CH25H Promotes Autophagy and Regulates the Malignant Progression of Laryngeal Squamous Cell Carcinoma Through the PI3K-AKT Pathway.

BACKGROUND: Laryngeal squamous cell carcinoma (LSCC) is a type of cancer of the respiratory tract that often presents with subtle symptoms at the early stage and is susceptible to recurrence and metastasis. MATERIALS AND METHODS: To find out key regulatory genes involved in LSCC development, we downloaded LSCC-related sequencing datasets for bioinformatics analysis. WGCNA was performed on GSE142083 and differential analysis was conducted on GSE51985 and TCGA-HNSC. Intersection genes were taken from the above three datasets. To confirm the function of genes, we overexpressed and knocked down genes in cells and treated them with autophagy agonist Rapamycin and PI3K-AKT pathway inhibitor. At the cellular level, the expression of CH25H, autophagy-related proteins (LC3 I, LC3 II, p62, and Beclin 1), and PI3K-AKT pathway-related proteins (PI3K, AKT, and p-AKT) were assessed via Western blot; the mRNA level of CH25H was evaluated through qRT-PCR; the cell activity was examined by CCK8; the apoptosis was assessed through flow cytometry; and the cell migration and invasion were assessed through wound healing and Transwell assays. RESULTS: Through bioinformatics analysis, we screened 7 genes (CH25H, NELL2, STC2, TMEM158, ZIC2, HOXD11, and HOXD10). Ultimately, CH25H was selected for follow-up experiments. By detecting CH25H expression in human immortalized keratinocytes (HaCaT) and LSCC cells (Tu-686, SNU899, and AMC-HN-8), it was found out that CH25H expression was higher in HaCaT cells than in LSCC cells. To elucidate the role of CH25H in LSCC development, we overexpressed CH25H in Tu-686 cells and downregulated its expression in AMC-HN-8 cells. CH25H was revealed to reduce the proliferation, activity, invasion, and migration of LSCC cells while increasing their apoptosis levels. Significant changes were also observed in the expressions of autophagy- and PI3K-AKT pathway-related proteins. To further investigate the roles of autophagy and the PI3K-AKT pathway in LSCC development, we respectively employed autophagy agonists and inhibitors targeting the PI3K-AKT pathway to intervene the cells, and found that CH25H regulated the PI3K-AKT pathway to promote autophagy, thus enhancing the apoptosis of LSCC cells. We further investigated CH25H's impact on tumor growth, autophagy, and the PI3K-AKT pathway at the animal level and found that CH25H promoted autophagy of LSCC cells and inhibited the PI3K-AKT pathway, and ultimately inhibiting the progression of LSCC. CONCLUSIONS: In summary, CH25H promotes autophagy and affects the malignant progression of LSCC through the PI3K-AKT pathway.

Study of the significance of the combination of the fibrinogen-albumin ratio and sarcopenia in predicting the prognosis of laryngeal cancer patients undergoing radical surgery.

OBJECTIVE: This study aims to investigate how the impact of preoperative sarcopenia and inflammatory markers for laryngeal cancer patients and develop a new scoring system to predict their prognosis. MATERIALS AND METHODS: Patients who underwent laryngectomy for laryngeal cancer (LC) from December 2015 to December 2020 at the Second Affiliated Hospital of Fujian Medical University were included. Independent prognostic factors were determined using univariate and multivariate analyses. A new scoring system (SFAR) was established based on FAR and preoperative sarcopenia, and statistically analyzed. RESULTS: 198 cases included in this study that met the admission criteria. Multivariate analysis shown that preoperative sarcopenia, pTNM stage, and FAR were independent prognostic factors for laryngeal cancer. Based on these three indicators, we developed the SFAR scoring system. Multivariate analysis showed that SFAR was an independent predictor of laryngeal cancer (p < 0.001). SFAR was then incorporated into a prognostic model that included T-stage and N-stage, and a column-line graph was generated to accurately predict its survival. CONCLUSION: Systemic inflammation and sarcopenia are significantly associated with postoperative prognosis in laryngeal cancer. A new scoring system (SFAR) had implications for improving the prognosis of patients undergoing surgery for laryngeal cancer.

