Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 17 de 17
Filtrar
Mais filtros

Base de dados
País/Região como assunto
Tipo de documento
País de afiliação
Intervalo de ano de publicação
1.
Proc Natl Acad Sci U S A ; 118(22)2021 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-34050020

RESUMO

Genes that are primarily expressed in cochlear glia-like supporting cells (GLSs) have not been clearly associated with progressive deafness. Herein, we present a deafness locus mapped to chromosome 3p25.1 and an auditory neuropathy spectrum disorder (ANSD) gene, TMEM43, mainly expressed in GLSs. We identify p.(Arg372Ter) of TMEM43 by linkage analysis and exome sequencing in two large Asian families segregating ANSD, which is characterized by inability to discriminate speech despite preserved sensitivity to sound. The knock-in mouse with the p.(Arg372Ter) variant recapitulates a progressive hearing loss with histological abnormalities in GLSs. Mechanistically, TMEM43 interacts with the Connexin26 and Connexin30 gap junction channels, disrupting the passive conductance current in GLSs in a dominant-negative fashion when the p.(Arg372Ter) variant is introduced. Based on these mechanistic insights, cochlear implant was performed on three subjects, and speech discrimination was successfully restored. Our study highlights a pathological role of cochlear GLSs by identifying a deafness gene and its causal relationship with ANSD.


Assuntos
Códon sem Sentido , Conexinas/metabolismo , Genes Dominantes , Perda Auditiva Central/genética , Proteínas de Membrana/genética , Animais , Implante Coclear , Feminino , Perda Auditiva Central/metabolismo , Perda Auditiva Central/fisiopatologia , Perda Auditiva Central/cirurgia , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Linhagem , Percepção da Fala
2.
Eur Arch Otorhinolaryngol ; 277(12): 3331-3339, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32447495

RESUMO

OBJECTIVES: The identification of gene mutations enables more appropriate genetic counseling and proper medical management for EVA patients. The purpose of this study was to validate the accuracy and sensitivity of our method for comprehensive mutation detection in EVA, and summarize these data to explore a more accurate and convenient genetic diagnosis method. METHODS: A multiplex PCR sequencing panel was designed to capture the exons of three known EVA-associated genes (SLC26A4, KCNJ10, and FOXI1), and NGS was conducted in 17 Chinese families with EVA. RESULTS: A total of 16 SLC26A4 variants were found in 21 probands with bilateral EVA, including three novel variants (c.416G>A, c.823G>A and c.1027G>C), which were not reported in the dbSNP, gnomAD database, and ClinVar databases. One patient carried a FOXI1 variant (heterozygous, c.214C>A) and one patient carried a KCNJ10 variant (heterozygous, c.1054C>A), both of which were novel variants. Biallelic potential pathogenic variants were detected in 21/21patient samples, leading to a purported diagnostic rate of 100%. All results were verified by Sanger sequencing. CONCLUSION: This result supplemented the mutation spectrum of EVA, and supports that combined multiple PCR-targeted enrichment, and NGS is a valuable molecular diagnostic tool for EVA, and is suitable for clinical application.


Assuntos
Fatores de Transcrição Forkhead/genética , Perda Auditiva Neurossensorial/genética , Perda Auditiva/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Mutação/genética , Canais de Potássio Corretores do Fluxo de Internalização/genética , Transportadores de Sulfato/genética , Aqueduto Vestibular/anormalidades , Adolescente , Povo Asiático/genética , Criança , Pré-Escolar , Análise Mutacional de DNA , Éxons , Feminino , Perda Auditiva Neurossensorial/etnologia , Heterozigoto , Humanos , Masculino , Proteínas de Membrana Transportadoras/genética , Reação em Cadeia da Polimerase Multiplex , Adulto Jovem
3.
Hum Genet ; 137(4): 329-342, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29713870

