Background: Mutations in the USH2A gene are strongly associated with hereditary sensorineural loss. However, the cellular mechanisms linking USH2A deficiency to cochlear hair cell vulnerability remain incompletely understood. Aims: This study aimed to investigate the effects of USH2A knockdown on Prestin expression and apoptosisrelated signaling in a cochlear hair cell model. Methods: Stable USH2A knockdown HEI-OC1 cells were generated using lentiviral shRNA transduction followed by puromycin selection (2 mu g/mL for 7 days). Gene and protein expression levels of Prestin, Bax, and Bcl-2 were assessed by RT-qPCR (2<^>(-Delta Delta Ct) method), Western blotting, and immunofluorescence. Apoptosis was evaluated by flow cytometry (Annexin V/PI), TUNEL staining, and cleaved caspase-3 immunofluorescence. Results: USH2A knockdown reduced Prestin mRNA and protein expression by approximately 60% compared with the sh-NC group (P < 0.01). Bax expression increased more than 2.5-fold, whereas Bcl-2 levels decreased by similar to 50%, indicating disruption of apoptotic balance. Flow cytometry revealed increased early apoptotic (2.16% -* 4.58%) and late apoptotic (16.3% -* 22.0%) populations. TUNEL-positive cells increased from 2.8 +/- 0.6% to 12.4 +/- 2.1% (P < 0.01), and cleaved caspase-3 fluorescence intensity was approximately threefold higher in the knockdown group. Conclusion: USH2A knockdown significantly reduces Prestin expression and promotes apoptosis through modulation of the Bax/Bcl-2 axis in HEI-OC1 cells. These findings provide mechanistic insights into how USH2A deficiency may increase cochlear hair cell vulnerability and potentially contribute to auditory dysfunction. Further in vivo validation is warranted.

Objective:Intermuscular lipoma（IL） is a rare type of deep-seated benign lipoma, accounting for approximately 0.3% of all lipomas. Its occurrence in the temporal region is particularly uncommon, and giant lesions are even rarer. We report the case of an 84-year-old male patient with a giant intermuscular lipoma in the temporal region, who had undergone two surgical excisions at an outside hospital, both followed by recurrence. The patient underwent another surgery at our hospital, during which the mass was completely excised. Postoperative histopathology confirmed the diagnosis of intermuscular lipoma, and no recurrence was observed during a 1-year follow-up. Based on the clinical presentation and treatment course of this case, combined with a review of relevant literature, the authors analyze the etiology, diagnosis and treatment, recurrence factors, and prognosis of giant intermuscular lipomas in the temporal region, aiming to improve awareness of this rare disease, emphasize intraoperative management, and reduce postoperative recurrence rates.

UNLABELLED:Hearing loss (HL) is the most common sensory disorder, with genetic factors accounting for approximately 60% of cases of congenital HL. Hearing loss is the most common sensory disorder, affecting approximately 60% of cases, which is often related to hereditary patterns. The MYO6 gene encodes the unconventional myosin VI, a protein crucial for maintaining the function of inner ear hair cells and the normal structure of stereocilia. Variants in the MYO6 have been implicated in HL at loci DFNA22 and DFNB37. Due to the high genetic heterogeneity of sensorineural HL, variants at different loci within this gene result in varying degrees of hearing impairment. This review summarizes the current understanding of MYO6-related pathogenesis, provides an overview of the structure and function of the MYO6 gene, and discusses the diseases associated with various forms of MYO6 variants. The objective is to enhance the understanding of the genotype-phenotype correlation associated with MYO6 and to explore emerging preclinical gene therapy strategies for addressing this genetically heterogeneous disorder, while also providing insights for clinical genetic counseling.

