Search

Objectives/hypothesis: Vocal fold epithelium is exposed to reactive oxygen species from the inhaled environment and from tissue inflammation. The objective of this study was to explore the functional and structural consequences of reactive oxygen species exposure on vocal fold epithelium., Study Design: In vitro, prospective study design., Methods: Hydrogen peroxide (H(2)O(2)), a common reactive oxygen species, was utilized in this study. Freshly excised, viable porcine vocal fold epithelia (N = 32) were exposed to H(2) O(2) or sham challenge for 2 hours. Electrophysiology, western blotting, and light microscopy were used to quantify the functional and structural effects of reactive oxygen species on vocal fold epithelia., Results: Exposure to reactive oxygen species did not significantly alter transepithelial resistance. There was a small, nonsignificant trend for decreased concentration of epithelial junctional complex protein with reactive oxygen species challenge. Minimal changes to the gross structural appearance of vocal fold epithelia were also noted., Conclusions: The stratified squamous epithelia of the vocal folds effectively defend against an acute reactive oxygen species challenge. The current study lays the groundwork for future investigations on the effects of reactive oxygen species on vocal fold epithelia that are compromised from phonotrauma., (Copyright © 2011 The American Laryngological, Rhinological, and Otological Society, Inc.)

The expression of angiogenesis-related proteins was determined in laryngeal tumour tissue, associated tumour involved lymph nodes, apparently normal mucosa and control tissue and were related to tumour stage. Both laryngeal tumour tissue and associated metastatic nodes were obtained from seven patients undergoing surgical resection; in four cases apparently normal mucosa was also dissected from the tumour specimen margins. Control uvula mucosa was obtained from five healthy volunteers undergoing uvulopalatopharyngoplasty. The relative expression of 55 angiogenesis-related proteins was determined in tissue lysates using a Proteome Profiler human angiogenesis array kit. The level of 32/55 angiogenesis-related proteins was higher in tumour tissue compared with controls. Furthermore, in these tumour biopsies higher levels of proteins were associated with increasing tumour stage. A similar trend was seen for 29/32 of these proteins in the nodal tissue. In T4 stage tumour tissue samples, 29/55 angiogenensis-related proteins were more highly expressed compared with the adjacent normal mucosa from the same patient, and this decreased to 8 proteins in tumour tissue from the T1 stage patients. In contrast, the expression of 23 angiogenesis-related proteins in metastatic lymph node tissue from T4 stage patients was lower compared with that found in the normal mucosa adjacent to the tumour. In conclusion, this study has identified a number of factors involved in angiogenesis that are likely to contribute to the growth and metastasis of laryngeal tumours. Furthermore, a number of factors were also substantially altered in metastatic deposits compared with the primary tumour mass or adjacent normal tissue. This study requires confirmatory analysis of the selected key factors in a larger cohort of patients.

Objectives/hypothesis: To investigate whether the pepsin immunohistochemical (IHC) staining of the laryngeal mucosa epithelia is an available test for diagnosing laryngopharyngeal reflux (LPR) in clinic., Study Design: Prospective case series., Methods: Biopsy specimens from interarytenoid mucosa of LPR patients (seven acid LPR and eight nonacid LPR) and 21 sex- and age-matched normal controls were obtained for pepsin IHC staining. The diagnosis of LPR was based on 24-hour combined multichannel intraluminal esophageal impedance pH monitoring. The results of IHC staining were semiquantitatively analyzed and scored as negative (-), weakly positive (+), moderately positive (++), and strongly positive (+++)., Results: Six of seven acid LPR (85.7%) and six of eight nonacid LPR (75%) mucosa samples were moderate to strongly positive for intracellular pepsin. By contrast, only three of 21 normal controls (14.3%) were moderately positive. The difference in intracellular pepsin between LPR and the normal laryngeal mucosa was statistically significant (P < .01). No significant difference in intracellular pepsin was observed between the acid and nonacid LPR mucosal samples (P = .453). Using weak positivity (+) as a cutpoint, the presence of intracellular pepsin in the laryngeal mucosa had a sensitivity of 100% and a specificity of 47.6% in detecting LPR (P < .05). However, using the moderate positivity (++) as the cutpoint, the pepsin had a slightly decreased sensitivity of 80% but a sharply increased specificity of 85.7% (P < .05) in the detection of LPR., Conclusions: Pepsin IHC staining of the laryngeal mucosa appears to be a sensitive and specific test for diagnosing LPR in a clinical application., (Copyright © 2011 The American Laryngological, Rhinological, and Otological Society, Inc.)

Objective: To determine the contribution of B- and T-cell-mediated inflammation in a murine airway granulation model., Study Design: Pilot study in a modified murine model., Setting: Philadelphia VA Medical Center Research Building., Subjects and Methods: Laryngotracheal complexes (LTCs) from 54 donor C57BL/6 mice were harvested and divided into 3 groups: (1) uninjured, (2) mechanically injured using a wire brush, and (3) chemically injured using hydrochloric acid. One donor LTC from each group was placed in deep dorsal subcutaneous pockets of either severe combined immunodeficiency (SCID)- or C57BL-recipient mice, for a total of 3 transplanted tracheas per recipient mouse. After 3 weeks, the transplanted LTCs were harvested from both C57BL- and SCID-recipient mice. Tissues were fixed, sectioned, and stained with hematoxylin and eosin. Representative slides were reviewed by a blinded pathologist to determine the formation of granulation tissue and graded as to the degree of formation of granulation tissue., Results: Despite significant granulation formation in C57BL-recipient mice, direct airway injury did not induce the formation of granulation tissue under the disrupted epithelium of airway mucosa in SCID mice 3 weeks after injury., Conclusion: The data indicate that the immune response that results in the formation of granulation tissue is mediated by circulating B- and/or T-cell processes rather than resident airway immune cells. Further studies focusing on cellular adaptive immune processes in response to airway injury may provide a novel treatment modality for subglottic stenosis.

Background: To investigate the pathophysiology of reflux laryngitis, an experimental model is required., Aim: The aim of this study is to establish an animal model of reflux esophago-laryngitis, modifying our previously reported model of chronic acid reflux esophagitis., Methods: The modified chronic acid reflux esophagitis (m-RE) group (n = 10), in which the duodenum was wrapped with 2.5 mm of Nelaton catheter, was not treated with any drugs. Also postoperatively, two treatment groups (n = 10 in each) received different dosages of rabeprazole (RPZ): 1.0 mg/kg/day (RPZ 1.0 group) or 10.0 mg/kg/day (RPZ 10.0 group). As a control group (n = 5), other rats underwent sham operation. The esophagus and larynx were resected on day 14 after the operation, and ulcer score of the esophagus was assessed. The epithelial thickness and leukocyte infiltration of the supraglottic and subglottic laryngeal mucosae were investigated. The number of interleukin (IL)-1β-positive cells was also counted and defined as the IL-1β labeling index., Results: In the m-RE group, the epithelial thickness, leukocyte infiltration, and IL-1β labeling index of the supraglottic and subglottic laryngeal mucosae were increased compared with controls (P < 0.01). In the RPZ groups, not only the ulcer score of esophagus but also the epithelial thickness, leukocyte infiltration, and IL-1β labeling index of both the supraglottic and subglottic laryngeal mucosae were decreased dose-dependently relative to the m-RE group (P < 0.05)., Conclusions: Our modified chronic acid reflux esophagitis model proved useful in establishing a rat reflux esophago-laryngitis model, with both pathological laryngeal findings and reflux esophagitis shown to be improved by administration of a proton pump inhibitor.

