一种新的方法。     

对纤维蛋白诊断生前伤的评价——45例枪弹创组织的MSB染色和扫描电镜对比观察

生活反应是诊断生前伤的主要根据.近二十年来,法医病理学在研究生前伤和死后伤的诊断中,采用了多种新兴技术,如扫描电镜技术、酶组织化学技术、酶标技术、电泳技术等,促使生前伤的诊断水平有明显提高.伤后存活1小时死亡者,大部分皆可获得明确诊断;立即死亡或在濒死期形成的损伤,由于生活反应甚微,或受腐败等     

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。     

扫描电镜下缢沟的形态特征

作者用光镜和扫描电镜对縊沟进行对比观察,得出如下结论:(1)扫描电镜下縊沟组织的改变具有较恒定的特征,出血及纤维蛋白检出率远较光镜为高;(2)在光镜下生活反应不明显的生前缢沟,在扫描电镜下其表面及断面均可见红细胞及纤维蛋白形成,并包裹红细胞。縊沟部位胶原纤维之间可见散在大小不等的圆球形脂肪样小滴。缢沟部位肥大细胞的形态改变,有助于生前与死后缢沟的鉴别诊断。     

Complement component C3a or C3a desArg as a new marker for estimation of local vital reactions in incised skin wounds.

The anaphylatoxin C3a or its desArg form (C3a/desArg) generated during complement activation could be detected in the vicinity of incised skin wounds of guinea pigs using immunoblotting methods. The C3a/desArg peptides were detectable immediately after injury in local sites up to 3 mm from the wound edge. In subsequent determinations of up to at least 3-day-old antemortem wounds, the maximum concentration of these peptides was largely localized up to 6 mm from the wound edge at 2 h after injury. In postmortem wounds, however, these peptides were undetectable. When they were released in antemortem wounded tissues they could be detected up to 1 day at 22 degrees C after death. These results suggest that the detection of C3a/desArg in wounds using immunoblotting methods can be useful for distinguishing ante- from postmortem wounds.     

枪弹创纤维蛋白形成能力的免疫组化研究

采用免疫组化PAP染色法，对57例人体枪弹创标本的纤维蛋白形成能力进行检测。结果发现，所有生前枪弹创均有纤维蛋白形成祖创缘邻近皮肤组织内毛细血管内皮细胞以及血管内充盈的红细胞膜上亦呈棕色阳性染色；死后30min内形成的枪弹创均有纤维蛋白形成；死后37min后形成的枪弹创无纤维蛋白形成，说明死后30min，纤维蛋白形成能力仍存在。作者认为，仅根据损伤局部有纤维蛋白形成即判断为生前伤是不恰当的，尚应结合损伤部位、损伤程度、损伤类型以及其它生活反应综合评定，不宜单凭纤维蛋白判断。     

Frozen cadaver. Antemortem versus postmortem.

Three categories of skin from Hanford miniature swine were examined microscopically: normal, unfrozen skin; skin exposed to -75 degrees C air for 5 min, 6 h postinjury; and skin frozen postmortem. Frostbitten skin (antemortem freezing) was characterized grossly by a purple discoloration and microscopically by dilated, blood-filled, superficial capillaries. Other changes in the frostbitten skin were vacuolated epithelial cells and dermal edema. Unfrozen skin (controls) and skin frozen postmortem were more difficult to differentiate. However, the epidermis of the latter usually was compressed and more basophilic. We concluded that skin frozen antemortem could be separated from skin frozen postmortem by its intense hyperemia, characterized grossly as a purple discoloration and microscopically by engorged capillaries.     

死后肌肉切创创壁纤维蛋白的扫描电镜观察

用扫描电镜观察大鼠肌肉组织切创创壁纤维蛋白网的形成和形态,发现生前和死后8小时内的切创壁,均有纤维蛋白网形成,两者形态无显著差异。死后16小时,切创创壁无纤维蛋白网形成。     

Distinguishing antemortem from postmortem injuries by LTB4 quantification

Leukotriene B₄ (LTB₄) in skin samples from seven forensic cases was detected by HPLC to distinguish their antemortem or postmortem origin. In total, there were thirteen antemortem and seven postmortem specimens. The results showed that LTB₄ was found in all antemortem wound specimens which were either fresh, or refrigerated or fixed in formalin for less than 10 days. In contrast, LTB₄ could not be detected in postmortem wound specimens. These results suggested that detecting the content of LTB₄ is a useful method for distinguishing antemortem from postmortem injuries.     

