[目的]探讨POU4F3基因单核苷酸多态性（SNPs）与中国人群中噪声性高频听力损失易感性之间的关系。[方法]采用1∶1配对病例对照研究方法,依据听力测试结果,将双耳高频平均听阈≥40 d B定义为病例组,双耳语频的任意频段和高频平均听阈均〈...[目的]探讨POU4F3基因单核苷酸多态性（SNPs）与中国人群中噪声性高频听力损失易感性之间的关系。[方法]采用1∶1配对病例对照研究方法,依据听力测试结果,将双耳高频平均听阈≥40 d B定义为病例组,双耳语频的任意频段和高频平均听阈均〈25 d B定义为对照组,共248对。对噪声暴露作业工人进行健康检查、问卷调查和纯音听力测试。利用SNPscanTM多重SNP分型试剂盒检测rs1368402和rs891969位点的基因型。分别采用配对t检验和配对χ2检验比较两组间计量资料和计数资料的分布差异,利用条件logistic回归模型分析2个SNP位点与噪声性高频听力损失易感性之间的关联。[结果]经吸烟、饮酒、高血压和累计噪声暴露量（CNE）校正后,POU4F3基因rs1368402和rs891969位点的基因型和等位基因分布在病例组与对照组之间的差异无统计学意义（P〉0.05）。按CNE分层后发现,当CNE〉95 d B（A）时,与rs1368402位点CC/CA基因型相比,携带AA基因型的个体发生噪声性高频听力损失的风险增加（校正OR=1.547,95%CI=1.002~2.389,P〈0.05）;与rs891969位点AA/GA基因型相比,携带GG基因型的个体发生噪声性高频听力损失的风险增加（校正OR=1.650,95%CI=1.032~2.639,P〈0.05）。并按噪声暴露水平、吸烟、饮酒和高血压进行分层分析,未见有统计学意义的结果（P〉0.05）。单体型分析中也未见病例组与对照组单体型频率差异有统计学意义（P〉0.05）。[结论]POU4F3基因单个位点多态性可能不是影响噪声性高频听力损失的遗传易感性因素,但rs1368402和rs891969位点多态性与噪声之间的交互作用可能对其具有重要影响。经Bonferroni检验校正后,上述相关性不再具有统计学意义,研究结果还需进一步验证。展开更多
Objective:The zebrafish is an excellent model for studying gene function in auditory system development.Pou4f3 plays an important role in mouse hair cell formation.Here,we constructed apou4f3-knockoutTg(Brn3c:GFP)zebr...Objective:The zebrafish is an excellent model for studying gene function in auditory system development.Pou4f3 plays an important role in mouse hair cell formation.Here,we constructed apou4f3-knockoutTg(Brn3c:GFP)zebrafish to provide an efficient fluorescence-visualized model for studying the molecular mechanisms of ear development.Methods:Cas9/single guide RNAs targeting exon 2 ofpou4f3 were designed and injected into one-cell stage zebrafish embryos(G0 generation).The G0 generation were crossed withTg(Brn3c:GFP)zebrafish to obtainpou4f3-mutantTg(Brn3c:GFP)zebrafish.The targeting efficiency was detected by polymerase chain reaction amplification and Sanger sequencing.Zebrafish hair cells were observed by laser scanning confocal microscopyin vivo.The morphology of the otoliths and semicircular canals were analyzed.All animal experiments were approved by the Animal Care and Use Committee of Shandong Provincial Hospital,Cheeloo College of Medicine,Shandong University(approval No.2016-KY-040)on March 3,2016.Results:Thepou4f3-mutantTg(Brn3c:GFP)zebrafish line was successfully established.Fluorescence observation suggested that hair cell development was delayed inpou4f3-knockout zebrafish.Knockout ofpou4f3 also induced defects in the otoliths and semicircular canals and impaired ear function in zebrafish.Conclusion:A CRISPR/Cas9-mediatedpou4f3 mutantTg(Brn3c:GFP)zebrafish model was established for the first time to demonstrate the essential role ofpou4f3 in zebrafish ear development.Our study provides a highly efficient method for the establishment of a visualized model of gene knockout zebrafish and has the potential to allow high-throughput drug screening to explore therapeutics for related diseases.展开更多
