Exploration of molecular mechanism of Selaginella moelledorffii Hieron. in treatment of laryngeal cancer based on network-based pharmacology, molecular docking techniques and experimental validation / 中国药理学通报

Aim To explore the molecular mechanism of Selaginella moelledorffii Hieron. in the treatment of laryngeal cancer. Methods According to the relevant literature reports, the chemical constituents of S. moellendorffii were obtained, and the active ingredients were screened out through the SwissADME database, and the targets were screened through the PharmMapper database. The laryngeal cancer-related targets were collected by searching OMIM and other databases, and the Venny 2.1.0 online platform was used to obtain the intersection of the two. Protein interaction analysis of the potential targets was performed using the STRNG platform. GO functional analysis and KEGG pathway analysis was carried out using DAVID database. Visual networks were built with Cytoscape 3.8.0 software. Molecular docking was validated by SYBYL-X 2. 0 software. MTT method, Hoechst 33258 staining method and Western blotting were also used for validation. Results At the molecular level, a total of 110 active ingredients of S. moellendorffii and 82 drug targets were screened out, 1,608 targets related to laryngeal cancer, and intersection of 34 targets. GO analysis yielded 135 entries, and KEGG analysis yielded a total of 61 pathways. Molecular docking results showed that the 11 key active ingredients such as 2", 3"-dihydrooch-naflavone wood flavonoids and 4 core target proteins such as MAPK1 had 95. 5% of good docking activity. At the cellular level, SM-BFRE was screened for its strongest inhibitory effect on laryngeal cancer cell proliferation through MTT assay. Furthermore, Hoechst 33258 staining showed that the decrease in Hep-2 cell viability produced by SM-BFRE was related to cell apoptosis. Finally, Western blot verified that SM-BFRE inhibited PI3K/Akt/NF through inhibition- K B/COX-2 pathway to induce apoptosis in laryngeal cancer cells. Conclusions To sum up, it fully reflects the multicomponent, multi-target, and multi-channel synergistic effect of S. moellendorffii in the treatment of laryngeal cancer, and provides a theoretical reference for further elucidation of the mechanism of action of S. moellendorffii in the treatment of laryngeal cancer.

Prognostic and predictive role of soluble programmed death ligand-1 in head and neck cancer

Abstract Objectives The aim of the study was to investigate clinical significance of soluble PD-L1 (sPD-L1) serum level in head and neck cancer and to evaluate its role as a possible prognostic and predictive biomarker. Methods A prospective analysis of sPD-L1 levels in 60 patients diagnosed and treated due to malignant and non-malignant lesions in the region of head and neck was performed in peripheral blood by an ELISA test. Results The range of sPD-L1 in the study group was 0.16-1.63 ng/mL, mean 0.64 ± 0.32. There were no differences in the mean sPD-L1 regarding patients' age, sex, and the localization of the lesion. Statistically significant difference was revealed in the average sPD-L1 level (p = 0.006) depending on the histopathological advancement of the lesions, 0.704 ± 0.349 and 0.512 ± 0.177 respectively in the malignant and benign group. The separate analysis of laryngeal lesions confirmed statistical difference in sPD-L1 (p = 0.002) for the malignant lesions (0.741 ± 0.353) compared with the benign (0.489 ± 0.175). The sPD-L1 level of 0.765 ng/mL or higher, revealed 35% sensitivity and 95.5% specificity for the diagnosis of head and neck malignant lesions (AUC = 0.664, 95% CI 0.529‒0.8, p-value = 0.039). The 1-year DFS was 83.3% in the group of patients with low sPD-L1 levels (< 0.765 ng/mL) and 53.8% in patients with high sPD-L1 (≥0.765 ng/mL). The 2-year OS were 68% and 69.2% respectively in both groups. The log-rank test confirmed statistically significant prognostic value of sPD-L1 level for 1-year DFS (p-value = 0.035). Conclusions sPD-L1 is a promising prognostic and early recurrence predictive biomarker for head and neck cancers, most significantly for laryngeal lesions. Level of evidence 3.

Comparative Analysis of the Quality of Life in the Pretreatment of Head and Neck Cancer Patients According to Tumor Site

Abstract Introduction Head and neck cancer (HNC) and its treatment can cause physical, psychological, and quality of life (QoL) damage, because it can disturb the physiology of eating, breathing, speaking, and compromise self-image. Objective To evaluate the QoL of the pretreatment of patients diagnosed with head and neck cancer according to the anatomical location of the tumor. Methods A descriptive, cross-sectional study was performed on a sample of 144 patients undergoing pretreatment for cancer from February 2017 to July 2019. The University of Washington QoL Questionnaire (version 4) was used to assess the QoL. The anatomical location data were obtained from medical records. The ANOVA test was used to compare the differences in QoL according to tumor location. Results A total of 144 participants were included, 66 (45.5%) of whom had the primary tumor located in the mouth. The median age of the patients was 62 years, with a higher prevalence of male (75.7%), Black (78.5%), single/divorced/widowed people (59%), and illiterates (32.6%); most of them were smokers (84.7%) and alcohol drinkers (79.2%). The mean QoL score was 830 for mouth cancer, 858 for pharynx cancer, and 891 for laryngeal cancer patients. Conclusion Based on the results of this study, it can be concluded that the QoL of patients with head and neck cancer was not influenced by tumor location. The most affected domains in the three groups were pain, appearance, chewing, swallowing, and speech (p < 0.05).

A CLINICAL STUDY OF PREVALENCE OF LARYNGEAL CANCER IN ELDERLY AND ITS ASSOCIATION WITH SMOKING AND ALCOHOL

Laryngeal Cancer refers to cancerous tumor that develops in Larynx, for example, glottis, supraglottic region and subglottis. The purpose of this present study was to investigate the prevalence of Laryngeal Cancer in elderly patients attending a tertiary care centre. A detailed survey was performed to analyze whether Laryngeal Cancer is associated with smoking or alcohol. The study revealed that Laryngeal Cancer is more common in elderly individuals who smoke and drink alcohol. Avoidance of alcohol intake and smoking will be a milestone to reduce the incidence of Laryngeal Cancers and associated mortality.

