【摘要】： 1.大鼠腦組織缺血后Ngb mRNA表達(dá)與缺血時間推斷的研究 目的:探討大鼠腦組織缺血后不同時間腦紅蛋白(Neuroglobin,Ngb)mRNA表達(dá),旨為法醫(yī)病理學(xué)實踐中推斷損傷時間提供科學(xué)的參考依據(jù)。 方法采用夾閉雙側(cè)頸總動脈法建立Wistar大鼠腦組織缺血模型,提取大鼠在腦缺血0min(對照組)、1min、5min、10min、30min、60min時腦組織中的總RNA,經(jīng)質(zhì)量評價后,無降解的RNA進(jìn)行一步法逆轉(zhuǎn)錄-聚合酶鏈?zhǔn)椒磻?yīng)(one step RT-PCR),以GAPDH為內(nèi)參照,通過針對Ngb mRNA、GAPDH mRNA的特異性引物進(jìn)行擴增,對擴增后的產(chǎn)物進(jìn)行1.5%的瓊脂糖凝膠電泳,然后利用凝膠圖像分析技術(shù)檢測Ngb mRNA和內(nèi)參GAPDHmRNA的表達(dá),并將二者的比值(Ngb mRNA/GAPDH mRNA)與缺血間隔時間進(jìn)行相關(guān)性分析。 結(jié)果:(1)缺血后,Ngb mRNA在大鼠腦組織有明確而大量的表達(dá)。 (2)腦缺血后Ngb mRNA表達(dá)水平迅速升高,提示Ngb對缺氧反應(yīng)非常敏感,可能對神經(jīng)元起保護(hù)作用。 (3)Ngb mRNA的表達(dá)水平與缺血間隔時間有一定的相關(guān)性:缺血1min達(dá)高峰,與對照組比較差異有顯著性(P＜0.01);缺血5min表達(dá)迅速降低,至10min仍維持在較低水平,與對照組相比差異無顯著性(P＞0.05):隨缺血時間延長,表達(dá)水平再度逐漸增強,與對照組相比差異有顯著性(P＜0.01)。 結(jié)論:大鼠腦缺血缺氧后腦紅蛋白的轉(zhuǎn)錄水平迅速增高,說明腦紅蛋白對缺氧的反應(yīng)非常敏感,可能參與了腦對缺氧的保護(hù)性反應(yīng)。大鼠腦組織缺血后Ngb mRNA的表達(dá)隨缺血時間的變化而有所不同,其與腦缺血間隔時間存在明顯相關(guān)性,可在法醫(yī)實踐中用于早期腦缺血時間的推斷。 2.人體腓骨性別判別分析 目的:探討應(yīng)用腓骨進(jìn)行性別判別的方法,建立腓骨性別判別的函數(shù)判別式。 方法:采用腓骨5項測量指標(biāo),用人體測量儀器,按體質(zhì)人類學(xué)測量方法,對國內(nèi)9個不同省區(qū)、已知確切性別的305具尸骨(男274例,女31例,年齡范圍18-84歲)的左、右側(cè)共610根腓骨進(jìn)行測量。將所測量數(shù)據(jù)作為變量輸入計算機;使用社會科學(xué)統(tǒng)計軟件包11.5(SPSS11.5),對左、右側(cè)腓骨和男、女腓骨的統(tǒng)計學(xué)數(shù)據(jù)分別依據(jù)側(cè)別和性別進(jìn)行分組,然后進(jìn)行兩獨立樣本的t檢驗;篩選出具有顯著性意義的測量指標(biāo),用Fisher判別分析法,建立腓骨性別判定的函數(shù)判別式。 結(jié)果:(1)男性或女性腓骨的左、右側(cè)指標(biāo)測量值的側(cè)別差異均無統(tǒng)計學(xué)意義(P＞0.05),可以選擇單側(cè)進(jìn)行研究。 (2)腓骨5項指標(biāo)測量值的性別差異均具有極顯著意義(P＜0.01),可以利用其進(jìn)行性別判別。 (3)獲得腓骨5個單指標(biāo)函數(shù)判別式及1個多指標(biāo)函數(shù)判別式,前者綜合判別率在61.64%～75.74%,后者綜合判別率達(dá)82.62%。 結(jié)論:腓骨單指標(biāo)和多指標(biāo)函數(shù)判別式可用于國人的性別判定,多指標(biāo)函數(shù)判別式更具應(yīng)用價值。
[Abstract]:1. Study on the expression of Ngb mRNA and the inference of ischemic time after cerebral ischemia in rats objective: to investigate the expression of brain red protein (Neuroglobin,Ngb) mRNA at different time after cerebral ischemia in rats. The purpose of this study is to provide a scientific reference for inferring the time of injury in the practice of forensic pathology. Methods the cerebral ischemia model of Wistar rats was established by clamping bilateral common carotid artery method. The total RNA, in brain tissue of rats with cerebral ischemia 0min (control group) was extracted at 10 min, 30 min and 60 min after 1 min, 5 min, 30 min and 60 min, and the total RNA, in brain tissue was evaluated. One-step reverse transcription-polymerase chain reaction (one step RT-PCR) of non-degraded RNA was carried out, and GAPDH was used as internal reference to amplify Ngb mRNA,GAPDH mRNA by specific primers. The amplified products were subjected to 1.5% agarose gel electrophoresis, and then the expression of Ngb mRNA and internal reference GAPDHmRNA was detected by gel image analysis, and the correlation between the ratio of Ngb mRNA/GAPDH mRNA and ischemic interval was analyzed. Results: (1), Ngb mRNA was clearly and heavily expressed in rat brain tissue after ischemia. (2) the expression of Ngb mRNA increased rapidly after cerebral ischemia, suggesting that Ngb is very sensitive to hypoxia and may play a protective role in neurons. (3) there was a certain correlation between the expression level of) Ngb mRNA and the interval between ischemia and ischemia: the peak of ischemic 1min reached the peak, which was significantly different from that of the control group (P < 0.01). The expression of ischemic 5min decreased rapidly and remained at a low level until 10min, which was not significantly different from that of the control group (P > 0.05). With the prolongation of ischemia time, the expression level increased gradually again. There was significant difference between the two groups (P < 0.01). Conclusion: the transcription level of brain red protein increases rapidly after cerebral ischemia and hypoxia in rats, which indicates that brain red protein is very sensitive to hypoxia and may be involved in the protective response of brain to hypoxia. The expression of Ngb mRNA after cerebral ischemia in rats is different with the change of ischemic time, and it is significantly correlated with the interval of cerebral ischemia, which can be used to infer the time of early cerebral ischemia in forensic practice. 2. Objective: to explore the method of sex discrimination of fibula and to establish a functional discriminant of sex discrimination of fibula. Methods: the left of 305 corpses of known sex (274males and 31 females, age range 18 鈮，

本文編號：2480579