Refined Sound Therapy in Combination with Cognitive Behavioural Therapy to Treat Tinnitus: A Randomized Controlled Trial.

Background: Post-auricular injection of lidocaine and methylprednisolone sodium succinate is a commonly used treatment for outpatient patients with tinnitus, but it is invasive, painful and has uncertain efficacy. We need to try to replace it with more non-invasive and effective treatments. The 2014 guidelines of the American Academy of Otolaryngology-Head and Neck Surgery recommend the use of cognitive behavioral therapy (CBT) to treat tinnitus. Some clinical doctors have also attempted sound therapy for tinnitus. It is unclear whether sound therapy combined with CBT y is more effective than local injection of lidocaine and methylprednisolone sodium succinate in treating tinnitus. Objective: To evaluate the efficacy and influencing factors of refined sound therapy combined with CBT in the treatment of tinnitus and compare it with post-auricular injection of lidocaine and methylprednisolone sodium succinate. Methods: We recruited 100 patients with tinnitus; ultimately, 81 patients completed the experiment and underwent follow-up. Patients were randomly assigned to either the treatment group (refined sound therapy combined with CBT) or the control group (post-auricular injections of lidocaine and methylprednisolone sodium succinate). Data was collected from 49 patients in the treatment group and 32 patients in the control group. Pre- and post-treatment data were collected using the Self-Rating Depression Scale (SDS), Hamilton Anxiety Rating Scale (HAM-A), Visual Analogue Score (VAS), Tinnitus loudness and Tinnitus Handicap Inventory (THI) score. Comparisons between groups were performed using the chi-square test, Fisher's exact test, or Wilcoxon rank-sum test. All tests were two-sided and considered statistically significant with P < .05. Results: The THI, SDS and HAM-A scores in the treatment group decreased significantly. In the control group, there was a significant reduction in THI scores, but not in SDS and HAM-A scores. In addition, tinnitus loudness and VAS scores were significantly decreased in the 2 groups. There was a significant difference in the reduction of THI, SDS, HAM-A and VAS scores between the 2 groups; the treatment group showed a greater reduction. However, there was no significant difference in the reduction of tinnitus loudness. There was no statistical difference in the reduction of THI scores, SDS scores, VAS scores and tinnitus loudness in different frequency groups, but there was a statistical difference in the reduction of HAM-A scores. There was no statistical difference in the reduction of THI scores, SDS scores, HAM-A scores, VAS scores and tinnitus loudness between patients with and without hearing loss. Conclusions: (1) This new combination is more effective than post-auricular injection of lidocaine and methylprednisolone sodium succinate in treating tinnitus and improving psychological symptoms. The latter had no effect on improving psychological indicators. (2) With this combination, patients with different tinnitus frequencies experienced different improvements in anxiety. (3) Low-frequency tinnitus seems have been more likely to cause sound adaptation. (4) The improvement in tinnitus and anxiety was the same regardless of whether or not there was hearing loss.

BIX-01294, a G9a inhibitor, suppresses cell proliferation by inhibiting autophagic flux in nasopharyngeal carcinoma cells.

G9a, a histone methyltransferase, has been found to be upregulated in a range of tumor tissues, and contributes to tumor growth and metastasis. However, the impact of G9a inhibition as a potential therapeutic target in nasopharyngeal carcinoma (NPC) is unclear. In the present study we aimed to investigate the anti-proliferative effect of G9a inhibition in the NPC cell lines CNE1 and CNE2, and to further elucidate the molecular mechanisms underlying these effects. The expression of G9a in NPC tumor tissues was significantly higher than that in normal nasopharyngeal tissues. The pharmacological inhibition of G9a by BIX-01294 (BIX) inhibited proliferation and induced caspase-independent apoptosis in NPC cells in vitro. Treatment with BIX induced autophagosome accumulation, which exacerbated the cytotoxic activity of BIX in NPC cells. Mechanistic studies have found that BIX impairs autophagosomes by initiating autophagy in a Beclin-1-independent way, and impairs autophagic degradation by inhibiting lysosomal cathepsin D activation, leading to lysosomal dysfunction. BIX was able to suppress tumor growth, possibly by inhibiting autophagic flux; it might therefore constitute a promising candidate for NPC therapy.

BIX-01294-enhanced chemosensitivity in nasopharyngeal carcinoma depends on autophagy-induced pyroptosis.

