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Int. braz. j. urol ; 45(6): 1161-1166, Nov.-Dec. 2019. tab
Article in English | LILACS | ID: biblio-1056350


ABSTRACT Purpose: To compare two-shift operation mode and single player mode different impact on surgical results and operator comfort in flexible ureteroscopic holmium laser lithotripsy for renal calculi larger than 1.5cm. Materials and Methods: From december 2017 to december 2018, 92 patients with renal calculi admitted to Qilu Hospital and were treated through flexible ureteroscopy. They were randomized in two-shift group (n=50) and single player group (n=42). The operative time, blood loss, hospitalization stay after operation, residual fragments (≥4mm) rate, fragmentation speed, postoperative complications and operator's fatigue score were compared. Results: There was no significant difference between two groups regarding age, gender, illness side, stone size, blood loss, operative time, postoperative hospitalization stay, complications, etc (p >0.05). The fragmentation speed was 44.5±20.0mm3/min in two-shift group compared with 34.2±17.3mm3/min in single player group (p=0.037). Residual fragments (≥4mm) rate after first surgery was 18% in two-shift group, while the residual fragments (≥4mm) rate was 40.5% after first surgery in single player group (p=0.017). The total fatigue score of two-shift group was 8.4 compared to 29.9 in single player group (p <0.001). Conclusion: In flexible ureteroscopic holmium laser lithotripsy for the treatment of renal calculi larger than 1.5cm, two-shift operation mode can raise the fragmentation speed and stone clearance rate, as well as significantly lower operator's fatigue level and improve operator's comfort.

Humans , Male , Female , Adult , Kidney Calculi/surgery , Lithotripsy, Laser/methods , Ureteroscopy/methods , Lasers, Solid-State/therapeutic use , Postoperative Complications , Kidney Calculi/pathology , Reproducibility of Results , Treatment Outcome , Lithotripsy, Laser/instrumentation , Ureteroscopy/instrumentation , Equipment Design , Operative Time , Ergonomics , Length of Stay , Middle Aged
Chinese Journal of Urology ; (12): 386-390, 2010.
Article in Chinese | WPRIM | ID: wpr-389276


Objective To obtain shRNA sequences that can stably block the expression of Nuclear Factor kappa- B (p65) in the prostate cancer cell line LNCaP and construct the lentivirus vector.And validate the gene function of p65 in the cell line. Methods According to p65 genetic information, we design siRNA1, siRNA2, siRNA3 those three siRNA sequences targeting the ods area of p65 gene and then form the corresponding four pairs of complementary single strand DNA of shRNA, including the sense strand and the antisense strand. The synthetic shRNA sequence was inserted into the empty pSIH1-H1-copGFP shRNA Vector, and after transfecting the prostate cancer cells , the inhibitory effect of p65 mRNA by different sequences was detected through real-time PCR, and the inhibitory effect of p65 protein expression was detected by Western-blotting. Thus we can obtain highly effective shRNA sequences in the inhibition of p65 in prostate cancer cells. MTT, flow cytometry, transwell were chosen to test the cell growth, migration and invasive power in vitro to compare the difference of the experimental group, control group and negative group. Results The third shRNA sequence had the best inhibitory effect and the inhibitory effect of p65 mRNA in prostate cancer cell line was 59 % and the protein was 81%. It's position locates in p65 (NM_021975 ) 1096-1113 and it's stemloop sequence is 5'-GATCCGCCCTATCCCTTTACGTCATTCAAGAGATGACGTAAAGGGATAGGGCTTTTTG-3'. After transfecting, the prostate cancer cell line had the low expression of p65 stably. Through MTT, we got the growth curve, which showed that the growth ability of experimental group was significantly decreased compared with the control group and the Logarithmic growth didn't appear in the first 96 hours. Flow cytometry test displayed that the percentage of G0-G1-phase cells in experimental group was 61.49%, and the control group was 44.89%, idle group was 41.52%, which was increasing oberviously. The S-phase cells in the experimental group was 28.58%, compared with the 47.36% and 46. 10% diminished. The results of transwell showed that the experimental group had 16. 5000±6. 62076 cells and the other two groups had 45. 6333 13. 54159 and 36. 8333±5. 68412 cells, which showed the invasive power of experimental group was significantly declined(P<0.05).Conclusions It's successful to obtain shRNA sequences that can stably block the expression of p65 in the prostate cancer cell line LNCaP and construct the lentivirus vector. p65 can positively regulates the biological behavior of prostate cancer LNCaP cell line in the cell growth, migration and invasive power.

