Hypopharyngeal squamous cell carcinoma (HPSCC), an aggressive head and neck cancer with dismal prognosis, faces persistent chemoresistance to standard TPF (docetaxel, cisplatin, 5-fluorouracil) regimen. However, the immunological mechanisms underlying chemoresistance remain undefined. Here, we perform longitudinal single-cell RNA sequencing (scRNA-seq) profiling of paired pre-/post-TPF HPSCC specimens, mapping immune cell dynamics underlying chemoresistance. Our study identifies ZNF683+ natural killer (NK) cells as a gatekeeper of chemotherapy efficacy through integrated single-cell transcriptomics, spatial multiplex immunohistochemistry and functional validation. Moreover, pretreatment baseline enrichment of ZNF683+ NK cells predicts TPF response, while GZMK+CD8+ effector memory T cells function as the predominant immunologic effector to successful TPF intervention. Mechanistically, bioinformatics and in vitro coculture data reveal that ZNF683+ NK cells directly interact with CD8+ T cells, and drive an MHC-I-dependent licensing of polyfunctional GZMK+CD8+ effector memory T cells. Collectively, this NK-CD8+ axis provides a potential predictive biomarker and therapeutic target to overcome chemoresistance in patients with HPSCC.

BACKGROUND:To explore and compare the potential value of radiomics models based on contrast-enhanced computed tomography (CT) for noninvasive preoperative prediction of lymphovascular invasion (LVI) in laryngeal squamous cell carcinoma (LSCC). MATERIALS AND METHODS:This multicenter diagnostic study retrospectively enrolled patients with LSCC from three tertiary hospitals who underwent surgical treatment. Standardized preprocessing was performed on the CT images, followed by region-of-interest segmentation and extraction of traditional radiomics features and deep learning (DL) features. Features were selected using least absolute shrinkage and selection operator (LASSO) regression. Traditional radiomics models and deep learning radiomics (DLR) models were established using logistic regression, random forest, and multilayer perceptron algorithms, respectively. A transformer-based hybrid model was developed by integrating radiomics and DL features. The predictive performance of the three types of models was evaluated and compared using the area under the curve (AUC), decision curve analysis (DCA), sample probability distribution histograms, confusion matrices, calibration curves, net reclassification index (NRI), and integrated discrimination improvement (IDI). RESULTS:A total of 1024 patients were allocated to the training set (center1, n = 291), internal validation set ( n = 126), and external test sets (Center 2, n = 437; Center 3, n = 170). Three radiomics models and three DLR models were constructed, and the optimal performance was observed in the DLR_ Random Forest model (AUC: 0.812-0.867). The transformer hybrid model demonstrated superior predictive performance, with AUC values of 0.881, 0.843, 0.833, and 0.836 in the training, internal validation, and external test sets, respectively. DCA indicated a higher net benefit for the Transformer model, along with an improved NRI and IDI. CONCLUSION:Radiomics models based on CT images exhibit potential for noninvasive prediction of LVI in LSCC, with the transformer hybrid model achieving the highest diagnostic performance. This approach may provide clinicians with a preoperative decision support tool to optimize treatment strategies for patients with LSCC.

OBJECTIVE:The study aims to establish a pre-academic diagnostic tool based on deep learning and conventional radiomics features to guide the clinical decision-making of parapharyngeal space (PPS) tumors. METHODS:This retrospective study included 217 patients with PPS tumors, from two medical centers in China from March 1, 2011, to October 1, 2023. The study cohort was divided into a training set (n = 145) and a test set (n = 72). A deep learning (DL) model and conventional radiomics (Rad) model based on neck MRI were constructed to distinguish malignant tumors (MTs) and benign tumors (BTs) of PPS tumors. The deep learning radiomics (DLR) model which integrates deep learning and radiomics features was further developed. The area under the receiver operating characteristic curve (AUC), specificity, and sensitivity were used to evaluate model performance. Decision curve analysis (DCA) was applied to assess the clinical utility. RESULTS:Compared with the Rad and DL models, the DLR model showed excellent performance in this study, with the highest AUC of 0.899 and 0.821 in the training set and test set, respectively. The DCA curve confirmed the clinical utility of the DLR model in distinguishing the pathological types of PPS tumors. CONCLUSION:The DLR model demonstrated a high predictive ability in diagnosing MTs and BTs of PPS and could serve as a powerful tool to aid clinical decision-making in the preoperative diagnosis of PPS tumors. LEVEL OF EVIDENCE:3 Laryngoscope, 135:2275-2282, 2025.

