Van Der Woude综合征的遗传学分析

目的 对4个Van der Woude综合征（VWS）家系和1个Popliteal pterygium综合征（PPS）家系进行进行遗传学分析。方法 对以上家系进行临床表型观察，计算表型分布频率和表型基因频率。结果 VWS表型分布频率为：腭裂或隐腭裂（82%），唇瘘（52%），缺牙（22%）；表型基因频率约为1/50000。结论 Van der Woude综合征表型变化可能是由于存在特定的修饰基因，进一步寻找修饰基因将为临床基因治疗和基因诊断提供分子基础。     

家族性Dravet综合征家系基因突变分析

目的探讨家族性Dravet综合征家系基因突变情况。方法 90例Dravet综合征患儿及其家属,采用PCR法提取外周血DNA,采用基因测序法和多重连接依赖的探针扩增技术对SCN1A基因突变进行筛查,并采用捕获测序癫痫基因检测包对无SCN1A基因突变的家系进行筛查。结果 90例患儿中62例发生SCN1A基因突变,SCN1A基因突变率68.89%,其中错义突变29例,截断突变26例,剪切位点突变5例,SCN1A基因片段重复或缺失2例;有5个家系携带SCN1A基因突变,其中3个家系属SCN1A遗传性杂合突变,来自母源,母亲临床表型是Dravet综合征或热性惊厥,余2个家系测序结果可能为父母一方为SCN1A突变嵌合体,父母临床表型正常。结论家族性Dravet综合征多是SCN1A基因突变导致。     

遗传性出血性毛细血管扩张症一家系致病基因分析及沙利度胺治疗疗效观察

目的:分析遗传性出血性毛细血管扩张症(hereditary hemorrhagic telangiectasia,HHT)一家系ACVRL1、ENG和MADH4(SMAD4)基因突变,并观察沙利度胺对其治疗的疗效。方法:对该家系进行详细的家系调查,采用PCR方法对先证者进行ACVRL1、ENG和SMAD4基因全部外显子序列扩增,应用Sanger测序法进行序列分析,将结果与数据库参照序列进行对比;对检测到的可疑突变在纳入研究的其他7名家系成员中进一步进行检测。先证者采用沙利度胺(100 mg/d)治疗6个月,而后观察其出血频率、出血量、输血频率等,评估疗效。结果:先证者(Ⅱ-1)及其另外4名家系成员(Ⅱ-2、Ⅲ-1、Ⅲ-2、Ⅲ-3)均存在有ACVRL1基因c.1231CT杂合突变,与临床表型共分离,为文献已经报道的突变;同时该家系部分成员(Ⅱ-1、Ⅱ-3,Ⅲ-1、Ⅲ-2)存在有ENG基因c.1096GC突变,不与临床表型共分离,为数据库收录的基因多态性;SMAD4基因未见突变。经沙利度胺治疗后,先证者出血频率、出血量、输血频率均减少,血红蛋白浓度和血清铁均升高。结论:HHT具有表型异质性,且随着年龄增长表型增多;ACVRL1基因c.1231CT突变是该家系的致病基因。沙利度胺对遗传性出血性毛细血管扩张症出血具有较好的疗效。     

汉族、维吾尔族献血人群CD61基因外显子10序列分析

目的了解CD61基因外显子10的突变及其在中国汉族、维吾尔族(简称维族)人群中的分布情况。方法对随机采集的149(人)份汉族和96(人)份维族献血者的血液标本,在扩增CD61基因外显子10后直接测序,分析外显子10编码区基因序列。结果在汉族和维族人群中均发现在CD61基因外显子10的1533、1545位分别存在较高频率的AG、GA突变。在汉族人群中还存在1529位CT。3个突变点均为同义突变,且1533和1545位的突变位于同一条染色体上。汉族人中1533A1545G表型频率为48.32%(72/149),1533G1545A表型频率为12.75%(19/149),1533A+G/1545G+A表型频率为38.93%(58/149),C1529T突变约占0.67%(1/149),维族中1533A1545G表型频率为53.12%(51/96),1533G1545A表型频率为5.22%(5/96),1533A+G/1545G+A表型频率为41.66%(40/96),无C1529T。结论中国汉族和维族人群的CD61基因外显子10存在3个SNP,且在汉族和维族人群分布频率差异甚小,这显示2个民族之间血缘上的关系比较亲近。     

