纤维联接蛋白的析出与大鼠皮肤早期损伤的时间关系

采用免疫组化(PAP)方法,检测大鼠皮肤损伤区的FN,发现在生前创伤后15min,创壁FN即呈明确阳性;随着伤后经历时间的延长,创壁FN逐渐增多,并滑创壁呈条带状沉积;而在死后5min的创伤,创壁FN则呈阴性。本文为区别生前与死后皮肤创伤提出了一种新的方法。     

对330例体表创口与愈后瘢痕长度的对比观察

探讨体表创口与愈后瘢痕相互关系。在 330例不同年龄的伤者的不同部位的钝、锐器伤口 ,用精确的测量方法 ,在伤后 3d内、伤后 1个月、伤后 3个月进行测量 ,并计算创口愈合成为瘢痕的收缩率。在伤后 3个月面部瘢痕的收缩率为 15 4% ,躯干、四肢瘢痕收缩率为 13 6 % ,头皮瘢痕收缩率为 10 1%。不同致伤物形成的创口 ,在伤后 3个月时 ,不规则钝器创的瘢痕收缩率为 16 1% ,锐器创的瘢痕收缩率为 11 2 %。结果表明 ,创口在伤后 3个月时 ,面部瘢痕收缩率最大 ,头皮瘢痕收缩率最小。不规则钝器创的瘢痕收缩率值比锐器创的瘢痕收缩率值大。     

Connexin 43, angiotensin II, endothelin 1, and type III collagen alterations in heart of rats having undergone fatal electrocution

Death due to accidental electrocution occurs frequently. The aim of this study was to investigate alterations in cardiac connexin 43 (Cx43), angiotensin II (Ang II), endothelin 1 (ET-1), and type III collagen associated with fatal electrocution.Twenty-four Sprague-Dawley rats were divided into control, fatal electrocution (220 V, 50 Hz, 60 seconds), and electrical injury (220 V, 50 Hz, 60 seconds) groups. Animals were deeply anesthetized with sodium pentobarbital before each treatment, with the anode connected to the left foreleg and the cathode to the right hindleg, followed by cervical dislocation. Control animals received cervical dislocation alone. Immunohistochemical analysis was performed to evaluate the cardiac protein expression of Cx43, Ang II, ET-1, and type III collagen. Sections were analyzed by digital image analysis.The expression of Cx43 was significantly reduced after fatal electrocution, with the integrated optical density also lower when compared with control (P < 0.05). Expression of both Ang II and ET-1 was significantly increased after fatal electrocution, supported by integrated optical density when compared with control (P < 0.05). But no significant difference was found in type III collagen expression between the fatal electrocution group and the control group.In summary, cardiac protein expression of Cx43, Ang II, and ET-1 was found to be significantly altered with fatal electrocution, suggesting that these 3 proteins may be important underlying mechanisms of death during fatal electrocution. The current findings indicate that such alterations would be reflected in abnormal cardiac function and a possible cause of sudden death.     

New observations on cutaneous firearm wounds

The authors describe the aspect of the dermal surface in firearm wounds. In entrance wounds, the abraded area is rather evident, with loss of the papillary pattern and lacerations of the basement membrane. Exit wounds appear as irregularly lacerated interruptions, with protruding collagen fibers; the surrounding dermis has nearly undamaged papillae. Also described, still on the dermal surface, are the flame effect and the powder tattooing. Evident lesions of the basement membrane, produced by the grains of powder, can be observed. Particles [gunshot residues (GSR)] are often recognizable lying on the dermal surface or entrapped among the collagen fibers of the superficial dermis.     

Quantitative analysis of proinflammatory cytokines (IL-1beta, IL-6, TNF-alpha) in human skin wounds

Proinflammatory cytokines play an important role in the mediation of inflammation and trauma. They could be useful for the determination of vitality and wound age. In the present study, 144 human skin wounds due to sharp force were investigated. The material was collected during operations (N=96) and postmortem examinations (N=48). The wound age varied from several seconds or minutes to 9 days. Control skin was available in each individual. The tissue specimens were homogenized and extracted in a solution of PBS and protease inhibitors. Interleukin-1beta (IL-1beta), interleukin-6 (IL-6) and tumour necrosis factor alpha (TNF-alpha) were measured by quantitative ELISA analysis. Statistical evaluation was performed by the t-test using the quotients of levels (wound sample/control skin). In surgical specimens the cytokine levels revealed a clear tendency to increase with wound age. IL-1beta in early skin wounds (24 h, P<0.05). The quantitative analysis of proinflammatory cytokines in wound extracts can contribute to the determination of vitality and wound age, in particular in the very early post-traumatic interval (classic stab wounds).     