BACKGROUND:The aim of this retrospective study was to compare the efficacy of transcervical (TC), endoscope-assisted transoral (TO), and endoscope-assisted TC for resection of retrostyloid space schwannomas. METHODS:The study included patients who underwent complete resection of schwannomas by only one surgical approach. The data we collected included tumor size, estimated blood loss, postoperative complications, and so on. Statistical analysis was performed using one-way analysis of variance and Fisher's exact test. RESULTS:The study collected 85 patients with tumors mostly located at the oropharyngeal level who were followed up 6 months at least. The results showed that endoscope-assisted TO had certain advantages over others. Additionally, the endoscope-assisted TO set the lowest incidence of neurological complications. CONCLUSION:Our findings demonstrate that for team with rich experience in the skull base surgery, endoscope-assisted TO is a superior option compared to the other two groups for resection of retrostyloid space schwannomas, with the better preservation of neurological function.

The biological functions of exosomes and microRNAs (miRs) in nasopharyngeal carcinoma (NPC) remain largely unexplored. Here, miR-197-3p was screened and identified, and whose level was reduced in serum and exosomes of patients with NPC. MiR-197-3p might be a good diagnostic and prognostic indicator. Our data showed that miR-197-3p expression was closely related to radioresistance, apoptosis, proliferation, migration, and survival of NPC. Inhibition of miR-197-3p expression in vitro could promote the proliferation and migration of NPC cells, while promotion of miR-197-3p expression in vivo could significantly inhibit the growth and enhance the radiosensitivity of NPC cells. From the perspective of mechanism, miR-197-3p could inhibit AKT/mTOR phosphorylation activation, inhibit an activated pathway of AKT/mTOR, target Heat Shock 70-kDa Protein 5(HSPA5) related to endoplasmic reticulum homeostasis, inhibit HSPA5-mediated autophagy, and reverse the radioresistance of NPC. Interestingly, exosomal miR-197-3p (EXO-miR-197-3p) reduced the proliferation and migration potential of NPC cells in vitro, and tumor growth and radioresistance of NPC cells in vivo. EXO-miR-197-3p inhibited NPC progression and radioresistance by regulating AKT/mTOR phosphorylation activation and HSPA5-mediated autophagy. In conclusion, our results highlight the potential of EXO-miR-197-3p as an effective radiosensitizer and therapeutic agent for refractory NPC.

Objectives To evaluate the effectiveness of machine learning models based on morphological magnetic resonance imaging (MRI) radiomics in the classification of parotid tumors. Methods In total, 298 patients with parotid tumors were randomly assigned to a training and test set at a ratio of 7:3. Radiomics features were extracted from the morphological MRI images and screened using the Select K Best and LASSO algorithm. Three-step machine learning models with XGBoost, SVM, and DT algorithms were developed to classify the parotid neoplasms into four subtypes. The ROC curve was used to measure the performance in each step. Diagnostic confusion matrices of these models were calculated for the test cohort and compared with those of the radiologists. Results Six, twelve, and eight optimal features were selected in each step of the three-step process, respectively. XGBoost produced the highest area under the curve (AUC) for all three steps in the training cohort (0.857, 0.882, and 0.908, respectively), and for the first step in the test cohort (0.826), but produced slightly lower AUCs than SVM in the latter two steps in the test cohort (0.817 vs. 0.833, and 0.789 vs. 0.821, respectively). The total accuracies of XGBoost and SVM in the confusion matrices (70.8% and 59.6%) outperformed those of DT and the radiologist (46.1% and 49.2%). Conclusion This study demonstrated that machine learning models based on morphological MRI radiomics might be an assistive tool for parotid tumor classification, especially for preliminary screening in absence of more advanced scanning sequences, such as DWI. Key Points • Machine learning algorithms combined with morphological MRI radiomics could be useful in the preliminary classification of parotid tumors. • XGBoost algorithm performed better than SVM and DT in subtype differentiation of parotid tumors, while DT seemed to have a poor validation performance. • Using morphological MRI only, the XGBoost and SVM algorithms outperformed radiologists in the four-type classification task for parotid tumors, thus making these models a useful assistant diagnostic tool in clinical practice.

