Patterns of cervical lymph node involvement in mono-focal papillary thyroid carcinoma patients with different capsular status: A multi-center analysis.

OBJECTIVE: The characteristics of cervical lymph node involvement in papillary thyroid carcinoma (PTC) patients with different degree of capsular invasion remains unclear, especially for those with mono-focal lesion who have traditionally been considered as low neck metastasis risk subgroup. STUDY DESIGN: Retrospective cohort study. SETTING: Three academic teaching hospital. METHODS: A total of 1276 mono-focal PTC patients were retrospectively analyzed. RESULTS: Mono-focal PTC patients with extrathyroidal extension (ETE) showed significantly higher central lymph node metastasis (CLNM) rate than those without. For patients with no gross ETE (gETE), those with minimal ETE (mETE) also showed more commonly CLNM than those with encapsulated lesions. However, the lateral lymph node metastasis (LLNM) rates of patients with mETE and encapsulated tumors were comparable, both lower than that of patients with gETE. Age ≥40, male, and MTD ≥0.5 cm were identified as independent risk factors of CLNM for those with encapsulated tumors and were enrolled for creating a prediction model. In terms of LLNM, only MTD ≥1.0 cm was confirmed as independent risk factors of LLNM for patients with positive gETE. CONCLUSIONS: The presence and degree of ETE may have different effects on the risk of central and lateral lymph node metastasis. gETE demonstrates a strong correlation with both CLNM and LLNM while mETE is only associated with CLNM in mono-focal PTC patients. A comprehensive model is established in the aim of predicting neck involvement according to the capsular status and the corresponding stratified risk factors, which may aid clinical decision-making for the management of neck regions.

Peripheral lymphocyte count as an indicator of radiotherapy effectiveness in hypopharyngeal squamous cell carcinoma.

OBJECTIVES: To explore the predictive significance of baseline absolute peripheral lymphocyte counts (ALC) in the effectiveness of radiation in hypopharyngeal squamous cell carcinoma (HPSCC) patients. DESIGN, SETTING, AND PARTICIPANTS: A retrospective study of pathologically confirmed HPSCC patients who had definitive radiation between January 2020 and January 2022 at Fudan University Eye and ENT Hospital. The routine blood results of patients were obtained to determine if the baseline ALC was connected with the response to radiation. The receiver operator characteristic (ROC) curve and LASSO-based Cox regression were employed to assess the predictive value of ALC for the efficacy of radiotherapy (RT). MAIN OUTCOME MEASURES AND RESULTS: RT induced a considerable drop in ALC and the level of ALC did not revert to the baseline values 1 year after radiation. The baseline level of ALC was higher in patients who met complete response after RT. The baseline ALC and monocyte counts demonstrated the predictive value of radiation effectiveness and ALC was an independent predictor. CONCLUSION: In HPSCC, lymphocytes were sensitive to radiation and reduced significantly during RT. The baseline ALC might be regarded as a predictive indicator of the effectiveness of RT.

Increased tumor-infiltrating plasmacytoid dendritic cells express high levels of PD-L2 and affect CD8+ T lymphocyte infiltration in human laryngeal squamous cell carcinoma.