Identification of biomarkers in laryngeal cancer by weighted gene co-expression network analysis / 中南大学学报(医学版)

OBJECTIVES@#Laryngeal cancer (LC) is a globally prevalent and highly lethal tumor. Despite extensive efforts, the underlying mechanisms of LC remain inadequately understood. This study aims to conduct an innovative bioinformatic analysis to identify hub genes that could potentially serve as biomarkers or therapeutic targets in LC.@*METHODS@#We acquired a dataset consisting of 117 LC patient samples, 16 746 LC gene RNA sequencing data points, and 9 clinical features from the Cancer Genome Atlas (TCGA) database in the United States. We employed weighted gene co-expression network analysis (WGCNA) to construct multiple co-expression gene modules. Subsequently, we assessed the correlations between these co-expression modules and clinical features to validate their associations. We also explored the interplay between modules to identify pivotal genes within disease pathways. Finally, we used the Kaplan-Meier plotter to validate the correlation between enriched genes and LC prognosis.@*RESULTS@#WGCNA analysis led to the creation of a total of 16 co-expression gene modules related to LC. Four of these modules (designated as the yellow, magenta, black, and brown modules) exhibited significant correlations with 3 clinical features: The age of initial pathological diagnosis, cancer status, and pathological N stage. Specifically, the yellow and magenta gene modules displayed negative correlations with the age of pathological diagnosis (r=-0.23, P<0.05; r=-0.33, P<0.05), while the black and brown gene modules demonstrated negative associations with cancer status (r=-0.39, P<0.05; r=-0.50, P<0.05). The brown gene module displayed a positive correlation with pathological N stage. Gene Ontology (GO) enrichment analysis identified 77 items, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis identified 30 related signaling pathways, including the calcium signaling pathway, cytokine-cytokine receptor interaction, neuro active ligand-receptor interaction, and regulation of lipolysis in adipocytes, etc. Consequently, central genes within these modules that were significantly linked to the overall survival rate of LC patients were identified. Central genes included CHRNB4, FOXL2, KCNG1, LOC440173, ADAMTS15, BMP2, FAP, and KIAA1644.@*CONCLUSIONS@#This study, utilizing WGCNA and subsequent validation, pinpointed 8 genes with potential as gene biomarkers for LC. These findings offer valuable references for the clinical diagnosis, prognosis, and treatment of LC.

Baicalein promotes laryngeal cancer cell death and inhibits invasion via miR-125b-5p/IRF4 axis / 中国临床药理学与治疗学

AIM: To investigate the mechanism of baicalin-induced apoptosis in human laryngeal cancer cells. METHODS: AMC-HN-8 cells were selected for the study, and baicalin was applied to the cells at different concentrations (0, 10, 30, 100, and 300 μmol/L), and the half-inhibitory concentration (IC50) was measured by the CCK-8 method. Bax, cleaved-caspase-3, Cyto-c, IRF4 protein expression by protein blotting (Western blot); miR-125b-5p and IRF4 expression by RT-qPCR. Dual-luciferase reporter gene validation of Targetscan prediction (binding of miR-125b-5p to IRF4-3'UTR); apoptosis and necrosis inhibitors explore the way baicalein induces death in laryngeal cancer cells. AMC-HN-8 was then divided into blank group, baicalein (IC50), miR-125b-5p inhibitor group, baicalein + inhibitor NC group, baicalein+miR-125b-5p inhibitor group, and cell invasion and clone formation assays to detect cell invasion and proliferation ability, respectively. Apoptosis was detected by flow cytometry. RESULTS: Baicalein inhibited the proliferation of AMC-HN-8 cells in a dose-dependent manner with an IC50 value of 47.31 μmol/L. Compared with the blank group, 47.31 μmol/L baicalin induced apoptosis and inhibited cell invasion, while upregulating the expression of miR-125b-5p and suppressing the mRNA and protein levels of IRF4. The luciferase results showed that the miR-125b-5p mimic was able to inhibit the activity of the IRF4-3'UTR promoter relative to the NC mimic (mimic) group. Baicalein induces laryngeal cancer cell death in an apoptotic manner. In addition, the combination of 47.31 μmol/L baicalin and miR-125b-5p inhibitor affected the behavior of AMC-HN-8 cells, showing that compared with the blank group, the baicalin group showed a decrease in the number of cell clones, weakened invasion ability, and increased apoptosis; the miR - 125b-5p inhibitor group showed an increase in the number of cell clones, enhanced invasion ability and decreased apoptosis. The baicalin+ inhibitor NC group was consistent with baicalin, with no significant effect of inhibitor NC on cell behavior. The cloning, invasion, and apoptosis of cells in the baicalin+miR-125b-5p inhibitor group were intermediate between the baicalin and miR-125b-5p inhibitor groups. CONCLUSION: Baicalin inhibits the proliferation of AMC-HN-8 cells, and the mechanism may be related to miR-125b-5p targeting to inhibit the expression of IRF4, inducing the pro-apoptotic proteins Bax, cleaved-caspase3, and Cyto-c, and inhibiting the apoptosis suppressor protein Bcl-2 thereby inducing apoptosis.

Relato de caso: actinomicose mimetizando neoplasia laríngea / Case report: actinomycosis mimicking laryngeal neoplasia / Caso clínico: actinomicosis que simula una neoplasia laríngea

A actinomicose é uma infecção rara causada por bactérias do gênero Actinomyces sp. que são importantes constituintes das flora comensal de animais e humanos, colonizando a orofaringe, o trato respiratório superior, o trato gastrointestinal e o trato genital feminino. Diante de uma lesão na mucosa associada a um desequilíbrio nas defesas do organismos, as bactérias se infiltram nas estruturas, gerando uma infecção. As áreas mais comumente afetadas por esta patologia correspondem às regiões cervicofacial (50%), abdominal (20%) e torácica (15 a 20%), sendo o envolvimento laríngeo bastante raro (< 5% dos casos), com poucos casos descritos na literatura. Este estudo visa relatar o caso de um paciente de 76 anos com actinomicose mimetizando neoplasia de laringe, através da revisão do prontuário, descrevendo o quadro clínico e evidenciando os exames de imagem realizados na investigação com o objetivo de alertar os profissionais de saúde e compartilhar informações sobre esta patologia para que possa fazer parte do diagnóstico diferencial de doenças granulomatosas e infecciosas da laringe, principalmente em pacientes com mais de 50 anos, visando o diagnóstico precoce e o tratamento adequado.