Myeloid cells are a major heterogeneous cell population in the tumor immune microenvironment (TIME). Imbalance of myeloid response remains a major obstacle to a favorable prognosis and successful immune therapy. Therefore, we aimed to construct a risk model to evaluate the myeloid contexture, which may facilitate the prediction of prognosis and immune infiltration in patients with head and neck squamous cell carcinoma (HNSCC). In our study, six myeloid signature genes (including CCL13, CCR7, CD276, IL1B, LYVE1 and VEGFC) analyzed from 52 differentially expressed myeloid signature genes were finally pooled to establish a prognostic risk model, termed as myeloid gene score (MGS) in a training cohort and validated in a test cohort and an independent external cohort. Furthermore, based on the MGS subgroups, we were able to effectively identify patients with a poor prognosis, aggressive clinical parameters, immune cell infiltration status and immunotherapy response. Thus, MGS may serve as an effective prognostic signature and predictive indicator for immunotherapy response in patients with HNSCC.

Objectives/HypothesisTo analyze the clinical features, classification, and treatment of adult nasopharyngolaryngeal hemangioma (ANPLH).Study DesignRetrospective study.MethodsFrom February 2009 to May 2020, 101 patients with ANPLH were reviewed and analyzed.ResultsSymptoms of ANPLH were frequently displayed as abnormal pharyngeal sensation and functional defection. According to lesion location, ANPLH was divided into five categories including nasopharyngeal, oropharyngeal, hypopharyngeal, laryngeal, and mixed types. The mixed type constitutes the highest portion, and the nasopharyngeal type is the least in our cohort. Most lesions could resect through natural cavity under endoscopy. Patients with mixed lesions had a higher rate of postoperative recurrence and planned multiple surgeries. Acceptable but not severe intraoperative and postoperative complications occurred in our patient cohort.ConclusionsPatients with ANPLH are always symptomatic and even functional defective, which can be classified into five categories based on lesion location. For these patients, endoscopic surgery through natural cavity is recommended to remove lesions with fewer complications and favorable clinical outcomes.Level of Evidence4 Laryngoscope, 131:2724–2728, 2021

BACKGROUND:Tumor resistance to radiotherapy is one of the main obstacles to the clinical treatment of nasopharyngeal carcinoma (NPC). Improving the radiosensitivity of tumor cells has an important clinical significance in treatment of clinical NPC. This study aimed to identify that miR-138-1-3p as a novel therapeutic target in radioresistant NPC cells and found its targets, CRIPTO and the JAK2/STAT3 pathway.METHODS:Radioresistant C666-IR and HK-1R cells were derived from the NPC cell lines C666-1 and HK-1. The different microRNAs (miRNAs) and their targeting genes were analyzed between C666-1 and C666-IR cells using microarray bioinformatics. Western blot, qRT-PCR, gene transfection, Luciferase reporter assay, and confocal laser scanning microscopy were applied for the analysis of the different genes.RESULTS:MiR-138-1-3p was found to target CRIPTO, which involved in the epithelial-mesenchymal transition (EMT) and JAK2/STAT3 signaling pathways. The luciferase reporter assay confirmed that miR-138-1-3p targeted CRIPTO and downregulated the expression of CRIPTO. Furthermore, miR-138-1-3p affected the stability of the CRIPTO-GRP78 complex on the cell membrane and also reversed the radioresistant characteristics of NPC stem cells, which affected EMT and the JAK2/STAT3 signaling pathway.CONCLUSIONS:The miR-138-1-3p is a small molecule that can modulate radiosensitivity in the radioresistant C666-IR and HK-1R NPC cell lines by inhibiting EMT and targeting CRIPTO to reduce the activation of the JAK2/STAT3 pathway.

