MTHFR C677T polymorphisms are associated with aberrant methylation of the IGF-2 gene in transitional cell carcinoma of the bladder

The purpose of this study was to determine the relationship between methylation status of the insulin-like growth factor 2 （IGF-2） gene and methylenetetrahydrofolate reductase （MTHFR） C677T gene polymorphisms in bladder transitional cell carcinoma tissues in a Chinese population. The polymorphisms of the folate metabolism enzyme gene MTHFR were studied by restrictive fragment length polymorphism （RFLP）, PCR-based methods of DNA methylation analysis were used to detect the CpG island methylation status of the IGF-2 gene. The association between the methylation status of the IGF-2 gene and clinical characteristics, as well as MTHFR C677T polymorphisms, was analyzed. Aberrant hypomethylation of the IGF-2 gene was found in 68.3% bladder cancer tissues and 12.4% normal bladder tissues, respectively, while hypomethylation was not detected in almost all normal bladder tissues. The hypomethylation rate of the IGF-2 gene in cancer tissues was significantly higher in patients with lymph node metastasis than in those without lymph node metastasis （46.3% vs 17.2%, P = 0.018）. No association was found between aberrant DNA methylation and selected factors including sex, age, tobacco smoking, alcohol consumption and green tea consumption. After adjusting for potential confounding variables the variant allele of MTHFR C677T was found to be associated with hypomethylation of the IGF-2 gene. Compared with wildtype CC, the odds ratio was 4.33 （95% CI=1.06-10.59） for CT and 4,95 （95% CI=1.18-12.74） for TT. MTHFR 677 CC and CT genotypes might be one of the reasons that cause abnormal hypomethylation of the IGF-2 gene, and the aberrant CpG island hypomethylation of the IGF-2 gene may contribute to the genesis and progression of bladder transitional cell carcinoma.

RECURRENCE RISK FACTORS IN PATIENTS WITH TRANSITIONAL CELL CARCINOMA OF BLADDER

Objective： To study recurrence factors and set up a model to evaluate the prognosis of patients with bladder cancer. Methods： An analysis on recurrence-related factors was made by Cox＇s proportional hazards model analysis and logistic multiple linear regression model analysis in 212 patients with transitional cell carcinoma treated surgically from 1995-2001. These factors included clinical and pathologic figures. Results： The most important factor is metastasis to the regional lymph nodes, the Hazards ratio is 6.6 （P=0.0004）, followed by multiple tumors （Hr=2.255, P〈0.0001）, tumor in trigone and bladder neck （Hr=2.053, P〈0.0001）, stage （Hr=2.057, P〈0.0001）, grade （Hr=1.569, P=0.0081）, intravesical chemotherapeutic instillations （Hr-0.559, P=0.0011） and hematuria （Hr=0.762, P=0.0076）. A predicting equation was established, and the predicting values were calculated according to the individual features of patients. The predicting and actual values were compared, and the sensitivity, specificity and overall concordance were 83.5%, 67.6% and 80.1% respectively. Conelusion：The evaluation of prognosis could be made quite accurately based on these factors.

Cloning of Human Uroplakin Ⅱ Gene from Chinese Transitional Cell Carcinoma of Bladder and Construction of Its Eukaryotic Expression Vector

