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1.
Acta Otolaryngol ; 135(5): 451-8, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25739867

RESUMO

CONCLUSION: The hearing conditions of the centenarians were quite poor as regards hearing thresholds and speech detection ability. OBJECTIVE: To investigate hearing conditions of centenarians. METHODS: A total of 54 centenarians in Rizhao and Linyi Districts in Shandong Province were investigated to assess hearing conditions of centenerians comprehensively by questionnaire investigation, pure-tone audiometry, acoustic immitance, intelligence evaluation, and speech detection scores. Also, 135 individuals were recruited as controls and divided into four groups according to their age: 45-59 years, 60-69 years, 70-79 years, and 80-89 years. RESULTS: The hearing thresholds of the centenarians were dramatically higher than those of the control group (p < 0.05) and all centenarians suffered moderate to profound hearing loss according to the World Health Organization (WHO) criteria. Few centenarians had normal level of speech detection scores. All centenarians showed descending hearing curve, and the hearing threshold of the male centenarians at 8000 Hz was higher than that of the females (p = 0.047). There was a significant air-bone conduction gap in the centenarians (p < 0.05).


Assuntos
Limiar Auditivo , Testes Auditivos , Presbiacusia/diagnóstico , Presbiacusia/epidemiologia , Testes de Impedância Acústica , Idoso , Idoso de 80 Anos ou mais , Audiometria de Tons Puros , Condução Óssea , Estudos de Casos e Controles , China , Estudos Transversais , Feminino , Humanos , Inteligência , Masculino , Pessoa de Meia-Idade , Valores de Referência , Reflexo Acústico , Teste do Limiar de Recepção da Fala
2.
Cell Biochem Biophys ; 70(3): 1813-5, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25269771

RESUMO

To investigate the pharmacodynamic effects of Binghuang ear drop on acute suppurative otitis externa in guinea pig model. Thirty guinea pigs were randomly divided into three groups, with ten animals in each group. Group A animals had normal ear canal and Binghuang ear drops (two drops, B.I.D) were applied in both ears for 7 days; Group B animals had induced otitis externa and received identical prescription as group A; Group C had normal ear canal and were treated with normal saline (two drops, B.I.D) for 7 days. After the treatments, the external morphology of ear canals was observed and the paraffin sections of external auditory canal were prepared and examined under the microscope. The inflammatory manifestation and cell infiltration into the skin of group B was significantly attenuated after the Binghuang ear drops treatment. In contrast, no allergy or side effects were produced by Binghuang ear drops application in the animals with normal ear canals. Binghuang ear drops could be used to treat acute otitis externa by eliciting anti-bacterial effects.


Assuntos
Antibacterianos/farmacocinética , Otite Externa/patologia , Doença Aguda , Animais , Modelos Animais de Doenças , Meato Acústico Externo/efeitos dos fármacos , Meato Acústico Externo/patologia , Cobaias , Meia-Vida , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/patogenicidade , Otite Externa/etiologia , Cloreto de Sódio/farmacologia
3.
Acta Otolaryngol ; 131(11): 1136-41, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21838606

RESUMO

CONCLUSION: Bone marrow mesenchymal stem cells (MSCs) have the ability to differentiate into hair cells, and this method of culturing MSCs provides a useful tool for studies on mammalian cochlear hair cell regeneration. OBJECTIVE: To investigate a method to induce bone marrow MSCs to differentiate into inner ear hair cells. METHODS: Rat bone marrow MSCs were isolated from healthy rats and cultured in vitro. To make sure that the cultured cells were bone marrow MSCs, the expression of MSC markers such as SH2, CD31, CD34, and CD44 genes on the cultured cells was assessed by RT-PCR. Adipogenic cells and osteogenic cells were induced by the differentiation of the cultured cells, respectively, suggesting that the cultured cells have the characteristic of pluripotent differentiation. Then they were induced to differentiate into neural stem cells and hair cell progenitor cells. Immunohistochemistry experiments were carried out to detect the expression of molecular markers. Scanning electron microscope samples were prepared for observation of the morphology of the cells. RESULTS: Rat bone marrow MSCs were successfully isolated, purified, cultured, and identified in vitro. They were also successfully induced to differentiate into neural progenitor cells and then hair cell-like cells that expressed myosin VIIa.