Background:Usher syndrome (USH) is an autosomal recessive disorder characterized by hearing loss, retinitis pigmentosa, and variable vestibular dysfunction. USH2A is one of the causative genes of USH. This study is aimed at exploring the mechanism of hearing loss induced by USH2A gene knockout. Method:USH2A knockout (Ush2a-/-) mice were used, and auditory brainstem response testing was performed on WT, Ush2a-/-, and Ush2a+/- mice. Then, the cochlea tissues were used to carry out immunofluorescence staining, hematoxylin and eosin (H&E) staining, and scanning electron microscopy (SEM). The mRNA expressions were detected by RT-qPCR. Finally, the differentially expressed genes (DEGs) in cochlear tissues of Ush2a-/- and WT mice were identified by transcriptome sequencing. Results:Compared to WT mice, Ush2a-/- and Ush2a+/- mice exhibited moderate-to-severe nonprogressive hearing loss, with more pronounced deficits at low (4 kHz) and high (32/24 kHz) frequencies. HE staining and immunofluorescence staining showed that the modiolus, stria vascularis, basilar membrane, and the number of inner hair cells and outer hair cells (OHCs) in USH2A knockout mice have not changed. However, SEM results showed that severe stereociliary collapse was evident in OHCs of the Ush2a-/- group. In addition, through transcriptomic analysis, 3632 upregulated genes and 2921 downregulated genes were obtained in the Ush2a-/- mice. Among these DEGs, the most DEGs associated with hearing loss were Scn2a, Shank2, Bsn, Fcer1g, Prkce, Tgfb1, and Irf7. Conclusion:This study demonstrates that USH2A deficiency disrupts auditory function through stereociliary instability and dysregulation of genes critical for synaptic transmission and cytoskeletal dynamics.

UNLABELLED:Nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs) are integral components of the cytoplasmic pattern recognition receptors (PRRs) family, playing a crucial role in both innate immunity and inflammatory responses. Nucleotide-binding oligomerization domain-like receptors detect pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs), activating multiple signaling pathways, including nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK), and triggering immune responses through inflammasome activation. The NLR family contains 5 distinguishable subfamily classifications. The development and progression of multiple ear-related disorders depend significantly on NOD1, NOD2, NLRP3, and NLRX1, among other specific members of the NLR family. The analysis investigates NLRs' interactions with ear pathologies, particularly focusing on NLRP3 functions in the development of otitis media along with its effect on cholesteatoma formation and hearing loss. In addition, this review evaluates targeted therapeutic strategies derived from NLRs research by developing a theoretical foundation that suggests new ways for advancing treatments for otological diseases.

BACKGROUND:Cochlear implantation (CI) is a standard intervention for congenital severe to profound sensorineural hearing loss. However, postoperative auditory and speech outcomes vary considerably across patients, and the developmental status of the cochlear nerve is considered a major determinant of rehabilitation efficacy. OBJECTIVE:This study aimed to assess the correlation between cochlear nerve cross-sectional area (CNCSA) and outcomes after cochlear implantation (CI). METHODS:We retrospectively analyzed 136 bilateral CI recipients, comparing auditory and speech rehabilitation results over 2 years postoperatively with bilateral CNCSA. Patients were grouped based on average CNCSA. RESULTS:A significant positive correlation was observed between CNCSA and scores on the IT-MAIS, MUSS, CAP, and SIR at 2 years post-CI. Patients with bilateral CNCSA >0.87 mm2 had significantly better speech rehabilitation outcomes than those with one or both nerves below this threshold. CONCLUSIONS:CNCSA is correlated with post-CI rehabilitation outcomes in patients with severe-to-profound sensorineural hearing loss, and may serve as a predictor for auditory and speech rehabilitation success.

