Context Bladder cancer management is challenged by limited therapeutic targets and heterogeneous treatment responses. Ribosomal S6 kinase 4 (RSK4) has been established as an oncogenic driver in several malignancies, although its clinical significance in bladder cancer remains undefined.Objective To evaluate RSK4 protein expression in bladder cancer specimens and assess its association with clinicopathologic features and patient outcomes.Design RSK4 expression was analyzed by immunohistochemistry in a retrospective cohort of 143 bladder cancer specimens, including 93 cases represented in a tissue microarray. Statistical analyses were performed to evaluate the associations between RSK4 expression levels, standard clinicopathological parameters, and overall survival.Results RSK4 immunoreactivity was detected in 65.7% (94/143) of tumor tissue samples but in 36.5% (23/63) of matched normal urothelial tissue samples (P < 0.0001). Elevated RSK4 expression was significantly correlated with the following established markers of disease progression: muscularis propria invasion (P < 0.001), high tumor grade (P < 0.01), advanced TNM stage (P < 0.001), lymph node metastasis (P < 0.01), and distant metastasis (P < 0.05). No significant associations were observed with patient age, sex, or tumor size. Multivariate analysis confirmed RSK4 as an independent predictor of reduced overall survival (HR = 2.34, 95% CI 1.42-3.85, P < 0.001). Subcellular studies indicate that RSK4 overexpression enhances the invasive and metastatic capabilities of bladder cancer cell lines, and vice versa.Conclusions This study elucidates the expression pattern and mechanism of action of RSK4 in bladder urothelial carcinoma, confirming that its overexpression is a key factor for predicting poor prognosis. This discovery highlights the potential value of RSK4 as a significant therapeutic target, providing a new theoretical basis for improving clinical outcomes in bladder cancer.

Differences between m6A level-derived patient subtypes.

OBJECTIVE:To develop and validate radiomics models based on gadoxetate disodium (Gd-EOB-DTPA)-enhanced MRI for preoperative prediction of proliferative small hepatocellular carcinoma (sHCC) and to evaluate their prognostic value. METHODS:A total of 331 patients with pathologically confirmed sHCC from two institutions were retrospectively enrolled and divided into training (n = 147), internal validation (n = 63), external validation (n = 59), and exploratory validation (n = 62) cohorts. Independent predictors were identified using multivariate logistic regression to construct a clinical-radiological model. Intratumoral and peritumoral volumes of interest (VOIs) were manually delineated on hepatobiliary phase (HBP) images from Gd-EOB-DTPA-enhanced MRI, followed by feature extraction and selection to develop intratumoral, peritumoral, and combined radiomics models. Model performance was evaluated using the area under the receiver operating characteristic curve (AUC), interpretability was analyzed using Shapley additive explanations (SHAP), and recurrence-free survival (RFS) was assessed with the Kaplan-Meier method. RESULTS:In the external validation cohort, the combined radiomics model achieved the best predictive performance (AUC = 0.836), outperforming both the intratumoral (AUC = 0.748, P = 0.049) and peritumoral (AUC = 0.744, P = 0.044) models, with consistent results in the exploratory validation cohort (AUC = 0.817). No significant difference was observed between the intratumoral and peritumoral models (P = 0.863). All radiomics models outperformed the clinical-radiological model (P < 0.001). SHAP analysis identified original_shape_Elongation_Intra, log-sigma-2-0-mm-3D_firstorder_Maximum_Intra, and wavelet-HLH_firstorder_Kurtosis_Intra as the most influential features. Patients with proliferative sHCC showed significantly shorter RFS than those with non-proliferative sHCC (P = 0.049), and high-risk patients identified by the combined radiomics model exhibited poorer RFS (P = 0.025). CONCLUSION:The combined radiomics model provides a promising and noninvasive approach for preoperative prediction of proliferative sHCC and enables effective postoperative risk stratification, offering potential value for individualized clinical management.

Sample information and analysis in the internal cohort.

