Laryngeal Tissue Research Articles

Twenty Eight Days in the Life of a Vocal Pathology: A Case Study of Videolaryngostroboscopy, Acoustic, and Perceptual Variability

The purpose of this investigation was to observe laryngeal tissue and vocal function changes over the course of 28 days in a single participant diagnosed by a laryngologist with bilateral nodules. Laryngeal imaging, acoustic variables and perceptual assessments of voice quality, and perceived vocal effort were obtained every morning for 28 consecutive days. A daily journal of occupational and recreational voice use as well as menstruation and alcohol consumption was maintained each day. It was hypothesized that the laryngeal pathology would appear more extensive and the vocal function measures obtained would be worse following extensive voice use. Laryngeal imaging, acoustic variables, and perceptual measures quantified provided evidence to support the study hypotheses. The size, extent, and asymmetry of the bilateral vocal pathologies observed were more extensive on days following occupational and recreational vocal loading. Acoustic and perceptual measures obtained correlated with the laryngeal tissue changes observed. Study findings support a more holistic approach to laryngeal pathology diagnosis that is based on a more thorough understanding of vocal loading considerations up to 48 hours prior to laryngeal endoscopy to better understand the pathophysiology of the observed lesion(s) for most accurate clinical diagnosis.

LncRNA WTAPP1 promotes proliferation of laryngeal carcinoma cells through regulating microRNA-592.

The aim of this work was to investigate the mechanism by which long non-coding RNA (lncRNA) WTAPP1 promotes the malignant progression of laryngeal cancer. In this study, quantitative real-time polymerase chain reaction (qRT-PCR) examined the expression of lncRNA WTAPP1 in 49 pairs of tumor tissue specimens and paracancerous normal ones collected from laryngeal cancer patients. Subsequently, in the laryngeal squamous cell carcinoma cell lines AMC-HN-8 and Hep-2, WTAPP1 overexpression and knockdown vectors were constructed using lentivirus, and cell counting kit-8 (CCK-8), cell colony formation and 5-ethynyl-2'-deoxyuridine (EdU) assays were carried out to analyze the impact of lncRNA WTAPP1 on the function of laryngeal cancer cells. Finally, Luciferase reporting assay and recovery experiments were carried out to further explore whether lncRNA WTAPP1 has an impact on the malignant progression of laryngeal cancer via modulating microRNA-592. QRT-PCR results revealed a significantly higher expression of lncRNA WTAPP1 in tumor tissues of patients with laryngeal cancer than that in adjacent normal ones. Compared with patients with low expression of WTAPP1, those with higher expression had a more advanced pathological stage. Meanwhile, the proliferation ability of cells in sh-WTAPP1 group was remarkably attenuated when compared with that in sh-NC group. In addition, microRNA-592 and WTAPP1 mRNA levels were found negatively correlated in laryngeal carcinoma tissue specimens. Luciferase reporter gene assay indicated that WTAPP1 can be targeted by microRNA-592 through certain binding sites. Moreover, we demonstrated through some recovery experiments that WTAPP1 can indeed serve as an oncogene accelerating the malignant progression of laryngeal cancer through the modulation of microRNA-592. LncRNA WTAPP was markedly highly expressed both in laryngeal carcinoma tissues and cell lines, which was also found to be closely relevant to the pathological stage of laryngeal cancer patients. Additionally, lncRNA WTAPP1 is able to enhance the proliferation capacity of laryngeal carcinoma cells via regulating microRNA-592.

Kanserli ve Sağlıklı Larenks Dokularında ICP-MS Yöntemi ile Eser Elementler Analizi

Eser elementler vücudun düzgün çalışması için gereklidir. Biyolojik element konsantrasyonundaki değişiklikler çok sayıda patolojik durumda gözlenmiştir. Bu çalışmanın amacı, larenks dokularındaki eser element konsantrasyonunun değerlendirilmesidir. Çalışmada larenks kanseri teşhisi konmuş 9 hastanın kanserli larenks dokuları ve aynı hastaların sağlıklı larekns dokuları kullanılmıştır. Larenks dokularındaki Al, Cr, Mn, Co, Ni, Cu, Zn, As, Se, Ag, Cd, Sb ve Pb seviyeleri, Endüktif Eşleşmiş Plazma Kütle Spektrometresi (ICP-MS) kullanılarak belirlenmiştir. Sonuçlar kontrol grubu ile karşılaştırıldı ve larenks kanserli dokularda Cu seviyesinin sağlıklı dokulara göre anlamlı derecede (p <0.05) yüksek olduğu ve Al, Cr, Mn, As, Se, Ag ve Sb elementlerinin derişiminin ise sağlıklı dokulara göre anlamlı derecede düşük olduğu bulundu. Bu sonuçlar, larenks kanseri etyolojisinde eser elementlerin önemli rolleri olduğunu düşündürmektedir.