Assuntos
Diferenciação Celular , Células Ciliadas Auditivas Internas/citologia , Células-Tronco Mesenquimais/fisiologia , Animais , Células da Medula Óssea/fisiologia , Células Cultivadas , Imuno-Histoquímica , Células-Tronco Mesenquimais/citologia , Ratos
4.
Int J Dev Neurosci ; 29(1): 15-23, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20969946

RESUMO

The regulation of the bone morphogenetic protein (BMP) signal transduction pathway is important in the development of the inner ear and vestibular system. We reported previously that small mothers against decapentaplegic homolog-4 (Smad4) is required for inner ear cochlear development and normal auditory function in mammals; however, the distribution and functional mechanisms of Smad4 at various stages of vestibular development remained unclear. To investigate the relationship between the Smad4 gene and vestibular organ development, we measured changes in the expression of BMP4 and Smad4 during vestibular development in C57BL/6 mice. In addition, vestibular structures, pathologic changes, and the vestibular function of chondrocyte-specific Smad4 knockout mice were compared to those of the control group. We found that the expression of Smad4 in the inner ear was delayed compared with that of BMP4. Moreover, chondrocyte-specific Smad4 knockout homozygous mice showed stunted growth and partial vestibular deformities, but it showed less histologic changes in the vestibular end-organs and saccule dysfunction. These results suggest that Smad4 participates in late-stage shaping of the configuration of the vestibule and development of vestibular functional, but a Smad4-independent pathway for the inner ear vestibular BMP4 signal transduction could not be rule out.


Assuntos
Proteína Smad4/metabolismo , Vestíbulo do Labirinto/anatomia & histologia , Vestíbulo do Labirinto/embriologia , Vestíbulo do Labirinto/fisiologia , Animais , Comportamento Animal/fisiologia , Proteína Morfogenética Óssea 4/genética , Proteína Morfogenética Óssea 4/metabolismo , Potenciais Evocados/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Transdução de Sinais/fisiologia , Proteína Smad4/genética
5.
Artigo em Chinês | MEDLINE | ID: mdl-21176571

RESUMO

OBJECTIVE: To explore the safety and efficacy of cochlear implantation among elderly patients with severe to profound hearing loss. METHODS: Eight pre-elderly and elderly patients with an medium age of 58 years who suffered from bilateral severe to profound sensorineural hearing loss received cochlear implantation between November 2008 and November 2009. The patients' tolerance to implant surgery and the occurrence of complications were observed. Three months after switch-on, aided threshold and speech performance were measured. RESULTS: The surgery was uneventful in all cases with normal intraoperative neural response telemetry elicited. Three months after switch-on, average aided threshold across speech frequencies was 35 - 50 dB HL measured in sound field with warble tone. The results of speech audiometry showed large variation between individuals. Some patients achieved good performance in monosyllable recognition test, disyllables threshold test and sentences recognition test under both bubble noise and quiet conditions. CONCLUSIONS: Pre-elderly and elderly patients can endure a state of general anesthesia for cochlear surgery without complications. Cochlear implant can provide reconstruction of speech recognition capabilities for elderly patients suffering from severe to profound hearing loss. Cochlear implantation can improve the quality of life of elderly patients with hearing loss.


Assuntos
Implante Coclear , Perda Auditiva/reabilitação , Idoso , Implantes Cocleares , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento
6.
J Nanosci Nanotechnol ; 10(11): 7262-5, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21137911

RESUMO

Mammalian cochlear hair cells don't regenerate naturally after injury, which usually leave permanent hearing loss. Math1 gene is a positive regulator of hair cell differentiation during cochlear development and was proved to be very critical in hair cell regeneration in deaf animals. Generating new cochlear hair cells by forced Math1 expression may be a cure for hearing loss. However, satisfying gene delivering vectors in gene therapy are not available. We combined quaternized chitosan (QCS) with Na-carboxymethyl-beta-cyclodextrin (CM-beta-CD) as novel non-viral vector, which adsorbs pRK5-Math1-EGFP perfectly at the mass ratio of 4:1. In vitro cell transfection can reach a 40% transfect efficiency and relatively low cytotoxity than liposomes. These results suggest that QCS/CM-beta-CD nanoparticle complexes could be a novel non-viral gene carrier in further clinical application.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Quitosana/química , Técnicas de Transferência de Genes , Nanopartículas , Amido/análogos & derivados , beta-Ciclodextrinas/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Amido/química
7.
Chin Med J (Engl) ; 123(13): 1633-8, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20819620

