VHL mutation drives human clear cell renal cell carcinoma progression through PI3K/AKT-dependent cholesteryl ester accumulation.

BACKGROUND: Cholesteryl ester (CE) accumulation in intracellular lipid droplets (LDs) is an essential signature of clear cell renal cell carcinoma (ccRCC), but its molecular mechanism and pathological significance remain elusive. METHODS: Enabled by the label-free Raman spectromicroscopy, which integrated stimulated Raman scattering microscopy with confocal Raman spectroscopy on the same platform, we quantitatively analyzed LD distribution and composition at the single cell level in intact ccRCC cell and tissue specimens in situ without any processing or exogenous labeling. Since we found that commonly used ccRCC cell lines actually did not show the CE-rich signature, primary cancer cells were isolated from human tissues to retain the lipid signature of ccRCC with CE level as high as the original tissue, which offers a preferable cell model for the study of cholesterol metabolism in ccRCC. Moreover, we established a patient-derived xenograft (PDX) mouse model that retained the CE-rich phenotype of human ccRCC. FINDINGS: Surprisingly, our results revealed that CE accumulation was induced by tumor suppressor VHL mutation, the most common mutation of ccRCC. Moreover, VHL mutation was found to promote CE accumulation by upregulating HIFα and subsequent PI3K/AKT/mTOR/SREBPs pathway. Inspiringly, inhibition of cholesterol esterification remarkably suppressed ccRCC aggressiveness in vitro and in vivo with negligible toxicity, through the reduced membrane cholesterol-mediated downregulations of integrin and MAPK signaling pathways. INTERPRETATION: Collectively, our study improves current understanding of the role of CE accumulation in ccRCC and opens up new opportunities for treatment. FUNDING: This work was supported by National Natural Science Foundation of China (No. U23B2046 and No. 62027824), National Key R&D Program of China (No. 2023YFC2415500), Fundamental Research Funds for the Central Universities (No. YWF-22-L-547), PKU-Baidu Fund (No. 2020BD033), Peking University First Hospital Scientific and Technological Achievement Transformation Incubation Guidance Fund (No. 2022CX02), and Beijing Municipal Health Commission (No. 2020-2Z-40713).

Parastomal Hernia Following Ileal Conduit: Incidence, Risk Factors, and Health-Related Quality of Life.

PURPOSE: The purpose of this study was to measure the incidence of parastomal hernia (PH) after radical cystectomy and ileal conduit. Secondary aims were the identification of risk factors for PH and to compare the health-related quality of life (QOL) between patients with and without PH. DESIGN: Retrospective review of medical records combined with cross-sectional administration of the QOL instrument and telephone follow-up. SUBJECTS AND SETTING: The study sample comprised 219 patients who underwent radical cystectomy and ileal conduit for urothelial cancer between February 2014 and December 2018. The study setting was Peking University First Hospital (Beijing, China). METHODS: Demographic and pertinent clinical data, including development of PH, were gathered via the retrospective review of medical records. Participants were also asked to complete the traditional Chinese language version of the City of Hope Quality of Life-Ostomy Questionnaire (C-COH). Multiple linear regression analysis was used to identify the effect of PH on C-COH scores. Logistic regression analysis was used to identify risk factors for PH development. RESULTS: At a median follow-up of 34 months (IQR = 21-48), 43 of 219 (19.63%) patients had developed a PH. A body mass index (BMI) indicating overweight (OR = 3.548; 95% CI, 1.562-8.061; P = .002), a prior history of hernia (OR = 5.147; 95% CI, 1.195-22.159; P = .028), and chronic high abdominal pressure postdischarge (CHAP-pd) (OR = 3.197; 95% CI, 1.445-7.075; P = .004) were predictors of PH after operation. There was no significant difference between C-COH scores of patients with or without PH. No significant differences were found when participants with PH were compared to those without PH on 4 factors of the C-COH: physical scores (ß= .347, P = .110), psychological scores (ß= .316, P = .070), spiritual scores (ß=-.125, P = .714), and social scores (ß= .054, P = .833). CONCLUSION: Parastomal hernia is prevalent in patients undergoing radical cystectomy and ileal conduit urinary diversion. Overweight, hernia history, and CHAP-pd were predictors of PH development. No significant differences in QOL were found when patients with PH were compared to those without PH.