Actinomycosis is a rare infection caused by bacteria of the genus Actinomyces sp. which are important constituents of the commensal flora of animals and humans, colonizing the oropharynx, the upper respiratory tract, the gastrointestinal tract and the female genital tract. Faced with a lesion in the mucosa associated with an imbalance in the organism's defenses, the bacteria infiltrate the structures, generating an infection. The areas most commonly affected by this pathology correspond to the cervicofacial (50%), abdominal (20%) and thoracic (15 to 20%) regions, and laryngeal involvement is quite rare (< 5% of cases), with few cases described in the literature. This study aims to report the case of a 76-year-old patient with actinomycosis mimicking laryngeal neoplasm, through the medical record review, describing the clinical picture and evidencing the imaging tests performed in the investigation with the purpose of alerting health professionals and sharing information about this pathology so that it can be part of the differential diagnosis of granulomatous and infectious diseases of the larynx, especially in patients older than 50 years, aiming at early diagnosis and appropriate treatment.

La actinomicosis es una infección rara causada por bacterias del género Actinomyces sp. que son componentes importantes de la flora comensal de animales y humanos, colonizando la orofaringe, el tracto respiratorio superior, el tracto gastrointestinal y el tracto genital femenino. Ante una lesión de la mucosa asociada a un desequilibrio de las defensas del organismo, las bacterias se infiltran en las estructuras, generando una infección. Las áreas más comúnmente afectadas por esta patología corresponden a las regiones cervicofacial (50%), abdominal (20%) y torácica (15 a 20%), siendo la afectación laríngea bastante rara (< 5% de los casos), con pocos casos descritos en la literatura. El presente estudio tiene como objetivo relatar el caso de una paciente de 76 años con actinomicosis simulando neoplasia laríngea, a través de la revisión de la historia clínica, describiendo el cuadro clínico y mostrando los estudios de imagen realizados en la investigación, con el propósito de alertar a los profesionales de la salud y compartir información sobre esta patología para que forme parte del diagnóstico diferencial de las enfermedades granulomatosas e infecciosas de la laringe, particularmente en pacientes con edad superior a 50 años, con vistas al diagnóstico precoz y tratamiento adecuado.

Asociación entre la infección por virus del papiloma humano y el carcinoma de laringe en adultos de 35-75 años de edad en el instituto nacional del cáncer rosa emilia sánchez pérez de tavarez (incart), en el período agosto 2012 - enero 2019, santo domingo, república dominicana / Association between human papillomavirus infection and laryngeal carcinoma in adults between 35 and 75 years of age at the Rosa Emilia Sánchez Pérez de Tavarez National Cancer Institute (INCART), in the period August 2012 - January 2019, Santo Domingo, Dominican Republic

Introducción: el carcinoma de cabeza y cuello comprende un grupo de neoplasias que comparten un origen anatómico similar. Ocupan el sexto lugar mundial entre las neoplasias de todas las localizaciones, y el sitio afectado con mayor frecuencia es la cavidad oral. Ahora se reconoce el rol del virus del papiloma humano como factor independiente en el desarrollo de estas neoplasias. En los últimos 15 años se ha observado un incremento en la incidencia de carcinoma de células escamosas inducido por virus del papiloma humano (VPH). Materiales y métodos: este es un estudio observacional de tipo descriptivo transversal. Se realizó un análisis documental de los expedientes clínicos de los pacientes que cumplan con los criterios de inclusión. Resultados: en este estudio fue encontrada una prevalencia del 14 % del VPH en cánceres laríngeos. El tipo 16 del VPH fue el único identificado y descrito entre los expedientes clínicos de este estudio y, este último, con una prevalencia de un 20 %. Discusión: el VPH puede estar involucrado en el desarrollo de algunos cánceres de laringe y su rol puede ser más predominante en hombres mayores de 50 años.

Introduction: Head and neck carcinoma comprises a group of neoplasms with similar anatomical origins. They occupy the sixth place in the world among the neoplasias of all the locations, and the most affected site is the oral cavity. The role of the Human Papillomavirus as an independent factor in the development of these neoplasms is now recognized. In the last 15 years, an increase in the incidence of squamous cell carcinoma induced by Human Papillomavirus (HPV) has been observed. Methods: This is an observational study of transversal descriptive type. A documental analysis of the clinical files of the patients that meet the inclusion criteria was performed. Results: in this study, a 14 % prevalence of HPV in laryngeal cancers was found. Type 16 HPV was the only one identified and described in the clinical files of this study and the latter with a prevalence of 20 % Discussion: HPV may be involved in the development of some laryngeal cancers and its role may be more predominant in men older than 50 years.

Particularités du cancer du larynx chez le sujet jeune / Epidemiological and clinical predictive factors of success in septoplasty: Study of 250 patients

Discuter à travers une étude descriptive ainsi qu'une revue de la littérature, les particularités cliniques, démographiques et pronostiques des patients de moins de 45 ans, ayant un cancer du larynx. Materiels et Methodes: Il s'agit d'une étude rétrospective descriptive portant sur des patients atteints d'un cancer du larynx, âgés de moins de 45 ans suivis dans le service d'ORL et de chirurgie cervico-faciale du CHU Habib Bourguiba Sfax durant la période s'étendant de 1989 à 2018. Resultats: Nous avons trouvé 31 patients avec une prédominance masculine. Un cancer dans la famille a été trouvé dans 16,12% des cas sans corrélation statistique avec le stade avancé de la maladie. Une importante intoxication tabagique a été trouvée (96%). Trois patients avaient une laryngite chronique et un patient une papillomatose laryngée avec des lésions de dysplasie. Les motifs de consultation étaient dominés par la dysphonie (87%). La maladie a été classée en stades avancés dans 70% des cas. Le traitement chirurgical était préconisé chez 87% des patients et la préservation fonctionnelle chez 38,7%. Le taux de survie globale et sans maladie étaient respectivement, à un an de 96% et 84%, à 3 ans de 87% et 76%, et à 5 ans de 77% et 75% Conclusion: Notre travail n'a pas permis de retenir de différence en termes de données cliniques, de l'évolution de la maladie, de l'algorithme thérapeutique ni du pronostic entre les jeunes patients et les plus âgés

