Planar Cell Polarity in the Multiciliated Epithelial Lining of the Mouse Eustachian Tube.

OBJECTIVES: Planar cell polarity (PCP) signaling, essential for uniform alignment and directional beating of motile cilia, has been investigated in multiciliated epithelia. As a complex structure connecting the middle ear to the nasopharynx, the eustachian tube (ET) is important in the onset of ear-nose-throat diseases. However, PCP signaling, including the orientation that is important for ciliary motility and clearance function in the ET, has not been studied. We evaluated PCP in the ET epithelium. STUDY DESIGN: Morphometric examination of the mouse ET. METHODS: We performed electron microscopy to assess ciliary polarity in the mouse ET, along with immunohistochemical analysis of PCP protein localization in the ET epithelium. RESULTS: We discovered PCP in the ET epithelium. Motile cilia were aligned in the same direction in individual and neighboring cells; this alignment manifested as ciliary polarity in multiciliated cells. Additionally, PCP proteins were asymmetrically localized between adjacent cells in the plane of the ET. CONCLUSIONS: The multiciliated ET epithelium exhibits polarization, suggesting novel structural features that may be critical for ET function. LEVEL OF EVIDENCE: NA Laryngoscope, 134:3795-3801, 2024.

[The value of HRCT in predicting cerebrospinal fluid gusher during cochlear implantation].

Objective:To investigate the predictive value of temporal bone high-resolution CT（HRCT） multiplanar reconstruction（MPR） for cerebrospinal fluid（CSF） gusher during cochlear implantation in patients with inner ear malformation. Methods:The clinical data of 33 patients（36 ears） with inner ear malformation who underwent cochlear implantation were retrospectively analyzed. The predictive value of HRCT for cerebrospinal fluid gusher during cochlear implantation was evaluated. Results:The width of the cochlear foramen（P=0.024, OR=1.735） and the diameter of the inner auditory meatus（P=0.022, OR=6.119） were independent risk factors for CSF gusher during cochlear implantation. The area under the curve（AUC） of cochlear foramen width in predicting intraoperative gusher was 0.851, the sensitivity was 93.33%, and the specificity was 61.90%. The AUC of the upper and lower diameter of the internal auditory canal for predicting intraoperative gusher was 0.848, the sensitivity was 80.00%, and the specificity was 80.95%. The AUC of cochlear foramen width combined with the upper and lower diameters of the internal auditory meatus for predicting intraoperative gusher was 0.930, the sensitivity was 80.00%, and the specificity was 95.24%. Conclusion:Based on temporal bone HRCT, the prediction model of cochlear foramen width combined with the upper and lower diameter of the internal auditory canal has crucial predictive value for the "gusher" during cochlear implantation in patients with inner ear malformation.

The Chinese Zodiac-in-Noise Test: An Internet-Based Speech-in-Noise Test for Large-Scale Hearing Screening.

OBJECTIVES: Motivated by the growing need for hearing screening in China, the present study has two objectives. First, to develop and validate a new test, called the Chinese Zodiac-in-noise (ZIN) test, for large-scale hearing screening in China. Second, to conduct a large-scale remote hearing screening in China, using the ZIN test developed. DESIGN: The ZIN test was developed following a similar procedure as the digits-in-noise test but emphasizes the importance of consonant recognition by employing the 12 zodiac animals in traditional Chinese culture as speech materials. It measures the speech reception threshold (SRT) using triplets of Chinese zodiac animals in speech-shaped noise with an adaptive procedure. RESULTS: Normative data of the test were obtained in a group of 140 normal-hearing listeners, and the performance of the test was validated by comparisons with pure-tone audiometry in 116 listeners with various hearing abilities. The ZIN test has a reference SRT of -11.0 ± 1.6 dB in normal-hearing listeners with a test-retest variability of 1.7 dB and can be completed in 3 minutes. The ZIN SRT is highly correlated with the better-ear pure-tone threshold ( r = 0.82). With a cutoff value of -7.7 dB, the ZIN test has a sensitivity of 0.85 and a specificity of 0.94 for detecting a hearing loss of 25 dB HL or more at the better ear.A large-scale remote hearing screening involving 30,552 participants was performed using the ZIN test. The large-scale study found a hearing loss proportion of 21.0% across the study sample, with a high proportion of 57.1% in the elderly study sample aged over 60 years. Age and gender were also observed to have associations with hearing loss, with older individuals and males being more likely to have hearing loss. CONCLUSIONS: The Chinese ZIN test is a valid and efficient solution for large-scale hearing screening in China. Its remote applications may improve access to hearing screening and enhance public awareness of hearing health.