The infiltration and prognostic significance of tumor-infiltrating plasmacytoid dendritic cells (TI-pDC) have been elucidated in various human solid cancers. However, the infiltrating patterns and functional importance of TI-pDC in laryngeal squamous cell carcinoma (LSCC) remain unknown. In this study, flow cytometric analyses were conducted to characterize the infiltration of dendritic cells and T lymphocytes, along with their respective subgroups in tumor tissues (TT), para-carcinoma tissues (PT), and peripheral blood (PB) from LSCC patients. Immunohistochemical staining for CD4 and CD8, as well as immunofluorescence staining for CD123, were performed on serial tissue sections to investigate the co-localization of TI-pDC and tumor-infiltrating T lymphocytes (TIL) within the tumor microenvironment (TME). Our results demonstrated significantly lower percentages of all three DC subsets in PB compared to TT and PT. Notably, the pDC percentage was markedly higher in TT than in PT. Moreover, TI-pDC percentage was significantly elevated in N+ stage patients compared to those with N0 stage. The results of survival analysis consistently demonstrated that high levels of TI-pDC infiltration were indicative of a poor prognosis. Further investigation revealed a significant negative correlation between TI-pDC and CD8+ TILs; notably, pDCs expressed an inhibitory surface molecule PD-L2 rather than PD-L1 within PT. Collectively, our findings suggest that increased TI-pDC is associated with adverse outcomes in LSCC patients while exhibiting an inhibitory phenotype that may play a crucial role in suppressing CD8+ TILs within LSCC tumors. These results highlight the potential therapeutic strategy targeting PD-L2+ pDCs for immunotherapies against LSCC.

Collaborative Modality Fusion for Mitigating Language Bias in Visual Question Answering.

Language bias stands as a noteworthy concern in visual question answering (VQA), wherein models tend to rely on spurious correlations between questions and answers for prediction. This prevents the models from effectively generalizing, leading to a decrease in performance. In order to address this bias, we propose a novel modality fusion collaborative de-biasing algorithm (CoD). In our approach, bias is considered as the model's neglect of information from a particular modality during prediction. We employ a collaborative training approach to facilitate mutual modeling between different modalities, achieving efficient feature fusion and enabling the model to fully leverage multimodal knowledge for prediction. Our experiments on various datasets, including VQA-CP v2, VQA v2, and VQA-VS, using different validation strategies, demonstrate the effectiveness of our approach. Notably, employing a basic baseline model resulted in an accuracy of 60.14% on VQA-CP v2.

Prognostic significance and immune escape implication of tumor-infiltrating neutrophil plasticity in human head and neck squamous cell carcinoma.

Tumor-infiltrating neutrophils play a crucial role in the progression of head and neck squamous cell carcinoma (HNSCC). Here, we aimed to statistically quantify the plasticity of HNSCC-infiltrating N2/N1 neutrophils and examine its impacts on survival and immune infiltration landscape. A retrospective study of 80 patients who underwent curative surgical resection for HNSCC between 2014 and 2017 was conducted in this study. HNSCC-infiltrating neutrophil phenotypes were classified using immunofluorescence staining, and the N2/N1 neutrophil plasticity was evaluated via the ratio of N2/N1 neutrophils. We then assessed the correlations between N2/N1 neutrophil plasticity, clinicopathological characteristics, and immune infiltration landscape using rigorous statistical methods. Infiltration variations of N1 and N2 neutrophils were observed between the tumor nest (TN) and tumor stroma (TS), with TN exhibiting higher N2 neutrophil infiltration and lower N1 neutrophil infiltration. High ratios of N2/N1 neutrophils were correlated with advanced TNM stage, large tumor size and invasion of adjacent tissue. High infiltration of N2 neutrophils was associated with decreased overall and relapse-free survival, which were opposite for N1 neutrophils. The independent prognostic role of N2/N1 neutrophil plasticity, particularly within the TN region, was confirmed by multivariate analyses. Moreover, the ratio of N2/N1 neutrophils within the TN region showed correlations with high CD8+ T cells infiltration and low FOXP3+ Tregs infiltration. We identify HNSCC-infiltrating N2/N1 neutrophil plasticity as a crucial prognostic indictor which potentially reflects the tumor microenvironment (TME) and immune escape landscape within HNSCC tissues. Further investigations and validations may provide novel therapeutic strategies for personalized immunomodulation in HNSCC patients.

Establishment and characterization of a novel hypopharyngeal squamous cell carcinoma cell line CZH1 with genetic abnormalities.