Temozolomide (TMZ) resistance is a critical factor that affects the therapeutic efficacy in glioblastoma (GBM). Glutathione peroxidase 8 (GPX8), a ROS scavenging enzyme, is associated with poor prognosis in GBM. In this study, we comprehensively studied the role and mechanism of GPX8 in GBM resistance to TMZ. We found that GPX8 was upregulated in GBM cells, tissues, and TMZ-resistant GBM cells. In U87 TMZ-resistant cells, GPX8 knockdown significantly suppressed cell proliferation, reversed the epithelial-mesenchymal transition (EMT), and sensitized cells to TMZ. Moreover, GPX8 knockdown induced mitochondrial oxidative stress, leading to apoptosis in TMZ-resistant cells. TEA domain family member 4 (TEAD4) was upregulated in GBM cells and transactivates GPX8. GPX8 interacted with collagen triple helix repeat containing-1 (CTHRC1) and promoted its expression. Overexpression of TEAD4 or CTHRC1 reversed the suppressive effect of GPX8 knockdown on the malignant phenotypes of TMZ-resistant cells and antagonized its promotive effect on mitochondrial ROS generation and apoptosis. Furthermore, overexpression of GPX8 or CTHRC1 promoted EMT, reduced ROS levels, and lowered TMZ sensitivity in resistant cells. Crucially, the p38 MAPK/FOXO3 pathway inhibitor Ade was able to reverse these effects. GPX8 knockdown increased GBM sensitivity to TMZ, inhibited EMT, and elevated ROS levels in both xenograft models and glioma organoids. Overall, our results elucidated that TEAD4-driven GPX8 suppresses mitochondrial oxidative stress in TMZ-resistant cells through activation of the CTHRC1/p38 MAPK/FOXO3 pathway, which promotes TMZ resistance in GBM cells. These findings suggest that GPX8 may serve as a novel therapeutic target for overcoming TMZ resistance in GBM.

Purpose: To determine whether hypoperfused yet metabolically active tumor regions—termed low-angiogenic tumor (LAT) habitats—impair glymphatic clearance in the contralateral hemisphere and predict clinical outcomes in patients with high-grade glioma (HGG) undergoing standard chemoradiotherapy. Materials and Methods: This retrospective study included 151 newly diagnosed HGG patients who underwent preoperative multiparametric MRI and received standardized treatment. LAT habitats were delineated from perfusion maps, and their metabolic activity was estimated using a weighted least squares model based on multi-voxel MRS of enhancing tumor regions. Glymphatic function was assessed via the DTI-ALPS index in the anatomically unaffected hemisphere. Partial correlation, single and serial mediation models, and Cox regression were used to evaluate interdependencies among LAT perfusion, metabolism, glymphatic dysfunction, and progression-free survival (PFS), adjusting for IDH mutation status. Results: LAT rCBV, Cho/NAA, and ALPS showed strong intercorrelations (r = 0.763, 0.591, 0.409; all p < 0.001). Mediation revealed a full pathway: LAT perfusion predicted metabolism (β = 1.25, p < 0.001), which predicted glymphatic function (β = 0.06, p < 0.001); only the indirect effect was significant. Cox analysis identified Cho/NAA and IDH—but not ALPS—as independent PFS predictors. Serial mediation confirmed that only metabolism, not glymphatic dysfunction, mediated survival. A multivariable model yielded a C-index of 0.922. Conclusions: Metabolically active LAT habitats remotely impair glymphatic function and contribute to recurrence. Combined metabolic–glymphatic profiling may offer a mechanistic basis for risk stratification and therapeutic targeting in HGG.

The therapeutic efficacy of photodynamic therapy (PDT) is frequently severely compromised by the hypoxic tumor microenvironment (TME) and the robust antioxidative defense system mediated by overexpressed glutathione (GSH). While Type I PDT offers a potential solution to hypoxia, its performance remains limited by the rapid scavenging of reactive oxygen species (ROS) by intracellular GSH. To address these dual barriers, we developed a sacrificial-electron-assisted strategy based on an all-organic nanoparticle (YB NPs). This nanoparticle was co-assembled from the non-fullerene acceptor Y6 and the electron-accepting scaffold benzophenone (BP). In this design, BP functioned as sacrificial electron scavenger that efficiently drains photoinduced electrons from Y6, thereby suppressing charge recombination and facilitating the accumulation of photogenerated holes. These accumulated holes effectively oxidized intracellular GSH, disrupting the tumor antioxidant defense. Consequently, the YB NPs achieved simultaneous GSH depletion and enhanced hydroxyl radical (·OH) generation via the Type I pathway, independent of oxygen levels. In vitro and in vivo evaluations demonstrate that this synergistic mechanism significantly amplified oxidative stress and resulted in potent tumor growth inhibition. This study presents a viable approach for designing high-performance organic photosensitizers capable of modulating the TME for enhanced cancer therapy.

Summary and gene enrichment of hyper-, hypo-, NL-specific and tumor-specific methylated genes.

