中国重庆AIS遗传流行病学调查和SH3GL1序列比对分析研究
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摘要
研究背景及研究目的:
青少年特发性脊柱侧弯(Adolescent Idiopathic Scoliosis简称AIS)是一种伴有椎体旋转的脊柱结构性侧向弯曲的三维畸形。在儿童和青少年中的发病率国外报道为2%~5%,国内报道为1.5%~3%,重度畸形的发病率为0.2~0.5%,女性患者明显多于男性。虽然AIS除表现畸形外通常不引起严重的病理状态,但是重度胸弯可能引起心、肺功能的减退,导致明显的体型上的畸形,甚至影响寿命。目前治疗方法主要是针对畸形进行矫治,包括临床观察、支具治疗和对侧弯超过45o者进行手术治疗,治疗复杂,常遗留有脊柱畸形和脊柱活动障碍。效果不理想,理想的效果是消灭畸形。如果能够针对病因进行症状前诊治、出生前诊治,甚至对有家族史的已孕女性,在受精6天、胚胎着床之前进行基因诊断和治疗,杜绝脊柱畸形的出现,临床上将消灭AIS。但这需要首先明确病因,明确发病的基因机制。
目前AIS的确切病因尚不清楚。既往大量的研究集中在软骨、骨骼、神经和肌肉等组织学因素的致病机制上面,但是没有明确结论。随着医学遗传学和医学分子生物学研究的不断深入和进步,临床上许多原因不明的常见病的发病机制被证明与遗传及致病基因有关。目前医学分子生物学的观点认为,人体各种疾病的发生,都是与一种或多种基因有关。这样人们自然想到AIS是否与遗传有关,近来许多学者认为AIS发病与遗传有关,如果与遗传有关,就需要确定它的遗传模(单基因遗传模式还是多基因遗传模式)和疾病基因的位置和疾病的分子机制,探讨遗传因素对疾病的影响和可能的遗传模式是遗传流行病学的任务。探讨疾病基因的位置和引起疾病的机制是医学分子生物学研究任务。
AIS流行病学方面和遗传流行病学方面的研究报道很少,但这些研究都报道具有家族聚集性,认为有遗传因素,至于遗传模式,各家报道的结果不同一,有报道认为是常染色体显性遗传模式,有报道是X连锁显性遗传模式,还有报道是多基因遗传模式。也就是说AIS是不是遗传病,遗传模式是哪种还没有定论。没有确定的遗传模式,这对AIS疾病基因的染色体定位研究带来了障碍,进一步影响疾病基因的筛查与鉴定。在遗传病的疾病基因的染色体定位分析过程中,需要根据不同的遗传模式而采用不同的分析方法,一般单基因病采用连锁分析方法,多基因病采用相关分析法。如果分析方法反了或混合了,结果不言而逾,也就是说在遗传模式不确定的情况下,其分析的结果是不确定的,这也正是为什么现有的、关于基因染色体定位研究报道结果各异,无法统一的原因之一(亦有遗传异质性问题)。所以确定AIS是遗传病,是那种模式的遗传病的研究,也就是遗传流行病学方面的研究,对疾病基因的定位和基因机制的研究具有重要意义。
AIS的医学分子生物学机制的研究已有部分报道,各学者从“正向”或“反向”两种策略进行研究,但目前仍处于疾病基因位点的早期染色体定位研究阶段,并且尚没有公认的同一的基因位点。“正向”研究策略(经典遗传学策略)既功能克隆法是从蛋白质或mRNA表达差异着手,采用以消减杂交为思路的多种分子生物学手段,先通过消减获得特异表达和缺失的基因片段,然后进行染色体定位乃至获得全基因。关于AIS病变部位不同组织的mRNA和蛋白质表达差异方面的研究已有许多报道,研究集中在对侧弯脊柱凸凹侧椎间盘内的Ⅰ、Ⅱ、X型胶原,TGF-?1、bFGF、成骨细胞蛋白质表达差异和ⅠⅡ、Ⅸ型胶原mRNA的表达差异方面,虽然均报道存在表达差异,但和组织学研究结果一样,这种差异“是因是果”难以确定,无法确定真正的病因。这种现状的原因是研究设计都是采用凸、凹侧对比,如果想找出真正的组织、蛋白或mRNA层面的差异,需要进行先证者自身发病前后对比研究,或先证者与正常人之间的对比研究,但是这在临床上是很难做到的,所以结果很难取得突破,那么只有寄希望于“反向”研究了。
“反向遗传学”策略既医学分子生物学策略是应用定位克隆遗传分析法对疾病基因进行染色体定位和疾病基因的筛查与鉴定。这种方法是基于任何一个基因必定在染色体上有位点,基因所决定的遗传学性状与这些位点有密切联系的遗传学原理,通过遗传学分析,获得某些疾病基因在染色体上的位置信息,然后根据这一位置信息,应用“物理图”的物理标记将遗传学信息转变为“物理标记”所代表的明确的染色体区域,再以相关区域的“相连片段群”来筛选可表达的基因,建立该区域的“转录图”。最后,通过“定位候选克隆”和“功能候选克隆”方法比较病人与正常人这些结构基因的区别以确定与疾病相关的改变,筛查与鉴定出疾病相关基因。关于AIS疾病基因的定位克隆研究共检索到7篇文献,由于各个研究应用了不同的研究群体和遗传统计学分析方法,各家报道的染色体定位的位点各异,分别在染色体6、9、10q、16、17、17p11、19p13.3、19 p 13和Xq21上,还没有取得公认的定位结果。
值得关注的是在定位克隆法研究中,有2位学者将疾病基因位点定在19p13 ,Alden研究的受试对象是美国人。Chan的报道不仅定位在19p13.3内,还进一步的精确定位于D19S216标志附近,在D19S894～D19S1034之间的5.2cM遗传距离,物理图约1.9Mb。研究的受试对象是中国广东人,采用400多个荧光标记的微卫星标志进行全基因组筛查,选用了参数和非参数两种相关分析的统计方法,基本排除了遗传模式对结果的影响,从一个家系中得到的结果在另外6个家系中得到验证,样本量大。文章的影响因子高达10.0以上,结果可信。此项研究虽然精确的定位到D19S216标志附近,但仍然属于疾病基因的染色体定位研究,未见后续的关于疾病基因的筛查与鉴定的研究报道。
我们认为承认Chan的研究结果,在其研究基础上做进一步的研究——候选基因筛查,以期在AIS的疾病基因定位研究上取得突破性进展是值得的,是有创新的和有科学价值的。为了确定19p13.3内基因的数目,检索在3亚带内有198个已知基因,其中只有17个是已知疾病的基因,尚无与AIS相关的基因报道。
如果要对3亚带内的198个基因进行筛查,从哪个基因开始是值得研究的。当然是选与AIS最相关的,Chan的精确区域定位是在D19S216标志附近,在物理图上查SH3GL1恰位于此区域内, Chan在报道中也指出SH3GL1是与破骨细胞代谢有关,可能是AIS重要的致病基因。
首选SH3GL1。关于基因SH3GL1功能的研究报道不多,Wise等人报道基因SH3GL1与细胞吞噬作用有关,参与细胞内吞、外排偶联的Ca2+通道的调节。也有研究认为SH3GL1与人类的白血病有关。
所以本项研究的目的一是对中国重庆AIS进行遗传流行病学调查分析,探讨遗传因素在发病中的作用和可能的遗传模式。二是采取定位候选克隆方法对AIS的疾病候选基因SH3GL1的外显子进行序列比对分析,研究是否存在碱基变异,为疾病基因筛查与鉴定做方法学准备。
研究方法:
1.遗传流行病学调查:设计AIS遗传流行病学调查表,收集在西南医院诊治过的先证者资料。确立诊断标准为站立前后位X线片Cobb角>10o为受累者, Cobb角<10o为易感者,年龄小于12岁的表型正常者为未确定者。由经过培训的调查员对先证者及其家系成员进行询问调查并填写调查表。方法为门诊、病房询问与电话询问相结合,为防止存在回忆偏倚对先证者、父亲和母亲分别进行询问记录。对怀疑受累者和易感者要进行临床望诊、Adam弯腰试验检查,必要时经过X线片检查确定。
2.遗传流行病学统计分析:对遗传流行病学调查数据采用SPSS10.0统计软件进行分析。通过年龄发病率分析,家族聚集性分析,遗传度计算(Falconer法)分析确定遗传因素的大小。根据单基因和多基因遗传病的特点分析确定遗传模式。
3.定位候选克隆:设计以家系为基础的“病例同胞对”研究方案,采集“同胞对”的血液标本,提取基因组DNA,根据基因SH3GL1核酸序列,以10个外显子为重点设计引物对,进行PCR扩增、克隆和测序。
4.序列比对分析:用GeneTool软件对外显子测序结果,进行“同胞对”之间和同胞对与NCBI之间的序列比对分析,判断是否存在碱基变异,并进行蛋白质结构预测分析。
研究结果:
1.遗传流行病学调查:设计了AIS遗传流行病学调查表,并成功的对73个家系按调查表进行遗传流行病学调查。
2.遗传流行病学统计:年龄发病率分析:发病75%在11～14岁之间,平均年龄10.84岁。家族聚集性分析:病例家族总发病率为4.86%,一级亲属的发病率为16.93%,二级亲属的发病率为7.21%,三级亲属发病率4.36%,而参照发病率为2.25%,统计学分析差异有显著意义(u=6.451,P﹤0.01 )。遗传度计算:先证者一级亲属的遗传度84%、二级亲属的遗传度88%和三级亲属的遗传度85%,经显著性检验(P<0.01)各级亲属遗传度有统计学意义。
3.定位候选克隆:候选基因SH3GL1中的10个外显子成功的在AIS“同胞对”的基因组DNA中得到扩增和克隆,并且序列测定均取得阳性结果。
4.序列比对分析:通过比对分析在候选基因SH3GL1的10个外显子中,共发现12处碱基变异,分别位于第2(2处)、4、5(3处)、6、8和10(2处,非编码区)六个外显子上。其中先证者基因编码mRNA第515位碱基如果为T,则形成终止密码子(TAG),导致蛋白阅读框发生改变(可能为107-518区域碱基,或为707-1213碱基),蛋白质序列预测分析显示编码截短蛋白,从而影响蛋白质的一级结构。
结论:
1.AIS是遗传病:年龄发病率分析显示75%在11～14岁之间,比较集中,属于遗传病的特点;病例家族发病率和先证者一级亲属、二级亲属和三级亲属的发病率均明显高于参照群体平均发病率,具有明显的家族聚集性;一级、二级和三级亲属的遗传度均大于84%,证明AIS与遗传显著相关。综合上述分析结果证明AIS是遗传病。
2.AIS是多基因遗传病:先证者的各级亲属发病率均明显高于参照人群发病率;同胞发病率远比1/2或1低;与先证者的亲缘关系越近,发病率越高,有明显的家族性聚集倾向。综合分析证明AIS属于多基因遗传病。
3.基因SH3GL1是AIS的致病基因之一:碱基序列比对分析证明外显子区域内的10处碱基变异位于编码区属于结构基因的变异,2处位于非编码区内的变异可能处于调控区域,先证者SH3GL1第5外显子的第515位碱基如果发生C→T突变,将形成终止密码子,导致蛋白阅读框发生改变,通过氨基酸序列预测分析,编码截短蛋白,从而显著影响蛋白质的结构。由此可以证明基因SH3GL1是AIS候选基因之一。
Background and Objects of the Study
Adolescent Idiopathic Scoliosis (AIS) is a three-dimensional deformation of dorsal spine structural perversion with vertebral body rotation. The incidence of children and teenagers is 2%~5% from reports abroad while incidence is 1.5%~3% from domestic reports. The incidence of severe deformation is 0.2~0.5% with female patients obviously more than male ones. Though ALS ususlly does not cause severe pathologic state except deformation, severe thoracocyrtosis may cause heart and lung hypofunction, leading to conspicuous somatotype deformation and even life. At present, the available treatments focus on correcting and curing on deformation including clinical observation, orthosis treatment, and operation on patients with lateral curvature above 45o. However, the treatment process was complicated with spinal deformity and dorsal spine activity obstacle left, producing no ideal effect. The ideal effect is to eliminate deformity. If diagnosis and cure on etiological factor can be made before symptoms and birth, or even gene diagnosis and cure can be made for pregnant women with family medical history before 6 days of ferrtilization and embryo nidation, spinal deformity will be cut off and AIS eliminated clinically. But etiological factor and invasive gene mechanism need to be identified first.
