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BackgroundNivolumab paved a new way in the treatment of patients with recurrent or metastatic (RM) head and neck squamous cell carcinoma (RM-HNSCC). However, the limited rates of long-term survivors (< 20%) demand a robust prognostic biomarker. This nationwide multi-centric prospective study aimed to identify a plasma exosome (PEX) mRNA signature, which serves as a companion diagnostic of nivolumab and provides a biological clue to develop effective therapies for a majority of non-survivors.MethodsPre-treatment plasmas (N = 104) of RM-HNSCC patients were subjected to comprehensive PEX mRNA analyses for prognostic marker discovery and validation. In parallel, paired treatment-naïve tumor and plasma samples (N = 20) were assayed to elucidate biological implications of the PEX mRNA signature.ResultsAssays for pre-treatment blood samples (N = 104) demonstrated that a combination of 6 candidate PEX mRNAs plus neutrophil-to-lymphocyte ratio precisely distinguished non-survivors from >2-year survivors (2-year OS; 0% vs 57.7%; P = 0.000124) with a high hazard ratio of 2.878 (95% CI 1.639-5.055; P = 0.0002348). Parallel biological assays demonstrated that in the paired treatment-naïve HNSCC tumor and plasma samples (N = 20), PEX HLA-E mRNA (a non-survivor-predicting marker) was positively corelated with overexpression of HLA-E protein (P = 0.0191) and the dense population of tumor-infiltrating NK cells (P = 0.024) in the corresponding tumor, suggesting that the HLA-E-NKG2A immune checkpoint may inhibit the antitumor effect of PD-1blockade.ConclusionThe PEX mRNA signature could be useful as a companion diagnostic of nivolumab. The combination of an anti-NKG2A antibody (i.e., monalizumab) and nivolumab may serve as a treatment option for non-survivors predicted by a RT-qPCR-based pre-treatment measurement of PEX mRNAs.

BackgroundIn view of improving biomarkers predicting the efficacy of immunotherapy for head and neck squamous cell carcinoma (R/M HNSCC), this multicenter retrospective study aimed to identify clinical, tumor microenvironmental, and genomic factors that are related to therapeutic response to the anti- Programmed cell death protein 1 (PD-1) antibody, nivolumab, in patients with R/M HNSCC. MethodsThe study compared 53 responders and 47 non-responders, analyzing formalin-fixed paraffin-embedded samples using 14-marker multiplex immunohistochemistry and targeted gene sequencing.ResultsOf 100 patients included, responders had significantly lower smoking and alcohol index, higher incidence of immune related adverse events, and higher PD-1 ligand (PD-L1) expression in immune cells as well as PD-L1 combined positive score (CPS) than non-responders. The frequency of natural killer cells was associated with nivolumab response in patients with prior cetuximab use, but not in cetuximab-naïve status. Age-stratified analysis showed nivolumab response was linked to high CPS and lymphoid-inflamed profiles in patients aged ≥ 65. In contrast, lower NLR in peripheral blood counts was associated with response in patients aged < 65. Notably, TP53 mutation-positive group had lower CPS and T cell densities, suggesting an immune-excluded microenvironment. Patients with altered tumor suppressor gene pathways, including TP53, CDKN2A, and SMAD4 mutations, had lower CPS, higher smoking index, and were associated with poor responses. ConclusionNivolumab treatment efficacy in HNSCC is influenced by a combination of clinical factors, age, prior treatment, immune environmental characteristics, and gene mutation profiles.

Abstract Objectives The growth and development of the epiglottis and preepiglottic space (PES) of the human larynx as it acquires the vocal tract were investigated. Methods Three newborns, one infant, four children (2, 7, 8, and 12 years old), and two adult normal larynges were investigated and compared using the whole organ serial section technique. Results The newborn PES occupied a small area just anterior to the epiglottis. It was composed of immature adipose tissue and areolar tissue. The epiglottis lay on a somewhat horizontal axis and is partially obscured behind the hyoid bone. The hyoid bone overlapped the thyroid cartilage, partially obscuring the superior thyroid notch. The newborn epiglottic cartilage was immature elastic cartilage, and the elastic fiber component was sparse. In the first 8 years of life, as the PES grew, the PES was located not only anterior to but also posterolateral and inferolateral to the epiglottic cartilage and thyroepiglottic ligament. Meanwhile, the epiglottic cartilage matured. Conclusions In order to develop the vocal tract for speech production, it is reported that the human larynx descends as the child grows in the first 9 years of life. This study showed that the PES, occupying a small area just anterior to the epiglottis, grew and existed astride the epiglottis as the larynx descended and the vocal tract developed. Consequently, its distribution allows the epiglottis to more effectively play the role of retroflection during swallowing in order to prevent aspiration. The human speech faculty likely develops in conjunction with swallowing physiology.