IntroductionTo explore the value of enhanced computed tomography (CT) -derived extracellular volume (ECV) combined with systemic immune-inflammation index (SII) in predicting tumor budding (TB) grading of rectal cancer.Materials and MethodsThe clinical and imaging data of 177 rectal cancer patients were retrospectively analyzed, and we divided them into a low-grade and medium-high group according to pathological TB count. ECV and SII values between the two groups were compared. Intra-class correlation coefficient (ICC) was used to detect the consistency of measurements among observers. Binary logistic regression was used to analyze the correlations between variables and TB grading of rectal cancer. Receiver operating characteristic (ROC) curve analysis was performed to evaluate the diagnostic efficiency of statistically significant parameters and their combination. Area under the curve (AUC), its 95% confidence interval, and the corresponding Youden index, sensitivity, and specificity were calculated.ResultsAmong the 177 rectal cancer patients, 108 were low-grade and 69 were medium-high grade. ECV values measured by two physicians showed good consistency (ICC = 0.98). ECV value of low-grade (21.76% ± 4.89%) was lower than that of medium-high grade TB group (27.91% ± 4.77%) (P < .001). SII value was lower in low-grade group (492.14 ± 239.56) than in medium-high grade TB group (825.02 ± 529.38). In the multivariate analysis, ECV value [odds ratio (OR): 1.339 (95% CI: 1.194-1.502)] and SII value [OR: 1.004 (95% CI: 1.002-1.005)] were independent risk factors for predicting TB grading. In the training set, AUCs of ECV, SII, and their combination in evaluating TB grading of rectal cancer were 0.838 (95% CI: 0.760-0.905), 0.755 (95% CI: 0.663-0.829), and 0.889 (95% CI: 0.832-0.943), respectively. In the test set, the corresponding AUCs were 0.741 (95% CI: 0.626-0.870), 0.716 (95% CI: 0.554-0.849), and 0.815 (95% CI: 0.711-0.913). Decision curve analysis (DCA) showed that the combination had higher clinical value than using ECV or SII alone.ConclusionThe combination of ECV and SII can non-invasively evaluate TB grading of rectal cancer before surgery, potentially providing a reference for preoperative risk stratification as a decision-support tool.

INTRODUCTION:This study aimed to identify perioperative risk factors for postoperative complications in patients undergoing reoperation for persistent/recurrent differentiated thyroid carcinoma (DTC). METHODS:This retrospective study included patients with persistent/recurrent DTC who underwent reoperation at the Head and Neck Surgery Department of our Hospital between June 2021 and December 2023. Postoperative complications assessed were hypoparathyroidism, lymphatic leakage, nerve injury, and postoperative bleeding. Multivariable logistic regression was used to determine factors independently associated with these complications. RESULTS:A total of 97 patients were included, with a mean age of 46.68 ± 12.82 y, and 38 of them were male. Among these patients, 59 had no postoperative complications, while 38 experienced at least one complication. Multivariable logistic analysis revealed that the number of previous surgeries (odds ratio [OR] = 3.91, 95% confidence interval [CI]: 1.04-14.74, P = 0.044), bilateral lateral cervical lymphadenectomy (OR = 4.69, 95% CI: 1.05-20.99, P = 0.043), thoracic duct ligation (OR = 4.01, 95% CI: 1.22-13.22, P = 0.022), parathyroid hormone levels at admission (OR = 0.58, 95% CI: 0.38-0.89, P = 0.013), and operative time (OR = 1.57, 95% CI: 1.01-2.44, P = 0.045) were significantly associated with postoperative complications. CONCLUSIONS:This study identified critical perioperative early warning factors for postoperative complications in patients undergoing reoperation for persistent/recurrent DTC. These findings can support healthcare professionals in the early detection, prevention, and management of complications, thereby enhancing patient outcomes and informing more effective perioperative care strategies.

Photodynamic therapy (PDT) is a promising anticancer strategy but remains limited by inefficient reactive oxygen species (ROS) generation and insufficient subcellular targeting. Here, we report a cationic engineering strategy to enhance mitochondrial targeting and ROS-amplified photodynamic tumor therapy. A neutral aggregation-induced emission (AIE) photosensitizer (PS) was stepwise converted into monocationic (PS-Bu) and dicationic (PS-PPh) derivatives, enabling precise regulation of molecular charge density. Cationic modification modulates excited-state energetics, promotes intersystem crossing, and simultaneously enhances Type I and Type II ROS generation under visible-light irradiation. The increased positive charge drives preferential mitochondrial accumulation, leading to pronounced mitochondrial membrane depolarization, intracellular ROS amplification, cell-cycle arrest, and apoptosis. Consequently, PS-PPh exhibits markedly enhanced phototoxicity toward HepG2 cells while maintaining negligible dark toxicity. In vivo, PS-PPh achieves superior tumor growth suppression in HepG2 xenograft models, accompanied by reduced proliferation, enhanced apoptosis, and alleviation of tumor hypoxia, without detectable systemic toxicity. This work establishes cationic modulation as an effective molecular design paradigm for mitochondria-targeted and ROS-enhanced photodynamic cancer therapy.