RESUMO

Autosomal dominant nonsyndromic hearing loss (ADNSHL) is a highly genetically heterogeneous disorder. Up to date only approximately 37 ADNSHL-causing genes have been identified. The goal of this study was to determine the causative gene in a five-generation Chinese family with ADNSHL. A Chinese family was ascertained. Simultaneously, two affected individuals and one normal hearing control from the family were analyzed by whole exome capture sequencing. To assess the functional effect of the identified variant, in-vitro studies were performed. novel missense variant, c.512A>G (p.His171Arg) in exon 8 of the ELMO domain-containing 3 (ELMOD3) gene, was identified as a causative variant in this family affected by late-onset and progressive ADNSHL. The variant was validated by Sanger sequencing and found to co-segregate with the phenotype within the pedigree and was absent in 500 ethnically matched unrelated normal hearing control subjects. To our knowledge, this is the first report of a family with ADNSHL caused by ELMOD3 mutation. Western blots and immunofluorescence staining demonstrated that p.His171Arg resulted in abnormal expression levels of ELMOD3 and abnormal subcellular localization. Furthermore, the analysis of the stability of the wild-type (WT) and mutant ELMOD3 protein shows that the decay of p.His171Arg is faster than that of the WT, suggesting a shorter halflife of the c.512A > G variant. A novel variant in the ELMOD3 gene, encoding a member of the engulfment and cell motility (ELMO) family of GTPase-activating proteins, was identified for the first time as responsible for ADNSHL.


Assuntos
Proteínas Ativadoras de GTPase/genética , Perda Auditiva Neurossensorial/genética , Adulto , Sequência de Aminoácidos/genética , Movimento Celular/genética , China/epidemiologia , Exoma/genética , Feminino , Perda Auditiva Neurossensorial/fisiopatologia , Humanos , Masculino , Mutação , Linhagem , Fenótipo
4.
J Hum Genet ; 63(6): 723-730, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29559740

RESUMO

X-linked inheritance is very rare and is estimated to account for only 1-5% of all nonsyndromic hearing loss cases. We found a multiplex family from China segregating with X-linked nonsyndromic hearing loss. After exclusive analysis of 10 common variations of three hearing loss-related genes, GJB2, mtDNA12srRNA and SLC26A4, a novel truncated variant of SMPX, c.87dupA (p.Gly30Argfs*12) (NCBI ClinVar Submission ID: SUB3136126), was identified by whole-exome sequencing. This variant was co-segregated with hearing loss in the entire family and was absent in 576 unrelated ethnically and geographically matched controls. We also detected a single nucleotide variation in two male controls with normal hearing, SMPX c.55A>G (p.Asn19Asp), which has been annotated as a rare variant in the Single Nucleotide Polymorphism (dbSNP) (rs759552778) and Exome Aggregation Consortium (ExAC) databases. This study has enriched the mutation spectrum of the SMPX gene.


Assuntos
Doenças Genéticas Ligadas ao Cromossomo X/genética , Perda Auditiva Neurossensorial/genética , Proteínas Musculares/genética , Mutação , Adulto , Povo Asiático/genética , Estudos de Casos e Controles , China , Bases de Dados Genéticas , Feminino , Perda Auditiva Neurossensorial/etnologia , Humanos , Masculino , Linhagem , Polimorfismo de Nucleotídeo Único , Sequenciamento do Exoma , Adulto Jovem
5.
Cell Physiol Biochem ; 35(2): 419-32, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25613036

RESUMO

BACKGROUND: Metabotropic glutamate receptors (mGluRs) are G-protein-coupled receptors that mediate neuronal excitability and synaptic plasticity in the central nervous system, and emerging evidence suggests a role of mGluRs in the biology of cancer. Previous studies showed that mGluR1 was a potential therapeutic target for the treatment of breast cancer and melanoma, but its role in human glioma has not been determined. METHODS: In the present study, we investigated the effects of mGluR1 inhibition in human glioma U87 cells using specific targeted small interfering RNA (siRNA) or selective antagonists Riluzole and BAY36-7620. The anti-cancer effects of mGluR1 inhibition were measured by cell viability, lactate dehydrogenase (LDH) release, TUNEL staining, cell cycle assay, cell invasion and migration assays in vitro, and also examined in a U87 xenograft model in vivo. RESULTS: Inhibition of mGluR1 significantly decreased the cell viability but increased the LDH release in a dose-dependent fashion in U87 cells. These effects were accompanied with the induction of caspase-dependent apoptosis and G0/G1 cell cycle arrest. In addition, the results of Matrigel invasion and cell tracking assays showed that inhibition of mGluR1 apparently attenuated cell invasion and migration in U87 cells. All these anti-cancer effects were ablated by the mGluR1 agonist L-quisqualic acid. The results of western blot analysis showed that mGluR1 inhibition overtly decreased the phosphorylation of PI3K, Akt, mTOR and P70S6K, indicating the mitigated activation of PI3K/Akt/mTOR pathway. Moreover, the anti-tumor activity of mGluR1 inhibition in vivo was also demonstrated in a U87 xenograft glioma model in athymic nude mice. CONCLUSION: The remarkable efficiency of mGluR1 inhibition to induce cell death in U87 cells may find therapeutic application for the treatment of glioma patients.