Objective： Based on the clinical manifestations of a hearing loss patient, the POU3F4 gene was tested for diagnosis of etiology. Methods： A comprehensive physical examination was performed on the proband to exclude ...Objective： Based on the clinical manifestations of a hearing loss patient, the POU3F4 gene was tested for diagnosis of etiology. Methods： A comprehensive physical examination was performed on the proband to exclude abnormalities of other organs, and detailed audi- ological testing and temporal bone CT scan were also performed. Genomic DNA was extracted using the proband＇s peripheral blood leukocytes. Polymerase chain reactions （PCR） were performed in the coding sequence of the POU3F4 gene. Direct DNA sequencing was subsequently applied to screen the entire coding region of the POU3F4 gene. Results： The proband had severe sensorineural hearing loss. Temporal CT showed bilateral cochlear incomplete partition, vestibule dysplasia, internal auditory canal fundus expansion, and cochlear interlink with the internal auditory canal fundus. A novel mutation （c.530C 〉 A （p.S 177X）） in the POU3F4 gene was found in this patient, creating an new stop codon and was predicted to result in a truncated protein lacking normal POU3F4 transcription factor function. Conclusion： Through analysis of the POU3F4 gene and clinical manifestations in the patient, we conclude that a novel mutation may have resulted in a premature stop codon, contributing to the mutation of POU3F4 gene.展开更多
We present the clinical and genetic findings for a Chinese family with X-linked non-syndromic hearing loss in which the affected males showed congenital profound sensorineural hearing impairment. In two affected broth...We present the clinical and genetic findings for a Chinese family with X-linked non-syndromic hearing loss in which the affected males showed congenital profound sensorineural hearing impairment. In two affected brothers, the computer tomography of temporal bone showed bilateral dilation of the internal auditory canal with fistulous communication between the lateral canal and the basal cochlear turn, which is consistent with the typical DFNX2 phenotype. A missense mutation （c.647G→A） in the POU3F4 gene caused a substitu- tion from glycine to glutamic acid at position 216 （p.G216E）, and this mutation was found to consistently cosegregate with the deafness phenotype in the family. The mutation resulted in the loss of function of the POU3F4 by decreasing the affinity between the protein and DNA, as shown in silico by the structural analysis. Prenatal diagnosis of pregnant proband of this family revealed the c.647G→A mutation in DNA extracted from the amniotic fluid surrounding the fetus. The appropriate use of genetic testing and prenatal diagnosis plays a key role in reducing the recurrence of genetic defects in high-risk families.展开更多
Background： The molecular genetic research showed the association between X-linked bearing loss and mutations in POU3F4. This research aimed to identify a POU3F4 mutation in a nonsyndromic X-linked recessive hearing ...Background： The molecular genetic research showed the association between X-linked bearing loss and mutations in POU3F4. This research aimed to identify a POU3F4 mutation in a nonsyndromic X-linked recessive hearing loss family. Methods： A series of clinical evaluations including medical history, otologic examinations, l：amily history, audiologic testing, and a high-resolution computed tomography scan were performed t＇or each patient. Bidirectional sequencing was carried out for all polymerase chain reaction products or＇the samples. Moreover, 834 controls with normal hearing