So far, The definite etiological factor of AIS has not been detected. Past researches concentrated on pathogenic mechanism of histology factors such as cartilage, ossature, nerves and muscle, but no definite conclusions were drawn. With unceasing deepening and progress of researches on medical genetics and medical molecular biology, clinically pathogenesy of many agnogenic common diseases has been proved relevant with heridity and virulence gene. At present, medical molecular biology holds that generation of various kinds of diseases is related with one or many genes. Thus people are easy to think that whether AIS is concerned with heredity? Recently many scholars deem AIS correlates with heredity. If it is related with heredity, what is the heredity mode? monogenic inheritance mode or polygenic inheritance mode? How is the genic location and molecule mechanism of disease? The approach to influence of genetic factors on disease and possible hereditary modes is the task of genetic epidemiology, while the approach to disease genic location and mechanism evoking disease, the research task of medical molecular biology.
There are few research reports about AIS epidemiology and genetic epidemiology, but these reports show familial aggregation. Genetic factor was deemed to be involved with AIS, which has several inheritance modes including autosomal dominant inheritance mode, X-linked dominant inheritance mode, and polygenic inheritance mode. That is to say, no final conclusion is drawn about whether AIS is genetic disorder and what kind of hereditary mode it is. No definit hereditary mode bring obstacle for chromosomal assignment study on AIS pathological gene The screening and identification of pathological gene were also affected consequently. Different analytical methods were applied in accordance with corresponding inheritance mode during the process of Chromosomal assignment analysis on pathological gene. Linkage analytical method was applied for monogenic disorder while correlation analysis for polygenic disease. The analytical results were indefinite with indefinite inheritance mode, which gave rise to different research results of Chromosomal assignment study. The decisive role was played by discussions on the influence of genetic factor on AIS and the possible inheritance mode in Chromosomal assignment study on AIS pathological gene as well as screening and identification of pathological gene. Only by determining AIS genetic factor and inheritance mode can Chromosomal assignment of pathological gene be conducted by employing linkage analysis (analytical method aiming at monogenic inheritance disorder) or correlation analysis (analytical method aiming at polygenic inheritance disorder) under the guidance of the inheritance mode.
Some studies on AIS molecular genetics mechanism have been reported. Two strategies (Norientation or uorientation ) were employed to the study. However the study is still at the stage of early chromosomal assignment study of the disease gene locus, and up to now a consensus gene locus has not been determined. Norientation strategy (classical genetics), namely the cloning methods based on gene function, was oriented to function of proteic substance or differential gene expression of mRNA. Multiple molecular biology methods oriented to differential hybridization were applied. Initially specific expression and the lacked gene fragment were acquired by subtraction, and then chromosomal assignment ending with obtaining whole gene was conducted. Studies on mRNA or differential expression of proteic substance of different tissues of AIS diseased region have been reported. They focused onⅠ、Ⅱ、X type collagen, TGF-?1、bFGF and osteoblast protein expression differences inside intervertebral cartilages in lateral curvature dorsal spine protruding and curved side andⅠ、Ⅱ、Ⅸtype collagen mRNA expression difference. Although it has been unanimously reported that differential expression existed, yet it has not been determined whether the differential expression was etiological factor or the result of disease. That is to say, the genuine etiological factor has not been detected. The reason of this status quo is that research and design use contrast of protruding and curved side. If differences of genuine tissue, protein or mRNA are needed to be found, contrast research of proband before and after invasion or research between proband and normal people should be made. But it is hard to make clinically, so the result is hard to break through with the only hope for“reversing”research.
Positional cloning genetic analysis was applied to the chromosomal assignment, screening, and identification of disease gene in the strategy of reverse genetics (molecular genetics). The method was premised on the genetics principles that any gene has its gene locus in chromatin body, and genetics characters determined by genes were closely correlated with these gene loci. position information of disease gene in chromatin body was obtained through genetics analysis, then according to the position information, the physical marking of“physical map”was applied to the transformation from genetics information to definite chromatin region represented by the physical marking, then the expressible gene was screened in accordance with linked gene fragment grouping of the correlated regions, and transcription map of the region could be made. Finally two methods of positioning candidate cloning and function candidate cloning were employed to compare the differences of architectural gene between the patients and the healthy one, thus changes correlated with the disease were defined, and disease gene was screened and identificated. 7 reports about positional cloning study on AIS disease gene has been retrived, yet a consensus conclusion has not been drawn due to different study groups, genetics statistical analytical method, and gene loci of chromosomal assignment. No received positioning results were got respectively for chromosome 6、9、10q、16、17、17p11、19p13.3、19 p 13 and Xq21.
It should be noticed that two scholars positioned the disease gene locus in 19p13 in their positional cloning studies. Their trail objects were Americans and Cantonese respectively. The key-gene locus of the latter was 19p13.3, and minor gene locus was 2p, which were approved to be located in the vicinity of the mark of D19S216 , ranging in the breadth of 5.2cM between D19S894～D19S1034 with the approximate length of 1.9Mb through further accurate positioning and recombination. Research objects that receive experiment are Cantonese. More than 400 fluorescently-labeled microsatellite signals were applied to make the whole genome screening, and two correlation analysis statistical methods of parameter and nonparameter were adopted. Influence of hereditary mode on results was basically eliminated. Besides, results from one family constellation were checked in other 6 family constellations with large sample size. Affectoi of article is as high as over 10.0, producing a convincing result. Although the disease gene locus was accurately positioned in the vicinity of the mark of D19S216 in the study, the research was still in the province of chromosomal assignment study of disease gene. The following screening and identification of pathological gene were not reported, which adequately approved that the gene locus was highly likely to correlate with AIS. Methods of positioning candidate cloning were worthy applying to the study on screening and identification of 198 genes in the position area.
To make sure the number of intragenes 19p13.3, 198 known genes inside 3 subband are retrived, among which only 17 are genes of known diseases. Gene reports related with AIS has not been attained.
If 198 genes inside 3 subband are to be screened, which gene to be start with were worthy studie. Certainly, gene that is the most related with AIS is chosen. Accurate positioning of Chan is in the vicinity of the mark of D19S216. From physical map, SH3GL1 is just located in this area. In report, Chan also pointed that SH3GL1 is concerned with osteoclast metabolism, and may be the important virulence gene of AIS.
SH3GL1 is the first choice.There are not many research reports about function of gene SH3GL1. People like Wise reported that gene SH3GL1 is revelant with cytophagocytesis, and participated in adjustment of Ca ion channel of cell internalization and excretion couplant. Some researches also think that SH3GL1 is related with leukemia in human beings.
Therefore the study was designed to make genetic epidemiologic survey and analysis of AIS in Chongqing and discuss the effect of inheritance factors on the AIS and the possible inheritance mode. Moreover, the method of positioning candidate cloning was applied to the analytical study on the Exon Sequence Alignment of the AIS Disease Candidate Gene SH3GL1, making technology preparation for the screening and identification of the disease gene.