小鼠皮肤创伤愈合过程中IL-10在不同表达部位的组化定量研究

目的探讨小鼠皮肤创伤愈合过程中IL-10在不同表达部位的变化与损伤时间的关系。方法应用免疫组织化学技术和形态计量法,对小鼠不同时程皮肤切创组织中IL-10不同表达部位(表皮细胞及表皮下组织的浸润细胞)的变化进行研究。结果皮肤切创前后表皮层均有阳性表达,切创后1～3h表皮细胞阳性表达水平迅速升高,24h时降至较低水平,伤后48h表达水平再次升高,96h后逐渐回复到正常水平;表皮以下阳性部位主要是浸润的单核/巨噬细胞,损伤6h后阳性细胞在真皮、皮下组织及创口肉芽组织内逐渐增多,于伤后72h达最大值(51.41±3.12)%,96～168h阳性细胞数逐渐减少(29.38±2.64)%～(5.56±4.74)%。结论皮肤创伤修复过程中IL-10在不同表达部位的变化特点与皮肤损伤时间相关,检测其表达变化可望用于损伤时间推断。     

Expression of adhesion molecules in skin wounds: diagnostic value in legal medicine

The adhesion molecules identified in recent years can help improve the diagnosis of the wound age, especially of injuries with a short survival time. This is also indicative of the vitality of the wounds. The material investigated in the study originated from 465 skin wounds. The samples were taken from human autopsy material, during the surgical treatment of wounds (excision) of patients and from experimental incised wounds of mice. To judge the age of skin wounds the endothelial adhesion molecules were detected in paraffin sections after autoclaving and using the ABC technique. Human skin wounds: strong positive staining was observed of ICAM-1 1.5 h at the earliest and 3.5 days at the latest, for the P-selectin 3 min at the earliest and 7 h at the latest, for the E-selectin 1 h at the earliest and 17 days at the latest and for VCAM-1 3 h at the earliest and 3.5 days at the latest after the time of injury. The L-selectin was expressed constitutively. Mice skin wounds: strong positive immunohistochemical reactions were found as a rule earlier than in human skin wounds. The detection of an increased expression of ICAM-1, VCAM-1 and P- and E-selectins can improve the wound age assessment in injuries with short survival times.     

Peculiarities of the stab and cut wounds caused by blades of combat cold weapons

Design philosophy of blades of Russia-made combat cold arms and of inflicted-by-them stab-and-cut wounds were characterized on the basis of a comparative-and-evaluation study. Stereomicroscopy, histology examinations as well as X-ray-spectral and fluorescence analyses were made use of while investigating the wounds with subsequent processing of the results by discriminative analysis. Form, skewness and a graze type of the wound tubber as well as presence of "edge cuts" and of central graze wound were found to be the main distinctive wound signs. The study results ensure a full-value differential diagnosis of a type of the used cold-arm sample.     

Ruptured cerebral artery aneurysm and bacterial meningitis in a man with osteogenesis imperfecta

This report describes a 38-year-old man with osteogenesis imperfecta who died of a ruptured cerebral artery aneurysm and bacterial meningitis. He had multiple long bone fractures in the past, and approximately 4 months before death, he had surgery to relieve symptoms of basilar impression. The surgery was complicated by a postoperative wound infection. For the next 4 months, he had intermittent headaches and vomiting. He was found dead in his bed at home. At autopsy, he had a ruptured anterior communicating artery aneurysm and bacterial meningitis. Cerebrospinal fluid and blood cultures had growth of Staphylococcus aureus. Osteogenesis imperfecta is a disorder of type I collagen. Type I collagen is present in many tissues, including blood vessels. The etiology of cerebral artery aneurysm formation is multifactorial. Some patients with cerebral artery aneurysms have been shown to have abnormalities in type III collagen. There has not been a reported relationship made between abnormalities in type I collagen and aneurysms. Meningitis can also result in cerebral artery aneurysms, but they are usually due to Aspergillus or Mycobacterium species. The case we report is unique; cerebral artery aneurysm formation may have been due to osteogenesis imperfecta and/or bacterial meningitis.     