Hypopharyngeal squamous cell carcinoma (HPSCC) has the worst prognosis among head and neck squamous cell carcinomas. The lack of available tumor cell lines poses a significant obstacle to the development of efficient treatments for HPSCC. In this study, we successfully established a novel cell line, named CZH1, from the postcricoid region of a Chinese male patient with a T3N0M0 HPSCC. Short tandem repeat analysis confirmed the uniqueness of CZH1. The cell line was characterized by its phenotypes, biomarkers, and genetics. Importantly, CZH1 cells retained the typical features of epithelial malignancy, similar to the primary tumor tissue. Furthermore, CZH1 demonstrated a greater capacity for invasion and increased susceptibility to irradiation in comparison to FaDu, which is the most commonly used HPSCC cell line. Whole-exome sequencing analysis revealed that CZH1 cells had typical genomic features of HNSCC, including mutations of TP53 and amplifications of multiple transcripts. Therefore, our newly developed CZH1 cell line could serve as an efficient tool for the in vitro investigation of the etiology, pathogenesis, and preclinical treatment of HPSCC.

A novel two-way rebalancing strategy for identifying carbonylation sites.

BACKGROUND: As an irreversible post-translational modification, protein carbonylation is closely related to many diseases and aging. Protein carbonylation prediction for related patients is significant, which can help clinicians make appropriate therapeutic schemes. Because carbonylation sites can be used to indicate change or loss of protein function, integrating these protein carbonylation site data has been a promising method in prediction. Based on these protein carbonylation site data, some protein carbonylation prediction methods have been proposed. However, most data is highly class imbalanced, and the number of un-carbonylation sites greatly exceeds that of carbonylation sites. Unfortunately, existing methods have not addressed this issue adequately. RESULTS: In this work, we propose a novel two-way rebalancing strategy based on the attention technique and generative adversarial network (Carsite_AGan) for identifying protein carbonylation sites. Specifically, Carsite_AGan proposes a novel undersampling method based on attention technology that allows sites with high importance value to be selected from un-carbonylation sites. The attention technique can obtain the value of each sample's importance. In the meanwhile, Carsite_AGan designs a generative adversarial network-based oversampling method to generate high-feasibility carbonylation sites. The generative adversarial network can generate high-feasibility samples through its generator and discriminator. Finally, we use a classifier like a nonlinear support vector machine to identify protein carbonylation sites. CONCLUSIONS: Experimental results demonstrate that our approach significantly outperforms other resampling methods. Using our approach to resampling carbonylation data can significantly improve the effect of identifying protein carbonylation sites.

High Expression of Tumor HLA-DR Predicts Better Prognosis and Response to Anti-PD-1 Therapy in Laryngeal Squamous Cell Carcinoma.

BACKGROUND: HLA-DR is expressed in epithelial and several types of tumor cells. However, the correlation between tumor-expressed HLA-DR (teHLA-DR) and patient outcome as well as its regulation on the tumor microenvironment (TME) of laryngeal squamous cell carcinoma (LSCC) are yet to be elucidated. METHODS: Hematoxylin and eosin (HE) staining were performed to define the tumor nest and stroma of LSCC tissue microarrays. teHLA-DR tumor cell, CD4+ and CD8+ tumor-infiltrating T lymphocytes (TITLs) were obtained and analyzed through double-labeling immunofluorescence and immunohistochemical staining. The recurrence-free (RFS) and overall survival (OS) curves were plotted using the Kaplan-Meier method and tested by the log-rank test method. Expression of teHLA-DR+ tumor cells and infiltration of T lymphocytes and their corresponding subgroups were analyzed by flow cytometry using fresh LSCC tissue samples. RESULTS: Our research discovered elevated expressions of multiple MHC-II-related genes in tumor compared to the adjacent normal tissue samples of LSCC patients. We also found that patients in the teHLA-DR high-expression group (teHLA-DRhigh) tend to have less tumor recurrence and better survival outcomes compared to those in the teHLA-DRlow group. Intriguingly, teHLA-DR+ tumor cells had significantly higher PD-L1 and PD-L2 expression and their TME showed increased infiltrated T lymphocytes (TITLs). Flow cytometry analysis and IHC staining indicated that CD4+ TITLs but not CD3+ total TITLs or CD8+ TITLs were significantly enriched in teHLA-DR+ tumors. CONCLUSIONS: teHLA-DR may be a predictive marker for favorable prognosis and response to anti-PD-1/PD-L1 therapy of LSCC, possibly due to the increased CD4+ TITLs in the TME.

