核心家系孤独症患儿FOXP1基因的突变及CNV分析

目的 研究转录因子FOXP1在汉族孤独症患儿核心家系中的外显子突变,并进行初步的功能预测。 方法 288例孤独症患儿及生物学父母进行FOXP1外显子区域基因测序和拷贝数变异(copy number variation,CNV)检测;使用POLYPHEN及SIFT等软件进行功能预测。 结果 4例患儿的FOXP1基因外显子区域发现4个错义突变,包括P42S,H53Q,L68R和M590V。在CNV检测中为阴性结果。通过功能预测发现,FOXP1的四个罕见突变可能不会对FOXP1编码蛋白产生重要影响。 结论 汉族人群中,FOXP1基因可能不是孤独症的主要致病基因。     

转录因子ZIC3和GATA4突变与先天性心脏病的关系

目的探讨锌指转录因子3 (ZIC3)和GATA4突变与先天性心脏病(CHD)的关系。方法选取2016年1月-2018年1月该院住院部收治的188例CHD患者作为研究组,180例体检健康者作为对照组,收集受试者外周血,提取DNA,进行PCR扩增及DNA测序分析。结果 ZIC3测序显示,8例房间隔缺损患者发生杂合错义突变c. 1341AG,位于外显子3; 4例房间隔缺损患者发生杂合错义突变c. 1154CG,位于外显子1,2例室间隔缺损患者发生杂合错义突变c. 890GT,位于外显子2; GATA4测序显示,25例室间隔缺损患者发生杂合同义突变c. 744CT,位于外显子3,13例发生杂合错义突变c. 1220CA,位于外显子4; 7例房间隔缺损发生杂合同义突变c. 839CT,位于外显子3; 3例家族三代房间隔缺损出现错义突变c. 899AC,位于外显子4; 2例房间隔缺损患者发生杂合错义突变c. 1138GA,位于外显子4; 1例动脉导管未闭患者发生新的杂合同义突变c. 685CT,位于外显子2。结论 CHD患者中ZIC3外显子发现3个突变位点,GATA4发现6个突变位点,ZIC3和GATA4突变基因可能与部分CHD致病相关。     

孤独症谱系障碍儿童的磁共振弥散张量成像研究

目的 探讨孤独症谱系障碍儿童重要区域脑白质的磁共振弥散张量成像(diffusion tensor imaging,DTI)特点。方法 对37例男性孤独症谱系障碍儿童和16例正常发育男童进行神经心理评估及磁共振弥散张量成像技术扫描,比较两组各向异性分数(fractional anisotropy,FA)的差异,并分析孤独症组感兴趣区FA值与儿童智能发展水平及临床表现之间的关系。结果 孤独症谱系障碍组的感兴趣脑区FA值与对照组相比无明显差异(P>0.05)。孤独症谱系障碍组的左侧额叶FA值与智能发展水平有相关性(P<0.05)。孤独症谱系障碍儿童儿童期孤独症评定量表(The Childhood Autism Raling Scale,CARS)分级重度组比轻-中度组左侧额叶FA值更低(P<0.05)。结论 左侧额叶白质损伤可能参与了孤独症谱系障碍的病例生理过程的变化。     

帕金森病Parkin,LRRK 2基因序列变异研究

目的 探讨散发性早发性帕金森病患者遗传易感基因突变的形式和分布及易感基因突变在PD发病中的可能作用.方法 病例组由23例散发性早发帕金森患者组成,10例对照组.以基因组DNA为模板,扩增Parkin基因的第1、4、6号外显子和LRRK 2基因的第31号外显子.观察PCR产物测序后的突变情况.结果 发现样本中存在突变以及单核苷酸多态性(Single Nucleotide Polymorphism,SNP).在一例患者Parkin基因Exon6上发现735ntT→C,对应的密码子TGT212CGT,翻译的氨基酸C212R,国内罕有报道.另一例患者Parkin基因Exon6上发现突变833ntG→C,导致第244密码子同义突变.结论 Parkin基因外显子4、6的突变可能是我国散发性早发PD患者的致病原因之一.     