Assessment of quality of voice after 3D conformal hypofractionated radical radiotherapy in early-stage glottic cancer: A prospective study.

PURPOSE: The purpose of this study was to evaluate the short-term outcomes in terms of oncological outcomes, toxicities, and impact of quality of voice on livability of early glottic cancer (EGC) (T1) patients after hypofractionated radiotherapy. MATERIAL AND METHODS: 30 patients of early glottic carcinoma, staged cT1a-T1bN0M0 with Eastern Cooperative Oncology Performance Status (ECOG PS ≤ 2). Exclusion criteria included ECOG PS > 2, T2-T4 disease, and any other head and neck malignancy. Voice Handicap Index-10 (VHI-10) was used to score voice outcomes of patient's pre- and post-radiation. RESULTS: All the subjects were predominantly smokers (80%) having hoarseness of voice as the most common presenting complaint (76.7%) followed by dry cough (13.3%) and foreign body sensation (10%). On posttreatment response assessment, there has been a significant reduction in VHI-10 scores from pre-radiotherapy (RT) to 6, 12, and 24 weeks post-radiation completion in all three domains (functional, emotional, and physical), as well as total scores, with a significant P value (<0.05) being observed. CONCLUSION: The use of modest hypofractionated regimen (63 Gy in 28 fractions) in patients of early glottic disease (T1a and T1b) posttreatment completion provides good oncological outcomes with better quality of voice being observed.

Immunomodulatory role of tumor microenvironment on oncological outcomes in advanced laryngeal cancer.

BACKGROUND: The study evaluated the prognostic impact of the immune microenvironment in LSCC with markers of major immune cells to identify the key determinants of short-term disease-free survival (ST DFS) and reveal factors related to disease progression. METHODS: The study cohort included 61 patients who underwent total laryngectomy, 83.6% of whom were male with a mean age of 64.3 years at the time of surgery. Twenty-five patients had long term DFS (over 5 years), 8 - had moderate DFS (between 2 and 5 years), and 28 had short-term DFS (less than 2 years). Immunohistochemical staining and evaluation were performed on samples collected after the laryngectomy. RESULTS: The samples' assessment revealed that the mean expression of all analysed markers was the highest both in stroma and the tumor compartment for short term DFS (ST DFS) patients. Analysis confirmed that a high stromal density of CD8 cells (p = 0.038) significantly correlated with DFS, and that the increased presence of CD57 cells (p = 0.021) was significantly associated with ST DFS. Moreover, the high density of CD68 cells in the tumor epithelial compartment had a negative prognostic impact on DFS (p = 0.032). Analysis of overall survival in the studied cohort with Kaplan-Meyer curves revealed that a high stromal density of CD68 cells was a significant negative predictor of OS (p = 0.008). CONCLUSIONS: The observed associations of CD68 cells infiltration with progression and prognosis in patients with LSCC provide potential screening and therapeutic opportunities for patients with unfavourable outcomes.

Infantile Subglottic Hemangioma: A Case ReportInfantile Subglottic Hemangioma: A Case Report.

ABSTRACT: Subglottic hemangiomas are rare benign vascular tumors of infancy which involve the airway. It is a subtype of infantile hemangiomas and is a potentially life-threatening condition with a mortality rate of 50% if left untreated. Hence, early intervention in this condition is essential. Here we present a case of a 4-month-old infant, a male infant with a history of cough and noisy breathing requiring multiple hospital visits before eventually being diagnosed with subglottic hemangioma. Due to its similar presentation with other more common respiratory illnesses, the diagnosis can be missed. Oral propranolol is the first-line therapy, which was used successfully in our case.

[A retrospective study on the prognosis of endoscopic surgery for 385 early glottic cancer patients].

