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OBJECTIVES: A subset of intractable allergic rhinitis (iAR) patients experience severe symptoms which cannot be effectively controlled by standard drug therapy and/or antigen specific immunotherapy. In recent decades, endoscopy vidian neurectomy and posterior nasal nerve neurectomy (PNNN) were introduced as treatments of iAR that have shown to be highly successful at symptom management in a number of patients. But some patients experience relapse or suboptimal symptom control postoperation. To improve the effectiveness of PNNN to control iAR, a modified PNNN surgical approach (mPNNN) combined with accessory posterior nasal nerve neurectomy (aPNNN), which called as mPNNN-aPNNN was used. This study aims to compare the effects between mPNNN-aPNNN and PNNN on controlling the symptoms of iAR and evaluate the surgical effectiveness and safety of mPNNN-aPNNN. METHODS: The patients with iAR experienced mPNNN-aPNNN or PNNN surgery at the department of Otolaryngology Head and Neck Surgery of the Second Xiangya Hospital, Central South University from January 2018 to December 2019 were analyzed retrospectively. The approach of PNNN, a selective resection of the posterior nasal nerve branches, was modified to the neurectomy of total branches of posterior nasal nerve at the sphenopalatine foramen, and combined the operation of aPNNN in which the accessory posterior nasal nerve at the palatine bone perpendicular plate was resect in our study. Daily Nasal Symptom Scores (DNSS), Total Rhinitis Medication Score (TRMS), and the Rhinoconjunctivitis Qualities of Life Questionnaires Scores (RQLQS) were used to evaluate the complications during the operation and after the operation at the 3rd, 6th, 12th, and 24th month postoperatively. Total Nasal Symptom Scores (TNSS) was used to assess the total effective rate and markedly effective rate of the operations. RESULTS: A total of 140 iAR patients experienced mPNNN-aPNNN or PNNN. Those with concomitant septoplasty and/or inferior turbinate reduction, and were absent during the postoperative follow-up were excluded. The final 62 patients with mPNNN-aPNNN and 34 with PNNN were enrolled. DNSS, TNSS, TRMS, and RQLQS at the postoperation were significantly improved compared with the preoperation in all patients (all P<0.001). Compared with PNNN, the postoperative DNSS, TNSS, and TRMS of mPNNN-aPNNN were obviously improved (all P<0.001). There was a persisted relief of symptoms at the postoperation in all patients with mPNNN-aPNNN. The total effective rate and markedly effective rate at the postoperative 24th month were 100% and 83.3%, respectively. Furthermore, the postoperative RQLQS decreased significantly (P<0.001). Only 5 sides of all patients (5/192, 2.6%) reported upper palate numbness during the first week after operation, with all recovered spontaneously in 1 month without treatment. No other postoperative complications occurred in mPNNN-aPNNN and PNNN. CONCLUSIONS: The surgery of mPNNN-aPNNN improve TNSS more significantly than PNNN. The operation of mPNNN-aPNNN is safe and effective to control iAR symptoms.
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Rinite Alérgica , Humanos , Estudos Retrospectivos , Resultado do Tratamento , Rinite Alérgica/cirurgia , Rinite Alérgica/complicações , Conchas Nasais/cirurgia , DenervaçãoRESUMO
OBJECTIVES: To characterize the epithelial-mesenchymal transition (EMT) in chronic rhinosinusitis with nasal polyps (CRSwNP) and to investigate the mechanism by which microRNA-21 (miR-21) regulates EMT in CRSwNP. METHOD: (1) Tissue experiments: Mucosa tissues were collected from 13 patients with CRSwNP and 12 patients with CRS without nasal polyps (CRSsNP), as well as 11 patients without CRS (controls). Protein localization and quantification were achieved by immunofluorescence staining and Western blotting, involving the epithelial marker protein E-cadherin and the mesenchymal marker proteins α-smooth muscle actin (α-SMA), fibronectin, and vimentin. Quantitative RT-PCR was used to detect the relative expression levels of miR-21 and TGF-ß1 mRNAs. (2) Cellular experiments: Primary human nasal epithelial cells (PHNECs) treated with TGF-ß1, or TGF-ß1 with miR-21 inhibitor, or miR-21 mimics alone were observed for morphology changes under a phase-contrast microscope. The expression levels of epithelial/mesenchymal marker proteins were determined as aforementioned. PTEN and phosphorylated Akt were detected by Western blotting. RESULTS: (1) Tissue experiments: Compared with the CRSsNP and control groups, the expression of E-cadherin was downregulated in the CRSwNP group, whereas the expression of TGF-ß1, α-SMA, fibronectin, and vimentin was upregulated. The expression levels of miR-21 and TGF-ß1 mRNAs in CRSwNP were significantly higher than those in CRSsNP and controls. (2) Cellular experiments: TGF-ß1 induced EMT-like transformation in PHNECs, featured by changes in cell morphology and upregulation of mesenchymal proteins and miR-21. The miR-21 inhibitor, as well as the Akt-specific -inhibitor, suppressed TGF-ß1-induced EMT. Mechanically, downregulation of miR-21 resulted in increased PTEN and decreased Akt phosphorylation. Furthermore, overexpression of miR-21 had the opposite effects. CONCLUSIONS: Our findings suggest that the TGF-ß1-miR-21-PTEN-Akt axis may contribute to the pathogenesis of CRSwNP. miR-21 might be a reliable target for treating nasal polyp genesis through EMT suppression. Moreover, miR-21 inhibitors could be a novel class of antipolyp drug that modulates PTEN expression and Akt activation. In addition, further investigation regarding the reason underlying miR-21 overexpression in CRSwNP could provide a molecular target for novel treatment strategies for nasal polyposis.
