AbstractBackgroundRecurrent bladder cancer is the most common type of urinary tract malignancy; nevertheless, the mechanistic basis for its recurrence is uncertain. Innovative technologies such as single‐cell transcriptomics and spatial transcriptomics (ST) offer new avenues for studying recurrent tumour progression at the single‐cell level while preserving spatial data.MethodThis study integrated single‐cell RNA (scRNA) sequencing and ST profiling to examine the tumour microenvironment (TME) of six bladder cancer tissues (three from primary tumours and three from recurrent tumours).FindingsscRNA data‐based ST deconvolution analysis revealed a much higher tumour heterogeneity along with TME in recurrent tumours than in primary tumours. High‐resolution ST analysis further identified that while the overall natural killer/T cell and malignant cell count or the ratio of total cells was similar or even lower in the recurrent tumours, a higher interaction between epithelial and immune cells was detected. Moreover, the analysis of spatial communication reveals a marked increase in activity between cancer‐associated fibroblasts (CAFs) and malignant cells, as well as other immune cells in recurrent tumours.InterpretationWe observed an enhanced interplay between CAFs and malignant cells in bladder recurrent tumours. These findings were first observed at the spatial level.

Background The discovery of cuproptosis provides a new way to make full use of the pathophysiological effects of copper for anticancer therapy and could help identify a therapeutic target in bladder cancer. Methods In this study, 411 BLCA tumor samples from the Cancer Genome Atlas (TCGA) cohort were obtained. Differentially expressed genes (DEGs) between chemotherapy-sensitive and chemotherapy-resistant mouse bladder cancers were also obtained. Sixteen genes were defined as cuproptosis-related genes (CRGs), and the cutoff score was calculated based on LASSO Cox regression. CCK-8 and Transwell assays were used to detect the migration and proliferation of 5637 and T24 cells, respectively. Liarozole dihydrochloride (L-D), a mild P450, inhibits the expression of CYP26B1. Multiple immunohistochemistry analyses were used to explore the association between the immune microenvironment and CRGs. Results A higher Cuproptosis score was significantly associated with worse overall survival in BLCA ( p <0.0001, HR=2.32). Single-cell transcriptional data were used to assess the function of CYP26B1 and CYP26B1 may be negatively associated with DNA damage and repair. In vitro experiments indicated that overexpression of CYP26B1 enhanced the migration and proliferation of BLCA cells, while L-D inhibited the migration and proliferation of BLCA cells. A higher expression level of Dihydrolipoamide S-Succinyltransferase ( DLST ) serves could as a risk factor in patients treated with atezolizumab and higher expression of DLST suggest an immunosuppressive microenvironment. Conclusions CYP26B1 may be a therapeutic target for bladder cancer, and the higher expression of DLST may suggest an immunosuppressive microenvironment in BLCA. Key Messages Cuproptosis may be associated with the chemotherapy-resistance of bladder cancer and our findings may provide new ideas for the treatment of bladder cancer.

BACKGROUND:Melatonin is a hormone naturally produced by the pineal gland in the brain. In addition to modulating circadian rhythms, it has pleiotropic biological effects including antioxidant, immunomodulatory, and anti-cancer effects. Herein, we report that melatonin has the ability to decrease the growth and metastasis of androgen-dependent prostate cancer.METHODS:To evaluate the anti-cancer effect of melatonin on androgen-sensitive prostate cancer in vitro or in vivo, the effects of cell proliferation, apoptosis, migration and invasion were analyzed by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), colony formation, flow cytometry, Transwell assay, and immunohistochemistry (IHC), respectively. Next, the interaction between androgen receptor (AR) and SUMO specific protease 1 (SENP1) was detected by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blotting, and confirmed by luciferase reporter assay. Furthermore, the Small Ubiquitin-like Modifier (SUMO) proteins are a group of small proteins that are covalently attached to and detached from other proteins in cells to modify their function. (SUMOylation) of histone deacetylases 1 (HDAC1) was measured by proximity ligation assay (PLA).RESULTS:The treatment of melatonin cripples the transcriptional activity of AR, which is essential for the growth of the androgen-dependent prostate cancer cell, LNCaP. The lower activity of AR was dependent on melatonin induced SUMOylation of HDAC1, which has been established as a key factor for the transcriptional activity of AR. Mechanistically, the effect of melatonin on AR was due to the decreased SENP1 protein level and the subsequent increased HDAC1 SUMOylation level. The overexpression of SENP1 abrogated the anti-cancer ability of melatonin on LNCaP cells.CONCLUSIONS:These findings indicate that melatonin is a suppressor of androgen-dependent prostate cancer tumorigenesis.

