Extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) have emerged as a new mode of intercellular crosstalk and are responsible for many of the therapeutic effects of MSCs. To promote the application of MSC-EVs, recent studies have focused on the manipulation of MSCs to improve the production of EVs and EV-mediated activities. The current paper details an optimization method using non-invasive low-intensity pulsed ultrasound (LIPUS) as the stimulation for improving oral MSC-EV production and effectiveness. Stem cells from apical papilla (SCAP), a type of oral mesenchymal stem cell, displayed intensity-dependent pro-osteogenic and anti-inflammatory responses to LIPUS without significant cytotoxicity or apoptosis. The stimuli increased the secretion of EVs by promoting the expression of neutral sphingomyelinases in SCAP. In addition, EVs from LIPUS-induced SCAP exhibited stronger efficacy in promoting the osteogenic differentiation and anti-inflammation of periodontal ligament cells in vitro and alleviating oral inflammatory bone loss in vivo. In addition, LIPUS stimulation affected the physical characteristics and miRNA cargo of SCAP-EVs. Further investigations indicated that miR-935 is an important mediator of the pro-osteogenic and anti-inflammatory capabilities of LIPUS-induced SCAP-EVs. Taken together, these findings demonstrate that LIPUS is a simple and effective physical method to optimize SCAP-EV production and efficacy.

Allergic rhinitis (AR) is a common inflammation that affects many people globally. Quercetin has anti-allergic biological activity in AR. Here, we aimed to explore the effects of quercetin on type 1 helper T (Th1)/Th2 and regulatory T cells (Treg)/Th17 balance. We established an ovalbumin (OVA)-induced mouse model and orally administered 20, 35, and 50 mg/kg/day quercetin. The nasal symptoms of mice were observed. The immunoglobulin levels, Treg/Th17-related factors, and pro-inflammatory factors were examined by ELISA. The differentiated inflammation cells were visualized using the diff-quick staining assay. The nasal histopathology was evaluated using H&E, periodic acid Schiff (PAS), and Giemsa staining assay. The results showed that quercetin attenuated OVA-induced rubbing and sneezing. Quercetin reduced IgE, IgG1, histamine, and increased IgG2 in serum. The number of differentiated inflammation cells and goblet cells in tissues that elevated by OVA was reduced by quercetin. Moreover, OVA increased the Treg cell percentage, the levels of IL-17, TGF-beta, IL-6, TNF-alpha, and decreased Th17 cell percentage, IL-10 and FOXP3 levels, while quercetin abrogated their levels induced by OVA. Additionally, quercetin inactivated the NF-kappa B pathway. Taken together, quercetin attenuated AR symptoms by balancing the Th1/Th2, Treg/Th17 ratios, and inactivating the NF-kappa B pathway. The results suggested that quercetin may use for AR treatment.

Aim: T-regulatory (Treg)/T-helper (Th) 17 imbalance contributes to the pathogenesis of allergic rhinitis (AR). Long non-coding RNAs (lncRNAs) participate in the progression of AR. Herein, the effect of lncRNA JP X on Treg/Th17 balance in AR was explored.Methods: CD4+ T cells were isolated from patients with AR and healthy control. The percentage of Treg and Th17 cells were examined by flow cytometry. The levels of JP X, miR-378g, CCL5, T GF-β, and IL-17A were tested using qRT-P CR. The protein expression of Foxp3 and RORγt was measured by western blot.Results: The data showed that an imbalance of Treg/Th17 was associated with AR. Upregulation of JP X was found in AR, and knockdown of which improved the imbalance of Treg/Th17. Furthermore, JP X functioned as a sponge of miR-378g to upregulate CCL5. Inhibition of miR-378g reversed the effects on Treg/Th17 induced by silencing of JP X. Moreover, overexpression of CCL5 reversed miR-378g-induced effects.Conclusion: In conclusion, depletion of JP X promoted Treg/Th17 balance in AR via regulating the miR-378g/CCL5 axis. The findings provided a novel therapeutic insight for AR.