IntroductionWaardenburg syndrome type 1 (WS1) is a rare autosomal dominant disorder characterized by congenital sensorineural hearing loss and facial dysmorphisms. PAX3 mutations are a known genetic cause. This study investigated a novel PAX3 mutation in a Chinese Yugur family and assessed long-term auditory outcomes after cochlear implantation.Methods and ResultsWhole-exome and Sanger sequencing were used to identify causative variants, followed by bioinformatic analyses to predict pathogenicity. Auditory and speech rehabilitation were evaluated over a 7-year follow-up. A novel heterozygous frameshift mutation in PAX3 (c.788dup, p.Gln264ThrfsTer5) was identified and confirmed to be de novo. Structural modeling indicated disruption of a conserved domain, supporting its pathogenic role. The proband achieved excellent auditory and speech recovery and successfully integrated into mainstream education.DiscussionThis study expands the mutation spectrum of PAX3 and provides evidence supporting the pathogenicity of the c.788dup variant. It also confirms the long-term benefit of cochlear implantation and rehabilitation in WS1-related hearing loss.

Presbycusis, or age-related hearing loss (ARHL), is a prevalent sensory disorder in the elderly, driven by genetic factors, oxidative stress, inflammation responses, and cellular senescence. Despite its significance, the molecular mechanisms underlying ARHL remain poorly defined. In this study, we employed bioinformatic analysis of public gene expression datasets to identify differentially expressed genes in ARHL. Protein-protein interaction network analysis further nominated LCN2 as a hub gene. Experimental validation in aging C57BL/6J mice and HEI-OC1 auditory cells revealed that elevated LCN2 expression promotes cellular senescence, while its knockdown delays this phenotype. Mechanistically, LCN2 drives senescence by activating the NF-κB signaling pathway, and its inhibition alleviates senescence induced by tert-butyl hydroperoxide (TBHP). Our findings establish LCN2 as a key pro-senescence factor in ARHL and demonstrate that it regulates auditory cell senescence through the NF-κB pathway, providing new mechanistic insights and revealing potential therapeutic targets for ARHL intervention.

Hearing loss is the most prevalent disabling disease. Cochlear implantation（CI） serves as the primary intervention for severe to profound hearing loss. This consensus systematically explores the value of genetic diagnosis in the pre-operative assessment and efficacy prognosis for CI. Drawing upon domestic and international research and clinical experience, it proposes an evidence-based medicine three-tiered prognostic classification system（Favorable, Marginal, Poor）. The consensus focuses on common hereditary non-syndromic hearing loss（such as that caused by mutations in genes like GJB2, SLC26A4, OTOF, LOXHD1） and syndromic hereditary hearing loss（such as Jervell & Lange-Nielsen syndrome and Waardenburg syndrome）, which are closely associated with congenital hearing loss, analyzing the impact of their pathological mechanisms on CI outcomes. The consensus provides recommendations based on multiple round of expert discussion and voting. It emphasizes that genetic diagnosis can optimize patient selection, predict prognosis, guide post-operative rehabilitation, offer stratified management strategies for patients with different genotypes, and advance the application of precision medicine in the field of CI.

The tectorial membrane (TM) is an essential extracellular matrix in the cochlea, integral to auditory processing by facilitating hair cell stimulation and sound transmission. Despite its vital role, the mechanisms underlying TM-related hearing loss remain unclear. This review aim to discuss the structure and functions of the TM, exploring its role in cochlear mechanics and auditory signal amplification. Abnormalities in TM composition, including disruptions in collagen, glycosaminoglycans, and non-collagenous proteins, are implicated in various forms of hearing loss, including those associated with genetic mutations and ototoxic drug exposure. We also examine the contributions of genes such as TECTA, TECTB, and CEACAM16, whose mutations disrupt TM integrity and lead to sensorineural hearing loss. Additionally, the impact of aging and thyroid hormone deficiency on TM degeneration is considered. Current diagnostic and therapeutic approaches are discussed, with an emphasis on the potential of gene therapy and stem cell therapy.

