Background/Objectives: Branchio-oto syndrome (BOS) is an autosomal dominant disorder primarily associated with pathogenic variants in EYA1, mainly characterized by branchial anomalies, auricular abnormalities, and hearing loss. However, the co-occurrence of inner ear malformations in BOS remains understudied, especially severe malformations. This study aimed to investigate the clinical and genetic characteristics of patients with EYA1-associated BOS, with emphasis on cochlear nerve deficiency (CND). Methods: From January 2020 to April 2026, patients diagnosed with EYA1-associated BOS at an otology outpatient clinic in a tertiary hospital were included. Clinical manifestations, audiological assessments, imaging and genetic findings were analyzed. Results: Thirteen patients (six females and seven males) from eight unrelated families aged 0.3–58.3 years were enrolled. Branchial cleft fistulas and preauricular pits were each observed in 76.9% (10/13) of patients. The mean pure-tone average was 74.3 ± 20.7 dB HL. Eight EYA1 variants (four truncating, two large deletions, and two splicing) were identified. Among these, five were novel (c.320_329del, c.518del, c.1307dupT, c.1475+1G>A, and exon 12–18 deletion). CND was detected in 38.5% (5/13) of patients and 26.9% (7/26) of ears. Patients with CND carried either truncating variants (n = 3) or large deletions (n = 2) of EYA1. No CND was observed in patients with splicing variants. Conclusions: This study identifies five novel EYA1 pathogenic variants and suggests that CND may be a relatively common radiologic feature in EYA1-associated BOS, particularly among patients with truncating variants or large deletions, although larger studies are needed to confirm this association.

Gemcitabine (GEM) is a cornerstone of chemotherapy for lung adenocarcinoma (LUAD). However, acquired resistance severely limits its clinical efficacy, leading to relapse and metastasis. This study aims to investigate the role of KNSTRN in GEM resistance and its underlying molecular mechanisms in LUAD. Bioinformatics analysis revealed the expression levels of KNSTRN in LUAD tissues and the enriched pathways, and analyzed the prognostic impact of KNSTRN in LUAD, predicting the upstream transcription factor (TF) ATF2. The study examined the correlation and binding sites between KNSTRN and ATF2, as well as the expression of ATF2 in LUAD tissues. Molecular and cellular experiments were conducted to depict the binding relationship between ATF2 and KNSTRN and their expression within cells; to assess cell viability and IC50 values; to evaluate the expression of proteins related to the mismatch repair pathway, glycolytic pathway, and DNA damage markers; and to measure extracellular acidification rate, oxygen consumption rate, lactate production, and glucose consumption. Bioinformatics evidence pointed to ATF2 being the upstream TF for KNSTRN, with their expressions positively linked. High levels of ATF2 and KNSTRN were detected in LUAD tissues and cells, and high KNSTRN expression was associated with poor prognosis. KNSTRN promoted glycolysis and lactate accumulation, which in turn suppressed the expression of key MMR components MLH1 and MSH2, thereby impairing DNA damage repair capacity correlated positively with the expression of pivotal genes in these pathways. Molecular and cellular studies corroborated that KNSTRN overexpression enhances GEM resistance, a process that is thought to be mediated by increased lactate production and the regulation of mismatch repair pathways. Rescue experiments showed that the overexpression of KNSTRN reversed the effects of ATF2 silencing on LUAD cell GEM resistance. This study has uncovered a novel mechanism by which ATF2 activates KNSTRN, leading to increased lactate production and the downregulation of MLH1 and MSH2 expression, key components of the mismatch repair machinery, thereby promoting GEM resistance in LUAD. This discovery offers new insights for improving the efficacy of chemotherapy in LUAD.

