常染色体显性遗传性听神经病OTOF基因突变筛查

目的：了解OTOF基因在一常染色体显性遗传性听神经病家系的突变情况。方法：选择一个常染色体显性遗传听神经病家系中现存9名成员、3例散发听神经病患者和3名听力正常者为研究对象,用基因组DNA抽提试剂盒提取外周血DNA。对1例家系患者DNA进行OTOF基因全部编码区的PCR扩增,扩增产物经纯化后直接测序,测序结果与标准序列对照进行突变筛查;针对发现有突变的外显子,对其余受试者DNA样本进行PCR扩增和序列分析。结果：所有研究对象在OTOF基因相同部位上检测到10个新的碱基变异,但均未引起所编码氨基酸的改变。结论：该家系成员OTOF基因未发现有意义的突变位点,提示新基因参与家系耳聋的发生。     

常染色体显性遗传性听神经病基因突变筛查

目的针对一个常染色体显性遗传性非综合征性听神经病谱系障碍家系进行已知致病基因筛查分析,了解是否存在可能致病的基因突变。方法选择本课题组收集到的一个家系4代24人作为研究对象,采集受试者外周静脉血提取基因组DNA,应用在线引物设计软件Primers进行引物设计,采用直接测序法对已知听神经病相关基因进行测序,使用DNAMan软件对序列进行比对分析。结果该家系遗传方式符合显性遗传特征,患者听力学特征符合非综合征型听神经病谱系障碍,对家系中全部患者及部分听力正常成员完成DIAPH3、OTOF、PJVK、GJB2基因外显子及线粒体DNA12SrRNA1494、1555位点的测序分析,未发现可疑致病突变。结论该家系未检测到已知相关基因致病突变,高度提示新基因突变致病的可能,有待于进一步研究。     

常染色体显性遗传性听神经病DFNB59基因序列分析

目的:了解DFNB59基因是否与一个常染色体显性遗传性听神经病(AN)中国家系的发病相关。方法:以一个现存4代9人的AN家系为研究对象,用基因组DNA抽提试剂盒提取外周血DNA。对所有家系成员DNA进行DFNB59基因第2、第4外显子的PCR扩增,1例AN病患者进行DFNB59基因全部编码区的PCR扩增,扩增产物经纯化后直接测序,测序结果与标准序列对照进行突变位点鉴定。结果:所有研究对象的基因区域均扩增成功,序列分析在DFNB59基因第2、第4外显子上未检测到T54I和R183 W2个已知的突变,整个基因编码区也未发现新的致聋突变。结论:该家系成员DFNB59基因上未发现有意义的突变位点,提示新基因参与家系AN的发生。     

特大常染色体显性遗传聋家系听力学特征及候选致病基因突变筛查

目的探讨一中国常染色体显性遗传聋大家系的听力学特征,进行已知致聋基因已知突变位点的筛查。方法经知情同意,对家系成员进行全身检查及听力学检测,获得血样标本;整理分析家系资料并绘制系谱图;用基因组DNA抽提试剂盒提取外周血DNA。对2例家系患者DNA进行GJB2和GJB3基因全部编码区突变检测,对其余23个已知常染色体显性遗传性耳聋(DFNA)基因的74个已知突变位点所涉及的50个外显子进行PCR扩增和直接测序分析。结果该家系共7代199人,现存4代176人,耳聋患者54人。系谱分析显示,耳聋表型代代相传,男女患病人数分别为24和30,符合常染色体显性遗传特征。听力学表现为:迟发性、进行性、双侧对称性、感音神经性听力损失,首先是高频区受损,并快速向中、低频扩展。GJB2、GJB3基因全部编码区及其余23个DFNA基因已知突变位点的序列分析均无阳性发现。结论该家系是一个非综合征型常染色体显性遗传聋大家系,耳聋表型为迟发性、进行性、双耳对称性感音神经性听力损失;初步分子遗传学分析提示可能由新基因或已知基因的新突变致病。     