Abstract Objective To introduce a new sequential sheath urethral dilatation (SSUD) method to treat male short segment posterior urethral stricture (MSS-PUS). Methods 67 male patients who had a MSS-PUS were enrolled and randomly assigned to one of two groups: group A (SSUD) and group B (Cold knife internal urethrotomy, CIU). Baseline information, including age, diabetes status, location of urethral stricture, preoperative ultrasound post-void residual (PVR) measurement, preoperative maximum urinary flow rate (Qmax), and preoperative International Prostate Symptom Score (IPSS) were collected and compared between groups. The operation time, postoperative length of hospital stay, indwelling catheter size, success rate, operative complications, recurrence, postoperative PVR, Qmax, and IPSS were evaluated and compared between the two groups. Results There was no significant difference in the baseline data, postoperative data, the complication, recurrence rates, postoperative length of hospital stay and postoperative PVR measurements between the two groups (p > 0.05), but the difference in immediate success rate, operative time, indwelling catheter size, postoperative Qmax and postoperative IPSS was statistically significant (p < 0.05). Conclusion We introduced a new SSUD method to treat MSS-PUS. The immediate success rate, operation time, and surgical effect were better than the CID procedure.

The prolyl 3-hydroxylase family member 4 gene (P3H4) is involved in the development of human cancers. The association of P3H4 with bladder cancer (BC) prognosis is unclear. This study aimed to analyze the association of P3H4 with BC prognosis. RNA-Seq data were downloaded from The Cancer Genome Atlas project and BC microarray datasets (GSE13507, GSE31684, and GSE32548) were downloaded from the Gene Expression Omnibus database. We analyzed the differences in P3H4 expression levels between BC tumors and non-tumor tissues and between samples with different clinical information. The association of P3H4 and P3H4-related genes with BC prognosis and the possibility of using P3H4 expression as a prognostic biomarker in BC patients were also analyzed. RevMan was used to perform the meta-analysis. P3H4 was upregulated in BC tissues compared with the adjacent non-tumor tissues (p = 4.06e−08). Univariate Cox regression analysis and meta-analysis showed that high P3H4 expression level contributed to a poor BC prognosis (Hazard ratio, HR = 1.348, 95% CI 1.140–1.594, p = 4.89e−04; meta-analysis: HR = 1.45, 95% CI 1.10–1.91; p = 9.00e−03). Among the genes related to P3H4, the PLOD1 gene was closely associated with P3H4 expression (r = 0.620, p = 2.49e−44). Also, a meta-analysis showed that PLOD1 expression was associated with a poor prognosis in BC patients (HR = 1.77, 95% CI 1.31–2.38; p = 2.00e−04). The P3H4 and PLOD1 genes might be used as reliable prognostic biomarkers for BC.

Background: Thulium laser (Tm:YAG) prostate surgery is a safe and effective procedure with low morbidity and comparable clinical outcomes to those of transurethral resection of the prostate (TURP). However, the sexual function outcomes (erectile and ejaculatory function) have been scarcely studied. Aim: We aimed to assess the impact of Tm:YAG prostate surgery on sexual outcomes (erectile and ejaculatory function) and compare them with those patients undergoing TURP. Material and Methods: We searched digital databases like PUBMED, SCOPUS, CENTRAL and EMBASE using relevant keywords to identify comparative studies on TURP and non-comparative studies on Tm:YAG prostate surgery that assessed sexual outcomes. We performed qualitative and quantitative analyses with the extracted data. We carried out a meta-analysis to compare postoperative International Index of Erectile Function (IIEF-5) scores and incidences of retrograde ejaculation (RE) in patients undergoing either Tm:YAG or TURP. The pre-operative and post-operative IIEF-5 scores were pooled to estimate overall scores. Results: We included 5 comparative and 8 non-comparative studies in this review. We found the postoperative IIEF-5 score improvements to be significantly higher in the Tm:YAG prostate surgery group than in the TURP group with a significant mean difference (MD) of 0.45 (95% CI, 0.18 to 0.72; P= .001). We found no significant associations between the procedures. The pooled OR for the association of RE was estimated at 0.90 (95% CI, 0.50 to 1.60; P= .71; I-2 = 0%). Conclusion: Tm:YAG prostate surgery improves erectile function more than TURP, according to our findings. Tm:YAG prostate aided surgery also outperforms TURP in terms of preserving sexual function following surgery.However, We found similar or no difference in incidence of RE between Tm:YAG prostate surgery and TURP. Copyright (C) 2021 The Authors. Published by Elsevier Inc. on behalf of the International Society for Sexual Medicine.