Purpose:Methacrylic anhydride-modified gelatin (GelMA) hydrogels exhibit many beneficial biological features and are widely studied for bone tissue regeneration. However, deficiencies in the mechanical strength, osteogenic factors and mineral ions limit their application in bone defect regeneration. Incorporation of inorganic fillers into GelMA to improve its mechanical properties and bone regenerative ability has been one of the research hotspots.Methods:In this work, hydroxyapatite nanofibers (HANFs) were prepared and mineralized in a simulated body fluid to make their components and structure more similar to those of natural bone apatite, and then different amounts of mineralized HANFs (m-HANFs) were incorporated into the GelMA hydrogel to form m-HANFs/GelMA composite hydrogels. The physicochemical properties, biocompatibility and bone regenerative ability of m-HANFs/GelMA were determined in vitro and in vivo.Results:The results indicated that m-HANFs with high aspect ratio presented rough and porous surfaces coated with bone-like apatite crystals. The incorporation of biomimetic m-HANFs improved the biocompatibility, mechanical, swelling, degradation and bone regenerative performances of GelMA. However, the improvement in the performance of the composite hydrogel did not continuously increase as the amount of added m-HANFs increased, and the 15m-HANFs/GelMA group exhibited the best swelling and degradation performances and the best bone repair effect in vivo among all the groups.Conclusion:The biomimetic m-HANFs/GelMA composite hydrogel can provide a novel option for bone tissue engineering in the future; however, it needs further investigations to optimize the proportions of m-HANFs and GelMA for improving the bone repair effect.

Background and Objectives Periodontitis, the most common chronic inflammation characterized by persistent alveolar bone resorption in the periodontitis, affects almost half of the adult population worldwide. Oxidative stress is one of the pathophysiological mechanisms underlying periodontitis, which affects the occurrence and development of periodontitis. Exosomes are increasingly recognized as vehicles of intercellular communication and are closely related to periodontitis. However, the effects of oxidative stress on exosome secretion and the specific mechanisms remain elusive in human periodontal ligament cells (hPDLCs). The relationship between exosome secretion and the osteogenic differentiation of hPDLCs also needs to be investigated. Methods Isolated PDLSCs were identified using flow cytometry. Osteogenesis was measured using alizarin red staining and ALP staining. Expression of exosomal markers and PRMT1 was analyzed using western blot. Immunofluorescence was used to measure exosome uptake and the expression of EEA1. Results The secretion capacity of exosomes was markedly suppressed under oxidative stress. Protein arginine methyltransferase 1 (PRMT1) has been strongly associated with both oxidative stress and inflammation, and PRMT1 was significantly upregulated under oxidative stress conditions. Lentivirus-mediated overexpression of PRMT1 caused a significant reduction in the secretion of exosomes, but multivesicular bodies (MVBs) containing a large number of intraluminal vesicles (ILVs) were increased. Rab11a and Rab27a expression, which mediate MVBs fusion with cell membranes, decreased, although this phenomenon was restored after knocking down PRMT1 expression under oxidative stress. Conclusions These results indicated that PRMT1 mediated a decrease in exosome secretion of hPDLCs. The decrease in Rab11a and Rab27a leads to a large accumulation of MVBs in cells and is one of the main reasons for impaired exosome secretion. The decrease in osteogenic differentiation of hPDLCs caused by H2O2 may originate in part from the inhibition of exosome secretion.

Abstract Objectives To explore the long-term efficacy and possible mechanism of endolymphatic sac decompression (ESD) in the treatment of Meniere’s disease(MD) by electrocochleography. Design Prospective cohort study. Setting “Blinded for review” Participants A total of 85 patients with MD who underwent unilateral ESD between June 2015 and November 2019 at“Blinded for review”were enrolled. Main outcome measures The Dizziness Handicap Inventory (DHI), Tinnitus Handicap Inventory (THI), pure tone audiometry, and electrocochleography were used for assessment. The mean follow-up time was 26 months (range: 7–60 months). Results The number of vertigo episodes was significantly reduced following ESD compared to before the surgery (P<0.005), and all patients achieved complete or basic control of vertigo as evidenced by a decrease in DHI score (P<0.005). THI scores of patients with tinnitus were also lower after as compared to before ESD (P<0.005), whereas no significant change in average hearing threshold of the affected side was observed (P>0.05). The cochlear summating potential (SP)/auditory nerve action potential (AP) area ratio in the electrocochleogram of the affected side was negatively correlated with DHI score (rs=−0.159, P=0.0074). Conclusions ESD achieved effective long-term control of vertigo in MD patients and improved the associated tinnitus without any obvious damage to hearing. Electrocochleography was useful for postoperative monitoring; the SP/AP area ratio of the affected side was closely related to the improvement of postoperative vertigo, possibly reflecting greater relief of pressure in the endolymphatic sac. Keypoints •ESD provided long-term control of vertigo in MD patients and improved the accompanying tinnitus without obviously damaging hearing. •ECochG is useful for monitoring the postoperative outcome of ESD. •The SP/AP area ratio of the affected side was closely related to the improvement of postoperative vertigo. •The endolymphatic sac is a “decompression reservoir” of the membranous labyrinth. •ESD may provide space for expansion of the endolymphatic sac and relieve pressure within the membranous labyrinth caused by endolymphatic hydrops, thereby reducing the risk of sac rupture and restoring pressure balance to reduce the sensation of vertigo.

