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.

The pathogenesis of tinnitus is unclear and may involve the peripheral and central nervous sys-tems.Recent studies have found that neuroinflammatory response leads to hyperactivity of nerve center,in which tumor necrosis factor-α(TNF-α)has an important impact on neurotransmitter transmission,transmembrane ion concentration,neuronal synaptic structure and functional reorganisation,and may play an important role in the path-ogenesis of acute tinnitus.This article discusses the central mechanism of TNF-α in mediating the onset of acute tin-nitus.

Objective To study the impact of tympanic membrane opening on respiratory-driven middle ear pressure in patients with patulous eustachian tube(PET),using a simplified in vitro model.Methods CT imaging data from a PET patient(with full-length eustachian tube opening observed during a Valsalva maneuver followed by breath-holding)were used to design a simplified eustachian tube model.Two simplified in vitro models of the eusta-chian tube were constructed using silicone-based 3D printing technology and connected to a pressure controller and pressure sensors.The pressure controller was activated to introduce negative-pressure airflow into the nasopharyn-geal model to simulate respiratory-induced middle ear pressure fluctuations.A hemostat was used to alternately open and close the external interface of the middle ear chamber,simulating conditions of an open and intact tympanic membrane,while middle ear pressure was continuously monitored using pressure sensors.Results In the first mod-el,with-800 mbar negative pressure applied at the nasopharynx,the middle ear pressure stabilized between-3.9 mbar and-4.3 mbar with tympanic membrane opening,and between-7.9 mbar and-8.2 mbar with intact tym-panic membrane.In the second model,under the same pressure setting,middle ear pressure stabilized between-2.7 mbar and-3.1 mbar with tympanic membrane opening,and between-5.0 mbar and-7.7 mbar with intact tympanic membrane.Conclusion This study,based on a simplified in vitro model,demonstrates that tympanic membrane opening can effectively reduce respiratory-driven pressure in the middle ear.This phenomenon may partly explain the clinical efficacy of tympanostomy tube insertion in certain PET patients.

Objectiv To investigate an autosomal dominant non-syndromic hearing loss family pedigree com-prehensively,aiming to precisely define its clinical phenotypes and uncover the underlying molecular genetic etiolo-gy.Methods A detailed interrogation of the proband's medical history and family history was conducted.Physical examinations,audiological assessments and temporal bone CT scans were performed.Genomic DNA was extracted from the peripheral blood of the proband(Ⅳ-8)for whole-exome sequencing(WES).Subsequently,candidate vari-ants identified through WES were validated among family members using Sanger sequencing.Results There were 36 individuals in 4 generations in this family pedigree,showing autosomal dominant inheritance.Among them,16 individuals presented with progressive hearing loss.Audiological examinations were completed for 13 of them,re-vealing normal hearing in three individuals(Ⅲ-1,Ⅲ-1 1,Ⅳ-4)and bilateral symmetric hearing loss of varying se-verity in the remaining ten(Ⅱ-2,Ⅱ-4,Ⅱ-10,Ⅲ-4,Ⅲ-10,Ⅲ-13,Ⅲ-14,Ⅳ-1,Ⅳ-7,Ⅳ-8),and the degree of hearing loss was related to age.WES of Ⅳ-8 revealed that she carried the variant NM_199330.2(HOMER2):c.1064 A＞G(p.Ter354Trpext10),and Sanger sequencing verified the variation at this site.Peripheral blood samples of 18 individuals in this family were collected in total.All affected individuals(Ⅱ-2,Ⅱ-4,Ⅱ-10,Ⅲ-4,Ⅲ-9,Ⅲ-10,Ⅲ-13,Ⅲ-14,Ⅳ-1,Ⅳ-7,Ⅳ-8)carried the HOMER2 c.1064 A＞G variant,except for one young member(Ⅳ-6)who had not yet developed hearing loss.Unaffected individuals(Ⅱ-5,Ⅲ-1,Ⅲ-5,Ⅲ-11,Ⅳ-2,Ⅳ-4)lacked the variant,demonstrating complete cosegregation of genotype and phenotype.According to ACMG guide-lines,this variant was classified as likely pathogenic(PM2+PP1+PM4).Conclusion The c.1064 A＞G(p.Ter354Trpext10)variant of the HOMER2 gene is the molecular genetic etiology of this hereditary deafness family pedigree.