洛阳地区汉族人群Duffy血型基因型分布

目的调查洛阳地区汉族人群Duffy血型的分布情况。方法收集洛阳本地汉族献血者样本571份,应用血清学方法进行Duffy血型表型鉴定,使用PCR-SSP技术进行基因分型。结果 571份样本中,Duffy血型各种表型分布为:Fy(a+b-)表型523例(91.59%),Fy(a+b+)表型42例(7.36%),Fy(a-b+)表型6例(1.05%),未发现Fy(a-b-)表型。基因频率为:Fya(0.952 7),Fyb(0.047 3)。结论洛阳地区汉族人群Duffy血型分布情况与国内其他地区汉族人群相似,与白种人和黑种人有明显的差别。     

青岛骨髓库HLA基因多态性研究

目的研究青岛骨髓库汉族人群HLAA、B、DRB1基因多态性分布。方法采用序列特异性引物PCR方法(SSPPCR)对1383名自愿骨髓捐献汉族进行HLAA、B、DRB1位点分型检测(低分辨率)。结果共检出HLA表型特异性A位点16个;B位点39个;DRB1位点14个,分别占各位点可检出表型特异性数的76%、91%和100%。用表型估算法对各位点的等位基因频率和双位点单型频率进行计算并进行有效的统计学处理。结论获得青岛骨髓库汉族人群HLAA、B、DRB1位点基因多态性资料。该库HLA多态性分布更接近中国北方汉族人群分布规律。     

北京地区献血人群Duffy血型表型筛查及稀有血型库的建立

目的调查Duffy血型表型在北京地区的分布频率,将筛选出的稀有血型红细胞甘油化冷冻保存,并建立稀有血型献血者档案资料,解决Duffy稀有血型患者输血难题。方法收集北京地区无血源关系健康汉族献血者的血液标本1 752份,少数民族献血者标本274份,外籍献血者标本137份,利用微柱凝胶法鉴定Duffy血型表型,PCRSSP检测Duffy血型基因,并随机选择17例做Duffy血型基因编码区域序列测定,以检测表现型与基因型符合性。结果1 752名汉族献血者,274名少数民族献血者,137名外籍献血者的Duffy血型各表型分布分别为:Fy(a+b-)表型1 530例(87.33%)、227例(82.85%)、83例(60.58%),Fy(a+b+)表型213例(12.16%)、42例(15.33%)、32例(23.36%),Fy(a-b+)表型9例(0.51%)、5例(1.82%)、20例(14.60%),Fy(a-b-)表型0、0、2例(1.46%)。FYA基因频率分别为93.41、90.51、72.26,FYB基因频率分别为6.59、9.49、26.28,FY基因频率0、0、1.46。汉族与少数民族献血者,中国与外籍献血者表型频率比较,差异均具有统计学意义(P0.05)。17例FY基因编码序列部分测序结果除2例Fy(a-b-)由于存在FYB基因外,其余标本与血清学、PCR-SSP法均符合。FYA与FYB基因的不同在于FY基因编码区域第131位核苷酸碱基为A或G。结论北京地区汉族献血者Fya阴性表型频率与国内报道相似,而少数民族及外籍献血者较高,在北京地区建立Fya阴性稀有血型供者库,以便解决稀有血型患者紧急输血问题,对提高输血安全具有重要意义。     

血型血清学ABO亚型95例的分子机制研究

目的研究95例中国人群血型血清学ABO亚型个体的分子机制。方法 2013年11月—2017年10月采用血型血清学方法对ABO正反定型不符标本进行检测;采用序列特异性引物聚合酶链反应(PCR-SSP)及第6、7外显子基因测序方法进行基因分型,其中部分标本采用了第1—7外显子的测序。结果对血型血清学方法确认的95例(73例先证者,22例家系成员)ABO亚型标本。经PCR-SSP和基因测序检测,73例ABO亚型先证者中检出A亚型基因的个体13例(17.8%),检出B亚型基因的个体17例(23.3%),检出B(A)型基因的个体22例(30.1%),共52例(71.2%,52/73);22例家系成员中检出ABO亚型基因者18例。先证者中未检出ABO亚型基因者21例(28.8%,21/73),且21例中5例(6.8%,5/73)检出O基因但表达弱A或弱B抗原;22例家系成员中4例未检出ABO亚型基因,共25例(25/95)。结论 ABO亚型的基因型与表型不完全符合,且同1个血型血清学表型对应不同的基因型,同1个基因型又表现为不同的血型血清学结果。     