Abstract Objective Pretreatment systemic inflammation and nutrition‐based prognostic indices (SINBPI) have demonstrated significance. This study investigated the prognostic value of pretreatment SINBPI for patients with oropharyngeal cancer and identified unfavorable prognostic markers. Methods We retrospectively reviewed the data of 124 patients with oropharyngeal squamous cell carcinoma (OPSCC) who received definitive treatment between January 2010 and December 2018. The prognostic utility of the neutrophil‐to‐lymphocyte ratio, platelet‐to‐lymphocyte ratio (PLR), lymphocyte‐to‐monocyte ratio (LMR), prognostic nutritional index, and high‐sensitivity modified Glasgow prognostic score (HS‐mGPS) was assessed for disease‐free survival (DFS), disease‐specific survival (DSS), and overall survival (OS) using univariate and multivariate analyses. Results Multivariate analyses revealed that human papillomavirus (HPV) status and HS‐mGPS were significantly associated with DFS, DSS, and OS. Patients with a HS‐mGPS of 2 had a significantly higher rate of treatment‐related deaths than those with a HS‐mGPS of 0 or 1. The combination of the HS‐mGPS and PLR had more accurate predictive ability in DFS and OS compared with the HS‐mGPS alone, and the combination of the HS‐mGPS and LMR had more accurate predictive ability in DSS and OS. Conclusion Our results indicated that the HS‐mGPS was a useful prognostic marker for patients with OPSCC, and combined markers consisting of the HS‐mGPS and PLR or LMR may provide more accurate prognostic predictions. Level of Evidence: 3.

Abstract Objectives To analyze various aspects of complex tissue, there is increasing demand to study each sample at different length scales in biology. Correlative light and electron microscopy (CLEM) is the latest technique to correlate two different types of information on the exact same histological area of interest: histology (from light microscopy) and ultrastructure (from electron microscopy). The three‐dimensional fine structures of the maculae flavae (MFe) of the human vocal fold were investigated using CLEM. Methods Five normal human adult vocal folds as specimens embedded in paraffin, sectioned, and mounted on glass slides with/without a chemical digestion method (modified sodium hydroxide maceration method) were investigated. Observations using CLEM were performed. Results The fine structures of cells and extracellular matrices in the MFe and their peripheral regions were able to be observed on the exact same histological area of interest with the light microscope and field emission‐scanning electron microscope. Cobblestone‐like polygonal cells, vocal fold stellate cell‐like cells, and fibroblast‐like spindle cells were intermingled in the MFe of the human vocal fold. The extracellular matrices surrounding each three types of cell in the MFe differed, suggesting the cells were different in functional property. Conclusion CLEM is a useful technique to observe the three‐dimensional fine structures of the human vocal fold mucosa. The results of the present study are consistent with the hypothesis that the cells in the MFe of the human vocal fold have heterogeneity and each three types of cell have different properties.

IntroductionExpression of the NTRK gene is rare in solid tumors but is highly prevalent in salivary gland secretory carcinomas. Here, we report a case of a complete response to entrectinib in a patient with NTRK fusion gene-positive parotid carcinoma.Case descriptionThe patient was a 44-year-old man who underwent total left parotidectomy and left cervical lymph node dissection for a left parotid tumor at 24 years of age. The histopathological diagnosis was mammary analog secretory carcinoma. Postoperatively, the patient received only radiation therapy. Sixteen years after the surgery, the patient became aware of a mass in the left parotid region. A close examination revealed local recurrence and multiple cervical lymph node metastases. S-1 monotherapy was started as chemotherapy but was discontinued 3 years later because of disease progression. As there was no standard treatment, a comprehensive genomic profiling test using a next-generation sequencer was performed, and the ETV6-NTRK3 fusion gene was identified. Entrectinib, an NTRK inhibitor, was immediately administered at a dose of 600 mg/day. The local recurrence rapidly shrank grossly from the beginning of treatment, and a complete response was observed 6 months later. However, creatinine levels exhibited an increase at week 68 of treatment; consequently, entrectinib dosage was lowered to 400 mg/day, leading to an immediate improvement in creatinine levels. Entrectinib was associated with additional side effects, including dysgeusia, fatigue, dizziness, and weight gain, all of which were also alleviated by the reduction in entrectinib dose. Thirty months after treatment initiation, the patient maintained a complete response and continued to receive entrectinib.ConclusionThe NTRK fusion gene should always be checked in the presence of salivary gland secretory carcinoma.

