本文選題：激素耐藥型腎病綜合征 + 單卵雙胞胎��； 參考：《福建醫(yī)科大學(xué)》2012年碩士論文

【摘要】：第一部分激素耐藥型腎病綜合征單卵孿生姐妹WT1基因突變分析 目的分析1個南方漢族激素耐藥型腎病綜合征（SRNS）家系WT1基因突變及其特點。方法研究對象為1個南方漢族SRNS家系先證者及其單卵孿生姐姐、父母和大姐，對照人群為50例尿檢正常的漢族成年人。取所有研究對象外周靜脈血3mL，提取基因組DNA，PCR擴(kuò)增WT1基因全部10個外顯子及其周圍的部分內(nèi)含子序列，對PCR產(chǎn)物直接進(jìn)行DNA序列測定。結(jié)果先證者臨床表型為不完全型Denys-Drash綜合征，其孿生姐姐臨床表型為孤立性SRNS。在該對單卵孿生姐妹均檢測出WT1基因1180CT（R394W）雜合突變，在先證者父母及其大姐和50例對照人群未檢測出1180CT（R394W）雜合突變；此外，還在該對孿生姐妹檢測出3個相同的WT1基因多態(tài)性—126CT、903AG和IVS7 32CA。結(jié)論本研究在1個漢族SRNS家系臨床表型不同的單卵孿生姐妹患兒發(fā)現(xiàn)了相同的WT1基因R394W雜合突變。 第二部分中國漢族散發(fā)性孤立性激素耐藥型腎病綜合征女性患兒WT1基因突變分析 目的分析14例漢族散發(fā)性孤立性激素耐藥型腎病綜合征（ISRNS）女性患兒WT1基因突變及其特點。方法研究對象為14例漢族散發(fā)性ISRNS女性患兒（已除外NPHS2基因突變），對照人群為38例漢族散發(fā)性ISRNS男孩和50例尿檢正常的漢族成年人。取所有研究對象外周靜脈血3mL，提取基因組DNA，PCR擴(kuò)增WT1基因外顯子8和9及其周圍的部分內(nèi)含子序列，對PCR產(chǎn)物直接進(jìn)行DNA序列測定。結(jié)果對14例漢族散發(fā)性ISRNS女性患兒進(jìn)行WT1基因突變檢測，結(jié)果在1例散發(fā)性ISRNS女孩患兒檢測出WT1基因IVS9+5GA雜合變異，該變異在50例正常對照人群中未檢測出。漢族散發(fā)性ISRNS女性患兒WT1基因突變檢出率7.1%。結(jié)論中國漢族散發(fā)性ISRNS女性患兒存在WT1基因突變，提示對漢族散發(fā)性ISRNS女性患兒需進(jìn)行WT1基因突變分析。 第三部分1例漢族孤立性腎病綜合征伴腎衰竭男性患兒WT1基因突變分析 目的分析1例漢族孤立性腎病綜合征伴腎衰竭男性患兒WT1基因突變及其特點。方法研究對象為1例漢族孤立性腎病綜合征伴腎衰竭男性患兒，6.3歲發(fā)病，，對照人群為110例尿檢正常的漢族成年人。取所有研究對象外周靜脈血3mL，提取基因組DNA，PCR擴(kuò)增WT1基因全部10個外顯子及其周圍的部分內(nèi)含子序列，對PCR產(chǎn)物直接進(jìn)行DNA序列測定。結(jié)果在該患兒WT1基因外顯子8檢測出1051AG雜合變異，但在110例正常對照人群中未檢測出1051AG變異，表明該變異為WT1基因突變。該突變導(dǎo)致WT1基因的編碼蛋白WT1的第351位賴氨酸被谷氨酸取代，即K351E。經(jīng)檢索PUBMED文獻(xiàn)數(shù)據(jù)庫、單核苷酸多態(tài)性數(shù)據(jù)庫和人類基因突變數(shù)據(jù)庫已公布的WT1基因變異，證實1051AG（K351E）為新發(fā)現(xiàn)的WT1基因突變。結(jié)論在1例6.3歲孤立性腎病綜合征伴腎衰竭男性患兒檢測出WT1基因突變，提示對孤立性腎病綜合征伴腎衰竭男性患兒需進(jìn)行WT1基因突變分析。
[Abstract]:Analysis of WT1 gene mutation in monozygotic twin sisters of steroid-resistant nephrotic syndrome Objective to analyze the mutation and characteristics of WT1 gene in a Chinese family with steroid-resistant nephrotic syndrome. Methods the subjects of this study were a proband from a SRNS family of southern Han nationality and their monozygotic twin sisters, parents and elder sisters, and a control group of 50 Han adults with normal urine tests. All 10 exons of WT1 gene and some intron sequences around them were amplified by genomic DNA-polymerase chain reaction (DNA-PCR) from peripheral venous blood of all subjects. The PCR products were sequenced by DNA directly. Results the clinical phenotype of the proband was incomplete Denys-Drash syndrome, and the twin sister's clinical phenotype was isolated. Heterozygous mutations of WT1 gene were detected in all of the single twin sisters, and no heterozygous mutations were detected in the proband parents and their elder sisters and 50 controls. Three identical WT1 gene polymorphisms-126 CTG 903AG and IVS7 32 CA were also detected in the twin sisters. Conclusion the same R394W heterozygous mutation of WT1 gene was found in a single twin sister with