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Transição Epitelial-Mesenquimal , MicroRNAs/genética , Mucosa Nasal/fisiologia , Pólipos Nasais/imunologia , Rinite/imunologia , Sinusite/imunologia , Fator de Crescimento Transformador beta1/genética , Adolescente , Adulto , Idoso , Células Cultivadas , Criança , Doença Crônica , Feminino , Humanos , Masculino , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Mucosa Nasal/patologia , Pólipos Nasais/genética , PTEN Fosfo-Hidrolase/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Rinite/genética , Sinusite/genética , Fator de Crescimento Transformador beta1/metabolismo , Adulto JovemRESUMO
The TPRN gene encodes taperin, which is prominently present at the taper region of hair cell stereocilia. Mutations in TPRN have been reported to cause autosomal recessive nonsyndromic deafness 79(DFNB 79). To investigate the role of taperin in pathogenesis of hearing loss, we generated TPRN knockout mice using TALEN technique. Sanger sequencing confirmed an 11 bp deletion at nucleotide 177-187 in exon 1 of TPRN, which results in a truncated form of taperin protein. Heterozygous TPRN+/- mice showed apparently normal auditory phenotypes to their wide-type (WT) littermates. Homozygous TPRN-/- mice exhibited progressive sensorineural hearing loss as reflected by auditory brainstem response to both click and tone burst stimuli at postnatal days 15 (P15), 30 (P30), and 60 (P60). Alex Fluor-594 phalloidin labeling showed no obvious difference in hair cell numbers in the cochlea between TPRN-/- mice and WT mice under light microscope. However, scanning electronic microscopy revealed progressive degeneration of inner hair cell stereocilia, from apparently normal at postnatal days 3 (P3) to scattered absence at P15 and further to substantial loss at P30. The outer hair cell stereocilia also showed progressive degeneration, though much less severe, Collectively, we conclude that taperin plays an important role in maintenance of hair cell stereocilia. Establishment of TPRN knockout mice enables further investigation into the function of this gene.
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Surdez/genética , Surdez/patologia , Células Ciliadas Auditivas/ultraestrutura , Perda Auditiva Neurossensorial/genética , Perda Auditiva Neurossensorial/patologia , Proteínas/fisiologia , Estereocílios/patologia , Animais , Células Ciliadas Auditivas/metabolismo , Heterozigoto , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microscopia Eletrônica de Varredura , Proteínas/genética , Deleção de Sequência , Estereocílios/metabolismoRESUMO
Introduction: The pathogenic gene CDH23 plays a pivotal role in tip links, which is indispensable for mechanoelectrical transduction in the hair cells. However, the underlying molecular mechanism and signal regulatory networks that influence deafness is still largely unknown. Methods: In this study, a congenital deafness family, whole exome sequencing revealed a new mutation in the pathogenic gene CDH23, subsequently; the mutation has been validated using Sanger sequencing method. Then CRISPR/Cas9 technology was employed to knockout zebrafish cdh23 gene. Startle response experiment was used to compare with wide-type, the response to sound stimulation between wide-type and cdh23-/-. To further illustrate the molecular mechanisms underlying congenital deafness, comparative transcriptomic profiling and multiple bioinformatics analyses were performed. Results: The YO-PRO-1 assay result showed that in cdh23 deficient embryos, the YO-PRO-1 signal in inner ear and lateral line neuromast hair cells were completely lost. Startle response experiment showed that compared with wide-type, the response to sound stimulation decreased significantly in cdh23 mutant larvae. Comparative transcriptomic showed that the candidate genes such as atp1b2b and myof could affect hearing by regulating ATP production and purine metabolism in a synergetic way with cdh23. RT-qPCR results further confirmed the transcriptomics results. Further compensatory experiment showed that ATP treated cdh23-/- embryos can partially recover the mutant phenotype. Conclusion: In conclusion, our study may shed light on deciphering the principal mechanism and provide a potential therapeutic method for congenital hearing loss under the condition of CDH23 mutation.