Objective To study the clinical efficacy of buccal acupuncture in the treatment of refractory tinnitus in the elderly.Methods In this study,50 elderly patients with refractory tinnitus were randomly divided into two groups:the experimental group(n=26)received buccal acupuncture therapy on the neck,upper neck,scapula zone,shoulder,back,mastoid process,thoracic plexus,abdominal plexus and pelvic plexus on the affected side,and the control group(n=24)received sound therapy.After 8 weeks of treatment intervention,the two groups were comprehensively evaluated for the changes in the tinnitus evaluation scale(TEQ),self-rating depression scale(SDS),and blood rheological indexes,including whole blood high-cut viscosity(HSV),plasma viscosity(PSV)and fibrinogen in blood(FIB)clotting.Results After treatment,the total effective rate of the experimental group was 84.6％(22/26),higher than 54.2％(13/24)in the con-trol group.The difference in total effective rate and efficacy level between the two groups was statistically significant(P＜0.05).TEQ and SDS were significantly lower in the two groups compared to pre-treatment(P＜0.05),and HSV,PSV and FIB in the experimental group were significantly lower compared to pre-treatment(P＜0.05).The experimental group demonstrated better post-treatment outcomes in TEQ,SDS,HSV,and FIB compared to the control group(P＜0.05).Conclusion Buccal acupuncture treatment is effective in improving the symptoms of refractory tinnitus in the elderly,relie-ving the depression complicated by tinnitus,and is helpful in changing blood flow resistance and reducing blood coagulation probably.

Objective To investigate the role of the prefrontal cortex in the generation and adaptation of chro-nic subjective tinnitus using resting-state functional magnetic resonance imaging(rs-fMRI).Methods Resting-state functional magnetic resonance imaging scan were acquired from 20 patients with chronic subjective tinnitus and 20 healthy controls.Fractional amplitude of low-frequency fluctuations(fALFF)and seed-based whole-brain functional connectivity(FC)methods were used to detect abnormal prefrontal cortex activity in tinnitus patients and to investi-gate interactions between prefrontal cortex activity and brain regions associated tinnitus perception.The analysis aimed to assess the relationship between prefrontal cortex spontaneous neural activity,atypical functional connectivi-ty across various brain regions,and clinical characteristics of tinnitus.Results Compared with healthy controls,pa-tients with chronic tinnitus showed a significant reduction in fALFF values in some specific brain areas of prefrontal cortex,including the left/right medial superior frontal gyrus and the left/right middle frontal gyrus.Functional con-nectivity values were notably enhanced between the left medial superior frontal gyrus and the left anterior insula,as well as between the right medial superior frontal gyrus and the left superior temporal gyrus.Furthermore,increased functional connectivity was observed between the left middle frontal gyrus and the right middle temporal gyrus,as well as between the right middle frontal gyrus and the left parahippocampal gyrus,left superior parietal lobule,and left supplementary motor area.Importantly,the functional connectivity between the left middle frontal gyrus and the right superior temporal gyrus exhibited a negative correlation with tinnitus handicap inventory scores(r=-0.627,P=0.003)and visual analogue scale scores(r=-0.596,P=0.005).Conclusion There are abnormal brain function changes in medial prefrontal cortex and dorsolateral prefrontal cortex in patients with chronic subjec-tive tinnitus,accompanied by changes in the intensity of functional connections with the salience and auditory net-works.These abnormalities are highly related to the severity of tinnitus.The prefrontal cortex may play an impor-tant role in the sensory prediction and auditory regulation of tinnitus.

