Ferroptosis is a recently discovered iron-dependent form of oxidative programmed cell death distinct from caspase-dependent apoptosis. In this study, we investigated the effect of ferroptosis in neomycin-induced hair cell loss by using selective ferroptosis inhibitor liproxstatin-1 (Lip-1). Cell viability was identified by CCK8 assay. The levels of reactive oxygen species (ROS) were determined by DCFH-DA and cellROX green staining. The mitochondrial membrane potential () was evaluated by TMRM staining. Intracellular iron and lipid peroxides were detected with Mito-FerroGreen and Liperfluo probes. We found that ferroptosis can be induced in both HEI-OC1 cells and neonatal mouse cochlear explants, as evidenced by Mito-FerroGreen and Liperfluo staining. Further experiments showed that pretreatment with Lip-1 significantly alleviated neomycin-induced increased ROS generation and disruption in in the HEI-OC1 cells. In parallel, Lip-1 significantly attenuated neomycin-induced hair cell damage in neonatal mouse cochlear explants. Collectively, these results suggest a novel mechanism for neomycin-induced ototoxicity and suggest that ferroptosis inhibition may be a new clinical intervention to prevent hearing loss.

Our objective was to study the characteristics of patients with subjective tinnitus and normal hearing and to investigate whether the features correlated to different shapes on audiograms. In this retrospective study, 313 patients with subjective tinnitus and clinically normal hearing were enrolled from the tinnitus outpatient department of the Eye and ENT Hospital of Fudan University. The following phenotypic variables were collected: age, dominant tinnitus pitch (TP), tinnitus loudness, tinnitus duration, tinnitus severity, sex, education, hearing thresholds, tinnitus position, and tinnitus condition. The dominant TPs of patients with normal hearing were mostly high-pitched, with a mean of 4866.8 ± 2579.6 Hz; thus, we speculated that the condition is related to high-frequency hearing threshold elevations. We further divided the patients into four subgroups based on the matched TP: (i) TP ≤ 500 Hz (n = 34), (ii) 500 Hz < TP ≤ 3,000 Hz (n = 15), (iii) 3,000 Hz < TP ≤ 8,000 Hz (n = 259), and (iv) TP > 8,000 Hz (n = 5). We studied the phenotypic profiling of different audiograms and found that the group with TP of ≤500 Hz had an average “inverted-U” shaped audiogram, and the group with TP between 500 and 3,000 Hz had a slowly ascending slope audiogram below 2,000 Hz, followed by a drastically descending slope audiogram ranging from 2,000 to 8,000 Hz; further, the high-frequency (3,000–8,000 Hz) and ultra-high-frequency (>8,000 Hz) groups had flat curves below 2,000 Hz and steeper slope audiograms over 2,000 Hz. Our findings confirmed a consistency ratio between the distributions of dominant TPs and the frequencies of maximum hearing thresholds in both ears. The dominant TP was positively correlated with the maximum hearing threshold elevation frequency (left ear: r = 0.277, p < 0.05; right ear: r = 0.367, p < 0.001). Hearing threshold elevations, especially in high frequency, might explain the appearance of dominant high-frequency TP in patients without clinically defined hearing loss. This is consistent with the causal role of high-frequency coding in the generation of tinnitus.

Objectives: To explore the role of DNA methyltransferase 1 (DNMT1) in the development of auditory system using zebrafish as experimental model.Methods: Morpholino oligonucleotide was used to induce Dnmt1 deficiency. RNA sequencing, in situ hybridization (ISH), whole genomic bisulfide sequencing (WGBS) and immunostaining were used to investigate the morphologic alterations and mechanisms.Results: We found that downregulation of Dnmt1 induced decreased number of neuromasts and repressed cell proliferation of primordium in the developing posterior lateral line system of zebrafish. The ISH data uncovered that Fgf signalling pathway was inhibited and the expression of chemokine memberscxcr4b,cxcr7bandcxcl12awere interfered, whilelef1expression was increased after inhibiting Dnmt1. Additionally, Dnmt1 downregulation led to malformed otoliths and deformed semicircular canals, and hair cell differentiation in utricle and saccule was inhibited severely. The in situ staining of otic placode markerspax2/5andfgf 3/8/10was decreased when Dnmt1 downregulated. The WGBS analysis demonstrated that the global methylation status was markedly downregulated, and cell cycle genes were among those most differently expressed between Dnmt1 morphants and the controls. Further ISH analysis confirmed the findings by RNA‐seq and WGBS assay thatcdkn1aandtp53were both upregulated after knockdown of Dnmt1.Conclusion: Our results revealed that Dnmt1 is essential for the development of zebrafish auditory organ through regulating cell cycle genes together with Wnt and Fgf signalling pathways.