RESUMO

BACKGROUND: Our previous studies have shown that both apoptosis and necrosis are involved in hair cell (HC) pathogenesis in aging cochleae. To better understand the biological mechanisms responsible for the regulation of HC death, we examined the activity of succinate dehydrogenase (SDH), a mitochondrial bioenergetic enzyme, in the HCs of aging cochleae. METHODS: The auditory brainstem response thresholds elicited by tone bursts at 4, 10 and 20 kHz were measured in both young (2-3 months) and aging (22-23 months) Wistar rats. SDH activity was evaluated with a colorimetric assay using nitroblue tetrazolium monosodium salt. The SDH-labeled organs of Corti were double stained with propidium iodide, a DNA intercalating fluorescent probe for illustration of HC nuclei. All the specimens were examined with fluorescence microscopy and confocal microscopy. RESULTS: Aging rats exhibited a significant elevation of ABR thresholds with threshold shifts being 34 dB at 20 kHz, 28 dB at 10 kHz, and 25 dB at 4 kHz. Consistent with the reduction in the cochlear function, aging cochleae exhibited the reduction of SDH staining intensity in the apical and the basal ends of the cochleae, where a large number of apoptotic, necrotic, and missing HCs were evident. The reduction in SDH staining appeared in a cell-death-mode dependent fashion. Specifically, SDH labeling remained in apoptotic HCs. In contrast, SDH staining was markedly reduced or absent in necrotic HCs. CONCLUSIONS: In the aging cochlea, SDH activity is preserved in HCs undergoing apoptosis, but is substantially reduced in necrosis. These results suggest that mitochondrial energetic function is involved in the regulation of cell death pathways in the pathogenesis of aging cochleae.


Assuntos
Cóclea/citologia , Cóclea/enzimologia , Células Ciliadas Auditivas/enzimologia , Succinato Desidrogenase/metabolismo , Envelhecimento/metabolismo , Animais , Apoptose/fisiologia , Feminino , Masculino , Necrose/fisiopatologia , Ratos , Ratos Wistar , Succinato Desidrogenase/genética
8.
Dev Neurobiol ; 69(2-3): 153-61, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19067324

RESUMO

The Smads are a group of related intracellular proteins critical for transmitting the signals to the nucleus from the transforming growth factor-beta superfamily at the cell surface. Knockout of the Smad5 is embryonic lethal. However, the Smad5 knockout of single allele (+/-) could survive. We used Smad5 heterozygous knockout (+/-) to determine the role of Smad5 in the development of inner ear morphology and function. In situ hybridization showed that Smad5 was expressed predominantly in hair cells, spiral ganglion, and supporting cells. Measurements of hearing thresholds using auditory brainstem response showed that Smad5 defect resulted in progressive hearing loss between 4 and 24 weeks after birth. Morphological examination revealed apoptosis in the inner ear, with significant loss of outer hair cells in adult Smad5 mutant mice. Our results indicated that deficiency in the Smad5-mediated signaling resulted in apoptosis of hair cells, suggesting Smad5 is a gene that may be related with presbycusis.


Assuntos
Apoptose/genética , Cóclea/patologia , Células Ciliadas Auditivas/patologia , Perda Auditiva/genética , Proteína Smad5/deficiência , Proteína Smad5/metabolismo , Estimulação Acústica/métodos , Fatores Etários , Animais , Animais Recém-Nascidos , Limiar Auditivo/fisiologia , Cóclea/crescimento & desenvolvimento , Cóclea/ultraestrutura , Células Ciliadas Auditivas/ultraestrutura , Marcação In Situ das Extremidades Cortadas/métodos , Camundongos , Camundongos Knockout , Microscopia Eletrônica/métodos
9.
Artigo em Chinês | MEDLINE | ID: mdl-18717312