Chinese Journal of Urology ; (12): 561-564, 2010.
Article in Chinese | WPRIM | ID: wpr-387618


Objective To verify the efficiency of a new laparoscopic urethrovesical anastomosis training model by comparing it with the chicken skin model. Methods Chicken posterior trunks and porcine colons were used to construct the training model. The posterior trunk of a chicken was used to simulate a human pelvis, and a 3-mm cloacal stump was used to simulate a human urethral stump. A 15-cm segment of porcine colon with a 1-cm orifice was used to simulate a human bladder or neobladder. An imitation urethrovesical anastomosis was performed with laparoscopic instruments in a laparoscopic training box. Forty trainees with no laparoscopic experience were randomized into 2 groups.The trainees in group A (n=20) practiced using this new model for 8 h, while those in group B (n=20) practiced using the chicken skin model for 8 h. The trainees' skills were assessed using the porcine model before and after training. Results Compared with the chicken skin model, this new training model more accurately resembled the structure and characteristic of human pelvis, urethral stump, and bladder (neobladder). After the training sessions, both groups improved in anastomosis time [GroupA: (64±11)min vs. (123±20)min, P<0.05; Group B: (77±12)min vs. (121±17)min, P<0.05] and quality (Group A: 8.8± 1.0 vs. 3.8 ± 1.2, P<0. 05 ; Group B: 7.7 ± 0.9 vs.3. 7± 1.1, P<0. 05). Compared with trainees in group B, trainees in group A required less time and achieved a higher quality score (P<0.05). Conclusions This new training model can help urologic surgeons to reduce learning curve of this technique and improve their suturing skills. It is an effective,convenient training model for laparoscopic urethrovesical anastomosis.

Article in Chinese | WPRIM | ID: wpr-528278


AIM: To identify the non-steroid transcription factors upregulating the expression of L-plastin in hormone-independent prostate cancer, and partly elucidate the mechanism of hormone-refractory prostate cancer. METHODS: TF SEARCH software was used to analysis the possible binding sites of transcription factors in the 3’ end of L-plastin promoter that had been identified as important part of regulation response elements. Gel shift assay and supershift assay were used to confirm the transcription factors binding the speculated response elements. PCR site-mutagenesis technique was performed to delete the binding site of transcription factor and luciferase activity assay was carried out after deletion of the binding site. RESULTS: SP-1 respond element GGTGGGGCGGGGA located at -54- -41 of L-plastin promoter was identified with the TF SEARCH software. Gel shift assay and supershift assay confirmed that SP-1 was the transcription factor binding to GGTGGGGCGGGGA. Mutant deleted the SP-1 binding-site had low-luciferase activity than that of the naive. CONCLUSION: SP-1 plays an important role in the up-regulation of L-plastin expression in hormone-independent prostate cancer.

Article in Chinese | WPRIM | ID: wpr-529133


AIM: To construct eukaryotic expression vector of small interfering RNA(siRNA) specific to bcl-2 and investigate the effect of recombinant plasmid on suppressing bladder cancer cell growth.METHODS: siRNA of bcl-2 gene was designed according to the principle of RNAi-based medicine, and was converted into cDNA coding expression of small hairpin RNAs(shRNA) of siRNA. The cDNA was synthesized and inserted into plasmid pGenesil-1. The recombinant eukaryotic expression vectors of pGenesil-1545 and pGenesil-1555 were controlled by the U6 promoter of RNA polymerase Ⅲ, identified by the restriction map and the sequence analysis, and transfected into T24 cells. After T24 cells were transfected for 72 h, expression of bcl-2 mRNA was assayed by RT-PCR; and MTT was used to observe the proliferation of T24 cells.RESULTS: The recombinant plasmids of pGenesil-1545 and pGenesil-1555 were identified by the restriction map and the sequence analysis. The sequences completely coincided with the designs. The expression of the bcl-2 mRNA in T24 cells transfected with recombinant plasmid decreased nearly 80%, and the growth of T24 cells was suppressed significantly.CONCLUSION: The siRNA eukaryotic expression vector against bcl-2 gene is successfully constructed. It effectively downregulates the expression of bcl-2 in T24 cells and suppresses the cell growth.