The LMR-SSIGN-MAPS model predicts disease-free survival in patients with localized clear cell renal cell carcinoma.

Introduction: Prediction models are increasingly being used to predict outcomes after surgery, and such a model would be a precious tool for patients with clear cell renal cell carcinoma (ccRCC) after surgery. Aim: To develop a comprehensive model for predicting disease-free survival (DFS) in patients with localized ccRCC. Material and methods: In a retrospective analysis of 612 patients, least absolute shrinkage and selection operator (LASSO) Cox regression analysis was performed to identify significant predictors, and then risk factors were used to construct a prognostic model. Harrell's concordance index (C-index) was used to assess the accuracy of the model. Results: The lymphocyte-to-monocyte ratio (LMR), Mayo Clinic stage, size, grade, necrosis score (SSIGN), and Mayo adhesive probability score (MAPS) were the significant risk factors screened by LASSO Cox regression and reconfirmed by multivariate Cox regression analysis in 44 variables. Then a model was constructed by combining the LMR, SSIGN, and MAPS. The C-index of the LMR-SSIGN-MAPS model was greater than the SSIGN score alone. Kaplan-Meier survival analysis demonstrated a significant association between higher LMR-SSIGN-MAPS score and poorer DFS. Conclusions: The LMR-SSIGN-MAPS model, which consists of preoperative inflammation biomarkers, a perinephric adipose tissue image-based scoring system, and pathological features, showed the strengths of easy-to-use and high predictability and might also be used as a promising prognosis model in predicting DFS for patients with localized ccRCC.

Comparative risk of acute kidney injury among cancer patients treated with immune checkpoint inhibitors.

With the development and introduction of immune checkpoint inhibitors (ICIs) in cancer patients, immune-related side effects have increasingly attracted attention. However, the risks of immune-related renal toxicity are poorly characterized. In this study, we performed a network meta-analysis (NMA) of ICI-related randomized clinical trials (RCTs) to elucidate the comparative risk of acute kidney injury (AKI) in cancer patients receiving different ICIs. We also sought to identify other factors potentially affecting the risk of AKI. PubMed and EMBASE were searched for peer-reviewed trial reports published between January 2000 and May 2021. Eligible studies were RCTs studying ICIs in cancer patients and reporting AKI data. We performed a frequentist NMA to evaluate the risk ratios for grade 1-5 and grade 3-5 AKI between the treatment groups. We also assessed the absolute incidence of AKI in the ICI-containing arm using traditional direct meta-analysis. Once significant heterogeneity was detected in a traditional direct meta-analysis, multivariable meta-regression analysis was applied to identify factors that significantly affected the absolute incidence of AKI. A total of 85 RCTs were included in this study. In the NMA for the risk of grade 1-5 and 3-5 AKI, ipilimumab showed a significantly higher risk than avelumab and durvalumab, whereas 1 mg/kg nivolumab plus 3 mg/kg ipilimumab (N1I3) showed a significantly higher risk than other groups. In terms of treatment ranking, durvalumab ± low-dose tremelimumab and avelumab were consistently among the top three safest treatments for grade 1-5 or 3-5 AKI, whereas N1I3, ipilimumab and tremelimumab were consistently among the top three treatments with the highest risk for grade 1-5 or 3-5 AKI. Compared with other cancers, renal cell carcinoma and urothelial carcinoma showed a significantly higher risk of AKI. The incidence of AKI was significantly higher with ICI+chemotherapy than with ICI monotherapy. In this NMA involving large-scale up-to-date ICI trials, we demonstrated the comparative safety of existing ICI drugs for grade 1-5 and grade 3-5 AKI. Based on data from the ICI arms of these trials, we also revealed several potential risk factors for immune-related AKI, including tumor type and treatment paradigm.

Urine PD-L1 is a tumor tissue candidate substitute and is associated with poor survival in muscle-invasive bladder cancer patients.

Programmed death molecule ligand 1 (PD-L1) expression in urothelial carcinoma is a predictive marker used to guide immunotherapy. As expression of PD-L1 may be heterogeneous in the tumor tissue space, it cannot be accurately determined by immunohistochemical analysis. In this study, we examined PD-L1 protein levels in preoperative urine samples from bladder cancer patients, evaluated the prevalence of PD-L1 in urine, examined the usefulness of urine as a surrogate for PD-L1 expression in tumors, and compared PD-L1 expression in postoperative pathological sections. We found that PD-L1 in urine and tumor tissue correlated well and that it may be able to some extent serve as a surrogate for tissues in bladder cancer and thus predict risk of recurrence in muscle-invasive bladder cancer (MIBC) patients. Our findings reveal the clinical relevance of urine PD-L1 as a noninvasive prognostic indicator for immunotherapy and offer clinical translational suggestions for eventual development of a prognostic model for immunotherapy for bladder cancer.

