供腎零點(diǎn)活檢在腎移植應(yīng)用的研究
發(fā)布時(shí)間：2018-09-07 12:55
【摘要】：移植前供腎零點(diǎn)活檢能最直觀了解供腎組織結(jié)構(gòu)病變情況,能為臨床移植醫(yī)生對(duì)供腎質(zhì)量評(píng)估、受者的抉擇、供腎臨床分配及術(shù)后臨床個(gè)體化治療策略的提供主要的組織學(xué)依據(jù)。第一章公民逝世后器官捐獻(xiàn)供腎組織Remuzzi評(píng)分對(duì)移植效果的影響目的:通過(guò)分析公民死亡后器官捐獻(xiàn)供腎零點(diǎn)活檢組織的組織學(xué)病變,采用Remuzzi評(píng)分系統(tǒng),了解供腎組織不同病變程度對(duì)移植效果的影響。方法:1、研究對(duì)象于2015年1月至2015年12月在我院行公民死亡后器官捐獻(xiàn)腎移植術(shù)患者,并且術(shù)前供腎行零點(diǎn)活檢,符合以下標(biāo)準(zhǔn)則納入本研究:(1)、供腎組織病理檢查每張切片必須同時(shí)包含腎小球、腎小管、腎間質(zhì)及腎小血管四部分;(2)為了充分評(píng)估供腎組織病變,每張切片腎小球數(shù)量必須大于25個(gè);(3)受者至少隨訪6個(gè)月以上;(4)供受者均為成人,受者為成人單腎移植;2、研究方法獲取的供腎組織按照常規(guī)方法進(jìn)行固定、脫水、石蠟包埋、切片及Masson's染色,基于腎臟病理Masson's染色形態(tài)學(xué)特征,采用Remuzzi評(píng)分系統(tǒng)對(duì)供腎組織的腎小球硬化、腎小管萎縮、腎血管損傷及腎間質(zhì)纖維化病變程度進(jìn)行評(píng)分,各個(gè)部分得分相加得出整個(gè)腎臟的總體得分。3、實(shí)驗(yàn)分組根據(jù)供腎病理的總得分分為兩組,評(píng)分≤3為低分組,≥4為高分組.4、觀察指標(biāo)根據(jù)兩組供腎腎移植受者進(jìn)行術(shù)后隨訪1年,分析比較兩組供腎腎移植術(shù)后移植腎功能延遲恢復(fù)(delayed graft function,DGF)、DGF恢復(fù)時(shí)長(zhǎng)、術(shù)后3、6、12 個(gè)月腎小球率過(guò)濾(estimated glomerular filtration rate,eGFR)等相關(guān)指標(biāo)的差異。5、統(tǒng)計(jì)方法統(tǒng)計(jì)分析采用SPSS 20.0統(tǒng)計(jì)軟件。資料統(tǒng)計(jì)量采用均數(shù)±標(biāo)準(zhǔn)差(x±s)及百分比,組間均值差異性比較使用兩獨(dú)立樣本t檢驗(yàn),組間率的差異性比較使用χ2檢驗(yàn)或Fisher確切概率法,P0.05為差異有統(tǒng)計(jì)學(xué)意義。結(jié)果:供腎及相應(yīng)受者共62例納入本研究,供腎病變分值范圍為0-5分,低分組44例,高分組18例,總體DGF為30.6%,兩組間供腎對(duì)應(yīng)供者年齡、平均住院時(shí)長(zhǎng)及受者術(shù)后DGF發(fā)生率比較有顯著性差異(P0.05),術(shù)后DGF恢復(fù)時(shí)長(zhǎng)及1年內(nèi)腎小球率過(guò)濾兩組間比較均無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:供腎組織病變程度越重,受者腎移植術(shù)后DGF發(fā)生率越高,平均住院時(shí)間越長(zhǎng)。第二章非線性光學(xué)顯微成像技術(shù)在供腎質(zhì)量評(píng)估的應(yīng)用目的:探究二次諧波結(jié)合雙光子激發(fā)熒光的非線性光學(xué)顯微成像技術(shù)在供腎質(zhì)量評(píng)估。方法:研究對(duì)象:2015年1月至2016年5月在本移植中心行成人供死亡后器官捐獻(xiàn)供腎零點(diǎn)活檢組織。實(shí)驗(yàn)方法:零點(diǎn)活檢組織標(biāo)本經(jīng)固定、石蠟包埋后,每個(gè)標(biāo)本連續(xù)性切片兩張,每張5um,一張行傳統(tǒng)方法馬松三色染色,每例切片在高倍鏡下(200×)隨機(jī)觀察10個(gè)互不重疊腎小管間質(zhì)視野并拍照,另一張行二次諧波結(jié)合雙光子激發(fā)熒光顯微成像,采用Remuzzi評(píng)分系統(tǒng)分別對(duì)供腎組織四種組件進(jìn)行形態(tài)學(xué)半定量評(píng)分,即小管萎縮、間質(zhì)纖維化、腎小球硬化、血管損傷及供腎整體得分,用K系數(shù)描述SHG/TPEF方法對(duì)各個(gè)組件和總體評(píng)分與傳統(tǒng)Masson染色評(píng)估效果的一致性情況。統(tǒng)計(jì)方法:采用SPSS 20.0統(tǒng)計(jì)軟件進(jìn)行統(tǒng)計(jì)分析,用McNemar-Bowker檢驗(yàn)Masson染色和SHG/TPEF兩種方法算出的供腎組織各組件損傷評(píng)分構(gòu)成比的差異,以PO.05為有統(tǒng)計(jì)學(xué)差異。一致性檢驗(yàn)采用Kappa檢驗(yàn),當(dāng)kappa值0.75代表一致性好,0.40≤kappa值≤O.75代表一致性好,kappa值0.40代表一致性差。結(jié)果:非線性光學(xué)對(duì)供腎活檢組織有高分辨率的組織細(xì)胞成像,能對(duì)供腎組織質(zhì)量評(píng)估的腎小球、腎小管及血管有較好的形態(tài)學(xué)成像。具有不需要經(jīng)過(guò)染色、標(biāo)記,操作步驟簡(jiǎn)單、耗時(shí)短特點(diǎn),采用Remuzzi評(píng)分系統(tǒng)分別對(duì)供腎組織小管萎縮、間質(zhì)纖維化、腎小球硬化、血管損傷及供腎整體的半定量評(píng)分與Masson染色相比,差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05),在評(píng)估效果上與Masson染色評(píng)估效果一致性系數(shù):腎小球硬化k=0.895,間質(zhì)纖維化k=0.859,腎小管萎縮k=0.819,血管損傷k=0.802,總體情況k=0.914,全部一致性K系數(shù)大于0.75。結(jié)論:1、非線性光學(xué)顯微成像技術(shù)在供腎質(zhì)量評(píng)估效果與Masson染色評(píng)估效果相當(dāng);2、操作步驟簡(jiǎn)單、耗時(shí)短,具有運(yùn)用于移植前供腎質(zhì)量評(píng)估價(jià)值;3、對(duì)腎臟組織形態(tài)學(xué)上有較高分辨率。
[Abstract]:Zero-point biopsy of donor kidney before transplantation is the most intuitive way to understand the pathological changes of donor kidney. It can provide the main histological basis for clinicians to evaluate the quality of donor kidney, the choice of recipient, the clinical distribution of donor kidney and the clinical individualized treatment strategy after transplantation. Objectives: To analyze the histological changes of donor kidney after death and to evaluate the effect of different pathological changes of donor kidney by Remuzzi scoring system. Zero-point biopsy of the donor kidney was performed before operation, and the following criteria were included in this study: (1) Each section of the donor kidney must contain four parts: glomerulus, tubule, renal interstitium and renal vessels; (2) In order to fully evaluate the pathological changes of the donor kidney, the number of glomeruli in each section must be be greater than 25; (3) The recipient must be followed up for at least 6 months. (4) Donors and recipients were both adults and recipients were adult single kidney transplantation; (2) The donor kidney tissues obtained by the research