Neu_CCL3 enhances responsiveness to anti–PD-1 therapy in syngeneic MOC1 and MOC2 tumor models. A, Schematic of the experimental design and treatment schedule. Neutrophil depletion was performed using anti-Ly6G (clone 1A8; 200 μg i.p. every 3 days for five doses) and was validated by flow cytometry. B and C, MOC2 model: (B) representative images of excised subcutaneous tumors at endpoint; C, tumor growth curves (n = 5 mice per group), shown as mean ± standard deviation (SD). D and E, MOC1 model: (D) representative images of excised subcutaneous tumors at endpoint; E, tumor growth curves (n = 5 mice per group), shown as mean ± SD. F and G, MOC2 model: (F) representative flow cytometry plots of intratumoral CD8+ T cells (gated on live CD45+CD3+ cells) across the indicated treatment groups and (G) quantification of intratumoral CD8+ T-cell frequencies. H and I, MOC1 model: (H) representative flow cytometry plots of intratumoral CD8+ T cells (gated on live CD45+CD3+ cells) across the indicated treatment groups and (I) quantification of intratumoral CD8+ T-cell frequencies. Statistical analyses were performed using a two-sided Student t test. ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001. Flow cytometry plots are representative of one mouse per group; quantification includes all mice.

Malignant epithelial cells, B cells, and macrophages in Rs promote neutrophil recruitment. A, UMAP visualization of all epithelial cells, colored by cluster identity. Cells within the dashed line were identified as MT based on copy-number variation analysis. B, Single-cell CNV heatmap, with each row representing one epithelial cell and each column representing a chromosomal region. C, Pathways enriched among DEGs in MT cells from R vs. NR groups (GSEA). D, Box plot quantifying the expression levels of key chemokines (R, n = 6; NR, n = 7). *, P < 0.05; **, P < 0.01. E, UMAP of B cells, colored by subtype identity. F, Pathways enriched among DEGs in B cells from R vs. NR groups (GSEA). G, Number of key ligand–receptor pairs from B-cell subsets to neutrophils in different response groups. H, CellChat bubble heatmap showing predicted interactions between BM_FCRL4 B cells (ligand) and neutrophils (receptor) in R vs. NR groups. I, UMAP of myeloid cells, colored by subset identity. J, Box plot comparing the proportions of each myeloid cell subset between R and NR patients. K, Bar plot of GSEA results for pathways enriched in DEGs between total macrophages from R and NR patients. L, CellChat bubble heatmap showing predicted interactions between macrophages (ligand) and neutrophils (receptor) in R vs. NR groups.

PURPOSE:Hypopharyngeal squamous cell carcinoma (HPSCC) has a poor prognosis. Although neoadjuvant chemoimmunotherapy (nCIT) is promising, responses are heterogeneous, and the PD-L1 combined positive score (CPS) inadequately stratifies benefit. We sought biomarkers to guide patient selection. PATIENTS AND METHODS:In this prospective, single-center, single-arm phase II trial, patients with resectable locally advanced HPSCC received two cycles of neoadjuvant toripalimab, albumin-bound paclitaxel, and nedaplatin. The primary endpoint was the pathologic complete response (pCR) rate. Pretreatment tumor biopsies from a subset of patients (n = 13) were analyzed by single-cell RNA sequencing to identify determinants of response. Findings were validated in a larger cohort (n = 60) using bulk RNA-seq and immunohistochemistry. RESULTS:Among 70 evaluable patients, the objective response rate was 82.7%. Of the 64 patients who underwent surgery, the pCR rate was 29.7% (95% confidence interval, 18.9%-42.7%). Baseline PD-L1 CPS was not associated with pathologic response (P = 0.313). Single-cell analysis revealed that the pretreatment tumor microenvironment of responders was significantly enriched with a proinflammatory neutrophil subset characterized by high expression of CCL3 (Neu_CCL3). A gene signature score derived from this subset was a strong and independent predictor of pCR (AUC = 0.788), significantly outperforming PD-L1 CPS (AUC = 0.621). CONCLUSIONS:The efficacy of nCIT in HPSCC is predetermined by a baseline immune architecture orchestrated by a CCL3+ neutrophil subset. The Neu_CCL3 gene signature is a promising, clinically translatable biomarker that can fill a critical gap in precision immunotherapy for HPSCC.