The effect of enhanced recovery after surgery protocols combined with laparoscopic radical cystectomy with intracorporeal urinary diversion / 中华泌尿外科杂志

Objective:To explore the effect of enhanced recovery after surgery (ERAS) protocols in patients undergoing laparoscopic radical cystectomy (LRC) and intracorporeal urinary diversion (ICUD).Methods:A total of 83 patients who received LRC+ ICUD in Beijing Chaoyang Hospital from March 2014 to September 2020, were divided into 2 groups based on different perioperative management, including 29 ERAS cases and 54 conventional recovery after surgery (CRAS) cases. The ERAS group included 26 males and 3 females , with an average age of (62.07 ± 9.26) years. There were 26 patients with ASA class Ⅰ-Ⅱ, 3 patients with ASA class Ⅲ, 4 patients received neoadjuvant chemotherapy, and 7 patients had a history of abdominal surgery in ERAS group. The CRAS group included 44 males and 10 females , with an average age of (61.59 ± 10.16) years. There were 50 patients with ASA class Ⅰ-Ⅱ, 4 patients with ASA class Ⅲ, 9 patients received neoadjuvant chemotherapy, and 10 patients had a history of abdominal surgery in CRAS group. There were no statistically significant differences in the baseline characteristics between the two groups. The patients in both groups underwent LRC+ ICUD procedures. The perioperative results and complications between the two groups were compared.Results:In the ERAS group, there were 20 patients who underwent Bricker ileal conduit surgery and 9 patients who underwent Studer orthotopic ileal neobladder surgery. Pathological staging included 3, 3, 7, 7, 5 and 4 cases in stage T a, T is, T 1, T 2, T 3 and T 4a, respectively. There were 23, 2, 3 and 1 patient with pathological stage N 0, N 1, N 2 and N 3, respectively. Pathological diagnosis included 3 cases of low-grade urothelial carcinoma, 24 cases of high-grade urothelial carcinoma, and 2 cases of other histological subtypes. In the CRAS group, there were 31 patients who underwent Bricker ileal conduit surgery and 23 patients who underwent Studer orthotopic ileal neobladder surgery. Pathological staging included 5, 3, 12, 9, 15 and 10 patients in stage T a, T is, T 1, T 2, T 3 and T 4a, respectively. There were 35, 6, 7 and 6 patients with pathological stage N 0, N 1, N 2, and N 3, respectively. Pathological diagnoses included 6 cases of low-grade urothelial carcinoma, 45 cases of high-grade urothelial carcinoma, and 3 cases of other histological subtypes. There were no statistically significant differences ( P>0.05) in surgical methods, pathological staging, or pathological types between the ERAS and CRAS groups. ERAS group presented less albumin loss [(25.73±8.63)% vs. (32.63±9.05)%, P=0.001], shorter hospital stay [9(7, 13)d vs. 12(9, 16)d, P=0.006], less 30-day overall complications [55.2% (16/29) vs. 83.3% (45/54), P=0.009]. In multivariable analysis, maximum albumin loss≥20% was independently associated with 30-day minor complications ( P=0.049), and maximum albumin loss ≥25% was independently associated with hospital of stay≥10 days ( P=0.038), respectively. Conclusions:For patients who received LRC+ ICUD, ERAS was associated with reduced perioperative albumin loss, shorter length of stay, less 30-day complications, accelerated recovery time, improved clinical outcome and less albumin injection.

Effects of diabetes and obesity on renal function recovery in patients under going laparoscopic partial nephrectomy / 现代泌尿外科杂志

【Objective】 To investigate the effects of diabetes and/or obesity on the incidence of acute kidney injury (AKI) and long-term renal function recovery in patients undergoing laparoscopic partial nephrectomy. 【Methods】 A retrospective analysis was performed on 221 patients who underwent laparoscopic partial nephrectomy in Beijing Chaoyang Hospital during Jan.2018 and Dec.2019. Baseline data, incidence of AKI, and renal function recovery after 12 months were analyzed in the diabetic, non-diabetic, diabetic + obese and diabetic + non-obese groups, with estimated glomerular filtration rate (eGFR) reaching 90% of the preoperative level as the standard. Risk factors of AKI were analyzed with univariate and multivariate regression analyses. 【Results】 Compared with the non-diabetic group, the diabetic group had lower preoperative eGFR [ (79.1±12.1)mL/ (min·1.73 m2 )vs. (85.3±10.7)mL/ (min·1.73 m2 ), P=0.01] , higher incidence of AKI (14.0% vs. 11.8%), and lower proportion of patients whose renal function recovered to at least 90% of the preoperative level (73.1% vs.83.5%). Patients with diabetes and obesity were more likely to develop AKI (31.3% vs.3.7%, P=0.039), and diabetic patients without obesity had better renal function recovery (81.3% vs. 60.0%). Multivariate analysis showed gender and operation time were the independent risk factors of AKI. 【Conclusion】 There were no significant differences between diabetic and non-diabetic patients in the incidence of postoperative AKI, or rate of renal function recovery 12 months after surgery. The incidence of AKI significantly increased in diabetic patients with obesity, and the renal function recovery rate decreased one year after surgery. Gender and operation time were the independent risk factors of AKI.