Research Method
1.Genetic epidemiologic survey: The genetic epidemiology questionnaire for AIS was designed, and the data of index case who received medical treatment in Southwest Hospital were collected. Diagnostic code: affected individual: std. A-P position X-ray Cobb Ang > 10o; susc.: Cobb Ang <10o; the undetermined : normal phaenotype and age under 12. Initially trained investigators inquired index cases and their family members, and then filled the questionnaires. Investigating methods include out-patient clinic, hospital room inquiry, and telephone inquiry. Index cases and their fathers and mothers were inquired respectively to avoid memory errors. The suspected affected individual and susc. were received clinical inspections, Adam waist-bending test, and X-ray check when necessary.
2. Statistical analysis of genetic epidemiology: SPSS10.0 statistics software was applied to the analysis of the genetic epidemiology survey data. The influence of genetic factor was determined by analyses on disease age incidence, familial aggregation, and calculation of degree of heritability. Inheritance mode was determined according to analysis of features of the disease of monogenic or multifactorial inheritance.
3. Positioning candidate cloning:“case-sibling control design”research scheme based on family constellation was designed. A sib pair was choosed to collect blood preparation, and genome DNA was extracted. According to nucleotide sequence of gene SH3GL1, primer pair for PCR amplification, cloning, and sequencing with 10 exons as emphasis was designed.
4. Sequence comparative analysis: sequence comparative analysis for exon sequencing result between sib pairs, and that between sib pair and NCBI were conducted through GeneTool software to judge whether basic group mutation occurred or not. amino acid sequence comparative analysis for predict was made.
Research Results
1. Genetic epidemiologic survey: The genetic epidemiology questionnaire for AIS was designed and genetic epidemiology survey on 73 family constellations was conducted successfully according to the questionnaire.
2. Statistical analysis of genetic epidemiology: analysis of disease age incidence: morbility of the disease of 75% occurred between 11and 14. The mean morbility age was 10.84. Analysis of familial aggregation: family total disease incidence of the case was 4.86%. Disease incidence of first degree relative was 16.93%, and disease incidence of second degree relative was7.21%, and disease incidence of third degree relative was 4.36%, and disease incidence of the reference family was 2.25%. The incidences were significant differences by statistics analysis. (u=6.451,P﹤0.01 )Calculation of degree of heritability (Falconer Method): degree of heritability of first degree relative was 84%, degree of heritability of second degree relative was 88%, and degree of heritability of third degree relativ was above 85%, which had statistical significance through significance test (P<0.01)
3. Positioning candidate cloning: 10 exons of the candidate gene SH3GL1 were successfully amplified and cloned in genome DNA of an AIS sib pair, and the sequencings obtained positive results.
4. Sequence comparative analysis: 12 basic group mutations were found in 10 exons of the candidate gene SH3GL1 of patients with AIS. These mutations were located in the second exon (2 mutations), the fourth exon, the fifth exon (3 mutations), the sixth exon, the eighth exon, and the tenth exon (2 mutations, noncoding region). If basic group in 515 of mRNA was mutationed to T, stop codon(TAG) was made and open reading frame was altered , the sequence of proteinum was predicted and show brachytmema proteinum was encoded which could cause changes of primary structure.
Conclusions:
1. AIS was genetic disorder. Morbility of the disease usually occurred between 11 and 14 according to the analysis of disease age incidence. The family disease incidence of the case, disease incidence of first degree relative and the second degree relative and third degree relative were found obviously higher than the average disease incidence of the reference group. AIS was found to have obvious familial aggregation. The degree of heritability of first degree relative and second degree relative and third degree relativ was above 84%, which approved AIS was significantly correlated with heredity. AIS belonged to genetic disorder through comprehensive analysis. 2.AIS was a disease of multifactorial inheritance. On the whole, AIS was a genetic disorder; and the disease incidences of the index case’s all degrees of relatives were found to be higher than that of the reference crowd. Sibling disease incidence was found far lower than 1/2 or 1. The closer blood relationship with the index case means higher disease incidence. The disease has obvious familial aggregation. Among 73 family constellations, only 3 of them were found to have the family history of the disease. Index cases in other family constellations were all single cases. AIS belonged to the category of a disease of multifactorial inheritance through comprehensive analysis.
3. Gene SH3GL1 was one of the virulence genes of AIS. sequence comparative analysis show that Basic group mutations were found in exons of the candidate gene SH3GL1, and 10 of the mutations were located in the coding region and 2 of the mutations were located in the noncoding region. If basic group in 515 of mRNA was mutationed from C to T, stop codon(TAG) was made and open reading frame was altered , the sequence of proteinum predict show brachytmema proteinum was encoded which could cause changes of primary structure. It was demonstrated that Gene SH3GL1 was one of the virulence genes of AIS.
青少年特发性脊柱侧弯(Adolescent Idiopathic Scoliosis简称AIS)是一种伴有椎体旋转的脊柱结构性侧向弯曲的三维畸形。在儿童和青少年中的发病率国外报道为2%~5%,国内报道为1.5%~3%,重度畸形的发病率为0.2~0.5%,女性患者明显多于男性。虽然AIS除表现畸形外通常不引起严重的病理状态,但是重度胸弯可能引起心、肺功能的减退,导致明显的体型上的畸形,甚至影响寿命。目前治疗方法主要是针对畸形进行矫治,包括临床观察、支具治疗和对侧弯超过45o者进行手术治疗,治疗复杂,常遗留有脊柱畸形和脊柱活动障碍。效果不理想,理想的效果是消灭畸形。如果能够针对病因进行症状前诊治、出生前诊治,甚至对有家族史的已孕女性,在受精6天、胚胎着床之前进行基因诊断和治疗,杜绝脊柱畸形的出现,临床上将消灭AIS。但这需要首先明确病因,明确发病的基因机制。
目前AIS的确切病因尚不清楚。既往大量的研究集中在软骨、骨骼、神经和肌肉等组织学因素的致病机制上面,但是没有明确结论。随着医学遗传学和医学分子生物学研究的不断深入和进步,临床上许多原因不明的常见病的发病机制被证明与遗传及致病基因有关。目前医学分子生物学的观点认为,人体各种疾病的发生,都是与一种或多种基因有关。这样人们自然想到AIS是否与遗传有关,近来许多学者认为AIS发病与遗传有关,如果与遗传有关,就需要确定它的遗传模(单基因遗传模式还是多基因遗传模式)和疾病基因的位置和疾病的分子机制,探讨遗传因素对疾病的影响和可能的遗传模式是遗传流行病学的任务。探讨疾病基因的位置和引起疾病的机制是医学分子生物学研究任务。
AIS流行病学方面和遗传流行病学方面的研究报道很少,但这些研究都报道具有家族聚集性,认为有遗传因素,至于遗传模式,各家报道的结果不同一,有报道认为是常染色体显性遗传模式,有报道是X连锁显性遗传模式,还有报道是多基因遗传模式。也就是说AIS是不是遗传病,遗传模式是哪种还没有定论。没有确定的遗传模式,这对AIS疾病基因的染色体定位研究带来了障碍,进一步影响疾病基因的筛查与鉴定。在遗传病的疾病基因的染色体定位分析过程中,需要根据不同的遗传模式而采用不同的分析方法,一般单基因病采用连锁分析方法,多基因病采用相关分析法。如果分析方法反了或混合了,结果不言而逾,也就是说在遗传模式不确定的情况下,其分析的结果是不确定的,这也正是为什么现有的、关于基因染色体定位研究报道结果各异,无法统一的原因之一(亦有遗传异质性问题)。所以确定AIS是遗传病,是那种模式的遗传病的研究,也就是遗传流行病学方面的研究,对疾病基因的定位和基因机制的研究具有重要意义。
AIS的医学分子生物学机制的研究已有部分报道,各学者从“正向”或“反向”两种策略进行研究,但目前仍处于疾病基因位点的早期染色体定位研究阶段,并且尚没有公认的同一的基因位点。“正向”研究策略(经典遗传学策略)既功能克隆法是从蛋白质或mRNA表达差异着手,采用以消减杂交为思路的多种分子生物学手段,先通过消减获得特异表达和缺失的基因片段,然后进行染色体定位乃至获得全基因。关于AIS病变部位不同组织的mRNA和蛋白质表达差异方面的研究已有许多报道,研究集中在对侧弯脊柱凸凹侧椎间盘内的Ⅰ、Ⅱ、X型胶原,TGF-?1、bFGF、成骨细胞蛋白质表达差异和ⅠⅡ、Ⅸ型胶原mRNA的表达差异方面,虽然均报道存在表达差异,但和组织学研究结果一样,这种差异“是因是果”难以确定,无法确定真正的病因。这种现状的原因是研究设计都是采用凸、凹侧对比,如果想找出真正的组织、蛋白或mRNA层面的差异,需要进行先证者自身发病前后对比研究,或先证者与正常人之间的对比研究,但是这在临床上是很难做到的,所以结果很难取得突破,那么只有寄希望于“反向”研究了。
