MSX1、MSX2基因突变与眼—耳—脊柱发育不良关系的研究及孕期危险因素分析

发布时间：2018-01-21 14:44

本文关键词： 眼-耳-脊柱发育不良小耳畸形 MSX1 MSX2 孕期危险因素　出处：《复旦大学》2010年硕士论文　论文类型：学位论文

【摘要】： 目的本研究通过文献回顾,比较我院眼-耳-脊柱发育不良(oculo-auriculo-vertebral spectrum, OAVS)病例与国外病例临床特征的相同点与不同点；应用单因素卡方分析和logistic多因素回归分析的方法比较患者组与对照组母亲孕期情况的差异,以找到与OAVS相关的孕期高危因素,为产前预防提供思路；本研究应用PCR扩增目的DNA序列并直接测序的方法对OAVS患者血基因组MSX1、MSX2基因外显子突变进行检测,以了解该畸形与这两个基因外显子突变的关系,为进一步研究发病机制确定方向。方法本文收集了从2007.9到2009.7到我院门诊咨询或住院进行耳廓再造及听力重建术的208名眼-耳-脊柱发育不良患者。入选标准：患者有单／双侧耳廓畸形或附耳伴同侧半面短小；双侧受累定义为耳廓畸形、附耳或耳前瘘管在双侧出现；亲属中有耳廓畸形、附耳或瘘管的患者被定义为有家族史,其余为散发病例。根据国内外对先天性耳廓畸形和OAVS的相关研究,本研究制定了详细的《临床资料表》和《孕期危险因素调查表》。对于208例患者的临床资料,本研究与国外16篇文献比较了OAVS十项主要临床特征的百分比。208例患者中,有179例患儿母亲完成了《孕期危险因素调查表》,本研究选取患儿年龄在3-14岁的病例118例,并同时调查了118例3-14岁正常儿童母亲孕期情况作为对照组,使用SPSS 13.0统计软件,用卡方分析法对高龄产妇(35岁)、自然流产史、早孕感染、早孕用药史、早孕先兆流产史、早孕毒物接触史等16个变量进行了单因素卡方分析,挑选出有意义的因素后,再用logistic多因素回归法进行分析。此外,应用收集的124例患者血样,抽提出DNA后用PCR扩增了MSX1、MSX2基因外显子序列并直接测序,用Mutation Surveyor软件分析测序结果。结果 1.208例患者年龄在3月到31岁之间。69.7%的患者为男性；10.1%的患者有家族史,其余为散发病例；65.4%的患者单侧受累,单侧中56.6%为右侧受累；98.6%的患者伴发外耳道闭锁或狭窄；仅有14.4%的患者伴发附耳或耳前瘘管；28.4%的病例伴发耳畸形同侧或双侧下颌骨发育不良；4.3%的病例有不同眼部缺陷；只有2例患者确诊脊柱畸形,1例有先天性室间隔缺损。除了伴发外耳道闭锁或狭窄百分比明显高于国外报道,大部分结果与国外报道相似。 2.患者组和对照组在早孕接触有毒物质、孕期贫血、早孕感染、早孕用药史这4个方面差异有统计学意义(P0.05),可能为OAVS的孕期危险因素。其中早孕接触有毒物质的OR值最大,为9.847,说明母亲孕期接触油漆、化工原料等与OAVS的相关性很大。 3.在124例患者当中,发现1例患者MSX1基因第二个外显子上出现3473GGT突变,为无义突变,编码氨基酸仍为亮氨酸,此突变在100例正常人中验证均未发现异常。本研究未发现MSX2基因外显子突变,但发现MSXl,MSX2基因6个已报道过的SNP。结论本组OAVS患者临床特征与国外病例相似。患者母亲孕期贫血、早孕感染、早孕用药和早孕接触有毒物质等环境因素可能与发病相关。此先天性畸形由MSX1,MSX2基因外显子突变直接引起的可能性能不大。然而,有典型家族史的病例应该存在遗传学上的变异,需进一步进行分子生物学研究。
[Abstract]:The purpose of this study through the literature review, comparison of our hospital oculo ariculo vertebral dysplasia (oculo-auriculo-vertebral spectrum OAVS) common clinical features in the patients with foreign cases and different points; difference method using univariate chi square analysis and logistic regression analysis between patient group and control group pregnant condition, high risk pregnancy in order to find the factors associated with OAVS, to provide ideas for prenatal prevention; amplified target DNA sequence and direct sequencing of the research on the application of PCR in blood of patients with OAVS genomic MSX1, MSX2 gene exon mutations were detected, in order to understand the deformity and the two exon mutation relationship, determine the direction for the study of pathogenesis the mechanism further.
Methods from 2007.9 to 2009.7 in our hospital outpatient consultation or hospitalization for auricle reconstruction and reconstruction of hearing in 208 eyes - ears - spine dysplasia patients. Inclusion criteria: Patients with single / bilateral ear malformation or ear with ipsilateral hemifacial microsomia; bilateral involvement is defined as the auricle malformation, ear or ear fistula in bilateral relatives; auricle deformity, ear or fistula patients were defined as having family history, the rest were sporadic. According to the related research on congenital auricular deformities and OAVS at home and abroad, this study developed a detailed table > and < < clinical pregnancy risk factors for the clinical data of 208 questionnaires. Patients in this study, with 16 foreign literatures and comparison of the