BACKGROUND:Hearing loss, a major public health issue, affects 1.33 per 1,000 live births worldwide. Genetic factors contribute to over half of congenital cases, with X-linked inheritance accounting for 1-5%. POU3F4 mutations are associated with approximately 50% of X-linked non-syndrome hearing loss cases. POU3F4 plays a critical role in cochlear development by regulating otic mesenchyme cell differentiation. The study investigates the impact of a novel POU3F4 p.E294G mutation on cochlear structure and function using cellular and animal model. METHODS:The study utilized immortalized lymphoblastoid cell lines, POU3F4 overexpressed HEK293 cells and generated Pou3f4 knock-in (Pou3f4KI) mice via CRISPR/Cas9 to introduce the p.E294G mutation. Alterations in expression and subcellular localization of POU3F4 were detected at the cellular level. Auditory function was assessed using auditory brainstem response testing. Cochlear structure was analyzed through histology, immunohistochemistry, scanning electron microscopy, and transmission electron microscopy. RNA sequencing, qPCR and Western blot were conducted to evaluate gene expression and mitochondrial function. RESULTS:The transcription of POU3F4 was abnormal and the expression was normal in lymphoblastoid cell lines. Abnormal nuclear localization of POU3F4 p.E294G was found in overexpressed HEK293 cells. Pou3f4KI mice exhibited cochlear malformations, including modiolus hypoplasia and reduced stria vascularis cell populations. Auditory testing revealed progressive hearing loss. Pou3f4 affect mitochondrial protein expression by affecting the expression of TFAM. Mitochondrial dysfunction was evident, with reduced oxidative phosphorylation (OXPHOS) complex assembly and activity, decreased ATP levels. The level of reactive oxygen species, mitochondrial fission and apoptosis in cochlea were elevated. CONCLUSIONS:The POU3F4 p.E294G resulted in abnormal nuclear localization. Pou3f4 mutant disrupts cochlear development and function, impairs mitochondrial integrity, induces oxidative stress, and promotes apoptosis, leading to progressive hearing loss. The findings enhance the understanding of POU3F4-related hearing loss mechanisms and highlight the importance of early genetic screening and audiological monitoring.

ABSTRACTBackgroundMYO15A is one of the common genes of severe‐to‐profound sensorineural deafness. Mutations in this gene can cause both pre‐ and post‐lingual hearing losses. In this study, a novel MYO15A variant (c.2482C>T) was identified to be associated with autosomal recessive non‐syndromic hearing loss (ARNSHL) in a Chinese Uighur family.MethodsTo examine the effects of the MYO15A mutation on the morphology and function of the derived hair cell‐like cells, two iPSCs were generated separately from the proband and a mutation‐negative family member and those were then induced to hair cell‐like cells.ResultsResults showed that this homozygous MYO15A mutation (PVS1 + PM2 + PP1 + PP3), which is located in the N‐terminal domain, displayed significant differences in the morphology and function of hair cell‐like cells between the proband and the normal control, although it had no effect on the totipotency of iPSCs.ConclusionOur study demonstrates that the novel variant c.2482C>T in the MYO15A gene may cause inner ear hair cell dysfunction and audiological disorders in this family.

Large vestibular aqueduct syndrome（LVAS） is a common recessive hereditary hearing loss disease, and some patients may also experience vestibular dysfunction. With the wide application of cochlear implant（CI） and the development of vestibular medicine, the pathophysiological mechanism of LVAS and the influence mechanism of CI on vestibular function are gradually elucidated. Consequently, the evaluation and rehabilitation of vestibular dysfunction function have also become research hotspots. This article reviews studies on vestibular function and related rehabilitation in patients with large vestibular aqueduct syndrome.

Retiform hemangioendothelioma (RH) is a rare intermediate (locally aggressive) vascular tumor that mostly affects the dermis of the trunk and limbs, but has never been reported in the inferior turbinate. A 10-year-old Chinese boy presented with recurrent epistaxis in his left nasal cavity and anemia for more than 2 years. Radiographic and electronic video laryngoscopic images showed an expansile mass in the left inferior turbinate. Endoscopic surgery and electrocautery were performed to resect the tumor beyond the macroscopic border. Histopathologically, the tissues were infiltrated by hyperplastic blood vessels arranged in a retiform pattern, and endothelial cells proliferate significantly in some areas. Immunohistochemistry showed a positive result for CD31, CD34, Fli-1, and ERG. No epistaxis, tumor recurrence, or metastasis was found on reexamination over 18 months after surgery.