Dermal surface morphology in wound healing. An experimental scanning electron microscope study

The authors observed, with a scanning electron microscope, the dermal surfaces of experimental wounds, following the enzymatic digestion with trypsin of the dermo-epidermal junction. Cut lesions showed an early reconstitution of the basement membrane and progressive convergence of the margins. A peculiar arrangement of the dermal crests was also observed. In wounds with loss of substance, the complete coating by newly formed basement membrane took longer. There was an early deformation of the holes of hair follicles in the undamaged dermis surrounding the lesion, as well as the appearance of creases that moved toward the center. This process was over within a few days; then the lesion became very small and the holes of the hair follicles reappeared. The results suggest a force of traction bound toward the center of the lesion and acting on the surface, the effects of which on the basement membrane are most evident between the eighth and the 15th days.     

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。     

Simultaneous observation of catecholamine, serotonin and their metabolites in incised skin wounds of guinea pig.

In order to examine the vital reaction in wounds, catecholamines, serotonin (5-HT) and their metabolites in the incised skin wounds of guinea pigs were analyzed simultaneously by high-performance liquid chromatography (HPLC) using electrochemical detection (ECD). The principal changes in the levels of these compounds in vital wounds were as follows: a considerable decrease in norepinephrine (NE) content was observed 12-24 h after injury which persisted to at least 7 days. 3,4-Dihydroxyphenylacetic acid (DOPAC) decreased slightly for up to 30 min and then showed no significant difference compared with postmortem levels. Epinephrine and dopamine were barely detected by the HPLC-ECD method employed. 5-HT concentrations which showed an increase up to 24 h showed maximum levels 800 microns from the wound edge at 10 and 30 min after injury. 5-Hydroxyindoleacetic acid (5-HIAA) was significantly higher than the postmortem level over almost the entire period of these experiments. A 5-HIAA content of at least twice the postmortem level was observed 800 microns from the wound edge of a 10-min-old vital wound. Therefore, 5-HIAA is a likely candidate as a new marker for evaluating the vital reaction in wounds. The vital characteristics of NE, DOPAC, 5-HIAA and 5-HT in 10-min-old wounds persisted for up to 12 h at room temperature after death. These results suggest that the HPLC-ECD method used here is very useful for simultaneous examination of the vital reaction in wounds from the earliest to the later stages of the wound-healing period.     

A preliminary study on the changes of expression of PDGF-beta,PDGFR-beta,TGF-beta 1, TGFR,bFGF and its relationship with the wound age in wound healing

OBJECTIVE: To explore the relationship between the expression change of cytokines and the wound age during the healing process of rats skin wound. METHODS: Immunohistochemical and image-analysis methods were performed on vital skin wounds(after incision 0.5-168 h am) and postmortem damage(after incision 0.5-6 h pm). RESULTS: The expression of the cytokines PDGF-beta, PDGFR-beta, TGF-beta 1, and bFGF in the epithelial cells was already enhanced since 0.5 h am after damage and their strongest expression reaction was seen at 24-96 h am. In addition, the expression of PDGF-beta, PDGFR-beta, TGF-beta 1 and bFGF was also found in the macrophages and the fibroblasts of the granulation tissue, and the expression changes in the postmortem damage group showed that the skin tissue within 0.5-3 h after incision showed immunohistochemical changes but weakly expression and 3 h thereafter no any change was found. CONCLUSION: The expression characteristics of the above mentioned cytokines in wound repair should be related to the wound age and it reminds therefore that they may be used as immunohistochemical criteria for accurate determining the wound age.     