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Background: Radioresistance in head and neck squamous cell carcinoma (HNSCC) patients means response failure to current treatment. In order to screen radioresistant biomarkers and mechanisms associated with HNSCC, differentially expressed genes (DEGs) associated with radioresistance in HNSCC were investigated. Methods: The HNSCC cell line with radioresistance, Hep2-R, was established and detected the radiosensitivity using MTT, colony formation assay and flow cytometry analysis. Clariom™ D chip was applied to compare DEGs between Hep2 and Hep2-R groups and build the differential gene expression profiles associated with radioresistance in HNSCC. Bioinformatic analysis were used to find biological functions and pathways that related to radioresistance in HNSCC, including cell adhesion, cytochrome P450 and drug metabolism. Gene Expression Omnibus (GEO) datasets were selected to verify DEGs between HNSCC radioresistant cells and tissues. The representation of DEGs were validated between HNSCC patients with complete response and post-operative radiation therapy failure. In addition, we evaluated the clinical prognosis of DEGs using The Cancer Genome Atlas (TCGA) database. Results: 2,360 DEGs (|Fold Change|>1.5, p < 0.05) were identified between Hep2 and Hep2-R, including 1,144 upregulated DEGs and 1,216 downregulated DEGs. They were further verified by HNSCC radioresistant cells and tissues in GEO. 13 radioresistant DEGs showed same difference in expression level between cells and tissues. By comparing 13 DEGs with HNSCC patients, upregulations of FN1, SOX4 and ETV5 were found identical with above results. Only FN1 was a prognostic indicator of HNSCC in TCGA. Conclusion: FN1 is the potential novel biomarker for predicting poor prognosis and radioresistance in HNSCC patients. Overexpression of FN1 plays an important role in the tumorigenesis, prognosis and radioresistance of HNSCC.
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OBJECTIVE: To establish the kanamycin-induced deafness model in SD rats, and to investigate the expression and significance of transmembrane protease, serine 3 (TMPRSS3) in the cochlea following kanamycin ototoxicity. METHODS: A total of 40 male SD rats were randomly divided into 4 groups. The experimental rats received intramuscular kanamycin sulfate for 3, 7, and 14 consecutive days, and the control group were treated with normal saline for 14 days. Auditory brainstem responses (ABR) were obtained before and after the kanamycin administration. The expression of TMPRSS3 in the cochlea was identified and detected by immunohistochemistry and Western blot. RESULTS: Kanamycin-induced deafness model in the SD rats was successfully established. ABR thresholds were increased and the expression of TMPRSS3 in the cochlea was reduced after the kanamycin injection (P<0.01). CONCLUSION: TMPRSS3 may play an important role in normal cochlea function and involve in the process of aminoglycoside antibiotics induced deafness.
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Antibacterianos/toxicidade , Cóclea/metabolismo , Surdez/metabolismo , Canamicina/toxicidade , Proteínas de Membrana/metabolismo , Serina Endopeptidases/metabolismo , Animais , Cóclea/efeitos dos fármacos , Surdez/induzido quimicamente , Modelos Animais de Doenças , Potenciais Evocados Auditivos do Tronco Encefálico/fisiologia , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: To report a novel deafness-causing mutation c.465T>A, p.Y155X in connexin 26 (CX26) (also called gap junction protein beta-2, GJB2), and perform functional analysis of the mutated protein p.Y155X in Hela cells to explore the underlying mechanism on deafness. METHODS: Mutations in CX26 gene of the proband in an autosomal recessive inherited deafness family were tested by direct DNA sequencing method. Mutant p.Y155X, which was found in the deafness family, and wild type CX26 (wtCX26), were directionally subcloned into the pEGFP-N1 plasmid to construct the recombinant fusion protein expression vector of CX26 p.Y155X-EGFP and wtCX26-EGFP, followed by transfecting into HeLa cells. The expression of the mutated and wild type proteins was analyzed using Western blot analysis. The intracellular localization of proteins and the formation of gap junction-like plaques at plasma membrane were observed under confocal microscope. Gap junction coupling was tested by calcein-AM dye transfer experiment. RESULTS: A novel nonsense mutation c.465T>A, p.Y155X in the CX26 gene was found in the autosomal recessive deafness family. The molecular weight of protein p.Y155X was smaller than that of wtCX26 in transiently expressed HeLa cells. The mutated protein failed to reach the cell surface to form gap junction plaques, and displayed cytoplasmic accumulation. Also, no calcein-AM dye was transferred from the donor cells to the recipient cells when both were transfected with CX26 p.Y155X. The wtCX26 protein localized at the cell membrane to form gap junction plaques with permeability to fluorescent dye calcein AM. CONCLUSION: CX26 p.Y155X could not be targeted to the plasma membrane and there was no formation of gap junction channels between the adjacent cells. The mutation c.465T>A, p.Y155X in CX26 gene was responsible for the autosomal recessive hearing impairment in this family.
