Objective To describe our renal sinus anatomy based tension-free reconstruction technique step by step and report perioperative data and long-term outcomes of patients undergoing robotassisted nephron-sparing surgery for hilar tumors.Methods From June 2013 to December 2016,data of 286 consecutive patients with hilar tumor who underwent RAPN in single center were retrospectively reviewed.There were 202 males and 84 females,aged (56.2 ± 9.2) years.The body mass index was (26.8 ± 3.5) kg/m2.The median diameter of tumor was 2.6 cm(0.8-6.0 cm),and R.E.N.A.L.score was 8.2 ± 1.8.The anatomy-based "Garland" technique specialized in protecting the large hilar vessels and minimizing the tension of trans/retroperitoneal defect suturing approach for anterior/posterior lip hilar tumor respectively.Patient's perioperative complications and long-term follow-up including renal function and oncological outcomes were analyzed.Results "Garland technique" was successfully applied in 284 patients,the warm ischemia time (WIT) was (18.2 ±4.1) min.Median estimated blood loss (EBL) for RAPN was 100 ml (range:10-600 ml).Median operative time was 120 min (range:60-230 min).No patient was converted to open surgery.Postoperative hospital stay was 4.0 d (range:2.0-9.0 d).Three patients (1.1％) had positive surgical margins.Of all the pathological results,260 cases (91.5％)were clear renal cell carcinoma,8 cases (2.8％)were chromophobe renal carcinoma,7 cases (2.5％)were papillary type renal cell carcinoma,5 cases(1.8％) were oncocytoma,3 cases (1.1％)were angiomyolipoma,one case (0.3％) was mucinous tubular and spindle cell carcinoma.Two patients underwent blood transfusion.Three patients(1.0％) had local recurrence.284 patients were alive at a median follow-up of 36 months (range:12-54 months).Conclusions "Garland technique" is safe and feasible for hilar tumor resection and reconstruction with less surgical complications.Large renal vessel injury was avoided and tension of wound closure was minimized.The trans/retroperitoneal approaches are capable for anterior/posterior hilar tumor.Patients with hilar tumor could benefit from robotic surgery with a well preserved renal function and good oncological outcomes.

Objective To investigate the effect of acupuncture combined with human amniotic mesenchymal stem cells ( HAMCs ) transplantation on the osteoporosis in ovariectomized rats.Methods Forty-five healthy female non-pregnant Wistar rats were randomly divided into sham operated group, model group and experimental group ( acupuncture+HAMC transplantation), 15 rats in each group.At 7 days after the surgical wound healing, the rats received acupuncture every day and intravenous injection of HAMCs suspension ( except the sham operated group) once a week for consecutive 12 weeks.The body weight was measured every week.24 hours after the last intervention, the rats were killed and specimens were collected.Specimens of the mid-, proximal-and distal femur were taken to measure the bone mineral density using a dual energy X-ray absorptiometer.At 12 weeks after the intervention, changes of the expression of serum 25-hydroxy-vitamin D (25-HVD), C-terminal peptide of type I collagen (CTX-I), bone specific alkaline phosphatase ( BAP) and human tartrate-resistant acid phosphatase 5b ( TRACP 5b) were determined by ELISA, the biomechanical properties of the removed femur was measured, and the expression of transforming growth factor-β( TGF-β) in the vertebrae was assessed by RT-PCR assay.Results Body weight of rats in the sham operation group was increased gradually in accordance with the natural growth of animals, that of the model group was significantly higher than that of the sham operated group since two months after modeling, and the body weight of the experimental group was similar to that of the sham operated group.The bone mineral density and calcium content of the model group were significantly decreased compared with that of the sham operated group ( P<0.05) , and the bone mineral density and bone calcium content of the experimental group were significantly higher than that of the model group ( P <0.05 ) .The ELISA showed that the expressions of serum 25-HVD, CTX I, BAP, and TRAP5b in the model group were significantly lower than that of the sham operated group, and those of the experimental group were significantly higher than that of the model group ( P<0.05 ) . Measurement of biomechanical properties showed that the limit load, limit stress and elastic modulus of the femur of the sham operated group were significantly higher than that of the model group ( P<0.05 ) , and the limit load and elastic modulus of the experimental group were significantly higher than that of the sham operated group ( P <0.05 ) .T-PCR showed that the expression of TGF-β1 m-RNA in the experimental group was significantly up-regulated than that of the sham operated and model groups ( P<0.05) .Conclusions Acupuncture combined with human amniotic mesenchymal stem cell ( HAMCs) transplantation has a synergistic effect on the treatment of osteoporosis, and can improve the conditions in ovariectomized rats.

