BACKGROUND:Molecular subtyping of bladder cancer (BCa) using bulk transcriptomic data is hindered by tumor microenvironment (TME) contamination and intra-tumoral heterogeneity. These factors also contribute to variable responses to immune checkpoint inhibitor (ICI) therapy. Single-cell RNA sequencing offers a high-resolution approach to overcome these limitations by enabling tumor-specific analysis and improving therapeutic target identification. MATERIALS AND METHODS:We constructed a large multicenter single-cell atlas of BCa by integrating nine cohorts comprising 123 samples and 842 127 cells. Tumor epithelial cells ( n = 265 962) were identified via copy number variation (CNV) analysis using inferCNV. Pseudobulk RNA profiles were generated to simulate conventional bulk samples (Pseudobulk all ) and purified tumor profiles (Pseudobulk tumor ), enabling systematic evaluation of bulk-based classifiers. Inter-tumoral and intra-tumoral heterogeneity programs were defined based on transcriptional profiles and validated through in vitro and in vivo experiments. Additionally, we cross-analyzed five immunotherapy datasets encompassing 13 immune response phenotypes to identify molecular features associated with therapeutic resistance and sensitivity. RESULTS:Unsupervised clustering identified 11 major cell types and 55 non-epithelial subtypes, with TME heterogeneity driven by tissue origin and malignancy status. Existing molecular classifiers UNC (University of North Carolina at Chapel Hill) showed compromised concordance between Pseudobulk all and Pseudobulk tumor , particularly in stroma-rich samples. We defined three inter-tumoral (GE inter 2-4) subtypes with distinct CNV and transcription factor signatures that remained stable even at low tumor purity. Seven intra-tumoral (GE intra ) programs captured key biological states, including an EMT/Stem-like module associated with immunosuppression and ICI resistance. Mechanistically, TGF-β1 induced expression of EMT/Stem-like genes (e.g., PTHLH ), while Pthlh knockdown synergized with anti-PD-L1 therapy in murine models by enhancing T cell cytotoxicity without altering T cell infiltration. CONCLUSIONS:This study provides a robust single-cell framework for resolving inter- and intra-tumoral heterogeneity in BCa, overcoming limitations of bulk-based subtyping. The TGF-β1-PTHLH axis contributes to ICI resistance, positioning PTHLH as a promising therapeutic target. These findings offer new insights for improving immunotherapeutic strategies in BCa.

ObjectiveTo explore the efficacy and safety of treatment of intramural ureteral tumors via transurethral resection. MethodsA retrospective analysis was conducted on the clinical data of 26 patients with ureteral cancer who underwent transurethral resection of intramural ureteral tumors at the First Affiliated Hospital of Naval Medical University from May 2020 to May 2023. All patients were treated using the “outer-wedge, inner-rotation” resection technique, with ureteral stents placed intraoperatively. Clinical indicators including operation time, intraoperative blood loss, length of postoperative hospital stay, ureteral stent indwelling duration, intraoperative and postoperative complications, postoperative pathological results, and bladder tumor recurrence rate were analyzed. ResultsAll surgeries were successfully completed. The operation time was 45(35, 60) min, intraoperative blood loss was 5(3, 10) mL, length of postoperative hospital stay was 4(3, 5) d, and the ureteral stent indwelling duration was (33.8±9.7) d. All patients did not experience intraoperative complications, the postoperative hydronephrosis rate was 3.8%. Postoperative pathology confirmed urothelial carcinoma in all cases, including 6 cases of stage pTa, 16 cases of pT1, and 4 cases of pT2a. The follow-up period ranged from 12 to 48 months. Intravesical recurrence occurred in 2 patients, and recurrence rate was 7.7%. There were no tumor-related deaths during the follow-up period. ConclusionsTransurethral resection of intramural ureteral tumors is minimally invasive, resulting in less blood loss, and has low rates of postoperative complications such as hydronephrosis and hydroureter. It achieves favorable surgical outcomes while better preserving renal function.

Bladder cancer (BCa) presents a substantial global health burden, with high rates of recurrence and metastasis that limit the effectiveness of current therapies. New therapeutic strategies are urgently needed. This study introduces a novel nanotherapeutic approach utilizing polydopamine (PDA) nanoparticles to co-deliver cisplatin and miR-205-5p for BCa treatment. Combination therapy reduces the dose-dependent toxicity of cisplatin while enhancing tumor cell cytotoxicity. miR-205-5p targets centromere protein F (CENPF), a key regulator of cancer progression. Overexpression of CENPF in BCa correlates with poor prognosis, and miR-205-5p-mediated suppression of CENPF expression inhibits tumor growth. The PDA-based system combines the DNA-damaging effects of cisplatin with the gene-silencing properties of miR-205-5p, resulting in synergistic antitumor activity. This multimodal strategy enhances therapeutic precision and efficacy, providing a promising solution for BCa treatment with significant clinical potential.

