Comprehensive analysis of alternative polyadenylation regulators concerning CD276 and immune infiltration in bladder cancer.

Alternative polyadenylation (APA) is emerging as a crucial regulatory mechanism in bladder cancer (BC), while it remains elusive whether APA influences the tumor immune microenvironment (TIME) in BC. We identified two distinct subtypes of BC by APA-related regulatory genes expression profiles. The two subtypes have different pathological grades, prognostic outcomes, tumor immune infiltration characteristics, and pathway enrichment. Subsequently, CPSF3 was identified as a potential immune infiltration-related gene in BC. Highly expressed CPSF3 was positively correlated with unfavorable prognosis and high CD276 expression in BC. Moreover, we verified the expression of CPSF3 in BC tissues and cell lines by qRT-PCR. In conclusion, the study indicates that APA regulatory factors play an important role in immune infiltration of BC, and that CPSF3 was a potentially prognostic marker and immunotherapy target for BC.

Identification of a 5-Gene Signature Predicting Progression and Prognosis of Clear Cell Renal Cell Carcinoma.

BACKGROUND Although the mortality rates of clear cell renal cell carcinoma (ccRCC) have decreased in recent years, the clinical outcome remains highly dependent on the individual patient. Therefore, identifying novel biomarkers for ccRCC patients is crucial. MATERIAL AND METHODS In this study, we obtained RNA sequencing data and clinical information from the TCGA database. Subsequently, we performed integrated bioinformatic analysis that includes differently expressed genes analysis, gene ontology and KEGG pathway analysis, protein-protein interaction analysis, and survival analysis. Moreover, univariate and multivariate Cox proportional hazards regression models were constructed. RESULTS As a result, we identified a total of 263 dysregulated genes that may participate in the metastasis of ccRCC, and established a predictive signature relying on the expression of OTX1, MATN4, PI3, ERVV-2, and NFE4, which could serve as significant progressive and prognostic biomarkers for ccRCC. CONCLUSIONS We identified differentially expressed genes that may be involved in the metastasis of ccRCC. Moreover, a predictive signature based on the expression of OTX1, MATN4, PI3, ERVV-2, and NFE4 could be an independent prognostic factor for ccRCC.

Loss of Fezf2 promotes malignant progression of bladder cancer by regulating the NF-κB signaling pathway.

Forebrain embryonic zinc finger 2 (Fezf2) is an evolutionarily conserved zinc finger transcription repressor. It has been reported to be a tumor suppressor; however, neither the role that Fezf2 plays in bladder cancer nor the mechanisms involved have been investigated. In this study, we showed that Fezf2 expression is downregulated in bladder cancer tissues and cell lines compared to adjacent non-tumor tissues and normal urothelial cells. We also retrospectively analyzed the association between Fezf2 and various clinicopathologic characteristics in 196 bladder cancer patients, and showed that low expression of Fezf2 is correlated with larger tumor size, advanced tumor stage, and poor clinical prognosis. Moreover, we found that overexpression of Fezf2 significantly inhibited the proliferation, growth, migration, and invasion of bladder cancer cells, and attenuated angiogenesis, while knockdown of Fezf2 had the opposite effect. Fezf2 suppressed bladder cancer aggressiveness by activating the NF-κB signaling pathway. These findings suggest that Fezf2 holds promise as a prognostic biomarker, and provide a putative mechanism for bladder cancer progression.

Prognostic Implication of Urothelial Stem Cell Markers Differs According to Primary Tumour Location in Non-Muscle-Invasive Bladder Cancer.

