Background: B7-H3 (CD276) is an immune checkpoint molecule overexpressed in head and neck squamous cell carcinoma (HNSCC) and associated with immunosuppression, stemness, and poor prognosis. While a promising target for chimeric antigen receptor (CAR) T-cell therapy, the clinical translation of B7-H3 CAR-T cells is hampered by the limited persistence induced by high immunogenicity or exhaustion associated with excessive tonic signaling. Methods: We characterized B7-H3 expression and its associated tumor microenvironment in HNSCC using TCGA data, patient specimens, and cell lines. We constructed B7-H3 CARs incorporating either the murine 8H9 or human MGA271 scFv and evaluated their function. To mitigate the tonic signaling of the MGA271 CAR, we performed structure-guided mutagenesis to reduce positive surface charge patches on the scFv. The resulting charge-optimized CARs were assessed for tonic signaling, exhaustion markers, and antitumor efficacy in vitro and in vivo. Results: B7-H3 was broadly overexpressed in HNSCC and defined an immunosuppressive tumor microenvironment characterized by CD8+ T-cell exclusion and predicted resistance to PD-1 blockade. The human-derived MGA271 CAR-T cells exhibited severe tonic signaling, evidenced by elevated basal CD69 and PD-1 expression, leading to premature exhaustion and impaired proliferation. Computational analysis revealed prominent positive charge patches on the MGA271 scFv. Charge-optimized variants (MGA271M2) demonstrated reduced surface charge, which effectively alleviated tonic signaling, decreased exhaustion markers, and enhanced proliferative capacity. Crucially, MGA271M2 CAR-T cells retained potent cytotoxicity in vitro and mediated superior tumor control and persistence in HNSCC xenograft models compared to the wild-type CAR. Conclusion: Our study unequivocally establishes B7-H3 as a compelling therapeutic target in immunosuppressive and aggressive HNSCC. Moreover, we pioneer a rational charge-optimization strategy that successfully mitigates CAR-T cell exhaustion, thereby creating a potent and persistent B7-H3-targeted therapy with broad implications for solid tumor treatment.

OBJECTIVES:To make an anatomical comparison between porcine and human laryngeal structures, and evaluate suitability for microsurgical training in the porcine larynx. METHODS:Thirty Landrace pig larynges underwent muscle-dissection to expose thyroid cartilage, cricoid cartilage, and intrinsic laryngeal structures. Quantitative analysis was conducted comparing porcine cartilage dimensional data with human specimens. Histological evaluation of vocal fold architecture was performed using hematoxylin-eosin (H-E) and Masson staining. The laryngeal internal structures were exposed using a laryngeal fixation stent and a surgical microscope to evaluate the accessibility for microsurgical operations. The applicability of cold dissection, CO2 laser, fiber-contact laser, and plasma scalpel were evaluated for microsurgical training. RESULTS:The average size of the thyroid cartilage in 6-month-old pigs is 62.8 mm (length) × 42.0 mm (width), equivalent to 110.5-136.2% of the size of human larynx. Histological staining results reveal distinct structural differences between the superior and inferior vocal folds. The inferior vocal fold contains muscle bundle, while the superior vocal fold is rich in connective tissue and lacks muscle bundle. Under a surgical microscope, the superior and inferior vocal folds and the laryngeal ventricle can be clearly exposed. Common clinical techniques such as vocal mucosal dissection, vocal fold resection, and microsurgical suturing can be precisely simulated in the porcine larynx. CONCLUSION:The structure of the porcine larynx was similar to that of the human larynx. The larger overall structure was more conducive to the laryngeal microsurgery training of otolaryngologists.