AIM:To explore the role of C-reactive protein (CRP) in periodontitis and diabetes and its mechanism in alveolar bone homeostasis.MATERIALS AND METHODS:In vivo, normal, and Crp knockout (KO) rats were randomly divided into control, diabetes, periodontitis, and diabetes and periodontitis groups, respectively. The diabetes model was established using a high-fat diet combined with streptozotocin injection. The periodontitis model was established by ligature combined with lipopolysaccharide (LPS) injection. Alveolar bones were analysed using micro-computed tomography, histology, and immunohistochemistry. In vitro, human periodontal ligament cells (hPDLCs) were treated with LPS and high glucose. CRP knockdown lentivirus or CRP overexpression adenovirus combined with a PI3K/AKT signalling inhibitor or agonist were used to explore the regulatory mechanism of CRP in osteogenesis and osteoclastogenesis of hPDLCs, as evidenced by alkaline phosphatase staining, Western blot, and quantitative polymerase chain reaction.RESULTS:In periodontitis and diabetes, CRP KO decreased the alveolar bone loss and the expression levels of osteoclastogenic markers, while increasing the expression levels of osteogenic markers. CRP constrained osteogenesis while promoting the osteoclastogenesis of hPDLCs via PI3K/AKT signalling under high glucose and pro-inflammatory conditions.CONCLUSIONS:CRP inhibits osteogenesis and promotes osteoclastogenesis via PI3K/AKT signalling under diabetic and pro-inflammatory conditions, thus perturbing alveolar bone homeostasis.

目的 本研究旨在从Th1/Th2及Th17/Treg平衡轴的角度探讨PM2.5暴露及细菌溶解产物Broncho-Vaxom(BV)治疗与健康大鼠气道炎症的关系.方法 PM2.5暴露组鼻内滴注PM2.5混悬液,口服生理盐水;BV治疗组暴露与PM2.5暴露组相同,口服BV溶液;对照组均用生理盐水代替.对大鼠鼻黏膜进行组织病理学检查;用流式细胞仪和酶联免疫吸附试验(ELISA)检测Th1/Th2/Th17/Treg细胞的表达和功能.结果 PM2.5暴露组大鼠鼻黏膜出现炎症细胞浸润;Th2细胞和Th17细胞显著上调,Treg细胞显著下调;血清中相关细胞因子IL-4、IL-5、IL-13及IL-17显著升高,IFN-γ和IL-10显著下降.BV干预后鼻黏膜炎性细胞浸润减少、Th细胞相关免疫失衡减轻.结论 PM2.5可诱导健康大鼠产生气道炎症,PM2.5暴露所诱导的气道炎症可能与Th1/Th2和Th17/Treg细胞免疫失衡有关,BV可减轻PM2.5诱导的气道炎症及Th细胞相关免疫失衡.