RESUMO

OBJECTIVE: To investigate the effect of Hath1 (human atonal homolog 1) overexpression on greater epithelial ridge (GER) cells from postnatal rat cochlea in vitro. METHODS: GER cells were isolated by using a combinatorial approach of enzymatic digestion and mechanical separation from P1 rat cochlear. The GER cell cultures were infected by adenovirus containing Hath1 and enhanced green fluorescent protein (ad-Hath1-EGFP), while transfecting EGFP(ad-EGFP) was as controls. Immunostaining were performed at different time points after adenovirus infection. RESULTS: Some of the infected GER cells became myosin VIIa-positive following ad-Hath1-EGFP infection. The earliest time point to see induction of hair cell differentiation (hair cell marker expression) by ad-Hathl was 5 days post-infection. In contrast, infection of the GER sheet cultures with ad-EGFP control virus did not show any myosin VIIa-positive cells at 3-12 days post-infection in all cultures examined. CONCLUSIONS: GER cells may potentially serve as hair cell progenitors and they are capable of differentiating hair cell-like cells when forced to express Hath1.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Cóclea/citologia , Células Epiteliais/citologia , Transfecção , Animais , Animais Recém-Nascidos , Diferenciação Celular , Células Cultivadas , Células Ciliadas Auditivas/citologia , Ratos , Ratos Sprague-Dawley
10.
Artigo em Chinês | MEDLINE | ID: mdl-17886676

RESUMO

OBJECTIVE: To analyze the molecular pathogenesis of deaf couples by means of genetic testing. To provide accurate genetic counseling and instruction for deaf couples with different etiology based upon results of genetic testing. METHODS: Four deaf families from July 2005 to May 2006. Each subject was with moderate to profound hearing loss. Genomic and mitochondrial DNA (mtDNA) of each subject were extracted from whole blood. Genetic testing of GJB2, SLC26A4 (PDS) and mtDNA A1555G mutation were offered to each individuals. RESULTS: The husband from family 1 didn't carry GJB2, SLC26A4 and mtDNA A1555G mutation while his wife was confirmed to carry compound SLC26A4 mutations. The possibility of their offspring's to be SLC26A4 single mutation carrier was 100%. The couple from family 2 both didn't carry GJB2, SLC26A4 and mtDNA A1555G mutation. The possibility of their offspring's having hereditary deafness caused by GJB2, SLC26A4 and mtDNA A1555G mutation was excluded. The husband from family 3 was confirmed to carry homozygous GJB2 mutations and a single SLC26A4 mutation while his wife who was diagnosed with enlarged vestibular aqueduct syndrome (EVAS) by CT scan was proven to carry a single SLC26A4 mutation. The risk of their offspring's suffering EVAS was 50%. The husband from family 4 was mtDNA A1555G positive while his wife who was diagnosed with cochlear malformation by CT scan didn't carry GJB2, SLC26A4 and mtDNA A1555G mutation. The risk of their offspring's having hereditary deafness caused by GJB2, SLC26A4 and mtDNA A1555G mutation was excluded. CONCLUSIONS: Genetic testing could be applied to offer the more accurate genetic counseling and instruction to deaf couples.


Assuntos
Surdez/genética , Surdez/prevenção & controle , Aconselhamento Genético , Conexina 26 , Conexinas/genética , DNA Mitocondrial/análise , Surdez/diagnóstico , Feminino , Doenças Genéticas Inatas/diagnóstico , Doenças Genéticas Inatas/genética , Genótipo , Humanos , Masculino , Proteínas de Membrana Transportadoras/genética , Mutação , Transportadores de Sulfato
11.
Zhonghua Yi Xue Za Zhi ; 87(16): 1088-92, 2007 Apr 24.
Artigo em Chinês | MEDLINE | ID: mdl-17672986

RESUMO

OBJECTIVE: To analyze the molecular genetic mechanisms of pathogenesis of deafness in the families with deaf-mute patients and analyze the strategies of genetic counseling and intervention for these families. METHODS: Peripheral blood samples were collected from the probands with deaf-muteness and their parents of five families and genetic tests were conducted to analyze the GJB2, SLC26A4 (PDS), and mitochondrial DNA (mtDNA) A 1555G genes for the existence of mutation. Families 1-3 had one child with hearing loss each while the parents had normal hearing and the mothers had been pregnant for 6-18 weeks. Both parents of family 4 were deaf-mute, and the wife of family 5 was deaf-mute while her husband had normal hearing. RESULTS: The proband from family 1 was proven to carry compound GJB2 mutations while his parents carried a single GJB2 mutation; prenatal testing showed that the fetus only carried the paternal mutation. The proband from family 2 was proven to carry compound SLC26A4 (PDS) mutations while his parents carried a single SLC26A4 (PDS) mutation; prenatal testing showed that the fetus only carried the paternal mutation. The proband from family 3 and his parents didn't carry any GJB2, SLC26A4 and mtDNA A1555G mutation. Observation showed that the new born babies of these three families all had normal hearing revealed by new born hearing screening and ABR test. The husband from family 4 was homozygous GJB2 235delC while his wife was mtDNA A1555G positive. This couple was advised to strictly avoid the administration of aminoglycoside antibiotics to their future offspring. In family 5, the wife carried compound SLC26A4 (PDS) mutations while her husband carried a single SLC26A4 (PDS) mutation; and they were told about the 50% risk of their offspring's suffering from enlarged vestibular aqueduct syndrome. CONCLUSION: Genetic testing with prenatal testing and relevant intervention for the families with deaf-mute patients can be applied to prevent another deaf-mute member from being born.