婴幼儿尘螨严重过敏症的遗传学初步研究

目的 对14例0岁～4岁婴幼儿尘螨严重过敏症进行全外显子组测序，探讨其可能的遗传学致病因素。方法 对14例患儿进行全外显子组测序，对相关变异位点进行Sanger测序验证。通过文献及数据库查找，筛选与过敏性疾病相关的基因集，并在外显子组水平、疾病相关基因集水平比较患者组和对照组的变异负荷。结果 通过对受检者进行遗传病相关基因测序、生物信息学分析、遗传分析(遗传模式包括常染色体显性遗传、常染色体隐性遗传、X/Y连锁)及结合临床表型的综合分析，未检测到与受检者临床表型相关的已知或可能具有临床意义的基因变异。与对照组相比，患者组在外显子组及2个过敏性疾病相关基因集范围内的功能缺失性变异和罕见非同义有害变异负荷均无显著差异。结论 过敏性疾病有明显遗传倾向，但全外显子测序结果显示未发现重复的变异基因，使得对于和过敏有关的基因需要进一步深入细致的研究。     

海口市学龄前儿童孤独症谱系障碍筛查率及其影响因素

目的 了解海口市学龄前儿童孤独症筛查阳性率及其影响因素。方法 采取分层随机整群抽样的方法,抽取海口市23家幼儿园学龄前儿童共3 844名,使用克氏孤独症量表和自制的影响因素调查问卷进行调查。结果 本次调查的3 844例儿童,年龄分布在24~84个月,男女比例为1.26〖DK〗∶1,城市与农村比例为2.06〖DK〗∶1,汉族与少数民族比例为25.69〖DK〗∶1。孤独症谱系障碍筛查阳性率为5.10%,单因素分析发现,男性、低年龄、农村户籍、父母文化程度低、低出生体重、宫内发育迟缓、高龄产妇组儿童的孤独症谱系障碍筛查阳性率较高,差异有统计学意义。多重线性回归分析发现,父母文化程度高、城市户籍为孤独症谱系障碍的保护因素,男童、低年龄、低出生体重、宫内发育迟缓为孤独症谱系障碍的危险因素。结论 应该以男童、低年龄、农村户籍、有低出生体重、宫内发育迟缓史的学龄前儿童为重点观察对象,加强家长以及幼师对孤独症谱系障碍的认识,做到早期识别,早诊断、早干预。     

无精子症患者基因检测分析

目的:利用全外显子测序技术,筛选无精子症患者相关基因,丰富男性不育基因库。方法:抽取20例无精子症患者外周血,提取DNA,使用杂交捕获方法构建DNA文库,采用高通量测序技术检测人类全外显子组中20099个基因的外显子区及旁侧内含子区(20bp),将测序数据与人类基因组hg19参考序列进行比对,筛选变异基因,变异位点进行Sanger测序验证。结果:共计筛选出26个基因43个变异位点,排除15个常染色体隐性遗传的单个变异位点及13个与精子运动相关的变异位点,剩余11个基因的15个变异位点可能与精子发生障碍相关,其中包括FAM71B、STARD9、CLTCL1、PCBP3、S100PBP等5个在睾丸组织高表达基因,SYCE3、EFCAB6、DDX4、KDM5D、RGS22、MTL5等6个基因可能与精子发生障碍相关。结论:通过研究筛选到可能影响男性不育的基因,为男性不育的基因诊断研究提供参考。     

孤独症谱系障碍儿童父母性格特征和家庭环境的病例对照研究

探讨孤独症谱系障碍(ASD)儿童父母性格特征和家庭环境特点及其与儿童孤独症谱系障碍发病的关系,为预防由于父母性格问题导致的孤独症儿童养育环境不良因素发生提供参考.方法 应用家庭环境量表中文版(FES-CV)和成人版艾森克人格问卷(EPQ)对79例孤独症谱系障碍患儿和79例正常儿童的父母进行对照研究.结果 ASD组儿童父亲掩饰程度、母亲内外向、家庭亲密度、情感表达、知识性、娱乐性、道德宗教观、组织性和控制性得分均低于正常组(t值分别为-2.95,-2.33,-2.50,-3.15,-3.41,-3.15,-3.92,-3.42,P值均<0.05);病例组儿童家庭矛盾性、父亲神经质和母亲神经质得分高于正常组(t值分别为-2.50,2.29,2.23,P值均<0.05).回归分析显示,家庭矛盾性、知识性、组织性、控制性、父亲掩饰程度、母亲内外向是孤独症谱系障碍发病的影响因素(P值均<0.05).结论 孤独症谱系障碍患儿父母性格偏离,家庭环境不良,且与患儿发病有关.应重视父母的心理健康及家庭环境的调整.     