BACKGROUND/AIMS: This study aimed to validate the value of urothelial stem cell (USC) markers ΔNp63, integrin ß4, CD47, and CD44v6 in predicting the prognosis of non-muscle invasive bladder cancer (NMIBC) located in different anatomic regions of bladder. METHODS: The study reviewed the clinicopathologic data of 169 patients with NMIBC. Using real-time PCR and immunohistochemistry, the expression of ΔNp63, integrin ß4, CD47, and CD44v6 in archived specimens of patients with NMIBC were validated. Kaplan-Meier analysis and Cox proportional hazards model were used to assess the prognostic impact of USC markers for recurrent-free survival (RFS). RESULTS: The Real-time PCR data showed that the expression of USC markers were higher in tumors located in the trigone and posterior wall than that in other regions of bladder (P< 0.05). Statistical analysis showed that high expression of ΔNp63 was correlated with tumor stage (P=0.023) and tumor size (P=0.001), that high expression of integrin ß4 was correlated with tumor stage (P=0.026), tumor grade (P=0.005) and tumor size (P=0.003), and that high integrin ß4, CD47, and CD44v6 expression were significantly associated with tumor recurrence (P=0.032, P=0.010, and P=0.043, respectively). Moreover, high expression of ΔNp63 and integrin ß4 was correlated with poor RFS in patients with tumors located in the trigone (P=0.025 and P=0.023, respectively). High expression of integrin ß4, CD47, and CD44v6 was correlated with poor RFS in patients with tumors in the posterior wall (P=0.017, P=0.033 and P=0.047, respectively). High expression of integrin ß4 and CD47 was correlated with poor RFS in patients with tumors in the trigone/posterior wall area (P=0.002 and P=0.005, respectively). CONCLUSION: Our results suggest that USC markers are linked with poor prognosis of NMIBC patients, especially in patients with tumors in the trigone and posterior wall.

Leupaxin Promotes Bladder Cancer Proliferation, Metastasis, and Angiogenesis Through the PI3K/AKT Pathway.

BACKGROUND/AIMS: Leupaxin (LPXN) is a member of the paxillin protein family. Several studies have reported that LPXN regulates cancer development; however, the role of LPXN in bladder cancer remains unknown. METHODS: The expression of LPXN in bladder cancer cells and tissues was determined by real-time PCR, western blotting, and immunohistochemistry, respectively. The biological role of LPXN in bladder cancer cell proliferation, invasion, and angiogenesis was explored both in vitro and in vivo. RESULTS: LPXN expression was elevated in bladder cancer tissues and cell lines compared to adjacent non-tumor tissues and normal urothelial cells. High LPXN expression was correlated with large tumor size, advanced tumor stage, and poor survival in bladder cancer patients. Overexpression of LPXN significantly promoted the proliferation, invasion, and angiogenesis of bladder cancer cells, while suppressing LPXN had the opposite effects. The impact on tumor progression was abolished by inhibiting PI3K/ AKT signaling pathway. We further demonstrated that LPXN probably up-regulated S100P via the PI3K/AKT pathway. CONCLUSIONS: LPXN may facilitate bladder cancer progression by upregulating the expression of S100P via PI3K/AKT pathway. These results provide a novel insight into the role of LPXN in tumorigenesis and progression of bladder cancer and potential therapeutic target of bladder cancer.

HBO1 promotes cell proliferation in bladder cancer via activation of Wnt/ß-catenin signaling.

Histone acetyltransferase binding to ORC1 (HBO1), a histone acetyltransferase, was recently identified as an oncoprotein; however, its role in bladder cancer remains unknown. In this study, we showed that HBO1 was highly expressed at both the mRNA and the protein levels in bladder cancer. HBO1 expression was associated with the clinical features of human bladder cancer, including tumor size (P = 0.018) and T (P = 0.007) classifications. Patients with higher HBO1 expression had shorter recurrence-free survival time, whereas patients with lower HBO1 expression had better survival time. Moreover, we found that ectopic overexpression of HBO1 promoted, whereas HBO1 silencing inhibited tumor growth in bladder cancer cells both in vitro and in vivo. We further demonstrated that upregulation of HBO1 activated the Wnt/ß-catenin signaling pathway and led to nuclear localization of ß-catenin and upregulation of downstream targets of of Wnt/ß-catenin signaling. These findings suggest that HBO1 plays a key role in the progression of bladder cancer via the Wnt/ß-catenin pathway, and may serve as a potential therapeutic target for the treatment of bladder cancer.

Musashi-2 promotes migration and invasion in bladder cancer via activation of the JAK2/STAT3 pathway.

Musashi-2 (Msi2) is considered to have a crucial role in regulating various key cellular functions. However, the clinical significance and biological role of Msi2 in bladder cancer remains unknown. We examined the expression of Msi2 in bladder cancer cell lines in 167 clinical samples and the biological role of Msi2 in bladder cancer cells. Western blotting was used to investigate the possible mechanism of Msi2-induced migration and invasion in bladder cancer. Msi2 was significantly upregulated in bladder cancer cells and tissues compared with normal bladder urothelial cells and tissues. Immunohistochemical analysis revealed high expression of Msi2 in 57 of 167 (34.1%) bladder cancer specimens. Statistical analysis showed a significant correlation of Msi2 expression with advanced clinical stage, lymph node metastasis, and poor prognosis. Overexpression and ablation of Msi2 promoted and inhibited, respectively, the migration and invasion of bladder cancer cells. Furthermore, we found that Msi2 activated the JAK2/STAT3 pathway and promoted expression of genes downstream of JAK2/STAT3 in bladder cancer. This study demonstrates that Msi2 can induce bladder cancer cell migration and invasion by activating the JAK2/STAT3 pathway, and that Msi2 may be a valuable prognostic biomarker for bladder cancer patients.