Summary: To clone Uroplakin Ⅱ gene from Chinese transitional cell carcinoma (TCC) of bladder and construct its eukaryotic expression vector, the molecular cloning method was used to extract total RNA from a GⅢ/ T 3N 0M 0 tissue sample of the bladder TCC patients. The primers were designed by Primer 5.0 software. Full length cDNA of Uroplakin Ⅱ gene was amplified by reverse transcription polymerase chain reaction (RT-PCR), assayed by nucleic acid sequencing and then inserted between XbaⅠ and HindⅢ restrictive sites of eukaryotic expression vector pcDNA3.0. The recombinant was assayed by restricted enzyme digestion. Under the induction of Lipofectamine 2000, the recombinant was transfected into Uroplakin Ⅱ negative bladder cancer cell line EJ. Cellular expression levels of Uroplakin Ⅱ were detected by RT-PCR. The nucleic acid sequencing results indicated that Chinese Uroplakin Ⅱ cDNA (555 bp) was successfully cloned. The BLAST analysis demonstrated that the cloned sequence is 100 % homologous with sequences reported overseas. The GenBank accession number AY455312 was also registered. The results of restricted enzyme digestion indicated that eukaryotic vector pcDNA-UPⅡ for Uroplakin Ⅱ was successfully constructed. After being transferred with pcDNA-UPⅡ for 72 h, cellular Uroplakin Ⅱ mRNA levels were significantly improved (P<0.01). It is concluded that human Uroplakin Ⅱ gene was successfully cloned from Chinese TCC tissues, which provided a basis for further exploration of the roles of Uroplakin Ⅱ gene in TCC biological behaviors and potential strategies for targeted biological therapy of TCC.

Research on the prognosis of different types of microvessels in bladder transitional cell carcinoma

BACKGROUND At present,there is controversy on the role of microvessel density(MVD)in tumors as a prognostic indicator of bladder transitional cell carcinoma(BTCC).However,the MVD in tumors is simply classified based on the expression of several different vascular markers,which has not been related to analytical research on the prognosis of patients with BTCC.AIM To explore the classification of blood vessels in tumors and studied the relationship between MVD and the prognosis of patients with BTCC.METHODS The tissue mass was detected by tissue microarray and immunohistochemical analysis with monoclonal antibodies against CD31,CD34,CD105,and vascular smooth muscle actin to investigate the MVD in BTCC.The measurement data are expressed as the mean±SD.The difference between the groups was analyzed by the t-test,the counting data were analyzed byχ2 test.The Kaplan-Meier survival curve was estimated by the product-limit method.The log-rank time-series test was employed to compare the tumor-free survival curves.RESULTS The MVD was closely related to the pathological grade,invasive depth,and prognosis of BTCC.Significant differences were found between grade I and grade II,grade II and grade III,superficial and invasive type,and the tumor-free survival group and the recurrence or metastasis group(P<0.01).Multivariate analysis showed that undifferentiated MVD was an independent prognostic factor for patient survival time.An inverse correlation between undifferentiated tumor MVD and differentiated tumor MVD in BTCC was also shown.CONCLUSION The classification of blood vessels in BTCC could act as an important prognostic indicator and may also be of great significance in the treatment of cancer.

Long-term follow-up of Ta transitional cell carcinoma of bladder after treatment of TURBt plus intravesical therapy

To stduy the association between the prognosis of Ta transitional cell carcinoma (TCC) of the bladder and risk-related factors.Methods A total of 88 cases (62 males and 26 females;mean age,61 years;age range,41-81 years) of initial T.TCC of the bladder treated with transurethral resection of bladder tumor (TURBt) plus intravesical chemotherapy or immunotherapy were enrolled.Among them,there were 26 cases of G1,61 cases of G2 and 1 case of G3.For tumor site,62 cases (16 cases of G1,45 of G2,1 of G3) had single tumor and 26 cases (10 cases of G1,16 of G2) had multi-site tumors.The mean follow-up was 113 months (range,56-168 months).The tumor grade,original tumor number and their association with the recurrence and progression of this type of TCC were retrospectively analyzed.Results The overall recurrence rate (RR) was 60% (53/88).In single tumor group,RR of G1 cases was 25% (4/16);RR of G2 cases was 62% (28/45) and the total RR was 52% (32/62).In multi-site tumor group,RR of G1 cases was 80% (8/10),RR of G2 cases was 75% (12/16) and the total RR was 77% (20/26).The RR of multi-site tumor group was significantly higher than that of single tumor group (P<0.01).In single tumor group,RR of G2 cases was significantly higher than that of G1 cases (P<0.001).In multi-site tumor group,there was no association of RR with tumor grade.There was no progression in G1 tumor cases.The progression rate was 42.5% (17/40) in G2 tumor cases;among them,30% (12/40) progressed to T1G2 tumors and 12.5% (5/40) progressed to T2G2 tumors.The RR of cases who received thiotepa,mitomycin and BCG were 75% (12/16),68% (30/44) and 40% (11/27),respectively.Tumor specific mortality was 1.14% (1/88,a T2G3 case).Conclusion The multi-site Ta TCC of the bladder has relatively higher RR and greater chance of progression after the treatment of TURBt plus intravesical chemotherapy or immunotherapy,especially in the poor differentiated tumors,thus active treatment and close follow-up are essential in clinical practice.9 refs.

