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.

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 compare the efficacy and safety of Tacrolimus(FK506)and Neoral CsA in conjunction with MMF(2.0g/d)and steroid in preventing renal allograft rejection.Methods 98 cases of renal transplant recipients were randomly divided into two groups:FK506 group(n=40),receiving tacrollimus,MMF and prednison(Pred);CsA group(n=58),receiving CsA,MMFand Pred.Results The mean follow-up time in both two groups Was 12.5 months.Acute transplanted renal rejection occurred in 2 cases in FK506 group and 9 cases in CsA group respectively.The one-year person/kidney survival rate was 100%/100%in FK506 group and 100%/94.8%in CsA group respectively.The dosage of Pred in FK506 group was lower than in CsA group.12 cases in FK506 group had stopped using Pred.Hypergly cermia occurred in 7 cases in FK-506 group.Polytricosis,gingival hyperplasia and liver function disorder dominantly occurred in CsA group.Infection Was found in 9 cases of FK506 group and 11 cases of CsA group respectively.Conclusion FK506 combined with MMF could decrease the occurrence of acute trans planted renal rejection and the dosage of Pred.The good adjustment of the dose of FK506 iS helpful for re ducing the side effects and preventing rejection.

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 investigate the effect of nuclear factor ?B(NF-?B)decoy on the chemokine expression in bladder cancer cell line. Methods Human bladder cancer cell line EJ,NF-?B decoy ODN were used as a NF-?B inhibitor(scrambled NF-?B decoy was used as control).NF-?B DNA binding activity was detected by electrophoretic mobility shift assay (EMSA);and p65 subunit of NF-?B was detected by RT-PCR and Western blot.Chemokines including IL-8,MCP-1,RANTES were detected by RT-PCR. Results EMSA showed that NF-?B decoy inhibited NF-?B activation induced by TNF-?.RT-PCR or Western blot test suggested that p65,IL-8,MCP-1 and RANTES were upregulated by TNF-? and downregulated by NF-?B decoy.However,mutated decoy ODN had no effect on them. Conclusions Chemokines can be detected in bladder cancer.They are activated by TNF-? and inhibited by NF-?B decoy.

Objective To observe the dynamic change of cytokines and effect of different stimulating combinations on dominant regulation, and describe the microenviroment character of local immune response. Methods PBMCs were separated from peripheral blood of healthy donors with density gradient centrifugation. The expression levels of cytokine genes under different stimulating combinations, including IL-2, IFN-?, IL-12, IL-4, IL-10 and TGF-?1, were detected by ELISA and RT-PCR. Results ELISA results indicated that secretion level of IL-2, IL-4, IL-12 and IFN-? were inhibited by IL-10. There were significant differences in IL-4 and IL-12 between control group and stimulating groups under the stimulation by anti-CD3 monoclonal antibody (McAb) or anti-CD3 McAb + anti-CD28 McAb combination. Significant differences in IL-2 only appeared in anti-CD3 antibody stimulating group. The concentrations of IFN-? were decreased moderately. At the mean time, IL-10 significantly promoted the secretion of TGF-?1 under the anti-CD3 McAb stimulation. There were similar results on the genes level for the studied cytokines. Conclusion The induction and maintenance of dominant regulation were dependent mainly on IL-10, which inhibits the overexpression of IL-4 and IL-12. It is the vital step for dominant regulation to avoid the mono-polarization development at the beginning of immune response.

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 establish and validate andro ge n-independent human prostate cancer LNCaP cell model. Methods Androgen-dependent LNCaP parental C-33 cells were maintained in a regul ar cell-culture medium,that is,phenol red-positive RPMI 1640 medium supplement ed with 10% fetal bovine serum,1% glutamine and 0.5% gentamicin.Upon continuousl y passaging,androgen-dependence of these LNCaP cells decreased gradually,thus,t he androgen-independent LNCaP cell model which derived from androgen-dependent cells was established. Results Upon continuous passage, the biological behavior of androgen-dependent parental LNCaP C-33 cells (pass age number less than 33) was altered.LNCaP C-81 cells (passage number higher th an 80) clearly exhibited more aggressive growth and lower androgen-dependence t han C-33 and C-51 cells (passage number between 34 and 81) in vitro and in viv o. Conclusions The LNCaP cell model closely recapitulate s the transition of human prostate cancer from androgen-dependent to hormone-r efractory state under the androgen nondeprived condition. This cell model may pr ovide the opportunity to understand the molecular mechanisms involved in the and rogen-independent growth of cancer cells during prostate cancer progression.

ObjectiveTo investigate the effect of NF-?B on the differentiation and maturation of mu rine bone marrow-derived dendritic cells (DC) in vitro.MethodsThe activity of NF-?B in dendritic cells was blocked by oligodeoxyribonucleot ide Decoys (ODN Decoys) containing two special binding sites for NF-?B. Morpho logical changes of DS were observed. The expression of assistant molecules on th e cell surface was detected by using flow cytometer, and the stimulating activit y of allogeneic T lymphocyte proliferative response was determined in culture mu rine DC. The effect of NF-?B on the maturation and immunobiological activity o f murine DC was studied.ResultsNF-?B ODN Decoys were efficiently incorporated by DC, markedly suppressed the maturation of DC, the expression of assistant costimulatory molecules (CD80, CD8 6 and CD40) on the surface of DC and the secretion of IL-12, blocked the develo pment of DC and this blocked function was not reversed by lipopolysaccharide (LP S). In mixed lymphocyte reactions, DC treated with NF-?B ODN Decoys could indu ce allogeneic T cells hyphoresponsiveness, and this was associated with the inhi bition of Th1-type cytokines (IL-2 and IFN-?) production.ConclusionsNF-? B is a key gene to the development and maturation of DC. Specific interference w ith NF-?B in DC using ODN Decoys approaches could offer a novel strategy for i nducing and generating tolerogeneic immature DC and provide a promising means to induce transplant immune tolerance.

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