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【Objective】 To explore the correlation between CSAG1 expression and the prognosis and tumor-infiltrating lymphocytes in renal clear cell carcinoma (RCCC), and to predict the survival and tumor progression. 【Methods】 The gene expression profiles and clinical information of CSAG1 were downloaded from the Cancer Genome Atlas (TCGA). Based on the differential mRNA expression, GO annotation and KEGG pathway analysis were performed. The relationship between CSAG1 and tumor immune infiltration was assessed with Tumor Immunoassay Resource (Timer 2.0) database. The mRNA expression of CSAG1 in human RCCC specimens was validated with qRT-PCR. 【Results】 CSAG1 expression was significantly higher in RCCC tissues than in normal tissues (P<0.05). The qRT-PCR results revealed that the mRNA level of CSAG1 was consistent with that predicted by bioinformatic analysis. The KEGG analysis and GO annotation indicated high GSAG1 expression in RCCC was related to transmembrane transport, tricarboxylic acid cycle and lysosome. CSAG1 expression was positively related to the infiltration of pDC, aDC, CD8+ T cells, cytotoxic cells, TFH, TH1 cells, Tem, NK CD56dm cells, Treg and T cells, but negatively correlated with macrophage infiltration. 【Conclusion】 CSAG1 may be associated with poor prognosis of RCCC and become a potential immunotherapy target.
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【Objective】 To investigate the efficacy and safety of single position transabdominal and extraperitoneal laparoscopic radical nephroureterectomy in the treatment of upper tract urothelial carcinoma (UTUC). 【Methods】 Clinical data of 31 UTUC cases treated in our hospital during Nov.2018 and Jun.2022 were retrospectively analyzed, including 11 tumors in the right side, and 20 in left side. There were 14 cases of renal pelvic carcinoma, 16 cases of ureter carcinoma, and 1 case of renal pelvic carcinoma plus ureter carcinoma. 【Results】 All surgeries were successfully performed without conversion to open surgery. The mean operation time was (81.45±19.80) min, and the estimated blood loss was (69.03±24.13) mL. No serious perioperative complications were observed. The average postoperative hospital stay was (6.13±2.44) d, and the median follow-up was 28 (3.0-49.0) months. At the last follow-up, 2 patients died, 3 had recurrence, but no contralateral recurrence was observed. 【Conclusion】 Single position transabdominal and extraperitoneal laparoscopic radical nephroureterectomy is safe, effective and feasible in the treatment of UTUC. It is worth clinical popularization.
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【Objective】 To explore the safety and efficacy of a modified one-piece posterior laparoscopic total nephroureterectomy with cystic sleeve resection in the treatment of upper urinary tract uroepithelial carcinoma (UTUC). 【Methods】 A total of 24 patients treated during Jan. and Jun. 2022 were involved, including 16 males and 8 females, aged 62 to 90 (average 73) years. The UTUC was in the left side in 15 cases, and in the right side in 9 cases. There were 10 cases of renal pelvis tumor, 6 cases of upper ureteral tumor and 8 cases of lower ureteral tumor. 【Results】 All operations were successful without conversion to open surgery. The operation time ranged from 60 to 100 minutes, average (71.25±9.80) minutes. The intraoperative bleeding volume was 20 to 200 mL, average (30.03±8.13) mL. No significant intraoperative or postoperative complications occurred. The postoperative hospital stay was 4 to 7 days, average (5.83±1.44) days. Bladder perfusion chemotherapy was performed after surgery. 【Conclusion】 The modified one-piece posterior laparoscopic total nephroureterectomy plus cystic sleeve resection for UTUC is an effective and feasible procedure with satisfactory tumor control, which is worth further promotion in clinical practice.
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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.
Subject(s)
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 AgedABSTRACT
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.
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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.
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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.
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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.