B(A)血型表型与基因型研究

目的了解河北地区中国人群中B(A)血型的分布特点及其表型与基因型的关联性。方法采用血型血清学方法对ABO正反定型不符标本进行检测;采用序列特异性引物聚合酶链反应(SSP-PCR)及第6、7外显子基因测序方法进行基因分型,并对表型和基因型的关联性进行分析,并做家系调查。结果经测序确定为B(A)血型的有34例(22例B(A)血型个体,12例家系成员),包括两种B(A)等位基因,分别为B(A)02和B(A)04。22例B(A)血型个体中B(A)02有17(77.3%)例,B(A)04有5(22.7%)例。这两种等位基因所对应的表型包括B(A),AxB,A_2B和ABx。结论在河北地区中国人群中,此次主要检测出B(A)血型的两种基因型B(A)02和B(A)04,以B(A)02常见,基因型所对应的表型具有多样性。家系调查证实了B(A)血型的顺式遗传特性。     

遗传性凝血因子V缺陷症的诊断

目的对遗传性凝血因子V(FV)缺陷症进行诊断。方法通过检测先证者及家系成员的活化部分凝血活酶时间(APTT)、凝血酶原时间(PT)、FV促凝活性(FV∶C)和FV抗原(FV∶Ag)等进行临床表型诊断,应用聚合酶链反应(PCR)方法对先证者F5基因的25个外显子及其侧翼序列进行扩增,PCR产物进行直接测序以发现其基因突变,进行家系调查以期发现遗传规律。结果先证者APTT和PT显著延长,FV∶C为3.5%,FV∶Ag为1.47%。F5基因测序发现,先证者F5基因外显子区共有7处与GenBank AY364535序列不同的位点,其中外显子10区的C38591T杂合突变导致产生1个终止密码(R506Term),外显子13区的C47504T纯合变异导致编码氨基酸L1369F替换,该变异被证实为基因多态性。家系分析表明,前者遗传于先证者母亲,后者遗传于其父母双亲。结论通过表型检测、家系调查和基因诊断,明确该先症者为遗传性凝血因子V缺陷症,C38591T杂合突变产生终止密码是导致先证者FV缺陷的原因之一。     

胎儿硬腭的三维超声显像方法及其应用

目的探讨胎儿硬腭的三维超声显像方法及其在产前诊断胎儿腭裂中的应用价值。方法采集100胎孕13～35周正常胎儿及32胎唇裂胎儿颜面部的三维容积数据,通过旋转x、y、z轴、调节灰度阈值和片层,显示观察胎儿硬腭矢状面、冠状面和横断面声像图。结果正常胎儿硬腭矢状面显示率为93.00%(93/100),呈前后走向的带状稍强回声,表面光滑,前方与上牙槽突强回声相延续;冠状面显示率为91.00%(91/100),冠状面上硬腭位于中部,呈连续的弧形带状稍强回声,与上方鼻骨强回声构成三角关系;横断面显示率为90.00%(90/100),横断面上硬腭呈拱门形稍强回声,前方、左、右侧被上牙槽突强回声所包围;88.00%(88/100)的胎儿硬腭在3个位面上均可显示。32胎唇裂胎儿中,检出9胎合并腭裂并经随访证实,其腭裂部位、裂隙宽度、整体形态及走向显示清楚。结论应用三维超声观察胎儿硬腭简便易行。掌握正常胎儿硬腭三维超声声像图特征有助于提高胎儿腭裂的检出率及评判预后。     