Assuntos
Antineoplásicos/administração & dosagem , Neoplasias Encefálicas/tratamento farmacológico , Glioma/tratamento farmacológico , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Receptores de Glutamato Metabotrópico/antagonistas & inibidores , Animais , Antineoplásicos/farmacologia , Neoplasias Encefálicas/metabolismo , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Glioma/metabolismo , Humanos , Camundongos , Camundongos Nus , Terapia de Alvo Molecular , Naftalenos/administração & dosagem , Naftalenos/farmacologia , Ácido Quisquálico/farmacologia , RNA Interferente Pequeno/administração & dosagem , RNA Interferente Pequeno/farmacologia , Receptores de Glutamato Metabotrópico/metabolismo , Riluzol/administração & dosagem , Riluzol/farmacologia , Ensaios Antitumorais Modelo de Xenoenxerto
6.
Biochem Biophys Res Commun ; 443(1): 138-43, 2014 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-24284040

RESUMO

Glutamate-mediated excitotoxicity is involved in many acute and chronic brain diseases. Dynamin related protein 1 (Drp-1), one of the GTPase family of proteins that regulate mitochondrial fission and fusion balance, is associated with apoptotic cell death in cancer and neurodegenerative diseases. Here we investigated the effect of downregulating Drp-1 on glutamate excitotoxicity-induced neuronal injury in HT22 cells. We found that downregulation of Drp-1 with specific small interfering RNA (siRNA) increased cell viability and inhibited lactate dehydrogenase (LDH) release after glutamate treatment. Downregulation of Drp-1 also inhibited an increase in the Bax/Bcl-2 ratio and cleavage of caspase-9 and caspase-3. Drp-1 siRNA transfection preserved the mitochondrial membrane potential (MMP), reduced cytochrome c release, enhanced ATP production, and partly prevented mitochondrial swelling. In addition, Drp-1 knockdown attenuated glutamate-induced increases of cytoplasmic and mitochondrial Ca(2+), and preserved the mitochondrial Ca(2+) buffering capacity after excitotoxicity. Taken together, these results suggest that downregulation of Drp-1 protects HT22 cells against glutamate-induced excitatory damage, and this neuroprotection may be dependent at least in part on the preservation of mitochondrial function through regulating intracellular calcium homeostasis.


Assuntos
Apoptose/fisiologia , Cálcio/metabolismo , Dinaminas/metabolismo , Ácido Glutâmico/metabolismo , Mitocôndrias/fisiologia , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular , Regulação para Baixo , Dinaminas/genética , Ácido Glutâmico/farmacologia , Camundongos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , RNA Interferente Pequeno/genética
7.
Front Neurosci ; 18: 1353759, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38327847

RESUMO

Objective: Anxiety symptoms are prevalent neuropsychiatric manifestations in Parkinson's disease (PD) and impact the development of motor complications. Our aim was to evaluate the association of GBA variants with the anxiety development in early PD cohort. Methods: This cohort study used data from the Parkinson Progression Marker Initiative. The primary outcome anxiety was assessed by State-Trait Anxiety Inventory (STAI). The association between GBA and longitudinal change in the STAI total score was examined using linear mixed-effects model, and the association between GBA and anxiety progression was examined using Cox survival analysis. Results: A total of 385 patients with PD were included in this study, 39 of them were GBA variant carriers and 346 were idiopathic PD without GBA variants. Patients with GBA variants had faster annual increase in anxiety score (ß = 0.44; 95% CI, 0.18 to 0.71; p < 0.001) and were at higher risk of anxiety progression (HR 1.87; 95% CI, 1.03 to 3.41; p = 0.03,). Higher baseline scores for Scales for Outcomes in Parkinson's Disease-Autonomic (SCOPA-AUT), which indicated the autonomic dysfunction, also independently predicted faster increase in anxiety score (ß = 0.48; 95%CI, 0.19 to 0.69; p < 0.001) and higher incidence of anxiety development (HR = 1.05; 95% CI, 1.01 to 1.08; p = 0.008). Interpretation: These findings suggest that longitudinal anxiety symptoms worsening was faster in PD patients who were GBA variant carriers and have dysautonomia, and this association was enhanced if they have both.