The role of Espin in the stereocilia regeneration and protection in Atoh1-overexpressed cochlear epithelium.

Hair cells (HCs) in mammals cannot spontaneously regenerate after damage. Atoh1 overexpression can promote HC regeneration in the postnatal cochlea, but the regenerated HCs do not possess the structural and functional characteristics of HCs in situ. The stereocilia on the apical surface of HCs are the first-level structure for sound conduction, and regeneration of functional stereocilia is the key basis for the reproduction of functional HCs. Espin, as an actin bundling protein, plays an important role in the development and structural maintenance of the stereocilia. Here, we found that the upregulation of Espin by AAV-ie was able to induced the aggregation of actin fibres in Atoh1-induced HCs in both cochlear organoids and explants. In addition, we found that persistent Atoh1 overexpression resulted in impaired stereocilia in both endogenous and newly formed HCs. In contrast, the forced expression of Espin in endogenous and regenerative HCs was able to eliminate the stereocilia damage caused by persistent Atoh1 overexpression. Our study shows that the enhanced expression of Espin can optimize the developmental process of stereocilia in Atoh1-induced HCs and can attenuate the damage to native HCs induced by Atoh1 overexpression. These results suggest an effective method to induce the maturation of stereocilia in regenerative HCs and pave the way for functional HC regeneration via supporting cell transdifferentiation.

Rps14 upregulation promotes inner ear progenitor proliferation and hair cell regeneration in the neonatal mouse cochlea.

Sensorineural hearing loss a result from hair cell damage, which is irreversible in mammals owing to the lack of hair cell regeneration, but recent researches have shown that Lgr5+ supporting cells are progenitors capable of regenerating hair cells. RPS14 (ribosomal protein S14) is a 40S ribosomal subunit component and is associated with erythrocyte differentiation, and in this study, we used a novel adeno-associated virus-inner ear system to upregulate Rps14 expression in cultured hair cell progenitors and observed an enhancement on their ability to proliferate and differentiate into hair cells. Similarly, Rps14 overexpression in the mice cochlea could promote supporting cells proliferation by activating the Wnt signalling pathway. In addition, over-expressing Rps14 induced hair cells regeneration in the organ of Corti, and lineage tracing showed that the new hair cells had transformed from Lgr5+ progenitors. In conclusion, our analysis reveals the potential role of Rps14 in driving hair cell regeneration in mammalian.

The natural history of OTOF-related auditory neuropathy spectrum disorders: a multicenter study.

Pathogenic variations in the OTOF gene are a common cause of hearing loss. To refine the natural history and genotype-phenotype correlations of OTOF-related auditory neuropathy spectrum disorders (ANSD), audiograms and distortion product otoacoustic emissions (DPOAEs) were collected from a diverse cohort of individuals diagnosed with OTOF-related ANSD by comprehensive genetic testing and also reported in the literature. Comparative analysis was undertaken to define genotype-phenotype relationships using a Monte Carlo algorithm. 67 audiograms and 25 DPOAEs from 49 unique individuals positive for OTOF-related ANSD were collected. 51 unique OTOF pathogenic variants were identified of which 21 were missense and 30 were loss of function (LoF; nonsense, splice-site, copy number variants, and indels). There was a statistically significant difference in low, middle, and high frequency hearing thresholds between missense/missense and LoF/missense genotypes as compared to LoF/LoF genotypes (average hearing threshold for low, middle and high frequencies 70.9, 76.0, and 73.4 dB vs 88.5, 95.6, and 94.7 dB) via Tukey's test with age as a co-variate (P = 0.0180, 0.0327, and 0.0347, respectively). Hearing declined during adolescence with missense/missense and LoF/missense genotypes, with an annual mid-frequency threshold deterioration of 0.87 dB/year and 1.87 dB/year, respectively. 8.5% of frequencies measured via DPOAE were lost per year in individuals with serial tests. Audioprofiling of OTOF-related ANSD suggests significantly worse hearing with LoF/LoF genotypes. The unique pattern of variably progressive OTOF-related autosomal recessive ANSD may be amenable to gene therapy in selected clinical scenarios.