孕早期超声观察腭线筛查胎儿唇腭裂

目的 评价孕早期超声观察腭线筛查胎儿唇腭裂的价值。方法 回顾性分析14 360胎接受超声颈后透明层厚度（NT）检查的孕早期胎儿,观察胎儿腭线表现,记录胎儿转归,评价孕早期超声观察腭线筛查胎儿唇腭裂的效能。结果 孕早期超声提示14 327胎（14 327/14 360,99.77%）腭线正常,其中7胎经随访证实存在唇腭裂;33胎（33/14 360,0.23%）腭线异常,其中4胎腭线为小裂隙,随访证实无唇腭裂,29胎随访证实腭线异常,包括小裂隙8胎、大裂隙4胎、前部缺失11胎及腭线变细/变短6胎。孕中期超声提示36胎唇腭裂,并于出生后或经引产证实,包括4胎单纯唇裂、10胎单纯继发性腭裂、17胎单侧唇腭裂,5胎双侧唇腭裂。超声观察腭线预测胎儿唇腭裂的敏感度为80.56%（29/36）,特异度为99.97%（14 320/14 324）,阳性预测值为87.88%（29/33）,阴性预测值为99.95%（14 320/14 327）。结论 孕早期超声观察胎儿NT平面腭线可作为筛查胎儿唇腭裂的指标,值得推广。     

矢状切面在妊娠早期超声筛查胎儿腭裂中的价值

目的探讨矢状切面在妊娠早期超声筛查胎儿腭裂中的价值。 方法回顾性选择2018年1月至2019年12月广东省妇幼保健院的31例妊娠早期腭裂的胎儿,分析头颈部超声检查矢状切面的异常征象,总结不同类型腭裂在矢状切面的超声表现以及其他结构异常情况。 结果（1）超声声像图表现：28例出现上颌骨间隙（90.3%,28/31）,其中14例为单侧腭裂,5例为双侧腭裂,9例为正中腭裂;27例表现为缺失“重叠线征”（87.1%,27/31）,其中正中腭裂5例,单侧腭裂15例,双侧腭裂6例,单纯腭裂1例;6例颌骨前突（19.4%,6/31）,均为双侧唇腭裂。（2）合并其他结构异常情况：妊娠早期及妊娠中期诊断腭裂病例中,合并胎儿结构异常分别占75.0%（15/20）、45.5%（5/11）,颈项透明层增厚分别占60.0%（12/20）、36.4%（4/11）。 结论矢状切面上颌间隙和缺失“重叠线征”是妊娠早期筛查胎儿腭裂的重要线索,颌骨前突是双侧腭裂的特征性超声表现。     

Fetal cleft lip and palate: sonographic diagnosis, chromosomal abnormalities, associated anomalies and postnatal outcome in 70 fetuses.

OBJECTIVE: The aim of this study was to determine the relationship between facial clefts, associated malformations and chromosomal abnormalities. STUDY DESIGN: Sonograms of 70 fetuses with cleft lip with or without cleft palate were prospectively and retrospectively evaluated in our tertiary referral center for the nature of the cleft lip or palate and for the nature of the associated anomalies. Additionally, karyotyping was performed in 63 of the 70 patients (90%). RESULTS: The frequency of additional anomalies and the mortality rate in this selected population varied with the type of cleft. None of the fetuses presenting an isolated cleft lip had additional anomalies and all survived. All fetuses presenting a median facial cleft had concurrent anomalies (particularly of the central nervous system (90%)) and a fatal outcome. Associated defects were more frequent in fetuses with bilateral clefts (72%) than in those with unilateral clefts (48%). Fetuses with a unilateral cleft lip with or without cleft palate had a better survival rate (52%) than those with a bilateral cleft lip with or without cleft palate (35%). The frequency and type of chromosomal abnormalities varied with the type of cleft. The highest rate of chromosomal abnormalities was found in fetuses with median clefts (82%). CONCLUSIONS: Although no conclusions regarding the prevalence of chromosomal or other anomalies in patients with a cleft lip with or without cleft palate in the general population could be drawn, the study revealed a strong relationship between the type of facial cleft, associated malformations, chromosomal abnormalities and fetal outcome.     

Prevalence of aneuploidy and additional anatomic abnormalities in fetuses and neonates with cleft lip with or without cleft palate: a population-based study in Utah.