OBJECTIVES:To establish a combined classification based on intrapapillary capillary loops (IPCLs) and morphological characteristics to improve the accuracy of pathological prediction of vocal fold leukoplakia (VFL). MATERIALS AND METHODS:A prospective research was conducted of VFL patients diagnosed by IPCLs and morphology. The VFL cases were classified as Type I-III based on IPCLs and morphological characteristics. Type I referred to VFL with dendritic vessels but not IPCLs. Type II defined VFL without any IPCLs or vessels and classified by morphology into two subtypes as nonrough Type IIa and rough Type IIb. Type III referred to VFL with IPCLs and classified into two subtypes as Type IIIa with small IPCLs and Type IIIb with large IPCLs or vascular distortion in or around lesions. Predicting pathology accuracy was analyzed. RESULTS:One hundred and eighty-two eligible patients were recruited. The prediction accuracy rates of VFL pathology were 81.5% according to the 2019 Ni classification. The combined classification included four cases of Type I, 28 Type IIa, 35 Type IIb, 56 Type IIIa, and 59 Type IIIb VFLs. The prediction accuracy rate of combined classification ranged from 95.1% to 97.3% in three observers. The average sensitivity, specificity, positive predictive value, and negative predictive value were 97.8%, 86.2%, 97.4%, and 88.2%, respectively. The interobserver agreement varied from 84.1% to 94.0%; the mean area under curve of receiver operating curve analysis was 0.954. CONCLUSIONS AND SIGNIFICANCE:The new combined classification based on IPCLs and morphological characteristics could predict the pathology of VFL accurately. LEVEL OF EVIDENCE: 4:

Major challenges of developing CAR‑T cell therapy for head and neck squamous cell carcinoma (HNSCC) include identifying a robust tumor antigen and a suitable CAR design. Here, we validate chondroitin sulfate proteoglycan 4 (CSPG4) as a highly expressed, prognostic antigen in HPV‑negative HNSCC that drives tumor proliferation. By grafting the murine single-chain variable fragment (scFv) 763.74 complementarity determining regions (CDRs) onto a human antibody framework engineered to minimize surface positive‑charge patches and immunogenic epitopes, we generated humanized CSPG4 (CSPG4Hu) CAR‑T cells with reduced tonic signaling and alleviated exhaustion. Transcriptomic and metabolic profiling reveal that this biophysical refinement reprograms CAR-T cells away from a glycolytic, terminal exhaustion state toward a PI3K/Akt-driven stem-like state. Consequently, CSPG4Hu CAR-T cells demonstrate superior persistence and potent antitumor efficacy across systemic xenograft and patient-derived xenograft models. Our study establishes a rational engineering framework that links biophysical CAR design to transcriptomic and metabolic rejuvenation, offering a promising therapeutic candidate for advanced HNSCC.

As narrow band imaging（NBI） technology and research for vocal fold leukoplakia have advanced, the diagnostic and therapeutic paradigms for this condition have evolved significantly. This paper provides insight into the challenges of clinical management of vocal fold leukoplakia, drawing on our empirical findings to clarify the etiology, differential diagnosis, pathological grading, morphological characteristics, and recent therapeutic developments of this condition.

Vocal fold leukoplakia (VFL), a precancerous lesion of the larynx, is characterized by white plaques on the vocal fold mucous membrane. Currently, there are no reliable biomarkers to predict the recurrence and malignant transformation of VFL. Considering chondroitin sulfate proteoglycan 4 (CSPG4) as a biomarker for malignant tumors such as laryngeal squamous cell carcinoma (LSCC), we conducted this cohort study to evaluate the prognostic influence of CSPG4 expression on VFL patients. This study included 44 patients with surgical intervention of VFL. CSPG4 expression in VFL were observed using immunohistochemistry, and the relationship between the prognosis of VFL patients and CSPG4 expression were investigated using Spearman correlation analysis and multiple Cox regression models. During a median follow-up of 89.3 months, recurrence occurred in 19 postsurgical VFL patients, and malignant transformation occurred in 10 patients. The recurrence and malignant transformation group showed significant differences in the H-SCORE of CSPG4 (P = 0.005 and P = 0.045) compared to the the non-recurrence group and the non-malignant transformation group. And the CSPG4 expression level was positive correlation with recurrence and malignant transformation (0.306 or 0.416, spearman correlation confident). Receiver operating characteristic curve analysis indicated that CSPG4 H-SCORE has predictive value for poor prognosis (areas under the curve, 0.742 and 0.710; cut-off points, 82.5 and 177.5, respectively). Multiple Cox regression analysis revealed a statistically significant effect of lesion size (P = 0.014) and CSPG4 H-SCORE (P = 0.005) on recurrence as well as pathological type (P = 0.001) and CSPG4 H-SCORE stratified by 177.5 (P = 0.039). Our survival data indicated that higher CSPG4 expression was associated with a shorter time of recurrence (P = 0.003) and tumorigenesis (P<0.001). CSPG4 overexpression indicates higher risks and shorter time of postoperative VFL recurrence and tumorigenesis. Detection of CSPG4 expression in VFL may be a novel approach to assess the outcome of VFL patients.