Survivin mRNA expression in urine as a biomarker for patients with transitional cell carcinoma of bladder

Transitional cell carcinoma （TCC） of bladder is the most common malignant tumor in uropoiesis system. Up to date, there is still lack of an ideal marker for the diagnosis of TCC except CT and MRI imaging and cystoscopy. Cystoscopy is an invasive examination, which increases the possibility of urinary tract infection. Urine cytology has low sensitivity （21%--40%） in diagnosis of bladder cancer, especially for those with medium or high differentiation. The specificity is often affected by factors such as specimen collection, urinary tract infection, etc. Detecting the expression of survivin mRNA in urine by real time-PCR is simple in specimen collection and is sensitive and relatively specific, which provides a simple and noninvasive diagnostic method for TCC. Moreover it allows comparing the gene expression levels at different stages and grades of TCC, which can help define malignancy degree of TCC.

Transitional cell carcinoma associated with aristolochic acid nephropathy： most common cancer in chronic hemodialysis patients in China

Background The research of cancer in patients on hemodialysis （HD） in China has not been reported. The aim of this study was to investigate the clinical and histological features and outcomes of cancer in Chinese HD patients. Methods The study subjects were 49 cancer patients （1.4%） out of 3448 end stage renal disease （ESRD） patients maintained on HD at China-Japan Friendship Hospital from October 1997 to July 2011. Results Urinary tract cancer （74%） was the most common followed by gastrointestinal tract cancer （12%）, breast cancer （6%）, lung cancer （4%）, thyroid cancer （2%）, and hematologic cancer （2%）. Thirty-three patients （67%） had urinary tract transitional cell carcinoma （TCC） and 29 of them had aristolochic acid nephropathy （AAN） as underlying disease. Death occurred in eight patients out of 49, and the survival rate of HD patients with cancer was similar to those without cancer （P=0.120）. Conclusion The urinary tract TCC is the most common cancer in HD patients with AAN in one of the centers of northern China.

Prognostic factors for primary superficial transitional cell carcinoma of the bladder:a retrospective cohort study

Background Previous studies showed that the prognostic factors for superficial transitional cell carcinoma of the bladder varied with the findings of different cohorts. Few multivariate analyses of prognostic factors for superficial bladder tumors have been reported in China and bladder preservation as a prognostic index of superficial bladder tumors is limited and scarce in Chinese patients. This study was conducted to analyze a group of risk factors for prognostic outcomes for patients with primary superficial transitional cell carcinoma of the bladder. Methods Between January 1980 to December 2000, 198 patients [172 men and 26 women; mean age （52.98± 11.28） years] with primary superficial transitional cell carcinoma who were pathologically classified as Ta or T1 in Hunan Provincial Tumor Hospital （Changsha, China） were enrolled in this study. Surgical methods included local resection and electric coagulation of bladder tumors, transurethral resection of bladder tumors and partial cystectomy. After initial surgical treatment, patients were followed through a cystoscopy every three months during the first two years and every six months thereafter in the design of retrospective cohort. Survival analysis was performed to analyze risk factors of the prognostic outcomes for transitional cell carcinoma of the bladder. Canonical correlation analysis was conducted to present and interpret synthetically the multi-correlation between all kinds of prognostic outcomes and risk factor in multiply dimensions. Results The average follow-up period was （6.65±4.74） years. Assessments at three, five, and 10 years showed recurrence rates, respectively, of （28.32±3.45）%, （35.31±3.83）%, and （42.48±4.40）%; progression rates of （8.89±2.14）%, （15.16±2.94）%, and （23.88±4.19）%; bladder-preservation rates of （94.68± 1.74）%, （93.87± 1.91）%, and （91.51±2.49）%; metastasis rates of （8.25＋2.05）%, （11.24±2.47）%, and （28.94±4.93）%; and cancer-related survival rates of （95.02± 1.62）%, （90.70±2.45）%, and （77.14±4.88）%. The main risk factors for recurrence were histological grade, blood transfusion during surgery and the duration of symptoms. Progression was affected by blood transfusion during surgery, histological grade, the number of re-examinations, and the length of the recurrence-free period. Metastasis was associated with tumor multifocality, hydronephrosis, microscopic growth pattern, and the recurrence-free period. Cancer-related survival was influenced by microscopic growth pattern and the recurrence-free period. Bladder preservation involved only the recurrence-free period. The comprehensive results from canonical correlation analysis showed that the main prognostic outcomes were cancer-related survival, metastasis and progression respectively, while the dominate risk factors were histological grade, tumor multifocality and blood transfusion. Conclusions The risk factors were different for each prognostic outcome of transitional cell carcinoma of the bladder. This is helpful for predicting the prognosis of transitional cell carcinoma of the bladder and designing therapeutic and follow-up strategies for this cancer.

