Objective To study the effects of knock down midkine(MK)by siRNA on chemosinsitivity in bladder cancer cells. Methods Three MK siRNAs were designed and constructed. After transfected with MK siRNA or scrambled siRNA for different time, cultured cells were harvested to carry on the next experiments. Expression of MK was determined by real-time RT-PCR and western blot, and apoptosis were evaluated by caspase-3 activity and TUNEL assay. MTT was performed to examine the inhibition effect of Paclitaxel (PTX) on cells. Results MK siRNA could down-regulate the MK expression in a dose- and time-dependent manner. The MTT results showed that the inhibit rates were (18.21±0.36)%, (18.19±0.29)%, (17.89±0.33)%, (1.86±0.52)%, (32.56±0.53) %, (53. 83±0.38) % and (78. 95±0.55) % in different groups PTX alone(0.2μmol/L), ConA +PTX(0.2 μmol/L), Con-B +PTX(0.2 μmol/L), siRNA alone(12. 50 nmol/L), siRNA(3. 125nmol/L) + PTX (0. 2 μmol/L), siRNA (6. 25 nmol/L) + PTX (0. 2 μmol/L) and siRNA (12. 50nmol/L)+PTX(0.2 μmol/L), respectively. The TUNEL results showed that apoptosis index was (1.81 ±0. 36)%, (1. 89±0. 38)%, (5. 56±0. 58)%, (9. 68±0.55)% and (15. 25±0.56)% in different groups (Con-A, Con-B, siRNA (3. 125 nmol/L), siRNA (6. 25 nmol/L) and siRNA ( 12. 5nmol/L), respectively. The activity of caspase-3 increased significantly in transefected cells with a dose-and time-dependent manner. Conclusion MK siRNA could sensitize human bladder cancer cells to chemotherapy which might be through the apotosis.

Objective To analyze the effect of cyclosporin A(CsA), rapamycin(RPM) and macophenolic acid(MPA) on the co-stimulated lymphocytes, CD28 and CD40, and their production of Th1/Th2 cytokine, IL-2, IFN-γ, IL-4, IL-10 and IL-12. Methods The experimental groups were divided into ①mono-stimulating and co-stimulating groups: CD3 mAb mono-stimulating (group a),CD3 mAb+CD28 mAb co-stimulating (group b), CD3 mAb+CD28 mAb+CD40 L mAb co-stimulating(group c), CD3 mAb+CD28 mAb+CTLA4 mAb co-stimulating (group d). ②CsA groups: 300 ng/ml of CsA was added to group a, b, c and d. ③RPM groups: 300 ng/ml of RPM was added to group a,b, c and d. ④MPA groups:300 ng/ml of MPA was added to group a, b, c and d. The cytokine production was measured by ELISA.Results The co-stimulated CD28 and CD40 Th1/Th2 cytokines production of IFN-γ, IL-2, IL-4 and IL-10 were significantly increased. Compared with group a, IFN-γ increased from (248.91±11.20)ng/ml to (555.08±24.42)ng/ml and (548.19±33.06)ng/ml, IL-2 increased from (29.48±8.61)ng/ml to (1100.82±99.29)ng/ml and (842.23±29.31)ng/ml, IL-4 increased from (32.29±6.76)ng/ml to (116.02±15.03)ng/ml and (147.28±18.07)ng/ml, IL-10 increased from (147.01±10.47)ng/ml to (291.79±12.47)ng/ml and (302.52±35.18)ng/ml,respectively, P＜0. 01. Compared group b with group c, the Th1 cytokines production was decreased.IL-2 and IL-12 decreased (P＜0.05). The Th2 cytokine IL-4 production was increased (P＜0. 05).CTLA4 mAb and three other immunosuppressants, CsA, RPM and MPA, inhibited co-stimulated lymphocyte's both cytokines Th1 and Th2 production. The inhibitory effect of CsA on Th1/Th2 cytokine production was more significant than RPM and MPA did. The co-inhibitory effect of CTLA4 mAb and CsA was observed as well. The increased co-stimulated CD28 and CD40 IL-12 production could be suppressed by MPA. CsA and RPM had no inhibitory effect on the IL-12 production.Conclusions CD28/CD40 co-stimulatory pathway plays the key role in lymphocyte activation and Th1/Th2 cytokine production. CsA, RPM and MPA can inhibit co-stimulated lymphocyte's Th1 and Th2 cytokine production. CsA and CTLA4 mAb have co-inhibitory effect on co-stimulated lymphocyte's Th1/Th2 cytokines production. CD40 L mAb decreases the Th1 cytokines production(including IL-12) and increases the Th2 (mainly IL-4) production, which may be the mechanism of its longevity effect on allograft.