8.
J Neurol ; 270(5): 2756-2764, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36790548

RESUMO

BACKGROUND: Freezing of gait (FOG) is a common disabling gait disturbance among patients with Parkinson's disease (PD), but the influence of genetic variants on the incidence of FOG has been poorly studied to date. OBJECTIVES: We aimed to evaluate the association of GBA variants with the risk of FOG development in a large early PD cohort. METHODS: This study included 371 early PD patients from the Parkinson's Progression Markers Initiative (PPMI) who were divided into a GBA variant carrier group (GBA-PD group, n = 44) and an idiopathic PD group without GBA variants (iPD group, n = 327). They were followed up for up to 5 years to examine the progression of FOG. The cumulative incidence of FOG and risk factors for FOG were assessed using Kaplan‒Meier and Cox regression analyses. RESULTS: At baseline, the GBA-PD group had lower CSF ß-amyloid 1-42 (Aß42) levels and more severe motor and nonmotor symptoms than the iPD group. During the 5-year follow-up, the GBA-PD group had a higher incidence of FOG than the iPD group, and the FOG progression rate was related to GBA variant severity. In the multivariable Cox model without CSF Aß42, GBA variants were significant predictors of future FOG, and the association remained significant after adding CSF Aß42 to the model. In the subgroup analyses, the effect of GBA variants was not observed in the "low-level" group. However, in the "high-level" group, GBA variants independently increased the risk of FOG, and this association was stronger than the association with CSF Aß42. CONCLUSION: GBA variants are novel genetic risk factors for future FOG development in early PD patients. This association seemed to be mediated by both Aß-dependent pathways and Aß-independent pathways.


Assuntos
Transtornos Neurológicos da Marcha , Doença de Parkinson , Humanos , Marcha , Transtornos Neurológicos da Marcha/genética , Transtornos Neurológicos da Marcha/complicações , Incidência , Doença de Parkinson/complicações , Doença de Parkinson/genética , Doença de Parkinson/metabolismo , Fatores de Risco
9.
Front Neurosci ; 16: 832550, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35310107

RESUMO

Background and Purpose: Rapid eye movement (REM) Rapid eye movement sleep behavior disorder (RBD) is a common non-motor symptom of PD. However, the association between the SNCA rs3910105 genotype and RBD in Parkinson's disease (PD) remains unclear. Methods: This study used Parkinson's Progression Markers Initiative (PPMI) data and included 270 patients with newly diagnosed PD without RBD who were divided into SNCA rs3910105 C carriers (CC+CT; n = 187) and TT carriers (n = 83). They were followed up for 5 years to identify the development of RBD. To investigate the influence of cerebrospinal fluid (CSF) alpha-synuclein (α-syn) and ß-amyloid 1-42 (Aß42) in the association between rs3910105 and RBD, the patients were additionally classified into "high-level" and "low-level" groups using cutoff values for CSF α-syn and Aß42 levels. Results: At baseline, the rs3910105 C allele group had lower CSF α-syn and Aß42 levels than the TT group. During the 5.0-year follow-up, the rs3910105 C allele group had a higher incidence of RBD than the TT group. In the subgroup analyses, the effect of the rs3910105 C allele was not found in the "low-level" group. However, in the "high-level" group, the rs3910105 C allele independently increased the risk of RBD. Conclusion: The SNCA rs3910105 C allele might be a novel genetic risk factor for RBD development in PD, α-syn pathways might have a role in this association and more basic research would be needed to elucidate the mechanism in the future.