Objectives: To report use of a shielded hook bistoury developed for correction of epiglottic fold entrapment in standing sedated horses., Study Design: Prospective clinical study., Animals: Standardbreds (n=8), 3-19 years of age., Methods: Epiglottic fold entrapment was corrected during standing sedation using a shielded hook bistoury passed nasally into the pharynx. Owners, trainers, and referring veterinarians were contacted by telephone to establish outcome., Results: Axial division of the entrapping epiglottic fold was successful in all horses. No short- or long-term complications occurred. Axial transection of the entrapping fold was performed rapidly (mean, 83 seconds between advancing the bistoury into the pharynx and transection of the entrapping fold)., Conclusion: Transnasal axial division of the aryepiglottic fold can be accomplished safely using the shielded hook bistoury. The shield mechanism minimizes the risk of injury from the cutting blade to adjacent structures., (© Copyright 2010 by The American College of Veterinary Surgeons.)

Objective: Supracricoid laryngectomy (SCL) is a reliable procedure for laryngeal preservation. However, due to the limited anatomy of the larynx, close surgical margins are often inevitable. Although the usefulness of intraoperative margin study on frozen section has been well documented in laryngeal surgery, the clinical significance of margin study in SCL has not been clarified. This study evaluated the evidence base supporting margin study in SCL., Methods: Between 1997 and 2009, 61 patients underwent SCL. Margin study was conducted by histopathologically examining surrounding mucosal strips between the resected laryngeal specimen and the residual larynx using frozen sections. The findings were analyzed in terms of pT staging and prognoses., Results: Among all patients, pathological report indicated all negative in 36, dysplasia in 18, and positive findings in seven patients. Positive results were exclusively identified at the ipsilateral posterior and inferior margins. The incidence of local recurrence and death due to disease was slightly higher in patients with positive reports. The margin study influenced the intraoperative decision to convert from SCL to total laryngectomy in one case., Conclusions: In reviewing the margin study of 61 SCL patients, 11% resulted in positive margin. All except one patient with positive margin attained negative finding with additional samplings. Decision making regarding the resection margin can be difficult in patients with pT3-pT4 stages and postradiation status. Because of the exclusive identification of positive margin at the ipsilateral posterior and inferior edges, the margin study is strongly recommended at these sites. The possibility of converting SCL to TL should be discussed preoperatively during the informed consent process. The margin study is an effective modality for ensuring the validity of SCL and is recommended for all SCL procedures., (Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.)

We present the case of a 41-year-old man with a herniated saccular laryngeal cyst in the left cervical region who died unexpectedly at home from acute asphyxia. A few days before death the patient complained an obvious palpable swelling on the left side of the neck. The autopsy showed an oval-shaped mass originating on the wall of the larynx and herniating into the extralaryngeal region. The critical reduction of the laryngeal lumen originated from an intense oedema of the laryngeal mucosa above the cyst, concurrent with an inflammatory process of the cystic wall, with probable associated glottis laryngospasm mediated by the superior laryngeal nerve. The rapid evolution of oedema was favoured by the extreme relaxation of the connective tissue of the laryngeal mucosa in the supraglottic and aryepiglottic regions which explains the sudden death. The case is useful for delineating the problems related to the possible evolution of laryngeal cysts, the mechanisms of asphyxial complications, the pathological diagnosis and the medico-forensic aspects., (Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.)

The mechanisms leading to death from anaphylaxis after insect sting involve antigen cross-linkage of antibody molecules that activate immunoglobulin receptors on inflammatory cells. The aim of our study was to investigate the pathomorphology and the expression of C5aR in fatal anaphylaxis in a patient after a fatal insect sting. A 38-year-old women was stung by a honeybee. C5R1 expression was detected in many dilated capillaries in the lungs. Pulmonary epithelial cells did not bind the monoclonal antibody for C5R1; however, intensive cytoplasmic staining was detected in endothelial cells. The findings of this case provide evidence for an active role of complement in fatal anaphylaxis elicited by bee venom. C5aR detection could be useful in the identification of sudden death cases because of unwitnessed fatal insect sting cases. Authors can recommend this immunohistochemical analysis on all sudden unexpected deaths outdoors where a possible bee sting might occur., (© 2011 American Academy of Forensic Sciences.)

Objectives/hypothesis: To determine the neuropathologic findings in tissue obtained from children with laryngomalacia at a tertiary-care pediatric hospital., Study Design: Retrospective review of consecutive cohort compared with a control group., Methods: We reviewed supra-arytenoid pathology specimens from 43 children with severe laryngomalacia and 13 age-matched pediatric autopsy controls. Histopathologic comparison was made of nerve hypertrophy (including nerve perimeter and surface area) among experimental and control pathologic specimens., Results: There exists a statistically significant increase in nerve perimeter (P = .001) and nerve surface area (P = .02) in supra-arytenoid specimens in patients with severe laryngomalacia compared with age-matched autopsy supra-arytenoid tissue., Conclusions: The pathologic finding of nerve hypertrophy in children with laryngomalacia provides new evidence to support neurologic dysfunction as the etiologic theory of laryngomalacia., (Copyright © 2011 The American Laryngological, Rhinological, and Otological Society, Inc.)

Objective: To investigate the characteristics of the laryngeal mucosal microvascular network in suspected laryngeal cancer patients, using narrow band imaging, and to evaluate the value of narrow band imaging endoscopy in the early diagnosis of laryngeal precancerous and cancerous lesions., Patients and Methods: Eighty-five consecutive patients with suspected precancerous or cancerous laryngeal lesions were enrolled in the study. Endoscopic narrow band imaging findings were classified into five types (I to V) according to the features of the mucosal intraepithelial papillary capillary loops assessed., Results: A total of 104 lesions (45 malignancies and 59 nonmalignancies) was detected under white light and narrow band imaging modes. The sensitivity and specificity of narrow band imaging in detecting malignant lesions were 88.9 and 93.2 per cent, respectively. The intraepithelial papillary capillary loop classification, as determined by narrow band imaging, was closely associated with the laryngeal lesions' histological findings. Type I to IV lesions were considered nonmalignant and type V lesions malignant. For type Va lesions, the sensitivity and specificity of narrow band imaging in detecting severe dysplasia or carcinoma in situ were 100 and 79.5 per cent, respectively. In patients with type Vb and Vc lesions, the sensitivity and specificity of narrow band imaging in detecting invasive carcinoma were 83.8 and 100 per cent, respectively., Conclusion: Narrow band imaging is a promising approach enabling in vivo differentiation of nonmalignant from malignant laryngeal lesions by evaluating the morphology of mucosal capillaries. These results suggest endoscopic narrow band imaging may be useful in the early detection of laryngeal cancer and precancerous lesions.