Anti-cancer effect of miR-139-3p on laryngeal squamous cell carcinoma by targeting rab5a: In vitro and in vivo studies

BackgroundLaryngeal squamous cell carcinoma is the second most common head and neck squamous cell carcinoma. Nowadays, as traditional treatment methods are gradually limited, the development of new treatment methods needs to be resolved. This study aimed to investigate the role of microRNA(miR)-139-3p in laryngeal squamous cell carcinoma, and further explored the underlying mechanism. MethodsIn this study, we first used quantitative real time polymerase chain reaction (qRT-PCR) to detect the level of miR-139-3p in laryngeal squamous cell carcinoma tissue. Then, TargetScan and dual luciferase reporter assay were used to explore and verify whether rab5a was a direct target of miR-139-3p. Thereafter, the expression of miR-139-3p and rab5a in laryngeal squamous cell carcinoma cell line SNU46 was changed by transfection with miR-139-3p mimic or rab5a-plasmid. Then, SNU46 cell proliferation, cell apoptosis, cell cycle, cell migration and cell invasion were determined using Cell Counting Kit-8 (CCK-8), flow cytometry, scratch assay and Transwell assay, respectively. Finally, mouse tumor formation experiments were used to test whether miR-139-3p still exerted its role in inhibiting laryngeal squamous cell carcinoma in vivo. ResultsCompared with the adjacent normal tissues, miR-139-3p significantly down-regulated in laryngeal squamous cell carcinoma tissue. It was confirmed by dual luciferase reporter experiment that rab5a was a direct target of miR-139-3p. Moreover, the up-regulation of miR-139-3p could effectively inhibit the proliferation, migration and invasion of laryngeal squamous cell carcinoma cells, and induced cell cycle arrest and apoptosis. In the molecular level study, we found that up-regulation of miR-139-3p inhibited the expression of rab5a in SNU46 cells. In addition, the protein and mRNA expression of factors related to cell migration, invasion, proliferation and apoptosis, such as integrin β1, Focal adhesion kinase (FAK), paxillin, B cell lymphoma-2 (bcl-2), nuclear factor-kappaB (NF-κB), vascular endothelial growth factor (VEGF), matrix metalloproteinase 9 (MMP9), in SNU46 cells were changed after miR-139-3p up-regulation. Consistent with the results of in vitro studies, in vivo experiments showed that miR-139-3p mimic inhibited laryngeal squamous cell carcinoma tumor growth. All the effects of miR-139-3p on laryngeal squamous cell carcinoma were reversed by rab5a over-expression. ConclusionmiR-139-3p could inhibit laryngeal squamous cell carcinoma by targeting rab5a both in vitro and in vivo.

The Association Between Endotracheal Tube Size and Aspiration (During Flexible Endoscopic Evaluation of Swallowing) in Acute Respiratory Failure Survivors.

To determine whether a modifiable risk factor, endotracheal tube size, is associated with the diagnosis of postextubation aspiration in survivors of acute respiratory failure. Prospective cohort study. ICUs at four academic tertiary care medical centers. Two hundred ten patients who were at least 18 years old, admitted to an ICU, and mechanically ventilated with an endotracheal tube for longer than 48 hours were enrolled. Within 72 hours of extubation, all patients received a flexible endoscopic evaluation of swallowing examination that entailed administration of ice, thin liquid, thick liquid, puree, and cracker boluses. Patient demographics, treatment variables, and hospital outcomes were abstracted from the patient's medical records. Endotracheal tube size was independently selected by the patient's treating physicians. For each flexible endoscopic evaluation of swallowing examination, laryngeal pathology was evaluated, and for each bolus, a Penetration Aspiration Scale score was assigned. Aspiration (Penetration Aspiration Scale score ≥ 6) was further categorized into nonsilent aspiration (Penetration Aspiration Scale score = 6 or 7) and silent aspiration (Penetration Aspiration Scale score = 8). One third of patients (n = 68) aspirated (Penetration Aspiration Scale score ≥ 6) on at least one bolus, 13.6% (n = 29) exhibited silent aspiration, and 23.8% (n = 50) exhibited nonsilent aspiration. In a multivariable analysis, endotracheal tube size (≤ 7.5 vs ≥ 8.0) was significantly associated with patients exhibiting any aspiration (Penetration Aspiration Scale score ≥ 6) (p = 0.016; odds ratio = 2.17; 95% CI 1.14-4.13) and with risk of developing laryngeal granulation tissue (p = 0.02). Larger endotracheal tube size was associated with increased risk of aspiration and laryngeal granulation tissue. Using smaller endotracheal tubes may reduce the risk of postextubation aspiration.

