Objective To assess the immune tolerance of allogeneic T cells induced by dendritic cells genetically engineered to express I?B? mutant. Methods DCs were prepared from WF rat bone marrow cells and modified by I?B?M gene wit h adenovirus vector, and then the expression of I?B? and I?B?M was detected by Western-blot. The cell-surface expression of costimulatory molecules (CD80, CD86 and CD40) was detected by flow cytometry, and the production of IL-12 in DCs culture supermatant was determined by ELISA. The ability to stimulate the pr oliferation of Lewis rat T cells was analyzed by mixed leukocyte reactions (MLR) . The antigen-specific T cell hyporesponsiveness was tested by secondary MLR. Results I?B?M suppressed the cell-surface expression of costimulatory molecules and i nhibited the production of IL-12. I?B?M-DC showed reduced ability to stimula te T cells proliferation, and potential to induce antigen-specific T cell hypo responsiveness. Conclusion Immune tolerance of T cells can be induced by dendritic cells genetically engine ered to express I?B? mutant.

Objective To evaluate the effect of endothelin A receptor antisense oligodeoxynucleotides (ETAR-ASODN)on the growth of human prostatic stromal cells. Methods Primary cultured prostatic stromal cells were derived from patients with benign prostatic hyperplasia (BPH).The cells of passages 4～6 were routinely used for this study after identification.ETAR-ASODN at the concentrations of 5,10 and 15 ?mol/L were added into the culture cells with lipofectin, and cell proliferation was measured by MTT assay. The expression of ETA receptor was tested by 125 I-ET-1 radioligand banding assay. Results MTT assays showed a significant decrease in cell proliferation in stromal cells after 5,10 and 15?mol/L ETAR-ASODN were added with the A 540 values being 0.304?0.082,0.296?0.008 and 0.194?0.061,respectively.The proliferative activity was significantly decreased compared with control group ( P

Objective To investigate the expression of endothelin 1 (ET 1) and ETA/B receptors mRNA in prostatic tissues and its clinical significance in patients with benign prostatic hyperplasia (BPH). Methods Immunohistochemical staining and RT PCR were used to detect the expression level of ET 1,ETAR mRNA and ETBR mRNA,respectively.The detection results were analyzed in correlation with the clinical parameters of BPH patients. Results The expression level of ET 1 and ETA/BR mRNA in BPH tissue(integral optical densities,0.94?0.08,0.64?0.08,0.97?0.08,respectively)was higher than that in normal prostatic tissue(0.57?0.06,0.37?0.05,0.51?0.04,respectively).In BPH group,the expressed quantities of ET 1 and ETAR mRNA showed positive correlation with IPSS,prostatic volume,prostatic urethral length,prostatic urethral pressure and maximum urethral pressure of BPH patients,but it showed negative correlation with maximum flow rate and average flow rate of them. Conclusions Increased expression of ET 1 and ETAR might play a role in bladder outflow obstruction(BOO) of the patients with BPH.

Objective To observe the effects of Triptolide on the expression of toll-like receptor 4 (TLR4) in renal ischemia/reperfusion (I/R) injury in rats. Methods A renal I/R model was established. Rats were randomly separated into the following experimental groups. Group 1, shamoperated control (n = 15) : rats were subjected to surgical manipulation, without the induction of renal ischemia. Group 2, I/R (n = 18): rats were subjected to left renal ischemia for 45 min followed by reperfusion. Group 3, TRI + I/R (n = 18): Before the I/R procedure (as in group 2), rats were intraperitoneally injected with TRI (0.4 mg/kg), once every day, three times. Rats were killed at the 1st, 3rd, and 5th day after I/R injury. The parameters of renal function were determined by autobiochemical analyzer. The expression of TLR4 was detected by RT-PCR and Western blotting. Results As comparedwith the sham-operated control group, serum BUN and Cr levels were significantly increased in the rats undergoing I/R procedure at the 1st, 3rd, and 5th day (P＜0. 01). After the treatment with TRI, the levels of BUN and Cr and the expression of TLR4 in the renal tissues were significantly decreased (P＜0. 05). Conclusion TRI could relieve renal I/R injury in rats by inhibiting the TLR4 expression.

ObjectiveTo investigate the protective effect of ozone oxidative preconditioning (OzoneOP) on apoptosis induced by acute renal ischemia/reperfusion (I/R) injury in rats.Methods The right kidneys of rats in control group were excised.A rat I/R model was established in I/R group.In OzoneOP group,the renal OzoneOP was induced by rectal insufflation of 5.0～5.5 ml oxygen plus ozone (ozone 50 mg/L,1 mg· kg- 1 · d- 1,once every day).The parameters (blood urea nitrogen and creatine )of renal function were determined by auto-biochemical analyzer. The concentrations of malondialdehyde (MDA) and superoxide dismutase (SOD) were measured by using chromometry.Cytochrome c (CytC) was examined by using immunohistochemistry and Western blotting.The mRNA expression levels of TNF-α,IL-1β and IL-6 in the renal tissue were detected by using RT-PCR.ResultsCompared with the control group,the blood urea nitrogen,creatine,MDA and the release of CytC were increased significantly in I/R group.After OzoneOP,the levels of blood urea nitrogen,creatine,MDA,CytC and the mRNA expression levels of TNF-α,IL-1-β and IL-6were significantly decreased and SOD levels were significantly increased.ConclusionOzoneOP could relieve renal I/R injury of rats by improving antioxidation capability,cutting down cytokines contents and inhibiting the release of CytC from mitochondria.

