供肾零点活检在肾移植应用的研究
发布时间：2018-09-07 12:55
【摘要】：移植前供肾零点活检能最直观了解供肾组织结构病变情况,能为临床移植医生对供肾质量评估、受者的抉择、供肾临床分配及术后临床个体化治疗策略的提供主要的组织学依据。第一章公民逝世后器官捐献供肾组织Remuzzi评分对移植效果的影响目的:通过分析公民死亡后器官捐献供肾零点活检组织的组织学病变,采用Remuzzi评分系统,了解供肾组织不同病变程度对移植效果的影响。方法:1、研究对象于2015年1月至2015年12月在我院行公民死亡后器官捐献肾移植术患者,并且术前供肾行零点活检,符合以下标准则纳入本研究:(1)、供肾组织病理检查每张切片必须同时包含肾小球、肾小管、肾间质及肾小血管四部分;(2)为了充分评估供肾组织病变,每张切片肾小球数量必须大于25个;(3)受者至少随访6个月以上;(4)供受者均为成人,受者为成人单肾移植;2、研究方法获取的供肾组织按照常规方法进行固定、脱水、石蜡包埋、切片及Masson's染色,基于肾脏病理Masson's染色形态学特征,采用Remuzzi评分系统对供肾组织的肾小球硬化、肾小管萎缩、肾血管损伤及肾间质纤维化病变程度进行评分,各个部分得分相加得出整个肾脏的总体得分。3、实验分组根据供肾病理的总得分分为两组,评分≤3为低分组,≥4为高分组.4、观察指标根据两组供肾肾移植受者进行术后随访1年,分析比较两组供肾肾移植术后移植肾功能延迟恢复(delayed graft function,DGF)、DGF恢复时长、术后3、6、12 个月肾小球率过滤(estimated glomerular filtration rate,eGFR)等相关指标的差异。5、统计方法统计分析采用SPSS 20.0统计软件。资料统计量采用均数±标准差(x±s)及百分比,组间均值差异性比较使用两独立样本t检验,组间率的差异性比较使用χ2检验或Fisher确切概率法,P0.05为差异有统计学意义。结果:供肾及相应受者共62例纳入本研究,供肾病变分值范围为0-5分,低分组44例,高分组18例,总体DGF为30.6%,两组间供肾对应供者年龄、平均住院时长及受者术后DGF发生率比较有显著性差异(P0.05),术后DGF恢复时长及1年内肾小球率过滤两组间比较均无统计学意义(P0.05)。结论:供肾组织病变程度越重,受者肾移植术后DGF发生率越高,平均住院时间越长。第二章非线性光学显微成像技术在供肾质量评估的应用目的:探究二次谐波结合双光子激发荧光的非线性光学显微成像技术在供肾质量评估。方法:研究对象: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献供肾零点活检组织。实验方法:零点活检组织标本经固定、石蜡包埋后,每个标本连续性切片两张,每张5um,一张行传统方法马松三色染色,每例切片在高倍镜下(200×)随机观察10个互不重叠肾小管间质视野并拍照,另一张行二次谐波结合双光子激发荧光显微成像,采用Remuzzi评分系统分别对供肾组织四种组件进行形态学半定量评分,即小管萎缩、间质纤维化、肾小球硬化、血管损伤及供肾整体得分,用K系数描述SHG/TPEF方法对各个组件和总体评分与传统Masson染色评估效果的一致性情况。统计方法:采用SPSS 20.0统计软件进行统计分析,用McNemar-Bowker检验Masson染色和SHG/TPEF两种方法算出的供肾组织各组件损伤评分构成比的差异,以PO.05为有统计学差异。一致性检验采用Kappa检验,当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。结果:非线性光学对供肾活检组织有高分辨率的组织细胞成像,能对供肾组织质量评估的肾小球、肾小管及血管有较好的形态学成像。具有不需要经过染色、标记,操作步骤简单、耗时短特点,采用Remuzzi评分系统分别对供肾组织小管萎缩、间质纤维化、肾小球硬化、血管损伤及供肾整体的半定量评分与Masson染色相比,差异无统计学意义(p0.05),在评估效果上与Masson染色评估效果一致性系数:肾小球硬化k=0.895,间质纤维化k=0.859,肾小管萎缩k=0.819,血管损伤k=0.802,总体情况k=0.914,全部一致性K系数大于0.75。结论:1、非线性光学显微成像技术在供肾质量评估效果与Masson染色评估效果相当;2、操作步骤简单、耗时短,具有运用于移植前供肾质量评估价值;3、对肾脏组织形态学上有较高分辨率。
[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.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R699.2
本文编号：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.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R699.2
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