Objective To study the effects of emodin on apoptosis and mRNA and protein of apoptosis inducing factor (AIF) and Endonuclease G (Endo G) in human bladder cancer cells BIU87,and to investigate the anticancer mechanism of emodin. Methods The BIU87 cells were divided into 4 groups,control group,Z-VAD-FMK group,emodin group,emodin combined with Z-VAD-FMK group.The effects of different concentrations of emodin at different action time on cells proliferation of BIU87 in vitro culture were measured by methylthiazole (MTT) chromatometry,the cells apoptosis were detected by flow cytometry,and expression of AIF and Endo G were examined by reverse transcription PCR (RT-PCR) and Western blot.Results MTT assay demonstrated that the higher concentration of emodin and the longer action time,the more significant inhibition of tumor cell growth.Based on the IC50 value,80 μmol/L and 72 h of emodin intervention were selected as an intervention condition.The apoptosis rate in emodin group (44.57 ± 1.52 ) ％was significantly higher than that in emodin combined with Z-VAD-FMK group (35.58 ± 1.61 ) ％ ( P ＜0.01).RT-PCR and Western blot showed that the mRNA and protein of AIF in emodin combined with Z-VAD-FMK group,emodin group,control group,Z-VAD-FMK group were ( 1.74 ± 0.11 ) and (2.59 ±0.13),(1.36±0.08) and (1.89±0.14),(0.37 ±0.02) and (0.53±0.11),(0.42 ±0.06) and (0.44 ± 0.07),respectively.There were significant differences between emodin group and the other groups (P ＜0.01 ).The mRNA and protein of Endo G in emodin combined with Z-VAD-FMK group,emodin group,control group,Z-VAD-FMK group were (2.28±0.15) and (3.31 ±0.36),(1.85 ±0.13) and (2.15 ±0.27),(0.53 ±0.07) and (0.71 ±0.16),(0.61 ±0.04) and (0.67 ±0.22),respectively.The differences were significant between emodin group and the other groups ( P ＜ 0.01 ). Coneltusion Emodin can upgrade the expression of AIF and Endo G in bladder cancer cells BIU87,which can induce apoptosis through Caspase-independent pathway.

ObjectiveTo study the suppressive role of emodin on the growth and its effect on the proliferation cycle and apoptosis of human bladder cancer cell line BIU87.MethodsThe effect of different concentration of emodin at different time point on cell proliferation of BIU87 were measured with methylthiazole (MTT) chromatometry, the cell proliferation cycle were detected with flow cytometry, expressions of bc1-2 and caspase-3 were detected by SP of immunohistochemistry.ResultsWithin a certain range, the higher concentration of emodin (10 ～ 80 μg/ml) and the longer action time are positive related the more significant inhibition of tumor cell growth and the higher apoptosis rate [(9.84 ± 1.13)％, (18.32 ±2.14)％ ,(29.73+1.42)％ ,(42.13 +2.36)％ ,respectively].Compared with control group [(2.01 ±0.92)％], the differences were statistically significant(F =531.85, P ＜0.01).Emodin could inhibit the proliferation of human bladder cancer cell BIU87 by blocking BIU87 cell in G0/G1 stage, thus cut down cell proportion in stage of S [(33.27 +1.26)％ ,(29.17 ±1.39)％, (16.94 ±0.86)％ ,(10.85 ± 1.47)％,respectively], compared with the control group [(35.45 ± 0.38) ％], the differences were statistically significant(F =524.64, P ＜0.01).After 48 h of emodin treatment, the bc1-2 expression(Grayscale values:122.65 + 2.12,131.37 ± 1.62,134.81 ± 1.36,145.55 ± 2.01, respectively) was decreased and the caspase-3 expression(Grayscale values : 135.26 + 1.41,130.22 ± 1.74,126.11 ± 1.77,118.36 + 1.53, respectively) was increased in a dose dependent manner.Compared with control group (Grayseale values:108.42 + 3.73,149.35 ± 1.82, respectively), the differences were statistically significant (F = 216.23,224.83, P ＜0.01).ConclusionsEmodin could significantly inhibit the growth and induce apoptosis of BIU87 cells in vitro, which may be through down regulation of bc1-2, and up regulation of caspase-3, and blocking BIU87 cell in G0/G1 stage.