Abstract 4252: 3D label-free, real-time, intelligent-augmented chemistry-sensitive imaging to identify parathyroid adenoma and hyperplasia and to classify parathyroid glands and recurrent laryngeal nerve during surgery

Abstract Background: Accidental excision or damage to a healthy parathyroid gland (PG) during thyroidectomy and the inability to identify diseased PGs during parathyroidectomy can result in postsurgical complications. Meanwhile, hyperparathyroidism includes primary and secondary hyperparathyroidism, which can cause damage in skeleton, kidney, and heart, with bone pain and renal colic. Therefore, it is critical to distinguish healthy parathyroid when the operation is being processed since the extent of the parathyroidectomy depends on the accuracy of the diagnosis. Methods: Human cancerous and healthy thyroid tissues were obtained from Shanghai General Hospital. Porcine thyroid glands, recurrent laryngeal nerves (RLN), adipose tissues, muscle tissues, lymph nodes, and parathyroid glands were harvested in fresh. Each sample was divided into two sections, one for CARS imaging and the other for hematoxylin and eosin staining (H&E). The H&E images were then classified by a senior pathologist. Some imaging analysis methods were performed to quantify some parameters, such as overall normalized intensity, regional normalized intensity, etc., to quantitatively differentiate the six types of tissues. Deep-learning algorithms were developed for auto-diagnosing thyroid cancer and for differentiating parathyroids, thyroids, lymph nodes, fats, RLN, and muscles. Results: Human thyroid cancer tissue can be identified immediately and automatically through CARS imaging combined with deep-learning algorithms. Parathyroid adenoma/hyperplasia presented as enlarged chief cells and clear cells with reduced fat droplets compared with normal parathyroid. The CARS identification results were confirmed by an experienced pathologist. All six different porcine tissues can be identified by the same system, and the correspondence to human equivalents was confirmed by the pathologist. The accuracy of both identification is above 90%. Quantitative analysis was performed to differentiate parathyroid, thyroid, lymph node, and fat, and differentiate RLN from muscle. Significance: The intelligent-augmented label free imaging system combining CARS imaging and deep-learning algorithm can reliably identify thyroid cancer tissue and differentiate it from thyroid gland, lymph node, and adipose tissue morphologically, without staining, accurately, and immediately. Therefore, this system is a potential tool for quantitatively differentiating parathyroid glands and RLN from surrounding tissues and identifying thyroid cancer tissue in real-time for diagnosis and during parathyroid surgery. Ultimately, the imaging technique can be incorporated into handheld probe for use in clinical diagnosis, such as cancer margin detection during surgery for minimal dissection and reducing nerve impairment during prostate cancer surgery. Citation Format: Jiasong Li, Ye Wang, Yunjie He, Mary C. Farach-Carson, Stephen T.C. Wong. 3D label-free, real-time, intelligent-augmented chemistry-sensitive imaging to identify parathyroid adenoma and hyperplasia and to classify parathyroid glands and recurrent laryngeal nerve during surgery [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4252.

The Down-Regulation of TrkB Alleviates the Malignant Biological Behavior and Cancer Stem-Like Property of Laryngeal Cancer.