Biotransformation of Abiraterone Into Five Characteristic Metabolites by the Rat Gut Microbiota and Liver Microsomes.

It is well known that the role of gut microbiota in drug metabolism, especially in oral difficult absorbable drugs. Understanding the gut microbiota could enable us to understand drugs in new ways. The purpose of the study was to investigate explore the metabolites of the anti-prostate cancer drug Abiraterone by examining gut microbiota metabolism and hepatic metabolism in vitro. In this study, five metabolites (M1, M2, M3, M4 and M5) of Abiraterone were discovered using LC/MSn-IT-TOF. Four isomeric metabolites M1-M4 were found in liver microsome. M5 was found in the intestinal contents of Sprague-Dawley rats with a molecular weight of 388.31. Among them, M4 was found to be Abiraterone N-Oxide by comparison with the standard sample. After further comparing the metabolic behavior of Abiraterone in rat gut microbiota and liver microsomes, we delineated the possible metabolic pathways of Abiraterone. In conclusion, Abiraterone is metabolized specifically in liver microsomes and gut microbiota. This study can provide a theoretical basis for elucidating the metabolic mechanism of Abiraterone and guide its rational application in clinic.

Feasibility of single position laparoscopic radical nephrectomy and tumor thrombectomy for left renal cell carcinoma with high-risk Mayo grade 0 and 1 tumor thrombus.

BACKGROUND: To explore the feasibility of single-position laparoscopic radical nephrectomy (LRN) and tumor thrombectomy for left renal cell carcinoma with high-risk Mayo 0 and 1 tumor thrombus (TT). METHODS: All patients with left renal cell carcinoma and venous TT (high-risk Mayo grade 0 and 1) who were performed single-position LRN and tumor thrombectomy were involved. After the renal artery was controlled by Hem-o-lok, the left renal vein was dissected through descending colon mesentery. The left renal vein was divided by EndoGIA for high-risk Mayo grade 0 TT. For Mayo grade 1 TT, part of the inferior vena cava was blocked by a bulldog clamp after milking the TT into the left renal vein and the inferior vena cava was sutured after complete excision of the TT. RESULTS: 3 patients were involved and operations were performed successfully without conversion to open surgery. The mean operation time was 136 min and the mean estimated blood loss was 60 mL. No postoperative complications occurred. CONCLUSIONS: It is feasible to control left renal vein and partial inferior vena cava through descending colon mesentery in a single position during LRN and tumor thrombectomy for the treatment of high-risk Mayo grade 0 and 1 TT.

Detection of PD-L1 in the urine of patients with urothelial carcinoma of the bladder.

There are currently five programmed death-1 (PD-1)/programmed death ligand-1 (PD-L1) inhibitors approved for the treatment of locally advanced or metastatic urothelial carcinoma (UC) of the bladder. For platinum-ineligible patients, testing of tumor specimens for PD-L1 expression is required. However, scoring of PD-L1 immunohistochemistry is complex due to different antibodies used, the requirement to score expression in different cellular compartments and intratumoral heterogeneity. It can also be difficult to obtain and test longitudinal tumor samples, which would be desirable to monitor treatment responses and tumor evolution under treatment-induced selective pressure. In the present proof-of concept study, we provide evidence that PD-L1 can be detected in the urine of patients with non-muscle invasive bladder cancer (NMIBC) and muscle-invasive bladder cancer (MIBC). Urine PD-L1 levels were significantly higher in NMIBC and MIBC patients when compared to patients with various non-malignant urological diseases. Further prospective and independent studies are required to assess the value of PD-L1 in the urine as a novel biomarker with potential for the early detection, prediction and therapeutic monitoring of patients with UC of the bladder.

The learning effect of modified ileal conduit intracorporeally accomplished following laparoscopic radical cystectomy / 中华泌尿外科杂志