were also tested. Results： The pedigree showed X-linkage recessive inheritance pattern, and pathogenic mutation （c.499C〉T） was identified in the proband and his family member, which led to a premature termination prior to the entire POU domains. This mutation co-segregated with bearing loss in this family. No mutation ofPOU3F4 gene was found in 834 controls. Conclusions： A nonsense mutation is identified in a family displaying the pedigree consistent with X-linked recessive pattern in POU3F4 gene. In addition, we may provide molecular diagnosis and genetic counseling for this family.展开更多
文摘[目的]探讨POU4F3基因单核苷酸多态性（SNPs）与中国人群中噪声性高频听力损失易感性之间的关系。[方法]采用1∶1配对病例对照研究方法,依据听力测试结果,将双耳高频平均听阈≥40 d B定义为病例组,双耳语频的任意频段和高频平均听阈均〈25 d B定义为对照组,共248对。对噪声暴露作业工人进行健康检查、问卷调查和纯音听力测试。利用SNPscanTM多重SNP分型试剂盒检测rs1368402和rs891969位点的基因型。分别采用配对t检验和配对χ2检验比较两组间计量资料和计数资料的分布差异,利用条件logistic回归模型分析2个SNP位点与噪声性高频听力损失易感性之间的关联。[结果]经吸烟、饮酒、高血压和累计噪声暴露量（CNE）校正后,POU4F3基因rs1368402和rs891969位点的基因型和等位基因分布在病例组与对照组之间的差异无统计学意义（P〉0.05）。按CNE分层后发现,当CNE〉95 d B（A）时,与rs1368402位点CC/CA基因型相比,携带AA基因型的个体发生噪声性高频听力损失的风险增加（校正OR=1.547,95%CI=1.002~2.389,P〈0.05）;与rs891969位点AA/GA基因型相比,携带GG基因型的个体发生噪声性高频听力损失的风险增加（校正OR=1.650,95%CI=1.032~2.639,P〈0.05）。并按噪声暴露水平、吸烟、饮酒和高血压进行分层分析,未见有统计学意义的结果（P〉0.05）。单体型分析中也未见病例组与对照组单体型频率差异有统计学意义（P〉0.05）。[结论]POU4F3基因单个位点多态性可能不是影响噪声性高频听力损失的遗传易感性因素,但rs1368402和rs891969位点多态性与噪声之间的交互作用可能对其具有重要影响。经Bonferroni检验校正后,上述相关性不再具有统计学意义,研究结果还需进一步验证。
基金supported by the National Natural Science Foundation of China(Nos.81670942,81470704,and 81972057)Special Funds for Taishan Scholar Project.
文摘Objective:The zebrafish is an excellent model for studying gene function in auditory system development.Pou4f3 plays an important role in mouse hair cell formation.Here,we constructed apou4f3-knockoutTg(Brn3c:GFP)zebrafish to provide an efficient fluorescence-visualized model for studying the molecular mechanisms of ear development.Methods:Cas9/single guide RNAs targeting exon 2 ofpou4f3 were designed and injected into one-cell stage zebrafish embryos(G0 generation).The G0 generation were crossed withTg(Brn3c:GFP)zebrafish to obtainpou4f3-mutantTg(Brn3c:GFP)zebrafish.The targeting efficiency was detected by polymerase chain reaction amplification and Sanger sequencing.Zebrafish hair cells were observed by laser scanning confocal microscopyin vivo.The morphology of the otoliths and semicircular canals were analyzed.All animal experiments were approved by the Animal Care and Use Committee of Shandong Provincial Hospital,Cheeloo College of Medicine,Shandong University(approval No.2016-KY-040)on March 3,2016.Results:Thepou4f3-mutantTg(Brn3c:GFP)zebrafish line was successfully established.Fluorescence observation suggested that hair cell development was delayed inpou4f3-knockout zebrafish.Knockout ofpou4f3 also induced defects in the otoliths and semicircular canals and impaired ear function in zebrafish.Conclusion:A CRISPR/Cas9-mediatedpou4f3 mutantTg(Brn3c:GFP)zebrafish model was established for the first time to demonstrate the essential role ofpou4f3 in zebrafish ear development.Our study provides a highly efficient method for the establishment of a visualized model of gene knockout zebrafish and has the potential to allow high-throughput drug screening to explore therapeutics for related diseases.