Objective: To investigate the prognosis and influencing factors of endoscopic surgery for early glottic carcinoma. Methods: In this retrospective study, we applied the Cox proportional hazards regression model and the random survival forest model to analyze the clinical characteristics of 385 patients [362 males, 23 females, age ranging from 33 to 91 years (62.0±9.6)] who visited the Sixth Medical Center of the General Hospital of the People's Liberation Army from January 2009, to December 2022 and diagnosed with early glottic carcinoma, encompassing variables such as age, gender, T stage, surgical approach, pathological typing, etc. The primary evaluation indicators were overall survival(OS) and disease-free survival rates (DFS). The follow-up duration ranged from 30 to 5,557 days (with a median follow-up time of 1,596 days). Results: After a three-year follow-up, the OS rate for the 385 patients was 95.83%, while the DSF rate was 82.98%. The Cox proportional hazards regression analysis revealed age (HR=2.35, 95%CI: 1.75 to 3.15, P<0.001) and T staging (HR=1.59, 95%CI: 1.13 to 2.23, P=0.019) as predominant factors affecting the OS and DFS. The random survival forest model identified poor tumor differentiation, and high expression of P53 and Ki-67 as predictors of inferior prognosis. Conclusion: Endoscopic surgery for early glottic carcinoma yields favorable short-term OS and reduces short-term recurrence rates, with T-stage emerging as a pivotal factor influencing recurrence. Tumors with poor differentiation and elevated expression of P53 may be indicative of an increased risk of recurrence.

[Analysis of long-term efficacy of CO2 laser partial excision of vocal folds for 599 cases in the treatment of vocal cord leukoplakia].

Objective: To analyze long-term prognosis and influencing factors of recurrence in vocal fold leukoplakia treated by endoscopic cordectomy with CO2 laser. Methods: A retrospective review was conducted on 599 patients with vocal fold leukoplakia [566 males and 33 females, aged 17-84 years (median age 55 years)], undergoing endoscopic cordectomy by CO2 laser under general anesthesia at the Department of Otorhinolaryngology Head and Neck Surgery of Beijing Tongren Hospital, Capital Medical University, from January 2000 to December 2023. The study analyzed the clinical features, pathological grade, surgical methods, postoperative voice function, recurrence, malignant changes and analyzed the long-term efficacy and and its influencing factors. The patients were followed up for a duration of 6-249 months postoperatively, with a median follow-up time of 48 months. SPSS 20.0 software was used for statistical analysis. Results: Among the 599 patients, the histopathological grades included simple squamous cell hyperplasia in (n=264, 44.08%), mild dysplasia (n=96, 16.03%), moderate dysplasia (n=74, 12.35%), severe dysplasia(n=43, 7.81%), carcinoma in situ(n=35, 5.84%), and carcinoma in situ with microinvasion(n=87, 14.52%). The 3-year and 5-year overall recurrence rates were 12.91% and 16.00%, respectively. In patients with precancerous lesion, 3.91% of recurrences presented with an upgraded pathologic grade and 2.34% evolved into carcinoma. The risk of recurrence was higher in lesions involving the anterior commissure, larger lesions, significant reduction/absence of mucosal waves, neovascularization/suspected neovascularization, and pathology of dysplasia/carcinoma in situ/carcinoma in situ with microinvasion(F=44.76,21.54,11.55,8.78,23.20,respectively,P<0.05). Additionally, patients with recurrent disease exhibited higher reflux symptom index scores compared to those without recurrence. Exophytic lesion characteristics and the inability to cease smoking postoperatively were identified as independent risk factors for recurrence with an upgraded pathological grade, with odds ratios of 8.675 and 11.380 times greater than those with non-exophytic lesions and successful smoking cessation, respectively. At the 6-months postoperative assessment, patients who underwent subepithelial cordectomy (typeⅠ) demonstrated a statistically significant increase in fundamental frequency (t=-3.38, P<0.05), and while other voice acoustic parameters were not significantly different.Conversely, those who underwent transmuscular cordectomy (TypeⅢ) exhibited significant alterations in multiple voice acoustic parameters when compared to preoperative values (P<0.05). Furthermore, postoperative vocal fold adhesions developed in 84 patients with 13 of these individuals requiring surgical intervention for adhesion release. Conclusions: Lesions involving the anterior commissure, larger lesions, pathology of dysplasia/carcinoma in situ/carcinoma in situ with microinfiltration, significant reduction/absence of vocal fold mucosal waves, and neovascularization visible under NBI are indicative of an increased risk of recurrence, whereas exophytic lesions and the inability to cease smoking postoperatively significantly increase the risk of recurrence with elevated pathologic grade. Recovery of postoperative voice quality is procedure-dependent, underscoring the importance of surgical approach selection in the management of vocal fold leukoplakia.

