A pharynx-to-brain axis controls pharyngeal inflammation-induced anxiety.

Anxiety is a remarkably common condition among patients with pharyngitis, but the relationship between these disorders has received little research attention, and the underlying neural mechanisms remain unknown. Here, we show that the densely innervated pharynx transmits signals induced by pharyngeal inflammation to glossopharyngeal and vagal sensory neurons of the nodose/jugular/petrosal (NJP) superganglia in mice. Specifically, the NJP superganglia project to norepinephrinergic neurons in the nucleus of the solitary tract (NTSNE). These NTSNE neurons project to the ventral bed nucleus of the stria terminalis (vBNST) that induces anxiety-like behaviors in a murine model of pharyngeal inflammation. Inhibiting this pharynx→NJP→NTSNE→vBNST circuit can alleviate anxiety-like behaviors associated with pharyngeal inflammation. This study thus defines a pharynx-to-brain axis that mechanistically links pharyngeal inflammation and emotional response.

Incidence and characteristics of otitis media with effusion in adults before, during, and after the COVID-19 pandemic.

OBJECTIVES: To investigate the incidence and characteristics of adult otitis media with effusion (OME) before, during, and after the COVID-19 pandemic. METHODS: A retrospective descriptive study was conducted. The incidence, age, sex, affected ear side, time of OME onset according to COVID-19 and days of improvement after conservative treatment were determined to assess the clinical features of adult OME in different periods of the COVID-19 pandemic. RESULTS: The incidence of adult OME during these periods was 3.17%, 2.30%, 6.18%, and 3.68%, respectively. Unilateral ear involvement and male sex were more common. The onset of adult OME occurred 7.80 ± 3.97 days after COVID-19 diagnosis, and improvement was observed after 12.24 ± 5.08 days of conservative treatment. Patients in the post-pandemic period were older than those in the non-pandemic period. CONCLUSION: The incidence of adult OME in China showed a tendency to decrease, recover, and decrease again following the COVID-19 outbreak. Pandemic prevention and control measures have had a certain impact on reducing the incidence, but the elderly are more prone to this disease.

Increased expression of ST2 on regulatory T cells is associated with cancer associated fibroblast-derived IL-33 in laryngeal cancer.

Interleukin (IL)- 33 plays an essential role in regulatory T cell (Treg)-mediated immunosuppression in cancers and underlies the crosstalk between Tregs and the tumor microenvironment. However, the phenotypic characteristics of subset Tregs modulated by IL-33 and its association with the tumor microenvironment are not fully understood. This study aimed to examine the expression of ST2, the receptor of IL-33, on Tregs in tumors and to evaluate their association with cancer associated fibroblasts (CAFs) and reciprocal influences on the prognosis of laryngeal cancer. Our results showed that increased numbers of Tregs were found in laryngeal tumor tissues. Tregs in stromal IL-33-positive tumor tissues demonstrated significantly higher expression of ST2 than those in IL-33- or adjacent nontumor tissues. ST2-expressing Tregs exhibited upregulation of Ki67 and CTLA4 compared with their ST2- negative counterparts. Furthermore, IL-33 in the tumor microenvironment was mainly derived from fibroblasts. ST2 expression on Tregs was correlated with the number of IL-33-positive CAFs. High ST2 expression on Tregs, combined high ST2 on Tregs and the presence of IL-33 expressing CAFs was associated with worse survival outcomes in laryngeal cancer. This study indicated that increased expression of ST2 on Tregs is associated with microenvironmental IL-33 signaling derived from CAFs in laryngeal cancer, unraveling the special role of Tregs and fibroblasts in modulating IL-33/ST2 involved immune-evasive tumor microenvironment.

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.

lncRNA SOX2-OT regulates laryngeal cancer cell proliferation, migration and invasion and induces apoptosis by suppressing miR-654.

Laryngeal carcinoma is the most common type of malignant tumor in the head and neck. Long non-coding RNAs (lncRNAs) serve crucial roles in numerous biological processes. The present study aimed to investigate the role of lncRNA SOX2-OT in laryngeal cancer and to reveal the underlying mechanisms. Reverse transcription-quantitative PCR assays were used to measure the expression levels of SOX2-OT in the laryngeal cell lines. Furthermore, cell proliferation, apoptosis, migration and invasion were assessed by CCK-8, flow cytometry, wound healing and Transwell assays, respectively. Western blot assay was performed to detect the protein expressions. In addition, a dual-luciferase reporter assay was performed to confirm the direct interaction between SOX2-OT and microRNA (miR)-654. The data demonstrated that SOX2-OT level were significantly increased in the laryngeal cell lines. Furthermore, SOX2-OT silencing markedly promoted apoptosis and suppressed the proliferation, migration and invasion of TU-177 cells. A dual-luciferase reporter assay revealed that miR-654 was a direct target of SOX2-OT. Moreover, downregulation of miR-654 could attenuate cell apoptosis and accelerate cell proliferation, migration and invasion in TU-177 cells. In summary, the present study reported that knockdown of SOX2-OT could suppress cell proliferation, migration and invasion, and induce apoptosis in laryngeal cancer by targeting miR-654.