High expression of constitutive photomorphogenic 1 (COP1) is associated with poor prognosis in bladder cancer.

The present study was to investigate the expression and prognostic value of constitutive photomorphogenic 1 (COP1) in bladder cancer. In our study, real-time quantitative PCR (RT-PCR) was performed to detect 10 pairs of fresh bladder cancer (BCa) and adjacent noncancerous tissues. In addition, immunohistochemistry was utilized to detect the expression of COP1 in 174 clinical bladder cancer samples. What is more, the correlation of COP1 expression and clinicopathological features and clinical outcomes were analyzed. The expression levels of COP1 in clinical bladder cancer were much higher than that in paired adjacent noncancerous tissues (p < 0.0001). High expression of COP1 was closely related with differentiation (p = 0.040) and recurrence (p = 0.001) of patients with bladder cancer. Kaplan-Meier analysis revealed that the expression of COP1 was closely correlated with overall survival (p = 0.048) of bladder cancer, while, recurrence-free survival (p = 0.201). Moreover, Cox multivariate regression analyses showed that COP1 expression was an independent predictor of overall survival (OS; p = 0.027, hazard ratio = 2.127, confidence interval 0.814 to 9.736). Based on our data, the present study suggests that high expression of COP1 may be a novel biological indicator for evaluation of poor prognosis in bladder cancer.

Ultrasound-guided percutaneous intracystic deroofing is effective in the treatment of simple renal cysts.

OBJECTIVE: To investigate the efficacy and safety of ultrasound-guided percutaneous intracystic deroofing for the treatment of simple renal cysts. METHODS: A retrospective study was conducted to analyze the clinical data of 46 patients with dorsal exophytic simple renal cysts treated at the First Affiliated Hospital of Nanchang University between February 2017 and June 2022. The patients were divided into two groups according to the surgical method, with 20 cases undergoing ultrasound-guided percutaneous intracystic deroofing being assigned to the observation group and 26 cases treated by retroperitoneal laparoscopic renal cyst removal included in the control group. The operation time, blood loss, postoperative catheterization time, postoperative drainage tube indwelling time, postoperative hospital stay, and complications were compared. RESULTS: None of the 46 patients converted to open surgery. The observation group showed significantly less blood loss, shorter operation time, drainage tube drainage time, postoperative hospital stay, and indwelling catheter time than the control group (all P<0.05). The two procedures had a success rate of 100%. There were no statistical significances in K+, Na+, or serum creatinine between the two groups (all P>0.05). All patients were followed up (3 to 6 months) after surgery, and no cyst recurrence was found by imaging examination. CONCLUSIONS: Ultrasound-guided percutaneous intracystic deroofing of renal cysts is worthy of clinical application in the treatment of simple renal cysts due to its significant advantages such as short operation time, less trauma, quick recovery, safety, effectiveness, and low cost.

Mechanisms of prostate atrophy after LHRH antagonist cetrorelix injection: an experimental study in a rat model of benign prostatic hyperplasia.

In the present study, we investigated the roles of TGF-ß signaling pathway in a rat benign prostatic hyperplasia (BPH) model treated with cetrorelix. TGF-ß1 and c-Myc expression were measured by qRT-PCR and Western blotting in the proximal and distal region of ventral prostatic lobes, respectively. We observed that treatment with cetrorelix led to a significant reduction of ventral prostate weight in a dose-dependent manner. In the proximal region, after cetrorelix treatment, the expression of TGF-ß1 was dramatically increased (P<0.05), while the expression of c-Myc was significantly decreased (P<0.05). In comparison with the control group, the cetrorelix groups had more TUNEL-positive cells. Our findings strongly suggest that the TGF-ß signaling pathway may be one of the major causes responsible for prostate volume reduction in BPH rats after cetrorelix treatment.

Experience in management of Fournier's gangrene: a report of 24 cases.