Abstract Head and neck cancers, especially in hypopharynx and oropharynx, are often detected at advanced stage with poor prognosis. Narrow band imaging enables detection of superficial cancers and transoral surgery is performed with curative intent. However, pathological evaluation and real‐world safety and clinical outcomes have not been clearly understood. The aim of this nationwide multicenter study was to investigate the safety and efficacy of transoral surgery for superficial head and neck cancer. We collected the patients with superficial head and neck squamous cell carcinoma who were treated by transoral surgery from 27 hospitals in Japan. Central pathology review was undertaken on all of the resected specimens. The primary objective was effectiveness of transoral surgery, and the secondary objective was safety including incidence and severity of adverse events. Among the 568 patients, a total of 662 lesions were primarily treated by 575 sessions of transoral surgery. The median tumor diameter was 12 mm (range 1–75) endoscopically. Among the lesions, 57.4% were diagnosed as squamous cell carcinoma in situ. The median procedure time was 48 minutes (range 2–357). Adverse events occurred in 12.7%. Life‐threatening complications occurred in 0.5%, but there were no treatment‐related deaths. During a median follow‐up period of 46.1 months (range 1–113), the 3‐year overall survival rate, relapse‐free survival rate, cause‐specific survival rate, and larynx‐preservation survival rate were 88.1%, 84.4%, 99.6%, and 87.5%, respectively. Transoral surgery for superficial head and neck cancer offers effective minimally invasive treatment. Clinical trials registry number: UMIN000008276.

Abstract Objectives Our previous investigations showed that tissue stem cells in the maculae flavae (a stem cell niche) form colonies in vivo like stem cells in vitro. However, the roles of colony‐forming cells in the maculae flavae in vivo have not yet been determined. This study investigated the metabolism of the colony‐forming tissue stem cells in the maculae flavae of the human adult vocal fold. Study design Histologic analysis of the human vocal folds. Methods Three normal human adult vocal folds were investigated under transmission electron microscopy and light microscopy including immunohistochemistry. Results Mitochondrial cristae of the colony‐forming cells in the maculae flavae were sparse. Hence, the microstructural features of the mitochondria suggested that their metabolic activity and oxidative phosphorylation were low. Colony‐forming cells strongly expressed glucose transporter‐1 and glycolytic enzymes (hexokinase II, glyceraldehyde‐3‐phosphate dehydrogenase and lactate dehydrogenase A). The colony‐forming cells did not express phosphofructokinase‐1 but did express glucose‐6‐phosphate dehydrogenase indicating the cells relied more on the pentose phosphate pathway. Since the colony‐forming cells expressed lactate dehydrogenase A, cells seemed to rely more on anaerobic glycolysis in an anaerobic microenvironment. Conclusions The present study is consistent with the hypothesis that the colony‐forming tissue stem cells in the maculae flavae of the human adult vocal fold seemed to rely more on anaerobic glycolysis using the pentose phosphate pathway for energy supply in vivo. Microstructural features of the mitochondria and expressed glycolytic enzymes of the colony‐forming cells in the maculae flavae suggested that the oxidative phosphorylation activity was low. In an anaerobic microenvironment in vivo, there is likely a complex cross‐talk regarding the metabolism between the colony‐forming aggregated cells along the adhesion machinery and chemical signaling pathways, which reduces toxic oxygen species and is favorable to maintaining the stemness and undifferentiated states of the tissue stem cells. Level of Evidence NA.