Universal newborn hearing screening (UNHS) is recognized as the most effective strategy for early detection of congenital hearing loss; however, screening coverage remains inadequate in many countries. In China, newborn hearing screening has been implemented for over two decades. To evaluate our policies and practices during this period and assess resource equity, health impacts, and future challenges, we conducted a nationwide survey focusing on newborn hearing screening coverage, the number of special schools for deaf-mutes, and the proportion of hearing-impaired students in mainstream education. From 2001 to 2020, China's UNHS program coverage increased from 10.9% to 94.3%, while the proportion of hearing-impaired students in mainstream education rose from 24.8% to 57.5%. Concurrently, the number of hearing-impaired students in special schools decreased from 76,554 to 34,945, and the number of special schools for deaf-mutes declined from 639 to 389. Through the implementation of the UNHS program, China has made substantial progress in improving newborn hearing health, yielding long-term benefits for those with congenital hearing loss. However, targeted resource allocation and the establishment of a national platform remain priorities for future development. Our experience may provide valuable insights for similar settings.

Intraoperative cell salvage (IOCS) has been widely applied as an important blood conservation measure in surgical operations. However, there is currently a lack of clinical practice guidelines for the implementation of IOCS in patients with malignant tumors. This report aims to provide clinicians with recommendations on the use of IOCS in patients with malignant tumors based on the review and assessment of the existed evidence. Data were derived from databases such as PubMed, Embase, the Cochrane Library and Wanfang. The guideline development team formulated recommendations based on the quality of evidence, balance of benefits and harms, patient preferences, and health economic assessments. This study constructed seven major clinical questions. The main conclusions of this guideline are as follows: 1) Compared with no perioperative allogeneic blood transfusion (NPABT), perioperative allogeneic blood transfusion (PABT) leads to a more unfavorable prognosis in cancer patients (Recommended); 2) Compared with the transfusion of allogeneic blood or no transfusion, IOCS does not lead to a more unfavorable prognosis in cancer patients (Recommended); 3) The implementation of IOCS in cancer patients is economically feasible (Recommended); 4) Leukocyte depletion filters (LDF) should be used when implementing IOCS in cancer patients (Strongly Recommended); 5) Irradiation treatment of autologous blood to be reinfused can be used when implementing IOCS in cancer patients (Recommended); 6) A careful assessment of the condition of cancer patients (meeting indications and excluding contraindications) should be conducted before implementing IOCS (Strongly Recommended); 7) Informed consent from cancer patients should be obtained when implementing IOCS, with a thorough pre-assessment of the patient's condition and the likelihood of blood loss, adherence to standardized internally audited management procedures, meeting corresponding conditions, and obtaining corresponding qualifications (Recommended). In brief, current evidence indicates that IOCS can be implemented for some malignant tumor patients who need allogeneic blood transfusion after physician full evaluation, and LDF or irradiation should be used during the implementation process.

Objective:Prostate cancer (PCa) often metastasizes to the bone, posing a significant clinical challenge. This study aims to develop a bone metastasis-related risk model for PCa. Methods:Bone metastasis-related genes (BMRGs) were identified through a combination of differential gene expression analysis and WGCNA using GSE32269 and GSE77930 datasets. Consensus clustering analysis was employed to determine the significance of these genes in molecular subtyping of PCa. LASSO-Cox regression analysis was utilized to construct the bone metastasis-related prognostic gene signature (BMRPS). The predictive performance of BMRPS was assessed using ROC curves, Kaplan-Meier survival curves, and a predictive nomogram. The immune landscape heterogeneity of subgroups was analyzed using CIBERSORT, ESTIMATE, and xCell algorithms. Drug sensitivity and molecular docking analysis were performed to identify drugs associated with BMRPS. Results:Forty-four BMRGs associated with the prognosis of PCa were identified. Consensus clustering revealed the pivotal role of these genes in stratifying PCa into three distinct prognostic clusters. The BMRPS, consisting of 14 BMRGs, demonstrated excellent predictive accuracy for prognosis and served as an independent prognostic factor in PCa. BMRPS effectively predicted the overall survival of bone metastatic PCa and differentiated bone metastasis from other metastatic types. BMRPS showed a close correlation with the immune landscape and immunotherapeutic response biomarkers. Additionally, BMRPS was associated with anti-androgen resistance, and AZD8186 was identified as a potential BMRPS-related drug that holds promise for personalized treatment in PCa. Conclusion:BMRPS facilitates the prediction of prognosis and resistance to anti-androgens in PCa. It also offers insights into the molecular mechanisms of bone metastasis and aids in drug selection for the treatment of PCa.