Fournier's gangrene (FG) is an extremely aggressive and rapidly progressive polymicrobial soft tissue infection of the perineum, anal area or genitalial regions with a high mortality rate. The objectives of this study were to share our experience with the management of this serious infectious disease over the last 15 years. This retrospective study examined 24 patients diagnosed as having FG who were admitted to our hospital between March 1996 and December 2011. The gender, age, etiology, predisposing factors, laboratory findings, treatment modality, hospitalization time and spread of gangrene of the subjects were all recorded and analyzed. The results showed that the mean age of the patients was 48.33 years, the male-to-female ratio was 5:1 and the mortality rate was 20.8% (5/24). The most common predisposing factor was diabetes mellitus in 10 patients (41.6%), followed by alcohol abuse, obesity, neoplasms and immunosuppression. The most common etiology was peri-anal and peri-rectal abscesses (45.8%), followed by lesions of urogenital origin (33.3%) and cutaneous (8.3%) origin. No local pathologies could be identified in 3 (12.5%) patients. The most commonly isolated microorganisms were Escherichia coli (62.5%), followed by Enterococcus, Pseudomonas aeruginosa and Staphylococcus aureus. The median admission Fournier's gangrene severity index (FGSI) score for survivors was 5.63±1.89 against 13.6±3.64 for non-survivors which was designed for predicting the disease severity in the series. Early diagnosis and immediate extensive surgical debridement were significant prognostic factors in the management of Fournier gangrene. Individualized reconstructive modalities for wound coverage were useful in that they repaired the tissue defect and improved the quality of life. We are led to conclude that Fournier's gangrene is a severe condition with a high mortality. The Fournier's gangrene severity index (FGSI) score at admission serves as a good predictor for the disease severity. Early diagnosis, surgical debridement and aggressive fluid therapy are significant prognostic factors in the management of Fournier gangrene. Individualized reconstructive surgery modalities for wound coverage are useful to correct the tissue defect and improve the quality of life.

Standard versus mini-percutaneous nephrolithotomy for renal stones: a meta-analysis.

OBJECTIVES: To compare the standard percutaneous nephrolithotomy and mini-percutaneous nephrolithotomy in order to determine the optimal tract size for patients with renal stones. METHODS: A systematic search of Web of Science, EMBASE, Cochrane Library, and PubMed databases was conducted for articles published through 20 August 2019, reporting on a comparison of the standard percutaneous nephrolithotomy and mini-percutaneous nephrolithotomy using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. RESULTS: Of 763 studies, 14 were considered for the evidence synthesis. A total of 1980 cases were included. Of these patients, 897 cases underwent standard percutaneous nephrolithotomy, and 1083 cases underwent mini-percutaneous nephrolithotomy. Stone-free rates were 87.6% (786 of 897 patients) for standard percutaneous nephrolithotomy and 87.8% (951 of 1083 patients) for mini-percutaneous nephrolithotomy (p = 0.57). Tract sizes of 30F and 22-26F in standard percutaneous nephrolithotomy group shorten operation time compared with mini-percutaneous nephrolithotomy (p = 0.02; p = 0.004; respectively). Leakage (p = 0.04), bleeding (p = 0.01), blood transfusion (p < 0.00001), and renal pelvis perforation (p = 0.02) were more common in standard percutaneous nephrolithotomy group than in mini-percutaneous nephrolithotomy group. Subgroup analysis showed only blood transfusion for 30F and 22-26F standard percutaneous nephrolithotomy group was more common than mini-percutaneous nephrolithotomy (p < 0.0001, p = 0.005, respectively). CONCLUSIONS: Standard percutaneous nephrolithotomy was associated with higher leakage, bleeding, blood transfusion, and renal pelvis perforation, but had a shorter operation time. Tract size of 30F improved the stone-free rate compared with mini-percutaneous nephrolithotomy, but led to more complications. Tract size of 22-26F was no better than 30F or mini-percutaneous nephrolithotomy.

Overexpression of LINC00160 predicts poor outcome and promotes progression of clear cell renal cell carcinoma.