Abstract Objectives In our previous studies, the features of the mitochondria of tissue stem cells in the maculae flavae of the human vocal fold suggested that their metabolic activity and oxidative phosphorylation was low. This study investigated the metabolic activity, especially glycolysis of the tissue stem cells in the maculae flavae of the human adult vocal fold. Study Design Histologic analysis of the human vocal folds. Methods Three normal human adult vocal folds obtained from autopsy cases were investigated using immunohistochemistry. Results Among the three phenotypes of cells in the human adult maculae flavae, the vocal fold stellate cell‐like cells strongly expressed glucose transporter‐1. Three phenotypes of cells in the human adult maculae flavae expressed glycolytic enzymes (hexokinase II, glyceraldehyde‐3‐phosphate dehydrogenase and lactate dehydrogenase A) indicating the tissue stem cells in the maculae flavae relied more on glycolysis. The cells did not express phosphofructokinase‐1 but did express glucose‐6‐phosphate dehydrogenase indicating the cells relied more on the pentose phosphate pathway. The cells expressed lactate dehydrogenase A indicating the maculae flavae of the human adult vocal fold was likely to be an anaerobic microenvironment. Conclusions The present study is consistent with the hypothesis that the tissue stem cells in the maculae flavae of the human vocal fold seem to rely more on anaerobic glycolysis, especially by the pentose phosphate pathway, for energy supply. The metabolism of the tissue stem cells in the maculae flavae of the human adult vocal fold is likely to prevent toxic reaction oxygen species and be favorable to maintaining the stemness and undifferentiated states of the tissue stem cells in the stem cells system. Level of Evidence NA.

Abstract Objectives There is growing evidence that the cells in the maculae flavae (MFe) are tissue stem cells and the MFe are a stem cell niche of the human vocal fold mucosa. Heterogeneity and hierarchy of tissue stem cells in the MFe of newborn vocal fold were investigated in vivo. Study design Histologic analysis of the human vocal folds. Methods Five normal human newborn vocal folds were investigated under transmission electron microscopy and light microscopy. Results Cobblestone‐like polygonal cells, vocal fold stellate cell‐like cells, and fibroblast‐like spindle cells were intermingled in the newborn MFe in vivo, indicating that the cells in the MFe had heterogeneity. However, cobblestone‐like polygonal cells were predominant. Free ribosomes were well developed in the cytoplasm. The cells in some cases formed gap junctions with each other. The cells in some cases were attached to other cells and formed cell junctions with each other. These findings indicated cells in the newborn maculae flavae possessed features of mesenchymal cells (cells in mesenchyme). Colony‐forming‐unit‐like cell aggregate was observed, indicating the cells in the newborn MFe had stemness. The cobblestone‐like polygonal cells expressed SSEA‐3 (a human pluripotent stem cell marker), indicating they were at the top of a cellular hierarchy in the stem cell system. Conclusions The cells in the MFe of the human newborn vocal fold mucosa had heterogeneity and hierarchy in the stem cell system in vivo. At birth, newborn maculae flavae are ready to start the growth of the vocal fold mucosa as a vibrating tissue.

Abstract Objective Histopathology of the maxillary sinus mucosa with intractable odontogenic maxillary sinusitis (OMS) was investigated and the role endoscopic sinus surgery (ESS) plays in its pathophysiology was clarified. Study Design Histopathological analysis of the OMS mucosa. Methods Surgical specimens were obtained from 20 patients who underwent ESS for intractable OMS. For rigid endoscopic observation of the mucosae, a 70° rigid endoscope 4 mm in diameter with an attached high definition surgical camera was used. Histopathological analyses of the maxillary sinus mucosa were conducted by light and scanning electron microscopy. Results All the maxillary sinuses were filled, not with viscous, but with purulent secretions. The high‐definition camera showed that the maxillary sinus mucosa had gyrus‐like appearance. Light microscopic histopathological studies revealed that the surface of the maxillary sinus mucosa was convoluted. Light and scanning electron microscopic histopathological studies revealed that the ciliated cells of the epithelium had not decreased and their goblet cells were not hypertrophic, indicating that the damage of the ciliated columnar epithelium was not severe and they were not injured irreversibly. Conclusion The ciliated columnar epithelium with intractable OMS was not severely damaged and not irreversibly injured. Hence, the pathophysiology of intractable OMS is one of the reasons why ESS is highly indicated for maxillary sinusitis requiring surgery and the treatment results are exceptionally good when the ventilation and drainage of the maxillary sinus is successfully restored after surgery. Level of Evidence NA