Long-Term Outcomes of Cochlear Implantation in Irradiated Ears of Nasopharyngeal Carcinoma Patients.

OBJECTIVES: To evaluate the safety and feasibility of cochlear implantation (CI) in irradiated ears of nasopharyngeal carcinoma (NPC) patients. STUDY DESIGN: A retrospective study. METHODS: From 2008 to 2017, 10 adults with binaural severe or profound sensorineural hearing loss subsequent to radiotherapy for NPC underwent CI in our center. The mean follow-up was 63.2 months. Hearing and speech performance were evaluated pre- and postoperatively with audiometric and speech discrimination testing, Category of Auditory Performance, Speech Intelligibility Rating, and Chinese version of Nijmegen Cochlear Implant Questionnaire. RESULTS: According to the severity of radioactive osteomyelitis of temporal bone, patients were classified into three categories: mild, moderate, and severe. Patients in mild category underwent routine CI. Patients in moderate category underwent extended radical mastoidectomy and CI simultaneously. Patients in severe category underwent subtotal temporal bone resection, external auditory canal elimination, and CI simultaneously or by stage. There was no massive hemorrhage, facial paralysis, or nonunion of incision after operation. One suffered from radiation encephalopathy 13 months postoperatively; since then, the cochlear implant has been idle. The other nine patients demonstrated encouraging results of hearing and speech performance. CONCLUSION: CI for postirradiated ears of NPC is safe and feasible. Strict control of operative indications and extra care during surgery are required. Individual surgical plan should be made according to the severity of radioactive osteomyelitis. Choose one-stage surgery as much as possible in severe cases to avoid the risk of difficulty in locating round window in second-stage surgery. LEVEL OF EVIDENCE: 4 Laryngoscope, 131:649-655, 2021.

[The long term effect of malleostapedotomy in ossicular chain reconstruction].

Objective:To analysis the long-term outcomes of ossicular chain reconstruction using the malleostapedotomy（MT）. Method:A total of 11 patients（12 ears） underwent MT and their hearing levels were measured prior to surgery, 1 week and more than 1 year after surgery. The indications of MT were discussed and its safety and efficacy were evaluated in terms of the intra-operative findings, post-operative hearing and complications. Result:Among 11 patients（12 ears）, there were 1 patient（1 ear） with tympanosclerosis, 3 patients（4 ears） with ossicular chain deformity, 5 patients（5 ears） with otosclerosis and 2 patients（2 ears） with localized cholesteatoma of the middle ear. No cases of bone conduction hearing loss（more than 10 dB） were observed within 2 weeks after surgery while four patients suffered from short-term vertigo with an average remission duration of 3 days. And no recurrence was found in the two patients with cholesteatoma. After a follow-up of 1-6 years, we found a remarkable improvement of air conduction without bone conduction loss in all patients and there was a significant difference between preoperative and post-operative air-bone gap（P<0.05）. Conclusion:With a strict selection according to the indications, MT showes safe and effective long-term outcomes and is proved to be applicable in ossicular chain reconstruction in the cases of fixation of the stapes footplate accompanied with malleus/incus mobility disorder by various causes.

A cationic polymeric prodrug with chemotherapeutic self-sensibilization co-delivering MMP-9 shRNA plasmid for a combined therapy to nasopharyngeal carcinoma.