Objective To investigate the inhibitory effect of emodin on hetertransplanted human bladder cancer in nude mice and explore its mechanism.Methods Heterotransplanted models of human bladder cancer cell line BIU87 cells in nude mice were established.The mice were randomly divided into 4 groups during the experiment:blank control group,Z-VAD-FMK group,emodin group and emodin combined with Z-VAD-FMK group.The growth of tumors was observed and the growth curve was mapped.The nude mice were sacrificed 4 weeks later,the tumors were isolated and weighed.The pathological changes of tumor were observed after HE staining,the cells apoptosis were detected with flow cytometry,and the expression of AIF and Endo G were examined by reverse transcription PCR (RT-PCR) and Western blot.Results The tumor growth rate in emodin group was lower than that in the other three groups.The tumor quality in emodin group [(0.41 ± 0.05 ) g] and emodin combined with Z-VAD-FMK group [( 0.69 ±0.07)g]were lighter than that in the other two groups[(1.08 ±0.13,1.04 ±0.09)g,],and the differences were statistically significant( F =90.56,27.49,P ＜0.01 ).The quality difference in emodin group and emodin combined with Z-VAD-FMK group was statistically significant ( t =10.01,P ＜ 0.01 ).The apoptosis rate in emodin group [(42.71 ±2.69)％]was significantly higher than that in emodin combined with Z-VAD-FMK group[(34.38 ± 1.73)％] ( t =6.38,P ＜0.01 ).The expression of AIF and Endo G in emodin combined with Z-VAD-FMK group was significantly increased than other groups [( 1.65 ±0.12)vs(1.24±0.08),(0.51 ±0.07),(0.48 ±0.04);(2.12 ±0.16)vs(1.75 ±0.13),(0.57 ±0.06),(0.59±0.07);(2.42±0.13)vs(1.73 ±0.11),(0.78 ±0.07),(0.75 ±0.08);(3.13 ±0.25)vs(2.15± 0.18 ),(0.85 ± 0.09 ),(0.81 ± 0.14 )],and the differences were significant ( F =303.22,319.32,409.38,258.53,P ＜ 0.01 ).Conclusions Emodin could significantly inhibit the growth of hetertransplanted human bladder cancer in nude mice.The mechanism might be partly due to the expression increase of AIF and Endo G in bladder cancer cells,which might induce apoptosis through Caspase-independent pathway.

Objective　To analyze the effects of flexible ureteroscopic Holmium laser lithotripsy on renal function in the treatmen of renal calculi.　Methods　From October 2010 to March 2012,41 cases of solitary renal calculi (24 males and 17 females) were treated with flexible ureteroscopic Holmium laser lithotripsy ( FURL).Patient's mean age was 51.9 ± 15.9 years ( range from 31 to 88 years).Locations of renal calculi detected by image study were 22 cases in middle and upper calyx,9 cases in lower calyx,10 case in renal pelvis.The mean size of calculi was 16.9 ±6.0 mm (range from 10 to 28 mm).Blood samples (2 h pre-operatively,and then 2 h,12 h,24 h,48 h,72 h post-operatively) were collected and serum　NGAL,serum Cys-C were tested　Results　The measured levels of pre-operative NGAL and Cys-C were 3.5 ± 0.6μg/L,501.7±121.3 μg/L,and levels of post-operative NGAL at 2 h,12 h,24 h,48 h and 72 h were 4.2±0.8 μg/L,5.0±1.0 μg/L,4.9±1.4 μg/L,4.3± 1.1 μg/L and 3.8 ±0.1 μg/L,while the according levels of Cys-C were ( 516.4 ± 126.2 ) μg/L,( 723.8 ± 134.8 ) μg/L,( 770.4 ± 162.8 ) μg/L,(671.7 ± 138.3 ) μg/L and 574.0 ± 116.7 μg/L.Serum NGAL began to increase 2 hours after operation (P ＜0.05,and reached the peak in 12 hours,it began to decline 12 hours after surgery,but it was still higher at 72 hours than pre-operative level (P ＜ 0.05.Serum Cys-C showed no obvious ascent 2 hours after operation in FURL group ( P ＞ 0.05 ),but increased obviously 12 hours after operation and lowered down after the peak that occurred 24 hours after surgery.Serum Cys-C still remain above the baseline 72 hours after operation (P ＜0.05).　Conclusions　Flexible ureteroscope lithotripsy can cause reversible damage to renal function after surgery.