Purpose Despite the proven clinical benefits of cytokine therapy in cancer treatment, systemic administration of cytokines such as IL-12 is constrained by dose-limiting toxicities and short half-lives. To address these challenges, we explored a localized cytokine delivery strategy using engineered neoantigen-reactive T (NRT) cells as carriers in a murine model of osteosarcoma.Materials and Methods We used a neoantigen from K7M2 osteosarcoma cells to retrovirally transduce NRT cells to express an inducible form of IL-12. We evaluated the engineered NRT cells' antitumor activity and the production of IL-12 and IFN-gamma upon in vitro co-culture with tumor cells. We systemically administered NRT-IL-12 cells in a mouse model of osteosarcoma to assess their impact on tumor growth and survival.Results In vitro assays demonstrated that the engineered NRT cells exhibited enhanced antitumor activity and produced elevated levels of IL-12 and IFN-gamma. In the mouse model of osteosarcoma, systemic administration of NRT-IL-12 cells resulted in a significant reduction in tumor growth and an increase in survival rates compared to the administration of control NRT cells. Further analysis revealed that NRT-IL-12 cells induced a profound increase in CD8+ T-cell infiltration and a decrease in Treg cells within the tumor microenvironment.Conclusion Our study presents a novel and efficacious strategy for osteosarcoma immunotherapy by harnessing NRT cells as targeted cytokine delivery vehicles.

Background Bone metastasis signi fi cantly impact the prognosis of non-small cell lung cancer (NSCLC) patients, reducing their quality of life and shortening their survival. Currently, there are no effective tools for the diagnosis and risk assessment of early bone metastasis in NSCLC patients. This study employed machine learning to analyze serum indicators that are closely associated with bone metastasis, aiming to construct a model for the timely detection and prognostic evaluation of bone metastasis in NSCLC patients. Methods The derivation cohort consisted of 664 individuals with stage IV NSCLC, diagnosed between 2015 and 2018. The variables considered in this study included age, sex, and 18 speci fi c serum indicators that have been linked to the occurrence of bone metastasis in NSCLC. Variable selection used multivariate logistic regression analysis and Lasso regression analysis. Six machine learning methods were utilized to develop a bone metastasis diagnostic model, assessed with Area Under the Curve (AUC), Decision Curve Analysis (DCA), sensitivity, speci fi city, and validation cohorts. External validation used 113 NSCLC patients from the Medical Alliance (2019 - 2020). Furthermore, a prospective validation study was conducted on a cohort of 316 patients (2019 - 2020) who were devoid of bone metastasis, and followed-up for at least two years to assess the predictive capabilities of this model. The model ' s prognostic value was evaluated using Kaplan - Meier survival curves. Findings Through variable selection, 11 serum indictors were identi fi ed as independent predictive factors for NSCLC bone metastasis. Six machine learning models were developed using age, sex, and these serum indicators. A random forest (RF) model demonstrated strong performance during the training and internal validation cohorts, achieving an AUC of 0.98 (95% CI 0.95 - 0.99) for internal validation. External validation further con fi rmed the RF model ' s effectiveness, yielding an AUC of 0.97 (95% CI 0.94 - 0.99). The calibration curves demonstrated a high level of concordance between the anticipated risk and the observed risk of the RF model. Prospective validation revealed that the RF model could predict the occurrence of bone metastasis approximately 10.27 +/- 3.58 months in advance, according to the results of the SPECT. An online computing platform (https://bonemetastasis.shinyapps.io/shiny_cls_ 1model/) for this RF model is publicly available and free-to-use by doctors and patients. Interpretation This study innovatively employs age, gender, and 11 serological markers closely related to the mechanism of bone metastasis to construct an RF model, providing a reliable tool for the early screening and prognostic assessment of bone metastasis in NSCLC patients. However, as an exploratory study, the fi ndings require further validation through large-scale, multicenter prospective studies.