10.
Artigo em Zh | MEDLINE | ID: mdl-32086922

RESUMO

Objective:To detect 20 common deafness gene mutations in non- syndromic deafness patients in China using PCR- RDB, and analyze and summarize the mutation data to explore the clinical value of this method. Method:The PCR- RDB and Sanger sequencing were used to detect 20 common mutations of four deafness genes(GJB2, GJB3, SLC26A4 and mtDNA) in 500 patients with non- syndromic hearing loss . The Sanger sequencing was used to compare the sensitivity, specificity, positive predictive value, negative predictive value, and total coincidence rate of the deafness mutation detected by PCR- RDB. Result:A total of 500 samples were detected. 147 wild- type samples, 81 homozygous mutant samples, 240 heterozygous mutant samples, 32 composite heterozygous mutant samples were detected using the PCR- RDB within the range of 20 gene mutations, which were identical to the Sanger sequencing results. GJB2 c.235delC and SLC26A4 c.919- 2 A>G are the most common hotspot mutations in this study, followed by mtDNA m. 1555 A>G. Compared with the Sanger sequencing method, the sensitivity, specificity, positive predictive value, negative predictive value, and total coincidence rate of the real- time fluorescence PCR melting curve method were 100%, and the Kappa value was one. Conclusion:PCR reverse dot-blot hybridization is a simple, rapid, sensitive and specific method for detecting 20 mutations of 4 common deafness genes in Chinese population, it is expected to be used in clinical detection of deafness genes in the future.


Assuntos
Análise Mutacional de DNA , Surdez/genética , China , Conexina 26 , Conexinas/genética , DNA Mitocondrial/genética , Humanos , Mutação , Reação em Cadeia da Polimerase , Transportadores de Sulfato/genética
11.
Front Genet ; 10: 639, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31379920

RESUMO

Autosomal recessive non-syndromic hearing loss (ARNSHL) is a highly heterogeneous disease involving more than 70 pathogenic genes. However, most ARNSHL families have small-sized pedigrees with limited genetic information, rendering challenges for the molecular diagnosis of these patients. Therefore, we attempted to establish a strategy for identifying deleterious variants associated with ARNSHL by applying proband whole-exome sequencing (proband-WES). Aside from desiring to improve molecular diagnostic rates, we also aimed to search for novel deafness genes shared by patients with similar phenotype, making up for the deficiency of small ARNSHL families. In this study, 48.5% (16/33) families were detected the pathogenic variants in eight known deafness genes, including 10 novel variants identified in TMPRSS3 (MIM 605551), MYO15A (MIM 602666), TMC1 (MIM 606706), ADGRV1 (MIM 602851), and PTPRQ (MIM 603317). Apart from six novel variants with a truncating effect (nonsense, deletion, insertion, and splice-site), four novel missense variants were not found in 200 unrelated control population by using Sanger sequencing. It is important to note that none of novel genes were shared across different pedigrees, indicating that a larger sample size might be needed. Proband-WES is a cost-effective and precise way of identifying causative variants in nuclear families with ARNSHL. This economical strategy may be appropriated as a clinical application to provide molecular diagnostics, genetic counseling, and individualized health maintenance measures for patients with ARNSHL at hearing clinics.

12.
Gene ; 704: 113-120, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-30974196

RESUMO

Usher syndrome (USH) is a clinically common autosomal recessive disorder characterized by retinitis pigmentosa (RP) and sensorineural hearing loss with or without vestibular dysfunction. In this study, we identified a Hunan family of Chinese descent with two affected members clinically diagnosed with Usher syndrome type 3 (USH3) displaying hearing, visual acuity, and olfactory decline. Whole-exome sequencing (WES) identified a nonsense variant in ABHD12 gene that was confirmed to be segregated in this family by Sanger sequencing and exhibited a recessive inheritance pattern. In this family, two patients carried homozygous variant in the ABHD12 (NM_015600: c.249C>G). Mutation of ABHD12, an enzyme that hydrolyzes an endocannabinoid lipid transmitter, caused incomplete PHARC syndrome, as demonstrated in previous reports. Therefore, we also conducted a summary based on variants in ABHD12 in PHARC patients, and in PHARC patients showing that there was no obvious correlation between the genotype and phenotype. We believe that this should be considered during the differential diagnosis of USH. Our findings predicted the potential function of this gene in the development of hearing and vision loss, particularly with regard to impaired signal transmission, and identified a novel nonsense variant to expand the variant spectrum in ABHD12.