BMI1'S maintenance of the proliferative capacity of laryngeal cancer stem cells.

BACKGROUND: In laryngeal squamous cell carcinoma (SCC), CD133+ cells were found to display cancer stem cell (CSC) characteristics. BMI1 is an oncogene that plays key roles in proliferation in CSCs. However, no published reports have examined the role of BMI1 in laryngeal CSCs. METHODS: Immunofluorescence staining confirmed the coexpression of BMI1 and CD133. After sorting, real-time polymerase chain reaction (PCR) revealed BMI1 was differentially expressed in CD133+ cells. BMI1 was knocked down and proliferation, colony formation, and apoptosis assays were performed. The influence on CD133+ cells was determined by flow cytometry, sphere-formation assay, and quantitative PCR. Tumorigenicity assays were performed, and the impact on related genes was evaluated. RESULTS: BMI1 was highly enriched in CD133+ cells. BMI1 maintained CD133+ cell proliferation and prevented apoptosis. Gene expression analysis suggested BMI1 regulated alternate cellular pathways. CONCLUSION: BMI1 was required to maintain the proliferative capacity of laryngeal CSCs, which may be a molecular target to cure patients with laryngeal SCC.

BMI1 promotes the progression of laryngeal squamous cell carcinoma.

BMI1 is highly expressed in several malignant tumors, and its expression level is associated with tumor progression, proliferation, and prognosis. However, no published studies have examined the role of BMI1 in laryngeal squamous cell carcinoma (SCC). Expression of BMI1 in primary tumors was analyzed by immunofluorescence staining, real-time PCR, and Western blotting. BMI1 was knocked down, and proliferation, apoptosis, and cell cycle assays were performed. Sensitivity to radiochemotherapy was evaluated, and tumorigenicity assays were performed in vivo. BMI1 was highly expressed in laryngeal SCCs. BMI1 promoted cell proliferation and tumor progression, and inhibited apoptosis due to influences on the cell cycle. More importantly, BMI1 suppressed the sensitization of laryngeal Hep2 cells to radiochemotherapy. BMI1 is essential to maintain the proliferation and progression of laryngeal SCCs. Therefore, depletion of BMI1 may be a potential therapeutic option for cancer management.

Characterization of side population cells from laryngeal cancer cell lines.

BACKGROUND: Analysis of the hematopoietic system has shown that bone marrow stem cells contain a subpopulation that effluxes the DNA binding dye, Hoechst 33342, out of the cell membrane. These cells are called side population (SP) cells and have been shown to have stem cell characteristics. SP cells have already been found in solid tumors, although there are no published reports examining the role of SP cells in human laryngeal cancer. METHODS: SP cells were identified in the laryngeal cancer cell lines, AMC-HN-8 (head and neck cancer cell line 8, from the Asan Medical Center) and Hep-2 (human epithelial type 2) cell lines, by flow cytometry. Hep-2 SP cells were isolated and analyzed in proliferation assays, differentiation studies, radiotherapy resistance studies, and tumorigenesis ability. RESULTS: The AMC-HN-8 and Hep-2 cell lines contained 11.8 ± 1.7% and 17.1 ± 2.0% SP cells, respectively. SP cell proportion was reduced by verapamil. Hep-2 SP cells exhibited higher self-regeneration, proliferation, radiotherapy resistance, and tumorigenicity. CONCLUSIONS: SP cells purified from the Hep-2 cell line harbor cancer stem cell-like properties, and they are heterogeneous, indicating that SP cells are not identical to stem cells.

Canal wall-down mastoidectomy with mastoid obliteration for pediatric cholesteatoma.