BACKGROUND:Hearing loss and lifestyle factors have been associated with cognitive impairment. We aimed to explore the joint association of combined healthy lifestyle factors and hearing loss with cognitive impairment, which has been scarcely studied. METHODS:This baseline study used data from the CHOICE-Cohort study (Chinese Hearing Solution for Improvement of Cognition in Elders). Hearing loss was assessed by the better-ear pure-tone average. A composite healthy lifestyle score was built based on never smoking, never drinking, regular physical activity, and a balanced diet. Cognitive impairment was diagnosed by the Mini-Mental State Examination score of less than 24. RESULTS:We included 17 057 participants aged 60 years or older in China (mean age 69.8 [standard deviation 6.2] years, 55.7% female). Among the participants, 48.3% (n = 8 234) had mild hearing loss, and 25.8% (n = 4 395) had moderate or greater hearing loss. The proportion of participants with healthy lifestyle scores of 0-1, 2, 3, and 4 was 14.9% (n = 2 539), 29.3% (n = 5 000), 37.4% (n = 6 386), and 18.4% (n = 3 132), respectively. About 29.6% (n = 5 057) participants had cognitive impairment. When compared to those with normal hearing and healthy lifestyle (scores of 3-4), participants with hearing loss plus unhealthy lifestyle (scores of 0-2) exhibited approximately twofold increased risk of cognitive impairment (odds ratio [OR] = 1.92, 95% confidence interval [CI] 1.70-2.18). Conversely, the risk was greatly attenuated by adherence to healthy lifestyle in individuals with hearing loss (OR = 1.57, 95% CI 1.40-1.76). CONCLUSIONS:Our findings demonstrated adherence to a broad range of healthy lifestyle factors was associated with a significantly lower risk of cognitive impairment among participants with hearing loss.

目的:对比分析姑息一线应用贝伐珠单抗联合化疗治疗KRAS突变型晚期左、右半结肠癌的临床疗效.方法:回顾性分析我院于2016年09月至2020年09月收治的83例一线应用贝伐珠单抗联合化疗治疗KRAS突变型晚期左、右半结肠癌患者,比较KRAS突变型左、右半结肠癌患者的临床特征、客观缓解率(ORR)、疾病控制率(DCR)、无进展生存时间(PFS)、18个月生存率、总生存时间(OS).结果:KRAS突变型晚期右半结肠癌(RSCC)患者合并不完全肠梗阻及体重减轻>5 kg所占比例更高(P=0.369、P=0.009),KRAS突变型晚期左半结肠癌(LSCRC)患者更易合并中重度以上的贫血(P=0.154).KRAS突变型LSCRC患者发生腹腔种植转移及远处器官转移的患者比例高于RSCC患者(P>0.05).KRAS突变型LSCRC患者与RSCC患者ORR分别为29.3％和28.6％(P=0.944),DCR分别为87.8％和90.5％(P=0.696).18个月生存率LSCRC组为80.05％,RSCC组为76.2％,差异无统计学意义(P=0.635).KRAS突变型LSCRC患者中位PFS差于RSCC患者(8个月vs 9.8个月,P=0.004),KRAS突变型LSCRC患者中位OS优于KRAS突变型RSCC患者中位OS(22.8个月vs 21个月,P=0.001).结论:姑息一线应用贝伐珠单抗联合化疗治疗KRAS基因突变型mCRC患者,左、右半结肠癌的远期疗效存在差异.