Objective To establish normal reference values for the sensory organization test(SOT)for flight crews of different ages from 20 to 50 years old,and to investigate the effect of age on the results of the SOT.Meth-ods A total of 122 flight crew members were divided into three groups of 20～29(n=57),30～39(n=44),and 40～50 years old(n=21).The EquiTest Dynamic Balance System was applied to test the SOT of 122 flight crew members.Equilibrium score,composite equilibrium score,the sensory weighting score of somatosensory,visual,vestibular,and visual preference,the strategy analysis scores,and the center of gravity alignment under six condi-tions were analyzed.Results The equilibrium scores for conditions 1 to 6 were 94.41±1.47,93.27±1.96,92.57±2.74,85.01±5.43,70.62±7.23,and 69.08±7.84,respectively,and the composite equilibrium score was 81.31±3.38.The sensory weighting scores of somatosensory,visual,vestibular,and visual preference were 98.97±2.08,89.57±5.14,73.89±7.95,and 98.84±5.78,respectively.The strategy analysis scores for conditions 1 to 6 were 95.57±1.23,94.87±1.58,93.93±5.12,88.21±4.01,79.49±7.90,and 80.55±6.57,respectively.There were significant differences in equilibrium score and somatosensory sensory weighting scores for condition 2 across age groups in this study,and the differences in other parameters between age groups were not statistically significant.Conclusion This study establishes normal reference values of SOT for flight crews of different ages to provide an objective reference for the selection of flight cadets and the medical evaluation of flight crews'sensory organization ability of balance function.

Objective To compile a comprehension assessment scale of double-conditional phrases for adults and to test its reliability and validity.Methods With reference to the compilation process of local and international word lists,a comprehension assessment scale of double-conditional phrases for adults was developed.The scale was used to assess 40 aphasia patients and 20 ordinary adults as subjects from four hospitals in Shanghai.The internal consistency reliability,split-half reliability,retest reliability and calibration validity of the scale were evaluated and analyzed[the western aphasia battery(WAB)as a calibrated measurcment instrument].Results The scale consists of 30 questions,including five types of phrases:coordinate phrases,verb-object phrases,subject-predicate phrases,endocentric phrases and interface-object phrases.Compared with the ordinary adult group,the total score of the scale and the scores of each dimension in the aphasia group were significantly lower than those in the control group(P＜0.001).The Cronbach's alpha coefficient for the total score was 0.894,the half coefficient was 0.924,and the correlation coefficient for the retest reliability of the total score was 0.923,with retest reliability for each type of word group ranging from 0.731 to 0.903.With the exception of the repetition scores,correlation coefficients be-tween the scale and all other scores of WAB were all＞0.6(P＜0.01).Conclusion The self-developed"Adult Double Conditional Phrases Auditory Comprehension Assessment Scale"demonstrates good reliability and validity.It serves as a scientific and reliable tool for evaluating the auditory comprehension ability of adult patients.

Objective To investigate the expression and significance of RANK/RANKL/OPG signaling path-way in gerbil external auditory canal cholesteatoma(EACC).Methods Twenty-five mongolian long-clawed gerbils were divided into an experimental group(n=15)and a control group(n=10),and an EACC model was constructed by external auditory canal ligation method in the experimental group.Three months later,the external auditory ca-nal tissues of the control group and the experimental group(10 ears)were collected,and RT-qPCR and Western blot were performed to detect the expression levels of receptor activator of NF-κB(RANK),receptor activator of NF-κB ligand(RANKL),osteoprotegerin(OPG)mRNA and protein in the signaling pathway.Results Two ger-bils in the experimental group died during the modeling process,and cholesteatoma of the external auditory canal was formed in 10 ears 3 months after the operation,with a modeling success rate of 76.9％(10/13).Compared with the normal control group,there was no statistically significant difference in the expression of RANK mRNA in gerbil external auditory canal tissues(P=0.08).The expression of RANK protein was upregulated,the expression of RANKL mRNA and protein was upregulated,while the expression of OPG mRNA and protein was downregulat-ed,the ratio of RANKL/OPG was upregulated,with a statistically significant difference between the two groups(P＜0.01).Conclusion The RANK/RANKL/OPG signaling system may play an important role in the formation of gerbil EACC and bone destruction.

Alzheimer's disease(AD)is an age-related neurodegenerative disease with an increasing incidence worldwide.A large number of studies have shown that the incidence rates of hearing loss is high in patients with mild cognitive impairment and Alzheimer's disease,and may be a risk factor for the occurrence and development of cognitive impairment.There is an interaction between the two,but the causal mechanism is still unclear.Early screening and management of hearing impairment may play an important role in the early diagnosis,symptom im-provement and disease progression of Alzheimer's disease.This paper reviews relevant clinical and basic research to discuss the correlation between hearing loss and Alzheimer's disease,and the possible causal mechanism between them.