Amphiregulin promotes activated regulatory T cell-suppressive function via the AREG/EGFR pathway in laryngeal squamous cell carcinoma.

BACKGROUND: Activated regulatory T cells (aTregs) play a vital role in promoting a tumor immunosuppressive microenvironment in laryngeal squamous cell carcinoma (LSCC). However, the regulatory factors that induce the generation of aTregs are not clear. Herein, we investigated the effect of amphiregulin (AREG) on the production of aTregs in the tumor microenvironment of LSCC. METHODS: Immunohistochemical (IHC) analysis was conducted to examine the expression of AREG and FOXP3, and their association with clinical parameters and patient outcomes was demonstrated. The expression level of EGFRs in three functional subsets of Tregs was assessed, and the induction of CD4+ T cells into aTregs in the presence or absence of AREG or Gefitinib was analyzed using flow cytometry. RESULTS: Our results showed a higher expression level of AREG was significantly related to advanced clinical stage and worse survival, particularly with increased infiltration of Tregs in LSCC tumor tissue. The in vitro study showed that AREG significantly promoted the differentiation of aTregs, and enhanced the inhibitory effect of Tregs on T cell proliferation, which could be reversed by epidermal growth factor receptor (EGFR) inhibitors. In addition, we found that EGFR was highly expressed in aTregs, but not in other subsets of Tregs. It is suggested that AREG might induce aTregs, and enhance the immunosuppressive function of Tregs via the AREG/EGFR signal pathway. CONCLUSIONS: Collectively, this study revealed the role and mechanism of AREG in negative immune regulation, and targeting AREG might be a novel immunotherapy for LSCC.

Investigation of photothermal treatment with 532 nm laser light on laryngeal cancer in an in vivo tumor model.

Laryngeal cancer is the second most common cancer in the upper aerodigestive tract, with its incidence increasing across all ages. The conventional treatments for laryngeal cancer include surgical procedure, radiation, and chemotherapy; however, these treatments can lead to various complications. Photothermal therapy (PTT) using laser light has been employed form cancer effective treatment because of its minimal invasion and short operation time. The current study aims to investigate the feasibility of 532 nm PTT on laryngeal cancer in an invivo tumor model. Ex vivo dosimetry evaluation was conducted to determine the laser irradiation conditions, and HEP-2 tumor bearing mice were used to demonstrate in vivo photothermal effects. In addition, histology and western blot analysis were conducted to verify tumor necrosis and any changes in cancer-associated factors in the tumor tissues. The current in vivo results showed that PTT at 5 W for 40 s and 20 W for 10 s had comparable effects in terms of temperature increase and tumor removal. The 532 nm PTT significantly decreased the remaining tumor and downregulated the expression levels of MMP- 9 and ERK. The current study demonstrated that the 532 nm PTT could be a feasible option for treatment of laryngeal tumor with high power delivery for a short exposure time. Further investigations will confirm the endoscopic application of the 532 nm PTT for the treatment of intralaryngeal tissue prior to clinical translation.

ENAH transcriptionally activated by YY1 promotes growth and invasion of laryngocarcinoma cells through PI3K/AKT signaling.