Objective:To analyze the learning effect of laparoscopic radical cystectomy(LRC)+ modified ileal conduit(MIC).Methods:From 2014 to 2019, 42 patients underwent MIC and their clinical data was retrospectively analyzed. 34 operations were performed by surgeon 1 and 8 operations by surgeon 2. We divided the 34 patients of surgeon 1 into three groups according to their surgical sequence (group A, 1st to 12th; group B, 13th to 23th; group C, 24 th to 34 th), the 8 cases of surgeon 2 was regarded as group D. The history of abdomen surgery in the 4 groups were 0, 1, 4, 3 cases, respectively ( P<0.05). There was no significant difference of the other baseline characteristics, such as age, BMI, American Society of Anesthesiologists. Then we compared several variables between the 4 groups like operation time, time of ileal conduit construction, blood loss, complication rate, lymph node yield, surgical margin, etc. The key steps of the MIC included isolating terminal ileum when the mesentery was transilluminated, performing end-to-end reflux ureterointestinal anastomosis after the efferent loop was fixed, closing the rent of the retroperitoneum. Results:All operations were performed intracorporeally with no transition to open surgery. The operative time for group A, B, C were 330.0(320.0, 360.0)min, 300.0(250.0, 308.0)min, 270.0(216.0, 324.0)min, respectively ( P =0.010). The time of ileal conduit construction of the 3 groups were 136.5(131.3, 147.5)min, 92.0(79.0, 119.0)min, 79.0(72.0, 115.0)min, respectively ( P <0.001). In addition, the difference of the two variables above between A and B, A and C groups separately reached statistical significance ( P<0.05), while the difference between B and C groups did not ( P>0.05). Other variables, such as blood loss, complication rate, lymph node yield, surgical margin, between the 3 groups reached no statistical significance ( P>0.05). The operative time of group D was 420.0(350.0, 450.0)min, and it reached statistical significance ( P<0.05) when compared with group A. There were no significant differences in other variables, such as blood loss, complication rate, lymph node yield, surgical margin, among the 2 groups ( P>0.05). Conclusions:The learning effect of LRC+ MIC was obvious. When surgeon volume increased, the operative time decreased significantly. Variables like estimated blood loss and complication rate of the 2 surgeons did not reached significant difference, which indicated reproductivity and safety of this procedure.

Evaluation of the early value of 68Ga-PSMA PET/CT for radical prostatectomy / 中华泌尿外科杂志

Objective:To evaluate the value of 68Ga-labeled prostate-specific membrane antigen( 68Ga-PSMA PET/CT)in the diagnosis of prostate cancer(PCa) and determination of the strategy on neurovascular bundle(NVB)preservation and lymphadenectomy before surgery. Methods:We retrospectively analyzed the clinical data of 46 patients with newly diagnosed PCa who underwent 68Ga-PSMA PET/CT from June 2018 to October 2019. The median age was 66.50 (60.00, 69.25) years old and the median PSA was 15.97(8.58, 33.10)ng/ml. Forty-one patients were diagnosed PCa and 5 were diagnosed with benign prostatic hyperplasia or prostatitis by 68Ga-PSMA PET/CT, 6 were diagnosed with lymph nodes metastasis. Forty patients underwent mpMRI, 33 were diagnosed PCa, and 6 were diagnosed with lymph nodes metastasis. Seventeen patients underwent 11C-choline PET/CT, 12 were diagnosed PCa, and 4 were diagnosed with lymph nodes metastasis. Among the 41 patients which were diagnosed PCa by 68Ga-PSMA PET/CT, 26 were confirmed with PCa by needling biopsy, 12 did not undergo the needling biopsy and 3 had negative biopsy; 22 were in the high-risk group, and 19 were in the low- and medium-risk group. All 41 patients underwent radical prostatectomy. The strategy of NVB preservation was determined by the position of the PCa reported by 68Ga-PSMA PET/CT. If the tumor was close to the unilateral prostate capsule, the healthy part of the NVB would be preserved. And if the tumor was limited in the prostate gland, bilateral NVB would be preserved. The NVB was preserved in 16 cases(6 cases unilateral and 10 cases bilateral). In addition to routine lymph node dissection for patients in the high-risk group, lymph node dissection was also performed on patients with 68Ga-PSMA PET/CT that showed positive lymph nodes in the low- and medium-risk groups. The paired chi-square or Fisher exact test was used to compare the sensitivity and specificity of 68Ga-PSMA PET/CT, mpMRI, and 11C-choline PET/CT for lesion detection. Spearman analysis was used to examine the correlation between the SUV max, Gleason score, and the PSA value before treatment. Results:Forty-one patients undergoing radical resection were diagnosed PCa by postoperative pathology, and no cancer tissue was observed at the surgical margin; the median Gleason score was 8(7, 9); 20 cases (48.8%) had a pathological stage ≤pT 2c; 21 cases (51.2%)≥pT 3; 7 cases were N+ (11 positive lymph nodes). Seven cases (17.1%) had complications of Clavien-Dindo ≤ grade 2 within 30 days after surgery, and there was no complication above grade 3 after surgery. The median follow-up time of the 41 patients was 16(12, 20). The rate of urinary control was 46.3%, 95.1%, and 100% after 1 month, 6 months, and 12 months follow-up respectively. Among the five patients that did not undergo surgery, the PSA value of 4 decreased after antibiotic treatment, and biopsy was performed in 1 case without PSA decreasing, and no carcinoma was found. The sensitivity of 68Ga-PSMA PET/CT on the primary PCa was 100%(41/41), which was significantly better than that of 11C-choline(80%, 12/15, P=0.016)and mpMRI (83.7%, 31/37, P=0.009), while no statistical significance was observed between the specificity of 68Ga-PSMA PET/CT (5/5)and 11C-choline PET/CT(2/2, P=1.000), 68Ga-PSMA PET/CT and mpMRI(1/3, P=0.107). Of the 41 patients that were diagnosed with PCa, the sensitivity of 68Ga-PSMA PET/CT to lymph nodes metastases(71.4%, 5/7) had a significant difference with that of mpMRI(16.7, 1/6, P=0.016), but no statistical difference with 11C-choline (75%, 3/4, P=1.000). Analysis of the relationship between SUV max of 68Ga-PSMA PET/CT, Gleason score, and PSA value before treatment revealed that the SUV max of 68Ga-PSMA PET/CT in patients with Gleason score ≥8 and <8 score were 19.60(9.58, 24.38) and 8.55 (5.18, 12.88); SUV max of patients with PSA values ≥20 ng/ml and <20 ng/ml before treatment were 19.40 (13.00, 23.50) and 8.40 (5.35, 13.95), respectively, the differences were statistically significant (all P<0.05). Conclusions:68Ga-PSMA PET/CT had high sensitivity and specificity for the diagnosis of primary prostate cancer lesions, but the sensitivity for the diagnosis of lymph node metastasis was not enough to guide the preoperative decision of whether to remove the lymph node and the scope of the removal. However, the treatment strategy of whether to retain NVB could be formulated according to the tumor location displayed by 68Ga-PSMA PET/CT before surgery.