Assuntos
Conexinas/genética , Aconselhamento Genético , Perda Auditiva/genética , Mutação , Adulto , Sequência de Bases , Criança , Conexina 26 , Análise Mutacional de DNA , Saúde da Família , Feminino , Testes Genéticos , Perda Auditiva/diagnóstico , Perda Auditiva/prevenção & controle , Humanos , Masculino , Linhagem , Gravidez , Primeiro Trimestre da Gravidez , Segundo Trimestre da Gravidez , Diagnóstico Pré-Natal/métodos
12.
J Neurosci Methods ; 164(2): 271-9, 2007 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-17583357

RESUMO

Mammalian cochlear hair cell loss is irreversible and leads to permanent hearing loss. To restore hearing physiologically, it is necessary to generate new functional hair cells either from endogenous cells or from exogenously transplanted hair cells/progenitors. Previous studies suggest that cochlear greater epithelial ridge (GER) and lesser epithelial ridge (LER) cells are capable of differentiating into hair cells. While it was recently possible to obtain and culture pure LER progenitors, isolation of pure GER progenitors has not been reported. Here we describe a method that allows isolation of pure GER cells from neonatal rat cochleae. The cochlear epithelial sheet (CES) containing GER progenitor cells was mechanically separated from the underlying mesenchymal tissue after digestion with thermolysin. The GER area could then be dissected following mechanical removal of organ of Corti as well as all the lateral area. The isolated GER cells showed significant proliferation and expressed markers for GER cells but not markers for hair cells or LER. When the GER cells were cultured in serum-free medium containing epidermal growth factor, spheres were formed where they continued to proliferate. Furthermore, when GER cells were induced to express Hath1 or co-cultured with mesenchymal cells prepared from neonate rat cochleae, they showed the potential to differentiate into hair cell-like cells. Successful isolation, culture and differentiation of GER hair cell progenitors will shed additional light on the mechanism of hair cell differentiation and potential hair cell replacement.


Assuntos
Diferenciação Celular/fisiologia , Proliferação de Células , Separação Celular/métodos , Cóclea/citologia , Células Ciliadas Auditivas/fisiologia , Células-Tronco/fisiologia , Animais , Animais Recém-Nascidos , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Células Cultivadas , Dineínas/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Células Ciliadas Auditivas/ultraestrutura , Microscopia Eletrônica de Varredura , Miosina VIIa , Miosinas/metabolismo , RNA Mensageiro/metabolismo , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Células-Tronco/ultraestrutura , Transfecção/métodos
13.
Zhonghua Yan Ke Za Zhi ; 43(1): 55-8, 2007 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-17442165

RESUMO

OBJECTIVE: Transducing PEDF-GFP plasmid to the retina of the BN rats with cationic liposome through different gene delivery route, then observe the expression and location of the PEDF-GFP. METHODS: PEDF-GFP plasmid with cationic liposome was delivered to the retina of the BN rat through subretinal injection and intravitreal injection. The expression of GFP was observed under fluorescence microscope, and the mRNA of PEDF gene was detected by RT-PCR. RESULTS: Green fluorescence was emitted from the total retina include RPE cell under fluorescent microscope after 24 h in two gene delivery route, The fluorescence intensity was stronger with time changing. Gradual fluorescence increase in the retina and RPE cells occurred and lasted 4 weeks. The expression of PEDF mRNA was also detected by RT-PCR after 24 h, and maintained stable 4 weeks after infection. CONCLUSIONS: Cationic liposome can mediate PEDF-GFP gene into the retina of the BN rat effectively; subretinal injection and intravitreal injection both are effective gene delivery route; and their stable expression can maintain 4 weeks after transfection.