The comprehensive complication index is more sensitive than the Clavien-Dindo classification for grading complications in elderly patients after radical cystectomy and pelvic lymph node dissection: Implementing the European Association of Urology guideline.

Objectives: Lack of assessment of 90-d perioperative morbidity in elderly patients after radical cystectomy and pelvic lymph node dissection (PLND) using a standard reporting methodology, and the Clavien-Dindo classification (CDC) does not accurately reflect the burden of complications. We aim to report the 90-d complications of elderly patients after radical cystectomy, and to compare the validity of the Comprehensive Complication Index (CCI) and CDC. Methods: Retrospective review of 280 patients aged ≥75 years who received radical cystectomy between 2006 and 2021. The 90-d complications of elderly patients after radical cystectomy were reported by implementing the EAU criteria. The CDC and CCI were both used for grading complications. The Spearman rank correlation coefficient was used to estimate the correlation between postoperative stay and CDC/CCI. Logistic regression was used to identify the risk factors for major complications. The sample size for a fictive superiority trial was calculated for different endpoints. Results: A total of 225 (80.36%) patients suffered from 528 complications. The cumulative CCI had a more accurate prediction of postoperative stay than the CDC (r = 0.378, p < 0.001 vs. r = 0.349, p < 0.001). The need for sample size could decrease when CCI was used for the primary endpoint. More risk factors for major complications were identified when CCI ≥33.7 was defined as the endpoint of major complications. Conclusion: CCI is better than CDC for grading the severity of complications in elderly patients after radical cystectomy and PLND.

Grb2 interacts with necrosome components and is involved in rasfonin-induced necroptosis.

The underlying mechanism by which growth factor receptor-bound protein 2 (Grb2) regulates necroptosis remains unexplored. In the present study, we found that rasfonin, a fungal natural product and an activator of necroptosis, enhanced Grb2 binding to receptor-interacting serine/threonine kinase 1 (RIP1), which plays a critical role in regulating programmed necrosis. Moreover, we observed that SQSTM/p62 (p62), a protein that can form necrosomes with RIP1, increased its interaction with Grb2 upon rasfonin challenge. Although it has been used as an activator of autophagy in our previous study, here we found that a high dose of rasfonin was able to inhibit autophagic process. Inhibition of RIP1 either chemically or genetically reversed the inhibition of rasfonin on autophagy, whereas knockdown of Grb2 markedly reduced rasfonin-induced necrosis. Additionally, we found that the compound failed to upregulate the expression of RIP1 in Grb2-deprived cells. In summary, our data revealed that Grb2 actively participated in rasfonin-induced necroptosis by interacting with the components of necrosome and mediating their expression.

Comparative Study of Perioperative and Oncological Outcomes Between Elderly Patients and Younger Patients Who Received Radical Cystectomy and Pelvic Lymph Node Dissection: A Single-Center Retrospective Study.

PURPOSE: To compare the perioperative and survival outcomes of patients over 75 years and younger patients who received radical cystectomy. PATIENTS AND METHODS: A total of 119 patients aged ≥75 years and 488 patients aged <75 years were enrolled. All patients underwent radical cystectomy with pelvic lymph node dissection. Clinical characteristics and perioperative outcomes were compared between the two groups. Overall survival and progression-free survival were analyzed by using the Kaplan-Meier method. Cox regression analysis and logistic regression analysis were used to identify the risk factors affecting the outcomes observed. RESULTS: There was no significant difference in perioperative complications between the elderly patient group and the younger patient group (p = 0.349). The 5-year overall survival of elderly patients was lower than that of young patients (p < 0.001). Age ≥75 years was a risk factor for overall survival (HR = 1.69 [95% CI: 1.22-2.35]; p = 0.002) and progression-free survival (HR = 1.69 [95% CI: 1.14-2.50]; p = 0.008) for patients who received radical cystectomy but was not a poor risk factor for major complications (HR = 1.25 [95% CI: 0.47-3.31]; p = 0.658) after radical cystectomy. In addition, preoperative renal insufficiency was associated with a higher risk of major complications. CONCLUSION: In our cohort, compared with younger patients, elderly patients aged ≥75 years had worse survival outcomes, but age ≥75 years was not a risk factor for major complications after radical cystectomy with pelvic lymph node dissection. Radical surgery should be encouraged for elderly patients who can tolerate aggressive treatments.