Age-related hearing loss (ARHL) signifies the bilateral, symmetrical, sensorineural hearing loss that commonly occurs in elderly individuals. Several studies have suggested a higher risk of dementia among patients diagnosed with ARHL. Although the precise causal association between ARHL and cognitive decline remains unclear, ARHL has been recognized as one of the most significant factors that can be modified to reduce the risk of developing dementia potentially. Mild cognitive impairment (MCI) typically serves as the initial stage in the transition from normal cognitive function to dementia. Consequently, the objective of our randomized controlled trial (RCT) is to further investigate whether the use of hearing aids can enhance cognitive function in older adults diagnosed with ARHL and MCI. This study is a parallel-arm, randomized controlled trial conducted at multiple centers in Shanghai, China. We aim to enlist a total of 688 older adults (age ≥ 60) diagnosed with moderate-to-severe ARHL and MCI from our four research centers. Participants will be assigned randomly to either the hearing aid fitting group or the health education group using block randomization with varying block sizes. Audiometry, cognitive function assessments, and other relevant data will be collected at baseline, as well as at 6, 12, and 24 months post-intervention by audiologists and trained researchers. The primary outcome of our study is the rate of progression to dementia among the two groups of participants. Additionally, various evaluations will be conducted to measure hearing improvement and changes in cognitive function. Apart from the final study results, we also plan to conduct an interim analysis using data from 12-month follow-up. In recent years, there has been a notable lack of randomized controlled trials (RCTs) investigating the possible causal relationship between hearing fitting and the improvement of cognitive function. Our findings may demonstrate that hearing rehabilitation can be a valuable tool in managing ARHL and preventing cognitive decline, which will contribute to the development of a comprehensive framework for the prevention and control of cognitive decline. Chinese Clinical Trial Registry chictr.org.cn ChiCTR2000036139. Registered on 21 August 2020.

>Noise-induced hearing loss (NIHL) is one of the most common types of hearing loss and contributes to significant morbidity worldwide,as well as being the major cause of age-related hearing loss and tinnitus.The World Health Organization (WHO) estimates that 10%of the world's population is exposed to sound levels that could potentially cause NIHL[1].It has been widely accepted that NIHL is a multifactorial genetic disorder with estimated heritability of 36%to 40%[2].Ideally,identification of the modifying genetic variants would help protect individuals at high risk of NIHL.For decades,many geneticists have searched for genes that either predispose or protect from NIHL.Howeve r,the majority of genetic studies for NIHL were conducted in independent known candidate genes [3].

骨肉瘤是儿童及青少年最常见的骨恶性肿瘤,其特点为侵袭性强、易发生肺部转移,治疗较为困难.虽然传统外科手术联合术前新辅助及术后辅助化疗可提高患者 5 年生存率,局限期骨肉瘤患者 5 年生存率可达到 70％,但对于进展期、转移、复发的骨肉瘤患者而言,5 年生存率仅为 20％.骨肉瘤治疗方式在过去 30 年间未得到突破性进展,化疗耐药是其中的一大主要原因.为了解决化疗"瓶颈期"问题,学者们相继提出将靶向治疗作为一种新的生物治疗方法应用于骨肉瘤,并取得了一些成果.本文综述了目前晚期骨肉瘤靶向治疗的研究进展.