基金supported by Chinese National Nature Science Foundation(81230020)grant from Minister of Science and Technology of China(2012BAI09B02)to P.D.+2 种基金Chinese National Nature Science Foundation(81371098)Beijing Natural Science Foundation(7132177)Beijing Nova programme(2009B34)to Y.Y.Y
文摘Objective： Based on the clinical manifestations of a hearing loss patient, the POU3F4 gene was tested for diagnosis of etiology. Methods： A comprehensive physical examination was performed on the proband to exclude abnormalities of other organs, and detailed audi- ological testing and temporal bone CT scan were also performed. Genomic DNA was extracted using the proband＇s peripheral blood leukocytes. Polymerase chain reactions （PCR） were performed in the coding sequence of the POU3F4 gene. Direct DNA sequencing was subsequently applied to screen the entire coding region of the POU3F4 gene. Results： The proband had severe sensorineural hearing loss. Temporal CT showed bilateral cochlear incomplete partition, vestibule dysplasia, internal auditory canal fundus expansion, and cochlear interlink with the internal auditory canal fundus. A novel mutation （c.530C 〉 A （p.S 177X）） in the POU3F4 gene was found in this patient, creating an new stop codon and was predicted to result in a truncated protein lacking normal POU3F4 transcription factor function. Conclusion： Through analysis of the POU3F4 gene and clinical manifestations in the patient, we conclude that a novel mutation may have resulted in a premature stop codon, contributing to the mutation of POU3F4 gene.
基金supported by the funding from the National High Technology Research and Development Program of China (863 Program) to Huijun Yuan (No.2007AA02E466)Key Project of National Natural Science Foundation of China to Huijun Yuan (Nos.81030017, 30571018) and Xuezhong Liu (No. 30528025)
文摘We present the clinical and genetic findings for a Chinese family with X-linked non-syndromic hearing loss in which the affected males showed congenital profound sensorineural hearing impairment. In two affected brothers, the computer tomography of temporal bone showed bilateral dilation of the internal auditory canal with fistulous communication between the lateral canal and the basal cochlear turn, which is consistent with the typical DFNX2 phenotype. A missense mutation （c.647G→A） in the POU3F4 gene caused a substitu- tion from glycine to glutamic acid at position 216 （p.G216E）, and this mutation was found to consistently cosegregate with the deafness phenotype in the family. The mutation resulted in the loss of function of the POU3F4 by decreasing the affinity between the protein and DNA, as shown in silico by the structural analysis. Prenatal diagnosis of pregnant proband of this family revealed the c.647G→A mutation in DNA extracted from the amniotic fluid surrounding the fetus. The appropriate use of genetic testing and prenatal diagnosis plays a key role in reducing the recurrence of genetic defects in high-risk families.
基金This work was supported by the grants from the National Key Basic Research Program of China （No.2014CB943001）, the National Natural Science Foundation of China （No. 81120108009 and No. 81530032）, and the China Postdoctoral Science Foundation （No.2015M572690）.
文摘Background： The molecular genetic research showed the association between X-linked bearing loss and mutations in POU3F4. This research aimed to identify a POU3F4 mutation in a nonsyndromic X-linked recessive hearing loss family. Methods： A series of clinical evaluations including medical history, otologic examinations, l：amily history, audiologic testing, and a high-resolution computed tomography scan were performed t＇or each patient. Bidirectional sequencing was carried out for all polymerase chain reaction products or＇the samples. Moreover, 834 controls with normal hearing were also tested. Results： The pedigree showed X-linkage recessive inheritance pattern, and pathogenic mutation （c.499C〉T） was identified in the proband and his family member, which led to a premature termination prior to the entire POU domains. This mutation co-segregated with bearing loss in this family. No mutation ofPOU3F4 gene was found in 834 controls. Conclusions： A nonsense mutation is identified in a family displaying the pedigree consistent with X-linked recessive pattern in POU3F4 gene. In addition, we may provide molecular diagnosis and genetic counseling for this family.