Nasopharyngeal carcinoma (NPC) is a common cancer in southern China and Southeast Asia. Nowadays, radiotherapy is the therapy of choice for NPC patients, and chemotherapy has been found as an alternative treatment for advanced NPC patients. However, finding novel drugs and pharmacologically therapeutic targets for NPC patients is still urgent and beneficial. Our study showed that BIX-01294 (BIX) can induce autophagic vacuoles formation and conversion of LC3B-I to LC3B-II in NPC cells in both dose- and time-dependent manners. Notably, the combination of BIX and chemotherapeutic drugs significantly decreased the cell viability and increased the lactate dehydrogenase release. Meanwhile, BIX plus cis-platinum (Cis) treatment induced pyroptosis in NPC cells as featured by cell swelling and bubble blowing from the plasma membrane, the increased frequency of annexin V and propidium iodide (PI) double-positive cells, as well as the cleavage of gasdermin E (GSDME) and caspase-3. Moreover, the deficiency of GSDME completely shifted pyroptosis to apoptosis. Furthermore, the inhibition of autophagy by chloroquine and the knockout of ATG5 gene significantly blocked the BIX-induced autophagy as well as pyroptosis in both in vitro and in vivo studies. Our data demonstrated that BIX-combined chemotherapeutic drugs could induce the Bax/caspase-3/GSDME-mediated pyroptosis through the activation of autophagy to enhance the chemosensitivity in NPC.

An epidemiological survey of laryngopharyngeal reflux disease at the otorhinolaryngology-head and neck surgery clinics in China.

PURPOSE: Using the Reflux Symptom Index (RSI), this nationwide study aimed to investigate the incidence, diagnostic status, risk factors, and common symptoms of adult laryngopharyngeal reflux disease (LPRD) at otorhinolaryngology-head and neck surgery (OHNS) clinics in China. METHODS: This multicenter cross-sectional survey began at the different institutions ranged from July to October 2017, and the duration was 12 months. A total of 90,440 eligible patients were finally enrolled from 72 medical institutions in China. All these patients completed the questionnaire based on RSI. In this study, LPRD was defined as RSI > 13. RESULTS: There were 9182 with LPRD among the 90,440 eligible participants (10.15%). However, only 1294 had a history of LPRD diagnosis among those with LPRD (14.09%). There were regional differences in the frequency of LPRD (P < 0.001). The proportions of patients with LPRD in males (vs. females), middle- and old-aged patients (vs. young), with current smoking history (vs. no smoking), and current drinking history (vs. no drinking) were significantly higher (all P < 0.001). Middle and old age, current smoking, and drinking history were independent predictors of LPRD (all P < 0.001, OR 1.240, 1.261, and 1.481, respectively). "Sensations of something stuck in throat or a lump in throat", "clearing throat", and "excess throat mucus or postnasal drip" were the most frequent clinical symptoms in patients with LPRD. CONCLUSIONS: LPRD has a high incidence at the OHNS clinics in China. However, the diagnostic status of this disease is not optimistic. Older age, smoking, and drinking history were risk factors for LPRD.

Confirmed laryngeal sarcomatoid carcinoma after three vocal cord surgeries in a short period of time: A case report and literature review.

Sarcomatoid carcinoma is a rare disease, while larynx sarcomatoid carcinoma is a rarer type of cancer characterized by high malignancy and rapid growth. This type of cancer is prone to misdiagnosis and recurrence. The present case study aimed to provide a detailed report on the diagnosis and treatment of a male patient who underwent three tumor resections in the same location of the vocal cords within a short time. Immunohistochemical analysis was not performed after the first two surgeries at other hospitals, thus resulting in an inconclusive diagnosis. Within a short time, it was suspected that the patient experienced local recurrence, thus rapidly leading to the onset of grade III laryngeal obstruction symptoms. During his last visit to the Second Affiliated Hospital of the Army Military Medical University Outpatient Department (Chongqing, China), a biopsy was carried out and tissue samples were collected for immunohistochemical examination. The diagnosis of larynx sarcomatoid carcinoma was therefore verified. Subsequently, partial laryngectomy combined with laryngeal functional reconstruction and left neck lymph node dissection were performed. Post-surgery immunohistochemical analysis also verified larynx sarcomatoid carcinoma. During the 6-month follow-up, there was no evidence of metastasis or recurrence.

Metformin attenuates cadmium-induced degeneration of spiral ganglion neuron via restoring autophagic flux in primary culture.