CONCLUSIONS: As cholesteatoma is widespread and erosive, it should be cleared away thoroughly, to keep a dry ear and conserve maximum hearing ability in children. Canal wall-down mastoidectomy with mastoid and epitympanum obliteration is a good choice for treatment of cholesteatoma in children. OBJECTIVE: To investigate canal wall-down mastoidectomy with mastoid and epitympanum obliteration for the treatment of cholesteatoma in children. PATIENTS AND METHODS: A retrospective analysis of all cases of pediatric middle ear cholesteatoma in children aged 5-12 years between 1999 and 2006 was conducted in Anhui Provincial Hospital. The follow-up information was completed. Forty-five patients (48 ears) were treated with canal wall-down mastoidectomy with mastoid and epitympanum obliteration and followed up for 2-5 years (mean 3.1 years). RESULTS: The recurrence rate and residual rate were 4.16% and 0%, respectively. All ears were dry within 8-10 weeks. Aural discharge was detected in two cases and controlled with antibiotic ear drops without complications. The dry ear rate was 95.8%. The air-bone gap closure had a mean of 17.2 +/- 2 dB. No obvious complication occurred after operations.

[Prevention of anterior glottic stenosis after CO2 laser cordectomy with chitosan].

OBJECTIVE: To observe the effect of immediate topical application of chitosan on preventing anterior glottic stenosis (AGS) after microsurgical resection of both vocal fold with CO2 laser, including the anterior commissure, in a canine model. METHODS: Sixteen canine larynges were injured by microresecting procedure of both vocal folds with CO2 laser. The dogs were randomly divided into two groups, chitosan group and control group. The chitosan and isotonic sodium chloride solution (control) were used for 5 minutes immediately after surgery. One week after the initial surgery, three dogs in each group were randomly selected , ultrastructure of fibroblast were examined with transmission electronic microscope and expression of basic fibroblast growth factor (bFGF) and transforming growth factor beta1 (TGF-beta1) were evaluated by enzyme-linked immunosorbent assay (ELISA). Three weeks after surgery, the rest dogs' glottic web were lysed and repeatedly treated with chitosan and isotonic sodium chloride solution respectively. The glottic wound healing and AGS formation were examined every week, and all larynges were harvested and examined histologically six weeks after the initial surgery. RESULTS: Transmission electronic microscope examination of the ultrastructure of fibroblast indicated that chitosan inhibited the proliferation of fibroblast. Chitosan increased the expression of bFGF and TGF-beta1, and bFGF and TGF-beta1 in chitosan group, which was significantly higher than that in control group (z=-2.887 and -2.005, P=0.002 and 0.041). Chitosan decreased the extent of AGS formation. Three weeks after the surgery, the AGS lesion in the control group affected mean 49% of the length of the vocal folds from the anterior commissure to the vocal process, while chitosan group affected mean 7%, which was significantly less than the extent of web formation in the control group, (z=-2.619, P=0.008). The grade of collagen content in chitosan group was significantly lower than that in control group (P=0.003). CONCLUSION: Chitosan is effective in preventing AGS after CO2 laser cordectomy.

Peri- and post-operative complications after carbon dioxide laser surgery of the larynx.

OBJECTIVE: To analyze the complications of transoral laser surgery (TLS) for larynx tumors in a single center institution, and to highlight its possible risks. METHODS: This is a retrospective study of the peri- and post-operative complications of 500 patients who underwent TLS from November 2001 to July 2008 in the Department of Otolaryngology, Head and Neck Surgery, Anhui Provincial Hospital, Anhui Medical University, Hefei, China. RESULTS: The total rate of complications was 10.2%. The major complications accounted for 0.8%, including one ignition, one post-operative bleeding, one dyspnea, and one laryngeal web; and the minor ones were found in 47 of 500 patients (9.4%), including 30 patients with suspension laryngoscopy related complications. A higher incidence of complications was observed in patients with larynx carcinoma than those with benign/precancerous lesions (p=0.00). CONCLUSION: Peri- and post-operative complications after TLS for benign larynx tumors are relatively rare events, but are relatively frequent for larynx cancers. Every surgeon who performs laser surgery should have a clear concept of management for these complications, and strategies to avoid these complications should be performed.

[Transoral endoscopic CO2 laser surgery for hypopharynx in experimental animals].