OBJECTIVE: To determine the prevalence of aneuploidy and additional major anatomic abnormalities in fetuses and neonates with cleft lip with or without cleft palate. METHODS: All cases of cleft lip with or without cleft palate (cleft lip/cleft palate) occurring in Utah from 1995 through 1999 were reviewed by using the Utah Birth Defect Network population-based surveillance system. All pregnancy outcomes are included (stillborn, live born, and termination) in this analysis. RESULTS: Of 263 cases of cleft lip/cleft palate, 72 (27.4%) were unilateral cleft lip, 112 (42.6%) were unilateral cleft lip and cleft palate, 12 (4.6%) were bilateral cleft lip, and 67 (25.5%) were bilateral cleft lip and cleft palate. Fifteen (5.7%) of the 263 fetuses and neonates were aneuploid. One (1.2%) with cleft lip (unilateral and bilateral combined) was aneuploid. Five (4.5%) of the fetuses and neonates with unilateral cleft lip and cleft palate were aneuploid compared with 9 (13.4%) of fetuses and neonates with bilateral cleft lip and cleft palate. In known or presumed euploid fetuses and neonates, additional sonographically occult major anatomic abnormalities occurred in 5 (7.0%) of 71 with unilateral cleft lip, 18 (16.8%) of 107 with unilateral cleft lip and cleft palate, 1 (8.3%) of 12 with bilateral cleft lip, and 12 (20.7%) of 58 with bilateral cleft lip and cleft palate. These abnormalities primarily involved the heart and the central nervous system. CONCLUSIONS: Amniocentesis for karyotype should be offered in all cases of cleft lip/cleft palate because of the risk of aneuploidy. Patients should be counseled that sonographically occult additional anatomic abnormalities might be present with all clefts.     

Three-dimensional sonographic findings associated with ectrodactyly ectodermal dysplasia clefting syndrome.

Ectrodactyly-ectrodermal dysplasia-clefting (EEC) syndrome is a rare autosomal dominant genetic syndrome. This condition is characterized by varying degrees of ectrodactyly and syndactyly of the hands and feet, a cleft lip with or without a cleft palate, and ectodermal dysplasia. In addition, abnormalities of the genitourinary system occur frequently in association with this syndrome. The wide clinical variability of EEC syndrome has been well documented in the literature, and none of the 3 cardinal signs seem to be obligatory. We present a case of classic familial EEC syndrome diagnosed by prenatal sonography at 17 weeks' gestation. In addition to tetraectrodactyly and a unilateral cleft lip and palate, renal dysplasia was noted on serial sonographic evaluation. Three-dimensional (3D) surface rendering was used to supplement traditional 2-dimensional (2D) imaging to further evaluate the fetal phenotype.     

Small Deletions of SATB2 Cause Some of the Clinical Features of the 2q33.1 Microdeletion Syndrome

Recurrent deletions of 2q32q33 have recently been reported as a new microdeletion syndrome. Clinical features of this syndrome include severe mental retardation, growth retardation, dysmorphic features, thin and sparse hair, feeding difficulties and cleft or high palate. The commonly deleted region contains at least seven genes. Haploinsufficiency of one of these genes, SATB2, a DNA-binding protein that regulates gene expression, has been implicated as causative in the cleft or high palate of individuals with 2q32q33 microdeletion syndrome. In this study we describe three individuals with smaller microdeletions of this region, within 2q33.1. The deletions ranged in size from 173.1 kb to 185.2 kb and spanned part of SATB2. Review of clinical records showed similar clinical features among these individuals, including severe developmental delay and tooth abnormalities. Two of the individuals had behavioral problems. Only one of the subjects presented here had a cleft palate, suggesting reduced penetrance for this feature. Our results suggest that deletion of SATB2 is responsible for several of the clinical features associated with 2q32q33 microdeletion syndrome.     