BACKGROUND: Laryngocarcinoma is a common malignancy in the upper respiratory tract. Enabled homolog (ENAH) is an actin-binding protein that is associated with the development of various cancers. However, its role and mechanism in laryngocarcinoma remain unknown. METHODS: The ENAH level in laryngocarcinoma was examined in silico, in vitro and in vivo. The prognostic analysis of the ENAH level was assessed on laryngocarcinoma patients. Gain- and loss-of-function assays were conducted in AMC-HN-8 and TU686 cells. Sh-ENAH-containing AMC-HN-8 cells were implanted into naked mice. The role and mechanism of ENAH in laryngocarcinoma were investigated by CCK-8, transwell, immunofluorescence, dual luciferase, RT-qPCR, immunohistochemistry, and western blotting experiments. RESULTS: The ENAH level was upregulated in laryngocarcinoma, which predicted a poor prognosis in laryngocarcinoma patients. Gain- and loss-of-function results showed that ENAH promoted proliferation, invasion and EMT of laryngocarcinoma cells. Moreover, ENAH was transcriptionally activated by YY1, and YY1/ENAH axis enhanced these malignant progresses of laryngocarcinoma cells. Besides, ENAH activated the PI3K/AKT pathway, and 740Y-P abolished the accelerative role of ENAH in proliferation, invasion and EMT of laryngocarcinoma cells. Furthermore, knockdown of ENAH reduced tumor size and weight, and the expression level of vimentin and PI3K/AKT pathway in tumor-bearing mice. CONCLUSION: ENAH transcriptionally activated by YY1 promotes cell growth, invasion and EMT of laryngocarcinoma through the activation of PI3K/AKT signaling.

Investigating the Link between STAT4 Genetic Variants, STAT4 Protein Concentrations, and Laryngeal Squamous Cell Carcinoma: A Comprehensive Analysis of Clinical Manifestations.

According to recent research, inflammatory STAT4 and its protein impact may be important factors in developing cancerous diseases. Still unanalyzed is this effect in patients with laryngeal squamous cell carcinoma (LSCC). In the present study, we evaluated four single nucleotide variants (SNVs) of STAT4 (rs10181656, rs7574865, rs7601754, and rs10168266) and STAT4 serum levels to determine their link between LSCC development and its clinical manifestations. A total of 632 men (324 LSCC patients and 338 healthy individuals) were involved in this study. The genotyping was carried out using real-time PCR. Additionally, we measured 80 study subjects' (40 LSCC patients and 40 control subjects) STAT4 protein concentrations using an enzyme-linked immunosorbent assay (ELISA). In our study, the T allele of STAT4 rs7574865 significantly increases the likelihood of LSCC occurrence by 1.4-fold. Additionally, this SNV is associated with higher odds of early-stage disease, T1 size LSCC development, absence of metastasis to neck lymph nodes, and well-differentiated carcinoma. The G allele of rs10181656 is significantly associated with various clinical characteristics of LSCC, increasing the odds of early- and advanced-stage disease by 2.8-fold and 1.9-fold, respectively. Additionally, this allele is linked to an increased likelihood of developing tumors of different sizes and non-metastasized LSCC, as well as poorly differentiated carcinoma, highlighting its potential impact on the development and features of LSCC. Conclusion: The analysis of the STAT4 rs7574865 SNV revealed that the G allele is linked to a more favorable prognosis in LSCC. Additionally, it is hypothesized that the G allele of rs10181656 may be associated with the occurrence of LSCC but may not serve as a sensitive prognostic biomarker for distinguishing between disease stages, cell differentiation, or tumor size.

Laryngeal Preservation Strategies.

Over the last 2 decades, the paradigm of laryngeal cancer management has pivoted toward preserving laryngeal function without sacrificing oncologic outcomes. Transoral laser microsurgery has diminished the role of open laryngeal surgery. For early-stage laryngeal cancer, the common primary modalities are endoscopic laryngeal surgery and narrow field radiation. Total laryngectomy followed by either radiation or chemoradiation is option for advanced laryngeal cancer. In experienced hands and following meticulous patient selection, supracricoid laryngectomy may serve as a viable alternative to total laryngectomy to preserve laryngeal function. Total laryngectomy is still the recommended treatment in those with airway compromise and/or laryngeal dysfunction.

