Objective To study the effect of Numb gene overexpression on invasion and migration in human renal cell carcinoma cells and its related mechanism. Methods The Numb-ORF plasmid was transfected into renal cell carcinoma cells 786-O, set the negative control and blank control. The expression of Numb and matrix metalloproteinase-9 (MMP-9) was detected by real-time PCR and western blot. Then abilities of cells invasion and migration were measured respectively by transwell assay and scratch-wound-healing assay. Results Compared to negative control (2.51±0.27) and blank control (2.87±0.21), Numb expression in Numb-ORF group (36.13 ±8.33) was significantly higher (P= 0.00). Meanwhile, MMP-9 level in Numb-ORF group was decreased: Numb-ORF group was 2.36±0.29, negative control was 8.73±0.91, blank control was 8.99±0.78 (P=0.00). In scratch-wound-healing assay,the repair ratio of 12 h was as follow: Numb-ORF group was 0.53 ±0.06, negative control was 0.73 ±0.09, blank control was 0.75 ±0.08 (P= 0.02). In transwell assay, the number of invasion cells in Numb-ORF group was 31.40±5.96, less than negative control (126.93±13.61) and blank control (131.87 ±15.42) (P= 0.00). Conclusion Overexpression of Numb significantly decreases abilities of invasion and migration in 786-O cells, and the suppressive effect may be due to the down-regulation of MMP-9 expression in human renal cell carcinoma.

Objective To study the effect of Numb overexpression on invasion and migration in human bladder cancer cell line T24,and its related mechanism.Methods In June 2013,the plasmids including Numb-ORF plasmid and its blank vector pCMV6-Entry were transfected into T24 cells,and set the blank control as usual.The expressions of Numb and MMP-9 (Matrix metalloproteinase-9) were detected by Real-Time PCR and Western Blot.Then T24 cell invasion and migration were measured respectively by Transwell assay and Scratch Wound Healing Assay.Results Compared to negative control group,which transfected with blank-vector plasmid and untransfected group,Numb expression level in Numb-ORF group was significantly higher (P＜0.01).Meanwhile,MMP-9 level in Numb-ORF group was decreased,△ CT-value as follows:Numb-ORF group was 8.423±0.202,negative control group was 7.294±0.138,untransfected group was 7.220±0.118 (P＜0.01).In Scratch Wound Healing Assay,the repair ratio of Numb-ORF group was less than negative control group and untransfected group.The repair ratio in 12 hours as follow:Numb-ORF group was 0.525±0.037,negative control group was 0.693±0.034,untransfected group was 0.701 ±0.038(P=0.004).In Transwell assay,the number of invasion cells in Numb-ORF group was 12.6±3.2,less than in control group (130.8±9.2) and untransfected group (132.2± 10.6) (P＜0.05).Conclusions Overexpression of Numb by transfection significantly decreases the capability of T24 cell invasion and migration.This suppressed effect may be due to the down-regulation of MMP-9 expression in human bladder cancer.

Objective To study the effect of Numb gene on CXCR4 expression and proliferation and migration of bladder cancer. Methods Human bladder cancer cells were divided into 3 groups:experimental group(transfected with Numb-ORF plasmid),negative-control group( transfected with blank vector),and blank-control group(no DNA transfected in). The expressions of Numb and CXCR4 were detected by real-time PCR and Western blotting respectively. Cell proliferation and migration were measured by MTS and Transwell assay respectively. Results Numb expressions in experimental group,negative-control group and blank-control group were 31. 044 ± 3. 350,4. 401 ± 0. 567 and 4. 287 ± 0. 341 respectively,with a statistical significance (F = 183. 418,P = 0. 000). CXCR4 levels in experimental group,negative-control group and blank-control group were 0. 344 ± 0. 167,0. 996 ± 0. 148 and 1. 010 ± 0. 106 respectively,with a statistical significance (F = 21. 355,P = 0. 002). In MTS assay,the absorb values in experimental group,negative-control group and blank-control group were 0. 615 ± 0. 057,0. 987 ± 0. 063 and 0. 957 ± 0. 066,with a statistical difference(F =33. 210,P = 0. 001). In Transwell assay,the numbers of migratory cells in experimental group,negative-con-trol group and blank-control group were 164. 667 ± 19. 858,670. 133 ± 38. 760 and 667. 533 ± 27. 610,with a statistical significance(F = 286. 788,P = 0. 000). Conclusion Overexpression of Numb can suppress the ability of proliferation and migration of the BIU-87 cells through down-regulation of CXCR4 expression in human bladder cancer.