Objective To evaluate the efficacy of self-retaining suture (QuillTM SRS) in retroperitoneal laparoscopic partial nephrectomy for complicated renal tumor by assessing perioperative parameters.Methods Between 2010 and 2012,78 cases of complicated renal tumor (R.E.N.A.L score ≥ 7) treated by retroperitoneal laparoscopic partial nephrectomy (LPN) with two layers continuous knotless barbed suture (QuillTM SRS group) (n=30) or traditional absorbable vicyl suture (non-SRS group) (n=48) were retrospectively analyzed.In QuillTM SRS group,2-0 Quill SRS was used to suture the deep wound bed,and the second outcr layer renorrhaphy was performed with a 1-0 Quill SRS by the same way.In non-SRS group,the inner layer was sutured using a 15cm in length 2-0 monicryl suture by the same method mentioned above.A second outer layer was sutured with 1-0 vicryl suture across the wound.Cases were matched for R.E.N.A.L score.Comparison was made in term of operation time,preoperative parameter and perioperative complications between SRS group and non-SRS group.Results Renorrhaphy was successfully performed in all cases except 1 case converting to open surgery in non-SRS group.Mean warm ischemia time in SRS group was shorter than non-SRS group (18 vs 25 min,P =0.021).The proportion of bleeding requiring intervention in the non-SRS group (7/48,14.5％) was 4.3-fold higher than that of the SRS group (1/30,3.3％),but the differernce is not significant (P＞0.05).There were no significant differences between two groups in postoperative creatinine changes.Limitations of this study include the absence of randomization and the relative small sample size.Conclusions SRS can be safely used for complicated renal tumor during LPN,and SRS can significantly reduce the WIT and may also reduce bleeding during the operation.

MicroRNAs (miRNAs or miRs) are a class of short, non-coding RNAs that participate in various oncological processes. This study aims to explore the roles of microRNA-34a (miR-34a) in invasive urothelial bladder carcinoma. miR-34a was transfected into bladder cancer cell lines 253J and J82. The miR-34a expression levels in tissues and cells were detected by using qRT-PCR. The Notch1 expression was detected by qRT-PCR and Western blotting. Cell migratory and invasive abilities were measured by Transwell chamber assay. Bioinformatics and luciferase assay were performed to predict and analyze the binding sites between miRNA-34a and Notch1. It was found that there was aberrant expression of miR-34a in bladder cancer tissues. Moreover, we revealed that ectopic expression of miR-34a suppressed cell migration and invasion, while forced expression of Notch1 increased cell migratory and invasive abilities. Finally, we observed that miR-34a transfection significantly down-regulated luciferase activity and reduced the mRNA and protein levels of Notch1. Our study concluded that microRNA-34a antagonizes Notch1 and inhibits cell migration and invasion of bladder cancer cells, which indicates the tumor-suppressive function of microRNA-34a in bladder cancer.
Adult ; Aged ; Cell Movement ; genetics ; Down-Regulation ; genetics ; Female ; Gene Targeting ; Humans ; Male ; MicroRNAs ; genetics ; Middle Aged ; Neoplasm Invasiveness ; Receptor, Notch1 ; physiology ; Transfection ; Tumor Cells, Cultured ; Urinary Bladder Neoplasms ; pathology ; physiopathology

MicroRNAs (miRNAs or miRs) are a class of short, non-coding RNAs that participate in various oncological processes. This study aims to explore the roles of microRNA-34a (miR-34a) in invasive urothelial bladder carcinoma. miR-34a was transfected into bladder cancer cell lines 253J and J82. The miR-34a expression levels in tissues and cells were detected by using qRT-PCR. The Notch1 expression was detected by qRT-PCR and Western blotting. Cell migratory and invasive abilities were measured by Transwell chamber assay. Bioinformatics and luciferase assay were performed to predict and analyze the binding sites between miRNA-34a and Notch1. It was found that there was aberrant expression of miR-34a in bladder cancer tissues. Moreover, we revealed that ectopic expression of miR-34a suppressed cell migration and invasion, while forced expression of Notch1 increased cell migratory and invasive abilities. Finally, we observed that miR-34a transfection significantly down-regulated luciferase activity and reduced the mRNA and protein levels of Notch1. Our study concluded that microRNA-34a antagonizes Notch1 and inhibits cell migration and invasion of bladder cancer cells, which indicates the tumor-suppressive function of microRNA-34a in bladder cancer.

