Objective To investigate the impact of the dominant deafness point mutation p.D179N on the oli-gomeric equilibrium state of Connexin 26(Cx26).Methods The wild-type Cx26 fusion protein(Cx26-WT-GFP)and mutant fusion proteins(Cx26-D179N-GFP,Cx26-D179C-GFP)were expressed in HEK293F cells.By using Fluorescence-detection size-exclusion chromatography(FSEC)and size-exclusion chromatography(SEC)to analysis the oligomeric state of the target protein based on malecular weight under the condition of solubilization and purifica-tion respectively.Cryo-electron microscopy(Cryo-EM)single particle analysis(SPA)was conducted to analysis the target protein's oligomeric states based on the 2D classification morphology of the protein particles.Results In vitro,the wild-type Cx26 protein(Cx26-WT)is almost exclusively dodecameric.The deafness mutation p.D179N protein(Cx26-D179N)exists as both dodecamers and hexamers,whereas the artificial mutation p.D179C protein(Cx26-D179C)does not form dodecamers.Conclusion The dominant deafness mutation GJB2 p.D179N could weaken the ability of docking between hexameric proteins,which could affect the balance between hexamers and do-decamers.

Objective To develop and translate the Chinese listening self-efficacy questionnaire(C-LSEQ)and to test its reliability and validity.Methods A total of 172 subjects aged≥60 years with age-related hearing loss completed the C-LSEQ questionnaire via direct interviews.Pure tone hearing threshold test,Mandarin hearing in noise test,and the hearing handicap inventory for elderly-screening(HHIE-S)were evaluated.Twenty subjects were randomly selected from the original group to complete the C-LSEQ questionnaire in 2 weeks after the initial evaluation.Results ① Reliability:The Cronbach's a coefficients of the three sub-dimensions and the overall ques-tionnaire were all＞0.8,and the test-retest reliability coefficients of the three sub-dimensions and the overall ques-tionnaire were all＞0.9(P＜0.001).② Validity:The experts collectively evaluated the representativeness of the statements with good content validity.The convergent validity test showed that the composite reliability(CR)of the 2 sub-dimensions and the overall questionnaire were all＞0.7,and the average variance extracted(AVE)were all＞0.5.The CR value of the complex listening dimension was 0.655,and the AVE value was 0.937.The criterion validity test showed that the overall questionnaire and the three sub-dimensions of the C-LSEQ were significantly correlated with the pure-tone average,speech recognition thresholds in noise,and HHIE-S(P＜0.001).Conclusion The C-LSEQ exhibits stable structure,with good reliability and validity.It can be used to assess listen-ing self-efficacy in adults with hearing loss,especially in the elderly with age-related hearing loss.

Objective To investigate the damage mechanism of extracellular vesicles(EVs)derived from ves-tibular schwannoma(VS)on HEI-OC1 cells and the protective effect of the ferroptosis inhibitor ferrostatin-1(Fer-1).Methods Tumor tissues and clinical data were collected from four patients with stage Ⅱ or Ⅲ VS,classified as grade D according to the AAO-HNS hearing classification.Primary VS cells were extracted,and their conditioned supernatant was collected.EVs were isolated using ultracentrifugation and identified.HEI-OC1 cells were cultured in vitro and divided into three groups:the control group(no treatment),the EVs group(treated with 3000 parti-cles/cell VS-EVs for 24 hours),and the EVs+Fer-1 group(pretreated with 20 μmol/L Fer-1 for 2 hours followed by co-culture with 3000 particles/cell VS-EVs for 24 hours).Cell viability was assessed using the CCK-8 assay,re-active oxygen species(ROS)levels were quantified using the DCFH-DA fluorescent probe,and lipid peroxidation was evaluated using the BODIPY 581/591 C11 probe.Results Compared with the control group,the EVs group showed significantly reduced cell viability(P＜0.001)and increased levels of ROS(P＜0.001)and lipid peroxides(P＜0.001).However,the EVs+Fer-1 group exhibited significantly restored cell viability(P＜0.001)and re-duced levels of ROS and lipid peroxidation(P＜0.001).Conclusion VS-derived EVs disrupts redox homeostasis,promotes intracellular accumulation of lipid peroxides and ROS,and induces ferroptosis in HEI-OC1 cells.Fer-1 significantly alleviates VS-EVs-induced ferroptosis,thereby protecting HEI-OC1 cells from damage.