“反向遗传学”策略既医学分子生物学策略是应用定位克隆遗传分析法对疾病基因进行染色体定位和疾病基因的筛查与鉴定。这种方法是基于任何一个基因必定在染色体上有位点,基因所决定的遗传学性状与这些位点有密切联系的遗传学原理,通过遗传学分析,获得某些疾病基因在染色体上的位置信息,然后根据这一位置信息,应用“物理图”的物理标记将遗传学信息转变为“物理标记”所代表的明确的染色体区域,再以相关区域的“相连片段群”来筛选可表达的基因,建立该区域的“转录图”。最后,通过“定位候选克隆”和“功能候选克隆”方法比较病人与正常人这些结构基因的区别以确定与疾病相关的改变,筛查与鉴定出疾病相关基因。关于AIS疾病基因的定位克隆研究共检索到7篇文献,由于各个研究应用了不同的研究群体和遗传统计学分析方法,各家报道的染色体定位的位点各异,分别在染色体6、9、10q、16、17、17p11、19p13.3、19 p 13和Xq21上,还没有取得公认的定位结果。
值得关注的是在定位克隆法研究中,有2位学者将疾病基因位点定在19p13 ,Alden研究的受试对象是美国人。Chan的报道不仅定位在19p13.3内,还进一步的精确定位于D19S216标志附近,在D19S894～D19S1034之间的5.2cM遗传距离,物理图约1.9Mb。研究的受试对象是中国广东人,采用400多个荧光标记的微卫星标志进行全基因组筛查,选用了参数和非参数两种相关分析的统计方法,基本排除了遗传模式对结果的影响,从一个家系中得到的结果在另外6个家系中得到验证,样本量大。文章的影响因子高达10.0以上,结果可信。此项研究虽然精确的定位到D19S216标志附近,但仍然属于疾病基因的染色体定位研究,未见后续的关于疾病基因的筛查与鉴定的研究报道。
我们认为承认Chan的研究结果,在其研究基础上做进一步的研究——候选基因筛查,以期在AIS的疾病基因定位研究上取得突破性进展是值得的,是有创新的和有科学价值的。为了确定19p13.3内基因的数目,检索在3亚带内有198个已知基因,其中只有17个是已知疾病的基因,尚无与AIS相关的基因报道。
如果要对3亚带内的198个基因进行筛查,从哪个基因开始是值得研究的。当然是选与AIS最相关的,Chan的精确区域定位是在D19S216标志附近,在物理图上查SH3GL1恰位于此区域内, Chan在报道中也指出SH3GL1是与破骨细胞代谢有关,可能是AIS重要的致病基因。
首选SH3GL1。关于基因SH3GL1功能的研究报道不多,Wise等人报道基因SH3GL1与细胞吞噬作用有关,参与细胞内吞、外排偶联的Ca2+通道的调节。也有研究认为SH3GL1与人类的白血病有关。
所以本项研究的目的一是对中国重庆AIS进行遗传流行病学调查分析,探讨遗传因素在发病中的作用和可能的遗传模式。二是采取定位候选克隆方法对AIS的疾病候选基因SH3GL1的外显子进行序列比对分析,研究是否存在碱基变异,为疾病基因筛查与鉴定做方法学准备。
研究方法:
1.遗传流行病学调查:设计AIS遗传流行病学调查表,收集在西南医院诊治过的先证者资料。确立诊断标准为站立前后位X线片Cobb角>10o为受累者, Cobb角<10o为易感者,年龄小于12岁的表型正常者为未确定者。由经过培训的调查员对先证者及其家系成员进行询问调查并填写调查表。方法为门诊、病房询问与电话询问相结合,为防止存在回忆偏倚对先证者、父亲和母亲分别进行询问记录。对怀疑受累者和易感者要进行临床望诊、Adam弯腰试验检查,必要时经过X线片检查确定。
2.遗传流行病学统计分析:对遗传流行病学调查数据采用SPSS10.0统计软件进行分析。通过年龄发病率分析,家族聚集性分析,遗传度计算(Falconer法)分析确定遗传因素的大小。根据单基因和多基因遗传病的特点分析确定遗传模式。
3.定位候选克隆:设计以家系为基础的“病例同胞对”研究方案,采集“同胞对”的血液标本,提取基因组DNA,根据基因SH3GL1核酸序列,以10个外显子为重点设计引物对,进行PCR扩增、克隆和测序。
4.序列比对分析:用GeneTool软件对外显子测序结果,进行“同胞对”之间和同胞对与NCBI之间的序列比对分析,判断是否存在碱基变异,并进行蛋白质结构预测分析。
研究结果:
1.遗传流行病学调查:设计了AIS遗传流行病学调查表,并成功的对73个家系按调查表进行遗传流行病学调查。
2.遗传流行病学统计:年龄发病率分析:发病75%在11～14岁之间,平均年龄10.84岁。家族聚集性分析:病例家族总发病率为4.86%,一级亲属的发病率为16.93%,二级亲属的发病率为7.21%,三级亲属发病率4.36%,而参照发病率为2.25%,统计学分析差异有显著意义(u=6.451,P﹤0.01 )。遗传度计算:先证者一级亲属的遗传度84%、二级亲属的遗传度88%和三级亲属的遗传度85%,经显著性检验(P<0.01)各级亲属遗传度有统计学意义。
3.定位候选克隆:候选基因SH3GL1中的10个外显子成功的在AIS“同胞对”的基因组DNA中得到扩增和克隆,并且序列测定均取得阳性结果。
4.序列比对分析:通过比对分析在候选基因SH3GL1的10个外显子中,共发现12处碱基变异,分别位于第2(2处)、4、5(3处)、6、8和10(2处,非编码区)六个外显子上。其中先证者基因编码mRNA第515位碱基如果为T,则形成终止密码子(TAG),导致蛋白阅读框发生改变(可能为107-518区域碱基,或为707-1213碱基),蛋白质序列预测分析显示编码截短蛋白,从而影响蛋白质的一级结构。
结论:
1.AIS是遗传病:年龄发病率分析显示75%在11～14岁之间,比较集中,属于遗传病的特点;病例家族发病率和先证者一级亲属、二级亲属和三级亲属的发病率均明显高于参照群体平均发病率,具有明显的家族聚集性;一级、二级和三级亲属的遗传度均大于84%,证明AIS与遗传显著相关。综合上述分析结果证明AIS是遗传病。
2.AIS是多基因遗传病:先证者的各级亲属发病率均明显高于参照人群发病率;同胞发病率远比1/2或1低;与先证者的亲缘关系越近,发病率越高,有明显的家族性聚集倾向。综合分析证明AIS属于多基因遗传病。
3.基因SH3GL1是AIS的致病基因之一:碱基序列比对分析证明外显子区域内的10处碱基变异位于编码区属于结构基因的变异,2处位于非编码区内的变异可能处于调控区域,先证者SH3GL1第5外显子的第515位碱基如果发生C→T突变,将形成终止密码子,导致蛋白阅读框发生改变,通过氨基酸序列预测分析,编码截短蛋白,从而显著影响蛋白质的结构。由此可以证明基因SH3GL1是AIS候选基因之一。
Background and Objects of the Study
Adolescent Idiopathic Scoliosis (AIS) is a three-dimensional deformation of dorsal spine structural perversion with vertebral body rotation. The incidence of children and teenagers is 2%~5% from reports abroad while incidence is 1.5%~3% from domestic reports. The incidence of severe deformation is 0.2~0.5% with female patients obviously more than male ones. Though ALS ususlly does not cause severe pathologic state except deformation, severe thoracocyrtosis may cause heart and lung hypofunction, leading to conspicuous somatotype deformation and even life. At present, the available treatments focus on correcting and curing on deformation including clinical observation, orthosis treatment, and operation on patients with lateral curvature above 45o. However, the treatment process was complicated with spinal deformity and dorsal spine activity obstacle left, producing no ideal effect. The ideal effect is to eliminate deformity. If diagnosis and cure on etiological factor can be made before symptoms and birth, or even gene diagnosis and cure can be made for pregnant women with family medical history before 6 days of ferrtilization and embryo nidation, spinal deformity will be cut off and AIS eliminated clinically. But etiological factor and invasive gene mechanism need to be identified first.
So far, The definite etiological factor of AIS has not been detected. Past researches concentrated on pathogenic mechanism of histology factors such as cartilage, ossature, nerves and muscle, but no definite conclusions were drawn. With unceasing deepening and progress of researches on medical genetics and medical molecular biology, clinically pathogenesy of many agnogenic common diseases has been proved relevant with heridity and virulence gene. At present, medical molecular biology holds that generation of various kinds of diseases is related with one or many genes. Thus people are easy to think that whether AIS is concerned with heredity? Recently many scholars deem AIS correlates with heredity. If it is related with heredity, what is the heredity mode? monogenic inheritance mode or polygenic inheritance mode? How is the genic location and molecule mechanism of disease? The approach to influence of genetic factors on disease and possible hereditary modes is the task of genetic epidemiology, while the approach to disease genic location and mechanism evoking disease, the research task of medical molecular biology.
There are few research reports about AIS epidemiology and genetic epidemiology, but these reports show familial aggregation. Genetic factor was deemed to be involved with AIS, which has several inheritance modes including autosomal dominant inheritance mode, X-linked dominant inheritance mode, and polygenic inheritance mode. That is to say, no final conclusion is drawn about whether AIS is genetic disorder and what kind of hereditary mode it is. No definit hereditary mode bring obstacle for chromosomal assignment study on AIS pathological gene The screening and identification of pathological gene were also affected consequently. Different analytical methods were applied in accordance with corresponding inheritance mode during the process of Chromosomal assignment analysis on pathological gene. Linkage analytical method was applied for monogenic disorder while correlation analysis for polygenic disease. The analytical results were indefinite with indefinite inheritance mode, which gave rise to different research results of Chromosomal assignment study. The decisive role was played by discussions on the influence of genetic factor on AIS and the possible inheritance mode in Chromosomal assignment study on AIS pathological gene as well as screening and identification of pathological gene. Only by determining AIS genetic factor and inheritance mode can Chromosomal assignment of pathological gene be conducted by employing linkage analysis (analytical method aiming at monogenic inheritance disorder) or correlation analysis (analytical method aiming at polygenic inheritance disorder) under the guidance of the inheritance mode.