Objective To explore the mechanisms of protective effect of benazepril in the treatment of experimental cyclosporin- induced chronic nephrotoxicity. Methods Rats were on low-salt diet and cyclosporine A (CsA) was administered by gastric gavage at a dose of 30?mg/kg once daily for 28 days. The expression of mRNA for intrarenal transforming growth factor-?1 (TGF-?1) and renin was detected by reverse transcription-polymerase chain reaction (RT-PCR). Intrarenal expression of TGF-?1 and Collagen Ⅳ was determined by immunohistochemistry. The effects of benazepril on these changes were also evaluated. Results Chronic cyclosporine-induced nephropathy may be related to TGF-?1 and renin mRNA up-regulation as well as matrix proteins accumulation in interstitium. Benazepril could reduce TGF-?1 mRNA up- regulation and decrease intrarenal matrix proteins accumulation. Conclusion Decreased CsA-related TGF-?1 up-regulation expression and accumulation of matrix proteins in the kidney may be related to mechanisms of protective effect of benazepril in the treatment of cyclosporin-induced chronic nephrotoxicity.

Objective To analyze the characteristics of iatrogenic urerteral injury and summarize the experiences in prevention,diagnosis and treatment of iatrogenic urerteral injury. MethodsA review was made on the injurycauses,the injury locations,the treatment time,the methods of surgical procedures and the results of treatment in 17 patients with iatrogenic ureteral injury treated surgically from 1997 to 2003. Results Of 17 cases of iatrogenic ureteral injuries,gynecological,general surgical and urological procedures resulted in ureteral injuries in 12 cases (71%),four (24%) and one (6%),respectively. Of all the injuries,65% (11/17) appeared in the lower part of the ureter,18% (3/17) in the middle part of the ureter and 18% (3/17) in the upper part of the ureter. The main injury causes were ligation,partial ligation,complete transection and perforation,accounting for 29% (5/17),41% (7/17),24% (4/17) and 6% (1/17),respectively. Four cases were found during operation,nine at days 2-11 after operation and four were treated 3-6 months after injury. Treatment methods included end-to-end ureteral anastomosis in seven cases,ureteroneocystostomy in three,ureteral lithotomy in one,pure ureteral lysis in three and post-lysis double-J tube insertion in three. All patients were cured. The follow-up ranging from six months to three years showed no patients suffering from urinary tract infection,hydronephrosis or atrophy. Conclusions The location and type of injury determine the type of surgical repair. A thorough knowledge of pelvic anatomy and mastering the basic steps of diagnosis and treatment are critical for prevention and management of the iatrogenic urerteral injury.

Objective To explore the single point target concentration for Neoral at 2 h postdose (C2) in Chinese renal transplantation recipients for the first 3 months following surgical procedures. Methods Neoral trough levels (C0) and C2 monitoring were measured by fluorescence polarization immunoassay (TDX) in 114 cases of cadaver renal transplants treated with Neoral (6～7 mg?kg -1 ?d -1 ), mycopherolate mofetil (MMF,1.0～1.5 g/d) and steroids for the first 3 months after renal transplantation.The effectiveness of the new monitoring method in predicting the acute rejection and side effects was retrospectively analyzed. Results The acute rejection rate of 114 transplants for the first 3 months was 15.8%(18/114).The incidence of side effects was 30.7% (35/114),including hepatoxicity(26.3%) and nephrotoxicity(7.0%).The results of 234 pairs of Neoral C0, C2 monitoring showed that the difference was not statistically significant between C0 levels of rejection and non rejection,while the difference between C2 levels [(921.55 ?431.31) vs (1 185.17?358.86)ng/ml) ] was significant.There was also no statistically significant between C0 levels of the recipients with side effects and those without side effects,but statistically significant difference was found between C2 levels [(1 302.59?450.21) vs (1 105.23? 371.64 )ng/ml].Analysis of the relationships between C2 levels and the incidences of acute rejection and side effects showed that no acute rejection and side effects rate of 4.3% were observed in the Neoral C2 interval from 1 250 ng/ml to 1 500 ng/ml. Conclusions Neoral C2 monitoring is a more sensitive predictor not only for acute rejection but also for side effect rate.The optimal C2 target level of Chinese renal transplantation recipients is 1 250 ～1 500 ng/ml for the first 3 months post transplantation.