DeepSG2PPI: A Protein-Protein Interaction Prediction Method Based on Deep Learning.

Protein-protein interaction (PPI) plays an important role in almost all life activities. Many protein interaction sites have been confirmed by biological experiments, but these PPI site identification methods are time-consuming and expensive. In this study, a deep learning-based PPI prediction method, named DeepSG2PPI, is developed. First, the protein sequence information is retrieved and the local context information of each amino acid residue is calculated. A two-dimensional convolutional neural network (2D-CNN) model is employed to extract features from a two-channel coding structure, in which an attention mechanism is embedded to assign higher weights to key features. Second, the global statistical information of each amino acid residue and the relationship graph between the protein and GO (Gene Ontology) function annotation are built, and the graph embedding vector is constructed to represent the biological features of the protein. Finally, a 2D-CNN model and two 1D-CNN models are combined for PPI prediction. The comparison analysis with existing algorithms shows that the DeepSG2PPI method has better performance. It provides more accurate and effective PPI site prediction, which will be helpful in reducing the cost and failure rate of biological experiments.

CD206+ tumor-associated macrophages interact with CD4+ tumor-infiltrating lymphocytes and predict adverse patient outcome in human laryngeal squamous cell carcinoma.

BACKGROUND: Tumor-associated macrophages (TAMs) are major component in the tumor microenvironment (TME) and play regulatory role in tumor progression. We aimed to investigate the infiltration and prognostic value of TAMs in laryngeal squamous cell carcinoma (LSCC) and to reveal the underlying mechanism of TAM subgroups in tumorigenesis. METHODS: Hematoxylin and eosin (HE) staining were performed to define the tumor nest and stroma of LSCC tissue microarrays. CD206 + /CD163 + and iNOS + TAM infiltrating profiles were obtained and analyzed through double-labeling immunofluorescence and immunohistochemical staining. The recurrence-free (RFS) and overall survival (OS) curves based on the infiltration of TAMs were plotted using the Kaplan-Meier method. Infiltration of macrophages, T lymphocytes and their corresponding subgroups were analyzed in fresh LSCC tissue samples by flow cytometry. RESULTS: We found that CD206+ rather than CD163+ M2-like TAMs were the most enriched population in the TME of human LSCC. CD206+ macrophages localized mostly in the tumor stroma (TS) rather than the tumor nest (TN) region. In contrast, relatively low infiltration of iNOS+ M1-like TAMs were found in the TS and almost none in the TN region. High level of TS CD206+ TAM infiltration correlated with poor prognosis. Interestingly, we identified a HLA-DRhigh CD206+ macrophage subgroup that was significantly associated with the tumor-infiltrating CD4+ T lymphocytes and showed different surface costimulatory molecule expression than that of the HLA-DRlow/-CD206+ subgroup. Taken together, our results indicate that HLA-DRhigh-CD206+ is a highly activated subgroup of CD206 + TAMs that may interact with CD4 + T cells through MHC-II axis and promote tumorigenesis.

Predictive value of pathological carcinoma size in patients with T2 glottic laryngeal squamous cell carcinoma.