Laparoscopic versus open radical cystectomy in 607 patients with bladder cancer: Comparative survival analysis.

OBJECTIVES: To compare perioperative and oncologic survival outcomes between laparoscopic radical cystectomy and open radical cystectomy. METHODS: A total of 607 patients underwent open radical cystectomy (n = 412) or laparoscopic radical cystectomy (n = 195) at a single academic institution from January 2006 to April 2017. Their medical records were retrospectively analyzed. One-to-one propensity score matching was carried out to reduce selection bias. Estimated blood loss and complications were compared. Overall survival, cancer-specific survival and progression-free survival estimates for all patients and patients with locally advanced bladder cancer were analyzed using the Kaplan-Meier method. RESULTS: Either before or after matching, the laparoscopic radical cystectomy group had less estimated blood loss (P < 0.001 and P < 0.001) and fewer complications (P < 0.001 and P = 0.008). There was no difference in the overall survival (P = 0.216 and P = 0.961) and progression-free survival (P = 0.826 and P = 0.462) for all the patients having either laparoscopic radical cystectomy or open radical cystectomy. However, the 5-year progression-free survival of open radical cystectomy was higher than that of laparoscopic radical cystectomy (P = 0.019 and P = 0.021) for patients with locally advanced bladder cancer. CONCLUSIONS: Laparoscopic radical cystectomy is superior to open radical cystectomy in terms of perioperative outcomes, and similar to open radical cystectomy in terms of oncologic outcomes for patients with early stage bladder cancer. However, for patients with locally advanced bladder cancer, laparoscopic radical cystectomy seems to be associated with shorter progression-free survival than open radical cystectomy.

SQSTM1/p62 loss reverses the inhibitory effect of sunitinib on autophagy independent of AMPK signaling.

Sunitinib (ST), a multitargeted receptor tyrosine kinase inhibitor, has been demonstrated to be effective for the treatment of renal carcinoma. It has been reported that ST is involved in the mediation of autophagy; however, its regulatory role in the autophagic process remains controversial. Furthermore, the mechanism by which activated AMP-activated protein kinase (AMPK) negatively regulates autophagy remains nearly unexplored. In the present study, we revealed that ST inhibited AMPK activity and regulated autophagy in a cell type- and dose-dependent manner. In a number of cell lines, ST was demonstrated to inhibit H2O2-induced autophagy and the phosphorylation of acetyl-CoA carboxylase (ACC), whereas alone it could block the autophagic flux concurrent with increased expression of p62. An immunoprecipitation assay revealed that LC3 directly interacted with p62, whereas ST increased punctate LC3 staining, which was well colocalized with p62. Taken together, we reveal a previously unnoticed pathway for ST to regulate the autophagic process, and p62, although often utilized as a substrate in autophagy, plays a critical role in regulating the inhibition of ST in both basal and induced autophagy.

Is hysterectomy beneficial in radical cystectomy for female patient with urothelial carcinoma of bladder? A retrospective analysis of consecutive 112 cases from a single institution.

BACKGROUND: There is no criterion for determining whether female patients operated with cystectomy would benefit from hysterectomy. This study compares the oncological outcomes between female patients receiving uterus preserving cystectomy (UPC) and uterus excision cystectomy (UEC). METHODS: Retrospective review of 121 female patients with urothelial carcinoma of bladder undergoing UPC (n = 63) or UEC (n = 49) at a single institute between January 2006 and April 2017. Individual postoperative follow-up plans were performed for patients through outpatient visits. Overall survival (OS) and progression-free survival (PFS) estimates were analyzed using Kaplan-Meier method and multivariable Cox regression. RESULTS: The median follow-up time was 36 months (interquartile range 16-69). Among patients, 5 (4.1%) had uterus invasion. OS probability (p = 0.939) and PFS probability (p = 0.565) were similar in two groups. In multivariable Cox regression analysis, hysterectomy was not found to be a predictor of OS (hazard ratio 0.908, 95%CI 0.428-1.924, p = 0.801) and PFS (hazard ratio 1.109, 95%CI 0.439-2.805, p = 0.826) after adjusting for age, preoperative clinical stage, pathological stage, pathological nodal stage, neoadjuvant/adjuvant chemotherapy, location of the tumor, and surgical margin. No significant difference of overall survival probability was observed in the patients with organ-confined bladder cancer (p = 0.675) and in patients with no organ-confined bladder cancer (p = 0.695). CONCLUSIONS: The results showed that the rate of uterus invasion was low in patients analyzed in this cohort. It was also found that hysterectomy was not an independent predictor of OS and PFS after radical cystectomy in patients with bladder cancer.

