Objective To investigate the experiences of combined liver-kidney transplantation (CLKTx).Methods Six patients underwent CLKTx in our center. The primary diseases included chronic glomerulonephritis and post-hepatitis B cirrhosis in 4 patients, hepatitis B virus associated nephritis and primary hepatocellular carcinoma in 1 patient, and polycystic kidney and polycystic liver in 1 patient. Cyclosporine A (or tarcrolimus), mycophenolate mofetil and methylprednisolone were applied to prevent rejection. Four cases of post-hepatitis B cirrhosis received lamivudine. And hepatitis B immunogloblin was given to 4 patients for a short term.Results All 6 liver grafts had good primary function. Five renal grafts had good primary function within one week post-transplantation. One patient with delayed kidney graft function needed supportive hemodialysis. The serum creatinine was declined to normal level 52 days post-operation. Pleural effusion occurred in all 6 patients among which 2 patients needed surgical drainage. Two patients had to be treated for bacterial pneumonia and pneumocystis carinii pneumonia respectively. Three patients needed lipid-lowering therapy at early time post-operation. At the last follow-up, all 6 patients were alive with normal renal graft function and liver graft function. The panel reactive antibody (PRA) of one patient was 23 % before transplantation, and remained at about 8 % post-transplantation. The serum HBsAg and HBV DNA of all 4 post- hepatitis B cirrhosis patients became negative post-transplantation.Conclusion CLTx is a safe procedure for combined hepatic and renal end-stage disease with excellent short-term results.

Objective To investigate whether the mortality and death causes of renal transplant recipients have changed from 1985 to 2002 in our center.Methods 145 cases of renal transplant recipients who died during 1985 to 2002 were divided into 3 groups: 1985 to 1990,1991 to 1996,1997 to 2002. The death causes and the mortality on the 1st ,2nd,3rd,4th and 5th year post transplant of each group were reviewed.Results During the three periods,the 1st,2nd,and 3rd year mortality was decreased. Infection as a cause of death fell from 31.8 % to 29.2 % and 26.7 % . Whereas death from liver disease,cancer and cardiovasvular diseases was increased.Conclusions The mortality is decreased. Infection,cardiovascular diseases,liver diseases and cancer were the main causes of death after transplantation from 1985 to 2002. It is important to prevent these diseases and treat them effectively in order to improve the recipients' survival rate.

Objective To revise the Chinese version of the National Institutes of Health-Chronic Prostatitis Symptom Index (CHN-NIH-CPSD), and evaluate its feasibility, reliability, validity and responsiveness. Methods The NIH-CPSI was translated into Chinese according to a standard methodology including forward-backward-forward technique. The CHN-NIH-CPSI was pre-tested in consecutive samples of 162 native-speaking Chinese chronic prostatitis(CP)patients. Ninety-five of 162 filled the index again on the same day and after 4-week therapy. Ninety-seven healthy men were included as evaluated. Results The recovery of the questionnaires was 100% and all the patients filled the index completely. The mean time to complete the questionnaire for the patient group was 5.2±2.4 (range 2 - 12) min. The split-half reliability was 0.82. For the overall index and each subscale, the test-retest reliability was 0.98, 0. 98, 0. 98, 0. 97, respectively(P＜0.01);and the Cronbach's α coefficient was 0. 61,0. 71, 0. 59, 0. 75, respectively. The confirmatory factor analysis showed good construct validity with a goodness of fit index of 0. 85 and a x2 of 124.67(P＜0. 01). Of all 162 patients, the scores of the overall index and each subscale were 23. 33±5.91. 8. 80±4.26, 5.30±2.82, 9. 23±1.90, respectively;and those of healthy controls were 1. 95±1.97, 0. 37±1.03, 0. 15±0.58, 1.42± 1.20,respectively. Of the 95 patients, the original scores were 23. 53±5.60, 9.21 ±4.04, 5.10±2.75,9.21 ±2.05, comparing with 19.47±6.36, 7.79±3.95, 3. 58±1.88, 8.11±2.50, the 4 weeks later scores. The group t-test and paired t-test showed good responsiveness. Conclusions The CHN-NIH-CPSI has high feasibility, reliability, validity and responsiveness for testing the patients with CP. It is suitable for Chinese-speaking patients and helpful for cross-cultural comparisons of men with CP in clinical and research settings.

