Aim:This study evaluates the effectiveness and safety of combining radiotherapy with first-line chemotherapy and immune checkpoint inhibitors in patients with locally advanced or metastatic urothelial carcinoma (la/mUC). Methods:This retrospective study included patients with la/mUC who received first-line systemic treatment between January 2017 and December 2024. Two treatment strategies were compared: immunotherapy combined with chemotherapy followed by radiotherapy (ICRT group, n = 31) and immunotherapy combined with chemotherapy alone (ICT group, n = 73). Propensity score matching was used to reduce confounding and selection bias, yielding 30 matched patient pairs. Clinical endpoints included objective response rate (ORR), duration of response (DOR), progression-free survival (PFS), overall survival (OS), and treatment-related adverse events. Results:After propensity score matching, the ICRT group showed significantly prolonged PFS (18.70 vs. 7.93 months; P = 0.002) and OS (38.9 vs. 19.8 months; P = 0.031) compared with the ICT group. The ICRT cohort demonstrated higher PFS, OS, and ORR across all evaluated time points, with statistically significant differences at 6-month PFS, 12-month PFS, and 12-month OS (P < 0.05). Although the median DOR was numerically longer in the ICRT group (29.23 vs. 13.53 months), the difference did not reach statistical significance (P = 0.385). Radiation-related enteritis and cystitis were observed in the ICRT group, while no treatment-related deaths occurred. Multivariate Cox regression analysis identified lung metastases, elevated C-reactive protein levels, radiotherapy, and treatment response exceeding stable disease as independent predictors of PFS. Liver metastases, radiotherapy, and better treatment response were independently associated with OS. Conclusions:The addition of radiotherapy to first-line immunotherapy and chemotherapy demonstrated favorable clinical activity and an acceptable safety profile in patients with la/mUC.

Objective:While cisplatin-based chemotherapy is pivotal for advanced bladder cancer, acquired resistance remains a major obstacle. This study investigates key molecular drivers of this resistance and potential reversal strategies. Methods:We established GC (Gemcitabine and Cisplatin)-resistant T24-R and UC3-R cell lines from T24 and UM-UC-3 (UC3) cells. Transcriptomic and proteomic analyses identified differentially expressed molecules. Apoptosis and cell viability were assessed by flow cytometry and CCK-8 (Cell Counting Kit-8) assays, while RT-qPCR (Reverse Transcription Quantitative Polymerase Chain Reaction) and Western blot analyzed gene and protein expression. Immunofluorescence evaluated FAK (Focal Adhesion Kinase) phosphorylation, and a xenograft mouse model validated the findings in vivo. Results:Integrated transcriptomic and proteomic analysis identified FN1 (fibronectin) as a consistently upregulated top candidate in resistant cells (T24-R transcript log2FC = 2.8, protein log2FC = 0.9; UC3-R transcript log2FC = 3.7; all p < 0.001). Knockdown of FN1 reduced chemoresistance (Resistance Index: 5.2 in T24-R and 2.0 in UC3-R cells, p < 0.001) and enhanced apoptosis (approximately 4.5-fold in T24-R and 7.5-fold in UC3-R, p < 0.001). ITGB4 (Integrin Subunit Beta 4) was upregulated in resistant cells (transcript log2FC: 4.2 in T24-R and 3.03 in UC3-R; protein log2FC: 0.67 in T24-R; all p < 0.01). Critically, ITGB4 knockdown abolished the chemoresistance promoted by exogenous FN1, which was associated with increased FAK (Y397) phosphorylation. Conclusion:Our results demonstrate that the FN1-ITGB4 axis drives chemoresistance in bladder cancer via FAK signaling. Targeting this axis represents a promising strategy to overcome chemoresistance.

The maintenance and regulation of cancer stem cell (CSC) stemness are crucial for tumor progression; however, the mechanisms underlying tumor stemness regulation remain poorly understood. Herein, we discovered that the enhanced hypoxia-induced transforming growth factor beta induced protein (TGFBI) in bladder cancer (BLCA) promotes the establishment of a stemness loop in the tumor microenvironment, facilitating the maintenance of CSC stemness and malignant proliferation. Clinically, the upregulation of hypoxic TGFBI in BLCA correlates with malignant BLCA features and poor prognosis. Mechanically, TGFBI can stabilize the structural integrity of disulfide bonds in Cys48 and Cys77 of growth differentiation factor 15 (GDF15), leading to aberrant function activity of GDF15 and secretion. Interestingly, secreted GDF15 consequently not only further upregulates CSC-related gene expression but also induces the activation of cancer-associated fibroblasts through the transforming growth factor beta receptor type 2 (TGFBR2)–transforming growth factor β (TGFβ)–TGFBI self-regulatory feedback loop to promote stemness in BLCA. TGFBI knockdown or GDF15 inhibition results in a decrease in functional proteins associated with stemness maintenance, which suppresses bladder CSCs’ self-renewal and effectively improves the efficacy of chemotherapy. Together, these findings demonstrate the pivotal role of TGFBI in BLCA’s stemness maintenance and BLCA progression, highlighting that the inhibition of the TGFBI/GDF15 axis is a potential therapeutic strategy for the amelioration of cancer chemotherapy.