Abstract Background This pilot study evaluated the long‐term outcomes of patients with advanced T2 or T3 squamous cell carcinoma of the larynx (SCC‐L) who were treated with selective intra‐arterial cisplatin and concomitant radiotherapy (RADPLAT). Methods We retrospectively investigated the data of 49 patients with advanced T2 or T3 SCC‐L who received a RADPLAT regimen with low‐dose cisplatin. Results The 5‐year locoregional control, disease‐specific survival, and overall survival rates were 83.3%, 88.1%, and 82.6%, respectively, while the 5‐year freedom from laryngectomy, laryngectomy‐free survival, and laryngo‐esophageal dysfunction‐free survival rates were 89.6%, 79.4%, and 77.1%, respectively. The incidences of grade 3‐4 hematologic and nonhematologic toxicities were 18% and 6%, respectively. Although two patients (4%) developed late toxicities within 5 years following RADPLAT, no other events were noted beyond 5 years. Conclusion This pilot study demonstrated that RADPLAT is feasible and safe and yielded favorable survival outcomes and functional laryngeal preservation in patients with advanced T2 or T3 SCC‐L. Level of evidence 3

Abstract Aqueous lithium‐ion batteries are promising electrochemical energy storage devices owing to their sustainable nature, low cost, high level of safety, and environmental benignity. The recent development of a high‐salt‐concentration strategy for aqueous electrolytes, which significantly expands their electrochemical potential window, has created attractive opportunities to explore high‐performance electrode materials for aqueous lithium‐ion batteries. This study evaluates the compatibility of large‐capacity oxygen‐redox cathodes with hydrate‐melt electrolytes. Using conventional oxygen‐redox cathode materials (Li2RuO3, Li1.2Ni0.13Co0.13Mn0.54O2, and Li1.2Ni0.2Mn0.6O2), it is determined that avoiding the use of transition metals with high catalytic activity for the oxygen evolution reaction is the key to ensuring the stable progress of the oxygen redox reaction in concentrated aqueous electrolytes.

Olfactory neuroblastoma is a rare neuroendocrine tumor derived from the olfactory epithelium. We report a case that showed peculiar histology in a late lymph node metastasis. The patient was a 78-year-old Japanese male, who had headache, epistaxis, and nasal obstruction following bloody sputum that was untreated for ten years. The initial biopsy was interpreted as anaplastic carcinoma, and after being referred to a regional university hospital, a total of 60 Gy of radiation therapy was performed prior to the excision of the residual tumor. Neck metastasis was found on the left side two years later, and the patient died of other disease with no recurrence five years after successful neck dissection.Retrospective examination of the biopsy specimen revealed a neurofibrillary matrix besides nesting growth of basophilic naked cells. The neck metastases exhibited similar histology, and a large number of rosette structures containing mucus and extensive squamous metaplasia were observed. Immunohistochemically, naked cells and neurofibrillary matrix were positive for synaptophysin and chromogranin A, and rosette structures and some naked cells were positive for AE1/AE3 and CAM5.2. While epithelial differentiation of olfactory neuroblastoma is a unique feature absent in neuroblastomas in other regions, it was clearly and prominently manifested in the present case.

Herein, we summarize two rare cases of salivary neuroendocrine carcinoma arising in the parotid gland. The patients were a 54-year-old Japanese female and a 74-year-old Japanese male. In the first case, the tumor presented as a typical small cell carcinoma with extensive crush artifacts and nuclear molding, but metastatic foci of the lymph node revealed characteristic ribbon-like trabecular arrangements or sheet-like solid growths of tumor cells with moderate amounts of pale eosinophilic cytoplasm intermingled with bizarre giant or multinucleated cells. In the second case, the tumor was initially diagnosed as a small cell neuroendocrine carcinoma, but a retrospective review revealed that the large cell component was dominant and that it should be strictly considered a mixed large and small cell neuroendocrine carcinoma. The difference in prognoses between these two salivary subtypes remains unknown, and distinction between the small and large cell subtypes of neuroendocrine carcinomas seems merely simple and convenient in the salivary glands.