Assuntos
Receptores de Neuropeptídeos/genética , Retina , Epitélio Pigmentado da Retina , Transfecção , Animais , Vetores Genéticos , Proteínas de Fluorescência Verde/genética , Lipossomos , Masculino , Plasmídeos , Ratos , Ratos Endogâmicos BN , Epitélio Pigmentado da Retina/citologia
14.
Artigo em Chinês | MEDLINE | ID: mdl-18229588

RESUMO

OBJECTIVE: To establish in vitro culture systems of greater epithelial ridge (GER) cells from rat cochlear and to investigate the characterization, growth pattern and ultrastructure of GER cells. METHODS: Using a combinatorial approach of enzymatic digestion and mechanical separation to allow isolation and culture of GER cells from P1 rat cochleae. The dissociated GER cells were cultured in serum-free or 10% fetal bovine serum DMEM respectively. BrdU, phalloidin, ZO1, calretinin and myosin VIIa immunostaining and scanning electron microscope observation were performed in GER cell cultures. RESULTS: The dissociated GER cell cultures showed positive to ZO1, phalloidin and BrdU staining, but negative to myosin VIIa and calretinin. They assumed a polygonal morphology which was similar to epithelial cells and grew in islands-like patches in medium containing 10% fetal bovine serum while forming spheres in serum-free medium. The GER cells presented significant ability to proliferate in both conditions. Scanning electron microscope showed that there was microvillus and centre bodies but not hair cell specific stereociliary bundles on the surface of GER cultures. CONCLUSIONS: The GER cell cultures showed significant ability to proliferate and grew in islands-like patches in medium containing 10% fetal bovine serum while forming spheres in serum-free medium. The dissociated GER cells expressed epithelial cell specific marker but not marker of hair cells.


Assuntos
Técnicas de Cultura de Células , Cóclea/citologia , Células Epiteliais/citologia , Animais , Animais Recém-Nascidos , Células Cultivadas , Células Ciliadas Auditivas/citologia , Ratos , Ratos Sprague-Dawley
15.
Artigo em Chinês | MEDLINE | ID: mdl-17007371

RESUMO

OBJECTIVE: To investigate the incidence of hot spot mutation of PDS gene by genetic screening testing method in Chifeng City, Inner Mongolia. The feasibility and effectiveness of genetic screening method in finding enlarged vestibular aqueduct syndrome were confirmed by temporal bone CT scan. METHODS: DNA were extracted from peripheral blood of 141 students of Chifeng Deaf and Dumb school. PDS IVS7-2 A-G mutation, the most common PDS mutation in Chinese population, was analyzed by direct sequencing for PDS exon 7, exon 8 with intron 7. The individuals found with homozygous or heterozygous PDS IVS7-2 A-G mutation were given further temporal CT scan, ultrasound scan of thyroid and thyroid hormone assays. The results of PDS genetic screening and temporal bone CT scan were compared with each other. RESULTS: The sequencing results revealed twenty cases carrying PDS IVS7-2 A-G mutation, of whom nine cases were homozygous mutation and eleven cases were heterozygous mutation. Eighteen cases underwent temporal bone CT scan except two cases that left the school due to other health problem. Sixteen cases were confirmed to be enlarged vestibular aqueduct syndrome (EVAS) by CT scan and the shape and function of thyroid were clinically normal by ultrasound scan of thyroid and thyroid hormone assays, respectively. CONCLUSIONS: The patients suffered from EVAS can be diagnosed by the screening for the PDS hot spot mutation which has unique advantage in epidemiologic study in large scale deaf population. The preliminary data of this study suggested relatively high incidence of EVAS in Chifeng area.