Bladder cancer has been considered as one of the most common urinary malignancies. Growing evidence has indicated that Centromere Protein F (CENPF) is a promising molecular biomarker for many human malignant tumors. However, the role of CENPF in bladder cancer (BC) proliferation, migration, invasion, cell cycle and apoptosis thereof remain unclear. In the present study, high-throughput sequencing technology and bioinformatics analysis were conducted to identify mRNAs profiles in 10 pairs of bladder cancer tissues and adjacent noncancerous tissues. CENPF was overexpression in bladder cancer tissues, and higher in MIBC than NMIBC. We investigated the expression of CENPF in 30 other pairs bladder cancers tissues, and the results were in accordance with the sequencing results. Furthermore, Immunohistochemical staining, showed that strong intensity of CENPF in BC tissues than normal tissues. Increased staining of CENPF was detected of tumor cells in MIBC compared with NMIBC. This suggests that CENPF might be highly expressed in aggressive and invasive tumor cells. Subsequently, in vitro functional experiments also demonstrated that the siRNA interference of CENPF expression significantly weakened the proliferation, migration, invasion and apoptosis of BC cells, and the cells were arrested in the G2/S phase in Cell cycle. Moreover, functional enrichment analyses, lncRNA-miRNA-mRNA, and protein-protein interaction networks revealed that CENPF was potentially involved in carcinogenesis and evolution of bladder cancer. Taken together, these results demonstrated that CENPF may serve as a potential biomarker of tumor occurrence, progression, and even prognosis for bladder cancer. However, further research is needed to be further clarified the pathway mechanisms of CENPF in bladder cancer.

Background:Ureteral tumors are rare and present unique diagnostic and therapeutic challenges. Objective:This study aimed to evaluate the efficacy and safety of transurethral resection (TUR) in treating ureteral tumors located in the intramural segment of the bladder wall. Methods:This retrospective study analyzed the clinical data of 24 patients who underwent TUR for intramural ureteral tumors at Changhai Hospital, Shanghai, China, from May 2020 to September 2023. All patients were treated using an "outer wedge, inner rotation" technique, with ureteral stent placed during the procedure. Results:All surgeries were successfully completed, with a median operative time of 45 min (range: 25-75 min), median intraoperative blood loss of 5 mL (range: 1-30 mL), median post-operative hospitalization of 4 days (range: 3-10 days), mean ureteral stent placement duration of 34.1 ± 10.1 days, and a mean tumor resection diameter of 1.9 ± 0.7 cm. There were no intraoperative complications, while post-operative hydronephrosis occurred in 12.5% of patients. Pathological examination revealed that all tumors were urothelial carcinoma, with five cases of Ta stage, one case of pT1 low-grade, 14 cases of pT1 high-grade, and four cases of pT2a high-grade. The median follow-up period lasted for 28 months (range: 11-38 months), with a tumor recurrence rate of 20.8%. Conclusion:TUR of ureteral tumors in the intramural segment is a minimally invasive procedure associated with low blood loss, reduced post-operative complications, and efficacious tumor control while preserving renal function. However, the possibility of local tumor recurrence remains, necessitating close post-operative surveillance.

Background:Adrenocortical carcinoma (ACC) is an extremely rare and highly invasive malignant tumor. However, there is currently no reliable method to predict the prognosis of ACC. Our objective is to construct a nomogram and a risk classification system to predict the 1-year, 3-year, and 5-year overall survival (OS) of ACC.Methods:We retrieved clinicopathological data of patients diagnosed with ACC in The Surveillance, Epidemiology, and End Results (SEER) database and divided them into training and validation cohorts with a 7:3 ratio. Simultaneously, we collected an external validation cohort from The First Affiliated Hospital of Naval Medical University (Shanghai, China). Univariate and multivariate Cox analyses were performed to identify relevant risk factors, which were then combined to develop a correlation nomogram. The predictive performance of the nomogram was evaluated using the concordance index (C-index), receiver-operating characteristic curve (ROC), and calibration curves. Decision curve analysis (DCA) was applied to assess the clinical utility of the nomogram. In addition, Kaplan-Meier survival curves were generated to demonstrate the variation in OS between groups.Results:The final nomogram consisted of five factors: age, T, N, M, and history of chemotherapy. Our prognostic model demonstrated significant discriminative ability, with C-index and the area under the receiver operating characteristic (AUC) values exceeding 0.70. Additionally, DCA validated the clinical utility of the nomogram. In the entire cohort, the median OS for patients in the low- and high-risk groups was 70 and 10 months, respectively.Conclusions:A nomogram and a corresponding risk classification system were developed in order to predict the OS of patients diagnosed with ACC. These tools have the potential to provide valuable support for patient counseling and assist in the decision-making process related to treatment options.