常染色体显性遗传性聋大家系的遗传学特征分析

目的 探讨遗传性聋基因的定位克隆研究。方法 对两个国人耳聋大家系的资料进行了收集、整理及临床遗传学特征的分析。通过先证者对家系成员进行调查并绘制系谱图。对调查的家系成员进行病史、体检、纯音测听、声导抗及听性脑干反应检查。一些家系成员进行了颞骨CT扫描检查以排除听觉系统的其他病变。结果 两个耳聋家系，其先证者均诊断为感音神经性聋，分别命名为Z1318及W727家系，表现为一种代代相传的中度至中重度听力损失。遗传方式为常染色体显性遗传，听力表型为一种迟发型的、渐进性的、以高频下降为主的听力损失，但两家系在听力下降的具体表现形式上存在差异。Z1318家系发病年龄各代间较稳定，为10-20岁，而W727家系各代差别较大，为8-30岁，且有逐代提前的趋势。Z1318家系以高频损失为主，听力曲线呈下降型；W727家系初以高频下降为主，随着年龄增长逐渐累及全频听力，听力曲线由下降型变为平坦型。结论 两个家系均为常染色体显性遗传方式，均适合于进一步的候选基因克隆及连锁分析、定位克隆研究，以便寻找到相应的耳聋相关基因。     

常染色体显性遗传性耳聋家系的遗传学特征分析

目的 听力损失是中国人群中的常见的感觉障碍性疾病。为了解遗传因素在中国听力损失病人中的作用,对两个中国耳聋大家系进行了遗传特征的分析。方法:家系中的先证者在解放军总医院耳鼻咽喉头颈外科就诊,诊断为感音神经性耳聋。通过先证者对家系成员进行调查并绘制系谱图。对调查的家系成员进行病史、体检、纯音测听及听性脑干诱发电位检查。一些家系成员进行了颞骨CT扫描检查以排除听觉系统的其他病变。结果:两个中国耳聋家系,命名为Z002及F013家系,表现为一种代代相传的中度及中重度听力掘失。遗传方式考虑为常染色体显性遗传方式。在Z002家系的听力表型表现为一种高频听力损失,而在F013家系表现为低频听力损失。结论:本文报道了两个特征为非综合征型的常染色体显性遗传的中国耳聋家系。系谱图分析提示两个家系均为常染色体显性遗传方式。这两个家系适合于进一步的连锁分析及定位克隆研究以便寻找到相应的耳聋相关基因。     

KCNQ4基因突变对常染色体显性遗传性聋家系的影响

目的 应用选基因法了解KCNQ4基因对中国耳聋家系的影响，检测其突变形式。方法 在一个6代相传的常染色体显性遗传性家系中，应用聚合酶链反应－单链构像多态性（polymerase chain reaction-single strand conformation polymorphism,PCR-SSCP)及克隆测序方法对KCNQ4基因的全部编码序列的PCR产物进行突变位点及多态序列检测。结果 在该家系中，对36位家系成员进行了KCNQ4基因的编码序列的检测，发现KCNQ4基因外显子2的分子多态现象，经测序分析证明这种多态是由于内含子中47个碱基复制数的差异所造成的。结论 本实验证明KCNQ4基因外显子2的编码区附近存在一个新的分子多态标记，这种分子多态表现出不同的基因型。通过对这些基因型与耳聋表型的相关分析发现，随着内含子复制数的增加，耳聋表现度明显增加。提示KCNQ4基因外显子2与外显子3之间内含子复制数的变化可能是这个家系出现耳聋的一种特征性分子标记。     