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Conexinas/genética , Surdez/genética , Sequência de Aminoácidos , Criança , Códon sem Sentido/genética , Conexina 26 , Análise Mutacional de DNA , Feminino , Células HeLa , Humanos , Masculino , Dados de Sequência Molecular , Linhagem , Homologia de Sequência de AminoácidosRESUMO
OBJECTIVE: To identify the genetic characteristics in patients with nonsydromic hearing loss (NSHL) in Hunan province, to determine the prevalence and spectrum of mutations in GJB2 gene, and to explore the pathogenic mechanism. METHODS: A total of 140 sporadic patients with NSHL were enrolled after clinical examination. Molecular studies were performed by amplifing the coding region of GJB2 gene, purifying the PCR products, and sequencing directly. Sequences were analysed by DNAStar software to determine GJB2 mutations in the patients. Special method was designed to confirm the unreported mutation. RESULTS: We detected GJB2 mutation in 56 out of the 140 patients (40%, 56/140). Both of the 2 alleles were mutated in 29 patients and 1 allele in the other 27 patients, and the rate of allele mutation was 30.4%(85/280). Ten variations were detected, including 7 mutations and 3 polymorphisms. The deaf-causing mutations were nonsense mutation c.139G>T; frameshift mutation c.235delC and c.176-191del16; and missense mutation c.109G>A, c.344T>G, c.550C>T and c.571T>C. The unreported missense mutation was c.344T>G. The c.235delC mutation was the most prevalent mutation found in the 27 patients (19.3%, 27/140). The frequency of c.109G>A mutation was next to c.235delC found in 25 patients (17.9%, 25/140). CONCLUSION: GJB2 mutation is a major cause for NSHL. The most common-spot in Chinese patients with NSHL is c.235delC. The unreported missense mutation is c.344T>G.
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Conexinas/genética , Perda Auditiva/genética , Mutação Puntual/genética , Sequência de Bases , China , Conexina 26 , Análise Mutacional de DNA , Deleção de Genes , Humanos , Dados de Sequência Molecular , Mutação de Sentido IncorretoRESUMO
CONCLUSION: The results show that alpha1D, alpha1E, alpha2/delta, beta1, and beta3 subunits are expressed in spiral ganglion cells (SGCs), and the coexpression of alpha1D and alpha1E suggests the presence of L-type and R-type calcium channels in mammalian SGCs. OBJECTIVE: To investigate the types of subunits of voltage-gated calcium channels in SGCs of the mouse. MATERIALS AND METHODS: SGCs were isolated from cochleae of neonatal mice and cultured for 24 h. Total RNA was extracted from cultured cells. After reverse transcription, the resulting cDNA was amplified by PCR with primers targeted to nucleotide sequences corresponding to seven different calcium channel subunits. The types of calcium channel subunits were identified by PCR analysis and nucleotide sequencing. RESULTS: RT-PCR showed the strong and consistent amplification of alpha1D, alpha1E, alpha2/delta, beta1, and beta3 subunits from the mRNA of SGCs, and nucleotide sequencing confirmed the identity of mouse cochlear subunit cDNAs.
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Canais de Cálcio Tipo R/genética , Canais de Cálcio Tipo R/metabolismo , Canais de Cálcio/genética , Canais de Cálcio/metabolismo , Proteínas de Transporte de Cátions/genética , Proteínas de Transporte de Cátions/metabolismo , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , Gânglio Espiral da Cóclea/citologia , Gânglio Espiral da Cóclea/metabolismo , Animais , Animais Recém-Nascidos , Canais de Cálcio Tipo L/genética , Canais de Cálcio Tipo L/metabolismo , Cóclea/citologia , Cóclea/metabolismo , DNA Complementar/genética , DNA Complementar/metabolismo , Camundongos , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodosRESUMO
OBJECTIVE: To review the surgical treatment for reconstructing hypopharynx and cervical esophagus after hypopharyngo-oesophagectomy, and to evalue its efficacy. METHODS: Different methods were adopted to reconstruct the hypopharynx and cervical esophagus among 25 cases, including 14 cases of carcinoma of the hypopharynx and 11 of carcinoma of hypopharynx and cervical esophagus. In accordance with the standard of the International Union Against Cancer in 1997, the 25 cases were divided into different clinic stages, among which 5 were in T(2)N(0), 2 in T(2)N(1), 4 in T(3)N(0), 3 in T(3)N(1), 7 in T(4)N(1) and 3 in T(4)N(2). Treatment protocol was as follow: Pure operation for 5 cases, re-operation after radiotherapy for 2 cases, operation plus radiotherapy for 18 cases, laryngeal conservation operation for 8, and neck dissection for 21 cases. Reconstruction was done by using free jejunal transplantation, gastric pull-up, the laryngotracheal flap, and myocutaneous flap. RESULTS: After the reconstruction, 3 cases of free jejunal graft and gastric pull-up, 4 of laryngotracheal flap recovered oral fleeding within 2 weeks. No serious complications occurred. After 18 cases underwent the myocutaneous flap reconstruction, no complications occurred in 10 patients, but there were different complications in 8 cases, including pharyngocutaneous fistula (6 cases), haryngoesphageal stenosis (7 cases), and pectoralis major myocutaneous flap necrotic (1 case). The 3-year survival rate was 38.9% (7/18). CONCLUSION: Reconstruction with free jejunal graft, gastric pull-up, and laryngotracheal flap constitutes is a safe and reliable method to restore the continuity of the upper digestive tract after pharyngo-laryngo-oesophagectomy. After the reconstruction with myocutaneous flap, there is high incidence of pharyngocutaneous fistula and haryngoesophageal stenosis.