Paeoniflorin (PF) is a bioactive monoterpene glycoside with well-established anti-inflammatory properties; however, its therapeutic potential and molecular mechanism in nephrolithiasis remain insufficiently defined. In this study, network pharmacology was employed to predict potential targets of PF. In vitro, HK-2 cells exposed to calcium oxalate (CaOx) crystals were treated with PF to evaluate inflammatory responses, crystal adhesion, and NF-κB activation. A glyoxylic acid-induced mouse model was used to assess renal crystal deposition, renal function, and expression of key regulatory targets, supported by transcriptomic profiling. Molecular docking and molecular dynamics simulations were performed to analyze the interaction between PF and Serpine1, and functional relevance was further examined using exogenous Serpine1 rescue experiments. Integrated analyses indicated that PF exerts anti-nephrolithic effects predominantly through modulation of inflammation-associated signaling. PF dose-dependently suppressed CaOx-induced NF-κB phosphorylation and nuclear translocation, reduced the release of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β), and decreased crystal adhesion in HK-2 cells. In vivo, PF markedly attenuated renal crystal deposition, improved renal functional indices, and downregulated renal Serpine1 and IL-1β expression. Computational simulations supported a stable interaction between PF and Serpine1 (binding energy − 6.22 kcal/mol). Consistently, supplementation with exogenous Serpine1 partially reversed PF-mediated inhibition of NF-κB activation and cytoprotective effects. Collectively, these findings suggest that PF mitigates CaOx-associated renal injury by suppressing inflammation-centered signaling programs involving the Serpine1/NF-κB axis, thereby providing mechanistic insight into PF as a potential multi-target therapeutic candidate for nephrolithiasis.

Background: Aberrant glycosylation is closely associated with tumor progression, changes in the tumor microenvironment, and chemoresistance. This study aimed to identify prognostic sialylation-related genes in bladder cancer and define the role of ST3GAL6 in gemcitabine-cisplatin resistance. Methods: Molecular subtype analysis, prognostic analysis, and risk model construction were performed for sialylation-related genes using transcriptomic data and clinical information from the TCGA database. GC-resistant bladder cancer cell models were established for transcriptomic sequencing and untargeted metabolomic analysis. Cell proliferation and drug sensitivity assays were performed to evaluate the function of ST3GAL6. The regulatory relationship between IGF2BP3, ST3GAL6, and the PI3K pathway was further assessed by combining database analysis with molecular experiments. Results: Sialylation-related molecular patterns were associated with patient prognosis and tumor microenvironment features, particularly fibroblast-related characteristics, in bladder cancer. The key model gene ST3GAL6 was upregulated in bladder cancer tissues and was closely associated with prognosis. In GC-resistant bladder cancer cells, ST3GAL6 expression was significantly increased and accompanied by enhanced sialylation activity. ST3GAL6 promoted bladder cancer cell proliferation and reduced sensitivity to cisplatin and gemcitabine, at least in part through the PI3K-AKT-mTOR pathway. IGF2BP3 was also upregulated in resistant cells, is positively correlated with ST3GAL6, and may help maintain ST3GAL6's expression by stabilizing its mRNA. Conclusions: Our findings suggest that aberrant sialylation is involved in bladder cancer progression and GC resistance. The IGF2BP3-ST3GAL6-PI3K/AKT/mTOR signaling axis may contribute to this process and may serve as a potential biomarker and therapeutic target in bladder cancer.