一个常染色体显性遗传性聋家系致病基因鉴定

目的分析常染色体显性遗传性聋家系的听力学及遗传学特征,利用高通量测序和连锁分析技术进行致病基因鉴定。方法采集一个常染色体显性遗传性非综合征型聋家系患者的临床资料,进行耳聋表型和遗传方式的判定并绘制家系图,提取家系成员外周血DNA,首先利用耳聋相关基因靶向测序,对家系先证者进行162个已知耳聋基因的筛查,然后采用全外显子组测序和连锁分析相结合的方法继续寻找致病基因,筛选出候选基因变异位点在家系中进行验证,以明确该家系致病原因。结果该耳聋家系来自河南省,编号为HBSY-012,现存三代共34人,14人诊断为感音神经性聋,为常染色体显性遗传模式,耳聋者发病年龄5~7岁,早期表现为高频听力下降,随年龄增长迅速发展为全频受累的重度或极重度感音神经性聋。对先证者进行已知162个耳聋基因筛查未发现致病突变,家系连锁分析将致病基因定位于第9号染色体q31.1-q31.3区间内(最大LOD值3.6076)。全外显子组测序数据分析显示在连锁分析定位的区间内未发现候选变异,在区间以外筛选出4个候选基因变异位点,候选变异为ANKMY2基因NM_020319c.822_826del、DDX49基因NM_019070c.341C>T、DEFB129基因NM_080831c.284G>T以及EVI5基因NM_005665c.2399C>T,并对4个候选基因变异位点进行家系验证,结果提示都不是该家系的致病突变。结论该常染色体显性遗传非综合征型聋家系连锁分析将致病基因定位于第9号染色体q31.1-q31.3区间内。耳聋相关基因靶向测序和全外显子组测序均未发现致病突变,考虑该家系致病原因可能为基因的非编码区域的突变或者罕见的CNV/SV所致。     

一个迟发性非综合征型常染色体显性遗传性聋家系表型特征及致病基因初步探讨

目的分析一个迟发性遗传性聋大家系的临床表型,探讨该家系耳聋患者的致病基因。方法对一个湖南籍耳聋大家系成员进行详细的病史资料采集、体格检查、听力学检查,其中两名患者做了颞骨CT检查。绘制家系图。以先证者外周血基因组DNA为模板对候选致病基因进行涵盖全部编码序列聚合酶链反应(polymer-ase chain reaction,PCR)扩增,对扩增产物进行酶切纯化后用ABI 3730测序仪直接测序,用DNASTAR-Laser-gene SeqMan Pro软件对测序结果进行分析。结果系谱分析得知该家系是一个常染色体显性遗传性非综合征型聋家系。患者临床表现高度一致,均表现为在9～25岁时首先出现"嗡嗡样"耳鸣,然后自觉双耳听力下降,纯音测听显示早期为以高频听力下降为主的神经性聋,之后听力下降程度逐步加重并波及低频。对候选致病基因进行突变检测,未发现致病突变。结论该家系符合常染色体显性遗传的特征,其致病基因还有待于进一步探索。     