MLL5α activates AR/NDRG1 signaling to suppress prostate cancer progression.

Prostate cancer (PCa) is one of the most prevalent malignancies in men. However, the molecular mechanism controlling the transformation of androgen-dependent PCa (ADPC) to castration-resistant PCa (CRPC) is largely unknown. Androgen receptor (AR) signaling has been reported to play a key role in this process; thus, searching for the novel AR co-activator is important for identifying the mechanism underlying PCa progression. In this study, we focused on the function of mixed lineage leukemia-5α (MLL5α), an epigenetic regulator that exhibits aberrant expression in PCa. MLL5α was the primary expressed form of MLL5 protein in PCa cells and it significantly suppressed proliferation, invasion, and migration in PCa cell lines. Upon stimulation with dihydrotestosterone (DHT), knockdown of MLL5α significantly suppressed N-myc downstream regulated gene 1 (NDRG1) and Kallikrein-related peptidase 3 (KLK3) expression. MLL5α directly bound with AR on the androgen response elements (AREs) and recruited H3K4me3 to the promoters of NDRG1 and KLK3. Downregulation of NDRG1 partially restored the cell invasion and migration suppressed by MLL5α. As evaluated by the proliferation of PCa cells, overexpression of MLL5α synergistically promoted sensitivity to enzalutamide (ENZ) treatment. In PCa patients, MLL5α expression was lower in the high Gleason score (GS) (GS > 7) group than in the low GS (GS < 7) group. In conclusion, suppression of AR/NDRG1 signaling via androgen deprivation therapy (ADT) may be a potential mechanism of CRPC progression. MLL5α significantly suppressed PCa progression by promoting AR/NDRG1 signaling, indicating that regulating MLL5α expression may be a potential treatment approach for patients with advanced PCa.

The long-term efficacy of one-shot neoadjuvant intra-arterial chemotherapy combined with radical cystectomy versus radical cystectomy alone for bladder cancer: a propensity-score matching study.

BACKGROUND: Bladder cancer is a complex disease associated with high morbidity and mortality. Management of bladder cancer before radical cystectomy continues to be controversial. We compared the long-term efficacy of one-shot neoadjuvant intra-arterial chemotherapy (IAC) versus no IAC (NIAC) before radical cystectomy (RC) for bladder cancer. METHODS: We performed a retrospective review of patients who underwent either one-shot IAC or NIAC before RC between October 2006 and November 2015. A propensity-score matching (1:3) was performed based on key characters. The Kaplan-Meier method was utilized to estimate survival probabilities, and the log-rank test was used to compare survival outcomes between different groups. A multivariable Cox proportional hazard model was used to estimate survival outcomes. RESULTS: Twenty-six patients were treated using IAC before RC, and 123 NIAC patients also underwent RC. After matching, there was no significant difference between groups in baseline characteristics, perioperative variables, complication outcomes or tumor characteristics. Compared with clinical tumor stages, pathological tumor stages demonstrated a significant decrease (P = 0.002) in the IAC group. There was no significant difference in overall survival (OS, p = 0.354) or cancer-specific survival (CSS, p = 0.439) between the groups. Among all patients, BMI significantly affected OS (p = 0.004), and positive lymph nodes (PLN) significantly affected both OS (p<0.001) and CSS (p = 0.010). CONCLUSIONS: One-shot neoadjuvant IAC before RC shows safety and tolerability and provides a significant advantage in pathological downstaging but not in OS or CSS. Further study of neoadjuvant combination therapeutic strategies with RC is needed.

Inhibition of autophagy enhances the anticancer effect of enzalutamide on bladder cancer.