Osteosarcoma is one of the most common malignant bone tumors with high chemoresistance and poor prognosis, exhibiting abnormal gene regulation and epigenetic events. Methotrexate (MTX) is often used as a primary agent in neoadjuvant chemotherapy for osteosarcoma; However, the high dosage of methotrexate and strong drug resistance limit its therapeutic efficacy and application prospects. Studies have shown that abnormal expression and dysfunction of some coding or non-coding RNAs (e.g., DNA methylation and microRNA) affect key features of osteosarcoma progression, such as proliferation, migration, invasion, and drug resistance. Comprehensive multi-omics analysis is critical to understand its chemoresistant and pathogenic mechanisms. Currently, the network analysis-based non-negative matrix factorization (netNMF) method is widely used for multi-omics data fusion analysis. However, the effects of data noise and inflexible settings of regularization parameters affect its performance, while integrating and processing different types of genetic data is also a challenge. In this study, we introduced a novel adaptive total variation netNMF (ATV-netNMF) method to identify feature modules and characteristic genes by integrating methylation and gene expression data, which can adaptively choose an anisotropic smoothing scheme to denoise or preserve feature details based on the gradient information of the data by introducing an adaptive total variation constraint in netNMF. By comparing with other similar methods, the results showed that the proposed method could extract multi-omics fusion features more effectively. Furthermore, by combining the mRNA and miRNA data of methotrexate (MTX) resistance with the extracted feature genes, four genes, Carboxypeptidase E (CPE), LIM, SH3 protein 1 (LASP1), Pyruvate Dehydrogenase Kinase 1 (PDK1) and Serine beta-lactamase-like protein (LACTB) were finally identified. The results showed that the gene signature could reliably predict the prognostic status and immune status of osteosarcoma patients.

Nocardia rubra cell-wall skeleton (Nr-CWS) is reported as an external immunotherapeutic enhancer with the advantage of antitumor effect on human cancers. However, the immune regulatory role of Nr-CWS is not fully illustrated. We studied mouse CD4 + T lymphocytes isolated from mice spleen were induced by Nr-CWS and observed that the differentiation of Th1 CD4 + T cells and the cytokines of IL-2, TNF-α, IFN-γ were all enhanced by Nr-CWS. Furthermore, RNA sequencing was conducted to investigate the different mRNA profiling induced by Nr-CWS. We observed that paired box 8 (PAX8) was significantly up-regulated in Nr-CWS-treated Th1 cells compared to control. As a transcription factor, chromatin immunoprecipitation sequencing was carried out to study the genome-wide distribution of PAX8. Interestingly, we found that the binding domain of PAX8 was elevated by Nr-CWS, and the target genes associated with these binding sites showed a positive correlation between their transcription and PAX8 binding strength. Finally, we determined that Nr-CWS could enhance the activity of the PI3 K/Akt signaling pathway. Akt agonist could mimic the effect of Nr-CWS for PAX8 up-regulation, while Akt inhibitor compromised the expression of PAX8. Taken together, we determined a novel role of Nr-CWS in boosting the activity of Th1 maturation via the PI3 K/Akt/PAX8 axis. Keywords Nocardia rubra cell-wall skeleton , PAX8 , PI3 K/Akt , Th1 differentiation , CD4 , T cells

目的:探讨多西他赛联合红色诺卡氏菌细胞壁骨架(Nocardia rubra cell-wall skeleton,Nr-CWS)二线解救治疗晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)的疗效及其对患者免疫功能及生活质量评分的影响.方法:回顾性分析46例接受多西他赛联合Nr-CWS二线解救治疗的晚期NSCLC患者的临床资料.治疗方案为:单药多西他赛75 mg/m2静脉滴注,d 1,每隔21d重复1次;Nr-CWS400μg皮下注射,d 1,每隔7 d重复1次,直至化疗结束.所有患者均至少接受2个周期的治疗并根据实体瘤临床疗效评价标准评价疗效.检测治疗前后患者血液中的CD4+、CD8+和CD4+CD8+T淋巴细胞及自然杀伤(natural killer,NK)细胞水平,观察并比较治疗前和治疗2、4及6个周期后患者的生存质量评分和不良反应的发生情况.结果:46例入组患者中,完全缓解者有0例,部分缓解者有8例,疾病稳定者有24例;本研究治疗方案的客观有效率为17.4％,疾病控制率为69.6％;患者的中位无进展生存期为19周(95％置信区间:17.8～20.2周),中位总生存期为10个月(95％置信区间:9.5～10.9个月);治疗后患者血液中的CD4+、CD4+CD8+T淋巴细胞及NK细胞水平比治疗前均明显升高(P＜0.05);治疗2、4和6个周期后患者的生活质量评分明显高于治疗前(P＜0.05),患者的生活质量有明显改善;未观察到由Nr-CWS所致的严重不良反应.结论:Nr-CWS联合多西他赛二线解救治疗晚期非小细胞肺癌患者可以取得较好的疗效,能有效改善患者的免疫功能,提高生活质量,值得临床推广.

