本文選題：死亡時(shí)間推斷 + 肝糖原�。� 參考：《四川大學(xué)》2007年碩士論文

【摘要】： 研究背景及目的 死亡時(shí)間的推斷的研究開始于1800年(Nysten)，但是由于人體死亡之后受多種因素的影響，因此，一直無(wú)法用單一指標(biāo)來推測(cè)死亡時(shí)間，幾個(gè)世紀(jì)以來，中外學(xué)者進(jìn)行了不斷探索。但時(shí)至今日，仍然不能準(zhǔn)確推斷。然而，對(duì)于解決實(shí)際工作的問題最直接有效的辦法，是采用多種方法進(jìn)行綜合分析推斷，利用超生反應(yīng)進(jìn)行死亡時(shí)間推斷，多數(shù)是用形態(tài)學(xué)觀察和機(jī)械性電興奮性方面，而肝臟組織內(nèi)能量物質(zhì)變化與死亡時(shí)間推斷方面的研究，能夠直接用于法醫(yī)學(xué)方面的成果較少，已有的文獻(xiàn)多見于運(yùn)動(dòng)醫(yī)學(xué)和肝臟活體移植方面。 因此，我們?cè)谇叭搜芯康幕A(chǔ)上，本實(shí)驗(yàn)通過建立不同動(dòng)物死亡模型，采用宏觀觀察和微觀分析相結(jié)合的方法，從死亡后肝臟組織內(nèi)糖原，磷酸果糖激酶及腺苷酸磷酸變化的角度探討推斷死亡時(shí)間的有效方法，期待能為法醫(yī)學(xué)實(shí)踐提供有用的參考。 材料與方法 健康Sprague-Dawley大鼠180只，體重250±20克，分為頸椎拉斷斷處死組與窒息死亡失血性休克死亡組，每組60只，分別于死亡0min，30min，1h，1.5h，2h，3h，4h，6h，8h，12h，18h，24h時(shí)間點(diǎn)取肝臟在—20℃冰箱內(nèi)冷凍，所有肝臟標(biāo)本均分為二份，一份作冰凍切片進(jìn)行PAS(含對(duì)照)及磷酸果糖激酶免疫組化染色；另一份作ATP，ADP，AMP含量分析。 每個(gè)時(shí)間點(diǎn)五只動(dòng)物取其平均值作檢驗(yàn)結(jié)果。從以下幾個(gè)方面進(jìn)行檢測(cè)。 (1)肝臟糖原的變化，肝臟組織作PAS染色并做計(jì)算機(jī)圖象分析，利用灰度值的變化規(guī)律推測(cè)死亡后肝臟內(nèi)的糖原的改變情況，從而推斷死亡時(shí)間及死亡原因。 (2)免疫組化染色檢測(cè)磷酸果糖激酶—2，并將染色結(jié)果用計(jì)算機(jī)圖象分析技術(shù)，觀察肝臟內(nèi)表達(dá)情況和變化規(guī)律。 (3)利用高效液相色譜技術(shù)對(duì)肝臟內(nèi)ATP，，ADP，AMP的含量進(jìn)行測(cè)定。 結(jié)果 1．大鼠肝臟內(nèi)糖原含量隨死后時(shí)間的不同而呈線性下降。提示可以作為死亡時(shí)間推斷的一個(gè)參考指標(biāo)。 2．大鼠肝臟內(nèi)磷酸果糖激酶-2的免疫組化染色顯示，其灰度值改變有一定規(guī)律性，可作死亡時(shí)間推斷的參考指標(biāo)。 3．大鼠肝臟內(nèi)ATP，ADP，AMP的含量呈規(guī)律性變化，而且不同死亡原因，其肝臟組織內(nèi)的腺苷酸磷酸含量有差異，提示可以用來推斷死亡時(shí)間，其中以ATP含量變化最具有應(yīng)用價(jià)值。
[Abstract]:Research background and purposeThe study on the estimation of the time of death began in 1800, but because of the influence of many factors after the death of the human body, it has been impossible to estimate the time of death by a single index. For several centuries, Chinese and foreign scholars have been exploring continuously.But today, still can not accurately infer.However, the most direct and effective way to solve the practical problems is to use a variety of methods for comprehensive analysis and inference, and to infer the time of death by using supergenetic reactions, most of which are based on morphological observation and mechanical electrical excitability.However, there are few studies on the changes of energy and time of death in liver tissue, which can be directly used in forensic medicine, and the existing literatures are mostly found in sports medicine and liver transplantation.Therefore, on the basis of previous studies, we established different animal death models and adopted the method of macroscopical observation and microanalysis to analyze glycogen in liver tissue after death.From the point of view of the changes of phosphofructokinase and adenylate phosphoric acid, an effective method for estimating the time of death is discussed, which is expected to provide a useful reference for forensic practice.Materials and methods180 healthy Sprague-Dawley rats, weighing 250 鹵20 g, were divided into two groups, 60 in each group. Each group was divided into two groups, 60 rats in each group. The liver was frozen in refrigerator at -20 鈩

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