Background: The aim of this study was to analyze genetic alterations in the transformation-progression model of laryngeal tumors., Methods: Copy number changes of 37 genes were analyzed by multiple ligation-dependent probe amplification (MLPA) in 94 tissue samples., Results: In the smoker normal mucosa group TP53 loss was predominant, whereas in the precursor lesions CDKN2A loss and CDKN2D gain were most frequent. Precursor lesions with progression presented CTNNB1 loss. In the carcinoma group the most common changes were CDKN2A, MLH1, CTNNB1, and CASP6 losses and RECQL4, CCND1, and EMS1 gains. Positive lymph node primary tumors were related to TP53, IL1A, and RB1 losses and STK11 gain. The lymph node metastases differed from their corresponding primary tumor in LMNA, RECQL4, and IGF1R losses, and N33 and CDKN2D gains., Conclusions: Genetic changes and new key genes were found to be associated with specific steps. We included new steps, not presented in the classic models: normal mucosa tobacco exposed, positive lymph node primary tumor, and corresponding lymph node metastases., (Copyright © 2010 Wiley Periodicals, Inc.)

Laryngeal manifestations due to estrogen deficiency have been studied in the literature. But to date, the possible histopathological changes of laryngeal mucosa due to estrogen deficiency have not been studied. Therefore, our objective was to determine the histopathological changes of laryngeal mucosa in ovariectomised rats in order to clarify effects of estrogen deficiency on laryngeal tissue. The study is a randomized trial and was conducted at the animal care facility of Haydarpasa Numune Education and Research Hospital. Twenty-one Wistar rats were used throughout the experiment. There were six rats in the sham-operated control group. And others were divided into two groups (4, 8 weeks) according to follow-up time after ovariectomy. We observed significant changes 4 weeks after ovariectomy when we assessed subepithelial edema, inflammation, cilia and goblet cell loss (p < 0.01). It was shown that estrogen deficiency after ovariectomy in rats caused changes in laryngeal tissue when it was studied histopathologically.

Purpose: The aim of the study was to provide a morphological assessment of the laryngeal mucosa in patients with hyperfunctional dysphonia diagnosed by psychoacoustic and videostroboscopic methods., Material and Methods: Forty patients with voice quality disorders of hyperfunctional dysphonia were recruited for participation in the study. The diagnosis of dysphonia was based on the Voice Rating Scale GRBAS, and endoscopic and stroboscopic assessment of the vocal folds. Acoustic assessment was carried out using following parameters: fundamental frequency, Jitter, Shimmer, Noise to Harmonic Rate and Yanagihara (YG) scale. In order to evaluate the morphology of the vocal fold mucosa transmission electron microscopy was performed using postoperative material obtained from the larynx. Results of clinical and morphological analysis were compared with the reference group. The morphological material was obtained from patients with hypopharyngeal cancer without pathological changes of the vocal folds., Results: The psychoacoustic assessment using the perceptual GRBAS scale enables the appropriate diagnostics of hyperfunctional dysphonia, which was confirmed by evaluation of acoustic parameters and YG scale analysis. In 40 patients with voice quality disorders causing by hyperfunctional dysphonia, in morphological assessment of the laryngeal mucosa, 4 (10%) patients demonstrated the presence of oedema and signs of intensive dysphonia in psychoacoustic and stroboscopic examination., Conclusions: Oedema of the laryngeal mucosa confirmed by stroboscopic and ultramorphological examination may coexist with hyperfunctional dysphonia. The presence of the laryngeal oedema in patients with hyperfunctional dysphonia has the negative impact on voice quality in psychoacoustic assessment with the use of the GRBAS and YG scales.

The postmortem diagnosis of anaphylactic death may be frustrating when victims are not hospital patients, even more so when they are recovered dead. The frequent lack of specific morphological findings in such cases means that diagnosis by the forensic pathologist must rely solely on exclusion criteria or circumstantial evidence. However, a diagnostic approach based on case history, analysis of circumstances, available clinical and necropsy findings, as well as toxicology, histopathology and biohumoral data, often allows demonstration of the cause of death. Some useful reflections on microscopic morphological data have come from two recent cases, where thorough data collection provided a reasonably certain diagnosis of anaphylactic death and systemic inflammatory response syndrome-related cardiac arrest, respectively. In both cases tissue histopathology proved crucial, since histochemical (GIEMSA) and immunohistochemical analysis (CD117 and tryptase) documented a large number of mast cells in tissues, particularly the laryngeal wall, and a discrepancy between cells positive for GIEMSA and tryptase and those positive for CD117. Staining for CD117 was also detected in cells with dendrite morphology and in a subpopulation of small lymphocytes with incised nuclei. The morphological findings of these cases are discussed, especially those obtained with immunohistochemistry, and the need for the latter data to be interpreted by experienced medical staff in the framework of a thorough analysis of all the data collected is highlighted.

Vocal folds play a crucial role in voice production. The physiological vibrations of vocal folds depend on the unchanged multilayered structure of the vocal folds mucosa. Morphological changes of mucosa are the cause of voice quality disorders - dysphonia. The aim of this study was to determine the morphological base of dysphonia in patients with vocal folds atrophy. A group of 24 patients with larynx atrophy confirmed by endoscopic (VLS) and stroboscopic (VLSS) examination of the larynx was included in the study. The morphological assessment of the larynx mucosa was carried out with the use of the transmission electron microscopy (TEM). Ultramorphological examinations revealed changes in the epithelium, basal membrane and lamina propria of the vocal folds mucosa. An increased number of collagenous fibers, fibroblasts with signs of vacuolar degeneration inflammatory cells and a decreased number of blood vessels and pericytes were observed. Morphological changes found in the epithelium, basal membrane and lamina propria of the vocal folds mucosa were the cause of disorders of vocal folds vibrations registered in the stroboscopic examination of the larynx (VLSS).

Objective: To assess whether dynamically modulating endotracheal tube (ETT) cuff pressure, by decreasing it during each ventilatory cycle instead of maintaining a constant level, would reduce the extent of intubation-related laryngotracheal injury., Design: Single-blind, randomized controlled animal study using a previously validated live porcine model of accelerated intubation-related tracheal injury., Setting: Animal research facility., Patients: Ten piglets (weight, 16-20 kg each) were anesthetized and underwent intubation using a cuffed ETT., Interventions: The animals were randomized into the following 2 groups: 5 pigs had a novel device to modulate their cuff pressure from 25 cm H₂O during inspiration to 7 cm H₂O during expiration, and 5 pigs had a constant cuff pressure of 25 cm H₂O. Both groups underwent ventilation under hypoxic conditions for 4 hours., Main Outcome Measure: Laryngotracheal mucosal injury after blinded histopathological assessment., Results: The modulated-pressure group showed significantly less overall laryngotracheal damage than the constant-pressure group (mean grades, 1.2 vs 2.1; P < .001). Subglottic damage and tracheal damage were significantly less severe in the modulated-pressure group (mean grades, 1.0 vs 2.2; P < .001, and 1.9 vs 3.2; P < .001, respectively). There was no significant difference in glottic or supraglottic damage between the groups (P = .06 and .27, respectively)., Conclusions: This novel device reduces the risk of subglottic and tracheal injury by modulating ETT cuff pressure in synchronization with the ventilatory cycle. This finding could have far-reaching implications for reducing the risk of airway injury in patients undergoing long-term intubation. Further clinical study of this device is warranted.