汉族孤独症谱系障碍儿童脆性X基因突变研究

【目的】 采用优化的PCR体系对脆性X智力障碍基因-1(FMR-1)片段扩增,分析汉族孤独症谱系障碍(autism spectrum disorder, ASD)患儿FMR-1基因异常突变发生情况。 【方法】 以466例汉族ASD患儿为研究对象,其中典型孤独症433名,未分类广泛性发育障碍(PDD-NOS)33名。取患儿外周血3~5 mL提取DNA,采用优化的PCR体系对466例患儿DNA进行FMR-1基因片段扩增;对于扩增成功的PCR产物进一步采用短串联重复序列(short tanderm repeat, STR)荧光检测技术进行检测。 【结果】 466个ASD患儿中,464个ASD患者(99.57%)DNA样品PCR扩增成功;2个典型孤独症患者(0.43%)DNA样品PCR扩增失败(排除DNA质量和操作问题),推断FMR-1基因发生异常突变。对于扩增成功的PCR产物进一步采用STR荧光检测技术进行检测,结果显示PCR产物均小于500 bp,其(CGG)n重复个数属正常突变范围。 【结论】 研究采用优化的PCR体系对FMR-1基因片段扩增,进一步采用STR荧光检测技术准确推断出基因片段中三核苷酸(CGG)重复个数,优化后的PCR体系可适用于大样本孤独症谱系障碍儿童的FMR-1基因突变检测。检测结果显示汉族孤独症谱系障碍儿童FMR-1基因异常突变率为0.43%。     

孤独症谱系障碍病因影响因素分析

目的探讨孤独症谱系障碍发病的影响因素,为孤独症谱系障碍的病因、发病机制及防治提供依据。方法采用自制的孤独症谱系障碍影响因素调查表对80例孤独症谱系障碍患儿和80例正常儿童进行病例对照研究。结果经非条件多因素回归分析结果显示,有统计学意义的各个危险因素中,对孤独症谱系障碍发病的影响值从大到小依次为:过期产(OR=20.396,P0.01),人工喂养(OR=7.567,P0.01),新生儿黄疸(OR=7.234,P0.01),宫内或新生儿缺氧(OR=6.411,P0.05),孕期接触不良因素(OR=3.354,P0.01),塑料玩具(OR=3.115,P0.01),母亲性格内向(OR=2.086,P0.05),而常规的家庭教育为保护因素(OR=0.093,P0.01)。结论孤独症谱系障碍儿童的发病可能与遗传,环境因素,孕期和出生高危因素存在一定关系,控制这些因素可能会降低孤独症谱系障碍的发生。     

三种人全外显子组捕获探针的性能比较

目的比较不同平台3个全外显子组探针的性能,为以后建立标准流程提供数据支撑。 方法提取人EDTA全血或FFPE组织gDNA,通过酶切方法将gDNA片段化,并为每一个样本的片段化后的DNA加上特异序列的接头,利用IDT、Human Exome和MedExomed三种不同的液相探针的方法将目标基因捕获出来,用Illumina的NextSeq 500二代测序仪进行测序;分析测序数据,比较不同捕获探针的靶向覆盖率、捕获特异性、变异检出能力等。 结果本研究所选入组的3种探针,IDT和MedExome展示了对CCDS和Refseq数据库很好的覆盖率(均>95%);都表现了对靶区域很高的覆盖率(均>99%);IDT探针对血样gDNA捕获特异性(76.96%±0.75%,n=6),明显高于Human Exome (67.63%±1.62%,n=6) (P<0.001);IDT对FFPE标准品捕获特异性为84.48%±0.64%(n=3),明显高于MedExome的71.07%±0.91%(n=3)(P<0.01)。变异输出数量上,Human Exome最高。 结论IDT探针的捕获特异性明显高于其它2种,而在变异输出数量上,Human Exome探针最高。     

Joint genotype calling with array and sequence data

Analysis of rare variants is currently a major focus of genetic studies of human disease. Single-nucleotide polymorphism (SNP) genotypes can be assayed using microarray genotyping or by sequencing, but neither technology produces perfect genotype calls, especially at rare SNPs. Studies that collect both types of data are becoming increasingly common, so it may be possible to combine data types to increase accuracy. We present a method, called Chiamante, which calls genotypes on individuals with either array data, sequence data, or both. The model adapts to data quality and can estimate when either the array or the sequence data should be ignored when calling the genotypes at each SNP. As a special case, our method will call genotypes from only array data and outperforms existing methods in this scenario. We have applied our method to array and sequence data from Phase I of the 1000 Genomes Project and show that it provides improved performance, especially at rare SNPs. This method provides a foundation for future efforts to fuse genetic data from different sources, for example, when combining data from exome sequencing and exome microarrays.     