Assuntos
Perda Auditiva/genética , Proteínas de Membrana Transportadoras/genética , Mutação Puntual , Aqueduto Vestibular/patologia , Doenças Vestibulares/genética , Adolescente , Criança , Pré-Escolar , China , Feminino , Testes Genéticos , Humanos , Transportadores de Sulfato , Síndrome , Adulto Jovem
16.
Artigo em Chinês | MEDLINE | ID: mdl-16335396

RESUMO

OBJECTIVE: To detect the expression of Math1, Hes1 and Hes5 in greater epithelial ridge (GER) cells of rat cochlear and explore their influence on hair cell differentiation. METHODS: Postnatal day 0 (P0), day 1 (P1) , day 3 (P3) day 4 (P4) and day 5 (P5) rat cochlear were dissected respectively and then pure GER cells were separated by a combinatorial approach of attachment and mechanical separation. The total RNA of GER cells was extracted by Trizol one step method and the expression of Math1, Hes1 and Hes5 in GER cells was detected with reverse transcription polymerase chain reaction. RESULTS: Math1 was expressed in P0 - P5 rat GER cells and Hes1 was expressed only in PO - P3 rat GER cells, while there was no expression of Hes5 in P0 - P5 rat GER cells. CONCLUSIONS: Probably only when the expression of Math1 reaches a certain level can it induce GER cells to differentiate into hair cells. Meanwhile this process might controlled by Hes1 to some extent.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Cóclea/metabolismo , Células Epiteliais/metabolismo , Células Ciliadas Auditivas/metabolismo , Proteínas de Homeodomínio/metabolismo , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Diferenciação Celular , Cóclea/citologia , Regulação da Expressão Gênica no Desenvolvimento , Células Ciliadas Auditivas/citologia , Proteínas de Homeodomínio/genética , Ratos , Ratos Sprague-Dawley , Fatores de Transcrição HES-1
17.
Acta Otolaryngol ; 124(2): 124-9, 2004 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15072413

RESUMO

OBJECTIVE: To determine the protective effects of basic fibroblast growth factor (bFGF) on cochlear neurons and hair cells in vitro and in vivo. MATERIAL AND METHODS: In Experiment I, cultured spiral ganglion neurons (SGNs) prepared from postnatal Day 3 mice were exposed to 20 mM glutamate for 2 h before the culture medium was replaced with fresh medium containing 0, 25, 50 or 100 ng/ml bFGF. Fourteen days later, all cultures were fixed with 4% paraformaldehyde and stained with 1% toluidine blue. The number of surviving SGNs was counted and the length of the neurites of the SGNs was measured. In Experiment II, in vivo studies were carried out with guinea pigs in which bFGF or normal saline was injected intramuscularly to assess possible protective effects of bFGF on cochlear hair cells and to accelerate the recovery of the auditory brainstem response (ABR). The ABRs were measured before, immediately after and 2 and 4 weeks after exposure to noise. RESULTS: Exposure to 20 mM glutamate for 2 h resulted in an inhibition of neurite outgrowth of SGNs and an increase in cell death. Treatment of the cultures with bFGF led to promotion of neurite outgrowth and an increase in the number of surviving SGNs. In Experiment II, significant (p < 0.05) differences in ABR thresholds were observed between the groups injected with bFGF and saline (t = 2.689) at 4 weeks after noise exposure. Cochleae were removed and hair cell loss analyzed in surface preparations prepared from all experimental animals. Acoustic trauma caused loss of 240 and 2160 inner hair cells in the groups injected with bFGF and saline, respectively. Similarly, more outer hair cells were lost in the normal saline injection group (99,291) than in the group treated with bFGF (70,377). CONCLUSIONS: Our results demonstrate that bFGF protects SGNs against glutamate neurotoxicity in vitro. In addition, treatment with bFGF protects hair cells from acoustic trauma.


Assuntos
Nervo Coclear/efeitos dos fármacos , Fator 2 de Crescimento de Fibroblastos/metabolismo , Células Ciliadas Auditivas/efeitos dos fármacos , Perda Auditiva Provocada por Ruído/prevenção & controle , Neurônios/efeitos dos fármacos , Síndromes Neurotóxicas/prevenção & controle , Animais , Audiometria de Resposta Evocada/métodos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Nervo Coclear/citologia , Ácido Glutâmico/toxicidade , Cobaias , Camundongos , Ruído/efeitos adversos , Gânglio Espiral da Cóclea/citologia
18.
Acta Otolaryngol ; 123(4): 552-4, 2003 May.
Artigo em Inglês | MEDLINE | ID: mdl-12797593