Somatic mutations in renal cell carcinomas from Chinese patients revealed by whole exome sequencing.

BACKGROUND: While the somatic mutation profiles of renal cell carcinoma (RCC) have been revealed by several studies worldwide, the overwhelming majority of those were not derived from Chinese patients. The landscape of somatic alterations in RCC from Chinese patients still needs to be elucidated to determine whether discrepancies exist between Chinese patients and sufferers from other countries and regions. METHODS: We collected specimens from 26 Chinese patients with primary RCC, including 15 clear cell renal cell carcinoma (ccRCC) samples, 5 papillary renal cell carcinoma (PRCC) samples and 6 chromophobe renal cell carcinoma (ChRCC) samples. Genomic DNAs were isolated from paired tumor-normal tissues and subjected to whole exome sequencing (WES). Immunohistochemistry analysis was performed to detect the programmed death ligand 1 (PD-L1) expression in tumor tissues. RESULTS: A total of 1920 nonsynonymous somatic variants in exons and 86 mutations at splice junctions were revealed. The tumor mutation burden of ccRCC was significantly higher than that of ChRCC (P < 0.05). For both ccRCC and PRCC, the most frequent substitution in somatic missense mutations was T:A > A:T, which was different from that recorded in the COSMIC database. Among eight significantly mutated genes in ccRCC in the TCGA database, six genes were verified in our study including VHL (67%), BAP1 (13%), SETD2 (13%), PBRM1 (7%), PTEN (7%) and MTOR (7%). All the mutations detected in those genes had not been reported in ccRCC before, except for alterations in VHL and PBRM1. Regarding the frequently mutated genes in PRCC in our study, DEPDC4 (p.E293A, p.T279A), PNLIP (p.N401Y, p.F342L) and SARDH (p.H554Q, p.M1T) were newly detected gene mutations predicted to be deleterious. As the most recurrently mutated gene in ChRCC in the TCGA dataset, TP53 (p.R81Q) was somatically altered only in one ChRCC case in this study. The HIF-1 signaling pathway was the most affected pathway in ccRCC, while the PI3K-Akt signaling pathway was altered in all of the three RCC types. Membranous PD-L1 expression was positive in tumor cells from 6/26 (23%) RCC specimens. The PD-L1-positive rate was higher in RCC samples with the somatically mutated genes CSPG4, DNAH11, INADL and TMPRSS13 than in specimens without those (P < 0.05). CONCLUSIONS: Using WES, we identified somatic mutations in 26 Chinese patients with RCC, which enriched the racial diversity of the somatic mutation profiles of RCC subjects, and revealed a few discrepancies in molecular characterizations between our study and published datasets. We also identified numerous newly detected somatic mutations, which further supplements the somatic mutation landscape of RCC. Moreover, 4 somatically mutated genes, including CSPG4, DNAH11, INADL and TMPRSS13, might be promising predictive factors of PD-L1-positive expression in RCC tumor cells.

A novel adaptive ensemble classification framework for ADME prediction.