Objective To investigate the effect of anti-apoptotic protein HSP-90 in human multiple myeloma cell line U266 cells after bertezomib interaction. Methods The HSP-90 mRNA expression in the U266 cells was tested by reverse transcription polymerase chain reaction (RT-PCR) after 4 hours of treatment of bertezomib by different concentration. Results With the of increased concentration of bortezomib, the expression level of HSP-90 αmRNA was also increased in U266 cells. Respectively, quantitative results of HSP-90α are 0.343±0.017, 0.505±0.039, 0.640±0.029, 0.760±0.059, 0.963±0.054 from the low to high concentration of bertezomib groups. And there are statistical difference between each group(P <0.05). However, the HSP-90β quantitative results in 0 nmol/L concentration of bertezomib (0.61±0.022) have statistical difference between 50, 150, 200 nmol/L groups(P <0.05). HSP-90β quantitative results in 50(0.765±0.050)and 100 nmol/L(0.645±0.052) nmol/L groups are different(P <0.05). Compared with 100 nmol/L concentration of bortezomib group, statistical difference also exists in 150 (0.770±0.059) and 200 nmol/L (0.790±0.027)groups (P <0.05). Although there is no obvious increase in the mRNA expression of HSP-90β from the chart, statistical difference existed in the whole data (P <0.05). Conclusion Bortezomib can increase the level expression of HSP-90 mRNA, and especially increase the level expression of HSP-90α mRNA.

Objective To evaluate the effects of tadalafil on chronic allograft vasculopathy. Method The abdominal aorta transplantation was performed on Male Lewis or Brown-Norwai rats as donors and male Male Lewis rats as recipients. The recipients were divided into 3 groups: Group A (Lewis-Lewis)with no treatment ;Group B (BN-Lewis) with no treatment; Group C (BN-Lewis) received tadalafil treatment (0.5 mg/kg per day). The rats were sacrificed at 8 weeks post treatment. The grafted aortas were used for histology and Western blot assay. Plasma cGMP level was detected by ELISA assay. Results The aortas intimal in group C was significantly thinner than that in group B. PKG-Ⅰprotein expression in group C was significantly higher than that in group B. Expression of RhoA in group C was lower than that in group B. Conclusion Tadalafil has positive effect on chronic allograft vasculopathy.

0.05).The optimal combination was PCR-CTPP for MDR1 C3435T and PCR-SSP for G2677T/A.Conclusions PCR-CTPP and PCR-SSP are simple,accurate,rapid and economical methods for detection of SNP of MDR1 C3435T and G2677T/A,and can be applied in clinical research.

Objective To study the mechanism of arachnoidal fibrosis after subarachnoid hemor- rhage. Methods Rats were divided into control group, experiment group and treatment group. Radioim- munoassay (RIA) was employed to detect the levels of hyaluronic acid, laminin,type Ⅲ precollagen and type Ⅳ collagen in the arachnoid membrane. In the meantime, arachnoid cell's morphology and collagen distribution in the subarachnoid space were investigated by electron microscope. Results Results of RIA detection showed increase of Type Ⅲ precollagen level (peak at the second week), obvious higher levels of LN and HA but insignificant change of type IV collagen after subarachnoid hemorrhage. However, dexam- ethasone treatment decreased type Ⅲ precollagen level. Electron microscope found that arachnoid cells pres- ented accentruated bioactivity after subarachnoid hemorrhage, with significant increase of arachnoidal colla- gen fibers from one week after suharachnoid hemorrhage, continuing for 3 weeks. Dexamethasone treatment resulted in low density of mitochondria and sparsed arachnoidal collagen fibers. Conclusions Extracellu- lar matrix (ECM) increases in arachnoid membrane after subarachnoid hemorrhage and participates in a- rachnoid fibrosis. Dexamethasoue can relieve arachnoidal fibrosis after subarachnoid hemorrhage, as pro- rides fresh way for prevention and treatment of post hemorrhagic hydrocephalus.