Background Tertiary lymphoid structures (TLSs) are associated with prognosis and immunotherapy response in clear cell renal cell carcinoma (ccRCC), but their assessment requires invasive tissue sampling. This study aimed to develop and validate a CT-based multi-modal artificial intelligence model for preoperative TLS prediction in ccRCC and to uncover the pathological and molecular determinants of the model’s predictions. Methods This multicenter, retrospective-prospective study enrolled 1,361 patients with ccRCC from eight tertiary hospitals, assigned to primary, test 1, test 2, prospective, multi-omics and immune checkpoint inhibitor (ICI) therapy sets. Arterial-phase CT images were used to train 2D, 2.5D, and 3D deep learning (DL) networks, alongside machine learning (ML) radiomics models. The best single-modality models were integrated via pre-fusion (PFM) and late-fusion (LFM) strategies. Performance was evaluated via the area under the receiver operating characteristic (ROC) curve (AUC). Prognostic value was analyzed via Kaplan-Meier curves. Interpretability was investigated using SHaply Additive Explanations (SHAP) algorithm, HoVer-Net cell segmentation, and bulk RNA sequencing analysis. Results Among the 22 single-modality models, 2.5D-DenseNet161, 2D-ResNet50, Radiomics-XGBoost, and 3D-ResNet101 performed best in test set 1 (AUCs: 0.899, 0.888, 0.878, and 0.756, respectively), which were adopted for fusion models’ establishment. The SVM-based LFM surpassed the PFM (AUC: 0.938 vs. 0.919) and maintained stability across the test 2, prospective, and multi-omics sets (AUC range, 0.877–0.918). LFM-predicted TLS positive significantly correlated with prolonged progression-free survival (all P < 0.05) and effectively discriminated clinical responders from non-responders for ICI therapy (AUC: 0.807). Model predicted TLS positive showed higher inflammatory cell densities (P < 0.05), upregulation of the CXCL13-CXCR5 axis and B-cell-related markers, enrichment of humoral immune response pathways and increased naive and memory B-cell infiltration (P < 0.05). Conclusions The multi-modal LFM provides a non-invasive surrogate for TLS assessment in ccRCC and may support preoperative risk stratification and immunotherapy decision-making.

This study investigated the biological functions and molecular mechanisms of RNA-binding motif protein 11 (RBM11) in bladder cancer (BCa) progression. Integrated bioinformatics analysis of the TCGA database and validation in clinical tissues revealed that RBM11 is significantly upregulated in BCa and positively correlated with advanced tumor stage, poor prognosis, and epithelial-mesenchymal transition (EMT). RBM11 knockdown effectively suppressed migration, invasion, proliferation, and chemoresistance of BCa cells, whereas RBM11 overexpression produced opposite effects. Mechanistically, RBM11 promotes GNPDA1 expression by regulating alternative splicing of GNPDA. Furthermore, GNPDA1 directly interacts with PKM2 and inhibits its ubiquitin-proteasome-mediated degradation, thereby stabilizing PKM2 protein levels, enhancing glycolysis, and promoting malignant progression of BCa. Collectively, these findings indicate that RBM11 drives malignant progression of BCa through the GNPDA1-PKM2 axis, enhancing glucose metabolism reprogramming and EMT process, suggesting that RBM11 may be a potential therapeutic target for BCa.

Introduction: Hormone-sensitive prostate cancer typically progresses to a castrationresistant stage after an average of 18-24 months of androgen deprivation therapy (ADT). The anti-apoptotic factor X-linked inhibitor of apoptosis protein (XIAP) is not only implicated in the development of prostate cancer but also plays a critical role in its progression to the castration-resistant state. Objective: This study aims to investigate the relationship between ADT and the regulation of XIAP. Methods: The protein expression levels of midline 1 (MID1), serine/threonine kinase proviral integration site for Moloney murine leukemia virus 2 (PIM2), protein phosphatase 2A (PP2A), eukaryotic translation initiation factor 4B (EIF4B), phosphorylated EIF4B, and XIAP were analyzed and compared among castrationna & iuml;ve, hormone-sensitive, and castration-resistant prostate cancer (CRPC) tissues. The MID1 gene was manipulated in both androgen-dependent and androgenindependent prostate cancer cells to evaluate its effect on XIAP protein expression and the apoptosis rate of the prostate cancer cells. Results: XIAP protein expression and EIF4B phosphorylation levels were significantly increased in CRPC. In contrast, PIM2 and EIF4B protein expression levels remained similar before and after the development of castration resistance. MID1 protein expression level was significantly elevated, while PP2A expression was significantly reduced in CRPC tissues. Conclusion: ADT may lead to elevated MID1 and reduced PP2A protein expression levels, which indirectly enhance the phosphorylation activity of PIM2 on EIF4B, thereby increasing XIAP expression and reducing apoptosis in prostate cancer cells. This mechanism likely contributes to disease progression toward the castration-resistant stage.