Poor visualization of renal collecting system in intravenous urography as an indicator of invasive transitional cell carcinoma in the upper urinary tract

Background Transitional cell carcinoma of the upper urinary tract （UUT-TCC） accounts for 5% to 10% of all renal tumours and 5% to 6% of all urothelial tumours all over the world. In China, the proportion of UUT-TCC to all urothelial tumours may be 26%, which is higher than that in the western world. The early diagnosis of UUT-TCC is difficult and the present study elucidates the diagnostic value of poor or nonvisualization （PNV） in intravenous urography in patients with UUT-TCC and its correlations with pathological findings and clinical characteristics. Methods The data of 172 consecutive patients between January 1997 and January 2005 with UUT-TCC who underwent nephroureterectomy in our departments were selected and analyzed retrospectively. Results Of our sample, 144 cases presented with gross haematuria （83.7%） and 12 with microscopic haematuria （7.0%）. Forty-six cases （26.7%） were detectable by cytology. Filling defect identified 36 positive cases of 172 patients （20.9%）, PNV was present in the images of 105 of 172 patients （61.0%）. The detection rate by PNV （61.0%） was significantly different from that by cytology （26.7%） or by filling defect （20.9%） （P=-0.031, P=-0.001, respectively）. Univariate logistic regression analysis for PNV showed that tumour stage, grade and size were significant predictors （P=0.028; P=-0.031; P=-0.006, respectively）. Tumour stage and size were identified as independent risk factors in the multivariate logistic regression model （P=-0.042; P=-0.014）.Conclusions Except for suspected urolithiasis, urinary tuberculosis or congenital abnormalities, UUT-TCC should be considered if PNV exists in intravenous urography especially of old patients. The value of PNV is much more significant than filling defect in intravenous urography in the diagnosis of UUT-TCC. It is supposed that PNV carries more risk of higher stage and larger tumour size in UTT-TCC.

Heat shock protein 70 expression in relation to apoptosis in primary bladder transitional cell carcinoma

Bladder carcinoma is the most common tumor in the urinary system. In 1996, a sampleinvestigation showed that bladder carcinoma, in which more than 90% was mainly primary bladder transitional cell carcinoma （BTCC）, was one of the ten highest mortality malignant tumors in China. Bladder carcinoma represented 2% of all malignant tumors and has the fifth most common malignancy in men in Europe and North America.