Objective To evaluate the efficiency and clinical value of modular flexible ureteroscope combined with Holmium laser lithotripsy for treatment of renal calculi. MethodsA total of 46 patients with renal calculi were treated with modular flexible ureteroscope combined with Ho:YAG lithotripsy in our hospital from December 2009 to May 2011.There were 19 cases in middle and upper calyx,8 cases in lower calyx,15 case in renal pelvic,and 4 cases in multiple calyxes.The calculi diameter ranged from 6mm to 31 mm (average in 15 ± 7.9 mm).Flexible ureteroscope was used to find renal calculus under epidural anesthesia,and Holmium laser lithotripsy was used at a maximum energy of 12 to 20W (0.8 - 1.0 J/15 -20Hz).F5 or F6 double-J tubes and catheters were indwelled routinely postoperatively.-examine with KUB and removethe catheter 2 days after operation.Re-examine with B-ultrasonography or KUB 4 weeks after op-eration to evaluate the stone free rate.Residual stones ≥4 mm were regarded as clinical significance of residual stone fragments. Results 45 operations were successful except one case changed to percutaneous nephrolithotomy (PCNL) because of ureteral stricture.The operation time was 45 -150 min (average in 110.4 ±25.3 min).The patients were discharged from hospital in 2 -5 days(average in 3 days) after operation.The stone detection rate was 95.6％ (44/46),and the stone free rate was 86.9％ (40/46) after one operation. No severe complications such as ureteral perforation or hemorrhea occurred. Conclusions Modular flexible ureteroscope combined with Holmium laser lithotripsy is effective and safe for treatment ofrenal calculi,meanwhile it can decrease the high maintenance costs of integrated flexible ureteroscope.

Objective To evaluate the efficiency and clinical value of modular flexible ureteroscope combined with Holmium laser litho-tripsy on the treatment for upper ureteral calculi of solitary kiden. Methods There were 26 cases of upper ureteral calculi of solitary kidney, including 12 cases of left ureter calculi and 14 cases of right ureter calculi. The diameter of the calculi ranged from 0. 6~2. 2 cm, averagely (1. 38 ± 0. 22) cm. F6 double-J tubes were indwelled postoperatively for 4 weeks, and catheters were indwelled postoperatively for one week. Examine with KUB one day after operation to evaluate the effective rate. Results The Ureteral guiding sheath were successfully implanted in 23 cases, 2 cases underwent second lithotripsy after indwelling D-J tubes 2 weeks later, D-J tubes failed to be implanted only in one case, which has changed to percutaneous nephrolithotomy (PCNL) for ureter stricture. The stone free rates was 88. 5%(23/26)after the first ses-sion,and it was 100%(26/26)at the end of the session. The operation time was 15~45 min, meanly (27 ± 3. 2) min. One patient had ureter perforation during the operation, and one patient had severe infection after operation. Conclusion The modular flexible ureteroscopy com-bined with Holmium laser lithotripsy is an effective and safe treatment, especially for upper ureteral calculi of solitary kideny, and it should be considered as the first choice for solitary kidney calculi.