Abstract Objective High‐resolution manometry (HRM) is used to analyze upper esophageal sphincter (UES) physiology. Conventional HRM can yield imprecise measurements of UES resting pressure given its unidirectional sensors and averaging of circumferential pressure. In contrast, three‐dimensional (3D) measurements can yield precise UES resting pressure data over the entire length of the UES. This study conducted a detailed analysis of UES resting pressure as evaluated by 3D‐HRM. Methods Seventeen young, healthy adult participants (7 females and 10 males) were recruited. The 3D‐HRM system used includes a pressure sensor catheter (outer diameter, 4 mm) and eight‐channel transducers arranged circumferentially to acquire pressure data at 45° intervals. The catheter was inserted transnasally into the esophagus and automatically retracted at a speed of 1 mm/s. Participants performed the following tasks: maintain resting breathing, perform breath holding, and perform the Valsalva maneuver. Data were obtained and compared per millimeter over the longitudinal UES length. Results Clear 3D waveforms were identified, with greater mean UES pressures in anterior‐posterior directions than in lateral directions (P

Objectives There is growing evidence to suggest that the cells in the maculae flavae of the vocal fold mucosa are tissue stem cells of the human vocal fold. This study investigated the metabolic activity of the cells in the maculae flavae of the human vocal fold from the aspect of mitochondrial microstructure. Study Design Histologic analysis of the human vocal folds. Methods Five normal human adult vocal folds obtained from autopsy cases were investigated under transmission electron microscopy. Results Mitochondria were randomly distributed in the cytoplasm of the cells. The morphological features of the mitochondria consisted of a double‐membrane‐bounded body containing matrices and a system of cristae. In each mitochondria, the lamellar cristae were sparse. The intercristal space was occupied by a mitochondrial matrix which contained electron‐dense matrix granules, mitochondrial DNA, and ribonucleoprotein granules. Some mitochondria spread out over or fused to the surface of a lipid droplet in the cytoplasm. In addition, both the mitochondrial outer and inner membranes and the membranes of the lipid droplets had disappeared. Some close association between mitochondria and rough endoplasmic reticulum was present. Conclusions The features of the mitochondria of the cells in the maculae flavae of the human vocal fold suggested that their metabolic activity and oxidative phosphorylation were low and that they may have shifted to the utilization of lipids to some extent for their metabolic needs. Level of Evidence NA

We describe a rare case of epithelioid angiosarcoma (AS) arising in the parotid area and discuss its immunohistochemical features and cytological correlation. The patient was a 55-year-old Japanese male, who was aware of increasing swelling in the left parotid to lateral cervical area for 1 month. Clinical diagnosis as a parotid gland cancer was made on basis of imaging study followed by fine needle aspiration cytology, which was interpreted as a high-grade adenocarcinoma. He underwent a radical surgical resection, but died of the residual tumor rapidly growing associated with distant metastases 3 months after surgery. Histologically, the tumor comprised round to ovoid or polygonal tumor cells exhibiting marked pleomorphism and a predominant sheet-like arrangement along with extensive soft tissue invasion. Tumor cells were immunoreactive for CD31 and factor VIII, but negative for CD34. In cytological specimens, tumor cells appeared as isolated single cells and in loosely attached groups or three-dimensional cohesive clusters in the necrotic background. The cells varied in size and shape, and had remarkably enlarged and eccentrically located nuclei with prominent nucleoli. Intracytoplasmic lumen was occasionally recognized. Based on the microscopic and immunohistochemical findings, the present tumor was diagnosed as an epithelioid AS. Epithelioid AS may occasionally arise in the maxillofacial areas and be confused with undifferentiated carcinoma or adenocarcinoma because of epithelioid cytology and unique immunohistochemical profiles such as possible expression of cytokeratins and a lack of CD34. Epithelioid AS should be included in differential diagnoses of high-grade epithelioid malignancies in this region.