Objective To investigate the influence of m4-1BBL on the anti-tumor effects induced by truncated human prostate specific membrane antigen (tPSMA) gene in mice. Methods A eukaryotic expression plasmid encoding tPSMA and m4-1BBL (pDC316-tPSMA-IRES-m4-1BBL), pDC316-tPSMA and pDC316 were constructed. C57BL/6 mice were vaccinated in the quadriceps femoris, respectively. The CTL activity of spleen cells from the immunized mice against prostate cancer RM-1-tPSMA was detected by CCK-8 kit in vitro. The tumor growth was then observed. Results The target cell specific cytotoxicity rate induced by pDC316-tPSMA-IRES-m4-1BBL was 42.6%, compared to 24.8% in the pDC316-tPSMA group and 10.8% in the pDC316 group. The difference was significant (P<0.05). The volume of tumor in the pDC316 group was 2657.4mm3 7 d after vaccination, compared to 1334.5 mm3 in the pDC316-tPSMA group, 9 d after vaccination. In the pDC316-tPSMA-IRES-m4-1BBL group, the tumor volume was 445.8 mm3, 12d after vaccination. The difference was significant (P<0.05). Conclusion Gene vaccines co-expressing tPSMA gene and m4-1BBL gene could significantly enhance anti-prostate cancer effects in mice.

Objective To evaluate the predictive value of urinary neutrophil gelatinase-associat-ed lipocalin (NGAL) and interleukin-18 (IL-18) for delayed graft function (DGF) in kidney transplan-tation. Methods Serial urine samples collected at 0, 12 and 24 h after operation from 86 kidney transplantation patients were analyzed by enzyme-linked immunosorbent assay for NGAL, IL-18 and RBP. Results Fifteen patients developed DGF. At 12 h after transplantation, the level of urine NGAL elevated significantly (1712.75±474.6 vs. 863.1±199.8 without DGF, P<0. 001). The in-creases of urine IL-18 (29. 2±4.1 vs. 28.7±4.2 without DGF, P>0. 05) was not significant. At 24 h, both urine NGAL(2905.0±1108.1 vs. 911.8±221.0 without DGF,P<0. 001) and IL-18(211.3± 34.0 vs. 86.9±22.8 without DGF, P<0. 001) increased significantly, whereas the changes of urine RBP and serum creatinine (SCr) were not significant. ROC analysis showed that the area under the curve of NGAL and IL-18 were 0. 90 and 0.76 respectively, the cut-off values were 996.5 ng/mg and 148.5 ng/mg, the diagnostic sensitivities in DGF were 90.2% and 76.3%, specificities were 82.6% and 66.4% respectively. Conclusions Both urine NGAL and IL-18 could potentially be early predic-tive marker of DGF. The level of NGAL elevated earlier than IL-18, which may be more effective in predicting DGF.

PURPOSE: The purpose of this study is to construct a recombinant adenovirus vector carrying mouse 4-1BBL and observe its effects in dendritic cells. MATERIALS AND METHODS: Mouse 4-1BBL cDNA was taken from the plasmid pcDNA3-m4-1BBL and subcloned into adenovirus shuttle plasmid pAdTrack-CMV, and then transformed into competent BJ5183 with plasmid pAdEasy-1. After recombination in E. coli, Ad-4-1BBL was packaged and amplified in HEK 293 cells. The expression of 4-1BBL in Ad-4-1BBL-transfected mouse prostate cancer cell line RM-1 was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. After the co-culture of dendritic cells (DCs) with Ad-4-1BBL-transfected RM-1 cells, interleukin (IL)-6 and IL-12 production were assessed by enzyme-linked immunosorbent assay (ELISA) and co-stimulatary moleculs (CD80 and CD86) on DCs were analyzed by flow cytometry. RESULTS: The levels of IL-6 (3,960 pg/mL) and IL-12 (249 pg/mL) production in Ad-m4-1BBL-pulsed DCs were more than those in none-pulsed DCs. The differences were statistically significant (p < 0.05). The expression of co-stimulatary molecules (CD80 and CD86) was up-regulated in Ad-m4-1BBL-pulsed DCs. CONCLUSION: The results indicated the recombinant mouse 4-1BBL can effectively activate DCs.