Cancer of the Larynx-20-Year Comparative Survival and Mortality Analysis by Age, Sex, Race, Stage, Grade, Cohort Entry Time-Period, Disease Duration and ICD-O-3 Topographic Primary Sites-Codes C32.0-9: A Systematic Review of 43,103 Cases for Diagnosis Years 1975-2017: (NCI SEER*Stat 8.3.9).

BACKGROUND: .-Laryngeal malignancy, "voice box" cancer, is uncommon with 12,620 estimated new cases and 3770 deaths in the United States in 2021,1 and represents only 6.2% of all respiratory system malignancies. The most significant risk factors are alcohol and tobacco consumption. Almost all cases (98%) of laryngeal cancer arise in the squamous epithelium, and in this analysis more than 75% are of well-or-moderately differentiated histopathology (Grades I&II). Local stage cancer (SEER Historic Staging) was more common than regional and distant stages combined (55.3% vs 44.7%). Tumors may arise above, below or at the level of the vocal folds and are described as supraglottic (encompassing the epiglottis, false vocal cords, ventricles, aryepiglottic fold and arytenoids), the glottis (encompassing the true vocal cords and the anterior and posterior commissures), and the subglottic region. In the National Cancer Institute's Surveillance, Epidemiology, End-Results (NCI-SEER) Data Research, 9 Registries, Nov 2019 Sub (1975-2017),2 laryngeal cancer occurred more commonly in men than in women, 80.7% vs 19.3%, respectively with a 4.2 to 1 ratio. Additionally, there are racial disparities with African Americans presenting at a younger age and having a higher incidence and mortality than Caucasians. In the 1975-2017 period, overall median patient age was 64.4 years with White Americans-64.8 years and Black Americans-61.5 years. Unfortunately, the 5-year relative survival rate has declined 4%, and excess death rate has risen 13% since 1975 with overall incidence declining.As a consequence, observed median survival is approximately 6.5-years for the total study-period pinpointing the need for further specialty research. This study follows the World Health Organization International Classification of Diseases for Oncology-3rd Edition (ICD-O-3)3 topographical identification, coding, labeling and listing of 43,103 patient-cases accessible for analysis in the United States National Cancer Institute's Surveillance, Epidemiology and End Results program (NCI SEER Research Data, 9 Registries, 1975-2017). These are located in 6 primary anatomical sites: C32.0-Glottis, C32.1-Supraglottis, C32.2-Subglottis, C32.3-Laryngeal cartilage, C32.8-Overlapping lesion of larynx, C32.9-Larynx, NOS. OBJECTIVES: .-To update short- and long-term mortality and survival indices, and identify changing risk patterns for laryngeal cancer patients in a retrospective US population-based analysis, 1975-2017, using prognostic data stratified by ICD-O-3 Primary Site, age, sex, race, stage, histologic grade, two cohort entry time-periods (1975-1996 to 1997-2017), and disease duration to 20-years. METHODS: .-SEER*Stat v8.3.94 software (built March 12, 2021) was used to access SEER Research Data, 9 Registries, Nov. 2019 submission (1975-2017). For displaying risk, general methods and standard double decrement life table methodologies for converting and displaying ICD-O-3 coded laryngeal cancer primary site annual data to aggregate average annual mortality and survival units in durational-intervals of 0-1, 1-2, 2-5, 0-5, 5-10, 10-15, and 15-20 years were employed. The reader is referred to the "Registrar Staging Assistant (SEER*RSA)" for local-regional-distant Extent of Disease (EOD) sources used in the development of staging descriptions, and Summary Stage 2018 Coding Manual v2.0 released September 1, 2020. Cancer staging & grading procedural explanations, statistical significance and 95% confidence levels5 are described in previous Journal of Insurance Medicine articles6,7 and other publications.8,9 Poisson confidence intervals at the 95% level based on the number of observed deaths are used in this study but not displayed here to conserve space on the mortality tables. Excluded were all death certificate only and those alive with no survival time. RESULTS: .-Total SEER annual age-adjusted incidence rates from 1980 to 2017 have diminished from 5.25 patient-cases/100,000/year to 2.59/100,000 per year, and in the same period annual age-adjusted US death rates declined from 1.61 deaths/100.000/year to 0.91 deaths/100,000/year (Ref. 10, CSR Tables 12.5-6), However, in the 0-5-year disease durational interval for all staged cases in both cohort time-periods (Table 5), excess death rates (EDR) rose from 80 per 1000 persons per year in the 1975-96 cohort, to 89 per 1000 persons per year in the 1997-17 cohort, (a 10% rise in excess mortality in 42 years). Further, in the 5-10-year disease durational interval, EDR rose from 39 per 1000 persons per year to 45 per 1000 persons per year with corresponding cohort declines in cumulative survival ratios (SR), and overall declines in median observed and relative survival times in the later cohort (not shown). The epidemiologic burden of malignancy is >4-fold higher in males and increases in parallel with aging, peaking after 65 years. The most significant risk factors for laryngeal cancer are tobacco and alcohol consumption. CONCLUSION: .-Although annual incidence and mortality rates from 1980 to 2017 have diminished, there is no concomitant improvement in larynx cancer survival (SR) and mortality (EDR) indices, with rising mortality and diminishing survival in all staged cases at 5-years disease duration between the 1975-96 and 1997-2017 analytic cohorts. Larynx cancer remains a burdensome clinical, social, and public health challenge.