BACKGROUND:The reconstruction of hypopharyngeal tumor defect after tumorectomy is essential. Artificial materials play an important role in the case of extensive defects and a relative lack of healthy tissue. This study aims to investigate the role of acellular dermal matrix (ADM) in repairing defects following hypopharyngeal carcinoma resection. METHODS:A retrospective analysis was conducted on 280 patients with hypopharyngeal carcinoma. 143 patients accepted ADM-assisted reconstruction while 137 patients didn't. Data on postoperative deglutition function, respiratory function, and complications were collected and analyzed. RESULTS:No immune rejection response was observed. In partial laryngectomy, ADM significantly induces the time for hospitalization, decannulation, and oral food intake compared to the control group (p < 0.05). In total laryngectomy, ADM has advantages in terms of the number of individuals with good deglutition function (p < 0.05). There is no significant difference in pharyngocutaneous fistula. CONCLUSIONS:ADM is a promising biomaterial for reconstruction in hypopharyngeal carcinoma. It offers advantages such as a high success rate, good graft survival, accessibility, minimal immunogenicity, and simplified surgical procedures.

It was aimed to explore an alternative solution to manage T1 glottic cancer patients. One hundred seventy-three patients were enrolled. Seventy-eight cases were treated with type I and proved to be microinvasion cancer. Of them, 21 willingly selected followed up and 57 were further undergone type II. Ninety-five underwent type III and confirmed to be superficial invasion cancer in which Reinke's space was invaded. Five(23.8%), four(7.0%) and nine (9.5%)cases separately relapsed in type I, II and III. No significant difference was observed (p>0.05). 5/60, 5/11 and 8/102 patients respectively replaced in negative, positive and uncertain margin groups. There were remarkable differences among them (p<0.05). The recurrence rate is still low for patients with early glottic cancer undergone vocal cord resection using CO2 laser, and the appropriate surgical method should be selected for a diverse range of glottic cancer to avoid overtreatment. Patients with positive incisal margins have a relatively high recurrence rate and should take this situation seriously.

Objectives: Polysomnography was class I test for who was suspected of obstructive sleep apnea (OSA) which would cost lots of time and money. This study aimed to develop a nomogram model mainly based on oxygen and blood routine indicators to predict OSA. Methods: We retrospectively analyzed 685 patients with suspected OSA at our hospital. Multivariate analysis was used to construct a nomogram. The performance of the nomogram was assessed using calibration and discrimination. Results: The multivariate analysis identified age, gender, body mass index, mean pulse oxygen saturation, percent nighttime with oxygen saturation less than 90%, red blood cell, hematocrit, and red blood cell distribution width SD as significant factors (P < .05). A nomogram was created for the prediction of OSA using these clinical parameters and was internally validated using a bootstrapping method. Our nomogram model showed good discrimination and calibration in terms of predicting OSA, and had a C-index of 0.935 [95% confidence interval (CI), 0.917-0.954] according to the internal validation. Discrimination and calibration in the validation group were also good (C-index, 0.957; 95% CI, 0.930-0.984). Conclusion: The newly developed nomogram can effectively help physicians make better clinical decisions, which may save a lot of time and costs.

Objectives: To investigate the pathological contribution of vocal fold leukoplakia (VFL) of type II in narrow-band imaging (NBI) classification and morphological characteristics to improve pathological prediction. Material and Methods: The 59 VFL patients with type II in 2019 Ni classification in NBI were included. The pathological reports were collected and divided following 2005 WHO Blue Book. Low-risk VFL contained non-, mild, moderate dysplasia, high-risk VFL included severe dysplasia. The morphological classification and laryngoscopic scoring system were employed to evaluate leukoplakia for pathological prediction. Results: The pathologies contained 1 case of leukoplakia with non-dysplasia, 12 of mild dysplasia, 15 of moderate dysplasia, 8 of severe dysplasia, and 23 of carcinoma. The 30 smooth VFL contained 1 non-dysplasia, 12 mild dysplasia, 14 moderate dysplasia, 2 severe dysplasia, and 1 carcinoma. The 29 rough cases included 1 moderate dysplasia, 6 severe dysplasia, and 22 carcinomas. Laryngoscopic scoring system revealed irregular texture, large size, and thick lesion as factors in relationship with high-risk leukoplakia in univariate ( P = .002, <.001, <.001) and multivariate ( P = .025, .002, .016) analysis, irregular texture was the most accurate predictor of high-risk VFL pathology. Conclusions and Significance: The pathologies of VFL with type II in NBI classification were hard to be predicted. Morphological irregular/rough texture contributed to predict high-risk pathology in leukoplakia.

