Objective:To investigate the influence of anoxia-reoxygenation on the transcription of TGF?1 gene in Human umbilical veins endothelial cell line(ECV304). Methods:Anoxia-reoxygenation model of Human umbilical veins endothelial cell line was established. TGF?1 mRNA was half-quantitatively analyzed by a modified RT-PCR. Results:The transcription of TGF?1 mRNA was up-regulated significantly in anoxia-reoxygenation group compared with the control group. Conclusion: anoxia-reoxygenation may up-regulate the transcription of TGF?1 gene in human umbilical veins endothelial cell line.

Objective:To construct short hairpin RNA(shRNA)eukaryotic expression vector targetinghuman TGF-?1 gene,and evaluate its inhibitory effects on TGF-?1 gene in endothelial cell lin(eECV304)of human umbilical veins in order to select a high efficacy vector for subsequent RNAi study.Methods:The oligonucleotides targeting TGF-?1 mRNA were designed and synthesized according to bioinformatics methods.The annealed duplex strand were cloned into PGenesil-1 plasmid.Two recombinant plasmids(pT11 and pT12) were verified by enzyme digestion and sequencing,and transfected into endothelial cell line of human umbilical veins.Vector with scramble sequence was set as control.The mRNA expression was measured by semi-quantitative reverse transcriptase-polymerase chain reaction(RT-PCR)and the protein was measured by ELISA.Results:The shRNA expressing vectors targeting TGF-?1 gene were confirmed by SalI enzyme digestion and sequencing.The transfection rate of pT11 and pT12 in ECV304 were 38.2%and 40.1%,respectively.The inhibitory rate of TGF-?1 mRNA were 58.1%and 60.0%,and the inhibitory rate of TGF-?1 protein were 38.9%and 44.3%.Conclusions:The successfully constructed shRNA eukaryotic expression vectors specific for human TGF-?1 gene have significant inhibitory effects on the expression of TGF-?1 mRNA and protein in endothelial cell line of human umbilical veins,and the pT12 is more effective.

Objective:To investigate the correlation between overall recurrence rate,survival rate,and the first recurrence duration and operative methods,and to discuss the strategy of bladder-preserving for patients with bladder transitional cell carcinoma(TCC).Methods:A total of 249 cases of bladder tumor with complete follow-up data,treated from 1991 to 2005 in our hospital,were studied re trospectively.The correlation between overall recurrence rate,survival rate and the first recurrence duration and operative methods were analyzed by the life table method,and statistical methods,like Newman-Keuls and chi-square test.Result:The first recurrence duration was not influenced by operative methods. There was no significant difference of overall recurrence rate and short-term survival rate(3-year and 5-year)among different operative methods,the highest 10-year survival rate(77.3%)occurred in the group of transurethral resection of bladder tumor(TURB)and the lowest one(65.2%)in the group of partial cystectomy,and the difference was statistically significant.Conclusions:TURB could achieve the same short-term therapeutic effect as opening operations(partial resection of bladder or resection of bladder tumors)and the higher long-term survival rate can be obtained by TURB.

Objective To discuss the bladder-preserving strategy for bladder tumor.Methods Totally 249 cases of bladder tumor were studied retrospectively.The analysis of overall recurrence rate,survival rate and the first recurrence-free duration according to operative methods,drugs for therapeutic bladder perfusion,and clinical stages was carried out.Results The first recurrence-free duration was not influenced by operative methods and perfusion drugs significantly.Although the overall recurrence rate and short-term survival rate were not influenced by operative methods,but the highest 10-year survival rate(77.3%) occurred in the patients received transurethral resection of bladder tumor(TURB).The recurrence rate,3-year,5-year and 10-year survival rate of the patients applied to different perfusion drug were statistically different,with the lowest recurrence rate (28.2%) in patients applied Bacillus Calmette-Guerin(BCG) and the highest survival rate was in patients applied BCG(3-year: 100%,10-year: 81.7%).According to clinical stage,the survival rate was of no statistical difference among patients belonging to T_(is),T_a,T_1 stages,and of significant difference between T_(2-4) and the other three stages.The overall recurrence rate were not significantly different among T_(is),T_a and T_(2-4) stages.Conclusion TURB combined with BCG perfusion produces favorable prognosis for bladder tumor patients without muscle-involved,while bladder-preserving operation is not beneficial for patients with muscle-involved bladder tumor.