An early-activated CD8_CD69 T cell subset is associated with response to nCIT. A, UMAP of T and NK cells, colored by subset identity. B, UMAP showing the expression of canonical marker genes in the major T-cell phenotypes. C, Heatmap showing the expression of canonical marker genes (normalized) across each T/NK cell subset. D, Box plot comparing the proportions of each T/NK cell subset (as a percentage of total T/NK cells) between R and NR patients (R, n = 6; NR, n = 7; each dot represents one patient). E, Box plots showing the cytotoxicity (left) and exhaustion (right) gene signature scores for each CD8+ T-cell subset (signature genes are detailed in “Patients and Methods”). F, Bubble chart showing enriched pathways among DEGs for each CD8+ T-cell subset (GSEA; only significantly enriched pathways are shown). G, CellChat bubble heatmap showing significant ligand–receptor interactions between MT cells and CD8_CD69 T cells. H, Bar graph showing the Spearman correlation coefficients between the infiltration fraction of each T/NK cell subset and the percentage of tumor shrinkage after neoadjuvant therapy.

Treatment-related adverse events by grade (N=70)

The Neu_CCL3 subset is enriched in Rs, and its signature score robustly predicts immunotherapy outcomes. A, UMAP of neutrophils, colored by subset identity. B, Box plot comparing the proportions of each neutrophil subset between R and NR patients. C, Bubble chart showing pathways enriched among DEGs for each neutrophil subset (GSEA; only significantly enriched pathways are shown). D, Heatmap showing the normalized expression of canonical marker genes for each neutrophil subset. E, Schematic diagram of the HPSCC validation cohort receiving nCIT. F and G, Representative dual-marker multiplex IF (mIF) images of pretreatment tumors from R and NR patients in mIF cohort 1 (n = 56; scale bar, 200 μm; F) and the corresponding quantification (G). Each point in the box plot represents one sample. ****, P < 0.0001. H, Neu_CCL3 signature scores in validation cohort 2 (n = 60), compared between R and NR patients. I, Area under the ROC curve (AUC) for pCR prediction by the Neu_CCL3 signature and PD-L1 CPS score in validation cohort 2. J, Forest plot of a multivariate logistic regression analysis in validation cohort 2 showing the association of clinical factors and the Neu_CCL3 signature with pCR.

Single-cell transcriptomic landscape of the HPSCC TME before nCIT. A, Schematic diagram of the study design. B, Pre- and posttreatment CT scans and H&E-stained tissue sections from one R and one NR. Scale bar, 400 μm. C, UMAP visualization of 97,099 high-quality cells from 13 pretreatment HPSCC tumors, colored by major cell lineage. D, Box plot comparing the relative abundances of major cell lineages in the TME between Rs (n = 6) and NRs (n = 7). The middle line indicates the median, and the box denotes the interquartile range (IQR). E, Neutrophil infiltration in the IHC validation TMA cohort (n = 56), with a box plot quantifying the percentage of positive cells. F, Macrophage infiltration in the IHC validation TMA cohort (n = 56), with a box plot quantifying the percentage of positive cells. Two-sided Wilcoxon rank-sum test, ***P < 0.001; ns, no significant difference. Scale bar, 200 μm. G, GSEA of hallmark pathways.