BackgroundThis study aimed to evaluate the effect of TrkB down-regulation on the malignant biological behavior and stem-like characteristics of laryngeal cancer.MethodsThe relationship was analyzed between TrkB and clinicopathological parameters in patients with laryngeal cancer. The mRNA expressive levels of TrkB and miR-10a-5p were detected by qRT-PCR in laryngeal cancer tissues and cell lines. In vitro, Hep-2 and AMC-HN-8 cell proliferation, apoptosis and stem-like properties were detected by colony formation assay, flow cytometry, sphere formation, and Western blot, respectively. In vivo, the BALB/c nude mice model was used to evaluate the effect of TrkB on tumor growth.ResultsThe results showed that TrkB was related to smoking history, clinical stage, and lymph node metastasis, but had nothing to do with the gender, age, and tumor location of patients with laryngeal cancer. TrkB was highly expressed and miR-10a-5p was lowly expressed in laryngeal cancer tissues and cell lines. Down-regulation of TrkB inhibited Hep-2 and AMC-HN-8 cell proliferation and sphere formation as well as enhanced apoptosis, The result showed that miR-10a-5p bound to the 3ʹ-UTR of BDNF by a dual-luciferase reporter assay. Down-regulation of miR-10a-5p induced up-regulation of TrkB promoting development of laryngeal cancer. In vivo, down-regulation of TrkB suppressed tumor growth and inhibited the expression of stem-like marker proteins and promoted apoptosis.ConclusionIn conclusion, down-regulation of TrkB plays an important role in laryngeal cancer and is a promising target for future intervention strategies.

An Exploratory Study into the Implantation of Arytenoid Cartilage Scaffold in the Horse.

Respiratory function in the horse can be severely compromised by arytenoid chondritis, or arytenoid chondropathy, a pathologic condition leading to deformity and dysfunction of the affected cartilage. Current treatment in cases unresponsive to medical management is removal of the cartilage, which can improve the airway obstruction, but predisposes the patient to other complications like tracheal penetration of oropharyngeal content and dynamic collapse of the now unsupported soft tissue lateral to the cartilage. A tissue engineering approach to reconstructing the arytenoid cartilage would represent a significant advantage in the management of arytenoid chondritis. In this study, we explored if decellularized matrix could potentially be incorporated into the high motion environment of the arytenoid cartilages of horses. Equine arytenoid cartilages were decellularized and a portion of the resultant acellular scaffolds was implanted in a full-thickness defect created in the arytenoids of eight horses. The implantation was performed bilaterally in each horse, with one side randomly selected to receive an implant seeded with autologous bone marrow-derived nucleated cells (BMNCs). Arytenoids structure and function were monitored up to 4 months. In vivo assessments included laryngeal ultrasound, and laryngeal endoscopy at rest and during exercise on a high-speed treadmill. Histologic evaluation of the arytenoids was performed postmortem. Implantation of the cartilaginous graft had no adverse effect on laryngeal respiratory function or swallowing, despite induction of a transient granuloma on the medial aspect of the arytenoids. Ultrasonographic monitoring detected a postoperative increase in the thickness and cross-sectional area of the arytenoid body that receded faster in the arytenoids not seeded with BMNCs. The explanted tissue showed epithelialization of the mucosal surface, integration of the implant into the native arytenoid, with minimal adverse cellular reaction. Remodeling of the scaffold material was evident by 2 months after implantation. Preseeding the scaffold with BMNCs increased the rate of scaffold degradation and incorporation. Replacement of arytenoid portion with a tissue-engineered cartilaginous graft preseeded with BMNCs is surgically feasible in the horse, is well tolerated, and results in appropriate integration within the native tissue, also preventing laryngeal tissue collapse during exercise.

Aberrant allelic-switch of antisense lncRNA IRAIN may be an early diagnostic marker in laryngeal cancer.

Laryngeal carcinoma is a common head and neck malignancy, however, the molecular mechanism of the disease has not yet been elucidated. The present study aimed to investigate the role of IGF1R antisense imprinted non-protein coding RNA (IRAIN) long non-coding (lnc)RNA in laryngeal carcinoma. In total, specimens of healthy pharynx tissue from 6 healthy individuals, carcinoma tissue and paracancerous tissue from 37 patients with laryngeal carcinoma were used in this study. The single nucleotide polymorphism (SNP) rs8034564 was used to distinguish the two parental alleles of IRAIN. DNA and RNA were extracted from tissue specimens and the IRAIN allelic gene was sequenced. Reverse transcription-quantitative PCR was used to determine the expression levels of IRAIN and Insulin-like growth factor 1 receptor (IGF1R) in laryngeal carcinoma and paracancerous tissue. Bisulfite genomic sequencing was used to determine IRAIN promoter DNA methylation status in laryngeal carcinoma tissue. The expression of IRAIN was di-allelic in healthy pharynx tissue, laryngeal carcinoma tissue and paracancerous tissue. Moreover, IRAIN expression in laryngeal carcinoma tissue was lower compared with paracancerous tissue (P<0.05). IRAIN expression was not associated with age, histological type, tumor stage and grade and lymph node metastasis. IRAIN allelic expression imbalance was present in laryngeal carcinoma and paracancerous tissue, but not in healthy pharynx tissue. SNP analysis (rs8034564) indicated there was an allelic-switch of the two parental alleles. Furthermore, epigenetic analysis revealed no extensive DNA methylation of CpG islands in the IRAIN gene promoter of laryngeal carcinoma. Therefore, it was suggested that IRAIN allele was non-imprinted in laryngeal carcinoma and healthy pharynx tissue. It was also demonstrated that IRAIN may be a potential tumor suppressor in laryngeal carcinoma, and that DNA methylation is not involved in the regulation of IRAIN gene immobilization in laryngeal carcinoma tissue. Thus, detection of IRAIN allelic expression imbalance and aberrant allele-switch may serve as an early diagnostic marker of laryngeal carcinoma.