Parent-reported prevalence of autism spectrum disorders in US-born children: an assessment of changes within birth cohorts from the 2003 to the 2007 National Survey of Children's Health

The prevalence of autism spectrum disorders (ASD) from the 2007 National Survey of Children's Health (NSCH) was twice the 2003 NSCH estimate for autism. From each NSCH, we selected children born in the US from 1990 to 2000. We estimated autism prevalence within each 1-year birth cohort to hold genetic and non-genetic prenatal factors constant. Prevalence differences across surveys thus reflect survey measurement changes and/or external identification effects. In 2003, parents were asked whether their child was ever diagnosed with autism. In 2007, parents were asked whether their child was ever diagnosed with an ASD and whether s/he currently had an ASD. For the 1997-2000 birth cohorts (children aged 3-6 years in 2003 and 7-10 years in 2007), relative increases between 2003 autism estimates and 2007 ASD estimates were 200-600 %. For the 1990-1996 birth cohorts (children aged 7-13 years in 2003) increases were lower; nonetheless, differences between 2003 estimates and 2007 "ever ASD" estimates were >100 % for 6 cohorts and differences between 2003 estimates and 2007 "current ASD" estimates were >80 % for 3 cohorts. The magnitude of most birth cohort-specific differences suggests continuing diagnosis of children in the community played a sizable role in the 2003-2007 ASD prevalence increase. While some increase was expected for 1997-2000 cohorts, because some children have later diagnoses coinciding with school entry, increases were also observed for children ages ≥ 7 years in 2003. Given past ASD subtype studies, the 2003 "autism" question might have missed a modest amount (≤ 33 %) of ASDs other than autistic disorder.     

The Actionability of Exome sequencing testing results

Genomic tests such as exome sequencing have recently become an option for diagnosing patients. The tests allow clinical geneticists to sequence the majority of patients’ disease causing genetic variants. As a new technology, exome sequencing confronts the question of what the benefit is of this increased genetic information. Against a narrow perspective of clinical utility that emphasises tangible improvements in a patient's disease management, professional organisations have argued that genomic sequencing should be considered beneficial if it helps families and society. Based on video‐recorded observations of the return of exome sequencing results to parents of a child with disabilities in the clinic and in‐depth interviews with these parents, we examine how genomic test results become actionable in the clinical encounter. We find that parents and clinicians marshal exome results beyond biomedical diagnostic and management goals to address questions about guilt for causing the disabilities and to secure access to disability‐related services. We argue that genomic actionability rests on the interaction between the biological characteristics of genetic results and the predicaments facing parents of children with disabilities.     

孤独症谱系障碍儿童肠道菌群多样性的研究

目的 分析孤独症谱系障碍(ASD)儿童肠道菌群多样性,为减轻胃肠道症状提供科学依据。方法 选取2018年1-6月在广西壮族自治区妇幼保健院确诊为ASD的儿童20例和健康儿童20例作为研究对象。采集两组儿童粪便提取总DNA,扩增16S rDNA中V4/V5区进行高通量测序,分析ASD儿童肠道菌群多样性。结果 两组在样品文库的覆盖率(Z=242.500,P=0.256)、丰度指标Chao1(Z=250.000,P=0.181)、 ACE(Z=234.000,P=0.365)、Shannon(Z=259.000,P=0.114)和Simpson(Z=146.000,P=0.145)指数上差异均无统计学意义(P>0.05),多样性无差异。门水平上,两组儿童在8种菌群上差异无统计学意义(P>0.05),属水平上,两组儿童在20个属上差异具有统计学意义(P＜0.05)。结论 ASD儿童和健康儿童在肠道菌群属水平上具有差异;临床上可根据不同菌属特点对ASD儿童肠道菌群进行精准调整,保持肠道微生态平衡,减轻ASD儿童胃肠道症状。     