method were fixed, dehydrated, paraffin embedded, sliced and Masson's stained according to the routine method. Based on the morphological characteristics of renal pathology Masson's staining, Remuzzi scoring system was used for glomerulosclerosis, renal tubular atrophy, renal vascular lesion in the donor kidney tissues. According to the total score of the donor kidney, the experimental group was divided into two groups. The score (< 3) was low and (> 4) was high. 4. The observation indexes were followed up for 1 year after operation according to the two groups. The difference of correlative indexes such as delayed graft function (DGF), DGF recovery time, estimated glomerular filtration rate (eGFR) at 3, 6 and 12 months after renal transplantation was analyzed by SPSS 20.0 statistical software. Results: A total of 62 donors and corresponding recipients were included in the study. The variational range of renal disease was 0-5, 44 in the low group, 18 in the high group and 30.6% in the total DGF. There were significant differences in donor age, average length of hospital stay and incidence of DGF between the two groups (P Chapter 2 Application of Nonlinear Optical Microscopic Imaging in Donor Quality Assessment Objective: To explore the application of second harmonic generation combined with two-photon excitation fluorescence in donor quality assessment. Methods: Zero-point biopsy specimens were fixed, paraffin-embedded, two consecutive sections of each specimen, each 5 um, one of which was stained with traditional Masson trichrome staining. Each specimen was randomly observed and photographed at high magnification (200 x) for 10 non-overlapping tubulointerstitial visual fields, and the other was subjected to second harmonic generation combined with two-photon excitation. Remuzzi scoring system was used to evaluate the morphology of four components of donor kidney, namely, tubular atrophy, interstitial fibrosis, glomerulosclerosis, vascular injury and overall donor kidney score. K coefficient was used to describe the consistency of SHG/TPEF scoring with traditional Masson staining. Conditions. Statistical methods: SPSS 20.0 statistical software was used for statistical analysis. McNemar-Bowker test was used to examine the difference of the component ratios of injury scores calculated by Masson staining and SHG/TPEF. PO.05 was used as the statistical difference. Results: Nonlinear optics had high resolution histocytography for renal biopsy tissues and good morphological imaging for glomeruli, tubules and blood vessels. It had the characteristics of no staining, marking, simple operation and short time-consuming. There was no significant difference between the Muzzi scoring system and Masson staining in terms of tubular atrophy, interstitial fibrosis, glomerulosclerosis, vascular injury, and the semi-quantitative score of the whole donor kidney (p0.05). The consistency coefficient between the evaluation results and Masson staining was: glomerulosclerosis k = 0.895, interstitial fibrosis k = 0.859, tubular fibrosis k = 0.859. Atrophy k = 0.819, vascular injury k = 0.802, overall k = 0.914, all consistency K coefficient greater than 0.75. Conclusion: 1, the effect of non-linear optical microscopy in the evaluation of donor kidney quality is equivalent to that of Masson staining evaluation; 2, simple procedures, time-consuming, with the value of pre-transplantation evaluation of donor kidney quality; 3, on the morphology of kidney tissue; It has higher resolution.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R699.2
本文編號(hào)：2228322