BACKGROUND: Emerging preclinical evidence suggests a critical role for androgen-mediated androgen receptor (AR) signaling in bladder cancer progression. However, researchers have not determined whether autophagy modulates the efficacy of an enzalutamide (ENZ) treatment in subjects with advanced bladder cancer. In this study, we investigated the synergistic effect of ENZ and autophagy inhibitors on bladder cancer. METHODS: ENZ was used as an anti-AR drug, and chloroquine (CQ), 3-methyladenine (3-MA), and bafilomycin A1 (BAF) were used as autophagy inhibitors. J82, T24, and UMUC3 cell lines were used as models of bladder cancer. A bifluorescence autophagy system with the mRFP-GFP-LC3 plasmid was used to evaluate autophagy flux. Protein and mRNA levels were detected using Western blotting and qPCR, respectively. A Cell Counting Kit-8 (CCK-8) assay, colony assay, and flow cytometry analysis were used to evaluate cell proliferation and apoptosis. Four-week-old BALB/c athymic nude mice were used in the in vivo assay. RESULTS: Based on the results obtained using the bifluorescence autophagy system, ENZ (10-20 µM) significantly facilitated the accumulation of autophagosomes and autolysosomes in the cytoplasm of J82 and T24 cells. Additionally, ENZ significantly increased the expression of autophagy-related genes (AMP-dependent protein kinase (AMPK), autophagy-related gene 5 (ATG5), microtubule-associated protein light chain 3B (LC3B), and UNC-51-like kinase 1 (ULK1)) and proteins (microtubule-associated protein 1 light chain 3-II/I (LC3-II/I), ATG5, and phosphorylated AMP-dependent protein kinase α (p-AMPKα)). The administration of ENZ monotherapy (10-20 µM) to J82 and T24 cells failed to alter proliferation and apoptosis. Concurrent treatment with ENZ and autophagy inhibitors distinctly triggered apoptosis and inhibited proliferation. Genetic inhibition of autophagy by specifically blocking ATG5 with siRNA also increased ENZ-induced apoptosis in J82 and T24 cells. In vivo, concurrent treatment with ENZ (25 mg/kg/day) and CQ (10 mg/kg/day) improved the therapeutic sensitivity by decreasing tumor growth and apoptosis. Additionally, overexpression of AR suppressed ENZ-induced autophagy-related genes (LC3-II/I, ATG5, and p-AMPKα) in T24 cells, and CQ exerted synergistic effects with ENZ to suppressed AR-responsive genes expression (KLK2 and KLK3) in bladder cancer. In UMUC3 cells, ENZ monotherapy directly induced anticancer effects, and concurrent treatment with ENZ and CQ also had a synergistic effect on proliferation and apoptosis. CONCLUSIONS: Autophagy may be a potential mechanism underlying ENZ-resistant bladder cancer. Blockade of autophagy significantly increased ENZ-induced apoptosis in bladder cancer. Thus, concurrent treatment with autophagy inhibitors and ENZ may be a novel therapeutic strategy for bladder cancer.

Increased H2S and its synthases in urothelial cell carcinoma of the bladder, and enhanced cisplatin-induced apoptosis following H2S inhibition in EJ cells.

H2S, synthesized by cystathionine ß-synthase (CBS), cystathionine γ-lyase (CSE) and 3-mercaptopyruvate sulfurtransferase (MPST), functions as a signalling molecule in mammalian cells. H2S serves complex functions in physiological and pathological processes, including in bladder cancer. In the present study, H2S production, the expression of the associated enzymes and the effect of H2S on human urothelial cell carcinoma of the bladder (UCB) tissue and cell lines were evaluated, and whether decreasing H2S levels influenced cell viability and tumour growth following treatment with cisplatin (CDDP) was assessed in UCB cells in vitro and in vivo. H2S production and the expression of CBS, CSE and MPST in bladder tissue specimens and the UCB cell lines 5637, EJ and UM-UC-3 were analysed using a sulfur-sensitive electrode and western blotting. UCB cells were subjected to different treatments, and viability and protein expression were determined. H2S production was inhibited to examine its influence on EJ cell tumour growth following CDDP treatment in vivo. It was identified that CBS, CSE and MPST protein were up-regulated in UCB tissues and cells. The H2S production and enzyme expression levels were the highest in UCB tissue and EJ cells. The inhibition of endogenous H2S biosynthesis decreased EJ cell viability and tumour growth in response to CDDP treatment. H2S levels and the associated biosynthetic enzymes were increased in human UCB tissue and cells compared with adjacent tissue and normal cells, which may have increased the resistance to CDDP-induced apoptosis in UCB. Therefore, H2S and its production may be an alternative therapeutic target for UCB.

Expression of CD74 in bladder cancer and its suppression in association with cancer proliferation, invasion and angiogenesis in HT-1376 cells.