Treatment Strategies and Prognostic Factors in Patients with Stage T3 and T4 Laryngeal Carcinoma / 肿瘤防治研究

Objective To investigate the selection of treatment strategies and prognostic factors for patients with stage T3 and T4 laryngeal carcinoma. Methods A total of 132 patients with stage T3 and T4 laryngeal cancer admitted to our hospital from March 2010 to March 2019 were retrospectively selected as research objects. According to the different treatment strategies, the patients were divided into simple surgery group (group A, 57 cases), simple chemoradiotherapy group (group B, 32 cases), and surgery combined with chemoradiotherapy group (group C, 43 cases). The general data and clinicopathological features of the three groups were compared, and a survival curve was drawn by the Kaplan–Meier method. The 3-year survival rates of the three groups were compared. Then, the same 132 patients were divided into survival and death groups. The clinical data of the two groups were compared, and the prognostic factors were analyzed by multivariate logistic regression. A back propagation (BP) neural network model was constructed, and its differentiation and accuracy were evaluated. Results The proportions and 3 year survival rates of patients with poor differentiation, lymphatic vascular invasion, and involvement of lymph nodes outside the capsule in group C were significantly higher than those in groups A and B (P < 0.05). The 3 year survival rate of 132 patients was 68.94%(41/132). Poor differentiation, N2-N3 stage, lymphatic vascular invasion, and involvement of lymph nodes outside the capsule were risk factors for death (P < 0.05), whereas surgery combined with radiotherapy and chemotherapy were protective factors (P < 0.05). The BP neural network model exhibited good discrimination and high accuracy. Conclusion Surgery combined with radiotherapy and chemotherapy can significantly improve survival rate in patients with poor differentiation, lymphatic vascular invasion, and involvement of lymph nodes outside the capsule. Close attention should be paid to patients with stage N2-N3 in the formulation of reasonable treatment strategies.

Impact of resection margin status and revision transoral laser microsurgery in early glottic cancer: analysis of organ preservation and local disease control on a cohort of 153 patients / Impacto do status da margem de ressecção e microcirurgia transoral a laser de revisão no câncer glótico inicial: análise da preservação do órgão e controle local da doença em uma coorte de 153 pacientes

Abstract Introduction Transoral laser microsurgery represents the treatment of choice for early glottic cancer. Its use and effectiveness are mainly related to laryngeal exposure and deep extension of tumor. Histopathologic assessment of surgical margin presents a main issue about transoral laser microsurgery and complete oncological excision. Objective The aim was to analyze the impact of revision surgery on organ preservation and local disease control in patients with early glottic cancer treated by transoral laser microsurgery. Methods We carried out a retrospective study on a cohort of 153 patients with early glottic cancer (Tis, T1, T2) treated by transoral laser microsurgery. Resection margins were classified as follows: "free" if macroscopic margin-tumor distance was at least 2 mm, as "close" if it was less than 2 mm and "positive" if the margin was involved by carcinoma. Patients were divided into two groups: patients with free resection margins (Group A) and patients with positive, close or not-evaluable resection margins (Group B). Group A (36) underwent periodic followup. Group B (117) underwent a second look laser CO2 2 months after surgery. Fifteen patients of Group A with suspected persistence of carcinoma during followup underwent a second laser resection after a time interval of 4-8 months after first surgery. Overall survival, disease-free survival, disease-specific survival, ultimate local control with laser alone and organ preservation rates were estimated. Results Five-year overall survival rate and 5-year disease-specific survival were 100% in both groups. The five-year laryngeal preservation rate was 100% in Group A and 95.2% in Group B. Five-year disease-free survival was 92.15% and 5-year ultimate local control with laser alone in 92.15% of patients. Conclusion This study has demonstrated that revision Transoral Laser Microsurgery is able to confirm the oncological radicality in most cases, even in the case of positive, close or non-evaluable margins. Considering our results, according to our experience, the second look with CO2 laser is a therapeutic strategy to consider, even in the case of close or non-evaluable as well as positive margins.

Resumo Introdução A microcirurgia transoral a laser representa o tratamento de escolha para o câncer glótico inicial. Seu uso e sua eficácia estão relacionados principalmente à exposição laríngea e à profundidade da extensão do tumor. A avaliação histopatológica da margem cirúrgica representa uma das principais questões sobre a microcirurgia a laser transoral e excisão oncológica completa. Objetivo Analisar o impacto da cirurgia de revisão na preservação de órgãos e controle local da doença em pacientes com câncer glótico inicial tratados por microcirurgia a laser transoral. Método Fizemos um estudo retrospectivo em uma coorte de 153 pacientes com câncer glótico inicial (Tis, T1, T2) tratados por microcirurgia transoral a laser. As margens de ressecção foram classificadas da seguinte forma: livre, se a distância margem macroscópica-tumor fosse de pelo menos 2 mm; próxima, se fosse menor do que 2 mm; e positiva se a margem fosse afetada pelo carcinoma. Assim, os pacientes foram divididos em dois grupos: com margens de ressecção livres (grupo A) e com margens de ressecção positivas, próximas ou não avaliáveis (grupo B). O grupo A (36) foi submetido a seguimento periódico. O grupo B (117) foi submetido a uma cirurgia de revisão com laser de CO2 2 meses após a cirurgia. Quinze pacientes do grupo A com suspeita de persistência do carcinoma durante o seguimento foram submetidos a uma segunda ressecção a laser após um intervalo de 4 a 8 meses após a primeira cirurgia. A sobrevida global, a sobrevida livre de doença, a sobrevida doença-específica, o controle local final com laser isolado e as taxas de preservação de órgãos foram estimados. Resultados A taxa de sobrevida global em cinco anos e a sobrevida doença-específica em 5 anos foi de 100% em ambos os grupos. A taxa de preservação laríngea em cinco anos foi de 100% no grupo A e 95,2% no grupo B. A sobrevida livre de doença em cinco anos foi de 92,15% e o controle local final em 5 anos com laser isolado em 92,15% dos pacientes. Conclusões A revisão da microcirurgia transoral a laser é capaz de confirmar a radicalidade oncológica na maioria dos casos, mesmo em caso de margens positivas, próximas ou não avaliáveis. Considerando nossos resultados, de acordo com a nossa experiência, a cirurgia de revisão (second look) com laser de CO2 é uma estratégia terapêutica a ser pensada mesmo no caso de margens próximas ou não avaliáveis, bem como em margens positivas.