Objective To assess the clinical efficacy of a modified complete retroperitoneal laparoscopic nephroureterectomy via 3 port approach.Methods From August 2013 to February 2016,23 patients with complete retroperitoneal laparoscopic renal and ureteral sleeve resection were treated with modified three port approach,including 15 males and 8 females.The average age was 67 years old (ranging 44-83 years old).All patients had complained about the hematuria before operation and urine exfoliated cells showed moderate to severe nuclear atypia.All patients accepted the abdominal CT and urography CTU examination,pre-operatively.All of them was diagnosed localized upper urinary tract malignant tumors based on those images,including 13 cases in the pelvis,and 10 cases in the upper segment of the ureter.No chemotherapy,radiotherapy or immunotherapy was performed before surgery.No patients have the history of severe basic disease or upper urinary procedure.The operations were performed under general anesthesia,patients take the contralateral back 30 degrees slope,low elevation head foot,waist bridge,side waist stretch.In the anterior superior iliac spine perpendicular to the line 2 cm parallel to the lower intersection of the rib border were disposed into the 12 mm trocar.Above the anterior superior iliac spine two cross finger level with the intersection of the anterior axillary node,we placed into the 10 mm trocar placement lens.Laparoscopic placement of third casing form an isosceles triangle with the first two casing.The renal fascia was incised with an ultrasonic knife from the renal dorsal side,and the renal hilum was isolated from the kidney by suction aspirator.The renal artery and vein were separated and closed by hem-o-lok.Along the psoas muscle surface to ureter,ureteral clipping by hem-o-lok but not to cut off the free distal ureteral,the lens is composed of first casing into,using ultrasonic knife to free ureter to the bladder wall segment,with 30 mm endoscopic stapler ureter and bladder wall cut off part.Operation time,blood loss and postoperative recovery were recorded in 23 cases.Results All 23 cases were successfully operated without related the operative complication.The operative duration ranged from 3.5 to 6.1 h (mean 4.8 h),the blood loss was 30-880 ml (mean 304 ml),and the postoperative stay was 8-30 d (mean 17.8 d).There are 3 cases of positive lymph node by postoperative pathological reports.Within 2 to 30 months following up,2 patients died of tumor progression in 6 months after surgery.4 patients were diagnosed with bladder cancer in 15 months,15 months,21 months,24 months after surgery,respectively.And the transurethral resection of bladder tumor was performed.Conclusion The modified complete retroperitoneal laparoscopic nephroureterectomy via three ports is safe and reliable.

Objective To investigate the effect of Notch1 gene dowuregulated on migration and proliferation of human prostate cancer cell line PC-3.Methods The small interfering RNA (siRNA) targeted Notch1 gene or negative control sequences was transfected into PC-3 cells.The expression of Notch1 or Hes1 gene was detected by Real-Time PCR and Western Blot.Then ability of migration or proliferation was measured by Transwell assay or MTS Assay.Results Compared with negative control group (36.097±1.941) and untransfected group (38.762±1.897),Notch1 expression level in siRNA group (3.960±0.510) was significantly reduced (P ＜ 0.01).Meanwhile,Hes1 level in siRNA group was decreased,expression in three groups as follows:siRNA group was 1.690±0.994,negative control group was 8.776±0.916,untransfected group was 9.803±1.001 (P ＜ 0.01).In Transwell assay,the number of migration cells in siRNA group was 657.867±27.610,more than that in the negative control group (158.533±18.263) and untransfected group (146.933±15.733) (P ＜ 0.01).In MTS assay,there was no significant difference among three groups at 0 h point,however,siRNA group was significantly raised at the time points of 24,48 and 72 h (P ＜ 0.01).Conclusions Downregulation of Notch1 gene by transfection of the siRNA-Notch1 sequences significantly promoted ability of migration or proliferation in PC-3 cells,and the effect may be due to the down-regulation of Hes1 expression.