It has now become clear that in silico prediction of ADME (absorption, distribution, metabolism, and elimination) characteristics is an important component of the drug discovery process. Therefore, there has been considerable interest in the development of in silico modeling of ADME prediction in recent years. Despite the advances in this field, there remains challenges when facing the unbalanced and high dimensionality problems simultaneously. In this work, we introduce a novel adaptive ensemble classification framework named as AECF to deal with the above issues. AECF includes four components which are (1) data balancing, (2) generating individual models, (3) combining individual models, and (4) optimizing the ensemble. We considered five sampling methods, seven base modeling techniques, and ten ensemble rules to build a choice pool. The proper route of constructing predictive models was determined automatically according to the imbalance ratio (IR). With the adaptive characteristics of AECF, it can be used to work on the different kinds of ADME data, and the balanced data is a special case in AECF. We evaluated the performance of our approach using five extensive ADME datasets concerning Caco-2 cell permeability (CacoP), human intestinal absorption (HIA), oral bioavailability (OB), and P-glycoprotein (P-gp) binders (substrates/inhibitors, PS/PI). The performance of AECF was evaluated on two independent datasets, and the average AUC values were 0.8574-0.8602, 0.8968-0.9182, 0.7821-0.7981, 0.8139-0.8311, and 0.8874-0.8898 for CacoP, HIA, OB, PS and PI, respectively. Our results show that AECF can provide better performance and generality compared with individual models and two representative ensemble methods bagging and boosting. Furthermore, the degree of complementarity among the AECF ensemble members was investigated for the purpose of elucidating the potential advantages of our framework. We found that AECF can effectively select complementary members to construct predictive models by our auto-adaptive optimization approach, and the additional diversity in both sample and feature space mainly contribute to the complementarity of ensemble members.

Urodynamic characteristics of pelvic lipomatosis with glandular cystitis patients correlate with morphologic alterations of the urinary system and disease severity.

AIMS: To explore urodynamic characteristics and their clinical value in pelvic lipomatosis (PL) patients. METHODS: We reviewed the clinical information of 84 PL patients. A voiding pressure-flow study was used to classify patients into nonoutlet obstruction (NOO), latter-half-section obstruction (LHSO), or whole-section bladder outlet obstruction (BOO) groups. Urinary morphologic features were measured by imaging examination and cystoscopy. RESULTS: A unique LHSO that presented as sudden increasing detrusor pressure (Pdet) and decreasing flow rate in the latter half of voiding was observed for 52.4% (44 of 84) patients. Overall, 27.4% (23 of 84 patients) were diagnosed with BOO with whole-section increasing Pdet and decreasing flow rate. According to the morphologic feature analyses, the NOO patients had the largest angle of anteroposterior vesical walls (P < 0.001) and the least severe thickened bladder trigone (P = 0.015). The external compression at the bladder neck and thickened bladder trigone caused a prolonged and strictured bladder outlet tract (see the Supplementary video). There were 0, 5, and 4 urinary diversions performed in the NOO, LHSO, and BOO groups at diagnosis (P = 0.055). No patients in the NOO group, seven in the LHSO group, and two patients in the BOO group had disease progression at follow-up. Two LHSO patients and one BOO patients without hydronephrosis at diagnosis developed to hydronephrosis during follow-up. CONCLUSIONS: Morphologic alterations of the urinary system of PL patients lead to unique LHSO or BOO on UDS. The presences of LHSO and BOO are associated with disease severity and progression.

Comparison between completely and traditionally retroperitoneoscopic nephroureterectomy for upper tract urothelial cancer.

BACKGROUND: To evaluate the safety and efficacy of the completely retroperitoneoscopic nephroureterectomy (CRNU), a retrospectively comparative study between completely and traditionally retroperitoneoscopic nephroureterectomy (TRNU) was done in a single center. METHODS: From January 2014 to December 2014, 107 patients with upper tract urothelial cancer (UTUC) underwent CRNU. The kidney was retroperitoneoscopically dissected and the bladder cuff was cut by endoscopic gastrointestinal automatic stapler, and the specimen was removed from a 6-cm incision by posterior axillary line. Demographic, perioperative, and follow-up data were collected and compared retrospectively with 110 patients undergoing TRNU. RESULTS: The patients' characteristics between the two groups were not statistically different (p > 0.05), and all patients successfully received the procedure. The mean operative time (106 ± 37.9 versus 199 ± 69.1 min, p < 0.0001), the mean estimated blood loss (47.2 ± 82.4 versus 166.9 ± 250.9 ml, p = 0.002), and the mean hospital stay (6.1 ± 3.5 versus 8.1 ± 3.3 days, p = 0.03) of the CRNU group decreased significantly compared to the traditional group. The operative time was not affected by gender. No open conversion and major complications occurred. The surgical margin of the ureter was all negative. The mean follow-up time was 13.4 months for the CRNU group and 37.5 months for the TRNU group. All follow-up patients in the CRNU group were alive without local recurrence. No cases of port site metastasis and local recurrence were observed in both groups. Bladder tumor recurrence occurred in 4 patients of the CRNU group and 21 patients of the TRNU group. CONCLUSIONS: The CRNU using an endoscopic gastrointestinal automatic stapler to manage the bladder cuff is feasible and advantageous in decreasing the operative time, the blood loss, and the hospital stay. However, a larger sample and longer follow-up time will be still required.