Objective: To investigate the histopathologic changes in intact laryngeal epithelium and mucosa exposed to endogenous gastric acid and pepsin in an experimental model of reflux., Study Design: Randomized trial., Setting: The study was conducted at the animal care facility of Haydarpasa Numune Education and Research Hospital., Subjects and Methods: Eighteen healthy 200- to 220-g, 20-week-old Sprague-Dawley rats were used. The animals were divided into three groups according to exposure time (1-, 4-, and 12-week exposures), and four rats were examined as controls who underwent sham operation. An experimental model of gastroesophageal reflux was induced. After exposure, the animals were euthanized, and their larynges were removed. The histopathologic changes in the larynx were observed under a light microscope., Results: The mean scores for inflammation in the control, one-, four-, and 12-week groups were 0.75 ± 0.50, 1.75 ± 0.50, 2.20 ± 0.45, and 1.94 ± 0.87, respectively. However, mean scores for vascular engorgement in the control, one-, and four-week groups were 0, and in the 12-week group was 2.0 ± 0.70. The mean scores for subepithelial edema in the control, one-, four-, and 12-week groups were 1.00 ± 0, 1.75 ± 0.95, 1.80 ± 0.45, and 2.20 ± 0.84, respectively. However, mean scores for keratinization for the control, one-, and four-week groups were 0, and for the 12-week group was 1.60 ± 0.55. When we compared inflammation, vascular engorgement, subepithelial edema, and keratinization mean scores between the control and study groups, there were statistically significant increases (P = 0.005, P = 0.001, P = 0.043, P = 0.002, respectively)., Conclusion: Our findings suggest that reflux induces significant histopathologic changes in larynx mucosa., (Copyright © 2010 American Academy of Otolaryngology-Head and Neck Surgery Foundation. Published by Mosby, Inc. All rights reserved.)

Purpose of Review: To review recent literature on animal models used to study the pathogenesis, detection, prevention, and treatment of vocal fold scarring. Animal work is critical to studying vocal fold scarring because it is the only way to conduct systematic research on the biomechanical properties of the layered structure of the vocal fold lamina propria, and therefore develop reliable prevention and treatment strategies for this complex clinical problem., Recent Findings: During the period of review, critical anatomic, physiologic, and wound healing characteristics, which may serve as the bases for selection of a certain species to help answer a specific question, have been described in mouse, rat, rabbit, ferret, and canine models. A number of different strategies for prophylaxis and chronic scar treatment in animals show promise for clinical application. The pathways of scar formation and methods for quantifying treatment-induced change have become better defined., Summary: Recent animal vocal fold scarring studies have enriched and confirmed earlier work indicating that restoring pliability to the scarred vocal fold mucosa is challenging but achievable. Differences between animal models and differences in outcome measurements across studies necessitate considering each study individually to obtain guidance for future research. With increased standardization of measurement techniques it may be possible to make more inter-study comparisons.

Purpose of Review: To present the accurate surgical indication for the slicing mucosal technique, the case selection, surgical aspects, rehabilitation concerns, and the characteristics of immediate and long-term outcomes., Recent Findings: The literature is still scarce; few cases are submitted to the slicing mucosa technique due to its specific indication; an alternative procedure was designed for cases where mucosal movement is strongly reduced, the inner section of the vocal ligament or submucosal scar tissue, which can eventually be associated with fat inclusion. Some selected cases may require thyroplasty type III to optimize functional results., Summary: Slicing technique is an aggressive powerful resource for the surgical treatment of severe cases of sulcus striae major, in which mucosal wave is absent and glottic chink is moderate to severe; voice is intensely deviated immediately postoperation; vocal rehabilitation is mandatory and an intensive regimen is usually required for the first 2 months; final results can mostly be achieved up to 6 months.

Objective: Airway compromise due to paediatric intubation injuries is well documented; however, intubation injuries may also cause severe voice disorders. We report our experience and review the world literature on the voice effects of traumatic paediatric intubation., Case Series: We report five cases of children referred to Great Ormond Street Hospital for Children who suffered traumatic avulsion of the vocal fold at the time of, or secondary to, endotracheal intubation. All children had significant dysphonia and underwent specialist voice therapy., Conclusions: The mechanisms of injury, risk factors and management of the condition are discussed. Children suffering traumatic intubation require follow up throughout childhood and beyond puberty as their vocal needs and abilities change. At the time of writing, none of the reported patients had yet undergone reconstructive or medialisation surgery. However, regular specialist voice therapy evaluation is recommended for such patients, with consideration of phonosurgical techniques including injection laryngoplasty or thyroplasty.

Background: Overexpression of the lysyl oxidase-like 4 (LOXL4) gene is associated with a variety of human malignancies. The purpose of this study was to compare the gene expression of LOXL4 to the expression of epidermal growth factor receptor (EGFR) in head and neck squamous cell carcinomas. The overexpression of EGFR has been well examined and already serves as a therapeutic target. The expression of both genes was compared in head and neck squamous cell carcinomas and their diagnostic and prognostic value was evaluated., Materials and Methods: Messenger RNA from 58 head and neck carcinomas and 11 healthy upper aerodigestive tract mucosa samples was extracted and subjected to electrophoresis. Northern hybridisation was carried out using digoxigenin-labelled gene-specific probes, and the level of gene expression was measured by densitometry., Results: High expression of LOXL4 gene was detected in 71% of all carcinomas and only in 9% of the healthy mucosa samples (p=0.0002). In comparison, a high level of expression was detected for the EGFR gene in 78% of the carcinomas and in 36% of normal mucosa (p=0.01)., Conclusion: Although both genes revealed a similar level of overexpression in the carcinoma samples, it was found that the a notably higher percentage of healthy mucosa tested positively for EGFR than LOXL4, indicating that LOXL4 may serve as a selective molecular marker in primary and metastatic head and neck carcinoma.

Failure to remove tumor cells from the larynx significantly increases the risk of local recurrence following surgical excision. Healthy tissue must be preserved to optimize long-term vocal and swallowing function. It is essential to accurately distinguish between healthy mucosa, dysplasia, and invasive carcinoma. Optical and molecular examining technologies have been developed to improve tumor margin identification in vivo. We aimed to review the efficacy of these technologies. Published articles were identified using MEDLINE, EMBASE, and Cochrane central register of controlled trials (CENTRAL). Randomized clinical trials are required to establish the benefit to patients and cost to the health service of using 5-aminolevulinic acid (ALA)-induced fluorescent imaging, contact endoscopy, and optical coherence tomography (OCT). Furthermore, primary research is required to validate other techniques, such as confocal endomicroscopy and Raman spectroscopy, and to develop their clinical applications in the larynx.