Cadmium (Cd), a common environmental and occupational toxicant, is an important risk factor for hearing loss. After exposure, Cd accumulates in the inner ear and induces spiral ganglion neuron (SGN) degeneration; however, the underlying mechanisms are poorly understood. Dysfunctional autophagy has been implicated in many neurodegenerative diseases, including Cd-induced neurotoxicity. Metformin has been validated to confer not only anti-hyperglycaemic but also neuroprotective effects. However, the relationship between autophagy dysfunction, SGN degeneration, and the effect of metformin on Cd-induced SGN neurotoxicity has not yet been established. In this study, we demonstrate that metformin notably attenuates Cd-evoked SGN degeneration by restoring impaired autophagy flux, as evidenced by the suppression of Cd-induced elevation of autophagy markers microtubule-associated protein 1A/1B-light chain 3-II (LC3-II) and autophagy substrate protein p62 in degenerated SGN. Blockage of autophagy flux by chloroquine abolished metformin-induced neuroprotection against Cd-induced neurotoxicity in SGN. The results of this study reveal that autophagy dysfunction is an important component of Cd-induced SGN degeneration, and metformin may be a potential protective agent for attenuating SGN degeneration following Cd exposure.

New anatomic division of the orbital and sinus regions: guidance for nasal endoscopic resection of space-occupying lesions.

The significance of anatomic divisions of the orbital and sinus regions in providing guidance for nasal endoscopic resection of benign space-occupying lesions was discussed. This retrospective study included 177 cases with benign space-occupying lesions of the orbital and sinus resected between 2001 and 2009. Locations of lesions were assigned to 3 anatomic categories. The 63 cases of benign lesions of the sphenoethmoid-orbital region underwent nasal endoscopic resection. In the 31 cases of benign lesions in the frontoethmoid-orbital region, surgical approaches included nasal endoscopy (n = 15) and combined endoscopic and external (n = 9) and external approaches (n = 4). In 87 cases of lesions in the axillae-ethmoid-orbital region, surgical approaches included nasal endoscopy (n = 67), combined approaches (n = 15), and external approaches (n = 5). Endoscopy facilitated the resection of both 63 cases of benign lesions of sphenoethmoid-orbital region, and 82 cases of benign lesions of the frontoethmoid-orbital and maxillae-ethmoid-orbital regions with good prognosis. However, the procedure was relatively more difficult for the other 32 cases of benign lesions of the frontoethmoid-orbital or maxillae-ethmoid-orbital regions. The feasibility of nasal endoscopic resection differs markedly according to anatomic location. Preoperative classification of the site of the lesions will help to define the indications for nasal endoscopic resection of the orbital and sinus regions.

Expression of G9a in Auditory Cortex Is Downregulated in a Rat Model of Age-Related Hearing Loss.

G9a is essential for dendritic plasticity and is associated with neurological disorders. The possible relationship between age-related hearing loss and G9a expression in the auditory cortex has not been fully explored. This study aimed to understand the expression patterns of G9a-mediated histone methylations in the auditory cortex during aging. Using immunofluorescence and western blotting, we demonstrated that a significant reduction in G9a expression observed in the auditory cortex of 24-month-old rats compared to 3-month-old rats, was associated with remarkable hearing threshold elevation and hair cell loss. Correspondingly, histone H3 lysine 9 (H3K9) mono- and dimethylation (marked by H3K9me1 and H3K9me2, respectively), which were regulated by G9a activity, also evidently decreased during aging. These findings, which merit further investigation, suggest a possible association between G9a-mediated histone methylations and central age-related hearing disorders.

[Analysis of the association between helicobacter pylori infection and laryngeal lesions].

Objective:To investigate the relationship between Helicobacter pylori（H.pylori） infection and laryngeal lesions. Method:204 patients with laryngeal lesions were arranged into laryngeal lesions group, and 150 healthy persons who were willing to accept the electronic fiber gastroscopy examination in the same period were selected as control group. The positive rate of H.pylori infection in the two groups was observed and the association between H.pylori infection status and clinical characteristics in patients with laryngeal diseases was analyzed. Result:The positive rate of H.pylori infection in laryngeal lesion group and control group were 56.86% and 47.33%（χ²=3.150, P=0.076）, respectively. Among the patients with laryngeal lesion, the positive detection rate of H.pylori was not associated with age, gender, or gastric disease history. The positive rate of H.pylori infection in benign lesions, precancerous lesions and laryngeal malignant lesions were 53.70%, 55.56% and 75.00%, respectively. The difference of positive rate of H.pylori infection in laryngeal malignant lesions was significantly higher than other kinds of lesions（χ²=6.338, P=0.012）. Among laryngeal precancerous lesions and laryngeal malignant lesions patients, the appearance rate of gastrointestinal lesions were significantly higher in the patients with positive H.pylori infection than those without H.pylori infection（P<0.05）. Conclusion:H.pylori infection was positively related to the severity of laryngeal lesions and highly positively related to laryngeal malignant lesions.

Identification of a novel mutation in CRYM in a Chinese family with hearing loss using whole-exome sequencing.