Objective: To evaluate the success rate during ultrasound-guided percutaneous nephrolithotomy (PCNL) and analyze the reasons of puncture failure. Methods: A retrospective analysis was performed based on the data of 58 patients who underwent ultrasound-guided PCNL by 4 experienced urologists(10 years' experience of PCNL and more than 80 cases per year)in our center from August 2018 to November 2018. Of all the 58 patients, there were 36 males and 22 females (aged from 22 to 73 years) with the mean age of 51 years. The calculi ranged from 9 mm to 93 mm, with the average of 26.5 mm. The separation of renal collecting system ranged from 5 mm to 30 mm, with the average of 15.1 mm. All of the 58 patients underwent one-stage PCNL and the numbers of punctures, the numbers of percutaneous tubes and the reasons for failure were recorded. Results: All percutaneous tubes and surgeries were established and done successfully. Of all the 118 punctures, 74 punctures succeeded by detecting the urine and 68 surgery tracts were established (6 punctures failed because of the dilation). The total puncture success rate was 62.7%(74/118). Of the total 74 successful punctures, 56.8%(42/74)succeeded at the first puncture, 28.4%(21/74)succeeded at the second puncture and 14.9%(11/74)succeeded at least after three punctures. The success puncture rate of the 4 urologists were 59.2%(29/49), 64.1%(25/39), 66.7%(16/24), 66.7%(4/6)respectively, and there were no statistical differences between the puncture success rates of the urologists (P=0.679). Each channel needed 1.7 punctures on average. Fouty-four punctures failed without detecting the urine, including 20 failed punctures because of the puncture tract deviation, 17 punctures without seeing the urine after the core needle being removed, 7 punctures no display on the ultrasound imagine. There were 33 punctures to be tubeless while other 35 indwelling the nephrostomy tubes. Five nephrostomy tubes' position were different with the preoperationally predicted position which means the discrepancy rate was 14.3%(5/35). One patient had complications and classified as Grade II or above on the modified Clavien Grading System of aerothorax. Conclusions The puncture success rate during ultrasound-guided PCNL is not satisfied. The main reasons of puncture failure are the deviation of puncture tract, unclear imaging of puncture tract and other unclear reasons( puncture needle went too deep or superficially or tip of the needle embedded into the stone).

Objective To detect the feasibility and safety of applying all-seeing needle in transurethral seminal vesiculoscopy.Methods Retrospective analysis was made with clinical data of 32 patients of hemospermia treated with transurethral seminal vesiculoscopy using all-seeing needle from March 2016 to January 2018.The patients'age was (38.8 ± 8.7) years (27-60 years) and the course of disease was (7.1 ±3.3) months (2-15 months).Ultrasound before operation showed heterogeneous echo,or expansion of the seminal vesicle.MRI showed hemorrhage of the seminal vesicle,or abnormal signal of the seminal vesicle.Patients had levofloxacin or mosisasin anti-infection therapy more than one month and remained uncovered.The operation was performed under subarachnoid anesthesia,and the patients took the lithotomy position.The F4.8 all-seeing needle entered the posterior urethra,the verumontanum was found,and the saline was slowly pushed with a syringe to maintain a clear view.Then,the ejaculatory duct opening was searched on both sides of the verumontanum.If the ejaculatory duct opening cannot be found in the normal position,we entered the needle into the prostatic utricle to find the possible ectopic opening.If the ejaculatory duct opening was still not found,at the 5 and 7 o'clock positions in the prostatic utricle,the needle was probed and punctured into the side wall of the ejaculatory duct.Visible puncture with all-seeing needle can effectively avoid penetrating blood vessels and reduce damage to tissues during puncture.In this study,the ejaculatory duct opening got accessed on the verumontanum in 14 cases,through ectopic openings within the prostatic utricle in 2 cases,and through artificial establishment in 5 and 7 o'clock positions within prostatic utricle in 16 cases.After entering the ejaculatory duct and seminal vesicle,we explored the cavities of the seminal vesicles.For stones or polyps,after replacing the outer sheath to F8,F1.9 stone retrieval basket was applied to remove stones or polyps,followed by rinsing the seminal vesicles with normal saline,0.02％ nitrofurazone,and then 160,000 units of gentamicin into each seminal vesicle.For hemorrhage,after clearing up the blood,seminal vesicles were also washed with normal saline,nitrofurazone,and perfused with gentamicin.In the operation,prostatic utricle stone was found in 5 cases,and seminal vesicle stone was found in 7 cases.One case of seminal vesicle polyp was observed,and in 19 cases,seminal vesicle hemorrhage was seen.Results All the operations were accomplished successfully,the operation time was (55.0 ± 11.3) min (35-82 min).There was no rectal injury or urethral injury during operation.The postoperative catheter was removed at 2 days postoperatively and the patients discharged on the 4th day after surgery.The length of hospital stay was (6.3 ± 0.7) days (5-7 days).One patient had mild hematuria after removal of the urethral catheter and got improved spontaneously;one case developed epididymitis,which improved after anti-infective treatment.6 cases made stone analysis,with 5 cases of magnesium ammonium phosphate hexahydrate,and 1 case of calcium oxalate dihydrate,calcium oxalate monohydrate and carbonate apatite mixed stones.One case of polyp was diagnosed by pathologists to be an inflammatory polyp.Follow-up was performed at 4,6,8,and 12 weeks after surgery.Hemospermia was cured in 24 cases,relieved in the other 8 cases at 8-week follow-up,who received oral antibiotic treatment afterwards.At the 12-week follow-up,the rest 6 cases in 8 got hemospermia recovered,with 2 cases still uncovered.Conclusions All-seeing needle is user-friendly and safe in transurethral seminal vesiculoscopy,with reliable short-term efficacy.