Some studies on AIS molecular genetics mechanism have been reported. Two strategies (Norientation or uorientation ) were employed to the study. However the study is still at the stage of early chromosomal assignment study of the disease gene locus, and up to now a consensus gene locus has not been determined. Norientation strategy (classical genetics), namely the cloning methods based on gene function, was oriented to function of proteic substance or differential gene expression of mRNA. Multiple molecular biology methods oriented to differential hybridization were applied. Initially specific expression and the lacked gene fragment were acquired by subtraction, and then chromosomal assignment ending with obtaining whole gene was conducted. Studies on mRNA or differential expression of proteic substance of different tissues of AIS diseased region have been reported. They focused onⅠ、Ⅱ、X type collagen, TGF-?1、bFGF and osteoblast protein expression differences inside intervertebral cartilages in lateral curvature dorsal spine protruding and curved side andⅠ、Ⅱ、Ⅸtype collagen mRNA expression difference. Although it has been unanimously reported that differential expression existed, yet it has not been determined whether the differential expression was etiological factor or the result of disease. That is to say, the genuine etiological factor has not been detected. The reason of this status quo is that research and design use contrast of protruding and curved side. If differences of genuine tissue, protein or mRNA are needed to be found, contrast research of proband before and after invasion or research between proband and normal people should be made. But it is hard to make clinically, so the result is hard to break through with the only hope for“reversing”research.
Positional cloning genetic analysis was applied to the chromosomal assignment, screening, and identification of disease gene in the strategy of reverse genetics (molecular genetics). The method was premised on the genetics principles that any gene has its gene locus in chromatin body, and genetics characters determined by genes were closely correlated with these gene loci. position information of disease gene in chromatin body was obtained through genetics analysis, then according to the position information, the physical marking of“physical map”was applied to the transformation from genetics information to definite chromatin region represented by the physical marking, then the expressible gene was screened in accordance with linked gene fragment grouping of the correlated regions, and transcription map of the region could be made. Finally two methods of positioning candidate cloning and function candidate cloning were employed to compare the differences of architectural gene between the patients and the healthy one, thus changes correlated with the disease were defined, and disease gene was screened and identificated. 7 reports about positional cloning study on AIS disease gene has been retrived, yet a consensus conclusion has not been drawn due to different study groups, genetics statistical analytical method, and gene loci of chromosomal assignment. No received positioning results were got respectively for chromosome 6、9、10q、16、17、17p11、19p13.3、19 p 13 and Xq21.
It should be noticed that two scholars positioned the disease gene locus in 19p13 in their positional cloning studies. Their trail objects were Americans and Cantonese respectively. The key-gene locus of the latter was 19p13.3, and minor gene locus was 2p, which were approved to be located in the vicinity of the mark of D19S216 , ranging in the breadth of 5.2cM between D19S894～D19S1034 with the approximate length of 1.9Mb through further accurate positioning and recombination. Research objects that receive experiment are Cantonese. More than 400 fluorescently-labeled microsatellite signals were applied to make the whole genome screening, and two correlation analysis statistical methods of parameter and nonparameter were adopted. Influence of hereditary mode on results was basically eliminated. Besides, results from one family constellation were checked in other 6 family constellations with large sample size. Affectoi of article is as high as over 10.0, producing a convincing result. Although the disease gene locus was accurately positioned in the vicinity of the mark of D19S216 in the study, the research was still in the province of chromosomal assignment study of disease gene. The following screening and identification of pathological gene were not reported, which adequately approved that the gene locus was highly likely to correlate with AIS. Methods of positioning candidate cloning were worthy applying to the study on screening and identification of 198 genes in the position area.
To make sure the number of intragenes 19p13.3, 198 known genes inside 3 subband are retrived, among which only 17 are genes of known diseases. Gene reports related with AIS has not been attained.
If 198 genes inside 3 subband are to be screened, which gene to be start with were worthy studie. Certainly, gene that is the most related with AIS is chosen. Accurate positioning of Chan is in the vicinity of the mark of D19S216. From physical map, SH3GL1 is just located in this area. In report, Chan also pointed that SH3GL1 is concerned with osteoclast metabolism, and may be the important virulence gene of AIS.
SH3GL1 is the first choice.There are not many research reports about function of gene SH3GL1. People like Wise reported that gene SH3GL1 is revelant with cytophagocytesis, and participated in adjustment of Ca ion channel of cell internalization and excretion couplant. Some researches also think that SH3GL1 is related with leukemia in human beings.
Therefore the study was designed to make genetic epidemiologic survey and analysis of AIS in Chongqing and discuss the effect of inheritance factors on the AIS and the possible inheritance mode. Moreover, the method of positioning candidate cloning was applied to the analytical study on the Exon Sequence Alignment of the AIS Disease Candidate Gene SH3GL1, making technology preparation for the screening and identification of the disease gene.
Research Method
1.Genetic epidemiologic survey: The genetic epidemiology questionnaire for AIS was designed, and the data of index case who received medical treatment in Southwest Hospital were collected. Diagnostic code: affected individual: std. A-P position X-ray Cobb Ang > 10o; susc.: Cobb Ang <10o; the undetermined : normal phaenotype and age under 12. Initially trained investigators inquired index cases and their family members, and then filled the questionnaires. Investigating methods include out-patient clinic, hospital room inquiry, and telephone inquiry. Index cases and their fathers and mothers were inquired respectively to avoid memory errors. The suspected affected individual and susc. were received clinical inspections, Adam waist-bending test, and X-ray check when necessary.
2. Statistical analysis of genetic epidemiology: SPSS10.0 statistics software was applied to the analysis of the genetic epidemiology survey data. The influence of genetic factor was determined by analyses on disease age incidence, familial aggregation, and calculation of degree of heritability. Inheritance mode was determined according to analysis of features of the disease of monogenic or multifactorial inheritance.
3. Positioning candidate cloning:“case-sibling control design”research scheme based on family constellation was designed. A sib pair was choosed to collect blood preparation, and genome DNA was extracted. According to nucleotide sequence of gene SH3GL1, primer pair for PCR amplification, cloning, and sequencing with 10 exons as emphasis was designed.
4. Sequence comparative analysis: sequence comparative analysis for exon sequencing result between sib pairs, and that between sib pair and NCBI were conducted through GeneTool software to judge whether basic group mutation occurred or not. amino acid sequence comparative analysis for predict was made.
Research Results
1. Genetic epidemiologic survey: The genetic epidemiology questionnaire for AIS was designed and genetic epidemiology survey on 73 family constellations was conducted successfully according to the questionnaire.
2. Statistical analysis of genetic epidemiology: analysis of disease age incidence: morbility of the disease of 75% occurred between 11and 14. The mean morbility age was 10.84. Analysis of familial aggregation: family total disease incidence of the case was 4.86%. Disease incidence of first degree relative was 16.93%, and disease incidence of second degree relative was7.21%, and disease incidence of third degree relative was 4.36%, and disease incidence of the reference family was 2.25%. The incidences were significant differences by statistics analysis. (u=6.451,P﹤0.01 )Calculation of degree of heritability (Falconer Method): degree of heritability of first degree relative was 84%, degree of heritability of second degree relative was 88%, and degree of heritability of third degree relativ was above 85%, which had statistical significance through significance test (P<0.01)
3. Positioning candidate cloning: 10 exons of the candidate gene SH3GL1 were successfully amplified and cloned in genome DNA of an AIS sib pair, and the sequencings obtained positive results.
4. Sequence comparative analysis: 12 basic group mutations were found in 10 exons of the candidate gene SH3GL1 of patients with AIS. These mutations were located in the second exon (2 mutations), the fourth exon, the fifth exon (3 mutations), the sixth exon, the eighth exon, and the tenth exon (2 mutations, noncoding region). If basic group in 515 of mRNA was mutationed to T, stop codon(TAG) was made and open reading frame was altered , the sequence of proteinum was predicted and show brachytmema proteinum was encoded which could cause changes of primary structure.
Conclusions:
1. AIS was genetic disorder. Morbility of the disease usually occurred between 11 and 14 according to the analysis of disease age incidence. The family disease incidence of the case, disease incidence of first degree relative and the second degree relative and third degree relative were found obviously higher than the average disease incidence of the reference group. AIS was found to have obvious familial aggregation. The degree of heritability of first degree relative and second degree relative and third degree relativ was above 84%, which approved AIS was significantly correlated with heredity. AIS belonged to genetic disorder through comprehensive analysis. 2.AIS was a disease of multifactorial inheritance. On the whole, AIS was a genetic disorder; and the disease incidences of the index case’s all degrees of relatives were found to be higher than that of the reference crowd. Sibling disease incidence was found far lower than 1/2 or 1. The closer blood relationship with the index case means higher disease incidence. The disease has obvious familial aggregation. Among 73 family constellations, only 3 of them were found to have the family history of the disease. Index cases in other family constellations were all single cases. AIS belonged to the category of a disease of multifactorial inheritance through comprehensive analysis.
3. Gene SH3GL1 was one of the virulence genes of AIS. sequence comparative analysis show that Basic group mutations were found in exons of the candidate gene SH3GL1, and 10 of the mutations were located in the coding region and 2 of the mutations were located in the noncoding region. If basic group in 515 of mRNA was mutationed from C to T, stop codon(TAG) was made and open reading frame was altered , the sequence of proteinum predict show brachytmema proteinum was encoded which could cause changes of primary structure. It was demonstrated that Gene SH3GL1 was one of the virulence genes of AIS.