Clinical implications of single cell sequencing for bladder cancer

Bladder cancer(BC)is the 10th most common cancer worldwide,with about 0.5 million reported new cases and about 0.2 million deaths per year.In this scoping review,we summarize the current evidence regarding the clinical implications of single-cell sequencing for bladder cancer based on PRISMA guidelines.We searched PubMed,CENTRAL,Embase,and supplemented with manual searches through the Scopus,and Web of Science for published studies until February 2023.We included original studies that used at least one single-cell technology to study bladder cancer.Forty-one publications were included in the review.Twenty-nine studies showed that this technology can identify cell subtypes in the tumor microenvironment that may predict prognosis or response to immune checkpoint inhibition therapy.Two studies were able to diagnose BC by identifying neoplastic cells through single-cell sequencing urine samples.The remaining studies were mainly a preclinical exploration of tumor microenvironment at single cell level.Single-cell sequencing technology can discriminate heterogeneity in bladder tumor cells and determine the key molecular properties that can lead to the discovery of novel perspectives on cancer management.This nascent tool can advance the early diagnosis,prognosis judgment,and targeted therapy of bladder cancer.

ESTABLISHMENT AND CHARACTERIZATION OF TWO CELL LINES DERIVED FROM HUMAN TRANSITIONAL CELL CARCINOMA

Two continuous cell lines derived from the neoplastic urothelium had been maintained in culture for more than two years. The first cell line derived from the urothelium of a fusion papillocarcinoma on the left lateral wall of the bladder was designated as TBC-1 and grown in vitro for more than 150 generations. The second cell line derived from the urothelium of a papillocarcinoma in the left renal pelvis was designated as TPC-1 and grown in vitro for more than 100 generations. Characterization studies made on both cell lines showed that the cells had a rapid doubling time, exhibited mul-tilayering and produced tumors in sc of BALB / c. Tumor nodules that produced sc of BALB / c kept similar cellular and pathological features to those of the primary biopsy specimens under light and electron microscopes. TPC-1 cell line exhibited a three-dimensional structure of transitional epithelium on the nylon-mesh disk which was coated with a layer of rat tail collagen. Both TBC-1 and TPC-1 cell lines formed colonies in soft agar. Their forming rates were 35% and 28%, respectively. The chromosome number of TBC-1 cells ranged from 17 to 84, with a modal number of 54; whereas that of TPC-1 cells ranged from 28 to 139, with a modal number of 49. The TBC-1 cells showed mutant p53 and ras p21 protein expression and expressed weakly ABH blood group isoantigens. Analysis of lactic dehydrogenase (LDH) isozymes showed the highest levels of LDH isozyme 4 sonicated cell lysates of TBC-1 and TPC-1 respectively.

Expression and clinical implication of PRL-1 and PRL-3 in transitional cell carcinoma of bladder

The mRNA and protein expression of phos-phatase of regenerating liver 1(PRL-1)and phosphatase of regenerating liver 3(PRL-3)in transitional cell carcinoma of bladder(BTCC)and normal epithelia of bladder was investigated,and the relationship between the BTCC and pathological changes was clarified.The expression of PRL-1 and PRL-3 mRNA was detected by using reverse transcription polymerase chain reaction(RT-PCR)in 30 cases of BTCC and 10 cases of normal bladder,and the expression of PRL-1 and PRL-3 protein was checked by using immunohistochemistry in 30 cases of BTCC and 15 cases of normal bladder.The expression levels of PRL-1 and PRL-3 mRNA and protein were higher in BTCC than those in normal bladder epithelia(P<0.05).The increased expression of PRL-1 and PRL-3 mRNA and protein was detectable in deep invasion and metastasis of BTCC(P<0.05).There was no correlation between the expres-sion of PRL-1 and PRL-3 and gender,age or recurrence of BTCC(all P>0.05).A significantly positive correlation was found between PRL-1 and PRL-3 in BTCC(P<0.05).PRL-1 and PRL-3 are expressed consistently and may contribute to the growth,differentiation,invasion and metastasis of BTCC.