Role and mechanism of miR-548-3p/ DAG1 in the occurrence and malignant transformation of laryngeal carcinoma / Papel y mecanismo de miR-548-3p/DAG1 en la aparición y transformación maligna del carcinoma de laringe

Abstract The AMC-HN-8 cell line and the primary human laryngeal epithelial cell lines were utilized in this work to explore the molecular mechanism of miR-548-3p regulating the gene DAG1 to induce the occurrence and malignant transformation of laryngeal carcinoma. Non-coding RNA miR-548- 3p overexpression plasmid, interference plasmid and blank plasmid were constructed, and the plasmids were transfected into AMC-HN-8 cells, respectively. Meanwhile, a non-transfected plasmid group and a human laryngeal epithelial primary cell group were set up. Five groups of cells were named as NC (Normal control), Model, Ov-miR-548-3p, Sh-miR-548-3p and Blank-plasmid group. The luciferase reporter experiment was used to analyze the regulation characteristics of hsa-miR-548-3p on dystrophin-associated glycoprotein 1 (DAG1). Immunofluorescence was used to analyze the relative expression characteristics of the protein DAG1. The cell cloning experiment was used to analyze the proliferation characteristics of AMC-HN-8. The scratch healing test was used to analyze the migration ability of AMC-HN-8. The transwell test was used to analyze the invasion ability of AMC-HN-8. The RT-PCR was used to analyze the expression level of miR-548-3p. Western blot experiments were used to analyze the expression of protein DAG1, laminin α2 (LAMA2) and utrophin (UTRN). The luciferase report experiment and immunofluorescence test found that the expression of DAG1 and miR-548-3p are positively correlated. Cell cloning, scratching and migration experiments identified that the activity of laryngeal cancer cells was positively correlated with the expression of DAG1. The results of Western blot analysis further strengthened the above conclusions. Through carrying out research on the cellular levels, our work has demonstrated that miR-548-3p regulated the content of protein DAG1, and then further induced malignant transformation of laryngeal carcinoma.

Resumen En este trabajo se utilizaron la línea celular AMC-HN-8 y la línea celular epitelial laríngea humana primaria, para explorar el mecanismo molecular regulador del miR-548-3p sobre el gen DAG1 para inducir la aparición y la transformación maligna del carcinoma laríngeo. Se construyeron el plásmido de sobreexpresión de miR-548-3p de ARN no codificante, el plásmido de interferencia y el plásmido en blanco, y los plásmidos se transfectaron en células AMCHN-8 respectivamente. Mientras tanto, se establecieron un grupo de plásmidos no transfectados y un grupo de células primarias epiteliales laríngeas humanas. Se nombraron cinco grupos de células como NC (control normal), modelo, OvmiR-548-3p, Sh-miR-548-3p y grupo de plásmido en blanco. El experimento indicador de luciferasa se utilizó para analizar las características de regulación de hsa-miR-548-3p en la glicoproteína 1 asociada a distrofina (DAG1). Se utilizó inmunofluorescencia para analizar las características de expresión relativa de la proteína DAG1. El experimento de clonación celular se utilizó para analizar las características de proliferación de AMC-HN-8. Se utilizó la prueba de cicatrización por rascado para analizar la capacidad de migración de AMC-HN-8. La prueba de transwell se utilizó para analizar la capacidad de invasión de AMCHN-8. Se utilizó RT-PCR para analizar el nivel de expresión de miR-548-3p. Se usó un experimento de transferencia Western (Western blot) para analizar las expresiones de la proteína DAG1, laminina α2 (LAMA2) y utrofina (UTRN). El experimento de reporte de luciferasa y la prueba de inmunofluorescencia encontraron que la expresión de DAG1 y miR-548-3p están positivamente correlacionadas. Los experimentos de clonación celular, rascado y migración, identificaron que la actividad de las células cancerosas de laringe se correlacionó positivamente con la expresión de DAG1. Los resultados del análisis de transferencia Western fortalecieron aún más las conclusiones anteriores. A través de la investigación a nivel celular, nuestro proyecto ha demostrado que miR-548-3p regula el contenido de la proteína DAG1 y luego induce la transformación maligna del carcinoma de laringe.

Results of radiotherapy in squamous cell laryngeal cancer: A tomotherapy center experience

Background: Laryngeal cancer is a common type of head and neck cancer (HNC). Radiotherapy (RT) is a mainstay for curative treatment. Intensity-modulated RT (IMRT) is a standard technique today, as it provides of higher survival and local control and lower normal tissue toxicity. One of IMRT devices is helical tomotherapy (HT). The HT treatment results of HNC patients have been reported in few studies. We aimed to investigate the results of squamous cell laryngeal carcinoma patients treated with helical tomotherapy. Methods: Forty-five laryngeal cancer patients were selected according to the inclusion criteria. Radiotherapy (RT) plans were set in the Hi-Art HT planning system. Image-gated RT (IGRT) technique was used. Appropriate patients received simultaneous cisplatin. Treatment response rates were evaluated at the post-RT third and sixth months. Survival times were calculated with the Kaplan–Meier method. The factors affecting the treatment results were evaluated using Log-rank and Cox regression tests. A P value of less than 0.05 was accepted as statistically significant. Results: The median age was 65 (28–84) years. The median symptom duration was 6 (1–60) months. The RT dose for the early and the locally advanced disease was median 63 Gy (60.75–66) and 66 Gy (60–70), respectively. The RT interruption was median two (0-20) days. The patients were followed up to 25 (1–45) months. Grade 2 xerostomia and dysphagia rates were 55% and 7%, respectively. The 3-year estimates of overall survival (OS), disease-free survival (DFS), metastasis-free survival (DMFS), and locoregional recurrence-free survival (RRFS) were 71.7%, 60.4%, 84.9%, and 68.5%, respectively. In univariate analysis, the presence of N2 disease was a negative prognostic for DFS (P = 0.05) and DMFS (P = 0.003). RT interruption >2 days was a negative prognostic for OS (P = 0.005), DFS (P = 0.02), and RRFS (P = 0.023). In the multivariate analysis, symptom duration >6 months was found to be the only significant factor for DFS (P < 0.05). Conclusion: Intensity-modulated radiation with HT achieved comparable clinical outcomes with acceptable toxicity in laryngeal carcinoma.