AbstractObjectiveWe have developed explainable machine learning models to predict the overall survival (OS) of retroperitoneal liposarcoma (RLPS) patients. This approach aims to enhance the explainability and transparency of our modeling results.MethodsWe collected clinicopathological information of RLPS patients from The Surveillance, Epidemiology, and End Results (SEER) database and allocated them into training and validation sets with a 7:3 ratio. Simultaneously, we obtained an external validation cohort from The First Affiliated Hospital of Naval Medical University (Shanghai, China). We performed LASSO regression and multivariate Cox proportional hazards analysis to identify relevant risk factors, which were then combined to develop six machine learning (ML) models: Cox proportional hazards model (Coxph), random survival forest (RSF), ranger, gradient boosting with component‐wise linear models (GBM), decision trees, and boosting trees. The predictive performance of these ML models was evaluated using the concordance index (C‐index), the integrated cumulative/dynamic area under the curve (AUC), and the integrated Brier score, as well as the Cox–Snell residual plot. We also used time‐dependent variable importance, analysis of partial dependence survival plots, and the generation of aggregated survival SHapley Additive exPlanations (SurvSHAP) plots to provide a global explanation of the optimal model. Additionally, SurvSHAP (t) and survival local interpretable model‐agnostic explanations (SurvLIME) plots were used to provide a local explanation of the optimal model.ResultsThe final ML models are consisted of six factors: patient's age, gender, marital status, surgical history, as well as tumor's histopathological classification, histological grade, and SEER stage. Our prognostic model exhibits significant discriminative ability, particularly with the ranger model performing optimally. In the training set, validation set, and external validation set, the AUC for 1, 3, and 5 year OS are all above 0.83, and the integrated Brier scores are consistently below 0.15. The explainability analysis of the ranger model also indicates that histological grade, histopathological classification, and age are the most influential factors in predicting OS.ConclusionsThe ranger ML prognostic model exhibits optimal performance and can be utilized to predict the OS of RLPS patients, offering valuable and crucial references for clinical physicians to make informed decisions in advance.

Purpose: Evidence of an association between leukocyte telomere length (LTL) and prostate cancer (PCa) is accumulating; however, their shared genetic basis remains unclear. Materials and Methods: Using summary statistics obtained from the genome-wide association study (GWAS), we quantified the global and local genetic correlations between two traits. Subsequently, we identified potential pleiotropic loci, common tissue-enriched regions, and risk gene loci while inferring assumed causal relationships. Results: Our study demonstrated a global genetic correlation between LTL and PCa (genetic correlation=0.066, p=0.017), which was further confirmed in local genomic regions. Cross-trait GWAS meta-analysis revealed 44 shared loci, including 10 novel pleiotropic single nucleotide polymorphisms appearing concurrently in significant local genetic correlation regions. Notably, two new loci (rs9419958; rs3730668) were additionally validated to co-localize. For the first time, we identified a significant shared genetic enrichment of both traits in the small intestine tissue at the terminal ileum, with functional genes in this region affecting both LTL and PCa. Concurrently, Mendelian randomization analysis indicated a positive causal relationship between LTL and PCa. Conclusions: In conclusion, our study makes a significant contribution to the ongoing debate concerning the potential association between longer LTL and a higher risk of PCa. Additionally, we provide new evidence for the development of therapeutic targets for PCa and propose new directions for future risk prediction in this regard.

Comparison between the results of UroCAD and cytology tests in training cohort.

Comparison between the results of UroCAD and urine cytology tests in the training group.