OAVS ten main clinical features of the percentage of.208 patients, 179 patients completed the "mother's pregnancy risk factors questionnaire", this study selected children in At the age of 3-14 years old in 118 cases, and also investigated the 118 cases of 3-14 years old children with normal pregnancy as control group, using SPSS 13 statistical software, advanced maternal age on using chi square analysis (35 years old), the history of spontaneous abortion in early pregnancy, infection, pregnancy medication history, pregnancy history of early pregnancy threatened abortion, poison contact history 16 variables are single factor chi square analysis, select significant factors, analysis and multiple factor Logistic regression method. In addition, 124 cases of patients with blood samples collected using, extracted after DNA was amplified by PCR MSX1 and MSX2 gene exon sequences and direct sequencing, sequencing and analysis of results using Mutation Surveyor software.
Result
1.208 patients at the age of.69.7% from March to 31 between the ages of the patients were male; 10.1% of patients have family history, for the rest of 65.4% sporadic cases; patients with unilateral involvement, 56.6% unilateral right hair involvement; external auditory canal atresia or stenosis and 98.6% patients with ear hair or preauricular fistula; only 14.4% of the patients; 28.4% of the patients with microtia ipsilateral or bilateral mandibular dysplasia; 4.3% cases with different eye defects; only 2 cases were diagnosed as spinal deformity, 1 cases with congenital ventricular septal defect. In addition to aural atresia or stenosis was significantly higher than that reported abroad, most similar to the results reported abroad.
2. patients group and control group exposed to the toxic substances in early pregnancy anemia during pregnancy, early pregnancy infection, there were statistically significant differences in early pregnancy medication history of these 4 aspects (P0.05), may be the risk factors during pregnancy OAVS. The early exposure to toxic substances and the OR value is 9.847, that maternal exposure to chemical raw materials such as paint, correlation with OAVS a lot.
3. of the 124 patients, 1 patients found that second of the MSX1 gene exon 3473GGT mutation, nonsense mutation, encoding amino acids are leucine, this mutation in 100 cases of normal people in verification were not unusual. This study found no mutations of exons of MSX2 gene, but the MSXl. 6 MSX2 genes have been reported SNP.
Conclusion the clinical characteristic of OAVS patients with foreign similar cases. Patients with maternal anemia during pregnancy, early pregnancy infection, pregnancy and early pregnancy medication exposure to environmental factors of toxic and hazardous substances may be correlated with the incidence of congenital malformation. This consists of MSX1, MSX2 gene mutation in exon may be directly caused by the performance little. However, some typical cases of family history should be there is variation in genetics, the further research of molecular biology.

【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R764

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