Objective To study the effect of Numb gene on cell cycle and proliferation in human renal carcinoma cells and its related mechanism. Methods Renal carcinoma cells Caki-1 were taken as research objects, and the Numb-ORF plasmid transfected cells, negative control group and blank control group were set respectively. The expression levels of Numb and cyclin D1 were detected by real-time PCR and Western blot. The cell cycle was analyzed respectively by flow cytometry. Cell proliferation was assessed by comparing with absorbance at 490 nm using a micro-plate reader. Results Compared to the negative control group (5.05±0.45) and blank control group (5.13±0.31), the △Ct value of Numb in the Numb-ORF group (1.92±0.39) was significantly lower. The protein levels in the Numb-ORF group, the negative control group and the blank control group were 6.67±0.83, 3.08±0.47, 2.85±0.36, respectively (P = 0.00). Meanwhile, the △Ct value of cyclin D1 in the Numb-ORF group (6.20±0.87) was higher than that in the negative control group (4.35±0.51) and the blank control group (4.56±0.31) (P = 0.02), and the protein level in the Numb-ORF group (5.85±0.72) was lower than that in the negative control group (10.04±0.83) and the blank control group (11.88±1.26) (P = 0.00). The ratio of G0/G1 cells in the Numb-ORF group was (54.29±4.15) %, the negative control group was (38.69±2.60) % and the blank control group was (41.28±1.29) % (P = 0.00). In proliferation assay, compared with the negative control group (0.93±0.10) and the blank control group (1.02±0.06), the A value at 24 h in the Numb-ORF group (0.67±0.07) was significantly reduced (P = 0.00). Conclusion Numb gene could increase the cell percentage in G0/G1 phase and inhibit proliferation of renal carcinoma cells via down-regulating the expression of cyclin D1.

Objective To observe the effect of a novel pressure controlled water jet used in transurethral resection of bladder tumor. Methods Clinical data of 12 patients who underwent a transurethral resection of bladder tumor by using a novel water jet was retrospectively analyzed. Results All 12 cases were successfully operated. Estimated blood loss during operation was less than 5 ml. Operation time was 20-45 minutes (median was 28 min), time of water injection was 8-15 minutes (median was 12 min). There were no significant complications. All patients were followed up for 4-10 months, and tumor recurrence or progression was not found by cystoscopic examination. Conclusion Transurethral resection of bladder tumor by using a novel pressure controlled water jet is advantageous with favorable safety and feasibility.

Objective To evaluate the efficacy and economic results in the treatment of upper ureteral calculi by surgery of ureteroscopic Holmium laser lithotripsy under the supine half-sitting lithotomy position.Methods To analyze the clinical data of upper ureter calculi patients who underwent ureteroscopy Holmium laser lithotripsy retrospectively.Patients in traditional lithotomy position with assistance of stone basket during the surgery were in group A.Those in traditional position surgery without stone basket were in group B.Those in supine,half-sitting,lithotomy position and without the assistance of stone basket were in group C.There were 62,31 and 75 patients in group A,B and C respectively.In group A,there were 32 males and 30 females;37 cases in left side,25 in right side,with degree of hydronephrosis of (2.9 ±0.6) cm and stone size of (1.7 ±0.4) cm.In group B,there were 18 males and 13 females;14 cases in left side,17 in right side,with degree of hydronephrosis of (2.8 ± 0.6) cm and stone size of (1.6 ±0.5) cm.In group C,there were 44 males and 31 females;36 cases in left side,39 in right side,with degree of hydronephrosis of (2.8 ± 0.7) cm and stone size of (1.7 ± 0.5) cm.There was no significant difference in aspects of age,degree of hydronephrosis and stone maximum diameter.Statistical analysis was performed regarding operation time,fee for hospitalization and stone free rate among the three groups.Results Ittook (71.8±9.6) min,(62.2±ll.4) min and (65.4±6.8) min in group A,BandC respectively,and there was significant difference among three groups.When comparing two groups respectively,the operation time in group A was more than group B and C significantly,while group B and C shared no difference.As for fees in hospitalization,they were (27.2 ± 4.0),(22.4 ± 5.0) and (22.4 ±3.8) thousand Yuan in three groups,and significant difference was found.By comparing two groups,we found that the fees in group A were more than group B and C significantly,while group B and C had no difference.Finally,stone free rate was compared among three groups,they were 90.3％,64.5％,96％ respectively,and there was significant difference as well.By further study,we found that stone free rate in group C was the highest,followed by A and B group.Conclusions There were higher stone free rates,less operation time and expense to perform ureteroscopic Holmium laser lithotripsy in supine half-sitting lithotomy position.