Abstract Objective:The epidemiological evidence regarding the causal relationship between hyperthyroidism, subacute thyroiditis (SAT), and tumors remains inconclusive. Therefore, we conducted a two-sample bidirectional Mendelian randomization (MR) study to establish the causal relationship between these conditions. Methods: We conducted a bi-directional MR study using publicly available GWAS summary statistics to explore the causality between genetically predicted hyperthyroidism, SAT and the risk of 20 common cancers. The analysis was performed using inverse-variance weighted (IVW), MR-Egger, weighted median, simple mode, and weighted mode methods. The primary results were based on the IVW (random effects), followed by sensitivity analysis. Furthermore, maximum likelihood, penalized weighted median and IVW (fixed effects) were used to confirm the robustness of the findings. Results:IVW analysis revealed a significant positive causal association between hyperthyroidism and breast cancer (OR = 2.20E+05, 95% CI: 7.7733–6.23E+09, P = 0.0187), ovarian cancer (OR =1.0949, 95% CI: 1.0250–1.1696, P = 0.0071), thyroid cancer (OR =3.05E+11, 95% CI: 1.06E+01–8.84E+21, P = 0.0314), and colorectal cancer (OR =1.1345, 95% CI: 1.0293–1.2505, P = 0.0110) ; while hyperthyroidism had an inverse association with bladder cancer (OR =0.9446, 95% CI: 0.9017–0.9896, P = 0.0164), prostate cancer (OR =0.6174, 95% CI: 0.4879–0.7813, P = 5.97E-05), liver and bile duct cancer (OR =0.9723, 95% CI: 0.9540–0.9910, P = 0.0038), brain cancer (OR =0.9699, 95% CI: 0.9460–0.9945, P = 0.0166), and malignant neoplasm of male genital organs (OR =0.8593, 95% CI: 0.7868–0.9385, P = 0.0007). Furthermore, the IVW analysis supported a positive causal relationship between SAT and endometrial cancer (OR =1.031, 95% CI: 1.0032–1.0596, P = 0.0288), while SAT had an inverse association with kidney cancer (OR =0.9015, 95% CI: 0.8255–0.9844, P = 0.0209) and thyroid cancer (OR =0.9143, 95% CI: 0.8390–0.9962, P =0.0407). However, we only observed an inverse association between malignant neoplasm of male genital organs and SAT in the reverse MR analyses. Conclusions: The current investigation offers genetic evidence that hyperthyroidism could potentially elevate the risk of developing breast, ovarian, thyroid, and colorectal cancers. Likewise, SAT is identified as a possible risk factor for endometrial cancer. In light of these findings, further studies are necessary to uncover the underlying mechanisms that establish causal links between hyperthyroidism, SAT, and malignancies.

Video S1. Amphisbaena procedure. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Summary of related data of UroCAD assay.

Supplementary Figures and Figure Legends

Comparison of UroCAD and urine cytology tests in the validation cohort.

Comparison of UroCAD results of urine samples stored for 2 hours and 72 hours.

Abstract Objective Previous observational studies have explored the correlation between testosterone and cancer risk. However, the causal association between testosterone and various cancer types in women remains inconclusive. The objective of this Mendelian randomization study is to evaluate the causal links between total testosterone (TT) and bioavailable testosterone (BT) with cancer risk in females. Methods Initially, a rigorous quality control process was employed to identify suitable instrumental single nucleotide polymorphisms (SNPs) associated with the exposure under investigation that exhibited a significant association. The genetic causal relationship between female testosterone levels and the risk of developing cancers was examined through a two-sample Mendelian randomization. Various analytical methods, including inverse-variance weighted (IVW), MR-Egger, weighted median, simple mode, and weighted mode, were applied in the investigation. Key findings were primarily based on the results obtained via IVW (random effects), and sensitivity analyses were conducted to assess the reliability of the obtained results. Furthermore, maximum likelihood, penalized weighted median, and IVW (fixed effects) methods were utilized to further validate the robustness of the results. Results Based on the results of IVW analysis, our study indicated a positive causal relationship between BT and breast cancer (OR = 1.1407, 95%CI: 1.0627–1.2244, P = 0.0015) and endometrial cancer (OR = 1.4610, 95%CI: 1.2695–1.6813, P = 1.22E-06). Moreover, our findings also showed a positive causal association between TT and breast cancer (OR = 1.1764, 95%CI: 1.0846–1.2761, P = 0.0005), cervical cancer(OR = 1.0020, 95%CI: 1.0007–1.0032, P = 0.0077), and endometrial cancer(OR = 1.4124, 95%CI: 1.2083–1.6511, P = 0.0001). Additionally, our results demonstrated a negative causal relationship between BT and ovarian cancer (OR = 0.8649, 95%CI: 0.7750–0.9653, P = 0.0320). However, no causal relationship was found between BT, TT and other types of cancer (corrected P > 0.05). Conclusions This study elucidates the role of testosterone on the development of breast cancer, endometrial cancer, ovarian cancer, and cervical cancer. It also hints at a potential but fragile link between testosterone and bladder cancer, as well as thyroid cancer. Nonetheless, it's worth noting that no statistically significant relationship between testosterone and various other types of cancer in females was identified.