Background:Aminoacyl-tRNA synthetases are highly conserved proteins that catalyze the tRNA aminoacylation reaction to produce aminoacyl-tRNAs involved in protein synthesis, which are required to translate cytoplasmic and mitochondrial proteins. The mt-ARS genes encode the mitochondrial aminoacyl-tRNA synthetase (mt-ARSs), and variants in mt-ARS genes affect mitochondrial protein synthesis. This can impair the translation of mitochondrial proteins, adversely affecting oxidative phosphorylation and leading to related diseases. To date, 19 mt-ARS genes have been identified and found to be strongly associated with the development of mitochondrial disorders. Hearing loss (HL) is one of the most common chronic conditions in children and a leading cause of communication disorders. Genetic studies of sensorineural HL are critical to diagnosing and treating sensorineural HL. The relationship between mt-ARS genes and sensorineural HL is gradually surfacing as cases of HL phenotypes caused by variants in the mammalian mt-ARS genes continue to be reported. Seven mt-ARS genes have been reported to contribute to various hereditary sensorineural HL. Summary:This article reviews studies on mitochondrial aminoacyl-tRNA synthetase, mt-ARS genes, and variants associated with HL phenotypes. Investigating their genetic characteristics provides deeper insights into the pathophysiology and molecular mechanisms of sensorineural hearing loss. Key Messages:Disease phenotypes resulting from variants in mt-ARS genes exhibit significant clinical heterogeneity. The varying degrees of sensorineural HL phenotypes caused by mt-ARS gene variants warrant the attention of otologists and researchers. At least seven of the currently reported mt-ARS genes are associated with sensorineural HL. However, the molecular mechanisms by which these genes contribute to HL remain incompletely understood. Further studies of the mt-ARS genes still await additional case reports, as well as related model animal studies and combined functional studies.

Defects in mitochondrial RNA metabolism have been linked to sensorineural deafness that often occurs as a consequence of damaged or deficient inner ear hair cells. In this report, we investigated the molecular mechanism underlying a deafness- associated tRNA(Phe) 593T > C mutation that changed a highly conserved uracil to cytosine at position 17 of the DHU-loop. The m.593T > C mutation altered tRNA(Phe) structure and function, including increased melting temperature, resistance to S1 nuclease-mediated digestion, and conformational changes. The aberrant tRNA metabolism impaired mitochondrial translation, which was especially pronounced by decreases in levels of ND1, ND5, CYTB, CO1, and CO3 harboring higher numbers of phenylalanine. These alterations resulted in aberrant assembly, instability, and reduced activities of respiratory chain enzyme complexes I, III, IV, and intact supercomplexes overall. Furthermore, we found that the m.593T > C mutation caused markedly diminished membrane potential, and increased the production of reactive oxygen species in the mutant cell lines carrying the m.593T > C mutation. These mitochondrial dysfunctions led to the mitochondrial dynamic imbalance via increasing fi ssion with abnormal mitochondrial morphology. Excessive fi ssion impaired the process of autophagy including the initiation phase, formation, and maturation of the autophagosome. In particular, the m.593T > C mutation upregulated the PARKIN-dependent mitophagy pathway. These alterations promoted an intrinsic apoptotic process for the removal of damaged cells. Our fi ndings provide critical insights into the pathophysiology of maternally inherited deafness arising from tRNA mutation-induced defects in mitochondrial and cellular integrity.