BACKGROUND: T2N0M0 glottic laryngeal squamous cell carcinoma (LSCC) is a common type of laryngeal cancer. OBJECTIVES: The objective of this research was to assess the predictive value of tumor size for the rates of overall survival (OS) and disease-free survival (DFS) as determined by postoperative pathological examination in patients with T2 LSCC. METHODS: A retrospective study was conducted on 535 successive patients with T2 glottic LSCC who underwent operation from 2005 to 2010. The effect of tumor size on OS and DFS results was evaluated by the affected area. RESULTS: Of the cohort, 528 (98.7%) were male, and 7 (1.3%) were female, with an average age of 60.1 ± 9.4 years. The 10-year DFS and OS rates were 72.1% and 76.3%, respectively. The tumor diameter and area cut-off values that best discriminated OS and DFS rates were 1.35 cm and 1 cm2, respectively. Patients with glottis carcinoma with a longer tumor diameter and larger tumor area had inferior OS and DFS rates. Tumor diameter and tumor area were independent predictive factors for the rates of OS and DFS in patients with T2 glottic LSCC. CONCLUSION AND SIGNIFICANCE: This research showed that patients with T2 glottic LSCC with a carcinoma diameter >1.35 cm or a tumor area >1 cm2 have worse survival outcomes. These factors independently predict survival outcomes in patients.

Time-varying association of second primary malignancy and long-term survival outcomes in patients with head and neck cancer.

A high risk of developing second primary malignancy (SPM) has been reported among head and neck cancer patients. Here, we aimed to statistically quantify the impact of SPM development on the survival of head and neck cancer patients. Our study was conducted using the Surveillance, Epidemiology and End Results database to collect the data of 48 316 patients who received curative surgical resection for initial primary head and neck squamous cell carcinoma (IP-HNSCC) in 1975 to 2019. SPM diagnosis was treated as a time-varying covariate and multivariable Cox regression analysis was conducted to estimate the association between SPM development and survival, overall or by the subsite of IP-HNSCC. Of the included patients, 11 238 patients (23.3%) developed SPM during the follow-up period. A significant reduction in survival was observed among patients with SPM (hazard ratio [HR] for overall survival, 3.30; 95% confidence interval [CI]: 3.20-3.41). The impact of SPM development on reduced survival was more significant in patients with localized IP-HNSCC vs regional IP-HNSCC (HROS , 3.41; 95% CI: 3.24-3.6 vs HROS , 3.18; 95% CI: 3.05-3.31; P for interaction <.001). The survival impact of SPM development was more evident in younger patients than in older patients. SPM in lung and bronchus was associated with the most pronounced reduction in survival, overall and across all subsites of HNSCC. Our results indicated that SPM development led to a significant reduction in survival. A greater survival benefit may be achieved through intensive surveillance for SPM in lung and bronchus targeting younger patients and those with localized HNSCC.

Trends in the surgical management of parapharyngeal space tumors: A single-center retrospective analysis.

PURPOSE: Surgery remains the mainstay treatment for parapharyngeal space (PPS) tumors. Given the rapid advance and increasing usage of endoscopic and robotic techniques, we aimed to investigate the surgical trends of PPS tumors in our institution and analyze their impact on patients' treatment outcomes. MATERIALS AND METHODS: All patients who underwent surgical resection of PPS tumors from 2014 to 2021 at the Eye, Ear, Nose, and Throat Hospital of Fudan University were retrospectively reviewed. Student's t-test, Chi-square test, and multinomial logistic regression analyses were used to compare the surgical outcomes between groups. RESULT: Of the included 389 patients, the recipients of endoscopic surgery have largely increased in our center, with 17 of 134 cases (12.7%) in the group 2014-2017 and 187 of 255 cases (73.3%) in the group 2018-2021. The use of transoral and trans-nasal approaches increased in recent years (5.2% in 2014-2017 vs. 26.0% in 2018-2021), while that of trans-mandibular and lateral skull base approaches decreased (5.9% in 2014-2017 vs. 0.8% in 2018-2021). Decreased blood loss of operation and decreased risks of postoperative neurovascular complications were observed in the group 2018-2021. Similar findings were observed among patients receiving endoscopic surgery when compared with those receiving conventional surgery. CONCLUSION: In our institution, the overall trends in the surgical management of PPS tumors moved towards minimally invasive approaches with the assistance of endoscopy or surgical robots. The two surgical techniques were feasible and safe, and to a great extent, contributed to the improved surgical outcomes we observed in recent years.