Experience with laryngeal reinnervation using nerve-muscle pedicle in pediatric patients

ObjectiveVocal fold paralysis (VFP) in adducted position remains a challenge for airway surgeons. Alternatives to tracheostomies such as lateralization, cordectomy, and posterior rib grafting disrupt the laryngeal tissue or framework and carry an increased risk of aspiration. Laryngeal reinnervation using nerve-muscle pedicle (NMP), carries the distinct advantage of preserving the larynx, sparing the recurrent laryngeal nerve, and obtaining an active VF abduction.The aim of this study was to evaluate the success and complications of laryngeal reinnervation using nerve-muscle pedicle (NMP) in pediatric patients presenting with dyspnea related to VFP in adducted position. MethodsIn this case series performed at a tertiary care referral center, review of medical records on all pediatric patients with VFP in adduction treated with laryngeal reinnervation using NMP between 1999 and 2017. Data were collected on the preoperative flexible laryngoscopy, suspension micro-laryngoscopy, and laryngeal electromyography as well as post-operative clinical assessment of the voice and airway.All patients underwent surgery consisting of the transfer of an innervated omohyoid muscle pedicle onto the paralyzed posterior cricoarytenoid muscle.The main outcomes measured were the clinical and fiberoptic laryngoscopic airway assessment monthly for the first 6 months, then at 12 months and annually thereafter. Clinical assessment included dyspnea evaluation based on a visual analog scale and voice assessment using the GRBAS scores. Complications from the treatment were also noted. These outcomes were determined before collection of data. Results16 cases were identified, with a mean age of 4 years. The recurrent laryngeal nerve paralysis was bilateral in 3 cases and unilateral in 13 cases. There were no peri or postoperative complications. After a mean follow-up of 7 years, vocal fold abduction was observed in 10 out of 16 cases and disappearance of paradoxical inspiratory adduction in 3 cases. Persistent dyspnea was noted in 7 cases (44%), and moderate dysphonia was present in 11 cases (69%). Finally, additional procedures were necessary in 2 patients (13%) to achieve the outcomes. ConclusionsLaryngeal reinnervation using NMP may be used in pediatric patients. This procedure, is safe and allows us to spare the recurrent laryngeal nerve while obtaining an active VF abduction in the majority of cases, and an improvement in breathing in most cases. Quality of evidence4.

MYCT1 inhibits the EMT and migration of laryngeal cancer cells via the SP1/miR-629-3p/ESRP2 pathway

MYCT1 has an inhibitory effect on the migration of laryngeal cancer cells, although the underlying molecular mechanism remains unknown. In this study, we aimed to explore the mechanism of MYCT1 in the epithelial-mesenchymal transition (EMT) and migration of laryngeal cancer cells. We found that MYCT1 significantly decreased the expression of miR-629-3p but increased the expression of ESRP2 in laryngeal cancer cells. The expression of miR-629-3p and ESRP2 in laryngeal cancer tissues showed significantly positive and negative correlations with patient metastasis, respectively. miR-629-3p was confirmed to repress the expression of ESRP2 by targeting its 3’UTR. SP1 was verified to be a direct transcription factor for miR-629-3p and a downstream target of MYCT1. Moreover, MYCT1 inhibited the EMT and migration of laryngeal cancer cells through the SP1/miR-629-3p/ESRP2 pathway. Taken together, our results establish a novel MYCT1 signaling pathway in the EMT and migration of laryngeal cancer cells, thus providing important insights for further studying the pathway in the diagnosis and treatment of laryngeal cancer.