BACKGROUND We aimed to explore the correlation between patients' sigmoid sinusoidal tinnitus (SST) and low-frequency sensorineural hearing loss (LFSHL) and illustrate the underlying mechanism. MATERIAL AND METHODS Seven healthy volunteers with normal hearing were subjected to 125-, 250-, and 500-Hz pure sound and different white noise-masking intensities. A retrospective analysis was made on the clinical data and postoperative follow-up data of 59 patients with SST in the First Affiliated Hospital of Chongqing Medical University. The patients' sex, age, chief complaints, affected site, concomitant symptoms, course of disease, pure-tone audiometry (PTA) results, tinnitus discomfort loudness scale results, imaging examination, and complications were collected. RESULTS The results of the simulation experiment showed that the threshold of each frequency segment was higher after noise masking than before masking; the intensity of noise masking was positively correlated with hearing loss, and the changes of the hearing threshold of the 3 frequencies before and after masking were statistically significant (P<0.05). Fifty-nine patients with SST were documented between January 2015 and January 2020. After the operation, their low-frequency hearing was recovered to normal; 11 cases had significantly alleviated tinnitus and 9 cases were cured. CONCLUSIONS SST often causes corresponding pseudo-low-frequency hearing loss due to the noise-masking effect. The center frequency of tinnitus appears not to be 250-Hz or 500-Hz octave frequency of PTA, barring the detection of the pseudo-hearing loss in the audiometry chart of most patients. Surgery positively affects patients with SST, and the pseudo-LFSHL can be completely recovered after the operation as a result of tinnitus elimination.

1 病例资料 患者,女,9岁,汉族.自幼右耳耳廓畸形并外耳道闭锁.5岁时因腹痛就诊于当地医院,腹部超声检查发现右侧肾缺如,患儿发育较同龄人一致,智力正常,言语发育好.患儿为第一胎,同卵双胎之一,孕38周足月生产,出生时体重2.55kg,出生无缺氧史,无明显黄疸等.其孪生姐姐无耳廓畸形及肾缺如.母亲孕5月有感染史,无特殊用药,无糖尿病等代谢疾病史.父亲左足第四、第五趾并趾畸形,简单查体未发现听力障碍及颜面部畸形,未行相关听力学检查.否认耳廓畸形、耳聋及肾缺如家族史.

Objective: To analyze the clinical characteristics and identify the causative gene of a case with congenital deafness. Methods: Detailed medical history and clinical examination of a 4-year-old male child with congenital deafness were conducted in the First Affiliated Hospital of Army Military Medical University in June 2016. He was diagnosed with sensorineural deafness. The venous blood of the child and his parents was drawn, and genomic DNA was extracted. Proband's DNA was performed with targeted capture of high-throughput sequencing, then Sanger sequencing was used to verify the suspected mutation and segregation in this pedigree. According to the genetic diagnosis of the proband's deafness, ophthalmic examinations were performed. Genetic prenatal diagnosis was performed when the proband's mother was pregnant again. Results: The patient was detected with p.Trp1466Ter/p.Tyr2042Ter compound heterozygous mutations of MYO7A gene with targeted high-throughput sequencing. The mutation of p.Trp1466Ter was a reported mutation, while p.Tyr2042Ter has not been reported. In addition to congenital deafness, retinitis pigmentosa was also found by ophthalmologic examination, and the patient was clinically diagnosed with Usher syndrome type 1. Amniocentesis and fetal DNA sequencing were performed on the repregnancy fetus of this family at 18 weeks of gestation. The heterozygous mutation of MYO7A gene p.Tyr2042Ter was found, and the other allele was the wild type, indicating that the child will not exhibit clinical manifestations of Usher syndrome type 1. Indeed, the second child passed neonatal hearing screening. Conclusions: The clinical features and genetic variants were delineated in this family with Usher syndrome type 1. The results of the current study have enriched the phenotype and genotype data of the disease and provided a basis for genetic counseling.

目的:基于AhR信号通路激活导致Th免疫失衡,探讨PM2.5暴露及细菌溶解产物(BV)治疗对小鼠气道炎症的作用及机制.方法:30只BALB/c雌性小鼠随机分为PM2.5暴露组、BV干预组和对照组,每组10只.PM2.5暴露组和BV干预组小鼠气管滴注PM2.5混悬液,连续15 d,第16～90天,PM2.5组口服给予生理盐水,BV干预组口服给予BV溶液,对照组给予等量生理盐水.组织病理学检查评价气道炎症程度;ELISA检测血清相关炎症因子水平;Western blot和RT-qPCR检测AhR表达.结果:PM2.5暴露后,小鼠气管黏膜出现大量炎症细胞浸润,血清IL-4、IL-5、IL-13、IL-17、IL-33水平显著升高,肺组织中AhR蛋白和mRNA表达增加.BV治疗后,小鼠血清炎症因子失衡减轻,气道黏膜炎症细胞浸润缓解.结论:PM2.5暴露可导致健康小鼠气道炎症,可能与AhR通路激活有关,BV可通过调节免疫反应减轻PM2.5诱导的气道炎症.