Survival outcomes in patients with T3-4aN0M0 glottic laryngeal squamous cell carcinoma and evaluation of postoperative radiotherapy.

This study aimed to evaluate the clinical outcomes of patients with T3-4aN0M0 glottic laryngeal squamous cell carcinoma (LSCC) treated with laryngectomy, and to assess the postoperative radiotherapy (PORT) results in terms of the survival of T3-T4aN0M0 patients with negative margins. This was a retrospective review of 369 T3-4aN0M0 glottic LSCC cases. The 5-year cancer-specific survival (CSS) and overall survival (OS) rates were 67.5 and 66.7%, respectively. Patients who received total laryngectomy had worse survival [5-year CSS, 62.5%; disease-free survival (DFS), 56.2%] than those who underwent partial laryngectomy (5-year CSS, 79.3%; DFS, 65.4%). More advanced-stage cancer is a predictor of poor survival. There was no significant difference in CSS or DFS between patients with positive margins following rescue therapy and those with negative margins. Furthermore, no difference in the survival rates was observed between patients with negative margins who received PORT and those who did not (5-year DFS: 59.1 vs. 63.8%, P=0.057 and CSS: 62.5 vs. 69.5%, P=0.074). For T3-4aN0M0 glottic LSCC patients, surgical treatment remained a good option, as it can achieve satisfactory oncological outcomes. However, PORT did not increase survival in surgically managed pT3-4aN0M0 LSCC patients with negative margins.

Association between postoperative radiotherapy for young-onset head and neck cancer and long-term risk of second primary malignancy: a population-based study.

BACKGROUND: Second primary malignancy (SPM) represents the leading long-term cause of death among patients with index head and neck squamous cell carcinoma (HNSCC). We aimed to quantify the association between postoperative radiotherapy (PORT) and the risk of SPM development for index HNSCC among adolescent and young patients, who are particularly vulnerable to radiation-associated impacts due to their increased tissue susceptibilities and longer life expectancies. METHODS: This study was conducted using the Surveillance, Epidemiology, and End Results (SEER) database to collect the data of 5 year survivors of index young-onset HNSCC from 1975 to 2011. The outcome of interest was SPM, a new, metachronous malignancy after the index HNSCC. Standardized incidence ratios (SIRs) and excess absolute risks (EARs) were used to quantify the PORT-associated risks externally, and relative risks (RRs) were estimated by the multivariate Poisson regression analysis to quantify the PORT-associated risks internally. RESULTS: Of the included 2771 5 year survivors with index young-onset HNSCCs, the receipt of PORT (37.6%) was associated with higher risk of SPMs (RR, 1.23; 95% CI 1.07 to 1.43). PORT-associated risks were elevated for the majority of sites, including head and neck (RR, 1.19; 95% CI 0.95 to 1.50) and lung (RR, 1.67; 95% CI 1.18 to 2.34). With regarding to the subsites of head and neck, RRs were above unity in oral cavity squamous cell carcinoma (SCC) (RR, 1.68; 95% CI 1.39 to 2.03) and laryngeal SCC (RR, 1.02; 95% CI 0.73 to 1.43). A relatively greater RR was observed for patients younger than 35 years (RR, 1.44, 95% CI 0.37 to 5.57) and those diagnosed with localized diseases (RR, 1.16, 95% CI 0.9 to 1.5). PORT-associated risks were increased remarkably after 15 years of follow-up (RR, 1.24; 95% CI 0.97 to 1.58). CONCLUSIONS: An association was discovered between PORT treatment and increased long-term risk of SPM among patients with index young-onset HNSCC. The findings suggest long-term follow-up surveillance for these patients, particularly those with oral cavity SCC or laryngeal SCC.

Recurrence risk stratification and treatment strategies of patients with stage IVa-b hypopharyngeal squamous cell carcinoma.