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Neoplasias Esofágicas/cirurgia , Esofagoplastia/métodos , Neoplasias Hipofaríngeas/cirurgia , Hipofaringe/cirurgia , Adulto , Idoso , Carcinoma de Células Escamosas/cirurgia , Esôfago/cirurgia , Feminino , Humanos , Jejuno/transplante , Masculino , Pessoa de Meia-Idade , Procedimentos de Cirurgia Plástica/métodos , Retalhos CirúrgicosRESUMO
OBJECTIVE: To screen and identify the proteins that interact with connexin 26 (CX26) and to analyze the expressions of these proteins in cochlea so as to explore the proteins that relate to the trafficking, assembly, localizing and gap junction functions of CX26. METHODS: The whole coding region of GJB2 (CX26) gene was amplified from normal human genomic DNA by polymerase chain reaction (PCR) and then directionally subcloned into the vector pGBKT7 plasmid of the Match Maker Ga14 Two-Hybrid System 3 as a target to screen the interactive proteins of CX26 from the human fetal brain cDNA library by the yeast two hybrid technique. The false positive clones were discarded from the preys by repeated yeast two hybrid method between CX26 and everyone of the preys respectively. The DNAs of the insert of the identified positive clone were sequenced and BLAST analyzed against the GenBank. Lastly, the mRNA of the gene encoding the identified protein was analyzed in the murine inner ear by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The insert of one positive clone contained 867 bp with the former 525 bp being coding region. The DNA sequence and the open reading frame of the insert were identical to the 525 bp before the stop codes (including the stop codes) and the 238 bp after the stop codes of RTN4 gene which encoded Nogo protein. And the 174 amino acid residues encoded by the insert were those of the C-terminal of Nogo protein: Nogo-A, Nogo-B and Nogo-C. RTN4 mRNA expressed in the murine inner ear was confirmed by RT-PCR method. CONCLUSION: The C-terminal of Nogo protein interacts with CX26. Nogo protein expresses in the inner ear and may take part in the trafficking of CX26 or CX26 gap junction function.
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Conexinas/metabolismo , Orelha Interna/metabolismo , Proteínas da Mielina/metabolismo , RNA Mensageiro/genética , Animais , Sequência de Bases , Conexina 26 , Conexinas/genética , Expressão Gênica , Humanos , Camundongos , Dados de Sequência Molecular , Proteínas da Mielina/genética , Proteínas Nogo , Ligação Proteica , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Técnicas do Sistema de Duplo-HíbridoRESUMO
OBJECTIVE: The present study aims to test whether deaf children with unilateral cochlear implantation (CI) have higher intelligence quotients (IQ). We also try to find out the predictive factors of intelligence development in deaf children with CI. METHODS: Totally, 186 children were enrolled into this study. They were divided into 3 groups: CI group (N = 66), hearing loss group (N = 54) and normal hearing group (N = 66). All children took the Hiskey-Nebraska Test of Learning Aptitude to assess the IQ. After that, we used Deafness gene chip, Categories of Auditory Performance (CAP) and Speech Intelligibility Rating (SIR) methods to evaluate the genotype, auditory and speech performance, respectively. RESULTS: At baseline, the average IQ of hearing loss group (HL), CI group, normal hearing (NH) group were 98.3 ± 9.23, 100.03 ± 12.13 and 109.89 ± 10.56, while NH group scored higher significantly than HL and CI groups (p < 0.05). After 12 months, the average IQ of HL group, CI group, NH group were99.54 ± 9.38,111.85 ± 15.38, and 112.08 ± 8.51, respectively. No significant difference between the IQ of the CI and NH groups was found (p > 0.05). The growth of SIR was positive correlated with the growth of IQ (r = 0.247, p = 0.046), while no significant correlation were found between IQ growth and other possible factors, i.e. gender, age of CI, use of hearing aid, genotype, implant device type, inner ear malformation and CAP growth (p > 0.05). CONCLUSIONS: Our study suggests that CI potentially improves the intelligence development in deaf children. Speech performance growth is significantly correlated with IQ growth of CI children. Deaf children accepted CI before 6 years can achieve a satisfying and undifferentiated short-term (12 months) development of intelligence.
