Cell-Biological Study on Medicolegal Diagnosis of Injury

在本研究中，我们从细胞死亡的两种机制，即坏死和凋亡，对损伤引起的细胞死亡进行了细胞生物学研究。为了对RTS的肝脏进行加热，我们新开发了一种内置PID电子调节系统的加热装置。麻醉下剖腹取肝，分别在37ﾟC、50ﾟC和65ﾟC下加热5min。用一根加热的针连接到装置上，然后在给定的时间内存活下来。通过苏木精-伊红(HE)染色、原位末端标记法(TUNEL)染色和DNA琼脂糖凝胶电泳法，分析了热损伤组织在存活过程中细胞死亡的表现，结果表明，在所有温度下，细胞都出现了凋亡，并在热后2小时和10小时左右出现双峰模式。在37ﾟ℃的受热温度下，整个受热区域都出现了点状的第一个凋亡峰。当Li…在50ﾟC和65ﾟC温度下加热的VER较多，先出现在受热区域的外缘，然后向区域中心进发，HE染色显示，在损伤组织与正常组织的交界处，染色质凝集明显。这些结果与37ﾟC的结果不同。第一个峰的细胞凋亡似乎是由温度的直接作用引起的。第二个高峰期，在所有温度下，细胞凋亡从损伤区域的外缘向中心进展，其进展的时间进程与中性粒细胞向损伤区域的渗透几乎是同步的。从显示细胞凋亡的组织中提取的DNA用于琼脂糖凝胶电泳时，观察到了梯状结构。损伤组织中的坏死在时间和部位上与细胞凋亡的出现形式一致，坏死和细胞凋亡的程度几乎相同，直至出现第二个细胞凋亡高峰。加热24小时后，凋亡细胞被吞噬，仅留下坏死组织。肿瘤坏死和细胞凋亡的出现似乎是由于中性粒细胞来源的肿瘤坏死因子α的作用，因为在电子显微镜下证实了肿瘤坏死因子α的产生。在本研究中，细胞生物学证据表明，细胞凋亡与损伤组织中的细胞死亡密切相关。较少

In this study, we investigated cell-biologically the cell death generating by injuries, on the basis of two mechanisms of the cell death, these are necrosis and apoptosis.To heat the liver of rts, a heat device having a built-in PID electron regulater system was newly developed by us. The liver of rats laparotomized under anesthesia was heated at 37ﾟC,50ﾟC and 65ﾟC for 5 min. with a heated needle connected to the device, and thereafter the rats were survived with in the given time. Appearance of the cell death in the tissue injured by heat was analyzed, in progress of the survival time, by means of staining of the tissue with hematoxylin-eosin (HE) and TUNEL method and agarosegel electrophoresis of DNA extracted from the tissue injured.Consequently, apoptosis appeared, in all heat temperatures, with bimodal pattern showing peak at about 2 and 10 hours after heat. Apoptosis in the first peak dottedly appeared, in heat temperature at 37ﾟC,in the whole region affected by heat. When the li … More ver was heated at 50ﾟC and 65ﾟC,it first appeared on outer edge in the region affected by heat, and then made progress to the center of the region, condensation of chromatin was, in HE staining, remarkable at border between the injured and normal tissues. These findings were different from those in 37ﾟC.It seems that apoptosis inthe first peak is caused by direct action of temperature. Apoptosis in the second peak, in all heat temperature, made progress from outer edge of the injured region to the center, time course of progression of the apoptosis was almost concurrent with that of infiltration of neutrophil to the injured region. Ladder formation was observed, when DNA extracted from the tissue showing apoptosis was applied for agarosegel electrophoresis. Necrosis in the injured tissue appeared in the same pattern with appearance of apoptosis in time course and on location, necrosis and apoptosis were observed in almost same degree up to appearance of the second peak of apoptosis. After 24 hours of heat, apoptosis cells were phagocytosed, only necrotic tissue remained. It seems that appearance of necrosis and apoptosis is caused by action of TNF alpha derived from neutrophils, since generation of TNF alpha was confirmed electron microscopically.In this study, it became cell-biologically evident that apoptosis is intensely concerned with cell death in the injury tissue. Less