BACKGROUND: Optimal treatment strategies for patients with stage IVa-b hypopharyngeal squamous cell carcinoma (HSCC) remain controversial. This study aimed to examine the high-risk factors of postoperative tumor recurrence after surgical resection of HSCC and devise individualized postoperative adjuvant treatment (POAT). METHODS: Overall, 218 patients with stage IVa-b HSCC who received surgery as initial treatment and with negative surgical margins were evaluated. Independent risk factors of recurrence were identified, and survival outcomes were compared according to recurrence risk and POAT use. RESULTS: POAT significantly improved recurrence-free survival (RFS) and overall survival (OS) only in the high-risk patients (p = 0.003 and 0.018, respectively). Compared with postoperative radiotherapy alone, postoperative chemoradiotherapy (pCRT) achieved significantly better RFS (p = 0.035) and OS (p = 0.048). CONCLUSIONS: POATs are recommended for high-risk patients with stage Iva-b HSCC, with pCRT achieving superior outcomes. Regular re-examination after tumor resection is sufficient for low-risk patients.

PregGAN: A prognosis prediction model for breast cancer based on conditional generative adversarial networks.

BACKGROUND AND OBJECTIVE: Generative adversarial network (GAN) is able to learn from a set of training data and generate new data with the same characteristics as the training data. Based on the characteristics of GAN, this paper developed its capability as a tool of disease prognosis prediction, and proposed a prognostic model PregGAN based on conditional generative adversarial network (CGAN). METHODS: The idea of PregGAN is to generate the prognosis prediction results based on the clinical data of patients. PregGAN added the clinical data as conditions to the training process. Conditions were used as the input to the generator along with noises. The generator synthesized new samples using the noises vectors and the conditions. In order to solve the mode collapse problem during PregGAN training, Wasserstein distance and gradient penalty strategy were used to make the training process more stable. RESULTS: In the prognosis prediction experiments using the METABRIC breast cancer dataset, PregGAN achieved good results, with the average accurate (ACC) of 90.6% and the average AUC (area under curve) of 0.946. CONCLUSIONS: Experimental results show that PregGAN is a reliable prognosis predictive model for breast cancer. Due to the strong ability of probability distribution learning, PregGAN can also be used for the prognosis prediction of other diseases.

Management of clinically node-negative glottic squamous cell carcinoma patients according to risk-scoring model for occult lymph node metastases.

Background: Glottic squamous cell carcinoma (GSCC) is the most prevalent type of laryngeal carcinoma. The value of prophylactic lymph node dissection (LND) in resected GSCC remains controversial. This study aims to quantitatively assess the probability of occult lymph node metastasis (LNM) for GSCC patients and devise individualized postoperative radiotherapy strategies. Methods: A total of 1319 patients with GSCC were retrospectively analyzed. Results: GSCC patients with T1-T2 stages showed significantly lower LNM rate than those with T3-T4 stages. For patients with T3-T4 GSCC, multivariate logistic analyses indicated that three factors-maximum tumor diameter (MTD) of more than 2.0 cm, relatively low differentiation, and tumor invasive depth of no less than 1.0 cm-were independent risk factors for the existence of LNM. A predictive nomogram was established based on these factors. The accuracy and validity of our model were verified by 0.716 and remained at 0.717 after 1000 bootstrapping. The calibration curve was also plotted and showed a favorable agreement. The patients were stratified into two groups based on their individual LNM risk points. Possible LNM rates for low-risk and high-risk subgroups were 4.7% and 25.2%, respectively. Conclusions: A new post-operative strategy selection flow chart was established based on our newly created nomogram which can effectively predict the individualized possibility of occult LNM for GSCC patients. For clinical T3-4N0 patients in the high-risk subgroup, prophylactic dose post-operative radiation therapy is recommended. However, for all those clinically diagnosed as T1-2N0 stage, regular follow-up is sufficient in view of the low occult LNM rate.Level of Evidence: 2a.