[Abstract]:Zero-point biopsy of donor kidney before transplantation is the most intuitive way to understand the pathological changes of donor kidney. It can provide the main histological basis for clinicians to evaluate the quality of donor kidney, the choice of recipient, the clinical distribution of donor kidney and the clinical individualized treatment strategy after transplantation. Objectives: To analyze the histological changes of donor kidney after death and to evaluate the effect of different pathological changes of donor kidney by Remuzzi scoring system. Zero-point biopsy of the donor kidney was performed before operation, and the following criteria were included in this study: (1) Each section of the donor kidney must contain four parts: glomerulus, tubule, renal interstitium and renal vessels; (2) In order to fully evaluate the pathological changes of the donor kidney, the number of glomeruli in each section must be be greater than 25; (3) The recipient must be followed up for at least 6 months. (4) Donors and recipients were both adults and recipients were adult single kidney transplantation; (2) The donor kidney tissues obtained by the research method were fixed, dehydrated, paraffin embedded, sliced and Masson's stained according to the routine method. Based on the morphological characteristics of renal pathology Masson's staining, Remuzzi scoring system was used for glomerulosclerosis, renal tubular atrophy, renal vascular lesion in the donor kidney tissues. According to the total score of the donor kidney, the experimental group was divided into two groups. The score (< 3) was low and (> 4) was high. 4. The observation indexes were followed up for 1 year after operation according to the two groups. The difference of correlative indexes such as delayed graft function (DGF), DGF recovery time, estimated glomerular filtration rate (eGFR) at 3, 6 and 12 months after renal transplantation was analyzed by SPSS 20.0 statistical software. Results: A total of 62 donors and corresponding recipients were included in the study. The variational range of renal disease was 0-5, 44 in the low group, 18 in the high group and 30.6% in the total DGF. There were significant differences in donor age, average length of hospital stay and incidence of DGF between the two groups (P Chapter 2 Application of Nonlinear Optical Microscopic Imaging in Donor Quality Assessment Objective: To explore the application of second harmonic generation combined with two-photon excitation fluorescence in donor quality assessment. Methods: Zero-point biopsy specimens were fixed, paraffin-embedded, two consecutive sections of each specimen, each 5 um, one of which was stained with traditional Masson trichrome staining. Each specimen was randomly observed and photographed at high magnification (200 x) for 10 non-overlapping tubulointerstitial visual fields, and the other was subjected to second harmonic generation combined with two-photon excitation. Remuzzi scoring system was used to evaluate the morphology of four components of donor kidney, namely, tubular atrophy, interstitial fibrosis, glomerulosclerosis, vascular injury and overall donor kidney score. K coefficient was used to describe the consistency of SHG/TPEF scoring with traditional Masson staining. Conditions. Statistical methods: SPSS 20.0 statistical software was used for statistical analysis. McNemar-Bowker test was used to examine the difference of the component ratios of injury scores calculated by Masson staining and SHG/TPEF. PO.05 was used as the statistical difference. Results: Nonlinear optics had high resolution histocytography for renal biopsy tissues and good morphological imaging for glomeruli, tubules and blood vessels. It had the characteristics of no staining, marking, simple operation and short time-consuming. There was no significant difference between the Muzzi scoring system and Masson staining in terms of tubular atrophy, interstitial fibrosis, glomerulosclerosis, vascular injury, and the semi-quantitative score of the whole donor kidney (p0.05). The consistency coefficient between the evaluation results and Masson staining was: glomerulosclerosis k = 0.895, interstitial fibrosis k = 0.859, tubular fibrosis k = 0.859. Atrophy k = 0.819, vascular injury k = 0.802, overall k = 0.914, all consistency K coefficient greater than 0.75. Conclusion: 1, the effect of non-linear optical microscopy in the evaluation of donor kidney quality is equivalent to that of Masson staining evaluation; 2, simple procedures, time-consuming, with the value of pre-transplantation evaluation of donor kidney quality; 3, on the morphology of kidney tissue; It has higher resolution.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R699.2
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