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Desenvolvimento Infantil , Implante Coclear , Surdez/cirurgia , Auxiliares de Audição , Perda Auditiva/reabilitação , Inteligência , Fatores Etários , Estudos de Casos e Controles , Criança , Pré-Escolar , Implantes Cocleares , Surdez/psicologia , Feminino , Perda Auditiva/psicologia , Humanos , Testes de Inteligência , Masculino , Inteligibilidade da Fala , Percepção da FalaRESUMO
OBJECTIVE: To explore the value of a combined computed tomography (CT) and magnetic resonance imaging (MRI) in evaluating profound sensorineural deafness patients before cochlear implant (CI) surgery. METHODS: A retrospective analysis of 1012 cases of profound sensorineural deafness that received CI was performed. RESULTS: A total of 96 cases were diagnosed with inner ear abnormalities including large vestibular aqueduct syndrome (LVAS, n = 61), Michel deformity (n = 3), cochlear incomplete partition I (n = 2), cochlear incomplete partition II (n = 6), cochlear hypoplasia with vestibular malformation (n = 3), cochlear ossification (n = 3), bilateral internal auditory canal obstruction (n = 5) and internal auditory canal stenosis (n = 2). CONCLUSION: High resolution CT (HRCT) can display bony structures while MRI can image the membranous labyrinth in preoperative evaluation for cochlear implantation. The combination of these two modalities provides reliable anatomical information regarding the bony and membranous labyrinths, as well as the auditory nerve.
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OBJECTIVES: To review the identified deafness genes related to nonsyndromic hearing loss (NSHL) and summarize their expressions and functions in the cochlea and to introduce the current studies of molecular genetics on NSHL in China. METHODS: The presented data are based on a review of the literature as well as the author' s experience with NSHL and communications with other researchers in China over the past 3 years. RESULTS: Currently, 23 deafness genes related to NSHL have been cloned and identified. Some genes are associated with both NSHL and syndromic hearing loss (SHL), in both dominant and recessive deafness. Deafness genes have a highly specific expression pattern in the inner ear. Some functional categories are starting to emerge from a characterization of deafness genes. There are interacting genes in the genetic background that influence the extent of hearing impairment. The GJB3 gene, which is associated with high-frequency hearing impairment, was cloned in a Chinese laboratory. Mutations in some genes, such as GJB2 and mitochondrial 12S rRNA, have been screened in Chinese patients with NSHL. Mapping new deafness gene loci as well as identifying new genes and their functions is an active area of study in China. CONCLUSIONS: It is challenging for us to continue identifying new deafness genes and analyze gene functions. By identifying genes responsible for monogenic hearing impairment, more insight may be gained into the molecular process of hearing and the pathology of hearing loss.
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Surdez/genética , Mapeamento Cromossômico , Clonagem Molecular , Conexina 26 , Conexina 30 , Conexinas/genética , Humanos , MutaçãoRESUMO
BACKGROUND: Mutations in GJB2 gene are a major cause of autosomal recessive congenital hearing loss and the cause in some rare cases of the autosomal dominant form. The purpose of this study was to investigate the frequency and the features of GJB2 mutations in the Chinese patients with congenital sensorineural deafness. METHODS: Using PCR amplifying the entire coding region of GJB2 gene and direct DNA sequencing to analyze mutations in this gene among unrelated 69 cases with autosomal recessive congenital nonsyndromic deafness and 27 cases of dominant congenital deafness and 35 sporadic cases. We also detected mutations in GJB2 in 100 control subjects with normal hearing. RESULTS: 17.4% (12/69) of the probands in the autosomal recessive, 7.4% (2/27) of dominant families and 5.7% (2/35) of the sporadic congenital deafness patients had deafness-causing mutations in GJB2, respectively. Nine types of the mutations in GJB2 were detected in the recessive and sporadic group. They consisted of five types of polymorphism, and four types of deafness-causing mutation with homozygous 35delG in 1 sporadic (1/35), and 235delC frameshift mutation in 1 sporadic (homozygotes) and 10 recessive patients (2 heterozygotes and 8 homozygotes), and homozygous 442G-->A missense mutation and homozygous 465T-->A nonsense mutation in 1 different recessive proband, respectively. The 465T-->A that related to recessive deafness was a novel mutation found by this study. The homozygous (10/69, 14.5%) and the heterozygous (2/69, 2.9%) GJB2 mutation in the recessive patients (12/69, 17.4%) and the homozygotes in the sporadic patient (2/35, 5.7%) all had congenital severe to profound sensorineural hearing loss. 511G-->A missense mutation and 299-300delAT frameshift mutation were found in two autosomal dominant congenital deafness families (2/27, 7.4%). The total mutation frequency of GJB2 was 12.2% (16/131) in the Chinese patients with congenital sensorineural deafness and 235delC was the most common deafness-causing mutation. Six types of mutation-5 types of polymorphism and 1 type of heterozygous deletion (235delC) mutation were found in the 100 control subjects. The carry rate of the most frequent type of mutation 235delC was 0.5% in the controls (1/200 alleles). 109G-->A was the most frequent (15/100, 15%) and 79G-->A was the second common (8/100, 8%) polymorphism in this population. CONCLUSIONS: The general mutation rate of GJB2 is 12.2% (16/131) and the 235delC is the most common type of deafness-causing mutation in Chinese patients with congenital hearing loss. 465T-->A nonsense mutation that is associated to autosomal recessive deafness is a novel mutation found by this screening. 511G-->A and 299-300delAT mutations contribute to autosomal dominant hearing loss. The study further supports the view that the common types of mutation in GJB2 according to different ethnic background and that the mutation prevalence in the East Asian deafness population is lower than that in the white population.