Previous studies have identified ~50 genes that contribute to non-syndromic autosomal dominant sensorineural deafness (DFNA). However, in numerous families with hearing loss, the specific gene mutation remains to be identified. In the present study, the clinical characteristics and gene mutations were analyzed in a Chinese pedigree with hereditary hearing loss. The clinical characteristics of the family members were assessed and a detailed audiology function examination was performed. Whole-exome sequencing (WES) was performed to identify the gene mutation responsible for the hearing loss. Sanger sequencing was used to verify the candidate mutation detected in the family. The family consisted of 31 members, seven of whom were diagnosed with sensorineural deafness of varying degrees. No mutation was identified by the general deafness gene chip. However, a novel heterozygous mutation in exon 3 (c.152C>T; Pro51Leu) of the gene crystallin µ (CRYM) was identified by WES. This result was further verified by Sanger sequencing. Co-segregation of genotypes and phenotypes suggested that this novel mutation was instrumental for the hearing loss/DFNA. In conclusion, the present study identified a novel pathogenic mutation, NM_001888.5(CRYM): c.152C>T(Pro51Leu), associated with DFNA. This mutation has not been reported previously and further functional studies are warranted.

Scopolamine attenuates auditory cortex response.

CONCLUSION: Scopolamine, a tropane alkaloid drug that mainly acts as an antagonist of muscarinic acetylcholine receptors, was found to reduce the local field potentials (LFP) of auditory cortex (AC) evoked by tone and gap-offsets whose effects may compensate the cortical hyperexcitability related to tinnitus. OBJECTIVE: To study the effects of scopolamine on the AC and the inferior colliculus (IC) of awake rats in order to understand scopolamine's effect on tinnitus and gap detection. METHOD: Silent gaps (duration varied from 2-100 ms) embedded in otherwise continuous noise were used to elicit AC and IC response. Gap evoked AC and IC field potentials were recorded from awake rats before and after treatment of scopolamine (3 mg/kg, i.m.). RESULTS: Acute injection of scopolamine (3 mg/kg, i.m.) induced a significant reduction of the AC response, but not the IC response, to the offset of the gaps embedded in white noise. The results suggest that scopolamine may reduce AC neural synchrony.

Noise exposure enhances auditory cortex responses related to hyperacusis behavior.

Hyperacusis, a marked intolerance to normal environmental sound, is a common symptom in patients with tinnitus, Williams syndrome, autism, and other neurologic diseases. It has been suggested that an imbalance of excitation and inhibition in the central auditory system (CAS) may play an important role in hyperacusis. Recent studies found that noise exposure, one of the most common causes of hearing loss and tinnitus, can increase the auditory cortex (AC) response, presumably by increasing the gain of the AC. However, it is not clear whether the increased cortical response will affect sound sensitivity and induce hyperacusis. In this experiment, we studied the effects of noise exposure (narrow band noise, 12 kHz, 120 dB SPL, 1 hour) on the physiological response of the inferior colliculus (IC) and the AC, and the behavioral sound reaction in conscious Sprague Dawley rats. Noise exposure induced a decrease of sound evoked potential in the IC. However, significant increases of AC response including sound evoked potentials and the spike firing rates of AC neurons were recorded right after the noise exposure. These results suggest that noise exposure induces hyperexcitability of AC presumably by increasing the post-synaptic response of AC neurons. The behavioral consequence of the noise exposure on sound perception was measured by the amplitude of the acoustic startle response before and after noise exposure in a separate group of rats. Although noise exposure caused a moderate hearing loss, the acoustic startle amplitude at the super-threshold level was significantly increased. These results suggest that noise exposure can cause exaggerated the sound reaction which may be related with the enhanced responsiveness of the AC neurons. This phenomenon may be related with noise induced hyperacusis.This article is part of a Special Issue entitled: Tinnitus Neuroscience.

[Histological changes of peripheral vestibular organs in the inner ears of Smad4 conditional knockout mice].

OBJECTIVE: To investigate the histological changes in the vestibular endorgans of Smad4 gene conditional knockout mice and to explore the influence of the Smad4 gene on vestibular development. METHODS: Histological changes of periphery vestibular organs in inner ear of Smad4 conditional knockout mice were investigated by frozen sections, immunofluorescence, confocal microscopy, scanning electron microscopy and transmission electron microscopy. RESULTS: There was no Smad4 expression in the inner ear cartilage capsule of Smad4-/- mice. In Smad4+/- mice, Smad4 expression in the same cartilage capsule was positive, and it was strong positive in Smad4+/+ mice. Smad4 expression in vestibular sense epithelium, crista ampullaris and macula, was positive. And no difference was found among these three genotypes. Studying at scanning electron microscopy and transmission electron microscopy levels and anti-filament immunofluorescence showed that no pathological changes were observed in all the three genotype mice. CONCLUSION: Although the Smad4 gene was knockout effectively in the auricular cartilage capsule of Smad4 conditional knockout mice,the histological changes of Smad4 conditional knockout mice in vestibulum auris internal were slightly.