Pseudo-Stab Wounds: Putrefactive Dehiscence of Remote Surgical Incisions Masquerading as Stab Wounds

Abstract:  Artifacts due to decomposition can be mistaken for antemortem injury leading to an incorrect suspicion of foul play. We describe an instance of postmortem wound dehiscence that mimicked antemortem stab wounds. A man with a history of colon cancer and substance abuse was found dead. There were advanced putrefactive changes and multiple defects of the anterior torso that resembled stab wounds. Subsequent investigation revealed that 11 months earlier, he had a laparoscopic-assisted colon resection that involved surgical incisions corresponding in location and size to the above defects. Putrefactive gases may cause bloating of the body. This distension may cause recent and remote healed incisions to dehisce. Correlation of these "defects" at autopsy with the antemortem clinical history is crucial in determining their etiology.     

Immunohistochemical detection of fibronectin and tenascin in incised human skin injuries

Immunohistochemical detection of molecules involved in inflammatory reaction can be useful for the diagnosis of vitality in skin wounds. We studied the expression of fibronectin (FN) and tenascin (TN) in 58 human skin wounds (48 vital and 10 postmortem). The age of vital injuries ranged from 3 min to 8 h and postmortem specimens were collected after a postinfliction interval of 15-180 min. One hundred thirty-seven formalin-fixed paraffin-embedded sections (mean: 2.3 sections per case) were stained with each of two monoclonal antibodies against FN and TN using the streptABC technique. A reticular staining for FN in wound edge and dermis was observed in 50% of vital specimens versus 0% in postmortem cases. Immunoreactivity was reduced in 10 autolysed cases. FN positivity exclusively at the injury margin was observed in 39.4% of vital wounds and 10% of postmortem cases. TN was negative in all specimens. Vital and postmortem hemorrhage areas showed positivity for FN and TN. Due to its low sensitivity, immunohistochemical analysis of FN is useful for determining vitality only in a minority of cases. Different factors in everyday practice, including autolysis and technical problems often produce false negative reactions with the result that FN cannot be regarded as a reliable parameter of vitality. Positive reactions (network staining) are more valuable than negativity but are not pathognomonic. Both vital and postmortem hemorrhages show an enhanced positivity for FN and TN, thus impeding the diagnosis.     

小鼠皮肤切创IL-10和IL-4的表达及其与损伤时间关系

目的　探讨IL 10、IL 4在小鼠皮肤切创愈合过程中的表达变化规律及其与损伤时间的关系。方法　应用免疫组织化学及图像分析技术 ,观察实验小鼠不同时程的生前伤 ( 0 5～ 168h)及死后伤 ( 1～ 6h)皮肤创伤局部IL 10、IL 4的表达情况。结果　生前伤 ,IL 10在伤后 1～ 3h阳性反应逐渐增强 ,3h达峰值 ,2 4h降至较低水平 ,伤后 48h再次升高 ,72h又达新峰值 ,其阳性表达部位主要在表皮细胞及单核 /巨噬细胞 ;IL 4于伤后 2 4h出现明显阳性反应 ,96h表达水平达峰值 ,168h仍维持在较高水平 ,其阳性细胞主要为创伤局部的成纤维细胞。死后伤 ,IL 10仅于死后 1～ 3h呈阳性染色 ,IL 4未见阳性染色。结论　IL 10、IL 4在小鼠皮肤切创愈合过程中的表达呈现一定的时序性变化。     