Survival, Surgical, and functional outcomes of transoral laser microsurgery for cT1-T3 supraglottic laryngeal Cancers: A systematic review.

BACKGROUND: This review aimed to investigate the surgical, functional, and oncological outcomes of transoral laser microsurgery supraglottic laryngectomy (TOLM-SGL) for cT1-T3 laryngeal cancers. METHODS: PubMed, Scopus, and Cochrane Library were searched by two independent investigators for studies investigating the surgical, functional, and oncological outcomes of TOLM-SGL using the PRISMA statements. A bias analysis was carried out with MINORS. RESULTS: Twenty-four studies were included (937 patients), including 206 (25.9 %) cT1, 467 (58.7 %) cT2, and 123 (15.4 %) cT3 cases. Most patients were cN0 (63.9 %). The mean hospital stay of TOLM was 10.1 days. Aspiration (5.5 %), and bleeding (5.3 %) were the most prevalent complications. The laryngeal preservation rate was 93.7 %. Temporary tracheotomy was performed in 18.0 % of patients, with a mean time of decannulation of 6.8 days. A feeding tube was placed in 59.9 % of patients. The oral diet restarted after 6.4 days. Definitive gastrostomy was necessary in 2.4 % of cases. The 5-year OS and DFS were 70.1 % and 82.0 %, respectively. Distant metastasis, local, and regional recurrence occurred in 4.6 %, 11.6 %, and 5.1 % of patients. There was an important heterogeneity between studies for inclusion criteria, patient profiles, TOLM indications, and details of surgical, functional, and oncological outcomes. CONCLUSION: TOLM supraglottic laryngectomy is a safe, and effective procedure associated with adequate functional, surgical, and oncological outcomes. Future studies are needed to define the place of TOLM in advanced LSCC; the role and timing of concomitant bilateral neck dissection, the indications of tracheotomy and feeding tube.

Polyvinyl Alcohol Capped Silver Nanostructures for Fortified Apoptotic Potential Against Human Laryngeal Carcinoma Cells Hep-2 Using Extremely-Low Frequency Electromagnetic Field.