Familial auditory neuropathy

OBJECTIVES/HYPOTHESIS: Auditory neuropathy is a sensorineural hearing disorder characterized by absent or abnormal auditory brainstem responses and normal cochlear outer hair cell function as measured by otoacoustic emission recordings. Many risk factors are thought to be involved in its etiology and pathophysiology. Four Chinese pedigrees with familial auditory neuropathy were presented to demonstrate involvement of genetic factors in the etiology of auditory neuropathy. STUDY DESIGN: Probands of the above-mentioned pedigrees, who had been diagnosed with auditory neuropathy, were evaluated and followed in the Department of Otolaryngology-Head and Neck Surgery, China People Liberation Army General Hospital (Beijing, China). Their family members were studied, and the pedigree maps established. METHODS: History of illness, physical examination, pure-tone audiometry, acoustic reflex, auditory brainstem responses, and transient evoked and distortion-product otoacoustic emissions were obtained from members of these families. Some subjects received vestibular caloric testing, computed tomography scan of the temporal bone, and electrocardiography to exclude other possible neuropathic disorders. RESULTS: In most affected patients, hearing loss of various degrees and speech discrimination difficulties started at 10 to 16 years of age. Their audiological evaluation showed absence of acoustic reflex and auditory brainstem responses. As expected in auditory neuropathy, these patients exhibited near-normal cochlear outer hair cell function as shown in distortion product otoacoustic emission recordings. Pure-tone audiometry revealed hearing loss ranging from mild to profound in these patients. Different inheritance patterns were observed in the four families. In Pedigree I, 7 male patients were identified among 43 family members, exhibiting an X-linked recessive pattern. Affected brothers were found in Pedigrees II and III, whereas in pedigree IV, two sisters were affected. All the patients were otherwise normal without evidence of peripheral neuropathy at the time of writing. CONCLUSION: Patients with characteristics of nonsyndromic hereditary auditory neuropathy were identified in one large and three smaller Chinese families. Pedigree analysis suggested an X-linked, recessive hereditary pattern in one pedigree and autosomal recessive inheritances in the other three pedigrees. The phenotypes in the study were typical of auditory neuropathy; they were transmitted in different inheritance patterns, indicating clinical and genetic heterogeneity of this disorder. The observed inheritance and clinical audiological findings are different from those previously described for nonsyndromic low-frequency sensorineural hearing loss. This information should facilitate future molecular linkage analyses and positional cloning for the relative genes contributing to auditory neuropathy.     

Autosomal recessive hereditary auditory neuropathy

Objectives: Auditory neuropathy (AN) is a sen-sorineural hearing disorder characterized by absent or abnormal auditory brainstem responses (ABRs) and normal cochlear outer hair cell function as measured by otoacoustic emissions (OAEs). Many risk factors are thought to be involved in its etiology and patho-physiology. Three Chinese pedigrees with familial AN are presented herein to demonstrate involvement of genetic factors in AN etiology. Methods: Probands of the above - mentioned pedigrees, who had been diagnosed with AN, were evaluated and followed up in the Department of Otolaryngology Head and Neck Surgery, China PLA General Hospital. Their family members were studied and the pedigree diagrams were established. History of illness, physical examination, pure tone audiometry, acoustic reflex, ABRs and transient evoked and distortion - product otoacoustic emissions (TEOAEs and DPOAEs) were obtained from members of these families. DPOAE changes under the influence of contralateral sound stimuli were obs     

小儿类听神经病的临床表现

目的:报道1组小儿类听神经病(ANSD)的临床特征,以加深对该病的认识。方法:回顾性分析84例(151耳)ANSD患儿病史、影像学、听力学表现及人工耳蜗植入效果。结果:①高胆红素血症11例,家族史2例,缺氧3例,早产1例,基底核病变1例,先天性脑发育落后2例,脑瘫1例,蜗神经发育不全13例;②79.8%(67/84)为双侧发病,20.2%(17/84)为单侧发病;③短声ABR:70.2%(106/151)在最大输出(100dB nHL)无反应波形引出,29.8%(45/151)在非常高的刺激强度仅有分化不良的Ⅴ波;④行为听力测试:23例有测试结果者中,1例为轻度感音神经性聋,2例重度,20例为极重度;⑤影像学:29例有内耳MRI者中,外、中、内耳结构正常者16例,蜗神经发育不良12例,蜗神经发育不良伴双脑室周围白质髓鞘形成不良1例;⑥人工耳蜗植入效果:5例患耳植入人工耳蜗,其中3例术后1年婴幼儿有意义听觉整合量表得分与蜗性聋儿相当,1例稍差于蜗性聋儿,另1例无效。结论:ANSD是听力学表现相同的一大类疾病,在病史、听力学、影像学及人工耳蜗植入效果方面存在多样性,因此对此类患儿应进行全面、详细的临床检查,以便制...     

