本文關(guān)鍵詞： 法醫(yī)學(xué) 年齡推斷 southern雜交 自溶　出處：《山西醫(yī)科大學(xué)》2007年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的探討人外周血白細(xì)胞端粒DNA長(zhǎng)度與年齡、性別的關(guān)系,及組織自溶對(duì)southern雜交法測(cè)量結(jié)果的影響,以評(píng)價(jià)通過(guò)人外周血白細(xì)胞端粒長(zhǎng)度推斷個(gè)體年齡的法醫(yī)學(xué)應(yīng)用價(jià)值。 方法1.端粒—年齡相關(guān)性研究:收集97例健康人外周血,按年齡分為20-40歲組、41-60歲組和＞61歲組。酚-氯仿法抽提人外周血白細(xì)胞基因組DNA,southern雜交方法測(cè)量端粒DNA限制性酶切片斷(TRF)長(zhǎng)度。2.組織自溶對(duì)平均端粒長(zhǎng)度測(cè)得值的影響:另取6例同年齡(27歲)同性別(男性)人外周血各3ml,將同一血樣本平均分成3份(每份1ml)分別于收集后、人工氣候箱(25℃、50%)內(nèi)放置24h、放置72h提取人外周血白細(xì)胞DNA,上述方法測(cè)出經(jīng)不同時(shí)間自溶后人外周血白細(xì)胞端粒DNA限制性酶切片斷長(zhǎng)度(TRF值)。 結(jié)果1.端粒—年齡相關(guān)性研究:不同年齡組測(cè)得的TRF值隨年齡增大依次遞減,通過(guò)SNK法檢驗(yàn)不同年齡組間TRF值差異,結(jié)果顯示差異顯著(P＜0.05)。對(duì)各年齡組中不同性別的TRF值進(jìn)行t檢驗(yàn),發(fā)現(xiàn)除20-40歲組人外周血白細(xì)胞TRF值的性別差異不顯著外(P＞0.05),其余各組TRF值的性別差異均有統(tǒng)計(jì)學(xué)意義(P＜0.05)。年齡與TRF值的相關(guān)線性關(guān)系良好(y=-0.054x+11.16 R2=0.64)。分別對(duì)不同性別組年齡與TRF值進(jìn)行線性分析發(fā)現(xiàn),男性為y=-0.069x+11.67(R2=0.71),女性為y=-0.39x+10.62(R2=0.69)。2.組織自溶對(duì)平均端粒長(zhǎng)度測(cè)得值的影響:SNK法對(duì)各自溶組間測(cè)得的TRF值進(jìn)行比較,發(fā)現(xiàn)24h組與收集后組測(cè)得端粒長(zhǎng)度差異不大(P＞0.05),72h組與即刻組和24h組都有顯著性差異(P＜0.05)。 結(jié)論1.端粒—年齡相關(guān)性研究:端粒限制性酶切片斷長(zhǎng)度隨個(gè)體年齡的增長(zhǎng)而縮短,端粒長(zhǎng)度的性別差異除20-40組外都有明顯的統(tǒng)計(jì)學(xué)意義,且不同性別端粒的縮短速度不同,男性縮短速度快于女性。2.組織自溶對(duì)平均端粒長(zhǎng)度測(cè)得值的影響:southern雜交法測(cè)得的端粒DNA限制性酶切片斷長(zhǎng)度結(jié)果受組織自溶影響變化較大。研究提示人外周血白細(xì)胞端粒DNA限制性酶切片斷可以作為法醫(yī)學(xué)年齡推斷的參考,但應(yīng)考慮組織自溶的影響。
[Abstract]:Objective to investigate the relationship between telomere DNA length and age, sex of human peripheral blood leukocytes, and the effect of autolysis on the results of southern hybridization, and to evaluate the value of forensic medicine in estimating individual age by using telomere length of human peripheral blood leukocytes. Methods 1.The study of telomere age correlation: the peripheral blood was collected from 97 healthy people. The length of telomere DNA restriction fragment length was measured by phenol-chloroform hybridization method. The mean telomere length was measured by tissue autolysis and the mean telomere length was measured by autolytic assay for the mean telomere length of human peripheral blood leukocytes by phenol-chloroform hybridization method, which was divided into two groups according to their age: 41-60 and > 61.The length of telomere restriction fragment was measured by phenol-chloroform hybridization. In addition, 3 ml of peripheral blood were taken from 6 patients of the same age and 27 years of age) of the same sex (male), and the same blood samples were divided into 3 parts (1 ml each) respectively after collection. The human peripheral blood leukocyte DNA was extracted at 25 鈩，

本文編號(hào)：1509046