IMPORTANCE:Patients who undergo total laryngectomy lose the voice function permanently. It is important to reconstruct the voice function of the patients after total laryngectomy. OBJECTIVE:To explore a novel method for voice rehabilitation by suturing infrahyoid myocutaneous flaps to the hypopharyngeal and esophageal serosas after total laryngectomy and investigate its clinical efficacy. DESIGN:Clinical study (retrospective study). PARTICIPANTS:Twenty patients with advanced laryngocarcinoma who underwent total laryngectomy. METHODS:Following total laryngectomy, the pharyngeal cavity was formed via layer-by-layer suturing, and the infrahyoid myocutaneous flap was sutured to the common wall of hypopharyngeal and esophageal serosas to create a voice tube. RESULTS:Flap failure was not observed in any patient. All patients were able to vocalize after surgery. The voice after the reconstruction was smooth and natural. However, 3 patients were unable to vocalize gradually. Two patients experienced aspiration, requiring the removal of the voice tube, while 3 patients exhibited mild aspiration, which could be overcome by pressing the larynx on the voice tube surface. CONCLUSIONS:Using infrahyoid myocutaneous flaps with the common wall of the hypopharyngeal and esophageal serosas was effective in rehabilitating voice after total laryngectomy. Future clinical studies are necessary to validate the effectiveness of this technology for voice rehabilitation.

Objective To analyse the comparative clinical outcomes and clinicopathological significance of vocal fold leukoplakia lesions treated by appearance classification and traditional methods. Method A total of 1442 vocal fold leukoplakia patients were enrolled. Group A patients were treated according to appearance classification and Group B patients were treated according to traditional methods. Results In Group A, 24.4, 14.9 and 60.6 per cent of patients had grade I, II and III dysplasia, respectively. Grade I dysplasia (63.4 per cent) was more than twice as frequent in Group B patients than in Group A patients, while grade II dysplasia (20.4 per cent) and grade III dysplasia (16.2 per cent) were significantly less frequent in Group B patients than in Group A patients ( p = 0.000). There was a significant correlation between vocal fold leukoplakia appearance and the degree of dysplasia ( p = 0.000). The recurrence and malignant transformation rates (17.6 and 31 per cent, respectively) in Group B were significantly greater than those in Group A (10.8 and 25.9 per cent, respectively) ( p = 0.000). Conclusion Vocal fold leukoplakia appearance classification is useful for guiding treatment decision-making and could help to improve therapeutic accuracy.

Objectives: To evaluate the possible risk factors of obstructive sleep apnea (OSA) and its impact on the health-related quality of life (HRQoL) in patients with partial laryngectomy for laryngeal cancer. Methods: A cross-sectional method was used to carry out this study. Patients who underwent partial laryngectomy for laryngeal cancer completed overnight polygraphy (PG) home sleep tests and quality of life questionnaires. The Medical Outcome Study 36-item Short-Form Health Survey (SF-36) questionnaire was used to investigate the factors influencing HRQoL. Results: A total of 59 patients completed the PG tests and quality of life questionnaires, with 74.6% demonstrating evidence of OSA. There were significant differences in tumor area and neck dissection between OSA group and non-OSA groups. Based on sleep-related parameters, patients were divided into cluster 1 (n = 14) and cluster 2 (n = 45) using principal component analysis combined with K-means clustering. Two clusters had significantly different scores of body pain, general health, and health transition in SF-36 domains. Independent factors associated with general health were identified as tobacco use (OR = 4.716), alcohol use (OR = 3.193), and OSA-related condition (OR = 11.336). Conclusions: Larger tumor area and neck dissection might be associated with an increased risk for developing OSA in patients with partial laryngectomy for laryngeal cancer. OSA partially mediated the effect on physical health, including body pain, general health, and health transition. It is important to be aware of the potential impact of OSA on diminished HRQoL of these patients.