OBJECTIVE:Pharyngolaryngeal venous malformations (VMs) can cause airway obstruction and bleeding. This study evaluates the effectiveness of suture ligation combined with sclerotherapy for VM management. METHODS:We retrospectively analyzed 53 patients with pharyngolaryngeal VMs treated between January 2017 and September 2024. Preoperative evaluation included fibreoptic laryngoscopy and imaging. Suture ligation aimed to transfix and obstruct the VM, followed by intralesional bleomycin injection. RESULTS:The study included 23 males and 30 females (mean age 50.19 years). Most cases (71.7%) were solitary lesions. A single procedure achieved complete VM resolution in 79.25% of patients, with only one case showing minimal (less than 50%) reduction. Reoperation achieved complete regression in an additional 16.98% (nine cases). No prophylactic or emergent tracheostomy was necessary. One patient had a pre-existing tracheostomy. No serious postoperative complications occurred. CONCLUSION:Suture ligation combined with sclerotherapy offers a safe, minimally invasive, and effective approach for treating pharyngolaryngeal VMs. This technique achieves high success rates with minimal complications, promoting patient satisfaction. LEVEL OF EVIDENCE: 4:

6116 Background: Hypopharyngeal squamous cell carcinoma (HPSCC) has a poor prognosis. Although neoadjuvant chemoimmunotherapy (nCIT) is promising, responses are heterogeneous and PD-L1 combined positive score (CPS) inadequately stratifies benefit. We sought biomarkers to guide patient selection. Methods: In this prospective, single-center, single-arm phase II trial, patients with resectable locally advanced HPSCC received two cycles of neoadjuvant toripalimab, albumin-bound paclitaxel, and nedaplatin. The primary endpoint was the pathological complete response (pCR) rate. Pre-treatment tumor biopsies from a subset of patients (n=13) were analyzed by single-cell RNA sequencing (scRNA-seq) to identify determinants of response. Findings were validated in a larger cohort (n=60) using bulk RNA sequencing and immunohistochemistry. Results: Among 70 evaluable patients, the objective response rate was 82.7%. Of the 64 patients who underwent surgery, the pCR rate was 29.7% (95% CI, 18.9%–42.7%). Baseline PD-L1 CPS was not associated with pathological response (P=0.313). Single-cell analysis revealed that the pre-treatment tumor microenvironment of responders was significantly enriched with a pro-inflammatory neutrophil subset characterized by high expression of CCL3 (Neu_CCL3). A gene signature score derived from this subset was a strong and independent predictor of pCR (AUC = 0.788), significantly outperforming PD-L1 CPS (AUC = 0.621). Conclusions: The efficacy of nCIT in HPSCC is predetermined by a baseline immune architecture orchestrated by a CCL3+ neutrophil subset. The Neu_CCL3 gene signature is a promising, clinically translatable biomarker that can fill a critical gap in precision immunotherapy for HPSCC. Clinical trial information: ChiCTR2400081826.

Study design and clinical efficacy of neoadjuvant toripalimab plus chemotherapy in HPSCC. A, CONSORT flow diagram illustrating patient recruitment, treatment allocation, and analysis populations. A total of 70 eligible patients were included in the intention-to-treat analysis, and 64 patients underwent surgery. B, Waterfall plot showing the best percentage change in maximum tumor diameter from baseline for each patient (n = 70). Colors indicate radiographic response according to Response Evaluation Criteria in Solid Tumors, version 1.1. C, Sankey diagram illustrating the concordance between radiographic response categories [CR, partial response (PR), stable disease/PD] and pathologic response categories (MPR vs. non-MPR) in the surgical cohort (n = 64). D, Contingency table analysis shows that patients who achieved an objective radiographic response (defined as CR or PR) were significantly more likely to reach MPR (two-sided Fisher’s exact test; n = 64 surgically treated patients). E, Sankey diagram showing the relationship between pretreatment PD-L1 CPS status (≥1 vs. <1) and pathologic response (n = 64). F, Comparison of MPR rates stratified by PD-L1 CPS status. The P value was calculated using a two-sided Fisher’s exact test (n = 64). G, Swimmer plot depicting the clinical course for each patient (n = 70), including the duration of neoadjuvant therapy, timing of surgery, and follow-up status (recurrence or death). H, Kaplan–Meier OS curve for all treated patients (n = 70). I, Kaplan–Meier EFS curve for all treated patients.