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Conexinas/genética , Perda Auditiva Neurossensorial/genética , Mutação , Conexina 26 , HumanosRESUMO
OBJECTIVE: To screen and identify the interactive proteins with connexin 26 (Cx26) by the yeast two hybrid technique. METHODS: The whole coding region of Cx26 (GJB2) gene was amplified from normal human genomic DNA by polymerase chain reaction (PCR). The "bait" Cx26 was then subcloned into the vector pGBKT7 plasmid of the MatchMaker Gal4 Two-Hybrid System 3 as a target to screen its interactive proteins ("prey") from the human fetal brain eDNA library by the yeast two hybrid technique. The false positive clones were discarded from the preys by one to one yeast two hybrid method between Cx26 and the preys. The DNAs of the preys were sequenced and BLAST analyzed against the GenBank, and also underwent other bioinformatics analysis. RESULTS: The insert of one positive clone contained 145 amino acids residues that was identical to the C-terminal of the neuroendocrine specific protein (NSP) and the open reading frame of the insert was correct. CONCLUSION: Cx26 is interacted with the C-terminal of NSP. NSP may participate in the process of Cx26 transportation, assembling the connexon, or influencing the functions of its connexons.
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Encéfalo/metabolismo , Conexinas/química , Proteínas do Tecido Nervoso/química , Mapeamento de Interação de Proteínas , Oxirredutases do Álcool , Sequência de Aminoácidos , Sequência de Bases , Conexina 26 , Conexinas/genética , Proteínas de Ligação a DNA/química , Feto , Biblioteca Gênica , Humanos , Dados de Sequência Molecular , Proteínas do Tecido Nervoso/genética , Fosfoproteínas/química , Ligação Proteica , Técnicas do Sistema de Duplo-Híbrido , Leveduras/genéticaRESUMO
OBJECTIVE: To review the clinical manifestations and management of nasal sinus mucoceles with visual loss. METHOD: Medical records for 23 patients of paranasal sinus mucoceles with visual impairment were re viewed retrospectively during 8-year period (from 2002 to 2010). Ten mucoceles were found in the frontal or fronto-ethmoidal sinuses, 6 in the ethmoidal sinuses, 7 in the sphenoidal or spheno-ethmoidal sinuses. Because the majority of early chief complaints were problems related to vision, patients were often seen by ophthalmologists first. Poor vision was more common in patients with sphenoid or spheno-ethmoidal sinus mucoceles because of their proximity to the optic nerve. CT and MRI were important tools for diagnosing nasal sinus mucocele. The patients received endoscopic surgery to remove mucocele and to decompress the optic nerve. Steroid therapy was given postoperatively and routine examination with endoscopy were carried out during follow-up. RESULT: Postoperatively, the majority of symptoms, such as exophthalmos, epiphora, diplopia and headache, disappeared in all patients. However, vision recovery was observed only in some patients. Recovery of vision depended on the timing of surgery and severity of initial visual loss. Delay in treatment can seriously compromise recovery of vision impairment. Moreover, patients without light perception before surgery had poor visual recovery even if optic nerve decompressions were performed. CONCLUSION: Endoscopic surgery is effective to nasal sinus mucocele with visual loss. Because visual recovery depends on prompt diagnosis and surgical intervention, a good understanding of the disease and prompt imaging studies are important.