Background: There is no effective treatment for idiopathic tinnitus. Both acoustic therapy and acupuncture have been used in the treatment of idiopathic tinnitus, but the clinical efficacy is quite different. For there is no clinical study combining the 2, the purpose of this randomized controlled trial is to evaluate the effectiveness and safety of acoustic therapy combined with acupuncture in the treatment of idiopathic tinnitus. Methods: This is a prospective randomized controlled trial to study the effectiveness and safety of acoustic therapy combined with acupuncture in the treatment of idiopathic tinnitus, and is approved by the clinical research ethics committee of our hospital. The patients are randomly divided into one of 2 treatment options: (A) acoustic therapy combined with acupuncture group and (B) simple acupuncture group. Patients, doctors, nurses, and data collection assistants are blinded to group allocation. Observation indicators include: 1. Tinnitus Disability Scale; 2. Loudness visual analog scale; 3. Adverse reactions. Data is analyzed using the statistical software package SPSS version 25.0 (Chicago, IL). Discussion: This protocol will evaluate the efficacy and safety of acoustic therapy combined with acupuncture in the treatment of idiopathic tinnitus. The results of this experiment will provide clinical evidence for the use of acoustic therapy combined with acupuncture in the treatment of idiopathic tinnitus. Ethics and dissemination: Private information from individuals will not be published. This systematic review also does not involve endangering participant rights. Ethical approval was not required. OSF Registration number: DOI 10.17605/OSF.IO/87VFB.

Abstract Background Hearing loss is a common disease globally, and more than 50% of the cases are genetic. Autosomal recessive nonsyndromic hearing loss (ARNSHL) is one of the most common types of hereditary hearing loss. Here, a novel MYO15A missense mutation was identified in a Chinese family with ARNSHL, using targeted genetic sequencing and Sanger sequencing. Case presentation: A 6-year-old girl with congenital nonsyndromic sensorineural deafness was presented from the First Affiliated hospital of Chongqing Medical University, China. We used targeted region sequencing, Sanger sequencing, functional prediction, and three-dimensional protein structure modeling to identify and verify the genes responsible for deafness in the family. Conclusions We found pathogenic compound heterozygous mutations in MYO15A, including a novel missense mutation, c.6353T > C (p.Leu2118Pro). It could provide help not only for genetic counseling but also for further understanding of the functional role of MYO15A mutations.

Objectives: Sudden sensorineural hearing loss (SSNHL) is an emergency disease with undefined pathogenesis in the otolaryngology department. In our previous study, we found patients with SSNHL had lower serum concentration of Matrix metalloprotease 9 (MMP-9) than healthy controls, and the result was accordant with auto-immune diseases. This study aimed to reveal the correlation between changes in serum MMP-9 concentration following treatment with the outcomes of patients and to provide further evidence that immune disorder was the main pathogenesis of SSNHL. Design, setting, and participants: Fifty-two patients with SSNHL, hospitalized in The First Affiliated Hospital of Chongqing Medical University from March 2019 to August 2019, were enrolled. The serum concentration of MMP-9 was detected by enzyme-linked immunosorbent assay (ELISA). Main outcome measure: The mean concentration of MMP-9 before treatment was compared with the post-treatment concentration by the Mann-Whitney U test. The correlations between favorable outcomes of patients and clinical characteristics were measured with the Chi-squared test and binary multiple logistic regression analysis. Results: In treatment responders, mean serum concentration was elevated from 106.85±41.40ng/ml to 144.03±37.65 ng/ml following treatment (P＜0.001), while in non-responders it decreased from 132.09±59.21 ng/ml to 106.82±49.93 ng/ml (P=0.142). Changes in MMP-9 concentration was the only factor associated with favorable outcomes (P=0.008, OR=5.13, 95% CI: 1.53-17.28). Conclusions: Elevated MMP-9 concentration is a potential prognosis biomarker in patients with SSNHL. These findings are in line with auto-immune diseases and indicate immune disorder is mainly pathogenesis in SSNHL. Keywords Sudden sensorineural hearing loss; Matrix metalloprotease 9; Immune disorder; prognosis; Enzyme-linked immunosorbent assay