BackgroundBladder urothelial carcinoma (BLCA) exhibits heterogeneous outcomes, creating an urgent need for reliable prognostic biomarkers. Glycosylation modifications are crucial in cancer but understudied for BLCA stratification.MethodsUsing clinical and transcriptomic data from The Cancer Genome Atlas (TCGA) and glycosylation-related genes from the Gene Set Enrichment Analysis (GSEA) database, we constructed a prognostic signature via LASSO regression. It was validated using receiver operating characteristic (ROC) curve and stratified survival analyses. The key gene, alpha-1,3-mannosyltransferase (ALG3), was experimentally validated.ResultsA novel 9-glycosylation-mRNA signature effectively stratified BLCA patients into distinct risk groups with significant overall survival differences. The model showed robust predictive accuracy (AUC) and remained independent of common clinicopathological factors. We identified ALG3 as central to the signature, confirming its elevated tumor expression and critical role in promoting cancer cell proliferation.ConclusionWe established a potent, glycosylation-based prognostic model for BLCA. Functional validation of ALG3 underscores glycosylation's biological importance in tumor progression and highlights its therapeutic potential.

Persistent androgen receptor (AR) signaling, metabolic alterations, and changes in the immune microenvironment are contributing factors to the progression of prostate cancer (PCa); however, therapeutic strategies capable of simultaneously targeting these interconnected vulnerabilities remain limited. Here, we identify a combinatorial regimen consisting of dihydroartemisinin and eupatilin (D&E) as a suppressor of PCa progression through coordinated disruption of tumor-intrinsic survival signaling and protumor immune interactions. Functionally, D&E suppressed the proliferation of PCa cells. Mechanistically, D&E induces ferroptosis in PCa cells, as evidenced by lipid peroxidation, intracellular iron accumulation, malondialdehyde elevation, and glutathione reduction. We further identified AR as an important target of D&E and found that it promotes resistance to ferroptosis by sustaining the SLC7A11 antioxidant axis through transcriptional activation of SLC7A11. Upstream, D&E inhibited the NF-κB subunit p65, which transcriptionally maintains AR expression, thus establishing an NF-κB/AR/SLC7A11 signaling cascade underlying ferroptosis resistance in PCa. Beyond tumor-intrinsic effects, D&E also remodels the tumor microenvironment. Specifically, AR-high tumor cells were found to facilitate M2-like macrophage polarization. In turn, M2-like macrophages could secrete SPP1, which potentially augments malignant properties and AR signaling in tumor cells via the CD44 pathway, thereby constituting a protumorigenic positive feedback loop. Notably, D&E attenuated the M2-like phenotype, reduced SPP1 secretion, and reduced macrophage-mediated promotion of PCa proliferation. In immunocompetent mouse models, D&E further synergized with anti-PD-L1 to suppress PCa progression. Together, these findings indicate that D&E exert antitumor effects in PCa by promoting ferroptosis-associated cell death and modulating macrophage-associated microenvironmental signaling, with potential therapeutic relevance particularly in AR-positive PCa.

Resistance to enzalutamide (Enza) in castration-resistant prostate cancer (CRPC) is linked to poor prognosis. While KDM5B is highly expressed in Enza-resistant CRPC, the mechanisms of resistance remain poorly understood. We applied an integrated approach to study KDM5B using bioinformatics analyses of single-cell and multi-omics data, along with in vitro and in vivo validation. We explored mechanisms through lactylation proteomics, CRISPR/Cas9 editing, ChIP, and dual-luciferase reporter assays. KDM5B induces Enza resistance by epigenetically suppressing PTEN, which in turn activates the PI3K/Akt signaling pathway to upregulate PGK1 and drive metabolic reprogramming and lactate production. Lactate acts as a substrate for p300-mediated lactylation of hnRNPA1 at lysine 179 (K179), stabilizing hnRNPA1 by blocking NEDD4L-mediated ubiquitination and promoting AR-V7 splicing. A potential positive feedback loop enhances this effect: KDM5B activates AR, and AR, in turn, increases KDM5B expression. Inhibiting KDM5B or p300 can reverse Enza resistance in vivo. We identify a mechanism linking metabolism, epigenetics, and a KDM5B/AR feedback loop in drug resistance. These findings suggest that multi-target strategies may represent a promising approach to overcome Enza resistance in CRPC. • KDM5B is a histone demethylase that is upregulated in enzalutamide resistant prostate cancer. • KDM5B drives tumor metabolic reprogramming by activating PI3K/Akt pathway through epigenetic inhibition of PTEN, which in turn transcriptionally up-regulating PGK1. • Lactate causes lactylation of lysine at position 179 of hnRNPA1, leading to abnormal splicing of AR and upregulation of AR-V7. • p300/HDAC1/HDAC2 jointly regulate the lactylation process of hnRNPA1. • The ligand-independent AR signaling pathway positively feedback promotes the upregulation of KDM5B expression.