Primary non-Hodgkin lymphoma of the larynx is rare, accounting for less than 1% of all laryngeal neoplasms. Fewer than 100 cases have been reported in the literature. This case describes a 76-year-old woman with primary non-Hodgkin lymphoma of the larynx. To our knowledge, this is the first case of primary non-Hodgkin lymphoma of the larynx reported in the Danish literature.

Mucosal leishmaniasis is uncommon outside Central and South America, where it is commonly caused by Leishmania (L.) braziliensis. We present a case of isolated laryngeal leishmaniasis detected in a 78-year-old male, who presented with chronic hoarseness. Histologic examination of biopsies taken from the larynx showed L. amastigotes. An L.-specific indirect fluorescent antibody test was positive. Polymerase chain reaction showed infection with L. donovani, L. infantum or L. tropica, species which do not normally cause isolated mucosal infection. This is the first reported case from Scandinavia.

Objectives: We describe the distribution of elastic system fibers in the superficial layer of the lamina propria of Reinke's edema, as compared with normal vocal folds., Methods: Weigert's resorcin-fuchsin stain after oxidation with 10% oxone was used to study the arrangement of elastic fibers. The findings were categorized and afterward compared with the severity of Reinke's edema., Results: Analysis of 20 specimens of vocal folds with Reinke's edema showed that the network of thin elastic fibers in Reinke's space lost their undulated appearance and had a tangled aspect. In addition, these fibers were no longer parallel to the epithelial basement membrane, but had a random distribution scattered throughout Reinke's space. The elastic fiber network immediately below the epithelial basement membrane also appeared more fragmented in Reinke's edema because of some alteration in organization combined with the 5-microm-thick histologic sectioning plane. No significant difference in the degree of elastic system fiber disarrangement was observed between severity grades II and III (p = 0.382). Large areas of disarrangement were predominant (80% of cases)., Conclusions: The disarrangement of elastic fibers in Reinke's edema may cause insufficient tissue resistance and resilience, contributing to the hypermobility observed in Reinke's edema.

Objective: To detect the expression of microRNAs (miRNAs) in laryngeal carcinoma and adjacent normal tissue using microarray, and to discuss the relationship between miRNAs and laryngeal carcinoma., Method: miRNA were extracted from 8 cases of laryngeal cancer tissue and its adjacent normal tissue. miRNA identification were performed by microarray of miRNA hybridization and cluster analysis was used with Significance Analysis of Microarrays (SAM, version 2.1) and Cluster 3.0 software. miRNA were confirmed by real time quantification RT-PCR with RNA-tailing and primer extension., Result: Totally 47 different miRNAs were found expressed in laryngeal cancer, with 23 of miRNA expression were up-regulated and 24 of miRNA expression were down-regulated. The expression of miR-1, miR-486-5p, miR-206, miR-487a,miR-375, miR-422a, miR-144, miR-384, miR-378, miR-133a were down-regulated by 5 multiple and while expression of miR-93, miR-31, miR-20b were up-regulated by 3 multiple. There are 5 miRNA clusters with coexpression in laryngeal cancer tissue and located on chromosome 8, 13, 14, 18 and X. Moreover, RT-PCR analysis demonstrated that there was no significantly difference of miRNA expression between microarray and RT-PCR., Conclusion: The different expression of miRNA may play an important role in the pathogenesis and development of laryngeal cancer.

Any endoscopic diagnostic procedure that is capable of giving exact information on laryngeal lesions without damaging the tissue has essential advantages over standard biopsy. Tissue autofluorescence is defined as a natural ability of tissue to fluoresce when exposed to a certain light wavelength. This feature is a consequence of the presence of fluorophores in the tissues, which are activated by a narrow wavelength range. However, due to their biochemical and biophysical characteristics, laryngeal precancerosis and cancer do not fluoresce when exposed to blue light. In the present study, we used Pentax's System of Autofluorescent Endoscopy (SAFE 1000) to detect autofluorescence disturbances from laryngeal mucosa. Diagnostic parameters (sensitivity and specificity) of the microlaryngoscopy (MLS) and SAFE 1000 in the diagnosis of laryngeal precancerosis and carcinoma were compared and discussed. We have found that SAFE had a better sensitivity with regard to mentioned laryngeal pathology, but MLS had better specificity than SAFE. The overall diagnostic sensitivity in the diagnostics of laryngeal atypical hyperplasia and cancer with SAFE was 89%, as opposed to 73% with MLS. Diagnostic specificity of SAFE for all cases of laryngeal carcinomas and atypical hyperplasia was 78%. The specificity of MLS in diagnostics of laryngeal carcinomas cases was 98%, while that for cases of atypical hyperplasia was 100%. Many other conditions that have impact on autofluorescent features of laryngeal mucosa were also discussed.

Background: Radiation therapy (RT) to the head and neck often results in damage to the vocal folds (VF) and surrounding structures. Characterization and treatment of these sequelae is limited, likely due to the lack of experimental models., Methods: Larynges from rats exposed to 2 fractionation schedules (40 Gy total) were analyzed histologically. In vitro, reactive oxygen species (ROS) synthesis, and transcription of select genes associated with ROS, inflammation, and fibrosis were examined in VF fibroblasts after single-dose radiation., Results: Although radiation-induced histologic alterations are made to VF architecture, 1 fractionation schedule was accompanied by significant morbidity and mortality. In vitro, radiation increased ROS synthesis and inflammatory and profibrotic gene expression., Conclusion: Our data suggest that hyperfractionated RT is more tolerable. Utilizing this model, RT-induced histologic aberrations are made to the VF mucosa. In addition, a relationship between radiation, ROS, and inflammatory and fibrotic gene expression was observed in vitro.

Objectives/hypothesis: Exposure of pig laryngeal mucosa to pepsin and acid will have a differential damaging effect depending on the anatomical site, mirroring the effects seen in the human larynx in laryngopharyngeal reflux (LPR). This study aims to quantitate damage caused to laryngeal tissue by acid alone, and acid and pepsin, and also to determine if the extent of this damage depends on the tissue site., Study Design: Prospective translational research study., Methods: An excised porcine laryngeal damage model in a small Ussing chamber was used to measure the effect of pepsin and acid on five sites (ventricles, vocal folds, posterior commissure, supraglottic, and subglottic mucosa). The tissue samples were incubated on the lumenal side for 1 hour with pH 2 and 4 HCl, pH 2 plus 1 mg/mL pepsin, and pH 4 plus 1 mg/mL pepsin. Damage was assessed by changes in absorbance of the bathing solution at optical density (OD) 260 nm and OD 280 nm and by measurement of released DNA compared to tissues bathed in pH 7.4 buffer. Damage was also assessed histologically., Results: Based on histology, all the tissues were resistant to pH 4.0 except the subglottic mucosa. Only the posterior commissure was not damaged by pH 2.0 plus pepsin. Similar patterns were observed with absorbance changes and DNA release., Conclusions: The subglottic mucosa was the most susceptible to damage and the posterior commissure the least. Laryngeal tissues are essentially resistant to damage at pH 4.0, but are damaged when pepsin is present. This suggests that in LPR, pH 4.0 or above refluxate would only be damaging if it contains pepsin.