Rol de los agentes físicos en la rehabilitación vocal: una revisión de la literatura

Los agentes físicos son elementos naturales o artificiales que se aplican para el tratamiento de determinados síntomas o patologías. En la rehabilitación vocal su estudio es un área emergente, donde las revisiones sistemáticas y los meta-análisis son escasos. Esto, muchas veces, dificulta la toma de decisiones y la correcta elección por parte del clínico. El objetivo de este trabajo fue analizar el rol de los agentes físicos en la rehabilitación vocal. Se realizó una revisión de la literatura a través de la búsqueda de artículos en las bases de datos PubMed, EBSCOHost y Scielo. Se establecieron criterios de elegibilidad según tipo, año y características de los estudios. Se evaluaron 603 artículos, de los cuales, luego del análisis de su título, abstract y del cumplimiento de los criterios de elegibilidad, se seleccionaron 16. Se entregan resultados en base a la cantidad de participantes, nivel de evidencia, tipo y configuración del agente físico, procedimientos e instrumentos de evaluación y beneficios obtenidos. Los agentes físicos de mayor utilización en la clínica vocal son la electroterapia (TENS y NMES) y la laserterapia. En general, estos actúan como coadyuvantes en la terapia vocal. La TENS reduce el dolor, la tensión laríngea y la percepción de voz apretada durante la fonación. La NMES beneficia la activación neuromuscular de las cuerdas vocales y el uso de láser permite la recuperación de los tejidos laríngeos posterior a tareas de sobrecarga.

RalB degradation by dihydroartemisinin induces autophagy and IFI16/caspase-1 inflammasome depression in the human laryngeal squamous cell carcinoma

BackgroundInterferon-inducible 16 (IFI16)/caspase-1 inflammasome activates and secretes IL-1β. However, it is still unclear whether the IFI16 inflammasome is involved in human laryngeal squamous cell carcinoma. Autophagy directly removed inflammasome components and limited early IL-1β production. RalB is required for the crosstalk between inflammasome and autophagy in macrophages. Dihydroartemisinin (DHA), the main derived ingredient of artemisinin, has a variety of biological activities. The mechanism of DHA in regulating the crosstalk between IFI16 inflammasome and autophagy by inhibiting RalB expression was analyzed in order to provide clues for new therapeutic methods in laryngeal cancer.MethodsThe expression of IFI16 was analyzed by Oncomine and GEPIA databases and detected by Western blot and immunohistochemistry. The relationship between IFI16 inflammasome and autophagy was investigated by transmission electron microscopy, immunofluorescence assay, etc. in Hep-2, Cal-27 and HeLa cells treated with DHA. The xenograft tumor of hep-2 cell in nude mice were used to assess the effect of DHA on laryngeal cancer.ResultsIt was reported for the first time in this study that IFI16 was overexpressed and positively correlated with caspase-1 in laryngeal carcinoma tissues. DHA significantly inhibited the activation of inflammasome and reduced IL-1β production in the microenvironment of Hep-2 cell xenograft tumor in nude mice. Mechanistically, we found that DHA degraded RalB, inhibited USP33 expression, and triggered autophagy. Meanwhile, enhanced autophagy can reduce the expression of RalB and USP33. Furthermore, DHA promotes autophagy, which suppresses the activation of IFI16/caspase-1 inflammasome and IL-1β production.ConclusionsTherefore, our findings demonstrate that DHA may act as a RalB inhibitor to regulate the crosstalk between autophagy and IFI16/caspase-1 inflammasome, which inhibits IL-1β production in tumor microenvironment.

The Association of Human Papillomavirus in Benign and Malignant Laryngeal Lesions-a Pilot Study.

Various etiological factors have been described in laryngeal carcinogenesis. Tobacco and alcohol play a major role in the development of laryngeal cancers. However, recently there has been an overshadowing of association of human papillomavirus infection in laryngeal cancers. The aim of this study is to assess the prevalence of human papillomavirus (HPV) in malignant laryngeal lesions. This is a case control study. We conducted this study using tumor tissue specimens from 30 laryngeal squamous cell carcinoma patients and benign laryngeal tissue specimens from 30 cancer-free controls. The specimens from the lesion were sent to histopathological analysis as well as DNA extraction (DNeasy® Tissue kit) and polymerase chain reaction for detection of HPV. Positive samples underwent sequencing to detect the HPV serotype. Statistical analysis was performed using SPSS software. All 30 benign laryngeal lesion specimens were negative for the HPV DNA, while 4 of the 30 (7.5%) squamous cell carcinoma lesions were positive. One was HPV 16 and another one HPV 11. Two positive cases were not able to be sequenced probably due to low viral load. Our study suggests that the proportion of laryngeal squamous cell carcinomas attributable to infection by HPV seems to be low. Further prospective studies should be conducted with a larger group of patients to confirm the role of HPV in laryngeal cancers especially in treatment response and survival.