Development of in Silico Models for Predicting P-Glycoprotein Inhibitors Based on a Two-Step Approach for Feature Selection and Its Application to Chinese Herbal Medicine Screening.

P-glycoprotein (P-gp) is regarded as an important factor in determining the ADMET (absorption, distribution, metabolism, elimination, and toxicity) characteristics of drugs and drug candidates. Successful prediction of P-gp inhibitors can thus lead to an improved understanding of the underlying mechanisms of both changes in the pharmacokinetics of drugs and drug-drug interactions. Therefore, there has been considerable interest in the development of in silico modeling of P-gp inhibitors in recent years. Considering that a large number of molecular descriptors are used to characterize diverse structural moleculars, efficient feature selection methods are required to extract the most informative predictors. In this work, we constructed an extensive available data set of 2428 molecules that includes 1518 P-gp inhibitors and 910 P-gp noninhibitors from multiple resources. Importantly, a two-step feature selection approach based on a genetic algorithm and a greedy forward-searching algorithm was employed to select the minimum set of the most informative descriptors that contribute to the prediction of P-gp inhibitors. To determine the best machine learning algorithm, 18 classifiers coupled with the feature selection method were compared. The top three best-performing models (flexible discriminant analysis, support vector machine, and random forest) and their ensemble model using respectively only 3, 9, 7, and 14 descriptors achieve an overall accuracy of 83.2%-86.7% for the training set containing 1040 compounds, an overall accuracy of 82.3%-85.5% for the test set containing 1039 compounds, and a prediction accuracy of 77.4%-79.9% for the external validation set containing 349 compounds. The models were further extensively validated by DrugBank database (1890 compounds). The proposed models are competitive with and in some cases better than other published models in terms of prediction accuracy and minimum number of descriptors. Applicability domain then was addressed by developing an ensemble classification model to obtain more reliable predictions. Finally, we employed these models as a virtual screening tool for identifying potential P-gp inhibitors in Traditional Chinese Medicine Systems Pharmacology (TCMSP) database containing a total of 13 051 unique compounds from 498 herbs, resulting in 875 potential P-gp inhibitors and 15 inhibitor-rich herbs. These predictions were partly supported by a literature search and are valuable not only to develop novel P-gp inhibitors from TCM in the early stages of drug development, but also to optimize the use of herbal remedies.

Inhibition of androgen induces autophagy in benign prostate epithelial cells.

OBJECTIVE: 5-α Reductase inhibitor can reduce the volume of benign prostatic hyperplasia by lowering benign prostatic hyperplasia level and consequently inducing epithelial cells apoptosis. The present study investigated whether autophagy and apoptosis of benign prostatic hyperplasia epithelial cells are influenced by low benign prostatic hyperplasia levels. METHODS: PWR-1E prostate epithelial cells transfected with GFP-LC3 plasmid were subjected to androgen deprivation conditions. Then the autophagic puncta were evaluated by fluorescence microscopy, and the cellular apoptosis rate was detected by 4, 6-diamidino-2-phenylindole staining after blocking of autophagic process by 3-methyladenine. Furthermore, autophagy status was also determined in hyperplasia prostate tissues from 5-α reductase inhibitor-treated patients by immunohistochemistry. RESULTS: In the androgen deprivation medium, autophagic punta increased markedly in PWR-1E cells, and blockage of autophagy by 3-methyladenine significantly promoted PWR-1E cells' apoptosis rate. In vivo, the expression of LC3 protein (an important autophagic marker) in hyperplasia prostate tissue significantly increased after 5-α reductase inhibitor treatment. Meanwhile, the prostate-specific antigen, as an inner control, decreased. CONCLUSION: 5-α Reductase inhibitor treatment increases autophagy and possibly decreases the apoptosis of prostate epithelial cells.