Interactions between the Neu_CCL3 subset and T-cell or macrophage subsets. A, Number of significant ligand–receptor pairs from Neu_CCL3 cells to T-cell subsets in different response groups. B, Number of significant ligand–receptor pairs from Neu_CCL3 cells to macrophage subsets in different response groups. C, Bubble heatmap showing ligand–receptor interactions between Neu_CCL3 cells and CD8+ T cells and macrophages. D, Heatmap showing the activity (left) and regulatory potential (right) of top ligands in Neu_CCL3 cells that drive the antitumor potential of CD8+ T cells and macrophages. E, Bubble heatmap showing ligand–receptor interactions between macrophages and CD8+ T cells. F, Summary diagram of the pre-nCIT TME dynamics in patients with HPSCC who achieved pCR, by Figdraw.

Background:The fat mass and obesity-associated protein (FTO) is implicated in various diseases and acts as a demethylase for the most abundant modification of mRNA, namely N6-methyladenosine (m6A) modification. It is known that FTO may play an oncogenic role or a tumor-suppressor role in different malignancies. The aim of this study was to investigate the functional roles of FTO in regulating biological processes related to hypopharyngeal squamous cell carcinoma (HSCC). Methods:Using immunohistochemistry, quantitative real-time polymerase chain reaction (RT-qPCR), and Western blot analysis, we compared the expression levels of FTO in HSCC tissues to adjacent non-cancerous tissues. Furthermore, we evaluated the prognosis of patients with hypopharyngeal cancer in relation to FTO expression levels. In vitro, the Cell Counting Kit-8 (CCK8), wound healing assay, migration and invasion assays were used to identify roles of FTO in HSCC cells FaDu. Tumor xenografts in nude mice were used to disclose the effect of FTO in vivo. Then, transcriptome RNA sequencing (RNA-seq) assays were applied to screen for possible target genes. To confirm the specific site for modulating the expression of the target gene, we used the SRAMP database and methylated RNA immunoprecipitation PCR (MeRIP-PCR). Results:The results showed that FTO was highly expressed in hypopharyngeal cancer tissues and was correlated with clinicopathology of patients. FTO promoted the proliferation, invasion and migration of hypopharyngeal cancer cells in vitro through its demethylase action. In vivo experiments showed that FTO promoted the growth of subcutaneously implanted tumors of hypopharyngeal cancer cells and their metastasis. Moreover, we revealed that FTO affected the malignant biological behavior of hypopharyngeal cancer cells by regulating the m6A modification level of SERPINE1 mRNA. FTO promoted epithelial-mesenchymal transformation (EMT) of hypopharyngeal cancer cells through the SERPINE1 signaling axis. Conclusions:Our study highlighted the functional significance of the FTO/SERPINE1 axis in tumorigenesis of HSCC. Targeting FTO holds promise as a new therapeutic strategy for HSCC.

Background and Objectives: The hematopoietically expressed homeobox protein (HHEX) is closely related to the occurrence and development of a variety of neoplasms. The aim of this study was to explore the role of HHEX expression in nasopharyngeal carcinoma (NPC) tissue and its clinical significance. Methods: In this retrospective cohort study, we examined the expression of HHEX, the sensitivity of 8 common chemotherapeutic agents, in 76 NPC tissue specimens. The relationship between HHEX expression and clinical pathological factors, chemotherapy sensitivity and prognosis were analyzed by t test, chi-square test, analysis of variance, rank sum test, Spearman correlation analysis, log-rank test and multivariable Cox proportional hazard models. Results: HHEX expression was significantly upregulated in NPC tissues compared with chronic inflammatory tissues (P<0.001). HHEX expression was negatively correlated with T stage (r=-0.55, P<0.001), N stage (r=-0.36, P=0.006), M stage (r=-0.27, P=0.022) and clinical stage (r=-0.63, P<0.001). Chemotherapy resistance to paclitaxel was significantly lower in NPC tissue with high HHEX expression than that with low HHEX expression (P=0.004). The periods of progression-free survival in patients with low HHEX expression were significantly shorter than those in patients with high HHEX expression (P=0.041). Conclusion: HHEX was highly expressed in NPC tissue and may be involved in the NPC occurrence and development process. The levels of HHEX expression may possess significant prognostic value for individuals suffering from NPC. Monitoring of HHEX expression in these patients could provide critical insights into their sensitivity to chemotherapeutic agents and contribute to the evaluation of overall treatment outcomes.