We retrospectively analyzed 14 patients with locally advanced squamous cell carcinoma of ethmoid sinus (LASCC-ES) for the feasibility of anterior craniofacial resection (ACFR). Ethmoid cancer treatment comprised alternating chemoradiotherapy (ALCRT; n = 1), concomitant radiotherapy and intra-arterial cisplatin (RADPLAT; n = 4) and ACFR (n = 9). The 3- and 5-year overall survival (OS) rates of patients were 47.6 and 39.6%, respectively. The 3-year local control (LC) rates of chemoradiotherapy (CRT; ALCRT and RADPLAT) (n = 5) and ACFR (n = 9) groups were 0 and 66.7% (p = 0.012), respectively. The 3-year progression-free survival (PFS) rate of the CRT and ACFR groups were 0 and 55.6% (p = 0.018), respectively. The 3-year OS rate of the CRT and ACFR groups were 0 and 76.2% (p = 0.005), respectively. Postoperative pathological examinations confirmed positive margins in 3 (33%) of 9 cases. The 3-year LC and PFS rates of cases (n = 3) with positive surgical margins were significantly poorer than those of cases (n = 6) with negative surgical margins. Although ACFR for LASCC-ES is a feasible treatment, cases with positive surgical margins were more prone to local relapse. Therefore, surgical safety margins should be thoroughly assessed.

OBJECTIVES:Scarred vocal folds result in irregular vibrations during phonation due to stiffness of the vocal fold mucosa. To date, a completely satisfactory corrective procedure has yet to be achieved. We hypothesize that a potential treatment option for this disease is to replace scarred vocal folds with organotypic mucosa. The purpose of this study is to regenerate vocal fold mucosa using a tissue-engineered structure with autologous oral mucosal cells. STUDY DESIGN:Animal experiment using eight beagles (including three controls). METHODS:A 3 mm by 3 mm specimen of canine oral mucosa was surgically excised and divided into epithelial and subepithelial tissues. Epithelial cells and fibroblasts were isolated and cultured separately. The proliferated epithelial cells were co-cultured on oriented collagen gels containing the proliferated fibroblasts for an additional two weeks. The organotypic cultured tissues were transplanted to the mucosa-deficient vocal folds. Two months after transplantation, vocal fold vibrations and morphological characteristics were observed. RESULTS:A tissue-engineered vocal fold mucosa, consisting of stratified epithelium and lamina propria, was successfully fabricated to closely resemble the normal layered vocal fold mucosa. Laryngeal stroboscopy revealed regular but slightly small mucosal waves at the transplanted site. Immunohistochemically, stratified epithelium expressed cytokeratin, and the distributed cells in the lamina propria expressed vimentin. Elastic Van Gieson staining revealed a decreased number of elastic fibers in the lamina propria of the transplanted site. CONCLUSION:The fabricated mucosa with autologous oral mucosal cells successfully restored the vocal fold mucosa. This reconstruction technique could offer substantial clinical advantages for treating intractable diseases such as scarring of the vocal folds.

At our institute, a chemoradioselection strategy has been used to select patients for organ preservation on the basis of response to an initial 30-40 Gy concurrent chemoradiotherapy (CCRT). Patients with a favorable response (i.e., chemoradioselected; CRS) have demonstrated better outcomes than those with an unfavorable response (i.e., nonchemoradioselected; N-CRS). Successful targeting of molecules that attenuate the efficacy of chmoradioselection may improve results. Thus, the aim of this study was to evaluate the association of a novel cancer stem cell (CSC) marker, CD44 variant 9 (CD44v9), with cellular refractoriness to chemoradioselection in advanced head and neck squamous cell carcinoma (HNSCC).Through a medical chart search, 102 patients with advanced HNSCC treated with chemoradioselection from 1997 to 2008 were enrolled. According to our algorithm, 30 patients were CRC following induction CCRT and 72 patients were N-CRS. Using the conventional immunohistochemical technique, biopsy specimens and surgically removed tumor specimens were immunostained with the anti-CD44v9 specific antibodies.The intrinsic expression levels of CD44v9 in the biopsy specimens did not correlate with the chemoradioselection and patient survival. However, in N-CRS patients, the CD44v9-positive group demonstrated significantly (P = 0.008) worse prognosis, than the CD44v9-negative group. Multivariate analyses demonstrated that among four candidate factors (T, N, response to CCRT, and CD44v9), CD44v9 positivity (HR: 3.145, 95% CI: 1.235-8.008, P = 0.0163) was significantly correlated with the poor prognosis, along with advanced N stage (HR: 3.525, 95% CI: 1.054-9.060, P = 0.0228). Furthermore, the survival rate of the CD44v9-induced group was significantly (P = 0.04) worse than the CD44v9-non-induced group.CCRT-induced CD44v9-expressing CSCs appear to be a major hurdle to chemoradioselection. CD44v9-targeting seems to be a promising strategy to enhance the efficacy of chemoradioselection and consequent organ preservation and survival.