A novel fluorescent probe HPQB based on ESIPT and AIE was developed for the detection of H2O2 and glucose. HPQB was found with large Stokes shift (167 nm) characters. Probe HPQB exhibited high selectivity and excellent sensitivity for H2O2 and glucose. Moreover, HPQB was also employed as a fluorescent probe for imaging H2O2 in nasopharyngeal carcinoma (NPC) cells.

Bartter syndrome type IV, characterized by salt-losing nephropathies and sensorineural deafness, is caused by mutations of BSND or simultaneous mutations of both CLCNKA and CLCNKB. GJB2 is the primary causative gene for non-syndromic sensorineural deafness and associated with several syndromic sensorineural deafness. Owing to the rarity of Bartter syndrome, only a few mutations have been reported in the abovementioned causative genes. To investigate the underlying mutations in a Chinese patient with Bartter syndrome type IV, genetic analysis of BSND, CLCNKA, CLCNKB and GJB2 were performed by polymerase chain reaction and direct sequencing. Finally, double homozygous mutations c.22C > T (p.Arg8Trp) and c.127G > A (Val43Ile) were detected in exon 1 of BSND. Intriguingly, compound heterozygous mutations c.235delC (p.Leu79CysfsX3) and c.109G > A (p.Val37Ile) were also revealed in exon 2 of GJB2 in the same patient. No pathogenic mutations were found in CLCNKA and CLCNKB. Our results indicated that the homozygous mutation c.22C > T was the key genetic reason for the proband, and a digenic effect of BSND and GJB2 might contributed to sensorineural deafness. To our knowledge, it was the first report showing that the GJB2 gene mutations were detected in Bartter syndrome.

Autosomal dominant nonsyndromic hearing loss (ADNSHL/DFNA) is a highly genetically heterogeneous disorder. Hitherto only about 30 ADNSHL-causing genes have been identified and many unknown genes remain to be discovered. In this research, genome-wide linkage analysis mapped the disease locus to a 4.3 Mb region on chromosome 19q13 in SY-026, a five-generation nonconsanguineous Chinese family affected by late-onset and progressive ADNSHL. This linkage region showed partial overlap with the previously reported DFNA4. Simultaneously, probands were analyzed using exome capture followed by next-generation sequencing. Encouragingly, a heterozygous missense mutation, c.505G>A (p.G169R) in exon 3 of the CEACAM16 gene (carcinoembryonic antigen-related cell adhesion molecule 16), was identified via this combined strategy. Sanger sequencing verified that the mutation co-segregated with hearing loss in the family and that it was not present in 200 unrelated control subjects with matched ancestry. This is the second report in the literature of a family with ADNSHL caused by CEACAM16 mutation. Immunofluorescence staining and western blots also prove CEACAM16 to be a secreted protein. Furthermore, our studies in transfected HEK293T cells show that the secretion efficacy of the mutant CEACAM16 is much lower than that of the wild type, suggesting a deleterious effect of the sequence variant.

Waardenburg syndrome type IV (WS4) is a rare genetic disorder, characterized by auditory–pigmentary abnormalities and Hirschsprung disease. Mutations of the EDNRB gene, EDN3 gene, or SOX10 gene are responsible for WS4. In the present study, we reported a case of a Chinese patient with clinical features of WS4. In addition, the three genes mentioned above were sequenced in order to identify whether mutations are responsible for the case. We revealed a novel nonsense mutation, c.1063C>T (p.Q355*), in the last coding exon of SOX10. The same mutation was not found in three unaffected family members or 100 unrelated controls. Then, the function and mechanism of the mutation were investigated in vitro. We found both wild-type (WT) and mutant SOX10 p.Q355* were detected at the expected size and their expression levels are equivalent. The mutant protein also localized in the nucleus and retained the DNA-binding activity as WT counterpart; however, it lost its transactivation capability on the MITF promoter and acted as a dominant-negative repressor impairing function of the WT SOX10.