Cochlear implantation in patients with auditory neuropathy of varied etiologies

OBJECTIVES/HYPOTHESIS: Auditory neuropathy is a relatively recently described pattern of hearing loss characterized by preservation of outer hair cell function despite absent brainstem auditory evoked responses. Intact outer hair cell function is demonstrated by the presence of otoacoustic emissions and/or a measurable cochlear microphonic on electrocochleography, whereas no synchronous neural activity (absent action potentials) is seen on acoustically evoked brainstem auditory evoked response testing. The study reviews the authors' experience with six patients diagnosed with auditory neuropathy, four of whom have undergone cochlear implantation. MATERIALS AND METHODS: A retrospective review of all medical and audiological charts at the University of Virginia Hospitals (Charlottesville, VA) was performed to identify patients who have undergone cochlear implantation or have been diagnosed with auditory neuropathy, or both. RESULTS: Six patients with hearing loss attributable to auditory neuropathy were identified, four of whom have undergone cochlear implantation. Causes varied, including congenital, infectious, and idiopathic origins. Adults demonstrated subjective auditory perception on promontory stimulation, whereas no repeatable brainstem auditory evoked response waveforms could be demonstrated on pediatric promontory stimulation testing. Patients with implants demonstrated implant-evoked brainstem auditory evoked responses and improved audiological performance. CONCLUSIONS: The six cases presented in the study represent varied causes and, probably, varied sites of lesions of auditory neuropathy. Promontory stimulation has been valuable, particularly in adults. Cochlear implantation allows the opportunity to provide a supraphysiological electrical stimulation to the auditory nerve, with the hope of reintroducing synchronous neural activity. Greater confidence and enthusiasm for cochlear implantation in appropriately selected patients with auditory neuropathy are gained through experience with such diverse cases.     

小儿听神经病的误诊和漏诊

目的:探讨小儿听神经病的临床表现。方法:对一组年龄为4～68个月,听力学检查表现为听神经病患儿的临床资料进行回顾性研究。受试者的入选标准为短声听觉脑干反应(ABR)严重异常而畸变产物耳声发射(DPOAE)和(或)耳蜗微音电位(CM)正常。分析其短声ABR、DPOAE、行为听力测试、声导纳以及内耳MRI等结果。结果:①40例听力学表现为双侧听神经病,8例为单侧;②68.2%耳(60/88)在短声ABR的最大输出强度100dBnHL无反应,而有些耳仅在很高的测试强度有波形分化较差的Ⅴ波;③所有耳均可记录到CM,而只有41.5%能记录到DPOAE;④该组患儿的行为听力结果各种各样,为从轻度到极重度的听力损失;⑤4例经内耳MRI诊断为蜗神经发育不全。结论:在听力学表现为听神经病的儿童:①CM的诊断敏感度较DPOAE高;②各种客观听力测试无法预估行为听力;③需作内耳MRI检查,以除外蜗神经发育不全。     

听神经病谱系障碍患者DIAPH3基因临床遗传学特征研究

目的对中国听神经病谱系障碍患者进行DIAPH3基因的筛查,了解中国听神经病谱系障碍人群中是否存在DIAPH3基因的新突变及热点突变位点,并结合临床表型与基因型进行研究,探讨DIAPH3基因相关的听神经病谱系障碍患者的临床遗传学特征。方法选取2003—2013年我院确诊的并接受血样采集的125例听神经病谱系障碍患者,同时选取100例听力正常的健康志愿者作为对照。采集外周静脉血,提取基因组DNA,采用直接测序法进行DIAPH3基因突变的检测,选用DNAStar软件和Sequence Scanner软件进行序列比对分析。同时分析患者的临床特征,从临床表型与基因型两方面进行系统研究,探讨DIAPH3基因相关的听神经病谱系障碍患者的临床遗传学特征。结果在125例听神经病谱系障碍患者中,共发现DIAPH3基因外显子区域的22处变异,包括可能致病突变7处(c.1425G>A,c.1749T>C,c.2605A>C,c.-44C>G,c.-179G>A,c.-27C>T和c.*196A>T)。结论此7种突变均于中国听神经病谱系障碍人群中发现,在国内外尚未见报道,可能为听神经病谱系障碍相关的突变,有待于进一步的研究。