Clear cell renal cell carcinoma (ccRCC) is the most common subtype of renal carcinoma and exhibits a high risk of invasion and metastasis. It is urgent to uncover a novel biomarker and clarify the underlying mechanism for ccRCC progression and metastasis. Although accumulating research has demonstrated that long non-coding RNAs (lncRNAs) play crucial roles in tumor progression, numerous lncRNAs in ccRCC are largely unknown. Therefore, we screened the differentially expressed lncRNAs among several GEO datasets and chose LNC00160 for further investigation. LNC00160 was significantly upregulated in ccRCC and high expression predicted poor prognosis; higher expression of LNC00160 was associated with advanced clinic pathological parameters in TCGA_KIRC Cohort. Knockdown of LNC00160 suppressed malignancy of ccRCC in vitro and in vivo. Correlation analysis and gene set enrichment analysis (GSEA) revealed that LNC00160 might be associated with Wnt signaling pathway, mTOR signaling pathway, fatty acid metabolism and cell cycle. In conclusion, our results demonstrated that LNC00160 acted as an oncogenic gene and a specific prognostic indicator for patients with ccRCC, and that LNC00160 might be a targeted intervention for ccRCC patients in the future.

The immune infiltration in clear cell Renal Cell Carcinoma and their clinical implications: A study based on TCGA and GEO databases.

The tumor immune microenvironment in clear cell Renal Cell Carcinoma (ccRCC) still remains poorly understood. Previous methods to study the tumor immune microenvironment have a limitation when accounting for the functionally distinct cell types. In this study, we investigated the differently infiltrated immune cells and their clinical significance in ccRCC for the purpose of shedding some important light on the complex immune microenvironment in ccRCC. The devolution algorithm (CIBERSORT) was applied to infer the proportion of 22 immune infiltrating cells based on gene expression profiles of ccRCC bulk tissue, which were downloaded from TCGA and GEO databases. As a result, we observed considerable differences in immune cells percentage between ccRCC tumor tissue and paired normal tissue; meanwhile, we uncovered their internal correlations and associations with Fuhrman grade. Moreover, dendritic cells resting, dendritic cells activated, mast cells resting, mast cells activated and eosinophils were associated with favorable prognosis, whereas B cells memory, T cells follicular helper and T cells regulatory (Tregs) were correlated with poorer outcome.

A cluster of metabolism-related genes predict prognosis and progression of clear cell renal cell carcinoma.

Clear cell renal cell carcinoma (ccRCC) has long been considered as a metabolic disease characterized by metabolic reprogramming due to the abnormal accumulation of lipid droplets in the cytoplasm. However, the prognostic value of metabolism-related genes in ccRCC remains unclear. In our study, we investigated the associations between metabolism-related gene profile and prognosis of ccRCC patients in the Cancer Genome Atlas (TCGA) database. Importantly, we first constructed a metabolism-related prognostic model based on ten genes (ALDH6A1, FBP1, HAO2, TYMP, PSAT1, IL4I1, P4HA3, HK3, CPT1B, and CYP26A1) using Lasso cox regression analysis. The Kaplan-Meier analysis revealed that our model efficiently predicts prognosis in TCGA_KIRC Cohort and the clinical proteomic tumor analysis consortium (CPTAC_ccRCC) Cohort. Using time-dependent ROC analysis, we showed the model has optimal performance in predicting long-term survival. Besides, the multivariate Cox regression analysis demonstrated our model is an independent prognostic factor. The risk score calculated for each patient was significantly associated with various clinicopathological parameters. Notably, the gene set enrichment analysis indicated that fatty acid metabolism was enriched considerably in low-risk patients. In contrast, the high-risk patients were more associated with non-metabolic pathways. In summary, our study provides novel insight into metabolism-related genes' roles in ccRCC.

Correction: CLCA4 inhibits bladder cancer cell proliferation, migration, and invasion by suppressing the PI3K/AKT pathway.

[This corrects the article DOI: 10.18632/oncotarget.21724.].

CLCA4 inhibits bladder cancer cell proliferation, migration, and invasion by suppressing the PI3K/AKT pathway.

Calcium activated chloride channel A4 (CLCA4), a tumor suppressor, was shown to contribute to the progression of several human cancers, while its role in bladder carcinoma remains unclear. In this study, we showed CLCA4 expression was down-regulated in bladder carcinoma tissues and cells compared to adjacent non-tumor tissues and normal urothelial cells. Low CLCA4 expression was correlated with larger tumor size, advanced tumor stage, and poor prognosis in bladder carcinoma patients. Overexpression of CLCA4 profoundly attenuated the proliferation, growth, migratory and invasive capabilities of bladder cancer cells, whereas CLCA4 knockdown had the opposite effect. Mechanistically, CLCA4 is involved in PI3K/AKT signaling and its downstream molecules can promote bladder cancer cell proliferation. Additionally, CLCA4 could mediate the migration and invasion of bladder cancer cells by regulating epithelial-mesenchymal transition and PI3K/Akt activation. This study suggests that CLCA4 may represent a promising prognostic biomarker for bladder cancer and provides a possible mechanism for bladder cancer growth and invasion.