Objective To analyze the clinical phenotype of a pedigree with late-onset hereditary hearing loss,and investigate the disease-causing genes of this family. Methods The detailed medical history information of the participants in a large Hunan Province family with hereditary hearing loss were collected.Physical and augdiological examination were administered.Two individuals were carried out computed tomography(CT) scan of the temporal bone.The participants' information was sorted out and the genealogical tree map was charted.After reviewing the literature,we speculated on genes that could lead to deafness according to phenotype of this family.Some candidate genes coding region were amplified by polymerase chain reaction(PCR) using genomic DNA which was extracted from the proband's peripheral blood.PCR products were purified by exopeptidase.An ABI 3730XL genetic analyzer was used for direct sequencing,and data was analyzed using DNASTAR-Lasergene SeqMan Pro software. Results The family tree showed that this was an autosomal dominant non-syndromic deafness pedigree.Clinical features were remarkably similar among subjects.All of them were operated on for "buzz-like" tinnitus between 9 and 25 years old,and then for bilateral hearing loss.Pure tone audiometry examination showed the hearing loss began at the higher frequencies,then the lower frequencies with the aggravation of hearing loss. Conclusion Pedigree analysis suggested this family conformed to the characteristics of autosomal dominant hereditary pattern.The causative gene of this family needs more exploration.

Background and Aims. Bone morphogenetic proteins (BMPs) have recently been shown to be involved in the genesis and progression of a wide variety of carcinomas. The present study was undertaken to estimate the effect of BMP-4 on squamous cell carcinoma of the head and neck (SCCHN) in tissue and cell levels.Methods. In this study, immunohistochemistry, Western blotting and RT-PCR were utilized to detect the expression of BMP-4, Smad1 and phosphorylated Smad1 in SCCHN tissues or SCCHN cell lines. Those three proteins in tissues were further correlated with prognosis of SCCHN by Kaplan-Meier analysis. The epithelial-mesenchymal transition (EMT)-associated changes in SCCHN cells were detected after stimulation by human BMP-4 recombinant protein and knockdown of Smad1 gene. Meanwhile, the effect on invasiveness and migration was evaluated by invasion and scratch assays, respectively.Results. BMP-4 and p-Smad1 protein were overexpressed in SCCHN tissues with cervical lymph node metastasis, which was significantly higher than those without metastasis. The expression of BMP-4 and p-Smad1 protein was negatively correlated with the prognosis of SCCHN. BMP-4 promoted the invasiveness and migration through EMT, which was demonstrated by morphological alterations, loss of E-cadherin, increase of vimentin and activation of the Smad1 signal pathway. Knockdown of Smad 1 expression suppressed BMP-4 induced EMT in both cell lines and weakened the invasiveness and migration of Tu686 and Tu212 in vitro.Conclusions. Our results demonstrate that BMP-4 protein may contribute to the malignant metastasis of SCCHN, which presents as a novel prognostic marker and a potential therapeutic target for patients with SCCHN. (C) 2011 IMSS. Published by Elsevier Inc.

PURPOSE To investigate osteopontin (OPN) expression in human nasopharyngeal carcinoma (NPC) and evaluate its clinical significance in the disease. MATERIALS AND METHODS The expression of OPN mRNA in 44 frozen NPC tissue and 15 normal nasopharyngeal epithelium tissue (NNET) samples was examined by semi-quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). OPN protein expression in 67 paraffin-embedded NPC tissue and 21 NNET samples was detected by immunohistochemistry (IHC). In addition, OPN expression was investigated in 12 paired NPC and para-carcinoma tissue (PCT) samples by western blotting (WB). The association between the expression of OPN and the clinicopathologic parameters of NPC was evaluated. RESULTS Three different methods all showed that the expression of OPN mRNA or protein in NPC was significantly higher than in NNET or PCT (P = 0.000, 0.001, 0.000, respectively). After an examination by IHC, 88.1% (59/67) of NPC samples showed strong or moderate positive OPN staining and 28.6% (6/21) of NNET samples displayed a weak positive OPN staining. The staining of OPN in tumor cells was mainly localized to the cytoplasm. OPN expression in NPC was not related to patient age or sex (P > 0.05), but was significantly related to tumor size, regional lymph nodal metastasis, and NPC clinical stages (P < 0.05). CONCLUSIONS Our study demonstrated that OPN mRNA and protein overexpression in NPC may be important in the pathogenesis of the disease. It was strongly related to T stage, N stage and clinical stages of NPC, suggesting that OPN may be involved in NPC metastasis and progression.