Objectives:To investigate the protective effect of nicotinamide adenine dinucleotide (NAD?) against noise-induced cochlear damage and preliminarily explore its underlying transcriptional and metabolic regulatory mechanisms.Methods:During the study period (January 2023-February 2025), an oxidative stress model was established using House Ear Institute-organ of Corti 1 (HEI-OC1) cells, and cell viability was assessed using the Cell Counting Kit-8 (CCK8) assay. Flow cytometry was employed to analyze cell apoptosis. A mouse model of noise-induced hearing loss was developed, and the mice were divided into three groups: a noise-exposed saline group, a noise-exposed NAD? intervention group, and a noise-free control group. Hearing protection effects were evaluated by auditory brainstem response (ABR) and immunofluorescence. Metabolomics and transcriptomics were used to analyze the regulatory effects of NAD +on transcription and metabolism in mouse cochlea. Enzyme-linked immunosorbent assay, quantitative real-time PCR, and western blot were used to verify the differential transcription and metabolic molecules and their functions. Data were statistically analyzed with GraphPad Prism 9.3.0. Results:NAD +at concentrations ranging from 10-80 μM effectively restored cell viability and reduced apoptosis induced by H?O? in HEI-OC1 cells. NAD? intervention significantly improved 16-32 kHz ABR thresholds after noise exposure ( P<0.05), reduced outer hair cell loss rates ( P<0.05), and attenuated ribbon synapse damage ( P<0.000 1). Metabolomics analysis revealed a significant downregulation in the glycerophospholipid metabolism pathway, with decreased levels of lysophosphatidic acid (LPA) and its related metabolites. ELISA results showed that LPA levels in the NAD? intervention group were significantly lower ( P<0.05). LPA inhibitor (ATX inhibitor 1) exhibited a cell protective effect similar to that of NAD?. Transcriptomics analysis indicated a significant upregulation of key genes related to potassium ion channels, such as Kcnq4. qPCR and Western blot further confirmed the significant upregulation of Kcnq4 and its encoded protein in the NAD? intervention group ( P<0.05). In the presence of the KCNQ4 inhibitor (ML252), the protective effect of NAD? was inhibited. Conclusions:NAD? exerts effective protective effects against noise-induced cochlear injury. Its protective mechanism may be closely related to the inhibition of LPA metabolic pathway and the up-regulation of KCNQ4 channel function.

Meniere's disease represents an idiopathic inner ear disorder characterized by endolymphatic hy-drops.Currently,the research methods for identifying genes associated with this disease mainly involve first-genera-tion sequencing and second-generation sequencing.This article reviews research on the genetic study of Meniere's disease,mainly summarizing the candidate genes with repeated pedigree validation in familial Meniere's disease,as well as those frequently reported in sporadic Meniere's disease.

Objective To study the hearing intervention effects of the Shokz condyle-stimulated headband bone-conduction hearing aid on patients with conductive,mixed,and sensorineural hearing loss and to explore its clinical application prospects.Methods A total of 55 patients with hearing loss(age 18～82)participated in the study.Among them,9 had conductive hearing loss,15 had sensorineural hearing loss,and 31 had mixed hearing loss.Their bilat-eral bone conduction pure tone thresholds at 0.5,1,2,and 4 kHz were all ≤60 dB HL.The patients were fitted with the condyle-stimulated headband bone-conduction hearing aid.Hearing thresholds in sound field,single-syllable speech recogni-tion scores in quiet,and sentence recognition thresholds in quiet were assessed before fitting and on the day of 14±2 after fitting to compare differences in results.The effectiveness of the hearing aids on the day of 14±2 after fitting was also eval-uated using the IOI-HA questionnaire.Results After wearing the bone-conduction hearing aid,the average hearing thresh-old and sentence recognition threshold of the patients decreased significantly compared with before fitting(the average hear-ing threshold:56.5±8.2 dB HL before fitting,39.3±4.9 dB HL on the day of 14±2 after fitting;sentence recognition threshold:48.6±9.7 dB HL before fitting,34.3±5.6 dB HL on the day of 14±2 after fitting),and the difference was statistically significant(P＜0.001).The single-syllable speech recognition score before fitting was 29.8％±11.4％,and on the day of 14±2 after fitting,it was 72.4％±14.4％,the difference was statistically significant(P＜0.001).The av-erage total score of the IOI-HA questionnaire was 29.0±3.8 points.Conclusion Condyle-stimulated headband bone-con-duction hearing aids can significantly improve the hearing and speech recognition ability of patients with conductive,mixed and sensorineural hearing loss whose bone conduction pure tone thresholds at 0.5～4 kHz were ≤60 dB HL.It may poten-tially improve the quality of life for patients with hearing loss and holds substantial clinical application value.

Objective To study the dynamic changes in the secretion of neurotransmitters glutamic acid(Glu)and γ-aminobutyric acid(GABA)in the central auditory brain area,in order to explore the effects of sodium salicy-late on different locations of the auditory pathway.Methods A total of 126 SD rats were injected intraperitoneally with salicylate,and were divided into 10 groups including injection groups for 1,2,4,8,and 24 hours,chronic in-jection groups for 3,7,and 14 days,and chronic recovery groups for 21 and 28 days with 6 rats in each group,as well as their corresponding blank control groups.Rats in each group were anesthetized and materials were collected for further use.High-performance liquid chromatography(HPLC)was performed to detect and compare the dynam-ic changes in the levels of Glu and GABA in the auditory cortex,inferior colliculus,cochlear nucleus,and hippocam-pus of the auditory center of rats in each group at different time points.Results Compared with the control group,within 24 hours of acute injection of salicylate,the Glu content in the auditory cortex reached the peak in 1 hour,and the hippocampus reached the peak at the 4th hour after injection,and then decreased slowly.The GABA con-tent in the four brain regions showed a slow upward trend in the chronic injection period,reached the peak on the 7th day,decreased and approached normal level on the 14th day,and basically returned to the normal level in the re-covery period.Conclusion These findings indicate that salicylate has a certain short-term excitatory and stimulating effect on the central auditory system.Under the mechanism of central plasticity,after long-term injection of salicy-late,the release of neurotransmitters reaches a new excitation/inhibition balance in the central area.Glu and GABA may each play a different role that may ultimately lead to the development of tinnitus.