Glioma is one of the most aggressive and lethal brain tumors, with poor prognosis and limited treatment options. This study examined the function of CLEC18B in glioma development and its viability as a predictive biomarker. Pan-cancer research demonstrated that CLEC18B is dysregulated in several tumor types, with elevated expression associated with reduced overall survival (OS) and disease-specific survival (DSS) in patients with different malignancies, including glioma. CLEC18B was markedly increased in glioblastoma (GBM) and lower-grade glioma (LGG) tissues relative to normal tissues, and its elevated expression correlated with worse survival outcomes in both LGG and GBM patients. CLEC18B expression was an independent predictive indicator for OS and DSS in GBM, with expression levels being affected by DNA methylation status. We investigated the regulatory mechanisms governing CLEC18B expression and found SP1 as a major transcription factor that directly modulates CLEC18B. Our findings validated that SP1 associates with the CLEC18B promoter, and the silencing of SP1 resulted in a substantial decrease in CLEC18B expression. The suppression of CLEC18B functionally decreased glioma cell proliferation, motility, and invasion in vitro, and lowered tumor development in vivo. Furthermore, CLEC18B knockdown modified the Wnt/β-catenin/EMT signaling pathway by decreasing mesenchymal markers and increasing epithelial markers. Administration of a Wnt/β-catenin agonist partially mitigated the consequences of CLEC18B knockdown, indicating that CLEC18B facilitates glioma growth via the stimulation of this pathway. In conclusion, CLEC18B is crucial to glioma development, serving as a principal regulator of cell proliferation, migration, and invasion via the Wnt/β-catenin/EMT pathway. CLEC18B may function as a prospective prognostic biomarker and therapeutic target for glioma therapy.

Osteoarthritis (OA) is a degenerative joint disease characterized by progressive articular cartilage loss, osteophyte formation, and synovial inflammation. For OA, most of the Western medical treatments are surgery, drugs, etc. However, the effects of these methods are limited, and there are many side effects. Studies have confirmed that astragaloside (AST) can reduce the pathological changes of OA, but the molecular mechanism is still unclear. Therefore, we investigated the mechanism of AST in the treatment of OA based on network pharmacology, molecular docking simulation, and animal experiments. First, search the related target of AST and OA in the database. Then the target genes of AST and OA were intersected to construct the interaction between target protein–protein interaction (PPI) network. Through software analysis, PPI core subnetwork modules and top 10 key targets were obtained and enriched for analysis. Molecular docking simulation was used to predict the binding pattern and binding ability between AST and key target proteins. AST and saline were injected into the knee cavity of osteoarthritis rabbits, respectively, labeled as AST group and OA group. After 4 weeks, the therapeutic effect of OA in the two groups was compared by histological staining, MRI, and qRT-PCR to detect and verify the expression of key target genes. The top 10 key intersection target genes were obtained. GO enrichment analysis obtained 625 annotation items, KEGG enrichment analysis obtained 111 signal pathways. Molecular docking simulation showed that AST and key target proteins showed good binding force. Animal experiments displayed that AST had obvious therapeutic effect on OA; knee fluid accumulation and bone marrow injury were significantly reduced. qRT-PCR results showed that, compared with OA group, the expressions of SRC and TLR4 in AST group were significantly increased, while the expressions of ALB and ESR1 were decreased; the expression of other genes had no significant difference. AST treatment of OA is characterized by multi-target and multi-pathway interactions, and its key target genes act on OA by affecting endothelial cell permeability, regulating synovial, chondrocyte survival, proliferation, and extracellular matrix synthesis, etc., and its molecular mechanism involves the Src/PI3K/Akt, NF-κB, MAPK, and TLRs signaling pathways.