The aim of the present study was to investigate the expression and potential roles of CD74 in human urothelial cell carcinoma of the bladder (UCB) in vitro and in vivo. CD74 and macrophage migration inhibitory factor (MIF) were located and assayed in normal and UCB samples and cell lines using immunostaining. CD74 was knocked down using CD74 shRNA lentiviral particles in HT-1376 cells. The proliferative, invasive potential and microvessel density (MVD) of knockdown-CD74 HT-1376 cells were analyzed in vitro or in vivo. The expression of CD74 in an additional high grade UCB J82 cell line was also verified in vivo. All experiments were repeated at least 3 times. The majority of muscle-invasive bladder cancer (MIBC) samples, and only one high grade UCB cell line, HT-1376, expressed CD74, compared with normal, non-muscle-invasive bladder cancer (NMIBC) samples and other cell lines. The levels of proliferation and invasion were decreased in the CD74 knockdown-HT-1376 cells, and western blotting assay indicated that the levels of proteins associated with proliferation, apoptosis and invasion in the cells were affected correspondingly by different treatments in vitro. The tumorigenesis and MVD assays indicated less proliferation and angiogenesis in the knockdown-HT-1376 cells compared with the scramble cells. Notably, J82 cells exhibiting no signal of CD74 in vitro presented the expression of CD74 in vivo. The present study revealed the potential roles of CD74 in the proliferation, invasion and angiogenesis of MIBC, and that it may serve as a potential therapeutic target for UCB, but additional studies are required.

The clinical application of wrapping ureter by a pedicled gastrocolic omentum flap during cutaneous ureterostomy / 中华泌尿外科杂志

Objective To discuss the effect and value of wrapping ureter by a pedicled gastrocolic omentum flap during cutaneous ureterostomy as urinary diversion after radical cystectomy.Methods The clinical data of 12 cases(7 males and 5 females)with bladder cancer during January 2016 and September 2017 were retrospectively analyzed.The median age was 77 years (range 75-83),and the course of the disease was 5 days to 2 years.11 cases had gross hematuria,5 cases had frequent and urgent urination,1 case had lower abdominal pain,and 1 case had no obvious clinical symptoms.9 cases had a history of transurethral prostatic resection.8 cases had a complication of hypertension,5 cases had a complication of diabetes,7 cases had a complication of cardiovascular and cerebrovascular diseases,and 6 cases had a complication of anemia.The ASA level was 3 in 9 cases and was level 4 in 3 cases.All the patiants were diagnosed of bladder urothelial carcinoma by biopsy.There were 8 cases with T3 tumor and 4 cases with T4.All the patients underwent laparoscopic radical cystectomy under general anesthesia,and wrapping ureter by a pedicled gastrocolic omentum flap during cutaneous ureterostomy was used for urinary diversion.Take a segment of 3-5 cm wide pedicle omentum,which is covered from the distal end of the ureter to the end,and use a 4-0 absorbable line to fix the omentum and ureteral outer membrane,and then perform ureterocutaneostomy.The incidence of postoperative anastomotic stenosis,hydronephrosis and wound infection were observed.Results All the operations were performed successfully without obvious operative complications.The mean operating duration was125 (95-217) mins,estimated volume of blood loss 120 (50-600)ml.5 days after surgery,1 patient developed incision infection,which was cured by strengthening anti-infection drug therapy and wound dressing.No complications such as urinary fistula,intestinal fistula and peripheral hernia occurred.No death was reported during perioperative period.Single J tube was removed 1-3 months after surgery.Within a follow-up of 12 (3-23) months,1 case died of cerebral infarction 3 months after surgery,1 case occurred papilla light constriction without special treatment,no ureteral stenosis and necrosis cases and there were no cases of hydronephrosis.All the patients were satisfied with the quality of life after surgery.Conclusions Wrapping ureter by a pedicled gastrocolic omentum flap during cutaneous ureterostomy is simple,safe and effective as urinary diversion after radical cystectomy.

Comparison of perioperative outcomes and complications between intracorporeal and extracorporeal orthotopic ileal neobladder after laparoscopic radical cystectomy / 中华泌尿外科杂志