Sample information and gene list of NL-specific, tumor-specific and common methylated genes.

Characteristics of m6A peaks and methodological comparison for m6A level calculation.

Background:Kawasaki disease (KD) is a leading cause of acquired heart disease in children, with coronary artery lesions (CAL) as its most severe complication. This observational study aimed to investigate the expression of bone marrow stromal cell antigen-1 (BST1) in KD patients and its value for predicting CAL. Methods:Serum samples were collected from KD patients before intravenous immunoglobulin (IVIG) treatment and healthy controls (HC) in Xi'an Children's Hospital from July 2023 to August 2024. KD Patients were stratified into CAL and non-CAL (nCAL) groups based on echocardiography. In the discovery cohort (8 KD-CAL patients, 7 KD-nCAL patients, and 4 HC), Sera were analyzed via four-dimensional data-independent acquisition (4D-DIA) quantitative proteomics. Differentially expressed proteins (DEPs) were identified and analyzed for bioinformatics. In the validation cohort (35 KD-CAL patients, 61 KD-nCAL patients, and 30 HC), serum BST1 was validated by enzyme-linked immunosorbent assay (ELISA). Receiver operating characteristic (ROC) curve was constructed to assess the performance of BST1 in predicting CAL. Results:A total of 2575 proteins were identified by 4D-DIA proteomics, with 807 DEPs in nCAL vs HC and 883 DEPs in CAL vs HC. Compared to nCAL, 213 DEPs were identified in CAL patients, among which BST1 was significantly upregulated only in the CAL group. Functional analyses revealed enrichment in innate immune response, cell adhesion, RNA processing pathways, complement and coagulation cascades, extracellular exosomes, and cellular metabolism. ELISA confirmation showed elevated BST1 levels in CAL patients, which positively correlated with Z-scores. ROC analysis demonstrated high prognostic accuracy for CAL (AUC=0.9052). Conclusion:Serum BST1 has the potential to be a predictive biomarker for CAL in KD patients.

Live cell imaging of EML4-overexpressing H358 cells in wound healing assay.

The clinical phenotype and pathogenic mechanism of 46,XY disorders of sex development (DSD) are complex, and several pathogenic variants are identified by next-generation sequencing. However, these variants currently require additional interpretation and validation prior to their application in 46,XY DSD diagnosis and clinical guidance. Here, we identified three genetic variants in two 46,XY DSD patients by whole exome sequencing screening and Sanger sequencing validation. The pathogenicity of three genetic variants was identified by in silico analysis and functional experiments. One patient carrying the reported pathogenic variant (c.319 C > T) of NR5A1 showed a phenotype of 46,XY complete gonadal dysgenesis, which was different from the reported 46,XY partial gonadal dysgenesis. These findings suggested that the variant (c.319 C > T) of NR5A1 contributes to the clinical phenotypic heterogeneity of 46,XY DSD. The other patient carried two genetic variants, among which the c.1252 C > T variant of NR5A1 produced truncated protein and lost the transcriptional activation of NR5A1 to the targeted genes. The other c.769G > A variant of DHX37 had no significant effect on the expression level and cellular localization of DHX37, and the downstream signaling pathway of DHX37. Moreover, the in silico and structural analysis identified the c.769G > A variant of DHX37 as a deleterious variant that may affect DHX37 function. According to the American College of Medical Genetics and Genomics guidelines and relevant literature reports, combined with the patient’s clinical phenotype and pedigreed analysis, it is proposed that the likely pathogenic variant identified in this patient is the c.1252 C > T variant of NR5A1. Nonetheless, the potential pathogenicity of the DHX37 (c.769G > A) variant of this patient also merits further investigation and consideration. Our results have expanded the clinical phenotype spectrum and genetic diagnosis spectrum of 46,XY DSD, which will contribute to the accurate diagnosis and treatment guidance for 46,XY DSD patients and provide evidence-based genetic counseling for 46,XY DSD family fertility.

mRNA abundance of m6A regulators in lung tissues and genomic characteristics of G1, G2 and G3 gene groups.

Live cell imaging of control A549 cells in wound healing assay.

Live cell imaging of control H358 cells in wound healing assay.