[Cisplatin enhances apoptosis in bladder cancer cells via autophagy].

OBJECTIVE: To monitor the cisplatin-induced autophagy and investigate the function of autophagy in bladder cancer cells. METHODS: The transmission electron microscope was used to detect autophagic vacuoles and the fluorescence microscope to detect GFP-LC3. The expressions of proteins, such as LC3, PARP, mTOR, P70S6K were analyzed by immunoblotting. Cell viability was analyzed by MTS assay, in which rapamycin was used to inhibit mTOR phosphorylation and enhance autophagy. LC3 expression was knocked down by RNA interference. RESULTS: In bladder cancer cell T24, autophagic vacuoles were observed by the transmission electron microscope and GFP-LC3 aggregation was viewed by the fluorescence microscope after cisplatin treatment. The LC3-II accumulation was enhanced by cispaltin treatment. Particularly at the concentrations of 50, and 100 µmol/L for 48 h , the gray value of LC3-II/Actin(%) increased 30 and 44, respectively. Cisplatin treatment inhibited the phosphorylation of mTOR/P70S6K, which was most significant at the concentration of 100 µmol/L for 48 h. Cisplatin also induced cell viability loss, which was 12% and 45% at the concentrations of 50, and 100 µmol/L for 24 h. This effect could be enhanced by rapamycin (F=74.890,P<0.01). Furthermore, knocking down LC3 by RNA interference reduced PARP cleavage. CONCLUSION: Cisplatin could induce autophagy in bladder cancer cell T24, which promoted cisplatin-induced apoptosis.

Measuring diagnostic accuracy of imaging parameters in pelvic lipomatosis.

OBJECTIVES: To study whether the individual radiological findings can help predict diagnosis of pelvic lipomatosis (PL) or, specifically appreciate its progression. METHODS: Data from 32 clinically proven cases of PL and 25 controls were collected. Two reviewers were recruited for a blinded evaluation, image features were recorded in terms of: (1) bladder shape; (2) bladder-rectosigmoid morphological indexes including ratio of superior-inferior to anterior-posterior length of bladder (SI/AP), angle between anterior and posterior wall (AAP), relative length of posterior urethra (rLPU), angle between bladder and seminal vesicle (ABS) and rectosigmoid morphological index (RMI); (3) secondary complications. Results were evaluated by an unpaired t test and ROC analysis. RESULTS: The sensitivity and specificity were 40.6% and 100% for pear and banana-shaped bladder, 62.5% and 100% for SI/AP, 40.6% and 100% for AAP, 62.5% and 100% for ABS, 78.1% and 72% for rLPU, 59.4% and 96% for RMI, respectively. These radiological findings partially correlated with the severity of disease weighted by hydronephrosis and treatment grade. Image analysis demonstrated high prevalence of glandular cystitis (100%) and hydronephrosis (73.4%). CONCLUSION: We conclude that PL is a progressive disease involving multiple pelvic organs with high prevalence of intractable cystitis and hydronephrosis. The imaging characteristics can help predict diagnosis and, specifically appreciate progression.

[Risk factor analysis of post-operative complications in radical cystectomy for bladder transitional cell cancer: retrospective assessment from 208 consecutive cases].

OBJECTIVE: To describe early complications (within 90 days) after radical cystectomy and to analyze the associated specific risk factors. METHODS: The clinical data from 208 consecutive cases of muscle invasive bladder cancer were collected and reviewed. Potential variables predictive of early morbidity were retrospectively analyzed. RESULTS: Of the 208 subjects, 160 (76.9%)developed at least 1 postoperative complication and 46 (22.2%) at least 2 complications. The most frequent complications presented were blood loss (75 cases), post-operative renal insufficiency (31 cases), and intestinal obstruction (29 cases). In univariate analysis, operative time, hypertension and preoperative creatinine level were associated with the development of complications. On multivariate analysis, operative time, preoperative creatinine level were the significant factors. CONCLUSION: Morbidity remains high after radical cystectomy. The operative time, preoperative creatinine level and hypertension may be associated with the postoperative complications. Acknowledgement of the patients' specific risk factors and monitoring perioperative processes may incrementally reduce risks and improve outcomes of the patients.