Three-dimensional finite element modeling for evaluation of laryngomalacia severity in infants and children.

This study was performed to investigate the feasibility of using a three-dimensional (3D) finite element model for laryngomalacia severity assessment. We analyzed laryngeal computed tomography images of seven children with laryngomalacia using Mimics software. The gray threshold of different tissues was distinguishable, and a 3D visualization model and finite element model were constructed. The laryngeal structure parameters were defined. The peak von Mises stress (PVMS) value was obtained through laryngeal mechanical analysis. The PVMS values of the laryngeal soft tissue and cartilage scaffolds were independently correlated with disease severity. After stress loading the model, the relationship between laryngomalacia severity and the PVMS value was apparent. However, the PVMS value of laryngeal soft tissue was not correlated with laryngomalacia severity. This study established the efficacy of a finite element model to illustrate the morphological features of the laryngeal cavity in infants with laryngomalacia. However, further study is required before widespread application of 3D finite element modeling of laryngomalacia. PVMS values of the laryngeal cartilage scaffold might be useful for assessment of laryngomalacia severity. These findings support the notion that structural abnormalities of the laryngeal cartilage may manifest as quantifiable changes in stress variants of the supraglottic larynx.

Features of CO&lt;sub&gt;2&lt;/sub&gt;-laser application in surgical treatment of chronic cicatrical stenoses of the larynx

The aim of the review is to identify current possibilities of carbon dioxide laser application in microsurgical treatment of chronic laryngeal cicatricial stenoses. Currently, laser endolaryngeal surgeries are one of the main standards in surgical treatment of patients with this laryngeal pathology, including when providing high-tech assistance. Various types of lasers and methods of surgical treatment of the larynx are used. The choice of these methods is primarily determined by the opinion of the operating surgeon. The review highlights the principles of laser surgery in general, and when using carbon dioxide laser in particular. The use of micromanipulators makes it possible to combine the work of carbon dioxide laser with the operating microscope, affecting target tissues noninvasively with maximum precision. This technology makes it possible to expand the range of surgical manipulations in endolaryngeal surgeries. The main operating modes of the carbon dioxide laser, applicable for different clinical situations, such as power, impulse, laser power density (PD) are considered. In addition, the combination of a micromanipulator with an operating microscope allows the use of additional techniques aimed at preventing restenosis, such as the use of microflaps, submucosal laryngeal cicatricial tissue vaporization. The possibilities of the recurrence of cicatricial stenosis by different variants of stenting are presented. The risk of complications of this technique is not higher than in other types of larynx laser surgeries, and with proper preventive measures can be minimized. In the review, possible complications, both early and late, measures of their prophylaxis, as well as ways to eliminate their consequences are analyzed in detail.

Diagnosis of laryngeal tuberculosis in a high TB burden area.

The larynx is the second most commonly affected site in the head and neck region in patients with extrapulmonary tuberculosis (TB). Despite this, the prevalence of laryngeal TB is largely unknown, particularly in areas with a high TB burden. The laboratory diagnosis of TB includes microscopy, culture and molecular testing. The aims of this study were to determine the prevalence of laryngeal TB in patients presenting with laryngeal pathology in a region with a high TB burden and to determine the optimal diagnostic methods for the diagnosis of laryngeal TB. This was a prospective descriptive study of 80 adult patients undergoing direct laryngoscopy and biopsy for laryngeal pathology in the Department of Otorhinolaryngology, Universitas Academic Hospital, Bloemfontein, South Africa over a 1year period. Histopathological and microbiological investigations (microscopy, Xpert MTB/RIF, and TB culture) were performed on all laryngeal biopsies. Five (6.25%) out of 80 patients were diagnosed with laryngeal TB. In one patient, the Xpert MTB/RIF assay was positive on the laryngeal tissue and histology showed granulomas. Two patients had granulomas on histology although the microbiological tests on the tissue were negative. Two patients had only positive tissue cultures for Mycobacterium tuberculosis. None of the biopsies had positive Ziehl-Neelsen stains. The results suggest that the diagnosis of laryngeal TB required a combination of histopathology, culture and PCR and that the Xpert MTB/RIF assay is not a sensitive test for the diagnosis of laryngeal TB.