Nocardia rubra cell-wall skeleton (Nr-CWS) is reported as an external immunotherapeutic enhancer with the advantage of antitumor effect on human cancers. However, the immune regulatory role of Nr-CWS is not fully illustrated. We studied mouse CD4+ T lymphocytes isolated from mice spleen were induced by Nr-CWS and observed that the differentiation of Th1 CD4+ T cells and the cytokines of IL-2, TNF-α, IFN-γ were all enhanced by Nr-CWS. Furthermore, RNA sequencing was conducted to investigate the different mRNA profiling induced by Nr-CWS. We observed that paired box 8 (PAX8) was significantly up-regulated in Nr-CWS-treated Th1 cells compared to control. As a transcription factor, chromatin immunoprecipitation sequencing was carried out to study the genome-wide distribution of PAX8. Interestingly, we found that the binding domain of PAX8 was elevated by Nr-CWS, and the target genes associated with these binding sites showed a positive correlation between their transcription and PAX8 binding strength. Finally, we determined that Nr-CWS could enhance the activity of the PI3 K/Akt signaling pathway. Akt agonist could mimic the effect of Nr-CWS for PAX8 up-regulation, while Akt inhibitor compromised the expression of PAX8. Taken together, we determined a novel role of Nr-CWS in boosting the activity of Th1 maturation via the PI3 K/Akt/PAX8 axis.

Background: The definition of notched audiogram for noise-induced hearing loss (NIHL) is presently based on clinical experience, but audiometric phenotypes of NIHL are highly heterogeneous. The data-driven clustering of subtypes could provide refined characteristics of NIHL, and help identify individuals with typical NIHL at diagnosis.Methods: This cross-sectional study initially recruited 12,218 occupational noise-exposed employees aged 18–60 years from two factories of a shipyard in Eastern China. Of these, 10,307 subjects with no history of otological injurie or disease, family history of hearing loss, or history of ototoxic drug use were eventually enrolled. All these subjects completed health behavior questionnaires, cumulative noise exposure (CNE) measurement, and pure-tone audiometry. We did data-driven cluster analysis (k-means clustering) in subjects with hearing loss audiograms (n = 6,599) consist of two independent datasets (n = 4,461 and n = 2,138). Multinomial logistic regression was performed to analyze the relevant characteristics of subjects with different audiometric phenotypes compared to those subjects with normal hearing audiograms (n = 3,708).Results: A total of 10,307 subjects (9,165 males [88.9%], mean age 34.5 [8.8] years, mean CNE 91.2 [22.7] dB[A]) were included, 3,708 (36.0%) of them had completely normal hearing, the other 6,599 (64.0%) with hearing loss audiograms were clustered into four audiometric phenotypes, which were replicable in two distinct datasets. We named the four clusters as the 4–6 kHz sharp-notched, 4–6 kHz flat-notched, 3–8 kHz notched, and 1–8 kHz notched audiogram. Among them, except for the 4–6 kHz flat-notched audiogram which was not significantly related to NIHL, the other three phenotypes with different relevant characteristics were strongly associated with noise exposure. In particular, the 4–6 kHz sharp-notched audiogram might be a typical subtype of NIHL.Conclusions: By data-driven cluster analysis of the large-scale noise-exposed population, we identified three audiometric phenotypes associated with distinct NIHL subtypes. Data-driven sub-stratification of audiograms might eventually contribute to the precise diagnosis and treatment of NIHL.