Assuntos
Códon sem Sentido , Monoacilglicerol Lipases/genética , Síndromes de Usher/genética , Adulto , Idoso , Ataxia/genética , Ataxia/patologia , Catarata/genética , Catarata/patologia , Criança , Família , Feminino , Estudos de Associação Genética , Predisposição Genética para Doença , Genótipo , Homozigoto , Humanos , Masculino , Linhagem , Fenótipo , Polineuropatias/genética , Polineuropatias/patologia , Retinose Pigmentar/genética , Retinose Pigmentar/patologia , Síndromes de Usher/patologia , Sequenciamento do Exoma
13.
World Neurosurg ; 119: e262-e271, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30053568

RESUMO

OBJECTIVE: To assess the application of functional neuronavigation in surgeries of adult cerebral gliomas. METHODS: We performed a retrospective analysis of 375 cases of adult cerebral glioma patients who underwent microsurgical treatment between 2011 and 2017 in our department. Among them, 142 patients underwent surgery using functional neuronavigation (group A), and the other 233 patients were operated on without the help of functional neuronavigation (group B). For both groups, we categorized them into several subgroups according to the lesion locations. RESULTS: A significant difference in the gross total resection rate was observed between group A and group B (P = 0.001 for overall; P = 0.036 for EO area; and P = 0.004 for BBT area). The postoperative complication rate of group A was much lower than that of group B (P = 0.003 for overall; and P = 0.016 for BBT area). The postoperative 6-month Karnofsky Performance Scale score of all patients in group A was significantly higher than that of group B. Kaplan-Meier survival analyses showed significant increases in the median survival time of group A compared with that of group B (P < 0.001 for overall; P = 0.012 for EO area; P = 0.006 for BBT area), and the Cox proportional regression analysis estimated the hazard ratio of the functional neuronavigation to be 0.533, helping reduce the risk of death by 46.7%. CONCLUSIONS: This study confirmed that the application of neuronavigation in adult glioma surgery can improve postoperative quality of life and lengthen the survival time of patients, especially in cases involving the brainstem and the eloquent area.


Assuntos
Neoplasias Encefálicas/cirurgia , Glioma/cirurgia , Microcirurgia/métodos , Neuronavegação/métodos , Adolescente , Adulto , Neoplasias Encefálicas/diagnóstico por imagem , Feminino , Glioma/diagnóstico por imagem , Humanos , Processamento de Imagem Assistida por Computador , Estudos Longitudinais , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Análise de Sobrevida , Resultado do Tratamento , Adulto Jovem
14.
Int J Pediatr Otorhinolaryngol ; 115: 114-119, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30368370

RESUMO

OBJECTIVE: To evaluate the accuracy and validity of our protocol for prenatal diagnosis and genetic counseling in high-risk families at a clinic. METHODS: Fifteen unrelated families with recessive nonsyndromic hearing loss (NSHL) in their family history and a positive attitude towards prenatal diagnosis were recruited in the present study. According to genetic information for each family, Sanger sequencing, fluorescence polymerase chain reaction (PCR)-based congenital deafness gene detection kit and multiple PCR-based target gene capture and high-throughput sequencing were used. Genetic counseling was offered to all participating families by genetic counselors and otologists. Prenatal diagnosis was provided to families with detected pathogenic mutations and who were expected to participate in subsequent prenatal diagnosis. RESULTS: In this study, confirmed pathogenic mutations were detected in eight families, who were defined as high-risk families. These families all participated in prenatal diagnosis with positive attitudes. One novel variant (c.1687dupA) in the SLC264 gene was detected in a family. Through genetic counseling, the recurrence probability of NSHL in fetuses was 25% in six families, 0% in one family, and 50% in one family. The results of fetal DNA detection showed that one fetal variant was wild type, three were heterozygous mutations in SLC26A4, and one was a compound heterozygous mutation in SLC26A4. Two variants were heterozygous mutations in GJB2, and one was a homozygous mutation in GJB2. According to the test results for fetal DNA, prenatal diagnosis found that six fetuses had normal hearing, whereas two fetuses suffered from NSHL. After birth, six infants predicted to have normal hearing passed a newborn hearing screening test and two infants predicted to have NSHL were diagnosed with NSHL and received cochlear implants. CONCLUSION: Our protocol for prenatal diagnosis and genetic counseling provides detailed information that can assist couples in high-risk families in preparing for infant arrival and future family planning. For the affected neonates, prenatal diagnosis and genetic counseling achieve an "early screening, early diagnosis, early intervention" strategy.