FADS1 promotes the progression of laryngeal squamous cell carcinoma through activating AKT/mTOR signaling

Metabolic abnormality is the major feature of laryngeal squamous cell carcinoma (LSCC), however, the underlying mechanism remain largely elusive. Fatty acid desaturase 1 (FADS1), as the key rate-limiting enzyme of polyunsaturated fatty acids (PUFAs), catalyzes dihomo-gamma-linolenic acid (DGLA) to arachidonic acid (AA). In this study, we reported that the expression of FADS1 was upregulated in LSCC, high FADS1 expression was closely associated with the advanced clinical features and poor prognosis of the recurrent LSCC patients after chemotherapy. Liquid chromatograph-mass spectrometry (LC-MS) analysis revealed that FADS1 overexpression induced greater conversion of DGLA to AA, suggesting an increased activity of FADS1. Similarly, the level of prostaglandin E2 (PGE2), a downstream metabolite of AA, was also elevated in cancerous laryngeal tissues. Functional assays showed that FADS1 knockdown suppressed the proliferation, migration and invasion of LSCC cells, while FADS1 overexpression had the opposite effects. Bioinformatic analysis based on microarray data found that FADS1 could activate AKT/mTOR signaling. This hypothesis was further validated by both in vivo and in vitro assays. Hence, our data has supported the viewpoint that FADS1 is a potential promoter in LSCC progression, and has laid the foundation for further functional research on the PUFA dietary supplementation interventions targeting FADS1/AKT/mTOR pathway for LSCC prevention and treatment.

LncRNA HCG11 suppresses laryngeal carcinoma cells progression via sponging miR-4469/APOM axis.

Longnon-coding RNAs (lncRNAs) have been reported to participate in the regulatory mechanisms of various cancers. Therefore, the aim of this study was to investigate the functional role of lncRNA HLA complex group 11 (HCG11) in laryngeal carcinoma. The laryngeal carcinoma cell lines SNU46, SNU899, AMC-HN-8, and normal human nasopharyngeal epithelial cells NP69 were purchased. The expression of HCG11, miR-4469, and apolipoprotein M (APOM) was detected by quantitative Real Time-PCR (qRT-PCR) in tissues and cells. Cell proliferation was detected by Cell Counting Kit-8 (CCK-8) assay and colony formation assays. The protein expression of Bax and Bcl-2 was detected by Western blot. Besides, the mechanism assays were conducted to observe the interaction between miR-449 and HCG11 or APOM. The apoptosis in each group was detected by TUNEL assay. In this research, low expression of HCG11 was discovered in laryngeal carcinoma tissues and cells. Overexpression of HCG11 retarded cell proliferation and enhanced cell apoptosis. Later, we found that APOM was also downregulated in laryngeal carcinoma tissues and cell lines, and inhibited laryngeal carcinoma progression. HCG11 positively regulated APOM at the post-transcriptional level. MiR-4469 was predicted to have the binding sites of HCG11 and APOM. Furthermore, it was demonstrated that HCG11 absorbed miR-4469 to upregulate APOM expression. Finally, it was indicated that the repression of APOM rescued the effects of HCG11 overexpression on cell proliferation and cell apoptosis. This study uncovered that HCG11 sponged miR-4469 to suppress laryngeal carcinoma progression by upregulating APOM expression.

Properties of a Novel Animal Model of LPR

Few satisfactory animal models of laryngopharyngeal reflux (LPR) is available. Interleukin-8 (IL-8) and vascular endothelial growth factor (VEGF) may be associated with the pathogenesis of LPR injuries and laryngeal carcinomas. To establish an animal model of LPR and to explore the related pathological changes and cytokine expression in the vocal cord tissue. Twenty rabbits were divided into experimental and control groups. Dilatation of the upper and lower esophageal sphincter were carried out in the experimental group. The pH of the pharynx, pathological, and ultrastructural changes of the laryngeal tissue, and expression of IL-8 and VEGF were compared between the experimental group and controls. pH monitoring results and the dilated intercellular space of the vocal cord mucosa showed that the experimental group developed laryngopharyngeal reflux. There were significant differences in the immunohistochemical staining scores of both IL-8 (P=0.015) and VEGF (P=0.007) between the experimental and control groups in the vocal cord tissue. We successfully established a model of LPR, showing histopathological and ultrastructural changes consistent with the disease. The expression of IL-8 and VEGF may increase during the pathogenesis of LPR.