[Clinical significance of radical cystectomy with extended lymphadenectomy].

OBJECTIVE: To evaluate the prognosis of patients treated with radical cystectomy and extended lymphadenectomy for invasive bladder cancer and to describe the association of the primary bladder tumor grade, stage, lymph node status and lymph node density with clinical outcomes. METHODS: A retrospective analysis was done of 32 consecutive cases with bladder cancer who received radical cystectomy and extended lymph node dissection from January 2006 to December 2010 in the Department of Urology, Peking University First Hospital. All the patients were scheduled for the follow-up. The survival data were analyzed with the tumor grade, stage, lymph node status and other factors that might relate to the prognosis by statistics. RESULTS: All the cases were diagnosed as invasive urothelial bladder cancer by preoperative biopsy or TUR-Bt. During the follow-up, 6 patients (18.8%) got disease progression, and 4 patients died (12.5%). Overall survival rate was 87.5%. In 32 patients, 17 months and 3 year survival rates were (88.7 ± 12.1)% and (81.8 ± 17.0)%, respectively. From the tumor grade point of view, 6 patients belonged to G2, and 26 cases G3. All deaths were graded G3. G3-class 3-year survival rate was (74.6 ± 23.9)% (P> 0.05, compared with G2). From the analysis of stage, T1 and T2 stages had no death during the follow-up. The 17-month survival rate of T3 group was (60.0 ± 42.9)%, the 8-month survival rate of T4 group was (87.5 ± 22.9)%. There were 9 patients with positive lymph nodes (28.1%) and 23 with negative in (71.9%). The number of dissection lymph nodes was from 7-50 ( average 22.8 ). The 17-month survival rate of patients with positive lymph node was (50.0 ± 44.6)%. The 3-year survival rate of patients with negative lymph node was (92.3 ± 14.5)% (P<0.05). The 3-year survival rate of patients with lymph node density less than 20% was (88.8 ± 15.4)%. The 8-month survival rate of patients with lymph node density greater than 20% was (66.7 ± 53.3)% (P<0.05). CONCLUSION: Radical cystectomy with extended lymph node dissection can improve the prognosis of patients with invasive bladder cancer. Tumor stage, lymph node status and lymph node density are closely related to the patient's survival.

[Viviperception on renal vessel variation by retroperitoneal laparoscopy a report of 525 cases].

OBJECTIVE: To study the variation of renal vessels with retroperitoneal laparoscopy so as to increase the safety of retroperitoneal laparoscopic surgeries. METHODS: A total of 525 patients underwent retroperitoneal laparoscopic nephrectomy or partial nephrectomy at our hospital between January 2004 and June 2008. There were 316 males and 209 females with a mean age of (58 ± 13) years old. The procedures were as follows: (1) patients lay on one side with their waist up and the retroperitoneal cavity was established with our institutional method; (2) gerota's fascia was separated widely along the ventral surface of major psoas muscle; (3) the tissues around renal arteries and veins were isolated by ultrasonic scalpel. Careful observation was performed to explore if there were duplicated or accessory renal vessels; (4) renal vessels were cut by Endo-GIA/Hem-o-lok or blocked by bulldog clamps; (5) whole or partial kidney was finally resected (remaining procedures omitted). RESULTS: Among all patients, 58 patients (11.0%, 58/525) had a variation of renal vessels. There were double renal arteries on one side (n = 18), double renal veins (n = 10), 3 renal veins (n = 1) and double arteries and veins on one side (n = 3). Twenty-five patients (4.8%, 25/525) had one accessory renal artery on one side while 19 (76.0%, 19/25) accessory renal arteries went toward the upper kidney pole. The diameter of one patient's left spermatic vein was similar with that of renal vein and they were joined by lumber vein. CONCLUSION: The variation of renal artery is more common than that of renal vein. The accessory renal arteries are common and usually go toward the upper kidney pole. The variation of renal vessels should be considered before and during a laparoscopic procedure.