Survival results of 3786 patients with stage I or II laryngeal squamous cell carcinoma: a study based on a propensity score / Resultados de sobrevivência de 3.786 pacientes com carcinoma epidermoide de laringe em estádio I ou II: um estudo baseado em escore de propensão

Abstract Introduction Laryngeal squamous cell carcinoma is the second most prevalent malignancy among head and neck tumors, and the treatment of patients with stage I or II disease can be performed with surgery or radiation therapy. National population studies describing therapeutic results comparing these modalities are unusual, but they can be very important to direct treatment guidelines. Objective To evaluate the survival results of patients with laryngeal squamous cell carcinoma at clinical stages I or II, according to the main therapeutic modalities used. Methods Cross-sectional, population-based study using the database of Fundação Oncocentro de São Paulo from January 2000 to March 2019. Inclusion criteria were patients with laryngeal squamous cell carcinoma in clinical stages cT1-2N0. To compensate for the non-random allocation of patients and the imbalance between confounding variables between groups, we used the propensity score methodology. Results A total of 3786 patients met the inclusion criteria. Regarding the cT stage, there were 2171 patients (57.3%) with cT1 tumors. Patients in the public health system had a longer time between diagnosis and treatment (p< 0.001). The analysis by propensity score showed that patients treated with surgery had a tendency towards better disease-specific survival (p = 0.012). Comparing radiotherapy alone versus its combination with radiochemotherapy, radiotherapy alone showed a tendency towards a better survival rate (p< 0.001). Conclusion Analysis by propensity score identified better results for disease-specific survival in patients with laryngeal squamous cell carcinoma at clinical stages I and II treated by surgery when compared to radiotherapy.

Resumo Introdução O carcinoma epidermoide de laringe é o segundo mais prevalente entre os tumores de cabeça e pescoço e o tratamento dos pacientes com doença em estádio I ou II pode ser feito com cirurgia ou radioterapia. Estudos populacionais nacionais que descrevem os resultados terapêuticos e compararam essas modalidades são poucos frequentes, mas podem ser muito importantes para orientar diretrizes de tratamento. Objetivo Avaliar os resultados de sobrevida dos pacientes com carcinoma epidermoide de laringe em estádios clínicos I ou II de acordo com as principais modalidades terapêuticas usadas. Método Estudo transversal de base populacional com a base de dados da Fundação Oncocentro de São Paulo de janeiro de 2000 a março de 2019. Os critérios de inclusão foram pacientes com carcinoma epidermoide de laringe em estádios clínicos cT1-2N0. Para compensar pela alocação não randômica dos pacientes e pelo desequilíbrio entre variáveis confundidoras entre os grupos, usamos a metodologia do escore de propensão. Resultados Preencheram os critérios de inclusão 3.786 pacientes. Em relação ao estádio cT, houve 2.171 pacientes (57,3%) com tumores cT1. Os pacientes do sistema único de saúde (SUS) apresentaram um maior tempo entre o diagnóstico e o tratamento (p < 0,001). A análise por escore de propensão mostrou que os pacientes submetidos a cirurgia apresentaram uma tendência de melhor sobrevida doença específica (p = 0,012). Comparando-se radioterapia isolada versus combinação com radioquimioterapia, o tratamento isolado demonstrou tendência a melhor taxa de sobrevida (p< 0,001). Conclusão A análise por escore de propensão identificou melhores resultados de sobrevida doença específica em pacientes com carcinoma epidermoide de laringe estádios clínicos I e II tratados por cirurgia quando comparados a radioterapia.

Pyroptosis-related lncRNAs Predict Prognosis of Laryngeal Cancer / 肿瘤防治研究

Objective To construct a prognostic model of laryngeal cancer based on pyroptosis-related lncRNAs. Methods Transcriptome expression and clinical data of patients with laryngeal cancer were downloaded from TCGA database. Differentially-expressed pyroptosis-related lncRNAs were selected using Wilcox rank sum test and Spearman correlation analysis. LncRNAs associated with patients' prognosis were further selected using univariate Cox analysis (P < 0.05), and a prognostic model was established using multivariate Cox regression. ROC curve was used to assess the sensitivity and specificity of this model. Results There were a total of 483 differentially-expressed pyroptosis-related lncRNAs in laryngeal cancer tissues, compared with normal laryngeal tissues (|logFC|≥1, FDR < 0.05). Univariate Cox analysis showed that 23 differentially-expressed lncRNAs were associated with prognosis. Multivariate Cox regression analysis finally obtained a prognostic model based on 10 lncRNAs for predicting the survival of laryngeal carcinoma patients. AUC showed that the model had a good predictive ability (AUC > 0.8). Conclusion Pyroptosis-related lncRNAs can be used to predict the prognosis of patients with laryngeal cancer.

Comparación de sutura mecánica con sutura manual de la faringe en laringectomía total / Comparison of mechanical suture with manual suture of the pharynx in total laryngectomy

Introducción: La sutura mecánica es una opción para el cierre de la faringorrafia en laringectomía total por cáncer de laringe. Objetivo: Comparar el uso de sutura mecánica lineal con sutura manual de la faringe durante la laringectomía total por cáncer de laringe. Material y Método: Se realizó un estudio de tipo experimental prospectivo. Se compararon dos grupos pacientes: Los pacientes con sutura mecánica desde 2018 a marzo de 2020 y los pacientes con sutura manual previa a enero de 2018 en el Servicio de Otorrinolaringología del Hospital Barros Luco. Se analizaron fístula faringo-cutánea posoperatoria (FFC), tiempo operatorio, estadía hospitalaria y costo. Resultados: El estudio incluyó a 14 pacientes, cada grupo con n = 7. El grupo con sutura mecánica presento 0% de FFC y el grupo sutura manual 28%. El grupo con sutura mecánica reinicio alimentación a los 7 días y el grupo con sutura manual en promedio a los 11,5 (7-23) días. (p = 0,0023). El tiempo promedio de cirugía para el grupo experimental es de 288 ± 37,4 minutos y con sutura manual 311 ± 32,4 minutos. (p = 0,0176). El promedio de hospitalización para el grupo experimental fue de 11 ± 2,6 días (9 a 16), y para el grupo control fue de 21 ± 14 días (10 a 49) (p < 0,0001). Conclusión: La sutura mecánica es un procedimiento fácil de usar y seguro. Existiría un beneficio en el uso de sutura mecánica para el cierre faríngeo al compararlo con la sutura manual al disminuir el número de FFC, el tiempo operatorio y los días de hospitalización.