Objective To determine whether altered expression of apoptosis pathway genes is related to different apoptosis susceptibility of prostate cancer cells. Methods Androgen sensitive and insensitive prostate cancer cell lines LNCaP and PC 3 were cultured and treated by etoposide,and apoptosis were determined using Hoechst 33258 staining.The cells were harvested and the total RNA was extracted.cDNA probes were prepared and labeled with biotin 16 dUTP,then hybridized to commercially available cDNA arrays including apoptosis pathway specific genes. Results Apoptosis were induced in both PC 3 and LNCaP cells by etoposide,however,PC 3 was more resistant than LNCaP.Compared with LNCaP cells,the 4 fold down regulated genes in PC 3 cells were Bcl10,CIDE A,GADD45a,RIP2,caspase 4,5 and 6,while the 4 fold up regulated gene in PC 3 cells was TRAF4.Caspase 14 and TNFR2 were most strongly expressed genes in the 2 cell lines. Conclusions The altered expressions of apoptosis pathway specific genes are related to the different apoptosis susceptibility,and this may make an important contribution to androgen insensitive state of prostate cancer cells.

Objective To investigate the effect of murine dentritic cells vaccine modified with IFN-? inducible protein-10 gene on CTL induction. Methods DC was propagated from bone marrow (BM) of mice and pulsed with mouse prostate cancer cell line RM-1's whole lysate (Tuly-DC).IP-10 DNA fragments were inserted into pcDNA3.1(+) vector to construct recombinant plasmid IP-10/pcDNA3.1.Tuly-DC was transfected with IP-10/pcDNA3.1 by DOTAP liposome.Reversal transcript-polymerase chain reaction (RT-PCR) was used to evaluate gene transfer efficiency and chemotaxis assay was used to estimate the tansfected DC's chemotactic activity on T cells.Antitumor activity of the DC vaccine was studied in vitro by using Mixed leukocyte reaction (MLR) and cytotoxic assay (MTT assay). Results The IP-10 plasmid vector was successfully transfected into DC,which was confirmed by RT-PCR.The DC tranfected with IP-10 gene was capable of synthesizing and secreting IP-10 chemokine,which could increase the preferential chemotaxis of DC to T cells.MLR showed that the DC pulsed with whole tumor lysate and modified with IP-10 gene (IP-10/ Tuly-DC) could induce T cell proliferation significantly compared with other groups (P

Objective To evaluate the efficacy and safety of Simulect for the prevention of acute rejection in renal allograft recipients receiving Neoral-based immunosuppressive regimen. Methods A prospective,multicenter and open-label clinical trial was conducted from March to October 2001.A total of 33 patients [20 men and 13 women; age range,18-63 years;mean age,(42.6?11.6) years] who received first kidney allograft were enrolled.Thirty-two cases had panel-reactive antibody

Objective To investigate the feasibility of reconstruction of ureter with tubularized peritoneal free grafts for the treatment of avulsion of ureteral mucosa using animal models.Methods Twelve adult dogs were randomly divided into the reconstruction group(n=6) and the control group(n=6).Firstly,the model of avulsion of ureteral mucosa about 3-5 cm long was made.In reconstruction group,tubularized peritoneal free grafts and ureteral stents were placed in the injured ureters;and in control group,no operation was performed but to place the stents into the ureter.The curative effect was observed by IVU and histological examination 10 weeks after operation.Results In reconstruction group,IVU showed normal size and morphology of the kidneys.There was no hydronephrosis,and no obvious stricture of the part of ureter in which peritoneal free grafts were used as mucosa substitutes.In control group,IVU showed no image or only obscure enlarged outlines of the kidneys,and no image of the ureters.Atresia or severe stricture of the ureters was observed in all dogs in control group;while in reconstruction group,the peritoneal membrane was replaced by integrate transitional epithelium,and no obvious stricture was observed.Subepithelial abundant neovascularization was also seen.Conclusions For avulsion of ureteral mucosa exceeding 3 cm,placing stents only will lead to ureteral stricture or atresia,reconstruction using tubularized peritoneal free grafts as mucosa substitutes is an effective method.

Objective To study the effect of genomic HLA-DR compatibility on long-term survival in renal transplantation.Methods A retrospective study was performed on 518 first-cadaver renal transplants by using genotyping technique.Results More than 10%recipients shared HLA-DR matching at DNA level.half of 1 DR mismatches.The recipients with HLA-DR matched transplants showed a significant decrease of acute rejection episodes and a smooth recovery of early renal function as compared with those of DR mismatching kidneys.The 1 to 5 year-person survival rate was increased by 17%to 37.7% (P＜0.01)respectively.Multivariate analysis of 10 variables by Cox regression model revealed that DR mismatching was the most important factors influencing the long-term graft survival.Conclusion Genomic HLA-DR compatibility had a significant impact on long-term survival of first-cadaver kidney transplantation.