Objective To compare perioperative outcomes and complications between intracorporeal and extracorporeal orthotopic ileal neobladder after laparoscopic radical cystectomy (LRC) from one center.Methods We retrospectively analysis patients' clinical and follow-up data who underwent laparoscopic radical cystectomy (LRC) in our hospital from January 2011 to May 2017,of whom 25 cases underwent intracorporeal orthotopic ileal neobladder (ICIN) and 20 cases underwent extracorporeal orthotopic ileal neobladder (ECIN).The patients' characteristics,perioperative data,postoperative pathological results,postoperative complications and oncologic outcomes were compared and analyzed.The mean age of ECIN group was (59.9 ± 10.5)years,mean BMI was (24.7 ±3.3)kg/m2,the ASA level was less than Ⅱ in 19 cases and level Ⅲ in 1 case,the mean preoperative CCI was 4.5 ± 1.6,3 patients got preoperative neoadjuvant chemotherapy.The mean age of ICIN group was (59.0 ± 9.8) years,mean BMI was (25.3 ±4.3)kg/m2,the ASA level was less than Ⅱ in 25 cases and no level Ⅲ,mean preoperative CCI was 4.0 ±1.3,3 patients got preoperative neoadjuvant chemotherapy.There was no statistical difference between the two groups in patients' characteristics.All operations were performed under general anesthesia.After LRC was accomplished,an additional 12 mm trocar was placed 1 cm crania to the pubic symphysis for EndoGIA in ICIN group.An ileal segment 60 cm long was isolated 25 cm proximal to the ileocecum.The continuity of the small bowel was restored with side to side anastomosis.The proximal 10 cm was moved to the distal end of the ileal segment for the right isoperistaltic afferent limb,and the remaining proximal 10 cm ileal segment was reserved for the left isoperistaltic afferent limb.Then,the remaining length of the 40 cm ileal segment was detubularized along its antimesenteric border.The anterior wall of the neobladder was folded forward and the free edges were sutured to achieve a spherical configuration.After single J ureteric stents were inserted into the ureters,the urethra and ureters were sutured with neobladder successively.For ECIN,the ileal segment is removed via a midline laparotomy below the umbilicus 5-10 cm.The construction method of neobladder was the same with ICIN group.Results All 45 cases were successfully operated.There was no statistical difference between ICIN group and ECIN group in operation time [(374.2 ± 74.6) min vs.(360.3 ±83.6)min,P =0.557] and extraction time of pelvic drainage tube[(12.5 ±5.9)d vs.(11.8 ±4.5)d,P =0.686].Patients in the ICIN group had less intraoperative blood loss [(186.0 ± 140.3)ml vs.(272.0 ±110.7) ml,P =0.030],shorter oral intake time [(4.2 ± 2.2) d vs.(6.1 ± 1.6) d,P =0.002],shorter postoperative hospital stay [(13.7 ± 4.2) d vs.(19.9 + 7.6) d,P =0.001].There was no significant difference in the early postoperative complications (P =0.345).The median follow-up time of ICIN group and ECIN group were 30 months and 52.5 months,respectively.The day-time and night-time continence rate were 97.4％ and 86.8％ respectively in the first year after surgery without significant difference.A total of 6 patients suffered distant metastasis,of whom 4 patients suffered multiple metastases,1 case bone metastases and 1 case pelvic lymph node metastasis.Seven patients died,five of whom died of tumor recurrence or metastasis.There was no statistical difference between the two groups in cancer specific survival rates (P =0.644) and recurrence free survival rates (P =0.981).Conclusions Comparing with extracorporeal orthotopic ileal neobladder,intracorporeal orthotopic ileal neobladder has advantages of less blood loss and faster intestinal function recovery,without significantly increased operative time and complications.No significant difference was found in the function of neobladder and oncological outcomes.Randomized controlled trial with large sample and long-term follow-up is needed to verify the advantages of intracorporeal neobladder construction.

Laparoscopicradical cystectomy in the female:surgical tips and techniques / 中华泌尿外科杂志

Objective To discuss surgical tips and techniques of laparoscopic radical cystectomy in the female.Methods We retrospectively reviewed 29 females who underwent laparoscopic radical cystectomy between Feb.2008 and Mar.2017.The median age was 64 years (range 37-83 years),and the median BMI was 24.7 kg/m2 (range 12.1-31.2 kg/m2),and the median CCI was 3 (range 2-9).No distant metastasis was identified by bone scans,chest X-ray and sonography.Preoperative pathology were non-muscular infiltrating bladder cancer in 19 patients and muscular infiltrating bladder cancer in 10 patients by cystoscopic biopsy.Surgical method:under general anesthesia,the patients were placed in lithotomy position.A 5-port transperitoneal approach was used.Under laparoscopy,the suspensorium ligament of ovary was dissected and cut at lateral to the fimbriae of uterine tube,then the ureter was divided and umbilical artery was managed.The broad ligament of uterus was divided.Afterwards,the Douglas'pouch was opened and the fornix of vagina was cut transversely.Then,the endopelvic fascia was divided.The bladder was excised with the female reproductive organs together.If the female reproductive organs were decided to preserve,the dissection was performed along the space between bladder and uterus or vagina.Frozen section of urethra end andthe distal ureter were then obtained.The resected tissues were removed through the vagina,and then vagina was closed.Urinary diversion consisted of orthotopic ileal neobladder in 9 patients,ileal conduit in 14 patients,ureterostomy in 4 patients.All urinary tracts were removed for two cases due to renal function loss before surgery.Results Mean total operative time was 315 mins (ranged from180 mins to 420 mins).Estimated blood loss was 299 ml (ranged from 10 ml to 1 500 ml).Complications occurred in 2 cases.One patient had both vaginal infection and incomplete intestinal obstructionand the other had vaginal infection alone.No patients died in perioperative period.At a median follow up of 36.8 months (ranged from 2 to 110 months),24 patients (82.8％)were alive without recurrences,1 (3.4％) had metastasis in lymph nodes and 4 (13.8％) died.Conclusions Laparoscopic radical cystectomy in the female is technically feasible.Knowing female pelvic anatomy and grasping the techniques conduce to optimize the tips of the surgery and reduce complications.