Purpose: : Polyvinyl alcohol-capped silver nanostructures (cAgNSs) were investigated in order to enhance the cytotoxicity, pro-apoptotic, and oxidant patterns of in human laryngeal carcinoma Hep-2 cells by employing a 50 mT electromagnetic field (LEMF) for 30 min. Methods: Wet chemical reduction was used to synthesize the cAgNSs, and after they had been capped with polyvinyl alcohol, they were specifically examined for particle size analysis and structural morphology. To visualize how the silver may attach to the protein targets, a molecular docking study was conducted. Estimation of cytotoxicity, cell cycle progression supported by mRNA expression of three apoptotic-promoting genes and one apoptotic-resisting. Results: Particle size analysis results were a mean particle size of 157.3±0.5 nm, zeta potential value of -29.6 mV±1.5 mV, and polydispersity index of 0.31±0.05. Significantly reduction of IC50 against Hep-2 cells by around 6-fold was concluded. Also, we obtained suppression of the proliferation of Hep-2 cells, especially in the G0/G1 and S phases. Significant enhanced mRNA expression revealed enhanced induced CASP3, p53, and Beclin-1 mediated pro-apoptosis and induced NF-κB mediated autophagy in Hep-2 cells. Augmented levels of GR, ROS and MDA as oxidative stress biomarkers were also obtained. HE staining of Hep-2 cells exposed to cAgNSs and LEMF confirmed the enhanced apoptotic potential comparatively. Conclusion: By conclusion, the developed nano-sized structures with the aid of extremely-low frequency electromagnetic field were successful to fortify the anti-cancer profile of cAgNSs in Hep-2 cells.

Non-canonical RNA-binding protein ANXA11 regulates microRNA resorting into small extracellular vesicles to mediate cisplatin resistance.

The sensitivity of laryngeal squamous cell carcinoma (LSCC) to chemotherapy shows large heterogeneity. The role of miRNA in small extracellular vesicles (sEV) in chemotherapy resistance is under investigation. However, the regulation and sorting mechanism of sEV miRNAs remains unclear. In this study, small RNA sequencing was used to explore miRNA expression profiles in sEV of LSCC after cisplatin stimulation; RNA pull-down, mass spectrometry, and EMSA were used to clarify the binding of candidate RNA-binding protein (RBP) and candidate miRNA. Immunostaining and microRNA fluorescence in situ hybridization were performed to identify how candidate RBP affects miRNA stability and nuclear/cytoplasmic distribution. In vivo experiments were performed to verify the biological functions and response to cisplatin of candidate RBP. We found that cisplatin stimulation induced increased expression of miR-148a-3p and sEV sorting. ANXA11 binds to miR-148a-3p in a sequence-specific manner. ANXA11 inhibits tumor cell proliferation and drug resistance by binding to and retaining miR-148a-3p. Cisplatin stimulation reduced ANXA11 expression and promoted miR-148a-3p efflux through sEV pathways. ANXA11 overexpression reduced in vivo tumor proliferation and cisplatin-resistance. Taken together, ANXA11 mediates cisplatin resistance through sEV miRNA resorting. Mechanically, ANXA11 binds to miR-148a-3p in a sequence-specific manner to regulate its resorting and thus influences tumor proliferation and chemoresistance.

Spindle cell lipoma, a rare entity affecting the larynx: A case report.

Spindle cell lipoma is a very rare occurrence in the larynx and can be cured by complete excision. In this case report we present the case of a 71-year-old female who presented to the otorhinolaryngology outpatient department of Northwest General Hospital and Research Centre, Peshawar, with complaints of occasional irritation and a foreign body sensation in the throat while swallowing, for the last three years. She had undergone a surgical procedure 30 years back for the same complaint and remained asymptomatic till three years back. On examination, through fibre-optic laryngoscope, the attending surgeon saw an abnormal mass arising from the aryepiglottic folds of the larynx. An excisional biopsy was performed through micro-laryngoscopy. The patient's symptoms subsequently improved and she is currently doing well. Histopathological reports confirmed it as spindle cell lipoma.