Introduction Tinnitus is a common ailment that affects 10%–15% of adults worldwide. Comorbidities associated with tinnitus often include hearing loss, emotional distress and sleep disorders, with insomnia being a common issue among patients with tinnitus. Audiological and psychological approaches are typically used to treat chronic subjective tinnitus, with the combination of sound therapy and cognitive–behavioural therapy-based counselling having the strongest evidence for effectiveness. Acceptance and commitment therapy (ACT) has also shown promising effects in reducing the impact of tinnitus when delivered in groups or online. However, there is a lack of evidence on the effects of combining ACT with sound therapy for patients with tinnitus-related insomnia. Therefore, this study aims to compare the efficacy of internet-delivered ACT combined with sound therapy versus sound therapy alone for tinnitus in a superiority, two-arm randomised controlled trial. Methods and analysis A total of 164 patients with chronic subjective tinnitus and insomnia will be randomised to receive internet-delivered guided self-help tinnitus treatment based on ACT combined with tailored sound therapy or tailored sound therapy alone. The primary outcome is the variation in Tinnitus Handicap Inventory scores observed 2 months after randomisation between the two study groups. Secondary outcomes will include insomnia severity, sleep parameters, tinnitus loudness, tinnitus acceptance, depression and anxiety. The outcomes will be assessed at 2, 3 and 6 months post randomisation. Ethics and dissemination The study is approved by the review board and ethics committee of the Eye and ENT Hospital of Fudan University (approval number: 2023066-1). The findings will be disseminated through presentations at relevant conferences and peer-reviewed publications. Trial registration number NCT05963542.

Background: Tinnitus is a major health issue, but currently no tinnitus elimination treatments exist for chronic subjective tinnitus. Acoustic therapy, especially personalized acoustic therapy, plays an increasingly important role in tinnitus treatment. With the application of smartphones, personalized acoustic stimulation combined with smartphone apps will be more conducive to the individualized treatment and management of patients with tinnitus. Objective: The aim of this study was to evaluate the efficacy of a new personalized approach known as the digital frequency customized relieving sound (DFCRS) for tinnitus treatment and to explore the factors that may influence its therapeutic effect. Methods: Patients with subjective tinnitus were enrolled in this study from July 14, 2020, to May 24, 2021, in the tinnitus specialist clinic of Eye and ENT Hospital, Fudan University, Shanghai, China. In this nonrandomized concurrent controlled trial, a total of 107 participants were assigned to listen to personalized DFCRS through our developed app, while the other 77 participants who did not want to download and use the app were assigned to listen to unmodified music (UM). All the recruits were instructed to listen to DFCRS or UM for at least 2 hours a day and complete follow-up assessments at baseline, 1, 2, and 3 months. Multidimensional assessment scales, that is, Tinnitus Handicapped Inventory (THI), Hospital Anxiety and Depression Scale (HADS), Athens Insomnia Scale (AIS), Fear of Tinnitus Questionnaire (FTQ), and Tinnitus Catastrophizing Scale (TCS) were used to evaluate the severity of tinnitus and the quality of life. Linear mixed models were used to test for changes in the THI scores across 3 months of acoustic treatment between group (DFCRS or UM treatment) and time. A multiclass logistic model was built with a stepwise function to determine the influence of the different covariates on the effects of acoustic treatment. Results: The results of the multidimensional assessment scales after 3 months of treatment showed that DFCRS-treated patients had significant tinnitus relief compared to those in the UM group. Linear mixed models revealed a significant reduction in the THI scores over time (P<.001), with the DFCRS group showing significantly greater improvement than the UM group (P<.001). At 3 months, 92.5% (99/107) of the patients undergoing DFCRS reported tinnitus relief or disappearance, and longer daily treatment time was associated with better outcomes (P=.007). Multiclass logistic regression confirmed that longer treatment time (odds ratio [OR] 13.07-64.78; P<.001) and more severe tinnitus at baseline (OR 10.46-83.71; P<.001) predicted better treatment response. All secondary outcomes (HADS, AIS, FTQ, TCS) showed significant improvements over time (P<.001). Conclusions: Our study suggests that DFCRS is a new promising and noninvasive therapy for chronic tinnitus, and it can be delivered through a mobile app to bring more convenience to patients with tinnitus.