It is widely accepted that even a single acute noise exposure at moderate intensity that induces temporary threshold shift (TTS) can result in permanent loss of ribbon synapses between inner hair cells and afferents. However, effects of repeated or chronic noise exposures on the cochlear synapses especially medial olivocochlear (MOC) efferent synapses remain elusive. Based on a weeklong repeated exposure model of bandwidth noise over 2-20 kHz for 2 hours at seven intensities (88 to 106 dB SPL with 3 dB increment per gradient) on C57BL/6J mice, we attempted to explore the dose-response mechanism of prolonged noise-induced audiological dysfunction and cochlear synaptic degeneration. In our results, mice repeatedly exposed to relatively low-intensity noise (88, 91, and 94 dB SPL) showed few changes on auditory brainstem response (ABR), ribbon synapses, or MOC efferent synapses. Notably, repeated moderate-intensity noise exposures (97 and 100 dB SPL) not only caused hearing threshold shifts and the inner hair cell ribbon synaptopathy but also impaired MOC efferent synapses, which might contribute to complex patterns of damages on cochlear function and morphology. However, repeated high-intensity (103 and 106 dB SPL) noise exposures induced PTSs mainly accompanied by damages on cochlear amplifier function of outer hair cells and the inner hair cell ribbon synaptopathy, rather than the MOC efferent synaptic degeneration. Moreover, we observed a frequency-dependent vulnerability of the repeated acoustic trauma-induced cochlear synaptic degeneration. This study provides a sight into the hypothesis that noise-induced cochlear synaptic degeneration involves both afferent (ribbon synapses) and efferent (MOC terminals) pathology. The pattern of dose-dependent pathological changes induced by repeated noise exposure at various intensities provides a possible explanation for the complicated cochlear synaptic degeneration in humans. The underlying mechanisms remain to be studied in the future.

Auditory brainstem response (ABR) serves as an objective indication of auditory perception at a given sound level and is nowadays widely used in hearing function assessment. Despite efforts for automation over decades, ABR threshold determination by machine algorithms remains unreliable and thereby one still relies on visual identification by trained personnel. Here, we described a procedure for automatic threshold determination that can be used in both animal and human ABR tests. The method terminates level averaging of ABR recordings upon detection of time-locked waveform through cross-correlation analysis. The threshold level was then indicated by a dramatic increase in the sweep numbers required to produce “qualified” level averaging. A good match was obtained between the algorithm outcome and the human readouts. Moreover, the method varies the level averaging based on the cross-correlation, thereby adapting to the signal-to-noise ratio of sweep recordings. These features empower a robust and fully automated ABR test.

Background: Melanoma accounts for more than 80% of deaths from all dermatologic cancers, mainly due to its widespread metastasis. High intensity focused ultrasound (HIFU) is a promising technique for cancer therapy. Here, we investigated the efficacy of HIFU against melanoma and the underlying mechanisms. Methods: A melanoma allograft mouse model was established to examine the tumor progression and survival rate. Anti-tumor immunity was determined by measuring cytokines, regulatory T cells (Tregs), Th17 cells and CD8+ effector T cells. Western blot, qPCR, RNAi and luciferase assay were performed to confirm the expression and regulation of microRNA (miR)-9-5p and transforming growth factor beta (TGF-β). Results: HIFU exposure significantly suppressed melanoma growth and metastasis by activating interferon gamma (IFN-γ) secretion, inhibiting Tregs and Th17 cells, and stimulating CD8+ effector T cells. TGF-β was a direct target of miR-9-5p. The anti-tumor effect of HIFU might be mediated through the miR-9-5p/TGF-β pathway. Conclusion: HIFU activates anti-tumor response and alters tumor microenvironment, which may serve as a potential therapeutic strategy for melanoma treatment.