To obtain a high-efficiency drug and gene co-delivery system to HNE-1 tumor therapy, a polymeric prodrug (PAAs-MTX) with chemotherapeutic sensibilization was synthesized consisting of a GSH-response hyperbranched poly(amido amine) (PAAs) and an antitumor drug of methotrexate (MTX). Then, the targeting molecule to HNE-1 cells, transferrin (Tf), was conjugated to form the Tf-PAAs-MTX. This polymeric prodrug could deliver MMP-9 shRNA plasmid (pMMP-9) again to form the drug and gene co-delivery system of Tf-PAAs-MTX/pMMP-9. The co-delivery system showed the effective drug and gene delivery ability with high cytotoxicity and gene transfection efficiency to HNE-1 cells. Besides that, Tf-PAAs-MTX also showed the chemotherapeutic sensibilization effect, the formulation containing PAAs segments showed much higher cytotoxicity than that of free MTX. Benefiting from the sensibilization effect and MTX/pMMP-9 co-delivery strategy, this Tf-PAAs-MTX/pMMP-9 co-delivery system exhibited the significantly improved therapeutic efficacy to HNE-1 tumor in a combined manner which was confirmed by in vitro and in vivo assays. Moreover, its biocompatibility, especially the blood compatibility was analyzed. This polymeric prodrug provided an easily delivery system combining the drug/gene co-delivery, chemotherapeutic sensibilization and targeting into one single platform, which showed a promising application in nasopharyngeal carcinoma therapy.

Identification of a complex genomic rearrangement in TMPRSS3 by massively parallel sequencing in Chinese cases with prelingual hearing loss.

BACKGROUND: Genetic variants in TMPRSS3 have been causally linked to autosomal recessive nonsyndromic hearing loss (HL) at the DFNB8 and DFNB10 loci. These variants include both single nucleotide and copy number variations (CNVs). In this study, we aim to identify the genetic cause in three Chinese subjects with prelingual profound sensorineural HL. METHODS: We applied targeted genomic enrichment and massively parallel sequencing to screen 110 genes associated with nonsyndromic HL in the three affected subjects. CNVplex® analysis and polymerase chain reaction (PCR) were performed for CNV detection. RESULTS: We identified biallelic variations in TMPRSS3 including a novel complex genomic rearrangement and a novel missense mutation, c.551T>C. We have mapped the breakpoints of the genomic rearrangement and showed that it consisted of two deletions and an inversion encompassing exon 3 to exon 9 of TMPRSS3. CONCLUSION: Our study expanded the mutational spectrum of TMPRSS3 to include complex genomic rearrangements. It showcased the importance of an integrative approach to investigate CNVs and their contribution to HL.

Folate-Targeted pH and Redox Dual Stimulation-Responsive Nanocarrier for Codelivering of Docetaxel and TFPI-2 for Nasopharyngeal Carcinoma Therapy.

Due to the increasing incidence of tumor metastasis and multidrug resistance, even though a combined use of chemotherapy and radiotherapy is introduced, the 5-year average survival rate of an advanced nasopharyngeal carcinoma (NPC) patient still remains low. Hence, targeted slow-release anticancer drugs represent a potential therapy for advanced NPC. In this study, pH and redox dual stimulation-responsive folate-targeted folic acid - ß-cyclodextrin - hyperbranched poly(amido amine)s (FA-DS-PAAs) nanocarriers for codelivery of docetaxel (DOC) and tissue factor pathway inhibitor 2 (TFPI-2) for NPC therapy are discussed. Physical and chemical properties, in vitro DOC-release properties, folic acid (FA)-targeting, transfection, Western blotting, DOC and TFPI-2 codelivery, therapeutic properties, targeted inhibition, and biocompatibility, in vivo FA-targeting, toxicity, and therapeutic properties of FA-DS-PAAs/DOC/TFPI2 nanoparticles are evaluated. The results indicate that the 200 nm low-toxicity FA-DS-PAAs/DOC/TFPI2 nanoparticles could enhance TFPI2 gene expression, make cancer cells more sensitive to DOC, induce cell apoptosis, and reduce cell invasion more effectively compared with monochemotherapy. With respect to the targeted release of drugs (DOC and TFPI2) in tumor cells, FA-DS-PAAs/DOC/TFPI2 is associated with the slowest growth rate of tumor and the smallest volume of tumor, so this study demonstrates the best synergetic antitumor effect. We anticipate that this study is important because it not only provides a potential new therapy approach for NPC but also paves the preclinical way for potential application of FA-DS-PAAs/DOC/TFPI2.