The KRAS mutation is highly prevalent in NSCLC and is associated with poor efficacy of immunotherapy. Nevertheless, the impact of KRAS mutation, mutation subtypes, and co-mutations on the effectiveness of immunotherapy remains uncertain. This study aimed to assess the influence of the KRAS mutation on the effectiveness of immunotherapy in NSCLC, specifically examining different subtypes of KRAS mutations and co-mutations. We performed an extensive search of multiple databases, covering the period from January 1, 2000, to December 5, 2023. A total of 24 articles met our inclusion criteria and were included in this study. A comparative analysis assessed the influence of different subgroups, including KRAS mutation, KRAS wild-type, KRAS G12C mutation, KRAS G12D mutation, and KRAS with co-mutations in NSCLC with immunotherapy. The study outcomes include HR, with corresponding 95

Background: cervical squamous cell carcinoma (CSCC) is the second gynecological tumors that seriously threaten women's life quality. Circular RNA (circRNA) is related with cervical cancer carcinogenesis and radiosensitivity. Aim: To investigate the performance of hsa_circ_0007905 (circSTX6) on regulating cellular activities and radiosensitivity in CSCC. Methods: The relative expression of circSTX6 in different tissue samples was detected by RT-qPCR. The cellular activity influence of circSTX6 in cervical cancer cells was measured by CCK-8 and Transwell assays. The survival fractions of cancer cells were detected after the radiation treatment to explore the relationship between circSTX6 and radiosensitivity of cervical cancer. The downstream miRNAs were predicted and analyzed. Rescue experiments confirmed their targeting relationship. Bioinformatic analysis was performed to identify the potential targets of miR-203a-3p. Results: circSTX6 was increased and miR-203a-3p was decreased in cervical cancer tissues and radio-resistant tissues. CircSTX6 expression was related to the patient's survival rates. CircSTX6 absence decreased cervical cancer cell proliferation and invasion while enhancing the sensitivity of cervical cancer cells to radiotherapy by regulating miR-203a-3p. RAB27B may be a target of miR-203a-3p. Conclusion: circSTX6 may be a clinical prognostic biomarker in CSCC. The absence of circSTX6 inhibits cellular behaviors and increases the sensitivity of cervical cancer cells to radiation by modulating miR-203a-3p/RAB27B axis.

BACKGROUND:Identifying the key molecular targets in hypopharynx squamous cell carcinoma (HSCC) is crucial for understanding this prevalent and highly fatal type of head and neck tumor. The study aims to enhance comprehension of the HSCC process by accurately identifying these key molecular targets. MATERIALS AND METHODS:In this study, we examined 47 clinical tissue samples from individuals diagnosed with HSCC using RNA-seq high-throughput assay. Quantitative real-time PCR (RT-PCR) was used to compare long non-coding RNA (lncRNA) bladder cancer-associated transcript 1 (BLACAT1) expression in HSCC tissues versus adjacent non-tumor tissues. The influence of highly expressed lncRNA BLACAT1 on prognostic survival was assessed. Subsequently, we cultured human pharynx squamous cell carcinoma FaDu cells. After reducing lncRNA BLACAT1 expression, we assessed FaDu cell proliferation, invasion, and migration using Cell Counting kit-8 (CCK-8) assay, colony formation assay, EUD assay, Transwell assay, and scratch assay. Additionally, liquid chromatography-tandem mass spectrometry/mass spectrometry (LC-MS/MS) and western blotting analysis were used to analyze proteins that bind to lncRNA BLACAT1. During in vivo experiments, mice received subcutaneous injections of FaDu cells transfected with lncRNA BLACAT1 shRNA or Scr plasmid (Control) in the dorsal region to observe and compare tumor growth. Lastly, tumor tissues underwent hematoxylin-eosin (HE) and immunohistochemical (IHC) staining. RESULTS:lncRNA BLACAT1 was screened as one of the most significant genes among the group of differentially expressed lncRNAs. RT-PCR exhibited elevated lncRNA BLACAT1 expression in HSCC tissues when compared to non-tumor tissues (p < 0.001). Furthermore, increased lncRNA BLACAT1 expression correlated with advanced clinical stages, heightened lymphatic invasion, and a poor prognosis. Subsequent in vitro experiments solidified our observations, demonstrating lncRNA BLACAT1's promotion of HSCC cell proliferation (p < 0.05), migration (p < 0.01), and invasion (p < 0.01) compared with the control group. Moreover, LC-MS/MS identified signal transducer and activator of transcription 3 (STAT3) and Prohibitin 2 (PHB2) as lncRNA BLACAT1-binding proteins and sh-lncRNA BLACAT1 inhibits STAT3/AKT phosphorylation (p < 0.01) and alters the subcellular distribution of PHB2 and P21 compared with the control group (p < 0.01). Moreover, in vivo experiments showed that lncRNA BLACAT1 inhibition suppresses tumorigenicity in an HSCC xenograft model compared to the control group (p < 0.01). CONCLUSIONS:lncRNA BLACAT1 is highly expressed in HSCC tumor tissues and plays a crucial role in the development of HSCC in vitro and in vivo. This increased expression may be caused by STAT3/AKT pathway activation, consequently inhibiting P21 expression through PHB2.