The role of Smad4 in vestibular development in mice.

The regulation of the bone morphogenetic protein (BMP) signal transduction pathway is important in the development of the inner ear and vestibular system. We reported previously that small mothers against decapentaplegic homolog-4 (Smad4) is required for inner ear cochlear development and normal auditory function in mammals; however, the distribution and functional mechanisms of Smad4 at various stages of vestibular development remained unclear. To investigate the relationship between the Smad4 gene and vestibular organ development, we measured changes in the expression of BMP4 and Smad4 during vestibular development in C57BL/6 mice. In addition, vestibular structures, pathologic changes, and the vestibular function of chondrocyte-specific Smad4 knockout mice were compared to those of the control group. We found that the expression of Smad4 in the inner ear was delayed compared with that of BMP4. Moreover, chondrocyte-specific Smad4 knockout homozygous mice showed stunted growth and partial vestibular deformities, but it showed less histologic changes in the vestibular end-organs and saccule dysfunction. These results suggest that Smad4 participates in late-stage shaping of the configuration of the vestibule and development of vestibular functional, but a Smad4-independent pathway for the inner ear vestibular BMP4 signal transduction could not be rule out.

Temporal processing in inferior colliculus and auditory cortex affected by high doses of salicylate.

Speech recognition and language learning can be affected by both peripheral and central auditory system impairment. However, whether sensorineural hearing loss would affect central auditory processing is not clear. Recent studies found that salicylate not only affects outer hair cell motility in the cochlea, but also blocks GABAergic neuron activities in central nervous systems. This provides a good animal model to evaluate the role of sensorineural hearing loss and central inhibition in auditory temporal processing. In this study, gap prepulse inhibition (gap-PPI) of the acoustic startle reflex was used to measure effects of salicylate on gap detection acuity. Salicylate administration (250 mg/kg) resulted in a significant reduction in gap-PPI amplitude and an increased gap detection threshold at 50 dB SPL, but not at 60 or 80 dB SPL. To identify the physiological effects of salicylate on central auditory system function, the inferior colliculus (IC) and auditory cortex (AC) responses were measured from conscious rats with chronically implanted electrodes. Salicylate induced a significant increase of the gap-detection threshold in AC-evoked potentials, but not in the IC-evoked potentials. The AC gap-detection threshold shift was diminished measured at an equal sensational level. These results suggest that salicylate-induced temporal processing deficits may be due to peripheral hearing loss, not central disinhibition.

[Distributing and changes of NO and calcitonin gene-related peptide in pharyngeal tissues and plasma in obstructive sleep apnea and hypopnea syndrome].

OBJECTIVE: To explore the distributing and changes of nitric oxide (NO) and calcitonin gene-related peptide (CGRP) in obstructive sleep apnea and hypopnea syndrome (OSAHS). METHODS: This study investigated the content of NO and CGRP in plasma and in pharyngeal tissues of OSAHS subjects. Moreover, the distribution of NOS and CGRP in pharyngeal tissues was researched using method of immunohistochemistry. RESULTS: (1) Plasma NO level in control group was higher than in mild OSAHS group and in serious OSAHS group, P < 0.001. There was a significant increase in plasma CGRP after UPPP (P < 0.001), and the plasma CGRP level in control group was as high as that in after UPPP group (P > 0.05) NO level in pharyngeal tissue of OSAHS increased significantly, P < 0.001), but CGRP level decreased markedly, P < 0.001. (2) Immunohistochemistry change about NOS: there was a feeble positive expression in mucosa squamous epithelium of OSAHS and powerful positive expression in some salivary glandular epithelium, striated muscle cells, vascular wall/small vascular wall and epithelium of glandular duct of OSAHS. (3) Immunohistochemistry change about CGRP: There were diffusible positive markings in striated muscle cells and most cilium columnar epithelium in OSAHS, and negative expressions in other sites. CONCLUSIONS: (1) Circulating NO and CGRP are suppressed in OSAHS, and reversible promptly after UPPP. Further more, The content of NO in pharyngeal tissues increases significantly and that of CGRP decreases markedly. (2) The distribution of NOS and CGRP in pharyngeal tissue changes apparently according to sites.