Transferrin-functionalized chitosan-graft-poly(l-lysine) dendrons as a high-efficiency gene delivery carrier for nasopharyngeal carcinoma therapy.

Transferrin-functionalized chitosan-graft-poly(l-lysine) dendrons (CS-PLLD-Tf) were synthesized in this work and then used to deliver the MMP-9 shRNA plasmid to HNE-1 cells for gene therapy. The obtained CS-PLLD-Tf displayed excellent gene transfection in vitro, verifying the strategy of constructing a high-efficiency gene carrier by conjugating the multiple low generation PLLDs to one molecule of CS to form a copolymer. Particularly, CS-PLLD-Tf showed much higher gene transfection efficiency than PEI-25k and CS-PLLD in the presence of serum, and more than 40% HNE-1 cells were transfected. This result led to an obvious decrease in MMP-9 protein expression and the apoptosis of HNE-1 cells as well as inhibition of the growth and invasion of HNE-1 tumors. The in vivo anti-tumor assay showed that CS-PLLD-Tf delivered the MMP-9 plasmid effectively and the tumor volume decreased more than 77% compared with the PBS control group. Moreover, CS-PLLD-Tf displayed higher bioavailability and good biocompatibility such as lower cytotoxicity, excellent blood compatibility and non-toxicity to organs, suggesting a potential application in gene therapy of tumors.

[Analyses of chronic otitis media with intact tympanic membrane concurrent with intracranial complication].

OBJECTIVE: To investigate the clinical and imaging characteristics of chronic otitis media with intact tympanic membrane. METHOD: Ten patients were retrospective studied in the department of otorhinolaryngology of Guangdong general hospital from December 2006 to January 2015. The clinical characteristics of their manifestations, audiology and imaging examinations were analyzed. RESULTS: All the cases could be divided into two groups: the intracranial complication group who was primarily diagnosed as intracranial complications, and the hearing loss group who primarily complained of hearing loss. Five cases belonged to the first group, which include 1 cerebellar abscess, 3 meningitis and 1 meningoencephalitis, 2 of them were profound hearing loss, 2 were mixed hearing loss, and 1 was normal hearing. The other 5 cases belonged to the hearing loss group, 3 were mixed deafness, 2 were conductive deafness. All the case showed positive high-resolution computed tomography (HRCT) features. In the first group, four cases showed the soft tissue shadow in tympanic antrum/superior tympanic pouch and 1 case showed a wide damage of skull base, in addition to intracranial complications. In the second group, five cases showed soft tissue imaging in tympanic antrum/superior tympanic pouch. Pathology showed that 2 cases were cholesterol granuloma(one was in the first group and the other was in the second group), 4 were intracranial inflammatory(the first group) and 4 were cholesteatoma(the seond group). CONCLUSIONS: In chronic otitis media with intact tympanic membrane, intracranial complications may be the primarily diagnosis, which should be paid much attention by multidisciplinary team. HRCT and audiology were valuable for early diagnosis.

[The effect of binaural bi-modal fitting on speech recognition of cochlear implant recipients with low frequency electrodes incompletely implanted].