Objective: Several classification systems for Reinke's edema have been proposed in the past, which are somewhat less than morphologically ideal. The objective of the present study is to evaluate the ability of optical coherence tomography (OCT) to attain a reproducible graduation of Reinke's edema., Material and Methods: In a prospective study, 30 consecutive patients underwent endolaryngeal, microsurgical resection of their Reinke's edema. Graduation was performed through OCT based on morphologic criteria, where each result was compared with that of other classification systems., Results: In Reinke's edema grade I according to Glanz, a feathered pattern is found, while Reinke's edema grade II demonstrates a lacunar and Reinke's edema grade III a confluent pattern. Correlation between the different classification systems was uniformly weak and not significant., Conclusions: Due to the three different recognizable patterns, OCT qualifies as an objective and reproducible aid for the graduation of Reinke's edema and gives certain insights in its pathogenesis., (Georg Thieme Verlag KG Stuttgart, New York.)

Purpose: To evaluate swine vocal fold healing in a period of 30 days after topical mitomycin-C application., Methods: Twelve swine underwent exeresis of mucosal flaps from the free edge of the anterior third of both vocal folds with a cold instrument (laryngeal scissors). The animals were divided into two groups: EG (Experimental Group), consisting of 6 animals undergoing topical MMC application (4 mg/dL) on the operated area for 4 minutes; CG (Control Group), 6 animals undergoing topical saline solution application on the operated area for 4 minutes. After 30 days, the animals were sacrificed and the larynges were collected and examined for the presence of synechiae as well as a histological immunohistochemical assessment of immature and mature collagen deposition, number of blood vessels and myofibroblasts., Results: Mature collagen deposition in the EG was 452.12 microm(2) and 1332.31 microm(2) in the CG; immature collagen deposition was 1511.73 microm(2) in the EG and 1020.61 microm(2) in the CG. The number of myofibroblasts was 1.556 in the EG and 3.583 in the CG. The number of blood vessels was 2.565 in EG and 6.917 in the CG. There were no synechiae in the two studied groups., Conclusions: There was an increase in immature collagen deposition in the experimental group when compared with the control group. There was a decrease in mature collagen deposition in the experimental group when compared with the control group. There was a decrease in the number of myofibroblasts in the experimental group when compared with the control group. A decrease in blood vessels was observed in the experimental group when compared with the control group. There was no synechia formation in either studied group.

Objectives: Contact endoscopy is a noninvasive tool that allows in vivo and in situ examination of superficial mucosa. Its use for early diagnosis of cancerous lesions of the oropharynx and larynx has not been evaluated. The aim of the study was to validate contact endoscopy for the examination of pharyngeal and laryngeal mucosa., Study Design: Prospective clinical study., Methods: Superficial cells of the mucosa were stained with methylene blue and examined with contact endoscopes. The documented images were assessed by a cytopathologist and by an otolaryngologist independently for each patient. Biopsies for histopathological examination of the area were performed and correlated with contact endoscopic findings of both examiners., Results: Of the 42 examined specimen, 32 (76.2%) showed benign changes in the histological analysis. Squamous cell carcinoma was revealed in 10 specimen (23.8%). Using contact endoscopy, the cytopathologist accurately identified 90.6% of the benign findings (29 of 32); however, only seven of 10 (70%) carcinomas were correctly categorized. In comparison, the otolaryngologist made a correct diagnosis in 93.75% (30 of 32) of the benign and in 90% (nine of 10) of the malignant cases. Thus, a sensitivity of 90% and a specificity of 93.75% can be achieved by contact endoscopy., Conclusions: Contact endoscopy offers valuable support for the evaluation of oropharyngeal, hypopharyngeal, and laryngeal mucosa. Contact endoscopy can be a useful contribution to rapid intraoperative evaluation of mucosal alterations for early diagnosis of tumors and might reduce diagnostic biopsy sampling. Even so, it does not replace biopsy sampling.

Primary candidiasis in the laryngeal mucosa is rare but has many predisposing factors, in particular immune insufficiency and long-term antibiotic use. This study involved a reassessment of 530 laryngeal biopsy specimens originally examined between 2004 and 2009, from which 11 cases of hyperplastic laryngeal candidiasis were identified according to their clinical and histopathological features. Among possible predisposing factors, immune insufficiency was identified in three patients, there was long-term antibiotic use in two cases, use of inhaled corticosteroids in four cases, smoking in eight cases and reflux symptoms in four cases. Dysplasia as well as hyperplasia was seen in two cases. Endolaryngeal lesion excision was performed in eight cases and biopsy leading to radical surgery was done in three. In clinical follow-up no lesions were observed in nine cases while two had new leukoplakic lesions.

Experimental chronic laryngitis (ECL) was induced in 34 adult male rabbits by placement of fosta nylon thread into their trachea. Changes in the laryngeal mucous membrane in ECL were studied at days 30-90 using histological, histochemical and morphometric methods. ECL resulted in the thickening of both the epithelium and lamina propria of the laryngeal mucous membrane. In the ECL dynamics, the thickness of stratified squamous epithelium was increased insignificantly, however, it demonstrated the stimulation of the focal hyperplastic processes. Lamina propria, underlying this epithelium, was found to become thicker throughout the whole experiment (days 30-90). At the same time, progressive growth of the thickness of both pseudostratified ciliated epithelium and the lamina propria beneath was observed. This was accompanied by the hyperplasia of laryngeal endocrine cells and the increase of their secretory activity.

The vocal fold mucosa plays an important role in voice production. Its cellular composition and density frequently change under various pathological conditions, often contributing to altered extracellular matrix production, tissue viscoelasticity, and voice quality. In this study, cellular changes in the rat mucosa following a unilateral stripping injury were investigated and analyzed semi-quantitatively. Distinctive and sequential changes in cellular morphology, composition, and density were observed in the mucosa post-injury. Cellular recruitment was a major event during the early stage of injury and reached its peak level by day 5 post-injury. Several types of cells, including neutrophil-like cells, epithelial cells, and fibroblast-like cells, were sequentially recruited. The sequential emergence of reactive cell populations following injury and subsequent reconstruction of the mucosa suggests their involvement in vocal fold tissue repair and scar formation processes.