Objective: To investigate the consistency between the hearing handicap inventory (HHI) and pure-tone audiometry (PTA) scores in assessing hearing status to provide valuable insights for clinical application. Methods: Retrospective analysis of clinical data and the HHI reporting status of 6540 patients admitted between April 2020 and July 2022 for self-reported unilateral hearing loss who met the study inclusion and exclusion criteria. The kappa coefficient was used to evaluate the consistency of HHI and PTA in assessing the hearing status of the participants. Results: The PTA results showed that among the 6540 participants, 3895 exhibited normal hearing, 1434 showed mild hearing loss, 809 presented with moderate hearing loss, and 402 showed severe hearing loss. The mean hearing thresholds from 0.5 to 4 kHz in healthy ears ranged from 3.65 to 18.45 dB HL, with a mean of 10.83 +/- 5.29 dB HL; in ears affected by hearing loss, this ranged from 35 to 125 dB HL, with a mean of 69.63 +/- 28.45 dB HL. The HHI scores showed that 4820 people had normal hearing, 1245 had mild-to-moderate hearing loss, and 475 had severe hearing loss. The kappa coefficients of normal, mild-to-moderate, and severe hearing loss were 0.312, 0.223, and 0.716, respectively (P = .001). The consistency between the 2 groups was particularly significant in the assessment of severe hearing loss. Using the PTA results as a benchmark, the sensitivity, specificity, positive predictive value, and negative predictive value of the HHI were found to be 73.08%, 87.83%, 95.60%, and 70.98%, respectively. Conclusion: The HHI and PTA results were consistent in the assessment of hearing status, particularly in the assessment of severe hearing loss, and the level of consistency between the 2 methods was high. The combined use of these tools can facilitate a comprehensive assessment of the auditory status of patients with hearing loss.

Abstract Background Usher syndrome type 3 (USH3) is an autosomal recessive inherited disorder caused by pathogenic variants in the CLRN1 gene. Object To evaluate the genotype-phenotype correlation of Usher syndrome type 3 (USH3) in a deaf-blind Chinese family of 3 generations with 2 patients. Methods We collected blood samples and clinical data from all of the pedigree family members. Genomic DNA was isolated from peripheral leukocytes using standard method. Targeted next generation sequencing and Sanger sequencing were performed to find the pathogenic variants in this family. Digital PCR and plasmid overexpression assay were used to verify the pathogenicity of variant sites in different transcripts. Results All patients developed bilateral sensorineural hearing loss (SHL), progressive vision loss and nyctalopia. NGS of genes for Usher syndrome, deafness and retinal dystrophy identified a locus mutation in CLRN1 that caused completely different amino acid changes in different transcripts[CLRN1:c.474T > A(P.Cys158Ter) at NM_001256819.2 or c.302T > A(p.Val101Asp) at NM_174878.3], and plasmid overexpression experiments confirmed that the c.474T > A(P.Cys158Ter, NM_001256819.2) was a pathogenic variant which has never been associated with Usher syndrome in China, and the transcript of this mutation was not the version commonly found worldwide. Conclusions The CLRN1c.474T > A(NM_001256819.2) mutation is the causative variant in the Chinese family with USH3. The pathogenicity of different transcripts should be particularly considered in pathogenicity analysis.

BackgroundCochlear implants (CI) help regain perception of sound for patients with sensorineural hearing loss. The ability to recognize music pitch may be crucial for recognizing and producing speech for Mandarin. Aims/ObjectivesThis study aims to search for possible influencing factors of music perception and correlations between music perception and auditory speech abilities among prelingually deaf pediatric Mandarin-speaking CI users. Material and MethodsMusic perception of 24 pediatric CI users and 12 normal hearing children was measured using the MuSIC test. Auditory speech perception of the 24 CI users was also measured and analyzed with their music perception results. ResultsPediatric CI users performed worse than normal hearing children in pitch, rhythm and melody discrimination tests (p < .05). Significant difference in pitch and melody discrimination tests between age at implantation <5 and >5 groups was found. There were significant correlations between perception of consonants, tones, and speech in a noisy environment and perception of music pitch and melody. Conclusion and SignificancePrelingually deaf pediatric CI users who received implantation before the age of five perform better in music perception tests. Pediatric CI users with better music perception show better auditory speech perception of Mandarin.