引文
[1] Harrlngton PR. the etiology of idiopathic scoliosis Clin Orthop,1977,l26:17
[2] Wynne R. A genetic survey of idiopathic scoliosis in Boston.Massachusetts. J Bone Joint Surg Am ,1973,55:974-982
[3] Kesling KL,Reinker KA. Scoliosis in twins:a meta-analysis of the literature and report of six cases.Spine,1997,22:2009
[4] Cart AJ. Adolescent idiopathic soliosis in identical twins. J Bone Joint Surg Br,1990,72:l077
[5] Gaertner RL. Idiopathic soliosis in identical(mono-zygotic)twins.South Med J,l979,72:231
[6]彭明惺,刘利君,周素华,等.脊柱侧弯的遗传流行病学调查.中华小儿外科杂志,1990,11(6):324-326
[7] Bonaiti C, Feingold J,Briard ML, et al. Genetics of idiopathic scoliosis [in French]. Helv Paediatr Acta, 1976,31:229–240
[8] Carr AJ. Adolescent idiopathic scoliosis in identical twins. J Bone Joint Surg,1990,72B:1077
[9] Czeizel A, Bellyei A, Barta O, et al. Genetics of adolescent idiopathic scoliosis. J Med Genet , 1978, 15:424–427
[10] Filho NA, Thompson MW. Genetic studies in scoliosis [abstract]. J Bone Joint Surg , 1971, 53 A:199
[11] Garland HG. Hereditary scoliosis. BMJ , 1934, 1:328
[12] Kesling KL, Reinker KA. Scoliosis in twins. A meta-analysis of the literature and report of six cases. Spine, 1997, 22:2009–14
[13] Wynne-Davies R. Familial (idiopathic) scoliosis. A family survey. J Bone Joint Surg, 1968, 50B:24–30
[14] Cowell HR, Hall JN, MacEwen GD. Familial patterns in idiopathic scoliosis [abstract]. J Bone Joint Surg , 1969, 51A:1236
[15] Jusctice CM,Miller NH,Marosy B,et a1.Familial idiopathic scoliosis:evidence of an X-linked susceptibility locos.Spine,2003, 28:589-594
[16] Falconer, D.S.The inheritance of liability to certain diseases, estimated from the incidence among relatives. Ann. Hum. Genetics. 1965,29(1): 51–76
[17]邱贵兴,庄乾宇.青少年特发性脊柱侧弯的流行病学研究进展.中华医学杂志,2006,86(11):790-792
[18]张奎星,朱鼎良,黄薇.多基因遗传病基因研究的策略和方法.生理科学进展,2001,32(3):215-219
[19]王以朋,叶启彬,吴波,等.北京地区脊柱侧弯患病率普查报告.中华流行病学杂志,1996,17(3):160-163
[20]孟令权,孟令凯,敖邦华,等.廊坊地区中、小学生脊柱侧弯患病率调查.医学理论与实践,2003,16(5):516-518
[21]于志悌,曲振海,王明胜,等.农村中小学生脊柱侧弯的普查及早期诊断与治疗.中华骨科杂志,1995,15(7):418-421
[22]马讯,赵斌,林庆宽,等.广西省对中小学生脊柱侧弯患病率调查.中华流行病学杂志,1995,16(2):109-110
[23]程斌,李锋涛,宋金辉.西安市25725名中小学生脊柱侧弯患病率调查.中国临床康复,2006,10(8):8-9
[24]张光铂,李子荣,魏新荣,等.学校青少年脊柱侧凸普查与治疗-北京市区、近郊区20418例普查报告.中华骨科杂志,1989,9(1):43-46
[25]刘尚礼,李卫平,李远景,等.广东省青少年脊柱侧凸患病率调查报告.中国脊柱脊髓杂志,2002,12(1):41-42
[1] Bonaiti C, Feingold J, Briard ML, et al. Genetics of idiopathic scoliosis [in French]. Helv Paediatr Acta, 1976,31:229-240
[2] Carr AJ. Adolescent idiopathic scoliosis in identical twins. J Bone Joint Surg, 1990,72B:1077
[3] Czeizel A, Bellyei A, Barta O, et al. Genetics of adolescent idiopathic scoliosis. J Med Genet ,1978,15:424-7
[4] Filho NA, Thompson MW. Genetic studies in scoliosis [abstract]. J Bone Joint Surg ,1971, 53 A:199
[5] Garland HG. Hereditary scoliosis. BMJ ,1934,1:328
[6] Kesling KL, Reinker KA. Scoliosis in twins. A meta-analysis of the literature and report of six cases. Spine, 1997,22:2009-2014
[7]何海龙,吴志宏,仉建国,等.X型胶原基因在特发性脊柱侧凸患者顶椎椎间盘内表达的初步研究.中华医学杂志,2004,84(20):1681-1685
[8]何海龙,吴志宏,仉建国,等.Ⅸ型胶原基因在特发性脊柱侧凸患者顶椎椎间盘内表达的初步研究.中华外科杂志,2005,43(8):513-516
[9]林琦吴志宏刘勇,等.特发性脊柱侧凸患者椎间盘中Ⅰ、Ⅱ型胶原基因mRNA的表达.中华外科杂志,2004,42(20):1268-1269
[10]林琦吴志宏刘勇,等.特发性脊柱侧凸患者椎间盘中X型胶原基因的表达.中国医学科学院学报,2004,26(6):696-699
[11]吴志宏翁习生郭树彬,等.青少年特发性脊柱侧弯椎间盘终板抑制消减cDNA文库的构建.中华外科杂志,2003,41(11):873
[12]徐宏光,邱贵兴,王以朋,等.青少年特发性脊柱侧凸椎体软骨终板核心蛋白、转化生长因子13-及碱性成纤维细胞生长因子的表达.中华外科杂志,2004,42(23):1460-1461
[13]徐宏光,邱贵兴,王以朋,等.青少年特发性脊柱侧凸椎体软骨终板核心蛋白和细胞因子的变化.中华骨科杂志,2005,25(12):725-730
[14]钱文伟,邱贵兴,吴志宏,等.青少年特发性脊柱侧凸患者顶椎关节突中I型胶原的表达.中国医学科学院学报,2005,72(2):138-142
[15]邱贵兴,郭盛杰,刘勇,等.脊柱侧凸关节突中Ⅰ、Ⅱ型胶原和转化生长因子TGF-?1表达的.中华医学杂志,2005,85(34):2391-2394
[16] Bertram H;,Steck E, Zimmermann G,et a1.Accelerated intervertebral disc d egeneration in scoliosis versus physiological ageing develops against a background of enhanced anabolic gene expression.Biochemical and biophysical research communications,2006,3:963-972
[17] Alden KJ, Marosy B, Nzegwu N, et al. Idiopathic scoliosis: identification of candidate regions on chromosome 19p13. Spine, 2006,31(16):1815-9
[18] Chan V, Fong GCY, Luk KDK, et al. A genetic locus for adolescent idiopathic scoliosis linked to chromosome 19p13.3.Am J Hum Genet. 2002, 71(2):401-6.
[19] Wise CA,Barnes R,Gillum J,et a1.Iocalization of susceptibility to familial idiopathic scoliosis.Spine,2000,25:2372-2380
[20] Salehi LB , Manquino M , DeSerio S , et a1 . Assignment of a locus for autosomaldominant idiopathic scoliosis(IS) to haman chromosome 17p11 . Hum Genet,2002,11l:40l-404
[21] Justice CM, Marosy B. Identification of candidate regions for familial idiopathic scoliosis. Spine.,2005, 10:1181-712
[22] Miller NH, Justice CM, Marosy B.Identification of candidate regions for familial idiopathic scoliosis. Spine, 2005,30(10):1181-712
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[4]孟令权,孟令凯,敖邦华,等.廊坊地区中、小学生脊柱侧弯患病率调查.医学理论与实践,2003,16(5):516-518
[5]于志悌,曲振海,王明胜,等.农村中小学生脊柱侧弯的普查及早期诊断与治疗.中华骨科杂志,1995,15(7):418-421
[6]马讯,赵斌,林庆宽,等.西省对中小学生脊柱侧弯患病率调查.中华流行病学杂志,1995,16(2):109-110
[7]程斌,李锋涛,宋金辉,等.西安市25 725名中小学生脊柱侧弯患病率调查.中国临床康复,2006,10(8):8-9
[8]张光铂,李子荣,魏新荣,等.学校青少年脊柱侧凸普查与治疗—北京市区、近郊区20418例普查报告.中华骨科杂志,1989,9(1):43-46
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[29]林琦吴志宏刘勇,等.特发性脊柱侧凸患者椎间盘中Ⅰ、Ⅱ型胶原基因mRNA的表达.中华外科杂志,2004,42(20):1268-1269
[30]林琦吴志宏刘勇,等.特发性脊柱侧凸患者椎间盘中X型胶原基因的表达.中国医学科学院学报,2004,26(6):696-699
[31]吴志宏翁习生郭树彬,等.青少年特发性脊柱侧弯椎间盘终板抑制消减cDNA文库的构建.中华外科杂志,2003,41(11):873
[32]徐宏光,邱贵兴,王以朋,等.青少年特发性脊柱侧凸椎体软骨终板核心蛋白、转化生长因子13-及碱性成纤维细胞生长因子的表达.中华外科杂志,2004,42(23):1460-1461
[33]徐宏光,邱贵兴,王以朋,等.青少年特发性脊柱侧凸椎体软骨终板核心蛋白和细胞因子的变化.中华骨科杂志,2005,25(12):725-730
[34]钱文伟,邱贵兴,吴志宏,等.青少年特发性脊柱侧凸患者顶椎关节突中I型胶原的表达.中国医学科学院学报,2005,72(2):138-142
[35]邱贵兴,郭盛杰,刘勇,等.脊柱侧凸关节突中Ⅰ、Ⅱ型胶原和转化生长因子TGF-?1表达的.中华医学杂志,2005,85(34):2391-2394
[36] Bertram H;,Steck E, Zimmermann G,et a1.Accelerated intervertebral disc d egeneration in scoliosis versus physiological ageing develops against a background of enhanced anabolic gene expression.Biochemical and biophysical research communications,2006,3:963-972
[1] Qiu Guixing, Zhuang Qianyu. Progress of genetic epidemiology investigation on Adolescent Idiopathic Scoliosis. Chinese Medical Journal, 2006, 11:790-792