孤独症谱系障碍儿童肠道菌群多样性的研究

目的 分析孤独症谱系障碍(ASD)儿童肠道菌群多样性,为减轻胃肠道症状提供科学依据。方法 选取2018年1-6月在广西壮族自治区妇幼保健院确诊为ASD的儿童20例和健康儿童20例作为研究对象。采集两组儿童粪便提取总DNA,扩增16S rDNA中V4/V5区进行高通量测序,分析ASD儿童肠道菌群多样性。结果 两组在样品文库的覆盖率(Z=242.500,P=0.256)、丰度指标Chao1(Z=250.000,P=0.181)、 ACE(Z=234.000,P=0.365)、Shannon(Z=259.000,P=0.114)和Simpson(Z=146.000,P=0.145)指数上差异均无统计学意义(P>0.05),多样性无差异。门水平上,两组儿童在8种菌群上差异无统计学意义(P>0.05),属水平上,两组儿童在20个属上差异具有统计学意义(P＜0.05)。结论 ASD儿童和健康儿童在肠道菌群属水平上具有差异;临床上可根据不同菌属特点对ASD儿童肠道菌群进行精准调整,保持肠道微生态平衡,减轻ASD儿童胃肠道症状。     

Parent-Reported Prevalence of Autism Spectrum Disorders in US-Born Children: An Assessment of Changes within Birth Cohorts from the 2003 to the 2007 National Survey of Children’s Health

The prevalence of autism spectrum disorders (ASD) from the 2007 National Survey of Children’s Health (NSCH) was twice the 2003 NSCH estimate for autism. From each NSCH, we selected children born in the US from 1990 to 2000. We estimated autism prevalence within each 1-year birth cohort to hold genetic and non-genetic prenatal factors constant. Prevalence differences across surveys thus reflect survey measurement changes and/or external identification effects. In 2003, parents were asked whether their child was ever diagnosed with autism. In 2007, parents were asked whether their child was ever diagnosed with an ASD and whether s/he currently had an ASD. For the 1997–2000 birth cohorts (children aged 3–6 years in 2003 and 7–10 years in 2007), relative increases between 2003 autism estimates and 2007 ASD estimates were 200–600 %. For the 1990–1996 birth cohorts (children aged 7–13 years in 2003) increases were lower; nonetheless, differences between 2003 estimates and 2007 “ever ASD” estimates were >100 % for 6 cohorts and differences between 2003 estimates and 2007 “current ASD” estimates were >80 % for 3 cohorts. The magnitude of most birth cohort-specific differences suggests continuing diagnosis of children in the community played a sizable role in the 2003–2007 ASD prevalence increase. While some increase was expected for 1997–2000 cohorts, because some children have later diagnoses coinciding with school entry, increases were also observed for children ages ≥7 years in 2003. Given past ASD subtype studies, the 2003 “autism” question might have missed a modest amount (≤33 %) of ASDs other than autistic disorder.     

Prevalence of autism spectrum disorder among Nigerian children with intellectual disability: a stopgap assessment

The prevalence of autism spectrum disorder (ASD) among sub-Saharan African children with intellectual disability is about 0.7% more than three decades ago. During this period, the prevalence of ASD has been on the increase worldwide. Studies are not available in recent times addressing epidemiology of ASD among sub-Saharan African children. The present study assessed the prevalence of ASD among Nigerian children with intellectual disability. Forty four children with intellectual disability were assessed for diagnosis of childhood autism based on criteria specified in F84.0 section of International Classification of Diseases, Tenth Edition (ICD-10) Diagnostic Criteria for Research. Five (11.4%) of the children studied met the diagnostic criteria for childhood autism. Male/female ratio was 4:1. There is need for large scale epidemiological studies of ASD among sub-Saharan African children to clearly define the inter-relationship between ASD and intellectual disability in this population and to help in health care policy formulation.     

Factors associated with caregiving burden and maternal pessimism in mothers of adolescents with an autism spectrum disorder in Taiwan

Relative to the United States and other western countries, less research has focused on factors associated with caregiving burden and maternal pessimism in Taiwanese mothers of adolescents with an autism spectrum disorder (ASD). The characteristics of 50 adolescents with an ASD living at home in Taiwan and its association with caregiving burden and maternal pessimism were examined. The age range of adolescents with an ASD was from 10 to 18. Mothers, aged 35 to 55 years, completed self-report written questionnaires regarding their child's adaptive functioning and their own perceptions of caregiving burdens and concerns. Findings indicated that functional independence, severe maladaptive behaviours and severity of autism were predictive of maternal caregiving burden. Maternal pessimism was associated with functional independence and severity of autism. The findings of this study indicated that occupational therapy practitioners could focus on training functional independence of the individual with an ASD to meet the family's need in Taiwan. Researchers should pay significant attention to the lifespan issues of autism in Taiwanese families. The major limitations of this study were small sample size and without a comparison group. Future research using larger samples with a comparison group is needed.