BACKGROUND/AIMS/OBJECTIVES:To investigate the treatment for adult glottic stenosis using CO2 laser surgery combined with a self-made laryngeal dilator.MATERIAL AND METHODS:A retrospective analysis was performed on 18 patients with glottic stenosis who were treated using CO2 laser surgery combined with a self-made laryngeal dilator in our hospital from January 2018 to December 2020.RESULTS:4 cases were caused by trauma and one by laryngophthisis. Laryngeal stenosis occurred in 4 and 9 patients respectively after CO2 laser surgery and open partial laryngectomy. Of them, one patient underwent postoperative radiotherapy. All patients were treated through CO2 laser surgery combined with a self-made laryngeal dilator under general anesthesia. 3-6 months later, the dilator was removed. Inflammation, ulceration and granulation were observed surrounding the dilator. But these complications would be cured and respiration was not affected. Finally, four patients could not be extubated and the decannulation rate achieved 78%. All patients successfully decannulated could normally intake. 13 cases had good voice quality and only one patient pronounced hoarsely.CONCLUSIONS AND SIGNIFICANCE:It is demonstrated that the application of CO2 laser surgery combined with a laryngeal self-made dilator is feasible and effective for the treatment with adult glottic stenosis.

Objectives To investigate the clinical outcomes of morphology-guided surgical treatment for adult laryngeal and hypopharyngeal hemangioma. Materials and methods Patients diagnosed with adult laryngeal and hypopharyngeal hemangioma by outpatient laryngoscopy and received surgical treatments were retrospectively included. Size and morphology were classified to guide the choose of surgical procedures including CO2 laser excision, CO2 coagulation + bleomycin injection and potassium-titanyl-phosphate (KTP) laser + bleomycin injection. The baseline information and pre-, postoperative laryngoscopies were collected, the outcomes were followed up and analyzed. Results A total of 68 patients were included, 7 cases with exophytic and non-bulky hemangioma received CO2 laser excision, 20 cases received CO2 laser coagulation + bleomycin injection and 25 cases with KTP + bleomycin injection, another 16 cases with pure bleomycin injection were included as comparison. There was no difference of baseline analysis among the groups (P > = 0.05). Positive outcomes were achieved in most cases of CO2 excision (100%), CO2 coagulation + bleomycin (90.0%) and KTP + bleomycin (84.0%) groups, which were better than pure bleomycin (56.3%, p = 0.001). CO2 or KTP, associated to Bleomycin, gave same results on same lesions (P = 0.132). Conclusions and significanceMorphology of hemangioma may help in the selection of surgical procedures. CO2 coagulation can be used as a potential substitute for KTP in the treatment of adult laryngeal and hypopharyngeal hemangioma.

BACKGROUND:The treatment of refractory hypopharyngeal corrosive strictures remains challenging nowadays. OBJECTIVES:To introduce a designed ladder-shaped stent for the treatment of refractory hypopharyngeal corrosive strictures. MATERIAL AND METHODS:Patients with refractory hypopharyngeal corrosive strictures treated in our hospital between 2018 to 2022 were recruited in this study. All the patients had a dysphasia score of 4 and four patients had a tracheostomy preoperatively. A designed ladder-shaped stent was placed in the hypopharyngeal area after laser treatment to maintain the opening. This stent was kept for 3-6 months. All the patients were followed-up for at least 6 months after stent removal. RESULTS:Six patients were recruited including two males, three females and one girl. All the patients were tolerable with the stent. There were no immediate or long-term complications. Patients were able to swallow a soft diet after the procedure. No tracheostomy was needed. CONCLUSION:This use of a ladder-shaped stent is helpful in the treatment of hypopharyngeal strictures after laser treatment.

Gastric cancer (GC) is a major health burden worldwide, but our understanding of GC is limited, and the prognosis is poor. Novel therapeutic strategies and biomarkers are urgently needed to improve GC patient outcomes. Previously, we identified PFDN2 as a novel key gene in gastric cancer based on its differential expression between cancer and normal tissues. However, the role and underlying mechanisms of PFDN2 in GC remain elusive. In this article, we demonstrated that PFDN2 is highly expressed in GC and that upregulation of PFDN2 is associated with the progression of GC. We further found that PFDN2 could promote cell cycle progression by promoting MYBL2 expression. Mechanistically, we demonstrated that PFDN2 could upregulate MYBL2 expression by facilitating the nuclear translocation of hnRNPD, and thus promoting MYBL2 transcriptional program. In conclusion, we found that PFDN2 promotes cell cycle progression via the hnRNPD-MYBL2 axis and may serve as a potential biomarker and therapeutic target for GC.