Objective To evaluate the modified POSSUM surgical scoring system in urologic surgery patients over 70. Methods 180 patients over 70 were retrospectively analyzed with the mo dified POSSUM surgical scoring system.A comparison between the observed and the predicted morbidity was made. Results In the complication group and noncomplication group,the mean age of the patients were 74.9?4.1 and 74.7?3.6,the mean length of stay were (42.1?4.2)d and (28.7? 14.2 )d,respectively.In the complication group,both the physiological scores and the operative severity scores of POSSUM scoring system were significantly higher than the no complication group ( 22.0 ? 4.1 vs 20.1?3.9 in pysiological score and 11.7?4.3 vs 10.3?3.5 in operative severity score, P

BACKGROUND: Many methods can reduce panel reactive antibody (PRA) levels in highly sensitized adult patients waiting for renal transplantation. Desensitisation with plasmapheresis or immunoabsorption is commonly used. Howevel the adverse effects limit their applications. OBJECTIVE: To investigate the effect of pretreatment of intravenous immunoglobulin (IVIG) on PRA levels in highly sensitized adult patients waiting for renal transplantation. DESIGN, TIME AND SETTING: Case analysis, self-contrast of 57 patients waiting for renal transplantation from Ren Transplantation Center. First Affiliated Hospital of Chongqing Medical University between January 2003 and Novembem 2006. PARTICIPANTS: Fifty-seven adult patients, who were highly sensitized to the human leucocyte antigen (HLA), agcd 21 to 65 years. were selected. The average baselines of PRA levels of patients were(46.7±29.5)%. METHODS: All patients received IVIG pretreatments at a dose of 5 g/d for 2 weeks. as one course of treatment. A weel later. some of them received additional 2 weeks iniection of IVIG. The PRA levels of patients were measured before and just after the IVIG treatments. MAIN OUTCOME MEASURES: PRA levels. RESULTS: It was significantly decreased below the non-sensitive level(＜10%)in 48 candidates(84.2%),and partially decreased in 6 ones(10.6%),however, was not changed in 3 ones(5.2%).The total effective rate was 94.8%.Thereafter.each candidate received a cadaveric renal graft. The preoperative complement-dependent cytotoxicity of each candidate was blow 10%. Furthermore. no hyperacute rejection was observed in all 57 recipients. Only two recipients experienced acute reiection, however, that was successfully conversed by methylprednisolone and FK506 treatments. CONCLUSION: Pretreatment of IVIG is a promising and effective option for lower allosensitization and can improve transplantation in highly sensitized candidates with end-stage renal diseases.

Objective To investigate the effect of antisense oligonucleotides (ASODN) targeting livin on the inhibition of livin mRNA and protein expression and the apoptosis of human renal carcinoma cell line 786-O cells. Methods Specific phosphorothioate antisense oligodeoxynucleotides targeting livin were synthesized and then transfected into 786-O cells. The expressions of livin mRNA were detected by RT-PCR. Expression and location of livin protein were observed by confocal laser scanning microscope (CLSM). Apoptosis rate of 786-O cells was investigated by flow cytometer. The activity of Caspase-3 was detected by colorimetric assay. Results After the transfection of ASODN, the expression of livin mRNA was decreased (P

With the deepening of the reform of public hospitals at the county level,as well as the development of medical technology personnel training and medical technology development in county hospitals,county hospitals should be able to make use of the latest technology to undertake the common diseases and frequently occurring diseases of county residents.So at present,the teaching hospitals affiliated to medical universities should carry out the short-term training of the latest special techniques in the training of the regular doctors.Department of Urology of the first Affiliated Hospital of Chongqing Medical University,in the 3 month training cycle,with improving the self-learning ability of the refresher doctors as the core,carried out the hand by hand teaching of small classes and introducing model teaching,which achieved good results.