RESUMO

OBJECTIVE: In order to assess the reconstructive properties of fascia lata, superficial fascia lata and bone morphogenetic protein (BMP) in skull base surgery, lateral skull base bone and dura mater defect models were established in dogs MATERIAL AND METHODS: As a repair material we selected fascia lata, either alone or in combination with BMP, for reconstructing large cranial defects in dogs. Twenty dogs undergoing a 3.0 x 4.0 cm2 full-thickness excision of the parietal bone were divided into four equal groups as follows: fascia lata reconstruction; fascia lata reconstruction plus BMP; controls; and fascia lata reconstruction plus BMP with direct exposure of fascia lata. The implants were harvested at 2-15 weeks and examined histologically. Results-Treated and untreated implants were quite different: formation of new bone occurred in the dogs treated with BMP whereas the unreconstructed controls demonstrated only a bridge of fibrovascular connective tissue. CONCLUSION: The results of this study suggest that it is better to combine BMP and reconstructive material for the treatment of bone defects.


Assuntos
Proteínas Morfogenéticas Ósseas , Dura-Máter/cirurgia , Fascia Lata/cirurgia , Crânio/cirurgia , Animais , Bovinos , Cães , Feminino , Masculino , Procedimentos de Cirurgia Plástica , Soroalbumina Bovina
19.
Zhonghua Er Bi Yan Hou Ke Za Zhi ; 38(1): 21-3, 2003 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-12778761

RESUMO

OBJECTIVE: To construct basic fibroblast growth factor(bFGR) and enhance green fluorescence protein(EGFP) fusion gene eukaryotic expression vector internal ribosome entry site (pIRES)-bFGF-GFP and to evaluate the effect of transduction bFGF gene on noise induced hearing-loss in inner ear hair cells of guinea pigs. METHODS: Human bFGF cDNA was inserted into mammalian expressed plasmid pIRES-EGFP. The recombinant expression plasmid pIRES-bFGF-EGFP was transfected into inner ear of guinea pigs, using lipofectin method. The transduced bFGF gene was mediated by SA lipidsome. IRES- bFGF-GFP was administered into the round window as rescue agent at the same time of noise exposure or as a protective agent 7 days before. RESULTS: SA liposome-mediated bFGF expressed at a high level in the cochlea of guinea pigs, and in the rescue group, a significant lower hearing thresholds was displayed. pIRES- bFGF-EGFP could protect hair cells. It demonstrated that pIRES- bFGF-GFP could protect the inner ear both structurally and functionally. bFGF/EGFP gene could be transcripted and translated into inner ear hair cells of guinea pig. bFGF/EGFP gene could express a specific protein. The recombinant bFGF/EGFP had significant protective effect as well as manifestation of autonomous fluorescence. CONCLUSION: bFGF/EGFP fusion protein not only expressed in hair cells of guinea pig but also showed significant bFGF activity and autonomous fluorescence. IRES induced exogenous gene could enter the hair cells.


Assuntos
Fator 2 de Crescimento de Fibroblastos/genética , Vetores Genéticos , Perda Auditiva Provocada por Ruído/terapia , Animais , Feminino , Fator 2 de Crescimento de Fibroblastos/fisiologia , Expressão Gênica , Terapia Genética , Cobaias , Células Ciliadas Auditivas , Humanos , Masculino
20.
Acta Otolaryngol ; 122(4): 370-3, 2002 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12125991

RESUMO

The purpose of this study was to observe the protective effects of basic fibroblast growth factor (bFGF) on the cells of the inner ear using in vivo experiments. The studies were carried out using guinea pigs in which bFGF or artificial perilymph was perfused into the cochlea. The compound action potential (CAP) was measured before and after exposure to a sound simulating an explosion. The difference in CAP was significant between the bFGF-perfused group and the control group (p < 0.01, t = 3.896) and between the bFGF-perfused group and the artificial perilymph-perfused group (p < 0.05, t = 2.520). The cochleae were removed and hair cell loss estimated from surface preparations. Acoustic trauma caused loss of outer hair cells in the first and second turns of the cochlea in the bFGF-perfused group and the artificial perilymph-perfused group and partial loss of inner hair cells in the control group. Treatment with bFGF reduced the loss of inner hair cells compared to that of control animals. Our results demonstrate that treatment with bFGF protects the hair cells from acoustic trauma and may facilitate the recovery of hearing.


Assuntos
Fator 2 de Crescimento de Fibroblastos/farmacologia , Fator 2 de Crescimento de Fibroblastos/uso terapêutico , Células Ciliadas Auditivas/efeitos dos fármacos , Perda Auditiva Provocada por Ruído/tratamento farmacológico , Animais , Cobaias
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