A preliminary study of skin electrical injury with computerized image analysis

This paper reports the application of computerized image analysis to a quantitative study of skin electrical injury in order to provide an objective and quantitative standard for identifying the electrical mark. The differences between antemortem and postmortem electrical injuries were also studied. In normal and electrically-injured human skin, the long/short axes ratio of nuclei were measured. The mean of epithelial basal cells was 1.4344 and 5.9325, respectively, the mean of hair follicle cells was 1.6193 and 4.5988, respectively, and the mean of capillary endothelial cells was 4.0229 and 4.3593, respectively. In rat specimens, the long/short axes ratio of nuclei of normal skin, at 15 and 5 min ante-mortem and at 5 min postmortem injury, the mean epithelial basal cells was 1.4124, 4.5417, 5.2282 and 5.0447, respectively. The mean of hair follicle cells was 1.4140, 4.4863, 4.1146 and 4.4522, respectively, while the mean of capillary endothelial cells was 2.8398, 3.7514, 3.6159 and 3.6977, respectively. The results indicate that the differences between electrical injuries and normal skin with respect to the ratios of long/short axes of the epithelial basal cell nuclei and hair follicle cell nuclei are of remarkable significance. On the other hand, no significant difference could be found between antemortem and postmortem injuries, nor between electrical injury and normal skin, concerning the long/short axes ratios of capillary endothelial cell nuclei. This result provides a preliminary objective standard for the changes of electrical skin injury.     

细胞因子(IL-6、TNF-α)在损伤时间中原位杂交法研究

采用原位杂交法，研究大鼠切削皮肤中细胞团子IL－6、TNF－αmRNA表达量，旨在探讨IL－6、TNF－α推断法医损伤时间的应用价值以及其在损伤生活反应中的分子机制。研究结果表明，根据大鼠损伤皮肤中TNF－αmRNA表达量能够区别生前伤与死后伤，并可以利用TNF－αmRNA表达量改变准确区别60min内和60min后损伤时间，但IL－6mRNA表达量在研究组内均未见阳性反应，不能推断损伤时间。     

Skeletal identification using the frontal sinus region: a retrospective study of 39 cases

The importance of identification using the frontal sinus has been previously demonstrated in case reports. In this study, 39 cases of identification using frontal sinus comparison from the Ontario Chief Coroner's Office were reviewed and differences between antemortem and postmortem radiographs examined. All cases involved decedents older than twenty years. Three cases were rejected due to poor antemortem and postmortem film quality. One subject had no frontal sinus. Thirty-five cases provided conclusive postmortem to antemortem pattern matches. Sixteen cases also yielded metric (quantitative) matches. Duration between antemortem and postmortem radiographic examinations, age, gender, and cause of death did not affect the ability to obtain a match. This is the largest study undertaken on actual cases and demonstrates the validity of frontal sinus pattern matching for forensic identification.     

大鼠皮肤切创后E-选择素表达及稳定性研究

目的探讨大鼠皮肤切创后E-选择素表达规律及法医学意义。方法健康SD大鼠90只,随机分成4组：正常对照组、活体切创组（30min～7d12个时间点）、死后切创组（30min～3h3个时间点）、死后稳定性组（-20％6h-7d9个时间点,25％6h～3d5个时间点）。在大鼠头部建立皮肤切创模型,按设定的时间点取皮肤检材,运用免疫组化和图像分析技术,检测血管内皮E-选择素的表达规律。结果在活体切创组中,伤后1hE-选择素在血管内皮细胞内即呈阳性表达,持续至伤后7d,且随时间变化呈规律性表达。在正常对照及死后切创组未见阳性表达。死后-20％稳定性组各时间点E-选择素表达与死后即刻比较无显著性差异（P〉0．05）。25％各时间点与死后即刻比较有显著性差异（P〈0．01）。结论E-选择素在创伤后血管内皮细胞内特异性的表达具有时序性规律,且低温条件下稳定性较好。     

The application of histochemical methods to the age evaluation of skin wounds: experimental study in rabbits.

Enzyme histochemical methods allow determination of wound age, especially in the range of a few hours, and are used to distinguish between postmortem and antemortem skin wounds. The methods are based on the determination of the presence and changes of the enzyme reaction in the wound area. Increased activity of nonspecific esterases was observed approximately 1 hour after wounding and was followed by an increase in adenosine triphosphatase at approximately 2 hours and alkaline phosphatase at approximately 3.5 hours. Maximum enzyme activity was reached for nonspecific esterases at 24 hours, for adenosine triphosphatase at 20 hours, and for alkaline phosphatase at 32 hours after wounding.