Background: Lymph node (LN) metastasis is the leading cause of unfavorable prognosis in bladder cancer (BCa), which involves a highly complex tumor microenvironment. The detailed molecular mechanisms that drive BCa LN metastasis, however, are not yet fully elucidated. Methods: In this study, single-cell transcriptomic profiling of 153,339 cells from seventeen BCa patients with or without LN metastases was analyzed. We integrated single-cell transcriptomic data and bulk RNA sequencing data, and employed multiple strategies for the analysis and identification of heterogeneity in epithelial and macrophage cells in BCa. Furthermore, mouse model and cell experiments were performed to validate and dissect the characteristics of the immunosuppressive microenvironment for LN metastases. Results: This study found that DNMT1(+) epithelial cells, characterized by a high potential for epithelial-mesenchymal transition, exhibited a propensity for LN metastasis. Notably, a specific subpopulation of tumor-associated macrophages (TAMs), marked by selective expression of SELENOP, has been identified. Intercellular crosstalk analysis observed the CXCL signal flow directed primarily from DNMT1(+) epithelials to SELENOP+ TAMs. Through both in vitro and in vivo validation, it was suggested that DNMT1 promotes the recruitment and M2-type polarization of TAMs by enhancing CXCL17 expression and secretion. Additionally, CXCL17-mediated reprogramming of TAMs is associated with increased lymphangiogenesis and LN metastasis in BCa. Mechanistically, DNMT1 was found to bind the PTP1B promoter and enhance methylation enrichment at this region, which is consistent with reduced PTP1B expression and activation of JAK2/STAT3 signaling, thereby promoting the expression and secretion of CXCL17. Conclusions: These findings uncover key molecular mechanisms that shape the lymphatic metastatic niche and offer a foundation for novel strategies aimed at targeting pro-metastatic elements to suppress BCa progression and metastasis.

Deprivation of nutrients in the tumor microenvironment drives malignant progression, yet the molecular mechanisms linking metabolic stress to metastasis in bladder cancer remain incompletely understood. Here, we report that nutrient-deprivation stress promotes metastasis by orchestrating a post-translational modification cascade centered on Beclin-1. Clinical analysis revealed that acetylation of Beclin-1 at lysine residues K430 and K437 was significantly reduced in muscle-invasive bladder cancer (MIBC) compared with non-muscle-invasive bladder cancer (NMIBC), a molecular signature inversely correlated with elevated phospho-eIF2α, a marker of cellular starvation. Mechanistically, nutrient deprivation dynamically regulates the expression of the deacetylase SIRT1 and acetyltransferase p300, shifting the balance toward Beclin-1 deacetylation. This deacetylation event serves a dual function: it enhances Beclin-1 protein stability by shielding it from TRIM21-mediated K48-linked ubiquitination and proteasomal degradation, and it promotes autophagosome formation by strengthening its interaction with pro-autophagic partners VPS34, ATG14, and UVRAG while weakening its binding to the inhibitor Rubicon. Consequently, this leads to sustained autophagy activation and epithelial-mesenchymal transition. Genetic and pharmacological interventions further confirmed the central role of this axis, demonstrating that SIRT1 activation by resveratrol promoted metastasis, whereas p300 activation by CTB suppressed it. Crucially, these effects were abrogated in cells expressing deacetylation-mimetic Beclin-1 mutants, suggesting a direct causal link. Our study unveils the SIRT1/p300-Beclin-1-TRIM21 axis as a key nutrient-sensing pathway that promotes bladder cancer metastasis through crosstalk between acetylation and ubiquitination. These findings identify new therapeutic vulnerabilities in advanced bladder cancer.