目的 分析老年性突发性耳聋的流行病学特征及其预后关系,为提高疗效提供临床思路.方法 回顾性分析2015年1月～2018年12月重庆医科大学附属第一医院突发性聋患者中≥65岁的临床病例229例(241耳),描述其流行病学特征,并探讨其与疗效的相关性.结果 老年突发性聋占所有突发性聋患者的10.7％.229例老年突发性聋患者中,初诊时间≤14 d的172例、＞14 d的57例;听力下降前后出现耳鸣者189例,以持续性嗡嗡声为主;有耳闷者85例;有眩晕或者头晕者80例;无明显诱因者占91.3％、有感冒病史者占4.4％、有劳累、压力大、情绪波动因素者占3％、其他因素占1.3％;伴高血压病者121例;伴糖尿病者65例;伴冠心病者42例;伴高血压、糖尿病、冠心病任一基础疾病者146例,占63.8％.241例患耳中,按耳聋程度分级:轻度27耳、中度44耳、重度67耳、极重度103耳;按听力曲线类型分类:低频下降型6耳、高频下降型24耳、平坦型111耳和全聋型100耳.老年突发性聋患者的性别、是否伴耳鸣、耳闷、头晕或眩晕对临床总有效率的影响不具有统计学意义(P＞0.05);不同初诊时间、不同听力曲线类型与患者的临床总有效率比较,差异有统计学意义(P＜0.05),即初诊时间越长,疗效越差.高频听力损失患者的疗效较全聋型差.结论 老年突发性聋患者虽多伴有基础疾病,但是否伴基础疾病与疗效无明显相关性,在就诊及时的情况下,临床总有效率较高,且高频听力损失患者的预后较全聋型差.

分析2例合并Treacher Collins综合征的人工耳蜗植入患儿的临床资料,探讨其围手术期策略与手术操作技巧,为提高手术成功率,防止并发症提供经验借鉴.对合并Treacher Collins综合征的人工耳蜗植入患者,术前详询病史与查体,全面评估气道,完善听力学、影像学检查,利于选择合适的手术侧别,制定最佳手术策略与补救措施.术中使用面神经监护仪可以避免面瘫等严重并发症发生,并有助于高效安全地完成手术.

音调性耳鸣是一种耳鸣频率可以被匹配出来的主观性耳鸣.目前大部分主观性耳鸣尚无有效的治疗方法,但针对音调性耳鸣的一种新的治疗方法——个性化切迹音乐治疗被提出,其疗效值得期待.本文就个性化切迹音乐治疗音调性耳鸣的原理、方法、临床疗效等进行综述,探讨其存在的问题及临床应用前景.

Objective To report clinical features and causes of misdiagnosed Vestibular Paroxysmia (VP). Meth-ods Ten patients with eventual diagnosis of VP or probable VP at the First Affiliated Hospital of Chongqing Medical University were included. Among them, only 1 was initially diagnosed with VP and the rest were misdiagnosed. Six of the 10 patients were diagnosed with VP and 4 with probable VP at discharge. Their diagnoses and treatment, medical his-tory and physical examination findings were reviewed. Results VP diagnosis is difficult and depends mainly on clinical manifestations, audiology and vestibular test results, as well as characteristic imaging features. Clinical features of VP include episodic vertigo attack with a duration of less than 1 minute, hypofunction on vestibular tests, hearing loss,ab-normal ABR results, and signs of neurovascular compression (NVC) in the internal auditory canal on MRI. Antiepileptic drugs are effective for VP. Conclusion VP can be easily misdiagnosed due to lack of specific clinical manifestations, Care should be taken to distinguish VP from other types of vertigo diseases, such as Meniere's disease, BPPV, vestibular migraine and sudden deafness with vertigo. Clinicians should increase their understanding of VP in order to improve its diagnosis and treatment.