Simultaneous robot-assisted approach in a super-elderly patient with urothelial carcinoma and synchronous contralateral renal cell carcinoma:A case report

BACKGROUND It is rare for urothelial and renal cell carcinomas to coexist in the same patient,and even rarer for them to be detected simultaneously.Because of this rarity,a standard treatment has not been established and studies about overall survival are scarce.Therefore,physicians must modify treatments according to the individual’s situation and the stage of each disease.In recent years,with advances in the instruments and techniques,minimal invasive robotic surgeries have become available for advanced-stage or high-risk patients.CASE SUMMARY An 85-year-old woman with a medical history of hypertension and hyperlipidemia visited our institution.She had visited her local hospital complaining of intermittent,painless,gross hematuria that had started 3 mo earlier.On computed tomography,a right renal mass and left proximal ureteral mass with hydronephrosis were found simultaneously.We decided to perform robot-assisted surgery on both sides during one operation.Considering renal function and kidney loading,right partial nephrectomy was performed first,followed by left nephroureterectomy with bladder cuff excision.At the 6-mo follow-up,no specific symptoms were reported and dialysis was not considered.There were no unusual findings in the imaging study,and regular follow-up and imaging studies are scheduled.CONCLUSION This case report assessed the feasibility of simultaneous minimal invasive robotic surgery as an alternative to conventional open or laparoscopic surgery.

Synchronous primary carcinomas of the bladder and prostate

Aim： To determine the incidence of adenocarcinoma of the prostate for patients undergoing radical cystoprostatectomy for bladder cancer in Taiwan. Methods： A total of 248 patients in Taiwan who were histologically confirmed for transitional cell carcinoma of the bladder underwent cystoprostatectomy. Histopathologic evaluation of the prostate specimens sectioned at 5 mm intervals was performed. Results： Of the 248 patients, 10 （4.03%） were found to have prostate cancer. Of the 10 cases of unsuspected prostate cancer, eight proved to be at stage T1 or T2, and two at T3 and T4, respectively. This rate of incidentally found prostate cancer amongst our bladder cancer patients appeared to be lower than that found in bladder cancer patients in similar studies in USA. Conclusion： Although the incidence of incidental prostate cancer in patients in Taiwan with bladder cancer is not high compared with that in Western countries, we suggest that digital rectal examination and prostate-specific antigen （PSA） are important screening tools for men with bladder cancer, especially for those aged 60 years and older in Taiwan.

Retroperitoneal laparoscopic partial resection of the renal pelvis for urothelial carcinoma:A case report

BACKGROUND The standard treatment of transitional cell carcinoma of the upper urinary tract consists of radical nephroureterectomy with bladder cuff removal,which can be performed either in open or laparoscopy or robot-assisted laparoscopy.Treatment of chronic renal insufficiency patients with upper urothelial tumor is in a dilemma.Urologists weigh and consider the balance between tumor control and effective renal function preservation.European Association of Urology guidelines recommend that select patients may benefit from endoscopic treatment,but laparoscopic treatment is rarely reported.CASE SUMMARY In this case report,we describe a case of 79-year-old female diagnosed with urothelial carcinoma of the renal pelvis and adrenal adenoma with chronic renal insufficiency.The patient was treated with retroperitoneal laparoscopic partial resection of the renal pelvis and adrenal adenoma resection simultaneously.CONCLUSION Retroperitoneal laparoscopic partial resection of the renal pelvis is an effective surgical procedure for the treatment of urothelial carcinoma of the renal pelvis.

Upregulation of cell adhesion through delta Np63 silencing in human 5637 bladder cancer cells

Some researchs have demonstrated that the loss of delta Np63 is associated with aggressive phenotypes and poor prognosis. However, other research indicates that delta Np63 is considered to have oncogenic properties. Delta Np63 overexpression is often observed in association with the oncogenic growth of squamous cell carcinomas and bladder cancer. In this study, we investigated the oncogenic role of delta Np63 in regulating cell adhesion in transitional cell carcinoma of the bladder （TCCB）. The cells were stably transfected with the delta Np63 short hairpin RNA （shRNA） plasmid. Immunocytochemistry was performed to determine the knockdown efficiency. Tumour cells were studied for their ability to adhere to vascular endothelial cells. Confocal microscopy was used to analyse the changes in cytoskeletal F-actin. F-actin expression was measured by flow cytometry. Cell invasion ability was assessed using transwell chambers. The delta Np63-silenced tumour cells were shown to adhere more tightly than controls to vascular endothelial cells （P〈0.05）. The content of F-actin in the delta Np63-silenced cells was enhanced （P〈0.05）. The Matrigel invasion assays showed that human 5637 bladder cancer cells had a lower degree of motility when transfected with pdelta Np63-shRNA （P〈0.05）. In conclusion, silencing of the delta Np63 expression can enhance the adhesiveness of 5637 cells by inducing F-actin cytoskeleton production, and it will possibly inhibit the TCCB invasion and metastasis.

Single-cell transcriptomic analysis of tumor heterogeneity and intercellular networks in human urothelial carcinoma

Background:Heterogeneity of tumor cells and the tumor microenvironment(TME)is significantly associated with clinical outcomes and treatment responses in patients with urothelial carcinoma(UC).Comprehensive profiling of the cellular diversity and interactions between malignant cells and TME may clarify the mechanisms underlying UC progression and guide the development of novel therapies.This study aimed to extend our understanding of intra-tumoral heterogeneity and the immunosuppressive TME in UC and provide basic support for the development of novel UC therapies.Methods:Seven patients with UC were included who underwent curative surgery at our hospital between July 2020 and October 2020.We performed single-cell RNA sequencing(scRNA-seq)analysis in seven tumors with six matched adjacent normal tissues and integrated the results with two public scRNA-seq datasets.The functional properties and intercellular interactions between single cells were characterized,and the results were validated using multiplex immunofluorescence staining,flow cytometry,and bulk transcriptomic datasets.All statistical analyses were performed using the R package with two-sided tests.Wilcoxon-rank test,log-rank test,one-way analysis of variance test,and Pearson correlation analysis were used properly.Results:Unsupervised t-distributed stochastic neighbor embedding clustering analysis identified ten main cellular subclusters in urothelial tissues.Of them,seven urothelial subtypes were noted,and malignant urothelial cells were characterized with enhanced cellular proliferation and reduced immunogenicity.CD8^(+)T cell subclusters exhibited enhanced cellular cytotoxicity activities along with increased exhaustion signature in UC tissues,and the recruitment of CD4^(+)T regulatory cells was also increased in tumor tissues.Regarding myeloid cells,coordinated reprogramming of infiltrated neutrophils,M2-type polarized macrophages,and LAMP3^(+)dendritic cells contribute to immunosuppressive TME in UC tissues.Tumor tissues demonstrated enhanced angiogenesis mediated by KDR^(+)endothelial cells and RGS5^(+)/ACTA2^(+)pericytes.Through deconvolution analysis,we identified multiple cellular subtypes may influence the programmed death-ligand 1(PD-L1)immunotherapy response in patients with UC.Conclusion:Our scRNA-seq analysis clarified intra-tumoral heterogeneity and delineated the pro-tumoral and immunosuppressive microenvironment in UC tissues,which may provide novel therapeutic targets.

Techniques to resect the distal ureter in robotic/laparoscopic nephroureterectomy

Treatment of clinically-organ confined high grade urothelial carcinoma of the upper tract has historically comprised open nephroureterectomy,with the distal ureter and bladder cuff mobilized through a separate open pelvic incision.To decrease morbidity,urologists have increasingly adopted laparoscopy and robotics in performing nephroureterectomy.In many published series of laparoscopic nephroureterectomy,the distal ureter and bladder cuff are detached from the bladder endoscopically by a variation of the“pluck”technique,with the resulting bladder defect left to heal by prolonged indwelling urethral catheter drainage.While the distal ureter and bladder cuff can be excised laparoscopically,it does require advanced laparoscopic skills.With the wrist articulation and stereoscopic vision in robotic surgery,robotic nephroureterectomy(RNU)and bladder cuff excision can be performed in antegrade fashion to mimic the open technique together with the ability to intracorporeally close the bladder defect in a watertight,mucosa to mucosa fashion after excising the bladder cuff.In this review,we discuss the published minimally invasive techniques in resecting the distal ureter and bladder cuff during laparoscopic and RNU.