Objective To develop and translate the Chinese listening self-efficacy questionnaire(C-LSEQ)and to test its reliability and validity.Methods A total of 172 subjects aged≥60 years with age-related hearing loss completed the C-LSEQ questionnaire via direct interviews.Pure tone hearing threshold test,Mandarin hearing in noise test,and the hearing handicap inventory for elderly-screening(HHIE-S)were evaluated.Twenty subjects were randomly selected from the original group to complete the C-LSEQ questionnaire in 2 weeks after the initial evaluation.Results ① Reliability:The Cronbach's a coefficients of the three sub-dimensions and the overall ques-tionnaire were all＞0.8,and the test-retest reliability coefficients of the three sub-dimensions and the overall ques-tionnaire were all＞0.9(P＜0.001).② Validity:The experts collectively evaluated the representativeness of the statements with good content validity.The convergent validity test showed that the composite reliability(CR)of the 2 sub-dimensions and the overall questionnaire were all＞0.7,and the average variance extracted(AVE)were all＞0.5.The CR value of the complex listening dimension was 0.655,and the AVE value was 0.937.The criterion validity test showed that the overall questionnaire and the three sub-dimensions of the C-LSEQ were significantly correlated with the pure-tone average,speech recognition thresholds in noise,and HHIE-S(P＜0.001).Conclusion The C-LSEQ exhibits stable structure,with good reliability and validity.It can be used to assess listen-ing self-efficacy in adults with hearing loss,especially in the elderly with age-related hearing loss.

Objective To investigate the impact of the dominant deafness point mutation p.D179N on the oli-gomeric equilibrium state of Connexin 26(Cx26).Methods The wild-type Cx26 fusion protein(Cx26-WT-GFP)and mutant fusion proteins(Cx26-D179N-GFP,Cx26-D179C-GFP)were expressed in HEK293F cells.By using Fluorescence-detection size-exclusion chromatography(FSEC)and size-exclusion chromatography(SEC)to analysis the oligomeric state of the target protein based on malecular weight under the condition of solubilization and purifica-tion respectively.Cryo-electron microscopy(Cryo-EM)single particle analysis(SPA)was conducted to analysis the target protein's oligomeric states based on the 2D classification morphology of the protein particles.Results In vitro,the wild-type Cx26 protein(Cx26-WT)is almost exclusively dodecameric.The deafness mutation p.D179N protein(Cx26-D179N)exists as both dodecamers and hexamers,whereas the artificial mutation p.D179C protein(Cx26-D179C)does not form dodecamers.Conclusion The dominant deafness mutation GJB2 p.D179N could weaken the ability of docking between hexameric proteins,which could affect the balance between hexamers and do-decamers.

Objective To investigate the damage mechanism of extracellular vesicles(EVs)derived from ves-tibular schwannoma(VS)on HEI-OC1 cells and the protective effect of the ferroptosis inhibitor ferrostatin-1(Fer-1).Methods Tumor tissues and clinical data were collected from four patients with stage Ⅱ or Ⅲ VS,classified as grade D according to the AAO-HNS hearing classification.Primary VS cells were extracted,and their conditioned supernatant was collected.EVs were isolated using ultracentrifugation and identified.HEI-OC1 cells were cultured in vitro and divided into three groups:the control group(no treatment),the EVs group(treated with 3000 parti-cles/cell VS-EVs for 24 hours),and the EVs+Fer-1 group(pretreated with 20 μmol/L Fer-1 for 2 hours followed by co-culture with 3000 particles/cell VS-EVs for 24 hours).Cell viability was assessed using the CCK-8 assay,re-active oxygen species(ROS)levels were quantified using the DCFH-DA fluorescent probe,and lipid peroxidation was evaluated using the BODIPY 581/591 C11 probe.Results Compared with the control group,the EVs group showed significantly reduced cell viability(P＜0.001)and increased levels of ROS(P＜0.001)and lipid peroxides(P＜0.001).However,the EVs+Fer-1 group exhibited significantly restored cell viability(P＜0.001)and re-duced levels of ROS and lipid peroxidation(P＜0.001).Conclusion VS-derived EVs disrupts redox homeostasis,promotes intracellular accumulation of lipid peroxides and ROS,and induces ferroptosis in HEI-OC1 cells.Fer-1 significantly alleviates VS-EVs-induced ferroptosis,thereby protecting HEI-OC1 cells from damage.