Analysis of the candidate genes responsible for non-syndromic cleft lip and palate in Japanese people

In order to assess the association of alleles for candidate genes with non-syndromic cleft lip and palate, DNA samples from 43 Japanese patients were compared with those from 73 control subjects with respect to the genes encoding transforming growth factor alpha (TGFalpha), TGFbeta and gamma-aminobutyric acid type A receptor beta3 (GABRB3). The restriction fragment length polymorphisms of the 3'-non-coding region of the TGFalpha gene K-primer region were observed after digestion with NcoI and HinfI. Allele 4 was the most common among cases of cleft lip with or without cleft palate, whereas allele 2 was the most common among controls. A significant difference was found in this region between groups with cleft lip (with or without cleft palate) and controls (chi2=10.190; P=0.017). Three alleles of the TGFbeta2 gene were tested, and allele 2 was the most common in both cases and controls. The proportion of allele 2 in the case group was greater than that in the control group, showing a significant difference between cases of cleft lip (with or without cleft palate) and controls (chi(2)=19.208; P<0.0001). No significant differences in variants of TGFbeta3 or GABRB3 between case and control populations were observed. Thus it is concluded that TGF genes play a role in craniofacial development, and that alleles of TGFalpha or/and TGFbeta2 are associated with cleft lip and cleft palate in Japanese populations.     

二维超声及三维超声多种成像技术在胎儿唇腭裂畸形诊断中的联合应用

目的研究二维超声及三维超声多种成像技术在胎儿唇腭裂畸形诊断中的图像特征。 方法选取2016年1月至2017年12月在南京医科大学附属苏州医院产科接受中孕期结构畸形筛查发现唇腭部异常的胎儿均行三维超声表面成像、断层成像（TUI）及自由解剖成像（OmniView模式）多种成像技术联合检查,并随访至出生后或引产后。结合产前超声检查图像,研究唇腭裂胎儿颜面部三维超声图像特征。 结果产前超声筛查的19 168例中孕期胎儿中检出唇腭部结构畸形36例（0.19%）。36例唇腭裂胎儿超声征象：单纯唇裂（CL）8例,二维、TUI及OmniView模式均检出胎儿唇裂;三维表面成像漏诊1例唇红裂。二维及三维成像技术均显示胎儿上唇连续性中断,但三维成像显示更直观。唇裂合并上牙槽突裂（CLA）11例,二维超声显示8例,三维表面成像、TUI均诊断9例,漏诊2例,OmniView技术诊断10例,漏诊1例,联合检查诊断10例,漏诊1例。主要超声征像：上唇及上牙槽突连续性中断,二维超声对于牙槽突裂显示较困难,TUI及OmniView可从多角度显示牙槽突裂。唇裂合并腭裂（CLP）17例,二维超声（诊断10例）、三维超声表面成像模式（诊断13例）、TUI（诊断15例）、OmniView模式（诊断16例）均未全部检出;而17例CLP经联合检查全部检出。超声征象为上唇、原发腭及继发腭的连续中断,断层成像及OmniView可从多角度显示原发腭及继发腭,优于二维超声对于原发腭及继发腭的显示。 结论产前二维超声及三维超声能清晰显示胎儿唇裂,但对于腭裂,三维超声多种成像模式图像优于二维超声,产前超声筛查联合三维多成像技术能清晰显示胎儿颜面部异常,减少唇腭裂的漏诊。     

Prenatal detection of associated anomalies in fetuses diagnosed with cleft lip with or without cleft palate in utero.

OBJECTIVE: The aim of this study was to determine the prenatal detection rate of associated anomalies in fetuses with a suspected cleft lip with or without cleft palate. METHODS: Fetuses with a suspected cleft lip with or without cleft palate, determined by prenatal ultrasound, were prospectively enrolled. Additional anomalies suspected by ultrasound or genetic testing were recorded. Postnatal outcome was obtained. RESULTS: Forty-five fetuses with a cleft lip with or without cleft palate, diagnosed prenatally with either two-dimensional and/or three-dimensional ultrasound, were studied. Postnatal follow-up revealed that 16 (35.6%) of these 45 fetuses had an additional structural or syndromic abnormality. Of the 37 fetuses with prenatally determined 'isolated' cleft lip with or without cleft palate, eight (21.6%) had an additional malformation identified after delivery. CONCLUSION: In pregnancies complicated by a cleft lip with or without cleft palate, patients should be informed of the risks of associated anomalies, some of which may be undetected prenatally.