OBJECTIVE: To study the effect of binaural bi-modal fitting (one cochlear implant and a contralateral hearing aid) on speech recognition of cochlear implant recipients with low frequency electrodes incompletely implanted. METHOD: A total of 15 cochlear implant(CI) users who have low residual hearing (250 Hz 45-90 dBHL, 500 Hz 75 - 100 dBHL) of their non surgery ears. We closed the top three electrodes to simulate the state of low-frequency electrodes incompletely implanted by fitting software. (1) The same tester received twice speech recognition tests in incompletely implanted state, one test used single CI model and the other used binaural bi-modal fitting model. (2)The same tester received above twice speech recognition tests again, but in completely implanted state. Then we compared the results of tests. RESULT: The CI users used binaural bi-modal fitting have higher speech recognition score than the same users used unilateral cochlear implant model in low-frequency electrodes incompletely implanted state(P<0. 05). The gap between the scores of the tests by using two usage patterns in low frequency electrodes incompletely implanted state was more than that in completely implanted state (P<0. 05). CONCLUSION: Binaural bi-modal fitting can help CI users to improve their speech recognition ability in low-frequency electrodes incompletely implanted state.

[The correlations between varying tinnitus severity and anxiety and depression in non-acute tinnitus patients].

OBJECTIVE: To investigate the correlation betwen varying degrees of non-acute tinnitus and anxiety and depression. METHOD: Seventy-seven outpatients with non-acute tinnitus as their in chief complaint were submitted to Tinnitus Handicap Inventory(THI), Self-Rating Anxiety Scale(SAS), and Self-Rating Depression Scale (SDS). RESULT: THI and its three subscales were found to have significant correlations with SAS and SDS. The group (THI ≥ 38) had more anxiety and depression than the mild (THI < 38). Significant correlations were also observed between THI, SAS and SDS in the group with THI ≥ 38, in contrast with the group of THI < 38. CONCLUSION: Patients with THI ≥ 38 suffered from severe anxiety and depression than the mild. Doctors should pay more attention to these patients, especially their psychological disorders.

[Application of HRCT multi-planar reconstruction for measuring cochlear morphology in cochlear implantation].

OBJECTIVE: To explore the value of high-resolution temporal bone CT (HRCT) multi-planar reconstruction (MPR) technique for measuring cochlear morpholog in the preoperative evaluation of cochlear implant. METHOD: We retrospectively analyzed the clinical data of 80 cochlear implant recipients. The groups were investigated in our research according to the patient's sex, age and whether the inner ear is malformed or not. Temporal bone HRCT MPR was applied to analyze the measurement parameter in the standard orientation of the cochlea image: the largest distance from the round window to the lateral wall (distance A), the perpendicular distance (B) and angle of the basal turn of the cochlea relative to the midsagittal plane (γ). Reproducibility of the measured data was evaluated and the results between the different groups were compared. RESULT: Measurement of parameter values between the intraobserver and interobserver showed good reproducibility. In normal group, distance A [(8.79±0.34) mm] and perpendicular distance B [(6.58±0.28) mm] of males' cochleae were significantly larger than A [(8.53±0.47) mm] and B [(6.22±0.43) mm] of females' (P<0.05), but the angle γ was not significantly different between the two groups (P>0.05). The A and B values didn't change with age (P>0.05), but the angle γ tended to decrease with increasing age (r=-0.25, P<0.05). In the malformation group, the value B was significantly smaller than that of the normal group (P<0.01), but for value A and angle γ, there was no significant difference between the two groups (P>0.05). CONCLUSION: HRCT MPR technique is simple and reliable in measurement of cochlear morphology. It can be used to guide the surgery and provide an objective basis for designing the personalized electrode. More clinical application of this technology is recommended.

[The additional 180° roll test in the determination of affected side of horizontal semicircular canal benign paroxysmal positional vertigo].

OBJECTIVE: To evaluate the significance of additional 180-degree roll test (RT) in the determination of affected side in patients with horizontal semicircular canal benign paroxysmal vertigo (HSC-BPPV). METHOD: One hundred and six patients with HSC-BPPV were performed the 90 degree RT. patients whose affected side cannot be determined by 90 degree RT were performed 180 degree roll test. RESULT: The affected side was deter- mined by the 180 degree RT in 10 cases in which the lesion side cannot be determined by the 90-degree RT. CONCLUSION: The affected side of HSC BPPV was able to be determined by 180 degree RT when it not possible to be determined by 90 degree RT. 180 degree RT is an effective and simple additional method.