Tumor-Infiltrating PD-L1+ Neutrophils Induced by GM-CSF Suppress T Cell Function in Laryngeal Squamous Cell Carcinoma and Predict Unfavorable Prognosis.

PURPOSE: Chronic inflammation contributes to tumor initiation, progression, and immune escape. Neutrophils are the major component of inflammatory response and participate in the tumorigenesis process. However, compared to other immune cells in the tumor microenvironment of laryngeal squamous cell carcinoma (LSCC), neutrophils, especially the tumor-associated neutrophils (TANs), have not yet been comprehensively explored. The mechanism for regulating the crosstalk between TANs and tumor cells still remains unclear. MATERIALS AND METHODS: The distribution profiles and phenotypic features of neutrophils and other inflammatory immune cell populations from a large LSCC patient cohort were systemically analyzed. Co-culturing of peripheral blood associated neutrophils (PANs) and TANs with PBMCs was performed, and the immunosuppression effect on T-cells was examined. RESULTS: LSCC microenvironment is highly inflammatory with remarkable TANs infiltration, which is often associated with unfavorable prognosis and advanced clinical stage. We find that TANs in LSCC display morphologically immature and lower apoptosis, exhibit distinctively immunosuppressive phenotype of high PD-L1, and suppress CD8+ T lymphocytes proliferation and activation. We subsequently discover that PD-L1+TANs induced by LSCC-derived GM-CSF potently impair CD8+ T-cells proliferation and cytokines production function, which are partially blocked by a PD-L1-neutralizing antibody. Clinical data further support GM-CSF as an unfavorable prognostic biomarker and reveal a potential association with inflammatory immune cell infiltration, in particular neutrophils. CONCLUSION: Tumor-infiltrating PD-L1+ neutrophils induced by LSCC-derived GM-CSF suppress T cell proliferation and activation in the inflammatory microenvironment of LSCC and predict unfavorable prognosis. These TANs cripple antitumor T cell immunity and promote tumor progression. Our findings provide a basis for targeting PD-L1+TANs or GM-CSF as a new immunotherapeutic strategy for LSCC.

Risk stratification and corresponding postoperative treatment strategies for occult contralateral lymph node metastasis in pyriform sinus squamous cell carcinoma patients with ipsilateral node-positive necks.

BACKGROUND: To quantitatively predict the probability of occult contralateral lymph node metastasis (cLNM) for pyriform sinus squamous cell carcinoma (PSSC) patients with ipsilateral node-positive necks to guide postoperative adjuvant treatment. METHODS: Two hundred and twenty-seven PSSC patients with ipsilateral lymph node metastasis (iLNM) were retrospectively analyzed. RESULTS: Multivariate logistic analyses showed that five factors including maximum tumor diameter (MTD) of more than 4.0 cm, existence of tumor extension across the midline (EAM), internal jugular vein adhesion (IJVA), lymphovascular invasion (LVI), and lymph nodal fusion (LNF) were independent risk factors for cLNM. A predictive nomogram was created based on these factors. The accuracy and validity of our model were verified by concordance index (C-index) 0.862 [95% confidence interval (CI): 0.810-0.914] in development cohort and 0.860 (95% CI: 0.820-0.900) after 1,000 bootstrapping. The calibration curve also showed a relatively favorable agreement. We then stratified patients into three groups based on their cLNM risk scores. Possible cLNM rates for low-risk, moderate-risk, and relatively high-risk subgroups were 3.6%, 21.8%, and 60.7%, respectively. CONCLUSIONS: A new postoperative adjuvant radiotherapy (PART) strategy selection flow chart was created for PSSC patients based on our newly built nomogram which can effectively predict the individualized possibility of cLNM. For patients in high-risk subgroup, therapeutic-dose PART is highly recommended even for those with contralateral clinical N0 neck disease. For those in moderate-risk subgroup, prophylactic-dose PART is recommended. However, for patients in low-risk subgroup, regular follow-up is sufficient given the extremely low occult cLNM rate.