Introduction: Mechanical suture is an option for the closure of the pharyngorrhaphy in total laryngectomy due to laryngeal cancer. Aim: To compare the use of linear mechanical suture with manual pharyngeal suture during total laryngectomy for laryngeal cancer. Material and Method: A prospective experimental study was carried out. Two patient groups were compared: patients with mechanical suture from 2018 to March 2020 and patients with manual suture prior to January 2018 at the otorhinolaryngology service of the Barros Luco Hospital. Postoperative pharyngocutaneous fistula (FFC), operative time, hospital stay and cost were analyzed. Results: The study included 14 patients, each group with n = 7. The group with mechanical suture presented 0% of FFC and the group with manual suture 28%. The group with mechanical suture restarted feeding at 7 days and the group with manual suture on average at 11.5 (7-23) days (p = 0.0023). The average surgery time for the experimental group was 288 ± 37.4 minutes, while for the group with manual suture was 311 ± 32.4 minutes (p = 0.0176). The mean hospitalization for the experimental group was 11 ± 2.6 days (9 to 16), and for the control group it was 21 ± 14 days (10 to 49) (p < 0.0001). Conclusion: The mechanical suture is an easy to use and safe procedure. There would be a benefit in the use of mechanical suture for pharyngeal closure when buying it with manual suture by reducing the number of FFCs, operative time and days of hospitalization.

Time between Diagnosis and Treatment of Hypopharynx and Larynx Cancer: Are Longer Delays Associated with Higher Discrepancy between Clinical and Pathological Staging?

Abstract Introduction At the time of diagnosis, treatment strategies for cancer are largely based upon clinical staging. However, discrepancy between clinical and pathological staging has been reported. Objective To assess the rate of staging discrepancy in Laryngeal and Hypopharyngeal Squamous Cell Carcinoma (LHSCC), the potential influence of higher interval of time from diagnosis to primary surgical treatment, and whether this has any impact on survival outcomes. Methods Retrospective study of patients with LHSCC proposed for primary surgical treatment. Results The study population included 125 Caucasian patients with LHSCC. The level of agreement between clinical and pathological tumor staging was moderate (Cohen's Kappa: 0.400; p < 0.001) and similar result was found for node staging (Cohen' Kappa: 0.520; p < 0.001). The mean time between diagnosis and surgical treatment was 26.66 days and no statistically significant influence was found with staging discrepancy. The sample presented a 5-year Overall Survival (OS) of 58.2% and a Disease-specific survival (DSS) of 72.6%. No statistically significant impact of staging discrepancy on survival was found. Conclusion For advanced LHSCC, based on the findings of physical examination, endoscopy and imaging, is possible to achieve a moderate accuracy between clinical and pathological staging which allows a reliable counselling and treatment planning. Interval of time under 3-4 weeks between diagnosis and surgical treatment does not influence the rate of discrepancy. However, almost 30% of staging discrepancy is expected due to false negatives of imaging and limitations of physical exams.

Laringectomía total en el adulto mayor, Guantánamo 2015-2019 / Laringectomia total em idosos, Guantánamo 2015-2019 / Total laryngectomy in older adult, Guantanamo 2015-2019

Introducción: El cáncer laríngeo en el adulto mayor tiene una incidencia elevada por múltiples factores que se incrementan con la edad, pero con un tratamiento específico adecuado como la laringectomía total permiten mejorar y elevar la calidad de vida de estos pacientes. Objetivo: Caracterizar el resultado de las intervenciones por laringectomía total realizadas en pacientes adultos mayores en el servicio de Otorrinolaringología del Hospital General Docente "Dr. Agostinho Neto", provincia Guantánamo, en el período de enero de 2015 a diciembre de 2019. Método: Se realizó un estudio descriptivo, retrospectivo de corte transversal en el total de pacientes (n=32) de 60 años y más a los que se les realizó laringectomía total durante el periodo estudiado. Las variables estudiadas fueron: edad, sexo, hábitos tóxicos, condicionantes prequirúrgicas, técnica quirúrgica y complicaciones más frecuentes. Los datos se obtuvieron de las historias clínicas. Resultados: La mayoría de los pacientes correspondió al sexo masculino (90,6 porciento) entre 70-74 años; el 71,8 porciento de los pacientes fumaba. La laringectomía total combinada con tiroidectomía unilateral o bilateral y vaciamiento cervical selectivo fue la técnica quirúrgica más empleada y el faringostoma la complicación predominante. Conclusiones: La caracterización de los resultados de la laringectomía total en el adulto mayor portador de carcinoma epidermoide laríngeo aporta datos que permiten una mejor atención integral a este grupo poblacional(AU).

Introduction: Laryngeal cancer has a high incidence in older adult due to various factors which has been increasing in frequency with age. However, assuming an adecuated and specific treatment like total laryngectomy, allows improving and enhancing the patient's quality of life. Objective: To characterize the outcomes of total laryngectomy in older adult patients treated in the otorhinolaryngology service at the Hospital General Docente "Dr. Agostinho Neto" in Guantánamo, from January 2015 to December 2019. Method: A descriptive, retrospective, and cross-sectional study was conducted on the total of patients (n=32) 60 and older, who underwent total laryngectomy during the mentioned period. Variables used were as follow: age, sex, toxic habits, preoperative conditions, surgical technique and, most frequent complications. Data was gathered from medical records. Results: Male patients 70 to 74 years were predominant (90.6 percent) and most of them smoked (71.8 percent). Total laryngectomy combined with unilateral or bilateral thyroidectomy and selective neck dissection was the most common surgical technique used and pharyngostoma was the predominant complication. Conclusions: The characterization of total laryngectomy outcomes in older adult with laryngeal squamous cell carcinoma provides important data for better comprehensive care in this population group(AU).