ObjectiveTo investigate the effects of over expression of miR-34a on cellular proliferation and migration in bladder cancer cell line J82 by targeting Notchl.MethodsmiR-34a was predicted as a putative gene which can target Notchl through bioinformatics analysis,qRT-PCR and Western blot were performed to measure the expression levels of Notchl and miR-34a in invasive transitional cell carcinoma of bladder (TCCB) tissues and J82 cells transfected with miR-34a.Luciferase assay was employed to determine if miR-34a could target Notchl through binding to the 3'-untranslated region (3'UTR) of Notchl mRNA.J82 cells were transfected with pcDNA3.0-miR-34a or pcDNA3.0 control plasmid.MTS colorimetry was used to evaluate the effect of miR-34a on cell proliferation.The effect of miR-34a on cell migration was assessed by transwell migration assay.ResultsThe expression level of miR-34 in invasive TCCB tissues was lower than in adjacent bladder tissues (0.016(0.018) vs 0.042 (0.059),N =16; P =0.0006).On the contrary,the average levels of Notchl mRNA and protein were higher in tumors than in adjacent bladder tissues (2.765(2.156) vs 2.312(1.365),N =16; P =0.0025 and 0.857 ±0.197 vs 0.648 ±0.171 ;P ＜0.0001 ).After the transfection of miR-34a,the expressive level of miR-34a in J82 was highly induced ( (2.408 ±0.789) × 10-4 vs(0.153 ±0.029) × 10-4; P =0.0026).However,the expressive levels of Notchl mRNA and protein were obviously decreased (3.001 ± 0.106 vs 4.998 ± 1.053 ; P =0.0308 and 0.747 ± 0.050 vs 0.988 ± 0.102 ; P =0.0215 ).The results of luciferase assay showed that firefly activity was highly dimished (0.422 ± 0.028 vs 2.392 ± 0.148 ; P ＜ 0.0001 ).Cellular proliferation was inhibited after the transfection of miR-34a in J82 (P ＜ 0.0001 ).Moreover,number of migration cells of J82 was significantly reduced after the ectopic expression of miR-34a ( 179.3 ± 21.02 vs 269.7 ± 23.71 ; P =0.0078 ).ConclusionsmiR-34a inhibits the cellular proliferation and migration of bladder cancer cell line J82 via binding to the 3UTR of Notchl mRNA.

Objective To study the value of the preoperative detrusor contractility to the outcome assessment of prostatectomy for benign prostatic hyperplasia (BPH).Methods A total of 109 patients with BPH were analyzed.Their ages ranged from 62 to 83 years with a mean of 71 years.All patients underwent urodynamic study to confirm a diagnosis of BOO preoperatively.Further more, their BOO was not caused by nervous, endocrine or other diseases.Pateints were divided into two groups based on maximum detrusor contractility.Group Ⅰ (n =61, BPH with maximum detrusor contractility ≥ 40 cm H2O, 1cm H2O =0.098 kPa) underwent TURP or open surgery, respectively.Group Ⅱ (n =48, BPH with maximum detrusor contractility ≤ 20 cm H2O ) underwent TURP and suprapubic punctural cystostomy simultaneously,the bladder fistula was kept open continuously for at least two weeks postoperatively.The difference in outcome between the two grous was assessed by using urodynamic parameters including maximum detrusor contractility, Qmax and residual urine at one and three months postoperatively respectively.Student's t-test was used to compare the result for normally distributed data and Wilcoxon's signed-ranks test for skewed data in this study.Results There was significant difference in preoperative maximum contractility, Qmax between group Ⅰand groupⅡ (78.4 ±37.0 cm H2O) vs (19.2 ±5.4 cm H2O)(P＜0.01), (7.6±2.2 ml/s) vs (2.5 ± 1.1 ) ml/s (P ＜ 0.05) respectively.Although there was significant difference at one month postoperatively in Qmax (17.4 ±2.9)ml/s vs (12.5 ±2.0)ml/s (P＜0.05), no significant difference was found in Qmax between the two groups after three months ( 18.3 ±2.8 ml/s) vs ( 15.2 ± 1.8)ml/s (P ＞ 0.05).Conclusions The Qmax may improve and the impaired detrusor recovered gradually after the BOO was removed.Performing an operation on patients with BOO accompanied with detrusor underactivity may be useful to recover detrusor contractility.

Objective To study the influence of inhibited steroidogenic factor-1 on human adrenocortical H295R cells, and explore its role in the pathogenesis of adrenal tumors. Methods The plasmids pGenesil1-SF-1-shRNA which containing U6 promoter and SF-1-specific short hairpin RNA (shRNA) and pGenesil1-negative-shRNA containing unspecific shRNA were transfected into H295R cell. The expression of SF-1 was measured by Western blot and real-time polymerase chain reaction(RT-PCR). Cell proliferation was analyzed by WST-1 assay and cell count. Ki-67 expression was detected by immunohistochemistry and cell apoptosis was examined by TUNEL assay. Results Compared with those in control cells, the protein and mRNA level of SF-1- transfected cells were reduced by 69.7% and 71.2% (P＜0. 01). WST-1 and cell count method showed that SF-1 gene silencing obviously inhibited cell proliferation(P＜0. 01). By contrast, there was a 3. 7-fold increase in the percentage of apoptotic H295R cells in SF-1-inhibited group than that of control group (P＜0. 01). Immunohistochemistry showed that Ki-67 positive cells in SF-1-inhibited cells were lower than the negative control cells (16.90±2.17) % and (33. 48±3.16)%,(P＜0. 01). Conclusion SF-1 gene silencing can inhibit the proliferation of adrenocortical cells, and it is expected to become a key protein in understanding pathogenesis of adrenal tumors or treating them.

Objective To develop and evaluate a porcine model for training the single needle running suture method of laparoscopie urethrovesical anastomosis(LUA). Methods Twenty minipigs with mean weight of 30kg were general anaesthetized with Sumianxin solution 0. 1 ml/kg intramuscularly. Pneumoperitoneum was created by insufflation of carbon dioxide by a veress needle inserted through the umbilicus. One 10mm port and two 5mm ports were positioned after the establishment of pneumoperitoneum. The intestine was used as "bladder". The procedures were completed with the single needle running suture method of laparoscopic urethrovesical anastomosis. Six trainees performed the LUA procedure based on the models during a laparoscopic training course, following the technique used in the operation room. The learning curve was analyzed by operative time. Results The porcine model for laparoscopic training was established successfully and 3 LUAs could be performed on each pig. Each trainee performed 10 LUAs based on the models during the training course of laparoscopic urology. The operative time declined from (55.3±10. 4)min initially to (22.4±4.8)min (P＜0. 01) after the training course. At the end of training, all trainees could accomplish a watertight LUR procedure on the model. Conclusions The establishment of the training model is feasible. The trainees could acquire the skills necessary to perform LUA in vivo based on this model. The model provides a platform for training the basic techniques of LUA procedures.

Objective To study the role of fibulin-5 in urothelial carcinoma of bladder. Methods Fibulin-5 expression was detected in bladder cancer tissues (13 cases of G_1 and G_2, 7 cases of G_3) and normal bladder mucosa samples by Western blotting assay. Fibulin-5 cDNA was amplified by PCR and cloned into pMD-19T simple vector. The pMD-19T-Fibulin-5 vector was digested by restriction endonucleases XhoI and EcoRI to generate a XhoI-Fibulin5-EcoRI fragment that was then ligated into the identical sites in p-EGFP-Nl plasmid to synthesize p-EGFP-Fibulin-5 plasmid. The p-EGFP-Fibulin-5 plasmid was finally transfected into bladder cancer cell line 5637. The migration and invasion of untransfected, vector-transfected and fibulin-5-transfected bladder cancer cells were measured by Boyden chamber assay. Results Compared to 1. 16 ±0. 28 in the normal control, the expression of fibulin-5 protein in low grade and high grade tumors were 0. 57±0. 32 and 0. 44±0. 42(P<0. 01, respectively). However, the difference between low grade and high grade tumors was not statistically significant (P>0. 05). The successfully transfected bladder cancer cells demonstrated green fluorescent light. The migrated cell number of fibulin-5-transfected cells was 127. 6 ± 3. 1 compared with 139. 3±7. 7 for vector-transfected cells and 136. 9±5. 7 for untransfected cells (P>0. 05, respectively). In contrast, the invaded cell number of fibulin-5-transfected cells was 8. 0±3. 1 compared with 31. 5±4. 8 for vector-transfected cells and 31. 7±4. 7 for untransfected cells (P<0. 01, respectively). Conclusion Fibulin-5 is down-regulated in urothelial carcinoma of bladder and acts as a tumor suppressor gene by inhibiting the invasion of bladder cancer cells.