Objective To explore the feasibility of detecting the cytokeratin 20 (CK-20) mRNA in exfoliated urothelial cells for the diagnosis of bladder carcinoma. Methods Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of CK-20 mRNA in cells collected from the urine of 45 cases of bladder cancer, 15 cases of cystitis accompanied by hematuria, 10 healthy volunteers, and 7 different cell lines, including bladder cancer cell line T24, kidney cancer 786-0 and GRC-1, breast cancer MCF-7 and MDA-MB-435, and ovary cancer SKOV 3 and 3AO. Results CK-20 mRNA expression was detected in 36 of 41 cases of bladder transitional cell carcinoma (87.80%), in 18 of the 21 GⅠ patients (85.71%), in 11 of the 13 GⅡ patients (84.62%), in 7 of the 7 GⅢ patients (100%), in 20 of the 22 T a-1 patients (90.91%), and in 16 of the 19T 2-4 patients (84.21%). Sensitivity of the method was found to be 87.80%, whereas specificity was 73.33%. In 15 patients with hematuria, there were 4 cases of false positive: 1 case of BPH, 1 case of atypical hyperplasia, 1 case of chronic inflammation, and 1 case undergoing TURP previously. CK-20 amplification band was also obtained in all of 19 cases of bladder transitional cell tumor tissues and bladder cancer cell line T24, but not in 4 patients with non-transitional cell carcinoma and 6 other tumor cell lines. No false positive cases were found in the healthy control group. Conclusion These results suggest that CK-20 might be a useful tumor marker for early noninvasive diagnosis and follow-up of bladder cancer by detecting CK-20 mRNA expression of uroepithelial cells from the voided urine specimen by RT-PCR.

The oncogenic isoform of the p63 protein, delta NP63, plays an important role in the pathogenesis of many epithelial carcinomas, and emerging evidences suggest that delta NP63 is a promising drug target. However, the functions of delta NP63 in transitional cell carcinoma of bladder (TCCB) are poorly defined. In this study, a delta NP63 shRNA expression vector was transfected into TCCB cell line 5637 and cell cycling, cell proliferation and protein expression were assessed by flow cytometry and 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-dimethyl tetrazolium bromide (MTT) assay, and immunohistochemistry, respectively. The delta NP63 shRNA expression vector was also injected into 5637 cell xenograft tumors in nude mice, and tumor size was measured, tumor tissue morphology was assessed by immunohistopathology and transmission electron microscopy. In the in vitro study, delta NP63 shRNA transfection caused successful delta NP63 gene silencing and resulted in significant arrest of cell cycling and cellular proliferation (p<0.05) as well as cyclin D1 expression. In the nude mouse xenograft model, delta NP63 shRNA greatly inhibited tumor growth, induced tumor cell apoptosis (p<0.05) and resulted in cyclin D1 downregulation. Our data suggest that delta NP63 may play an oncogenic role in TCCB progression through promoting cell survival and proliferation. Intratumoral administration of delta NP63-specific shRNA suppressed tumor delta NP63 expression and cellular proliferation while promoted tumor cellular apoptosis, and therefore inhibited tumor growth and improved survival of xenograft-bearing mice, which was not accompanied by significant signs of systemic toxicity.
Animals ; Carcinoma, Transitional Cell/*genetics/metabolism ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1/biosynthesis ; Disease Progression ; Female ; Humans ; Mice ; Mice, Nude ; Microscopy, Electron, Transmission ; Models, Biological ; Neoplasm Transplantation ; Trans-Activators/*biosynthesis/*physiology ; Tumor Suppressor Proteins/*biosynthesis/*physiology ; Urinary Bladder Neoplasms/*genetics/metabolism