Neoadjuvant immunochemotherapy (nICT), defined in this study as PD-L1 blockade combined with cisplatin-based chemotherapy, provides substantial clinical benefits in muscle-invasive bladder cancer (MIBC). However, the tumor immune microenvironment (TIME) is highly heterogeneous, resulting in variable patient responses and persistent therapy resistance. This study investigates key immune cell subsets, intercellular communication networks, and spatial distributions within the TIME of MIBC, with a focus on cellular and spatial features associated with nICT response. Patients were classified into responder and non-responder groups according to RECIST 1.1-based radiological evaluation. Responders were defined as patients achieving complete or partial response, whereas non-responders were defined as patients with stable or progressive disease after adequate nICT exposure and evaluable imaging. Representative computed tomography images and fresh tissue samples were collected from four no-treatment controls (NT), three nICT responders (R), and three nICT non-responders (NR). Samples underwent single-cell RNA sequencing and spatial transcriptomics sequencing. Cell populations were annotated to assess infiltration abundance, cell–cell communication networks, and pseudotime trajectories. Resistance-associated cell subsets and their spatial niches were identified. Following quality control, the tumor microenvironment (TME) was classified into ten major cell subsets based on canonical markers. Cross-sectional group comparative analyses showed significant remodeling of the TME in the NR and R groups. B-cell infiltration was elevated in the NR group, whereas macrophages, fibroblasts, and mast cells were enriched in the R group. Functional alterations in activated CD4 + T cells and impaired differentiation of naive CD8 + T cells were identified as key drivers of acquired therapy resistance. Accumulation of macrophage-derived CXCL8 was also observed as a resistance driver. Cancer-associated fibroblasts (CAFs) served as a physical barrier, contributing to the establishment of an immunosuppressive TME. Notably, strong spatial co-localization between macrophage-derived CXCL8 and CAFs was observed in NR tumors, suggesting a cooperative role in mediating resistance to nICT. Further cell–cell communication analysis using spatial transcriptomics indicated that SPP1 signaling from macrophages (CXCL8) to CAFs contributes to TME reprogramming and resistance to nICT. The integrated analysis identifies distinct immunological features that underlie differential nICT responses in MIBC. These findings provide a theoretical basis for optimizing personalized neoadjuvant therapy strategies.

BACKGROUND:Bladder cancer (BCa) represents the most frequently malignancies of the urinary system with high recurrence rates and heterogeneous outcomes. While peptidyl-prolyl cis-trans isomerase NIMA-interacting 1 (PIN1) is recognized as an oncogene in multiple cancers, its expression pattern, biological function, and the molecular basis of bladder tumorigenesis is still poorly understood. METHODS:We performed comprehensive single-cell RNA sequencing (scRNA-seq) analysis to characterize the cellular heterogeneity of BCa tumor microenvironment and identify malignant epithelial cells. Through integrative bioinformatics approaches combining differential expression analysis, survival correlation, and cellular senescence association, PIN1 was identified as a key downregulated gene. We systematically validated PIN1 expression in clinical specimens and cell lines using immunohistochemistry, Western blot, and qRT-PCR. Functional consequences of PIN1 manipulation were assessed through in vitro assays measuring proliferation, apoptosis, migration, invasion, and cellular senescence. A xenograft mouse model was established to evaluate tumor growth and senescence induction in vivo. RNA sequencing and pathway enrichment analysis were conducted to explore the molecular mechanisms. RESULTS:PIN1 was significantly downregulated in malignant epithelial cells of BCa tissues compared to normal counterparts. Low PIN1 expression correlated strongly with poor overall survival (HR = 0.693, p = 0.0265) and progression-free survival (HR = 0.698, p = 0.0471). Elevated PIN1 expression markedly reduced the proliferative, clonogenic, migratory, and invasive capacities of bladder cancer cells, while concurrently driving apoptosis and senescence. Conversely, PIN1 depletion intensified malignant behavior. In vivo studies further showed that PIN1 up-regulation restrained tumor expansion and elevated senescence-associated β-galactosidase activity. Mechanistically, PIN1 overexpression activated the interferon response pathway, upregulating key components including IFNAR1, IRF7, and IRF9. Rescue experiments confirmed that blockade of the interferon pathway partially reversed PIN1-induced growth suppression and senescence, indicating that interferon signaling is a critical downstream mediator of PIN1's tumor-suppressive effects. CONCLUSION:Our study reveals PIN1 as a previously unrecognized bladder-cancer tumor suppressor that halts progression by provoking senescence and re-awakening interferon signaling. This work rewrites the molecular landscape of the disease and positions PIN1 as both a prognostic indicator and a druggable node for future therapies.