Objectives: Epidermal Growth Factor Receptor variant III (EGFRvIII) has been believed to be an attractive tumor-specific candidate for molecular targeting therapy. However, there is little literature dealing with this variant of EGFR expressed in laryngeal carcinomas. In the present study, we try to evaluate the expression of EGFRvIII, as well as EGFR, in laryngeal carcinoma tissues and its correlation with clinicopathological features., Methods: Real-time polymerase chain reaction (real-time PCR) with TaqMan probes was applied to detect the expression for EGFR and EGFRvIII mRNA in the 39 pairs of samples of laryngeal carcinoma tissues and microscopically normal laryngeal mucosal tissues adjacent to the tumor. 2(-DeltaDeltaCT) method was used to obtain the relative quantity of target mRNA expression. The correlation between EGFRvIII expression and its clinicopathological features was analyzed by Pearson's chi-squared test., Results: Among the 39 pairs of samples of laryngeal carcinoma tissues and microscopically normal laryngeal mucosal tissues adjacent to the tumor, the level of EGFR mRNA of the former (0.030+/-0.076) was higher than that of the latter (0.011+/-0.046) (P<0.01). EGFRvIII mRNA was detected only in six samples of laryngeal carcinoma tissues. While, as control, in 39 samples of microscopically normal laryngeal mucosal tissues, EGFRvIII mRNA was hardly detected. As analyzing the correlation between expression of EGFRvIII and EGFR, we found the positive rate of EGFRvIII expression was higher in samples with relative EGFR mRNA value > or =0.025 than those of EGFR mRNA <0.025. The difference between them was statistically significant (P<0.05)., Conclusions: Expression of EGFRvIII in laryngeal carcinoma was confirmed in this study. It is tumor-specific and tends to be more frequent in EGFR-over expressing tumor tissues and poorly differentiated ones, which may in part contribute to the malignant phenotype.

A mucosal bridge of the true vocal fold is a rare, benign anatomical finding that can cause dysphonia. It has been described by some in the literature as "occult" as it is often not visibly evident on flexible nasopharyngolaryngoscopy or strobovideolaryngoscopy, but mistakenly diagnosed as a sulcus vocalis (Sataloff RT, Rosen C, Hawkshaw M. Occult mucosal bridge of the vocal fold. Ear Nose Throat J. 1997; 76(12):850).(2) Final diagnosis is usually not made until microscopic direct laryngoscopy is performed and palpation of the true vocal fold reveals the mucosal bridge (Tanaka S, Hirano M, Umeno H, Tanaka Y. Mucosal bridge of the vocal fold [Japanese].(4)Nippon Jibiinkoka Gakkai Kaiho. 1991; 94(12):1853-1856). We describe a case of a 15-year-old boy complaining of long-standing hoarseness and found to have bilateral mucosal bridges of the true vocal folds. Previous reports cite cases of a unilateral mucosal bridge. We believe this is the first reported case of bilateral mucosal bridges.

The objectives of this study were to evaluate morphologic alterations and precancerous lesions in Reinke's edema. Patients included were 54 smokers with bilateral Reinke's edema submitted to surgery in the Otolaryngology Department, Botucatu Medical School, São Paulo State University, Brazil, between 2002 and 2006. All specimens were evaluated by light microscopy and five contralateral lesions were also evaluated by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The main histological alterations were edema (100%), inflammation (81.48%), basal membrane (bm) thickening (77.77%), and vessel proliferation (75.92%). Epithelium alterations were classified as grade 0 (11.11%), grade 1 (70.37%), grade 2 (14.81%), and grade 3 (3.70%). In the case included in grade 3 classification, microinvasive carcinoma was detected. SEM showed epithelial surface with some cellular desquamation, few microridges, and a polished and impermeable surface aspect. TEM showed epithelial hyperplasia, basal cells with different sizes, widening of the intercellular spaces, abnormal desmosome architecture, thickening of the bm, some electron-dense vesicles, and points of interruption. The morphological alterations presented in this study are not specific to Reinke's edema but this lesion can be the site of different grades of dysplasia and carcinoma, justifying the importance of periodic laryngeal endoscopic exams and meticulous histological analysis.

The assessment of gene expression profile in laryngeal cancer allows implementation of molecular biology methods in diagnostics, as well as in prognosticating the course of disease, thus allowing taking most optimal decisions as regards the method of treatment, scope of surgical procedure, or the necessity of adding complementary radiotherapy. The aim of the project was to analyze the gene expression profile in laryngeal cancer using oligonucleotide microarrays, having in mind searching new molecular markers for that carcinoma. The study comprised a group of 43 patients (38 males and 5 females) suffering from squamous cell laryngeal carcinoma, diagnosed and surgically treated in the years 2005-2007 in the ENT Department of the Silesian Medical University in Katowice, Poland. RNA was isolated from frozen tissue fragments, with the use of columns RNeasy Midi and Mini Kit (Qiagen). For the examination of gene expression profile, oligonucleotide microarrays of high density were used, provided by Affymetrix (U 133 2.0 PLUS) containing over 54,000 probes for over 47,000 transcripts. Four genes previously not examined in that respect in laryngeal carcinoma, occurred to be good markers of the neoplasm. They are: metal-proteinase ADAM12, cyclin-dependent kinase 2-CDK2, kinesin 14-KIF14, suppressor 1 of checkpoint-CHES1. The analysis of gene expression profile allows, in laryngeal carcinoma, to point out to new genes, which in future may become molecular markers of the carcinoma.

Objective/hypothesis: Oxidative damage mediates the lower airway response to cigarette smoke (CS). In the vocal folds, the tissue phenotype is both distinct and largely uncharacterized. We sought to investigate the effects of CS on the oxidative status and fibroblast phenotype in the vocal folds. Specifically, we hypothesized that CS would induce a hypermetabolic fibroblast phenotype and altered oxidative metabolism potentially providing insight into the relationship among CS, Reinke's edema (RE), and malignancy., Study Design: In vivo/in vitro., Methods: Heme oxygenase (HO)-1 gene expression was examined in human tissue. In vitro, the effects of cigarette smoke condensate (CSC) on HO-1 gene expression and secretion was assayed. In addition, CS-mediated intracellular reactive oxygen species synthesis was quantified, and compared to the response in pulmonary fibroblasts (HFL). We then examined the effects of CSC on migration and proliferation in human vocal fold fibroblasts (HVOX)., Results: : HO-1 expression was approximately 4-fold higher in RE samples versus vocal fold polyps. CSC induced HO-1 gene expression and secretion in a time- and dose-dependent fashion in vitro. CSC also increased intracellular ROS in both HVOX and HFL. CSC decreased HVOX proliferation and migration in a dose-dependent manner., Conclusions: These data suggest that the fibroblast phenotype is influenced by smoke. Our data suggest that the antioxidant response in the vocal fold tissue may be one mechanism of chemoprotection, a putative explanation for the observation that RE rarely transforms to malignancy. In addition, CSC does not appear to induce a hypermetabolic fibroblast phenotype as expected.