Introdução: O câncer de laringe em idosos apresenta alta incidência devido a múltiplos fatores que aumentam com a idade, mas com um tratamento específico adequado como a laringectomia total, melhoram e melhoram a qualidade de vida desses pacientes. Objetivo: Caracterizar o resultado da laringectomia total realizada em pacientes idosos no serviço de Otorrinolaringologia do Hospital General Docente "Dr. Agostinho Neto", província de Guantánamo, no período de janeiro de 2015 a dezembro de 2019. Método: Estudo descritivo, retrospectivo e transversal em todos os pacientes (n=32) com 60 anos ou mais que realizaram laringectomia total. durante o período estudado. As variáveis estudadas foram: idade, sexo, hábitos tóxicos, condições pré-cirúrgicas, técnica cirúrgica e complicações mais frequentes. Os dados foram obtidos dos prontuários médicos. Resultados: A maioria dos pacientes era do sexo masculino (90,6 porcento) entre 70-74 anos; 71,8 porcento dos pacientes fumavam. A laringectomia total combinada com tireoidectomia unilateral ou bilateral e dissecção cervical seletiva foi a técnica cirúrgica mais utilizada e a faringostomia a complicação predominante. Conclusões: A caracterização dos resultados da laringectomia total em idosos com carcinoma espinocelular de laringe fornece dados que permitem uma melhor assistência integral a este grupo populacional(AU).

Which risk factors are associated with stomal recurrence after total laryngectomy for laryngeal cancer? A meta-analysis of the last 30 years / Quais fatores de risco estão associados à recorrência estomal após laringectomia total para câncer de laringe? Uma meta-análise dos últimos 30 anos

Abstract Introduction Stomal recurrence is a troublesome complication after total laryngectomy. Despite a large number of studies having been performed, there is still controversy about which risk factors are most significant for the development of stomal recurrence. Objective The objective of the present meta-analysis was to analyze the potential factors leading to stomal recurrence after total laryngectomy. Methods PubMed, Web of Science, Cochrane Library, and Ovid databases were systematically searched using multiple search terms. Eighteen studies with 6462 patients were identified. The quality of evidence was assessed by The National Institute for Health and Clinical Excellence. Results The results showed that, tumor subsite (supraglottic vs. subglottic, RR = 0.292, 95% CI 0.142-0.600, p = 0.001; glottic vs. subglottic, RR = 0.344, 95% CI 0.175-0.676, p = 0.002), T stage (RR = 0.461, 95% CI 0.286-0.742, p = 0.001), preoperative tracheotomy (RR = 1.959, 95% CI 1.500-2.558, p < 0.001) were the high-risk factors associated with the development of stomal recurrence. Conclusion From the results of our study, tumor subsite, T stage and preoperative tracheotomy were the significant risk factors for stomal recurrence. Methodologically high-quality comparative investigations are needed for further evaluation.

Resumo Introdução A recorrência estomal pós-laringectomia total e é uma complicação de difícil manejo. Apesar de um grande número de estudos ter sido feito, ainda há controvérsias sobre quais fatores de risco são mais significativos para o seu desenvolvimento. Objetivo O objetivo da presente meta-análise foi analisar os fatores potenciais para recorrência estomal após a laringectomia total. Método As bases de dados PubMed, Web of Science, Cochrane Library e Ovid foram sistematicamente pesquisadas com vários termos de busca. Dezoito estudos com 6.462 pacientes foram identificados. A qualidade da evidência foi avaliada pelo National Institute for Health and Clinical Excellence. Resultados Os resultados mostraram que o subsítio tumoral (supraglótico vs. subglótico, RR = 0,292, IC 95% 0,142 ± 0,600, p = 0,001; glótico vs. subglótico, RR = 0,344, IC 95% 0,175 ± 0,676, p = 0,002), estágio T (RR = 0,461, IC 95% 0,286 ± 0,742, p = 0,001), traqueotomia pré-operatória (RR = 1,959, IC 95% 1.500 ± 2.558, p < 0,001) foram os fatores de alto risco associados ao desenvolvimento de recorrência estomal. Conclusão Com base nos resultados de nosso estudo, o subsítio do tumor, o estágio T e a traqueotomia pré-operatória foram fatores de risco significativos para recorrência estomal. São necessárias investigações comparativas metodologicamente de alta qualidade para maior avaliação.

Well-Being of Caregivers of Patients with Laryngeal Cancer Treated by Radiotherapy

Abstract Introduction Laryngeal cancer is the second most common head and neck cancer, but no study to date reports exclusively on the well-being of the caregivers of patients with laryngeal cancer treated by radiotherapy. Objective The present prospective pilot study aims to describe the impact of the work on the well-being of the caregivers of patients with laryngeal cancer. Methods The caregivers of patients undergoing radiotherapy with curative intent for laryngeal cancer were included in the present study, and they were asked to fill out a specific questionnaire, while the patients filled out the Hospital Anxiety and Depression Scale (HADS) following diagnosis but prior to treatment. Results A total of 50 caregivers were included, 20% of which were male, and 80%, female. In total, 62% (n = 31) of the caregivers were spouses, while the remaining were daughters/sons (n = 11; 22%), siblings (n = 6; 12%), or they were classified as "other" (n = 2; 4%). The female caregivers scored lower (worse) on the depression/worry domain, and this was statistically significant (p = 0.047); they also reported a statistically significant higher negative impact on employment (p = 0.011) compared with the male caregivers. In general, the caregivers of patients with late-stage tumors tended to report lower (worse) scores on all domains (except hospital contact) comparedwith the caregivers of patients with early-stage tumors. No other differences were observed regarding the patients' age, gender, tumor site or their HADS score. Conclusion The diagnosis and treatment of laryngeal cancer impacts the caregiver's psychological well-being, particularly that of female caregivers. This should be taken into consideration when the patients begin the hospital treatment pathway. However, larger studies are needed to target resources more appropriately.