Background Hypothermic oxygenated perfusion (HOPE) is a promising technology to improve donated after cardiac death (DCD) liver graft. It was found that protein phosphatase 2A (PP2A) could regulate autophagy and apoptosis, which play a pivotal role in hepatic ischemia reperfusion injury (IRI). In this study, we aim to explore whether PP2A take part in the mechanism that reduces organ damage after HOPE. Method Adult male Sprague Dawley rats were divided into four groups at random. DCD livers of HOPE group were preserved in a HOPE system after 23 hours of cold storage (CS). All groups’ livers were reperfused in an isolated perfused rat liver (IPRL) system for 1 hour at 37°C. After reperfusion, markers related to IRI and protein expression of PP2A related pathway were examined. BRL-3A cells were cultured and incubated with different concentrations H2O2 (0, 50 μM and100 μM). The cellular production of Reactive Oxygen Species (ROS) was detected via the fluorescent intensity of 2,7-Dichlorodihydrofluorescein diacetate (DCFH-DA), and PP2A related pathway protein expression was measured. Results HOPE group suffered the lighter IRI when compared with CS group, evidenced by the lower hepatocytes injury degree, apoptosis rate, and oxidative stress. Further, compared with CS group, the PP2A and ERK1/2 related autography pathway activation of HOPE group was higher, while the JNK and p38 related apoptosis pathway was down-regulated. Cellular experiment showed that mild oxidative stress (50 μM H2O2) could activate the expression of PP2A and autography pathway protein. Severe oxidative stress (100 μM H2O2) shown the opposite regulation effect. Conclusion Through reducing oxidative stress, HOPE attenuates IRI to rat DCD livers via activating PP2A related autography pathway and inhibiting apoptosis pathway.

ObjectivesPrecise perioperative risk stratification for upper tract urothelial carcinoma (UTUC) is essential. We developed a multimodal prognostic model integrating perioperative clinical data, radiomics, and deep learning (DL) features from baseline CT urography to improve survival prediction and guide adjuvant management.Materials and methodsWe retrospectively enrolled 623 patients from six institutions, divided into training, internal validation, and independent external validation sets. Four single-modal models (clinical, radiomics, 2D DL, and 2.5D DL) were developed, and an integrated combined model was constructed by fusing their prognostic scores. Performance was evaluated using the C-index, area under the curve (AUC), calibration curves, and decision curve analysis (DCA).ResultsThe combined model consistently outperformed all single-modal models across all cohorts. C-indices reached 0.758 (95% CI: 0.712-0.804), 0.725 (95% CI: 0.651-0.798), and 0.704 (95% CI: 0.631-0.777) in the training, internal validation, and external validation sets, respectively, numerically surpassing the best single-modal models. Notably, our 2.5D DL model (C-index: 0.705) demonstrated a consistent incremental improvement over the 2D DL model (C-index: 0.681) in capturing prognostic information. In external validation, the combined model achieved a 3-year AUC of 0.766. DCA indicated the comprehensive model exhibited excellent calibration and provided the highest net benefits.ConclusionThis multimodal system, featuring a robust 2.5D DL strategy, improves overall survival prediction in UTUC. It offers a valuable tool for accurate perioperative risk stratification immediately after radical nephroureterectomy, demonstrating particularly reliable value for 3-year intermediate-term clinical decision-making.Critical relevance statementThis multimodal system advances clinical radiology by fusing perioperative clinical data, radiomics, and DL features from CTU images, enhancing risk stratification accuracy to guide postoperative adjuvant management for UTUC.Key PointsCurrent prognostic models for UTUC lack accuracy, creating an unmet clinical need for precise perioperative risk stratification to guide adjuvant management.A multimodal prognostic model fusing clinical, radiomic, and DL features from baseline CT urography consistently outperformed single-modality models in predicting overall survival.