[2] Falconer DS.The inheritance of liability to certain diseases,estimated from the incidence among relatives.Ann. Hum. Genet.1965,29: 51–76
[3] Harrlngton PR. the etiology of idiopathic scoliosis in Orthop,1977,l26:17
[4] Wynne R. A genetic survey of idiopathic scoliosis in Boston.Massachusetts. J Bone Joint Surg Am ,1973,55:974-982
[5] Kesling KL,Reinker KA. Scoliosis in twins:a meta-analysis of the literature and report of six cases.Spine,1997,22:2009-2014
[6] Cart AJ. Adolescent idiopathic soliosis in identical twins. J Bone Joint Surg Br,1990,72:l077
[7] Gaertner RL. Idiopathic soliosis in identical(mono-zygotic)twins.South Med J,l979,72:231
[8] Pen Mingxing, Liu Lijun, Zhou suhua, etc. genetic epidemiology investigation of scoliolosis. Chinese pediatric surgery journal, 1990, 6:324-326
[9] Bonaiti C, Feingold J, Briard ML, et al. Genetics of idiopathic scoliosis [in French]. Helv Paediatr Acta, 1976,31:229–240
[10] Carr AJ. Adolescent idiopathic scoliosis in identical twins. J Bone Joint Surg, 1990,72B:1077
[11] Czeizel A, Bellyei A, Barta O, et al. Genetics of adolescent idiopathic scoliosis. J Med Genet ,1978,15:424–427
[12] Filho NA, Thompson MW. Genetic studies in scoliosis [abstract]. J Bone Joint Surg ,1971, 53 A:199
[13] Garland HG. Hereditary scoliosis. BMJ ,1934,1:328
[14] Kesling KL, Reinker KA. Scoliosis in twins. A meta-analysis of the literature and report of six cases. Spine, 1997,22:2009–2014
[15] Wynne-Davies R. Familial (idiopathic) scoliosis. A family survey. J Bone Joint Surg ,1968,50B:24–30
[16] Cowell HR, Hall JN, MacEwen GD. Familial patterns in idiopathic scoliosis [abstract]. J Bone Joint Surg ,1969,51 A:1236
[17] Jusctice CM,Miller NH,Marosy B,et a1.Familial idiopathic scoliosis:evidence of an X-linked susceptibility locos.Spine,2003,28:589-594
[18] Zhang Kuixing, Zhu Dingliang, Huang Wei. Strategy and method of gene study of disease of multifactorial inheritance. Physiological science progress, 2001, 3:215-219
[2] Wynne R. A genetic survey of idiopathic scoliosis in Boston.Massachusetts. J Bone Joint Surg Am ,1973,55:974-982
[3] Kesling KL,Reinker KA. Scoliosis in twins:a meta-analysis of the literature and report of six cases.Spine,1997,22:2009
[4] Cart AJ. Adolescent idiopathic soliosis in identical twins. J Bone Joint Surg Br,1990,72:l077
[5] Gaertner RL. Idiopathic soliosis in identical(mono-zygotic)twins.South Med J,l979,72:231
[6]彭明惺,刘利君,周素华,等.脊柱侧弯的遗传流行病学调查.中华小儿外科杂志,1990,11(6):324-326
[7] Bonaiti C, Feingold J,Briard ML, et al. Genetics of idiopathic scoliosis [in French]. Helv Paediatr Acta, 1976,31:229–240
[8] Carr AJ. Adolescent idiopathic scoliosis in identical twins. J Bone Joint Surg,1990,72B:1077
[9] Czeizel A, Bellyei A, Barta O, et al. Genetics of adolescent idiopathic scoliosis. J Med Genet , 1978, 15:424–427
[10] Filho NA, Thompson MW. Genetic studies in scoliosis [abstract]. J Bone Joint Surg , 1971, 53 A:199
[11] Garland HG. Hereditary scoliosis. BMJ , 1934, 1:328
[12] Kesling KL, Reinker KA. Scoliosis in twins. A meta-analysis of the literature and report of six cases. Spine, 1997, 22:2009–14
[13] Wynne-Davies R. Familial (idiopathic) scoliosis. A family survey. J Bone Joint Surg, 1968, 50B:24–30
[14] Cowell HR, Hall JN, MacEwen GD. Familial patterns in idiopathic scoliosis [abstract]. J Bone Joint Surg , 1969, 51A:1236
[15] Jusctice CM,Miller NH,Marosy B,et a1.Familial idiopathic scoliosis:evidence of an X-linked susceptibility locos.Spine,2003, 28:589-594
[16] Falconer, D.S.The inheritance of liability to certain diseases, estimated from the incidence among relatives. Ann. Hum. Genetics. 1965,29(1): 51–76
[17]邱贵兴,庄乾宇.青少年特发性脊柱侧弯的流行病学研究进展.中华医学杂志,2006,86(11):790-792
[18]张奎星,朱鼎良,黄薇.多基因遗传病基因研究的策略和方法.生理科学进展,2001,32(3):215-219
[19]王以朋,叶启彬,吴波,等.北京地区脊柱侧弯患病率普查报告.中华流行病学杂志,1996,17(3):160-163
[20]孟令权,孟令凯,敖邦华,等.廊坊地区中、小学生脊柱侧弯患病率调查.医学理论与实践,2003,16(5):516-518
[21]于志悌,曲振海,王明胜,等.农村中小学生脊柱侧弯的普查及早期诊断与治疗.中华骨科杂志,1995,15(7):418-421
[22]马讯,赵斌,林庆宽,等.广西省对中小学生脊柱侧弯患病率调查.中华流行病学杂志,1995,16(2):109-110
[23]程斌,李锋涛,宋金辉.西安市25725名中小学生脊柱侧弯患病率调查.中国临床康复,2006,10(8):8-9
[24]张光铂,李子荣,魏新荣,等.学校青少年脊柱侧凸普查与治疗-北京市区、近郊区20418例普查报告.中华骨科杂志,1989,9(1):43-46
[25]刘尚礼,李卫平,李远景,等.广东省青少年脊柱侧凸患病率调查报告.中国脊柱脊髓杂志,2002,12(1):41-42
[1] Bonaiti C, Feingold J, Briard ML, et al. Genetics of idiopathic scoliosis [in French]. Helv Paediatr Acta, 1976,31:229-240
[2] Carr AJ. Adolescent idiopathic scoliosis in identical twins. J Bone Joint Surg, 1990,72B:1077
[3] Czeizel A, Bellyei A, Barta O, et al. Genetics of adolescent idiopathic scoliosis. J Med Genet ,1978,15:424-7
[4] Filho NA, Thompson MW. Genetic studies in scoliosis [abstract]. J Bone Joint Surg ,1971, 53 A:199
[5] Garland HG. Hereditary scoliosis. BMJ ,1934,1:328
[6] Kesling KL, Reinker KA. Scoliosis in twins. A meta-analysis of the literature and report of six cases. Spine, 1997,22:2009-2014
[7]何海龙,吴志宏,仉建国,等.X型胶原基因在特发性脊柱侧凸患者顶椎椎间盘内表达的初步研究.中华医学杂志,2004,84(20):1681-1685
[8]何海龙,吴志宏,仉建国,等.Ⅸ型胶原基因在特发性脊柱侧凸患者顶椎椎间盘内表达的初步研究.中华外科杂志,2005,43(8):513-516
[9]林琦吴志宏刘勇,等.特发性脊柱侧凸患者椎间盘中Ⅰ、Ⅱ型胶原基因mRNA的表达.中华外科杂志,2004,42(20):1268-1269
[10]林琦吴志宏刘勇,等.特发性脊柱侧凸患者椎间盘中X型胶原基因的表达.中国医学科学院学报,2004,26(6):696-699
[11]吴志宏翁习生郭树彬,等.青少年特发性脊柱侧弯椎间盘终板抑制消减cDNA文库的构建.中华外科杂志,2003,41(11):873
[12]徐宏光,邱贵兴,王以朋,等.青少年特发性脊柱侧凸椎体软骨终板核心蛋白、转化生长因子13-及碱性成纤维细胞生长因子的表达.中华外科杂志,2004,42(23):1460-1461
[13]徐宏光,邱贵兴,王以朋,等.青少年特发性脊柱侧凸椎体软骨终板核心蛋白和细胞因子的变化.中华骨科杂志,2005,25(12):725-730
[14]钱文伟,邱贵兴,吴志宏,等.青少年特发性脊柱侧凸患者顶椎关节突中I型胶原的表达.中国医学科学院学报,2005,72(2):138-142
[15]邱贵兴,郭盛杰,刘勇,等.脊柱侧凸关节突中Ⅰ、Ⅱ型胶原和转化生长因子TGF-?1表达的.中华医学杂志,2005,85(34):2391-2394
[16] Bertram H;,Steck E, Zimmermann G,et a1.Accelerated intervertebral disc d egeneration in scoliosis versus physiological ageing develops against a background of enhanced anabolic gene expression.Biochemical and biophysical research communications,2006,3:963-972
[17] Alden KJ, Marosy B, Nzegwu N, et al. Idiopathic scoliosis: identification of candidate regions on chromosome 19p13. Spine, 2006,31(16):1815-9
[18] Chan V, Fong GCY, Luk KDK, et al. A genetic locus for adolescent idiopathic scoliosis linked to chromosome 19p13.3.Am J Hum Genet. 2002, 71(2):401-6.