BACKGROUND:We conducted a two-center study to investigate the prognostic value of preoperative fibrinogen-albumin ratio (FAR) in patients undergoing radical cystectomy (RC).METHODS:The clinical and survival data of 267 patients with bladder cancer (BCa) treated with RC were collected, of which 140 patients from Xuzhou Central Hospital were divided into training set and 127 patients from The Second Affiliated Hospital of Nantong University were divided into validation set. X-tile software was used to obtain the optimal cut-off values for preoperative platelet-to-lymphocyte ratio (PLR), neutrophil-to-lymphocyte ratio (NLR) and FAR. Receiver operating characteristic (ROC) curves were used to compare the predictive ability of PLR, NLR, FAR and modified Glasgow prognostic score (mGPS). Kaplan-Meier curves were used to assess overall survival (OS) and progression-free survival (PFS) of patients in different FAR groups. Univariate and multivariate Cox regression were used to assess patients' independent risk factors, and R software was used to construct prognostic nomograms.RESULTS:In the training set, the optimal cut-off values for PLR, NLR and FAR were 76.76, 3.97 and 0.08, respectively. Both in the training and validation sets, FAR had better ability to predict OS and PFS than PLR and NLR, and patients in the higher FAR group had worse OS and PFS. In the multivariate Cox regression analysis, FAR was an independent risk factor for OS [hazard ratio (HR) 3.569, 95% confidence interval (CI): 1.015-12.546, P=0.047] and PFS [HR 5.071, 95% CI: 1.394-18.451, P=0.014]. In addition, FAR-based prognostic nomograms had high predictive ability than TNM staging.CONCLUSION:Preoperative FAR is an independent prognostic factor for OS and PFS in BCa patients treated with RC, and a high FAR predicted a poor prognosis. In addition, a prognostic nomogram based on FAR can better predict individual survival.

Bladder cancer (BCa) is a common malignancy in the urinary system. Ras-related protein Rab-27A (RAB27A) and Ras-related protein Rab-27B (RAB27B) have been verified to be closely related to the development of many tumors. Since the role of both RAB27A and RAB27B in BCa have not been reported, we intended to explore the function and mechanism of RAB27A and RAB27B in BCa development. Reverse transcription quantitative polymerase chain reaction revealed that RAB27A/B showed high expression in BCa tissues and cells. Cell counting kit-8, wound healing and Transwell assays as well as western blot analyses revealed that silencing RAB27A/B suppressed BCa cell proliferation, migration and invasion as well as the epithelial-mesenchymal transition (EMT) process. Based on bioinformatics analysis and our experiments, microRNA-186-5p (miR-186-5p) was found to be the upstream miRNA of RAB27A/B in BCa. MiR-186-5p expression was significantly downregulated in BCa cells and tissues. MiR-186-5p directly targeted the 3'-untranslated region (3'-UTR) of RAB27A/B and downregulated both mRNA and protein levels of RAB27A/B in BCa cells. MiR-186-5p overexpression suppressed BCa cell proliferation, migration and invasion as well as the EMT process in vitro and inhibited tumor growth in vivo. Overexpressing RAB27A/B rescued the inhibitory effect of miR-186-5p on malignant phenotypes of BCa cells. Furthermore, miR-186-5p inactivated the phosphatidylinositol 3-Kinase (PI3K)/mitogen-activated protein kinase (MAPK) signaling pathway by downregulating the expression of RAB27A/B, as shown in western blot analysis. Overall, miR-186-5p suppressed BCa cell proliferation, migration, invasion and EMT process and inhibited xenograft tumor growth by targeting RAB27A/B to inactivate the PI3K/MAPK signaling.

For the treatment of ejaculatory duct obstruction, transurethral seminal vesiculoscopy (TSV) is the most common method, but the success rate is much lower than studies that have reported. So we developed a new ultrasound-guided seminal vesicle radiography (UGSVR) combining CT three-dimensional reconstruction (CT-TR) technique to improve the success rate of TSV. Between June 2018 and November 2019, 32 patients were enrolled and randomly assigned to two groups: experimental group (UGSvR combining CT-TR) and control group (standard evaluation). Baseline information, including age, smoking history and body mass index (BMI), was compared preoperatively. Surgical parameters included success rates (SR), surgical time (ST), catheter days (CD), length of hospital stays (HS) and complications were compared between groups. There were no statistically significant differences in baseline data between the two groups (all p > .05). There were no significant differences in the CD, HS and complications between the two groups (all p > .05), but the differences in ST and SR were statistically significant (p < .05). In conclusion, this new technique of UGSvR combining CT-TR was achieving a satisfactory increase in the success rate of TSV, while not increasing the incidence of complications, compared to normal evaluation before TSV operation.