Background Postoperative recurrence prediction in localized renal cell carcinoma (LRCC) remains clinically challenging. Circulating tumor cells (CTCs) have emerged as promising, minimally invasive biomarkers for disease monitoring and prognostication. This study aimed to develop and interpret a machine learning model to predict recurrence risk in LRCC, and to translate the model into a clinically applicable scoring system. Methods A multicenter retrospective cohort of 326 patients with LRCC, collected from 14 hospitals across 7 provinces in China, was randomly divided into training and validation sets (7:3 ratio). Baseline clinicopathological characteristics and CTC subtype counts/changes were collected. Highly correlated features were removed prior to modeling. Key predictors were selected using LASSO, random forest importance, and recursive feature elimination. Six machine learning algorithms—logistic regression, random forest, support vector machine, XGBoost, naive Bayes, and multilayer perceptron—were trained with fivefold cross-validation, and performance was evaluated by AUC. The best-performing model was interpreted using SHapley Additive exPlanations (SHAP) and translated into a simple threshold-based clinical score integrating five variables (ΔMCTC, MCTC, EpiCTC, ΔEpiCTC, and RENAL score). Patients were stratified into low- (0–2) and high-risk (3–5) groups, and Kaplan–Meier curves compared recurrence-free survival. The predictive accuracy of the new score was further compared with UISS and SSIGN using 1- and 5-year ROC analyses in the validation cohort. Results Among all models, the RF model achieved the highest predictive performance (AUC = 0.850). SHAP analysis identified changes in MCTCs and epithelial CTCs as the most critical predictors, followed by baseline MCTCs, epithelial CTCs, and RENAL score. Using RF-derived optimal thresholds, patients received 1 point for each variable exceeding its cutoff and 0 otherwise. Total scores stratified patients into low-risk (0–2 points) and high-risk (3–5 points) groups, with the high-risk group showing significantly shorter PFS compared with the low-risk group (p < 0.001). The clinic risk score outperformed conventional prognostic scores, including UISS and SSIGN, in terms of AUC in the validation cohort. Conclusions A machine learning model integrating CTC metrics and anatomical factors accurately predicted LRCC recurrence and outperformed existing prognostic systems. Its simplified, threshold-based clinical score offers a practical approach for individualized postoperative risk assessment. Trial registration: This study was approved by the Scientific Ethics Committee of the Department of Medicine of Xi’an Jiaotong University (No. 2021033) and the number of China Clinical Research Registration is ChiCTR2000035394.

Precise survival risk stratification for bladder urothelial carcinoma (BUC) remains a clinical challenge. We developed and validated a multimodal AI agent that integrates textual, radiographic, and pathological data from 1185 patients across four medical centers to predict survival risk. The agent employs LLMs to standardize pathology reports, interactive deep learning networks for precise CT image segmentation, and extracts features from CT scans and whole slide images using CTVisionNet and MacroVisionNet. The multimodal fusion framework, MATCH-Net, integrates these features with microscopic pathology information and clinical text embeddings using a multi-head attention mechanism to generate a comprehensive prognostic score. In multi-center validation, MATCH-Net demonstrated robust performance (C-index ranging from 0.836 to 0.874) and effectively stratified patients into high- and low-risk groups, identifying potential candidates responsive to adjuvant chemotherapy. Furthermore, the framework enabled the quantification of novel, interpretable prognostic biomarkers and provides a reliable and clinically applicable solution for personalized BUC prognosis.

OBJECTIVE:This study aimed to investigate the effect and mechanism of arctigenin (ARG) on the sensitization of dacarbazine (DTIC) via the regulation of mitophagy. METHODS:In vitro experiments were conducted to explore the effects of ARG on the biological behavior of melanoma cells, mitochondrial autophagy mediated by PINK1/Parkin, and the role of reactive oxygen species (ROS)-mitochondrial autophagy in the regulation of the biological behavior of melanoma cells by an ROS quenching agent, a mitochondrial autophagy inhibitor, and an activator. The effects of ARG and dacarbazine in nude mice were assessed. RESULTS:CCK8 assays revealed that ARG inhibited the proliferation of the human melanoma cell lines A375 and SK-MEL-2. The observation of submicroscopic structures demonstrated mitochondrial damage. Flow cytometry further verified that ARG induced apoptosis. Western blot analysis revealed that the protein expression levels of cleaved caspase 3 and Bax increased, whereas that of Bcl-2 decreased. In addition, ARG increased ROS levels. LC3II/I, PINK1, and Parkin were increased. ARG-induced apoptosis was related to increased mitochondrial oxidative stress and promoted the occurrence of mitochondrial autophagy. After the addition of the autophagy inhibitor Mdivi-1 or the ROS quencher N-acetylcysteine (NAC), the antiproliferative effect of ARG was markedly attenuated. The expression levels of PINK1, Parkin, LC3II/I, cleaved caspase 3, and Bax were increased, whereas that of Bcl-2 was decreased. The formation of mitochondrial autophagosomes was observed by transmission electron microscopy. ARG inhibited the proliferation and induced the apoptosis of melanoma cells in vivo. CONCLUSION:Autophagy-mediated cell apoptosis was activated through the PINK1/Parkin pathway by ARG, effectively inhibiting the proliferation of human melanoma cells.