different phenotypic phenotypes in a SRNS pedigree of Han nationality. The second part of the Chinese Han nationality sporadic isolated hormone resistant nephrotic syndrome in women with WT1 gene mutation analysis Objective to analyze the mutation and characteristics of WT1 gene in 14 children with sporadic isolated steroid-resistant nephrotic syndrome (ISR NSS) of Han nationality. Methods the subjects were 14 sporadic ISRNS female children (excluding NPHS2 gene mutation), 38 Han sporadic ISRNS boys and 50 normal controls. The exons 8 and 9 of WT1 gene and some intron sequences around them were amplified by PCR from peripheral venous blood of all subjects. The PCR products were sequenced by DNA directly. Results the mutation of WT1 gene was detected in 14 sporadic ISRNS female children of Han nationality. The results showed that the IVS9 5GA heterozygosity of WT1 gene was detected in 1 sporadic ISRNS girl, but not in 50 normal controls. The detection rate of WT1 gene mutation in sporadic ISRNS women of Han nationality was 7.1%. Conclusion there is a mutation of WT1 gene in sporadic ISRNS female children of Han nationality in China, suggesting that WT1 gene mutation analysis should be carried out in sporadic ISRNS female children of Han nationality. Part three: analysis of WT1 gene mutation in children with isolated nephrotic syndrome with renal failure in Han nationality Objective to analyze the WT1 gene mutation and its characteristics in a patient with isolated nephrotic syndrome with renal failure in Han nationality. Methods one patient with isolated nephrotic syndrome associated with renal failure in Han nationality aged 6.3 years was studied, and 110 Han adults with normal urine test were enrolled in the study. All 10 exons of WT1 gene and some intron sequences around them were amplified by genomic DNA-polymerase chain reaction (DNA-PCR) from peripheral venous blood of all subjects. The PCR products were sequenced by DNA directly. Results 1051AG heterozygosity was detected in exon 8 of WT1 gene, but no 1051AG mutation was detected in 110 normal controls, indicating that the mutation was a mutation of WT1 gene. The mutation resulted in the substitution of lysine at position 351 of WT1, the encoding protein of WT1 gene, by glutamate, K351E. By searching PUBMED literature database, single nucleotide polymorphism database and human gene mutation database, it was confirmed that 1051 AGN K351E was a new WT1 gene mutation. Conclusion WT1 gene mutation was detected in a 6.3 year old boy with solitary nephrotic syndrome with renal failure, suggesting that WT1 gene mutation should be analyzed in the isolated nephrotic syndrome with renal failure.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R726.9

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