目的:探究非肌层浸润性膀胱癌(NMIBC)患者卡介苗(BCG)膀胱灌注后不良反应与疗效的关系.方法:回顾性分析2014年4月—2021年4月长海医院收治的141例接受BCG灌注的中、高危NMIBC患者的临床资料.中位随访时间22(1～64)个月,平均年龄(66.2±10.0)岁,中位年龄67(40～98)岁.所有患者均行经尿道膀胱肿瘤切除术(TURBT),术后进行规律BCG膀胱灌注.将单个不良反应进行单因素Cox回归分析,探究单个不良反应与疗效的关系.根据不良反应发生的分级、部位及数量的不同分别进行log-rank分析,探究不良反应不同亚组对疗效的预测作用.结果:121例(85.8％)患者出现BCG灌注相关不良反应.根据CTCAE5.0标准分为1级不良反应65例和2级及以上不良反应76例,两组累计复发率分别为9.2％(6/65)和14.5％(11/76),差异无统计学意义(P = 0.485).进一步将不良反应分为无或局部不良反应62例和全身不良反应79例,两组累计复发率分别为9.7％(6/62)和13.9％(11/79),差异无统计学意义(P = 0.575).根据不良反应数量分为1种及以下不良反应47例、2种不良反应35例、3种及以上不良反应59例,三组累计复发率分别为14.9％(7/47)、11.4％(4/35)和10.2％(6/59),差异无统计学意义(P = 0.705).结论:BCG灌注引起不良反应的分级,部位及数量均与灌注疗效无显著关联.仍需前瞻性、大样本量研究以进一步明确BCG灌注后不良反应与其疗效的关系.

BACKGROUND:This retrospective study aimed to evaluate the diagnostic value of early secretory antigenic target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10) in immunoglobulin A nephropathy (IgAN) associated with renal tuberculosis (RT). METHODS:Forty patients with IgAN (IgAN group), 32 patients with RT (RT group), and 52 patients with IgAN associated with RT (IgAN + RT group) were retrospectively selected for this study. A tuberculin skin test (TST) was conducted, and Mycobacterium tuberculosis (MTB) antibody levels were measured. Immunohistochemistry and western blotting were used to determine the expression of ESAT-6 and CFP-10 proteins in renal tissues. RESULTS:The positive results of TST and levels of serum and urinary MTB antibodies were higher in the RT group than in the IgAN + RT group. The expression levels of ESAT-6 and CFP-10 proteins were the highest in the IgAN + RT group and lowest in the IgAN group. The receiver operating characteristic curves indicated that the area under curve (AUC) value of the ESAT-6 protein for the diagnosis of IgAN associated with RT was 0.907 and the cut-off value of the integral optical density (IOD) was 26.72. Diagnosis based on ESAT-6 protein levels showed 75% sensitivity and 94.2% specificity. The AUC value of the CFP-10 protein for the diagnosis of IgAN associated with RT was 0.8 and the cut-off value of IOD was 25.67. Detection based on CFP-10 protein levels showed 63.9% sensitivity and 84.6% specificity. CONCLUSIONS:Our study provides evidence for the potential of ESAT-6 and CFP-10 proteins as candidate markers in the diagnosis of IgAN associated with RT.

Long non-coding RNAs (lncRNAs) can modulate various biological processes and behaviors in most human cancers. LncRNA EIF3J-AS1 has been reported as an oncogene in various tumors, but whether it exerts functions in malignant progression and gene expression in prostate cancer (PCa) remains unknown. In this study, we investigated the high level of EIF3J-AS1 in PCa tissues and cells, and used functional assays to show that knocking down EIF3J-AS1 inhibited PCa cell proliferation and metastatic ability. A preliminary mechanistic investigation also showed that EIF3J-AS1 may increase the expression of MAF bZIP transcription Factor G (MAFG) in PCa. The expression correlation between EIF3J-AS1 and MAFG was found to be positive in PCa tissues. Finally, rescue assays showed that MAFG might be involved in the EIF3J-AS1-mediated malignant phenotype in PCa cells. This study demonstrated that EIF3J-AS1/MAFG may play a key role in facilitating PCa progression.