Assuntos
Cistos/cirurgia , Doenças dos Seios Paranasais/diagnóstico , Doenças dos Seios Paranasais/cirurgia , Adolescente , Adulto , Cistos/complicações , Cistos/diagnóstico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Doenças dos Seios Paranasais/complicações , Estudos Retrospectivos , Baixa Visão/etiologia , Adulto JovemRESUMO
OBJECTIVE: To investigate the expressions of stathmin gene and its coding protein in laryngeal squamous cell carcinoma, and to explore the relationship between stathmin gene and the biological behaviors of laryngeal squamous cell carcinoma for understanding the tumorigenicity and development of laryngeal squamous cell carcinoma. METHODS: Laryngeal carcinoma tissues (studying group) in the tumors center and laryngeal normal tissues (control group) parted from 1.0 cm of the safe borderline of the tumors were took from 38 patients with laryngeal squamous cell carcinoma while they were in operation. Semiquantitative method of reverse transcriptase polymerase chain reaction (RT-PCR) was used to analyze the expression level of stathmin mRNA, and immunohistochemical staining (frozen section) was used to detect the expressions of stathmin protein, in laryngeal carcinoma tissues and laryngeal normal tissues of 38 cases, respectively. RESULTS: mRNA of stathmin gene was all positively expressed in laryngeal carcinoma tissues and in laryngeal normal tissues of 38 cases by RT-PCR. However, stathmin mRNA was obviously overexpressed in laryngeal carcinoma tissues than that in laryngeal normal tissues (t = 9.655, P < 0.05). Immunohistochemical staining showed stathmin protein was positively expressed in laryngeal carcinoma tissues of 26 cases (26/38, 68.4%), and mild-positively expressed in laryngeal normal tissues in 13 cases (13/38, 34.2%). There was significant difference between the expression rate of stathmin protein in laryngeal carcinoma tissues and in laryngeal normal tissues (chi2 = 8.901, P < 0.05). Meanwhile, the expression level of stathmin mRNA and the positive-expressed rate of stathmin protein in laryngeal carcinoma tissues of the advanced stage patients group (III stage and IV stage) were significantly higher than these in laryngeal carcinoma tissues of I and II stage patients group (t = 6.284, chi2 = 5.810, P < 0.05), and they were also significantly higher in laryngeal carcinoma tissues of the patients group with cervical lymph node metastasis than in laryngeal carcinoma tissues of the patients group without cervical lymph node metastasis (t = 9.350, chi2 = 6.923, P < 0.05). CONCLUSIONS: The expression levels of stathmin gene and protein were significantly higher in laryngeal squamous cell carcinoma than these in laryngeal normal tissues, the levels are also significantly higher in advanced stage patients group (III stage and IV stage) than in the early stage patients group (I and II), and they are also related to the cervical lymph node metastasis of carcinoma. Stathmin gene may play an important role in the pathogenesis and development of laryngeal carcinoma and may be related to its prognosis.
Assuntos
Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Neoplasias Laríngeas/genética , Neoplasias Laríngeas/metabolismo , Estatmina/metabolismo , Adulto , Idoso , Carcinoma de Células Escamosas/patologia , Humanos , Neoplasias Laríngeas/patologia , Pessoa de Meia-Idade , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Estatmina/genéticaRESUMO
OBJECTIVE: To enhance the cure rate and lower the complication rate and the mortality rate through summarizing the clinical features and experiences in diagnosis and therapy of carotid body tumor (CBT). METHOD: Retrospectively analyzed the clinical data of 21 cases (23 sides) of CBT from 1995-2095 occurring in our hospital. RESULT: The accurate diagnosis rates hy using digital subtraction angiography (DSA) and magnetic resonance imaging (MRI) were 100%. Seventeen cases (19 sides) accepted surgical operation with different kinds of procedures. The tumors of 8 cases were simplex isolated from the carotid artery. Both the tumour and the external carotid artery were resected in 9 cases. One case underwent resection of both the internal and external carotid artery and the tumour without carotid reconstruction. One case underwent resection of the internal, external carotid artery and the tumor with reconstruction of the internal carotid artery. No operative mortality was observed. The ventricular arrhythmia which had not been controlled pre-operation occurred in 1 case who was finally self-cured. One case had hoarseness and completely recovered in one week. and 1 case without carotid reconstruction had a frequent headache and gradually recovered in 5 months. The others had no complications. CONCLUSION: OSA and MRI are the best methods for diagnosing CBT. Surgery is the first choice concerning the treatment of CBT. Accurate preoperative evaluation, correct therapeutic decision exquisite vascular surgical techniques can help to significantly decrease, even avoid the complications.
Assuntos
Tumor do Corpo Carotídeo/diagnóstico , Tumor do Corpo Carotídeo/cirurgia , Adolescente , Adulto , Idoso , Angiografia Digital , Tumor do Corpo Carotídeo/diagnóstico por imagem , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Adulto JovemRESUMO
OBJECTIVE: To investigate subunit type of voltage-dependent calcium channels in the spiral ganglion cells of the mouse. METHODS: The spiral ganglion cells were dissected from cochleae of neonatal mice and cultured for 24 h. Total RNA was extracted from cultured spiral ganglion cells. After reverse transcription, resulting cDNA was amplified by polymerase chain reaction (PCR) with primers targeted to nucleotide sequences corresponding to 7 different calcium channel subunits. The types of calcium channel subunits were identified by PCR analysis and nucleotide sequencing. RESULTS: Reverse transcription (RT)-PCR products representing subunit gene expression were strongly and consistently amplified for alpha1 D, alpha1 E, alpha2/delta, beta1 and beta3. Nucleotide sequencing confirmed the identity of mouse cochlear subunit cDNAs. CONCLUSIONS: alpha1D, alpha1E, alpha2/delta, beta1 and beta3 subunits are expressed in spiral ganglion cells. And the coexpression of alpha1D and alpha1 E demonstrate the presence of L-type and R-type calcium channels in mammalian spiral ganglion cells.