OBJECTIVE: To explore the substantial resection limits of CO2 laser surgery for hypopharynx and the course of wound healing in animals, for the purpose of evaluating the clinic usefulness of transoral CO2 laser surgery in the treatment of selected hypopharyngeal carcinomas. METHODS: Twenty-three dogs were randomly assigned to two groups. Group one (11 dogs) received left piriform sinus resection, group two (12 dogs) received the resection of posterior wall of the hypopharynx. Six dogs in group one were killed immediately or 4, 8, 12, 16, 20 d post-operatively. Seven dogs in group two were killed immediately or 7, 14, 21, 28, 35, 42 d post-operatively. The whole larynx and hypopharynx were taken out and the specimens were examined by naked eyes and under microscope. The other 5 dogs in each group were fed until the wound healed, the duration were observed. RESULTS: All the operations were successful and the results were satisfactory. In group one, the dogs could take food the day after operation; two dogs had slight cough during eating and recovered after five days. In group two, the dogs could take food the next day after operation, eight dogs had slight cough during eating and recovered after ten days. The excision dimension was satisfactory. In group one (resection of the lateral wall of piriform sinus), the size of raw surface was (7.5 +/- 0.8) cm2 (x +/- s) and the healing time was (18.4 +/- 1.5) d. In group two (resection of the posterior wall of the hypopharynx), the wound surface was (7.0 +/- 0.5) cm2 and the healing time was (39.8 +/- 1.9) d. The healing time in group two was significantly longer than that in group one (t = 19.535, P <0. 01). The post-operative healing process were observed, including cellulose membrane coverage, granulation filling and epithelization. CONCLUSIONS: Transoral CO2 laser was suitable for partial hypopharynx resection. Animals can recuperate well with little complications. Although the course of wound healing was delayed, wound surface can recover with good laryngeal and deglutition functions.

Comparison between mitomycin C and chitosan for prevention of anterior glottic steno after CO2 laser cordectomy in dogs.

OBJECTIVE: To compare the effectiveness of an immediate application of topical mitomycin C (MMC) with chitosan for prevention of anterior glottic stenosis (AGS) after microsurgical resecting of both vocal fold with CO2 laser, including the anterior commissure, in a canine model. DESIGN: Prospective randomized experimental study. METHOD: Twenty-four canine larynges were injured by a microresecting procedure of both vocal folds with a CO2 laser. The dogs were randomly divided into three groups for treatment, respectively with MMC, chitosan, or isotonic sodium chloride solution (control) for 5 minutes immediately after surgery. One week after the initial surgery, three dogs in each group were randomly selected, the ultrastructures of the fibroblasts were examined with a transmission electronic microscope, and expression of basic fibroblast growth factor (bFGF) and transforming growth factor beta1 (TGF-beta1) were evaluated by enzyme-linked immunosorbent assay (ELISA). Three weeks after surgery, the remaining dogs' glottic web were lysed and repeatedly treated with MMC, chitosan, or isotonic sodium chloride solution. The glottic wound healing and AGS formation were examined every week, and all larynges were harvested and examined histologically 6 weeks after the initial surgery. RESULTS: Results included 1) transmission electronic microscope examination of the ultrastructures of the fibroblasts indicated that both MMC and chitosan inhibited the proliferation of fibroblast; 2) the expression of bFGF and TGF-beta1 increased both in the MMC group and the chitosan group, and the expression of bFGF in the chi-tosan group was higher than in the MMC group (P = .010), while the expression of TGF-beta1 was lower (P = .000);3) the extent of AGS formation was decreased both in the MMC group and the chitosan group, while it was less in the chitosan group than in the MMC group (P = .047); 4) the grade of collagen content between the MMC group and the chitosan group was not significantly different (P = .5). CONCLUSION: Chitosan is more effective in preventing AGS after CO2 laser cordectomy than MMC.

Influence of carbon dioxide laser irradiation on the morphology and function of guinea pig cochlea.

Both experimental and clinical studies have demonstrated that carbon dioxide laser is suitable for stapedotomy. The aim of this study was to investigate morphological, electrophysiological and functional changes in the inner ear after irradiation with CO(2) laser set with different energy parameters. A cochleostomy in the basal cochlear turn of guinea pig cochleae was performed with CO(2) laser of 1, 2 and 3 w, respectively. The cochleae were removed three weeks after laser irradiation. The auditory evoked brainstem response (ABR) was measured before and after laser application and immediately before removal of the cochlea. Immunohistochemical methods were used to examine inducible nitric oxide synthase (iNOS/NOSII) and heat-shock protein 70 (Hsp70) concentrations in the cochlea after laser application. The organ of Corti was studied by scanning electron microscopy. Worse hearing loss was observed in animals receiving higher-power CO(2) laser. These findings correlated with more intense injury of the cochlear ultrastructure and with positive expression of iNOS and Hsp70 in spiral ganglion cells, nerve fibres, supporting cells of the organ of Corti and cells of the spiral ligament. The CO(2) laser as a noncontact procedure is shown to be effective and safe if the total amount of energy is kept within the limits applied in this study. Nitric oxide and stress proteins play important roles in the traumatic mechanism of the inner ear, which are related to hearing loss and injury of the ultrastructure of the inner ear.