Introduction: There have been problems with low qualification operator-related complications and failures of transurethral seminal vesiculoscopy (TSV) in China. Aim: To study the guiding role of seminal tract anatomical study (STAS) in TSV. Material and methods: We performed STAS to study the structure, morphology, duct trajectory, and anatomical relationships between the seminal vesicles and the adjacent tissue in pelvic specimens from 12 adult cadavers. Then the surgical effects and complications of 82 cases of TSV performed by 3 doctors were retrospectively studied to compare the difference between the two groups of before and after the anatomical study. Results: The anatomical studies of the 12 adult cadaveric pelvis specimens identified the lengths and widths of the right- and left-side seminal vesicles and tracts. The TSV can treat lesions located in the distal seminal tract and vesicle, but proximal lesions cannot be reached, which is an anatomical limitation of this technique. There were significant differences in the surgical times and the surgical validity rates between the 2 groups. Conclusions: Our anatomical study of the seminal tract and seminal vesicles is valuable for guiding TSV in clinical practice.

Objective: To introduce a new position for ureteroscopic holmium laser lithotripsy for patients with upper ureteral calculi. Materials and methods: Between June 2014 and May 2017, 192 patients were enrolled in this study. Patients were randomly assigned to one of two groups: group A, ureteroscopic lithotripsy (URSL) in the Trendelenburg position; or group B, URSL in the standard position. Baseline information, including gender, age, body mass index (BMI), stone side, stone size and hydronephrosis grade, was collected and determined preoperatively. Stone-free rate (SFR) was evaluated 3 weeks after surgery and was defined by the absence of residual stones or the presence of residual stones <2 mm in diameter. Operation time, hospital stay, stone migration, operative complications and SFR were assessed and compared between the two groups. Results: There were no statistically significant differences in gender, age, BMI, stone side, stone size, serum creatinine or hydronephrosis grade between the two groups (all p > 0.05). There were no significant differences in the postoperative hospital stay or postoperative complications between the two groups (all p > 0.05), but the differences in operative time, stone migration and SFR between the two groups were statistically significant (p < 0.05). Conclusion: This study introduced a new position for ureteroscopic holmium laser lithotripsy for patients with upper ureteral calculi. The Trendelenburg position can improve the SFR and may provide an optional surgical method for treating upper ureteral calculi.

The effect of recombinant and purified tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on the proliferation and apoptosis of PC-3 prostate cancer cells and the 5637 bladder cancer cells were investigated. We used a cell proliferation assay and flow cytometry to measure the proliferation and apoptosis of cancer cells after 24-h incubation of PC-3 and 5637 cells with different concentrations of TRAIL. PC-3 cell proliferation rate significantly decreased when TRAIL was used at concentrations of 20, 40, 80 and 160 ng/ml compared with the untreated group. In the 5637 cells, the proliferation rate significantly decreased when TRAIL was used at concentrations of 5, 10, 20 and 40 ng/ml compared with the untreated group. The flow cytometry results also confirmed that the apoptosis rate of both cancer cell lines increased with TRAIL protein concentration. In conclusion, recombinant and purified TRAIL has anticancer activity by inhibiting proliferation and promoting apoptosis of prostate and bladder cancer cells.

Bladder cancer is one of the malignant tumors that threaten human health. There is an urgent need for a biomarker or treatment target in the treatment of bladder cancer. In humans, enhancer of rudimentary homolog (ERH) is located on chromosome 14q24.1 and is expressed in almost all tissues. Analysis using StarBase v2.0 indicated that ERH expression is up-regulated in urothelial bladder cancer cells (211 cancer samples vs. 19 normal samples). StarBase v2.0 (http://starbase.sysu.edu.cn) starBase Pan-Cancer Analysis Platform is designed for deciphering Pan-Cancer Networks of lncRNAs, miRNAs, ceRNAs and RNA-binding proteins (RBPs) by mining clinical and expression profiles of 14 cancer types (>6000 samples) from The Cancer Genome Atlas (TCGA) Data Portal. Thus, we further explored the impact and mechanisms of ERH knockdown on the proliferation and survival of T24 human bladder cancer cells. The impact of ERH knockdown on cell proliferation, colony formation, and apoptosis was investigated in the T24 cell line using a lentiviral-mediated small interfering RNA (shRNA) strategy. The shRNA strategy efficiently reduced ERH expression, and ERH knockdown impaired cell proliferation, inhibited colony formation, and induced cell apoptosis in T24 human bladder cancer cells. Thus, our study provides evidence for the important role of ERH in the development and progression of human bladder cancer.