Inhibition of NEDD4 inhibits cell growth and invasion and induces cell apoptosis in bladder cancer cells.

The neural precursor cell expressed developmentally downregulated protein 4 (NEDD4) plays a pivotal oncogenic role in various types of human cancers. However, the function of NEDD4 in bladder cancer has not been fully investigated. In the present study, we aim to explore whether NEDD4 governs cell proliferation, apoptosis, migration, and invasion in bladder cancer cells. Our results showed that downregulation of NEDD4 suppressed cell proliferation in bladder cancer cells. Moreover, we found that inhibition of NEDD4 significantly induced cell apoptosis. Furthermore, downregulation of NEDD4 retarded cell migration and invasion. Notably, overexpression of NEDD4 enhanced cell growth and inhibited apoptosis. Consistently, upregulation of NEDD4 promoted cell migration and invasion in bladder cancer cells. Mechanically, our Western blotting results revealed that downregulation of NEDD4 activated PTEN and inhibited Notch-1 expression, whereas upregulation of NEDD4 reduced PTEN level and increased Notch-1 level in bladder cancer cells. Our findings indicated that NEDD4 exerts its oncogenic function partly due to regulation of PTEN and Notch-1 in bladder cancer cells. These results further revealed that targeting NEDD4 could be a useful approach for the treatment of bladder cancer.

Association of increased urine brain derived neurotrophic factor with lower urinary tract symptoms in men with benign prostatic hyperplasia.

Urinary brain-derived neurotrophic factor (BDNF), an ubiquitous neurotrophin, was found to rise in patients with benign prostatic hyperplasia (BPH). We hypothesized that the urinary level of BDNF could be a potential biomarker for lower urinary tract symptoms (LUTS) in patients with BPH. Totally, 76 patients with BPH-caused LUTS and 32 male control subjects without BPH were enrolled. International Prostate Symptom Score (IPSS) was applied to assess the symptom severity of LUTS. Urodynamic tests were performed for the diagnosis of underlying detrusor overactivity (DO) in the patients with BPH. Urine samples were collected from all subjects. Urinary BDNF levels were measured using enzyme-linked immunosorbent assays and normalized by urinary creatinine (Cr) levels. Seventy-six BPH patients were divided into moderate LUTS group (n=51, 720) according to the IPSS. Of the 76 BPH patients, DO was present in 34 (44.7%) according to the urodynamic test. The urinary BDNF/Cr levels were significantly higher in BPH patients with moderate LUTS (8.29±3.635, P<0.0001) and severe LUTS (11.8±6.44, P<0.0001) than normal controls (1.71±0.555). Patients with severe LUTS tended to have higher urinary BDNF/Cr levels than patients with moderate LUTS (11.8±6.44 vs. 8.29±3.635, P=0.000). The conditions of BPH with LUTS correlated with elevated urinary BDNF levels, and urinary BDNF levels were even higher in BPH-DO patients. The results of this study have provided evidence to suggest that urinary BDNF level test could evaluate the severity of LUTS in BPH patients, and BDNF level can be used as a biomarker for the diagnosis of DO in BPH patients.

Combining Protein and miRNA Quantification for Bladder Cancer Analysis.

We combine the telomerase extension reaction and microRNA (miRNA)-induced rolling circle amplification, followed by graphene oxide (GO) and nicking enzyme-assisted signal amplification as a method to analyze telomerase and miRNA-21 in urine samples with the following merits. First, it is a binary assay and can simultaneously output double signals that correspond to the quantities of telomerase and miRNA, respectively. Second, telomerase activity is enhanced by using a DNA molecular beacon probe to inhibit the formation of G-quadruplex. Third, background noise is decreased significantly via introduction of GO. Fourth, performance tests on about 258 urine samples demonstrate that this binary assay can distinguish between urine from bladder cancer patients, those with cystitis, and normal individuals. Finally, this strategy also shows great potential in distinguishing between muscle-invasive bladder cancers and non-muscle-invasive bladder cancers. The proposed strategy will greatly contribute to clinical decision-making and individualized treatments.