[19] Wise CA,Barnes R,Gillum J,et a1.Iocalization of susceptibility to familial idiopathic scoliosis.Spine,2000,25:2372-2380
[20] Salehi LB , Manquino M , DeSerio S , et a1 . Assignment of a locus for autosomaldominant idiopathic scoliosis(IS) to haman chromosome 17p11 . Hum Genet,2002,11l:40l-404
[21] Justice CM, Marosy B. Identification of candidate regions for familial idiopathic scoliosis. Spine.,2005, 10:1181-712
[22] Miller NH, Justice CM, Marosy B.Identification of candidate regions for familial idiopathic scoliosis. Spine, 2005,30(10):1181-712
[1] Bunnell WP. The natural history of idiopathic scoliosis before skeletal maturity. Spine 1986;11:773–6
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[3]王以朋,叶启彬,吴波,等.北京地区脊柱侧弯患病率普查报告.中华流行病学杂志,1996,17(3):160-163
[4]孟令权,孟令凯,敖邦华,等.廊坊地区中、小学生脊柱侧弯患病率调查.医学理论与实践,2003,16(5):516-518
[5]于志悌,曲振海,王明胜,等.农村中小学生脊柱侧弯的普查及早期诊断与治疗.中华骨科杂志,1995,15(7):418-421
[6]马讯,赵斌,林庆宽,等.西省对中小学生脊柱侧弯患病率调查.中华流行病学杂志,1995,16(2):109-110
[7]程斌,李锋涛,宋金辉,等.西安市25 725名中小学生脊柱侧弯患病率调查.中国临床康复,2006,10(8):8-9
[8]张光铂,李子荣,魏新荣,等.学校青少年脊柱侧凸普查与治疗—北京市区、近郊区20418例普查报告.中华骨科杂志,1989,9(1):43-46
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[12] Bonaiti C, Feingold J, Briard ML, et al. Genetics of idiopathic scoliosis [in French]. Helv Paediatr Acta 1976;31:229-40
[13] Carr AJ. Adolescent idiopathic scoliosis in identical twins. J Bone Joint Surg 1990;72B:1077
[14] Czeizel A, Bellyei A, Barta O, et al. Genetics of adolescent idiopathic scoliosis. J Med Genet 1978;15:424-7
[15] Filho NA, Thompson MW. Genetic studies in scoliosis [abstract]. J Bone Joint Surg 1971;53A:199
[16] Garland HG. Hereditary scoliosis. BMJ 1934,1:328
[17] Kesling KL, Reinker KA. Scoliosis in twins. A meta-analysis of the literature and report of six cases. Spine 1997,22:2009-2014
[18] Riseborough EJ, Wynne-Davies R. A genetic survey of idiopathic scoliosis in Boston, MA. J Bone Joint Surg 1973;55A:974-982
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[20] Cowell HR, Hall JN, MacEwen GD. Familial patterns in idiopathic scoliosis [abstract]. J Bone Joint Surg 1969;51A:1236
[21] [21] Jusctice CM,Miller NH,Marosy B,et a1.Familial idiopathic scoliosis:evidence of an X-linked susceptibility locos.Spine,2003.28:589-594
[22]张奎星,朱鼎良,黄薇.多基因遗传病基因研究的策略和方法.生理科学进展.2001,32(3):215-219
[23] Harrlngton PR. the etiology of idiopathic scoliosis in Orthop,1977,l26:17
[24] Wynne R. A genetic survey of idiopathic scoliosis in Boston.Massachusetts. J Bone Joint Surg Am ,1973,55:974-982
[25] Cart AJ. Adolescent idiopathic soliosis in identical twins. J Bone Joint Surg Br,1990,72:l077
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[29] Falconer DS.The inheritance of liability to certain diseases,estimated from the incidence among relatives.Ann. Hum. Genet.1965,29: 51–76
[1] Bonaiti C, Feingold J, Briard ML, et al. Genetics of idiopathic scoliosis [in French]. Helv Paediatr Acta, 1976,31:229–240
[2] Carr AJ. Adolescent idiopathic scoliosis in identical twins. J Bone Joint Surg, 1990,72B:1077
[3] Czeizel A, Bellyei A, Barta O, et al. Genetics of adolescent idiopathic scoliosis. J Med Genet ,1978,15:424–427
[4] Filho NA, Thompson MW. Genetic studies in scoliosis [abstract]. J Bone Joint Surg ,1971,53A:199
[5] Garland HG. Hereditary scoliosis. BMJ ,1934,1:328
[6] Kesling KL, Reinker KA. Scoliosis in twins. A meta-analysis of the literature and report of six cases. Spine, 1997,22:2009–2014
[7] Wynne-Davies R. Familial (idiopathic) scoliosis. A family survey. J Bone Joint Surg ,1968,50B:24–30
[8] Cowell HR, Hall JN, MacEwen GD. Familial patterns in idiopathic scoliosis [abstract]. J Bone Joint Surg ,1969,51A:1236
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[15] Chan V,Fong GCY,Luk KDK.et a1.A genetic locus for adolescent idiopatlfic scoliosis linked to chromosome 19p13.3. Am J Hum Genet,2002,71:401-406
[16] Salehi LB,Manquino M,DeSerio S,et a1.Assignment of a locus for autosomaldominant idiopathic scoliosis(IS) to haman chromosome 17p11.Hum Genet,2002,11l:40l-404
[17] Justice CM, Marosy B. Identification of candidate regions for familial idiopathic scoliosis. Spine. 2005, 10:1181-712
[18] Miller NH, Justice CM, Marosy B. Identification of candidate regions for familial idiopathic scoliosis. Spine, 2005, 10:1181-712
[19] 19 Alden KJ, Marosy B, Nzegwu N, et al . Idiopathic scoliosis: identification of candidate regions on chromosome 19p13. Spine, 2006,16:1815-1819
[20] Bashiardes S, Veile R, Allen M, et al。SNTG1, the gene encoding gamma1-syntrophin: a candidate gene for idiopathic scoliosis. Hum Genet,2004 ,1:81-89
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[27]何海龙,吴志宏,仉建国,等.X型胶原基因在特发性脊柱侧凸患者顶椎椎间盘内表达的初步研究.中华医学杂志,2004,84(20):1681-1685
[28]何海龙,吴志宏,仉建国,等.Ⅸ型胶原基因在特发性脊柱侧凸患者顶椎椎间盘内表达的初步研究.中华外科杂志,2005,43(8):513-516
[29]林琦吴志宏刘勇,等.特发性脊柱侧凸患者椎间盘中Ⅰ、Ⅱ型胶原基因mRNA的表达.中华外科杂志,2004,42(20):1268-1269
[30]林琦吴志宏刘勇,等.特发性脊柱侧凸患者椎间盘中X型胶原基因的表达.中国医学科学院学报,2004,26(6):696-699
[31]吴志宏翁习生郭树彬,等.青少年特发性脊柱侧弯椎间盘终板抑制消减cDNA文库的构建.中华外科杂志,2003,41(11):873
[32]徐宏光,邱贵兴,王以朋,等.青少年特发性脊柱侧凸椎体软骨终板核心蛋白、转化生长因子13-及碱性成纤维细胞生长因子的表达.中华外科杂志,2004,42(23):1460-1461
[33]徐宏光,邱贵兴,王以朋,等.青少年特发性脊柱侧凸椎体软骨终板核心蛋白和细胞因子的变化.中华骨科杂志,2005,25(12):725-730
[34]钱文伟,邱贵兴,吴志宏,等.青少年特发性脊柱侧凸患者顶椎关节突中I型胶原的表达.中国医学科学院学报,2005,72(2):138-142
[35]邱贵兴,郭盛杰,刘勇,等.脊柱侧凸关节突中Ⅰ、Ⅱ型胶原和转化生长因子TGF-?1表达的.中华医学杂志,2005,85(34):2391-2394
[36] Bertram H;,Steck E, Zimmermann G,et a1.Accelerated intervertebral disc d egeneration in scoliosis versus physiological ageing develops against a background of enhanced anabolic gene expression.Biochemical and biophysical research communications,2006,3:963-972
[1] Qiu Guixing, Zhuang Qianyu. Progress of genetic epidemiology investigation on Adolescent Idiopathic Scoliosis. Chinese Medical Journal, 2006, 11:790-792
[2] Falconer DS.The inheritance of liability to certain diseases,estimated from the incidence among relatives.Ann. Hum. Genet.1965,29: 51–76
[3] Harrlngton PR. the etiology of idiopathic scoliosis in Orthop,1977,l26:17
[4] Wynne R. A genetic survey of idiopathic scoliosis in Boston.Massachusetts. J Bone Joint Surg Am ,1973,55:974-982
[5] Kesling KL,Reinker KA. Scoliosis in twins:a meta-analysis of the literature and report of six cases.Spine,1997,22:2009-2014
[6] Cart AJ. Adolescent idiopathic soliosis in identical twins. J Bone Joint Surg Br,1990,72:l077
[7] Gaertner RL. Idiopathic soliosis in identical(mono-zygotic)twins.South Med J,l979,72:231
[8] Pen Mingxing, Liu Lijun, Zhou suhua, etc. genetic epidemiology investigation of scoliolosis. Chinese pediatric surgery journal, 1990, 6:324-326
[9] Bonaiti C, Feingold J, Briard ML, et al. Genetics of idiopathic scoliosis [in French]. Helv Paediatr Acta, 1976,31:229–240
[10] Carr AJ. Adolescent idiopathic scoliosis in identical twins. J Bone Joint Surg, 1990,72B:1077
[11] Czeizel A, Bellyei A, Barta O, et al. Genetics of adolescent idiopathic scoliosis. J Med Genet ,1978,15:424–427
[12] Filho NA, Thompson MW. Genetic studies in scoliosis [abstract]. J Bone Joint Surg ,1971, 53 A:199
[13] Garland HG. Hereditary scoliosis. BMJ ,1934,1:328
[14] Kesling KL, Reinker KA. Scoliosis in twins. A meta-analysis of the literature and report of six cases. Spine, 1997,22:2009–2014
[15] Wynne-Davies R. Familial (idiopathic) scoliosis. A family survey. J Bone Joint Surg ,1968,50B:24–30
[16] Cowell HR, Hall JN, MacEwen GD. Familial patterns in idiopathic scoliosis [abstract]. J Bone Joint Surg ,1969,51 A:1236
[17] Jusctice CM,Miller NH,Marosy B,et a1.Familial idiopathic scoliosis:evidence of an X-linked susceptibility locos.Spine,2003,28:589-594
[18] Zhang Kuixing, Zhu Dingliang, Huang Wei. Strategy and method of gene study of disease of multifactorial inheritance. Physiological science progress, 2001, 3:215-219