Background Cells in the body reside in a dynamic microenvironment subjected to various physical stimuli, where mechanical stimulation plays a crucial role in regulating cellular physiological behaviors and functions. Aim of Review Investigating the mechanisms and interactions of mechanical transmission is essential for understanding the physiological and functional interplay between cells and physical stimuli. Therefore, establishing an in vitro biomechanical stimulation cell culture system holds significant importance for research related to cellular biomechanics. Key Scientific Concepts of Review In this review, we primarily focused on various biomechanically relevant cell culture systems and highlighted the advancements and prospects in their preparation processes. Firstly, we discussed the types and characteristics of biomechanics present in the microenvironment within the human body. Subsequently, we introduced the research progress, working principles, preparation processes, potential advantages, applications, and challenges of various biomechanically relevant in vitro cell culture systems. Additionally, we summarized and categorized currently commercialized biomechanically relevant cell culture systems, offering a comprehensive reference for researchers in related fields.

AbstractBackgroundSevere acute respiratory syndrome coronavirus 2 disease (COVID‐19) has caused a worldwide challenging and threatening pandemic. We aimed to assess the safety and efficacy of the COVID‐19 vaccines in Non‐Small Cell Lung Cancer (NSCLC) patients.MethodsPatient self‐reported adverse events related to vaccines were recorded by follow‐up through a uniform questionnaire. Survival analysis was performed by Kaplan–Meier method. A multivariate analysis was performed by the Cox proportional hazard regression model to determine the effect of each variable on the survival of lung cancer patients.ResultsA total of 860 patients with NSCLC on treatment were enrolled. Mean age was 57 years in patients with early stage group and 62 years in advanced stage group. The vaccination rate was 71.11% for early‐stage patients and 19.48% for advanced‐stage patients; most of them (86.5%) received the COVID‐19 inactivated virus (Vero cell) vaccine (Coronavac; Sinovac). The most common systemic adverse reaction was weakness. The main reason for vaccine refusal in those unvaccinated patients was concern about the safety of vaccination in the presence of a tumor and undergoing treatment (56.9% and 53.4%). The 1‐year disease‐free survival (DFS) rate was 100% for vaccinated and 97.4% for unvaccinated early‐stage patients. Then we compared the progression‐free survival (PFS) of vaccinated (median PFS 9.0 months) and unvaccinated (median PFS 7.0 months) advanced stage patients (p = 0.815). Advanced NSCLC patients continued to be divided into groups receiving radio‐chemotherapy, immunotherapy, and targeted therapy, with no statistical difference in PFS between the groups (p > 0.05). The median overall survival (OS) of vaccinated patients was 20.5 months, and that of unvaccinated patients was 19.0 months (p = 0.478) in advanced NSCLC patients.ConclusionsCOVID‐19 vaccination is safe for Chinese NSCLC patients actively receiving different antitumor treatments without increasing the incidence of adverse reactions, and vaccination does not affect cancer patient survival.