大鼠皮肤切创愈合过程中泛素的表达及其法医学意义

目的观察大鼠皮肤切创后泛素(ubiquitin)表达的变化规律。方法应用免疫组化方法,观察大鼠皮肤切创后1、3、6、12h和1、3、6、10、14d ubiquitin的表达情况,并用图像分析系统进行图像分析。结果正常对照组大鼠皮肤有低水平ubiquitin表达,切创后ubiquitin表达增加,伤后6d达峰值,伤后10d开始下降,伤后14d恢复到正常水平。结论ubiquitin可作为法医学损伤时间推断的有效指标。     

损伤经过时间的实验研究

作者在全身麻醉及无菌条件下,通过在大白鼠背部皮肤造成的切创创口,于伤后不同时间杀死.每组中1只于处死前8小时注射秋水仙碱以观察核分裂.取损伤处皮肤,经福尔马林固定、石蜡切片、HE染色、Gorden和Sweet染色(网状纤维)、Weigert染色(弹力纤维)、PTAH染色(纤维蛋白)和Van Gieson染色,于显微镜下观察.结果发现创口逐日缩小,但其中24小时创口较12小时创口有所扩大,伤后第6～7日创口呈突击式缩小.通过组织学变化观察,作者对创口收缩现象在判断损伤时间的意义及形成机理作了讨论.     

An Unusual Suicide Inside a Grave with Two Gunshot Wounds to the Head—The Psychological Approach

Abstract: In this report, the authors present a case of suicide committed by a 66-year-old man, inside a grave at the local cemetery, with two gunshot wounds to the head. A multiple-paged, handwritten suicide note was found in an army type bag inside the victim’s car, while a second one was found in his house. Medicolegal examination of the body revealed two typical contact gunshot entrance wounds and one exit wound to the head. Toxicological analysis of the femoral blood was negative for alcohol and drugs. The history, scene and autopsy findings, along with further police investigation, indicated an obvious case of suicide. The literature is reviewed and discussed. We report this case as the place of suicide is extremely uncommon and as there is no mention of a gunshot suicide inside a grave in the current literature.     

Unusual shotgun injury. Gas blowout of anterior head region

During a 6-month period, we encountered two examples of tight contact wounds of the temple region which were associated with ovoid, fairly regular, perforating wounds of the skull and overlying skin. These wounds were located between the eyes with the upper border near the glabella and the lower border near the level of the lower eyelids. There was no evidence that these wounds were entry wounds and in both cases the projectiles were still within the cranial cavity. Therefore, they were interpreted as gas blowout wounds. No similar cases have been found in the literature. Law enforcement personnel and pathologists should be aware of this rather rare blowout type of shotgun injury since it may be subject to various misinterpretations, including that of an entry wound.     

兔不同时间切创中肌细胞核DNA的定量测定

为了使损伤时间推断手段更加精确化,为损伤时间推断提供新的方法,我们利用显微分光光度计测定兔皮肤切创创口内肌细胞核DNA 含量,结果发现:正常肌细胞核DNA 含量为18. 420Au,而损伤后创外周区内肌细胞核的DNA 含量立即迅速下降。伤后1h 为17. 274Au;伤后3h 为8. 672Au,达最低水平,随着损伤及经过时间的延长,DNA 含量又逐渐回升,伤后6、8、12,24h 的含量分别为12. 563、13. 608、14. 743,16. 448Au。上述结果说明,损伤区域内肌细胞核内DNA 的含量与损伤时间有明显关系,DNA定量分析可作为损伤时间推断的一个重要根据。     

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.     

在大鼠皮肤切创愈合过程中TGF-β_1基因及蛋白表达变化与损伤时间关系的实验研究

了解细胞因子在创伤及修复过程中的表达变化与损伤时间的关系。初步调查生前不同造创时间 ( 0 5h～16 8h)皮肤创缘组织在修复过程中TGF β1mRNA和蛋白质表达变化 ,并与死后伤 ( 0 5h～ 6h)上述变化进行比较。结果表明 ,在生前伤 ,TGF β1在上皮细胞的表达于损伤后 0 5h开始增强 ,以 2 4h～ 96h反应最强 ,除上皮细胞内表达外 ,还在肉芽组织的巨噬细胞和成纤维细胞表达。TGF β1的免疫印迹 (WesternBlot)分析表明 ,TGF β1蛋白表达的峰值在 16 8h ;TGF β1斑点杂交和原位杂交结果显示 ,mRNA的表达在伤后 3h开始 ,6h后明显 ,96h达峰值。死后损伤组在 0 5～ 3h内有免疫组化的弱～中度表达 ,但无mRNA的表达 ,3h后则均无任何表达。TGF β1在修复过程中的一些规律性变化及特点反映了与损伤时间的关系 ,可作为精确推断损伤时间的分子生物学标志。