Assuntos
Surdez/diagnóstico , Aconselhamento Genético/métodos , Diagnóstico Pré-Natal/métodos , Conexinas/genética , Análise Mutacional de DNA , Surdez/genética , Feminino , Audição , Testes Auditivos , Heterozigoto , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Homozigoto , Humanos , Lactente , Recém-Nascido , Masculino , Mutação , Linhagem , Reação em Cadeia da Polimerase/métodos , Gravidez
15.
J Am Soc Mass Spectrom ; 26(12): 2062-76, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26450359

RESUMO

Protein tyrosine nitration is involved in the pathogenesis of highly fatal astrocytomas, a type of brain cancer. To understand the molecular mechanisms of astrocytomas and to discover new biomarkers/therapeutic targets, we sought to identify nitroproteins in human astrocytoma tissue. Anti-nitrotyrosine immunoreaction-positive proteins from a high-grade astrocytoma tissue were detected with two-dimensional gel electrophoresis (2DGE)-based nitrotyrosine immunoblots, and identified with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Fifty-seven nitrotyrosine immunopositive protein spots were detected. A total of 870 proteins (nitrated and non-nitrated) in nitrotyrosine-immunopositive 2D gel spots were identified, and 18 nitroproteins and their 20 nitrotyrosine sites were identified with MS/MS analysis. These nitroproteins participate in multiple processes, including drug-resistance, signal transduction, cytoskeleton, transcription and translation, cell proliferation and apoptosis, immune response, phenotypic dedifferentiation, cell migration, and metastasis. Among those nitroproteins that might play a role in astrocytomas was nitro-sorcin, which is involved in drug resistance and metastasis and might play a role in the spread and treatment of an astrocytoma. Semiquantitative immune-based measurements of different sorcin expressions were found among different grades of astrocytomas relative to controls, and a semiquantitative increased nitration level in high-grade astrocytoma relative to control. Nitro-ß-tubulin functions in cytoskeleton and cell migration. Semiquantitative immunoreactivity of ß-tubulin showed increased expression among different grades of astrocytomas relative to controls and semiquantitatively increased nitration level in high-grade astrocytoma relative to control. Each nitroprotein was rationalized and related to the corresponding functional system to provide new insights into tyrosine nitration and its potential role in the pathogenesis of astrocytoma formation. Graphical Abstract ᅟ.


Assuntos
Astrocitoma/química , Neoplasias Encefálicas/química , Encéfalo/patologia , Eletroforese em Gel Bidimensional/métodos , Proteínas/química , Espectrometria de Massas em Tandem/métodos , Tirosina/análogos & derivados , Adolescente , Adulto , Idoso , Sequência de Aminoácidos , Astrocitoma/patologia , Western Blotting , Química Encefálica , Neoplasias Encefálicas/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Tirosina/análise
16.
Brain Res ; 1555: 1-9, 2014 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-24508577

RESUMO

Mood stabilizer valproic acid (VPA), a widely used antiepileptic drug that has been demonstrated neuroprotective effect against various insults through multiple signaling pathways. The role of VPA in traumatic brain injury (TBI) remains unclear. In the present study, we investigated the neuroprotective potency of VPA for protection against TBI in adult rats, focusing on studying signaling mediators of two well characterized pro-survival molecules, extracellular signal-regulated protein kinase (ERK) and Akt. We found that treatment of VPA after TBI significantly attenuated brain edema, reduced contusion volume and the rate of neuronal apoptosis. The treatment also partly blocked an increase in capase-3 activity. VPA markedly up-regulated the activity of ERK and Akt expression. Moreover, treatment with either PD98059, an ERK inhibitor and/or LY294002, an Akt inhibitor, attenuated the neuroprotection of VPA against TBI to varying degrees. Taken together, these results demonstrated that treatment with VPA after TBI could be neuroprotective via activation of ERK and Akt signaling pathways.


Assuntos
Lesões Encefálicas/tratamento farmacológico , Córtex Cerebral/enzimologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Fármacos Neuroprotetores/uso terapêutico , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ácido Valproico/uso terapêutico , Animais , Apoptose/efeitos dos fármacos , Lesões Encefálicas/enzimologia , Lesões Encefálicas/patologia , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/patologia , Masculino , Ratos , Ratos Sprague-Dawley , Transdução de Sinais
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA