Risk factors and healing factors for pharyngocutaneous fistula after total laryngectomy for laryngeal cancer: An epidemiological study.

To analyse the risk factors and healing factors of pharyngocutaneous fistula (PCF) in patients with laryngeal cancer after total laryngectomy, and to explore the relevant epidemiology. A retrospective analysis was conducted on laryngeal cancer patients who underwent total laryngectomy in our hospital from January 2010 to December 2022. The 349 patients included in the study were divided into a PCF group of 79 and a non-PCF group of 270. Perform one-way analysis of variance and multivariate logistic analysis on various data of patients included in the statistics, and analyse the risk factors and healing factors of PCF. Smoking, history of radiation therapy for laryngeal cancer, history of chemotherapy for laryngeal cancer, tumour location (larynx, pharynx, oesophagus), preoperative albumin, postoperative proteinaemia, <99 haemoglobin, postoperative haemoglobin, postoperative C-reactive protein (CRP) level are the risk factors for PCF. Also, radiation therapy and postoperative proteinaemia were the main reasons for preventing PCF healing. Smoking history, laryngeal cancer, radiation therapy, albumin, haemoglobin and CRP are risk factors for postoperative PCF after total laryngectomy, while radiation therapy and postoperative hypoalbuminaemia are key factors affecting PCF healing.

The Treatment Effect of Non-Surgical Ear Molding Correction in Children with Mild Cryptotia Deformity.

OBJECTIVE: To investigate the treatment effect of non-surgical ear molding correction in children with mild cryptotia deformity. METHODS: 51 cases were collected from 2016 to 2021. They were divided into four groups (6 months-1 year group, 1-3 years group, 3-6 years group, and ≥6 years group). The effective rate, recurrence rate, complication rate, and treatment duration of non-surgical ear molding correction were analyzed among the four groups. RESULTS: 3 months after the end of corrective treatment, the overall effective rate was 92.2% (47/51), the overall recurrence rate was 7.8% (4/51), and there was statistical significance among the four groups (p = 0.001). The overall complication rate was 2.0% (1/51), and there was no statistical significance among the four groups (p = 1.000). There was statistical significance in the treatment duration among the four groups (p < 0.001), and the mean duration of treatment was positively correlated with the age at treatment (p < 0.001, R = 0.614). CONCLUSIONS: We first propose and recommend that the treatment time window for non-surgical ear molding correction be maximally extended to 6 years old in children with mild cryptotia deformity. There is a high success rate of non-surgical ear molding correction in children with mild cryptotia deformity. The complication rate is low. There is a positive correlation between the mean treatment duration and the age at treatment, and the treatment duration increases with the growth of months. LEVEL OF EVIDENCE: 4 Laryngoscope, 133:2122-2128, 2023.

[Clinical application of orofacial myofunctional therapy combined with muscle functionnal appliance in postoperative rehabilitation of children with OSA].

Objective:To explore the clinical significance of orofacial myofunctional therapy combined with muscle functional appliance in postoperative rehabilitation of children with OSA. Methods:Sixty children were diagnosed as moderate-to-severe OSA with AHI≥5 and underwent adenoid and/or tonsillar surgery. Children were divided into two groups based on whether they were willing to receive orofacial myofunctional therapy and muscle functional appliance after surgery. Lateral cephalogram and portable polysomnography were performed, and the pediatric OSA-18 scale was filled under the guidance of medical staff. The treatment group received combined treatment with orofacial myofunctional therapy and muscle functional appliance. Results:①General condition and subjective symptoms: The total score of OSA-18 in the treatment group was 65.15±11.25 preoperatively and 49.83±7.09 1-month postoperatively, while the score in the control group was 64.69±10.23 preoperatively and 48.07±6.87 1-month postoperatively. The results showed that postoperative sleep, physical symptoms, emotional status, daytime lethargy and energy status of patients, and their influence on their guardians were significantly improved in both groups（P<0.01）. The improvement of sleep disturbance, physical condition, daytime lethargy, the influence on their guardians were greater in the treatment group than in the control group 6-month and 12-month post-operatively（P<0.05）. These findings suggested that oral and facial muscle functional training combined with muscle functional appliance can provide greater improvement in general condition and subjective symptoms in the treatment group. ②PSG: Postoperative AHI and OAI were significantly decreased in both groups, while LSaO2 was significantly increased （P<0.01）, indicating that sleep respiratory function was significantly improved in both groups after treatment. Patients in treatment group showed greater AHI reduction and LSaO2 improvement 6-month and 12-month postoperatively（P<0.01）, indicating that oral and facial muscle functional training combined with muscle functional appliance can provide greater improvement in airway obstruction symptoms and sleep respiration. ③Radiological changes: SNB Angle was increased（P<0.05） and ANB Angle was decreased significantly（P<0.05）, while SPP-SPPW, U-MPW and TB-TPPW increased significantly in airway measurement 6-month and 12-month postoperatively （P<0.01）, indicating that after combined treatment with oral muscle functional training and muscle functional appliance, the mandible was moved forward and rotated clockwise. Conclusion:The combined treatment with oral muscle functional training and muscle functional appliance is more effective in improving oral breathing, upper airway sagittal structure and sleep breathing, and can correct oral habits of children. The long-term effect needs further investigation.

Long noncoding RNA Gm44593 attenuates oxidative stress from age-related hearing loss by regulating miR-29b/WNK1.

Long noncoding RNA has been reported to play important role in various disease. However, the function of lncRNA in age-related hearing loss still unclear. The aim of our study is to investigate the function and mechanism of lncRNA Gm44593 in AHL. ATP content, JC-1 assay, mitochondrial content, cell death rates and dual-luciferase reporter assay were performed to assess the function of lncRNA Gm44593 in HEI-OC1 cells. The expression of lncRNA Gm44593 was significantly upregulated upon H2O2 and starvation treatment. Overexpression of lncRNA Gm44593 manifestly reduced the cell death rates. The ATP content, mtDNA content and mitochondrial membrane potential were alleviated upon overexpression of lncRNA Gm44593. We also proved that miR-29b is the direct target of lncRNA Gm44593. Overexpression of miR-29b completely restored the effect induced by lncRNA Gm44593. In addition, we provided evidences that WNK1 is the direct target of miR-29b. Our research uncovers a potential role of lncRNA Gm44593 in age-related hearing loss. We provide new insights into potential therapeutic targets for the amelioration of age-related hearing loss.

Long non-coding RNA CCAT2 drives the growth of laryngeal squamous cell carcinoma via regulating YAP activity.

Emerging evidence suggests that long non-coding RNA (lncRNA) is closely associated with numerous human diseases, including cancer. However, the functional relevance of lncRNA in human laryngeal squamous cell carcinoma (LSCC) is largely unknown. In the current study, we described CCAT2, a previously unappreciated oncogenic lncRNA in LSCC. CCAT2 was significantly upregulated in human LSCC tissue and serum samples, associated with larger tumor volume, higher clinical stage, and poorer differentiation status. Lentivirus-mediated CCAT2 knockdown notably repressed the cell viability, colony formation, and DNA synthesis rate of LSCC. Screening of transcription factors revealed that YAP/TEAD activity was affected by CCAT2 in LSCC cells. Further, CCAT2 directly binds to YAP protein and blocks the phosphorylation of YAP induced by LATS1, resulting in the nuclear translocation of YAP and the activation of YAP oncogenic targets, such as CTGF, CYR61 and AMOTL2. Importantly, we also confirmed the regulation of CCAT2 on YAP activity in vivo based on nude mice model. Altogether, we identified a novel lncRNA that controls YAP nucleocytoplasmic shuttling and promotes LSCC cell proliferation. Given the importance of YAP in tumorigenesis and progression, our results provide insights to intervene LSCC by targeting the CCAT2/YAP axis.

circRNA circ-CCND1 promotes the proliferation of laryngeal squamous cell carcinoma through elevating CCND1 expression via interacting with HuR and miR-646.

Cyclin D1 (CCND1) is a well-known proliferation promoter that accelerates G1/S transition in cancer. However, the underlying mechanism by which CCND1 is regulated is still largely unknown. In this study, we identified a novel circular RNA (circRNA) derived from CCND1 (circ-CCND1, hsa_circ_0023303) as a key regulator for CCND1. circ-CCND1 was found to be markedly up-regulated in laryngeal squamous cell carcinoma (LSCC) and closely associated with aggressive clinical features and adverse prognosis. Depletion of circ-CCND1 significantly inhibited LSCC cell proliferation in vitro and retarded tumour growth in vivo. Regarding the mechanism, circ-CCND1 physically bound to human antigen R (HuR) protein to enhance CCND1 mRNA stability; on the other hand, circ-CCND1 could act as an effective sponge for miR-646 to alleviate the repression of miR-646 on CCND1 mRNA. As a result, circ-CCND1 post-transcriptionally elevated CCND1 expression via coordinated avoidance of CCND1 mRNA decay, thereby promoting LSCC tumorigenesis. Taken together, our findings uncover the essential proliferation-promoting role of circ-CCND1 through regulation of the stability of CCND1 mRNA in LSCC. Targeting circ-CCND1 may be a promising treatment for LSCC patients.

Long noncoding RNA SOX2-OT facilitates laryngeal squamous cell carcinoma development by epigenetically inhibiting PTEN via methyltransferase EZH2.

Long noncoding RNAs (lncRNAs) play important roles in the initiation and progression of various cancers, including laryngeal squamous cell carcinoma (LSCC). Recently, lncRNA Sox2 overlapping transcript (SOX2-OT) has been identified as an oncogene in various cancers. However, the functional role and the regulatory mechanism of SOX2-OT in LSCC remains unclear. In this study, we found that SOX2-OT expression was increased and negatively correlated with PTEN expression in LSCC tissues. Furthermore, SOX2-OT overexpression promoted LSCC cell proliferation, migration, invasion, and suppressed cell apoptosis in vitro, as well as facilitated the in vivo tumorigenicity. By contrast, SOX2-OT silencing exerted the opposite effect. Mechanically, SOX2-OT interacted with EZH2 and recruited EZH2 to induce H3K27me3 and epigenetically inhibited PTEN expression in LSCC cells. Additionally, EZH2 silencing and PTEN overexpression significantly abrogated the SOX2-OT overexpression-mediated promotion of LSCC cell malignant behavior. Collectively, our findings demonstrate that SOX2-OT inhibits PTEN expression to facilitate LSCC development through EZH2-mediated H3K27me3. © 2019 IUBMB Life, 71(9):1230-1239, 2019.

miR-181a and miR-203 inhibit migration and invasion of laryngeal carcinoma cells by interacting with ATF2.

MicroRNAs (miRNAs) have been recognized to modulate the progression of tumorigenesis by serving as oncogenes or tumor suppressors. Despite the involvement of miR-181a and miR-203 in several cancers as has been substantiated, their roles in laryngeal carcinoma (LC) remain unclear. In this study, the abundances of miR-181a, miR-203 and activating transcription factor 2 (ATF2) mRNA in LC cell lines were detected by RT-qPCR. Western blot was performed to detect the protein levels of N-cadherin, E-cadherin and ATF2. Cell migration and invasion ability were assessed by Trans-well assay. The putative binding sites between miR-181a or miR-203 and ATF2 were predicted using Bioinformatics software and further validated by Dual-Luciferase reporter and RNA immunoprecipitation (RIP) assays. Results showed reduced abundances of miR-181a and miR-203 in LC cell lines. Introduction of miR-181a or miR-203 reduced cell migration and invasion, which was further confirmed by the reduction of N-cadherin and increase of E-cadherin in LC cells. ATF2 was identified to be a potential target of miR-181a and miR-203. Absence of ATF2 overturned the stimulatory effects of anti-miR-181a and anti-miR-203 on cell migration and invasion in LC cells. Our findings suggested that miR-181a and miR-203 attenuated cell migration and invasion ability by directly targeting ATF2 in LC, providing novel insight into the regulatory mechanisms of miR-181a and miR-203 in LC.

Overexpression of Beclin1 inhibits proliferation and promotes apoptosis of human laryngeal squamous carcinoma cell Hep-2.

OBJECTIVE: Beclin1 was previously found to be downregulated in human laryngeal cancer (LC) tissues, and it results in poor prognosis. This study aimed to further confirm the antitumor effects of Beclin1 in LC cell line Hep-2. MATERIALS AND METHODS: Beclin 1 was overexpressed in Hep-2 cells using liposomal transfection and confirmed using reverse transcription polymerase chain reaction and Western blotting. Then, cell proliferation and apoptosis were determined in control (untransfected), empty vector transfected, and Beclin1 overexpressed groups using MTT and flow cytometry procedure, respectively. RESULTS: The expression of the Beclin1 gene in Hep-2 cells was significantly increased after vector transfection compared with control (1.173±0.046 vs 0.453±0.016, P<0.01) and empty vector (1.173±0.046 vs 0.440±0.021, P<0.01). Overexpression of Beclin1 inhibited proliferation at 4 days (0.619±0.051 vs 0.891±0.081 and 0.619±0.051 vs 0.878±0.105, P<0.01), 5 days (0.684±0.078 vs 1.127±0.094 and 0.684±0.078 vs 1.162±0.117, P<0.01), and 6 days (0.725±0.069 vs 1.168±0.103 and 0.725±0.069 vs 1.194±0.097, P<0.01) and promoted apoptosis (14.48%±1.42% vs 4.07%±0.66% and 14.48%±1.42% vs 4.39%±0.80%, P<0.01) in Hep-2 cells in comparison with the control and empty vector groups, respectively. CONCLUSION: Beclin1 may be an underlying target for the treatment of LC. This study has provided some experimental basis for the gene therapy of LC.

miR-200a-3p promotes the proliferation of human esophageal cancer cells by post-transcriptionally regulating cytoplasmic collapsin response mediator protein-1.

The dysregulation of cytoplasmic collapsin response mediator protein 1 (CRMP1) has been reported in lung cancer, medulloblastoma and esophageal cancer. However, the role of CRMP1 and its regulatory mechanisms in esophageal cancer remain unclear. In this study, we demonstrated that CRMP1 expression was downregulated in esophageal cancer tissues and that there were differences in its expression levels in different esophageal cancer cell lines. We found that CRMP1 overexpression inhibited the proliferation of esophageal cancer cells, whereas the silencing of CRMP1 promoted cell proliferation. We performed an analysis of potential microRNA (miRNA or miR) target sites using a commonly used prediction algorithm (TargetScan). The algorithm predicted that miR­200a-3p targets the 3' untranslated region (3'UTR) of CRMP1. Further experiments confirmed this prediction. In addition, we found that miR­200a-3p promoted the proliferation of esophageal cancer cells. Thus, our findings indicate that miR­200a-3p promotes the proliferation of human esophageal cancer cells by post-transcriptionally regulating CRMP1.