Objective To observe the influence of ratio of endoscope-sheath diameter on renal pelvic pressure during PCNL.Methods The model of 24 isolated adult porcine kidneys were used to imitate percutaneous nephrolithotomy from September 2016 to June 2017.Each tract was established (F12,F14 and F16).Three kinds of endoscopes (F8 all-seeing needle percutaneous nephroscope,F6/7.5 and F8/ 9.8 rigid ureteroscope),input a 200μm laser,were adopted.There were 8 combinations,and 3 kidneys were used for each one.Renal pelvic pressure was measured 3 times repeatedly in every combination with steady irrigation (50,100,150,200,250,300,350,400,450,500 cmH2O).Results The linear equations,ratios of endoscope-sheath diameter and highest renal pelvic pressures for each combination were:F8-F12,Pr =0.026 P0-1.533,0.67,12.2 ± 0.54;F6/7.5-F 12,Pr =0.112 P0-5.001,0.92,51.2 ± 0.93;F8-F 14,Pr =0.010P0 + 1.067,0.58,6.2 ± 0.48;F6/7.5-F14,Pr =0.020P0 + 1.000,0.79,10.8 ± 0.46;F8/9.8-F14,Pr =0.144P0 +20.933,0.87,84.7 ± 1.17;F8-F16,Pr =0.005P0 + 1.067,0.50,2.8 ±0.34;F6/7.5-F16,Pr =0.009P0 + 0.533,0.68,5.1 ± 0.32;F8/9.8-F16,Pr =0.020P0 + 2.200,0.75,12.6 ± 0.56.The combinations of F6/7.5-F12 and F8/9.8-F14 might lead to a high renal pelvic pressure without proper irrigation (＜401.7 cmH2O for F6/7.5-F12;＜ 132.4 cmH2O for F8/9.8-F14).Conclusion In order to keep a safe pressure,a proper ratio of endoscope-sheath diameter (＜ 0.80) and appropriate irrigation must be considered.

Objective To discuss primary experience the clinical use of "All-seeing needle set" combined with Holmium laser to treat the single renal stone lesser than 2 cm in diameter through a super minimal tract (F12).Methods From January 2015 to December 2016,43 patients were enrolled into this retrospective study.There were 23 males and 20 females who were diagnosed as single renal stone less than 2 cm in diameter(age range from 23-65 years).There were 8 upper pole renal stones,13 lower pole renal stones and 22 renal pelvis stones.The mean stone size was (1.63 ± 0.32) cm in diameter (range from 1.2 cm to 2.0 cm)."All-seeing needle system" was applied during percutaneous puncture.After building a F12 minimal tract by fascia dilator,all patients received lithotripsy with Holmium laser.Indwelling catheters for 3 days and an F6 double-J tube was left for 2 weeks without a tube in the percutaneous tract after operation.Finally,KUB and/or urinary CT were used to check the results on day 1.Result All operations were completed successfully.The operation time was 23-65min [averaged (31.0 ±9.2) min].41 cases' renal stones were store free,and the 2 stone residues was clear with later extracorporeal shock wave lithotripsy.No severe complications occurred.Conclusion As an alternative to standard procedures for treatment of renal stones less than 2 cm in diameter,adopting "All-seeing needle set" with Holmium laser through a super minimal tract (F12) is safe,minimally invasive,fast and effective with a low complication rate.