Objective: To determine the significance of laryngopharyngeal reflux (LPR) in benign vocal mucosal lesions., Study Design and Setting: A case-control study at the tertiary referral medical center., Subjects and Methods: From April 2003 to December 2006, we studied 110 patients with benign vocal mucosal lesions who had undergone 24-hour ambulatory double pH monitoring. The control group included 200 patients who had undergone ambulatory 24-hour double-probe pH monitoring due to laryngopharyngeal reflux-related symptoms without specific findings of benign vocal mucosal lesions. Reflux symptom index and reflux finding score were measured. We compared the prevalence of pathologic laryngopharyngeal reflux and various parameters of the pH monitoring such as total reflux number, fraction time of pH below 4 in various positions, and DeMeester scores., Results: The prevalence of pathologic laryngopharyngeal reflux was 65 percent in the control group, 66 percent in vocal nodule group, 75 percent in the vocal polyp group, and 90 percent in the Reinke's edema group. Patients with Reinke's edema had a significantly higher prevalence of pathologic laryngopharyngeal reflux than controls (P = 0.016). LPR was associated with a significantly increased risk of Reinke's edema (odds ratio: 4.846, 95% confidence interval 1.093 approximately 21.492). Total reflux number and DeMeester scores in the Reinke's edema group and fraction time of pH below 4 in the supine position in the vocal polyp group were significantly higher than those in the control group., Conclusion: Laryngopharyngeal reflux might play a role as an etiologic factor in Reinke's edema and vocal polyps.

Objectives: The management of vocal fold scarring and sulcus vocalis is challenging. These disorders are thought to be fibroplastic anomalies in the cover portion of the vocal fold that cause deterioration of the vibratory properties of the vocal fold mucosa. Histologic studies have revealed disorganization of extracellular matrix that needs to be addressed in the treatment of scarred vocal folds. Replacement of scar tissues with an appropriate implant may lead to regeneration of the vocal fold mucosa and its tissue properties. This retrospective case study examined the feasibility of using an atelocollagen sheet as a regenerative implant., Methods: Six patients with a post-cordectomy scar or sulcus vocalis underwent implantation of an atelocollagen sheet into the lamina propria of the vocal folds. The procedure consisted of elevation of a microflap, dissection and removal of scar tissue, implantation of the material, and wound closure. Vocal function was evaluated before and after surgery by stroboscopic examination and by aerodynamic and acoustic analyses., Results: The postoperative changes of aerodynamic and acoustic parameters varied among patients; however, gradual improvement was seen in most cases over a year. Stroboscopic findings also revealed gradual improvement of vibratory properties in most cases., Conclusions: Implantation of an atelocollagen sheet may have restorative effects on vocal fold scarring and sulcus vocalis in terms of tissue properties and function of the mucosa.

Objective: To investigate the expression of EMS1 in laryngeal carcinoma and its clinical significance., Method: The expression of EMS1 protein was measured in 40 samples of, 40 samples of para carcinoma tissues (which were near to cutting margin of laryngeal carcinoma tissue over 0.5 cm), and 20 samples of normal laryngeal mucosa as controls by Flow Cytometer (Epics-XL II)., Result: The quantity and percentage of EMS1 protein expression in laryngeal carcinoma tissues was significantly higher than those in para carcinoma and in normal laryngeal mucosa tissues respectively (P<0.05). There was no significant expression difference between the para carcinoma tissues and normal laryngeal mucosa tissues. There were positive correlation between the expressions of EMS1 protein and metastasis, pathological grade and clinical stage in laryngeal carcinoma. But there were not relationship with patients' clinical classification, tumor size, smoking history, age and sex., Conclusion: The high expression of EMS1 may contribute to the carcinogenesis and development of laryngeal carcinoma. The expression of EMS1 protein is an important index of judging differentiation, infiltration, metastasis and staging of laryngeal carcinoma.

Reinke's space is a highly specific structure critical for the function of the vocal fold, involved in a majority of pathological changes of the larynx. The aim of the study was to contribute to the understanding of edematous areas of vocal fold mucosa in patients with Reinke's edema using contact telescopy (ConTel). The edematous tissue which could be identified grossly by microlaryngoscopy under general anesthesia was stained topically with methylene blue and then examined using ConTel. The observed changes on contact images were further correlated with corresponding histological sections of biopsied edematous tissue. In patients with Reinke's edema examined using ConTel, we observed longitudinal arranged mucosal "blue lines" (BL). In the histological sections of edematous tissue showing the BL on ConTel, we almost regularly observed well-developed hollow spaces in subepithelia. Also, they were regularly oriented from the middle of the membranous portion of the vocal fold toward the arcuate line distributed almost parallel to the free edge of the vocal fold. BL were a specific feature of Reinke's edema as they were not found in control groups with normal vocal fold mucosa and with vocal fold polyps. For the first time we are describing a new superficial morphological feature of Reinke's edema. The BL could present epithelial ConTel markers of the area between two enlarged neighboring subepithelial crevices of Reinke's space, as indicated on histological sections. Further characterization of BL is required for the understanding of their role in Reinke's edema.

Objective: To study the clinical effect of heterogeneity (cattle) acellular dunal matrix in repairing mucosa defect in laryngeal surgery., Methods: Eighteen cancer patients with mucosa defect in central vocal area accepted treatment with heterogeneity acellular dunal matrix after surgery. There were two methods to repair mucosa defect. One was simple use of acellular dunal matrix, the second was combined use of acellular dunal matrix and muscle lamella or muscle and tendon film lamella. 18 cases had cancer in central vocal area: T2N0M0 (8), T3N1M0 (5), T3N2M0 (4), T4N2M0 (1). All were squamous cell carcinoma. Ten cancer patients accepted radiation after surgery. The radiotherapy volume was 60-80 Gy. After the operation, the patients were checked by fibrolaryngoscope four or five times after half a year, observing the dynamic development., Results: All 18 patients were healed, rechecked by endoscope after 0.5-6 months, heterogeneity acellular dunal matrix mingled with mucosa within 30-60 d, no allergy and irritation were found. The laryngeal function, including breathing, pronouncing and swallowing, was recovered. The survival rate (1 year) was 100%, and 10 patients survived after 2 years. After radiotherapy, the process of recovery was not affected., Conclusions: Heterogeneity acellular dunal matrix can be easily obtained and it is a new method to repair mucosa defect. The operative procedure is easy to perform and worthwhile to use clinically.

Objective: To investigate the expression and clinical significance of oncogene AKT2 in laryngeal squamous cell carcinoma (LSCC)., Method: The expression of AKT2 protein was measured with immunohistochemistry (Elivision two steps) in 60 cases of LSCC tissue, 28 cases of peri-cancer tissue and 16 cases of normal laryngeal epithelium, the relationship between AKT2 expression and clinical pathologic factors in LSCC was also analyzed., Result: The positive rates of AKT2 protein expression in LSCC and peri-cancer tissue were 56.7% (34/60), 14.3% (4/28) respectively. There was no expression of AKT2 in normal laryngeal epithelium. The positive rate of AKT2 protein expression in LSCC was significantly higher than that in peri-cancer tissue and normal laryngeal epithelium (P < 0.05, respectively). The positive rate of AKT2 was correlated with tumor site, lymph node metastasis and clinical stage of LSCC (P < 0.05, respectively), but wasn't correlated with patient's age, sex, and T stage (P > 0.05, respectively)., Conclusion: AKT2 might play an important role in the tumorigenesis and development of LSCC, and it was suggested that AKT2 could be a useful marker for predicting tumorigenesis, invasion and metastasis potential of LSCC.