Objective To explore the feasibility and effect of combined piggyback orthotopic liver and kidney transplantation in patient with renal graft function failure complicated with cirrhosis. Methods One patient with renal graft function failure complicated with cirrhosis was resected of renal graft. Daily dose of 50?mg CTX was given 5 days after operation and continued for 3.5 months. After two courses of plasma exchange, PRA was reduced from 66?% to 23?% . Combined kidney and liver transplantation was performed simultaneously using piggyback orthotopic liver transplantation technique,and the kidney graft was placed retroperitoneally in left iliac fossa. PRA was monitored every 30 min after liver reperfusion. Postoperative immunosuppressive therapy consisted of FK506, MMF and steroids. Results The kidney and liver grafts functioned normally after transplantation. PRA was reduced from 23?% to 5?% and maintained at 8?% . HBsAg and HCV were returned to negative. Kidney and liver grafts functioned well during a follow up of 3 months. Conclusion Combined liver and kidney transplantation is an effective rescue for loss of kidney graft complicated with cirrhosis, and liver graft can provide protection towards the kidney graft from the same donor.

Objective To explore the molecular mechanism of renal function defects after urinary obstruction and investigate the effect of sirolimus on the expression of γ-ENaC,Na + K + ATPase and AQP2,and its mechanism of renal Water-Electrolyte imbalance following bilateral ureteral obstruction (BUO) in rat kidneys.Methods Forty-eight rats were randomly divided into a sham operation group ( sham group),a BUO group,and a sirolimus treatment after BUO group.Bilateral ureters were exposed and occluded with ligature in the BUO and sirolimus treatment groups.Twenty-four hours later,the obstructed ureters were decompressed by removal of the ligature.The sham animal group underwent identical surgical procedures,but the ureter was simply dissected without removal of the ligature.The sirolimus treatment groups was given sirolimus intragastricly 0.4 ml per day (2 mg/kg · d) from the day before surgery until the rats were scari fled.The sham and BUO groups were given the same volume of intragastric saline.The urine and blood were collected at 4 d,7 d after surgery,and the functional data were observed.The expression of γ-ENaC,Na+K + ATPase and AQP2 were examined by immnohistochemistry and immunoblotting.Results On day four and seven post ureteral obstruction release,urine volume in the BUO group were (85.31 ± 13.15,66.39 ±10.56 ml),significantly higher than that of sham operation (35.36 ± 7.74,33.90 ± 8.03 ml) and sirolimus treatment groups (69.81 ± 10.70 ml,48.57 ± 9.01 ml) (P ＜ 0.05 ).Urine sodium concentrations in the BUO group were (42.17 ± 7.35 mmol/L,43.63 ± 18.39 mmoL/L),significantly lower than that of sham operation ( 170.56 ± 18.39 mmoL/L,172.52 ± 7.35 mmol/L) and sirolimus treatment groups (76.18 ± 13.20 mmol/L,134.28 ± 13.20 mmol/L),P ＜ 0.05.Immunoblotting assay showed that,on day four and seven post rats ureteral obstructions were released,integral optical density of γ-ENaC (2.09 ±0.32,2.27 ±0.35),Na+ K+ATP enzyme (2.41 ±0.48,2.67 ±0.43) and AQP2 (2.17 ±0.45,2.63 ±0.28) in the sirolimus treatment group were significantly higher than those of BUO group ( 1.28 ± 0.21,1.45 ±0.17) (1.99 ±0.28,2.18±0.24) (0.93 ±0.22,1.31 ±0.16),but still lower than the sham group (2.58±0.51,2.60±0.56) (2.89±0.53,2.97 ±0.66) (3.05 ±0.63,3.10±0.67).There were significant differences among all the three groups ( P ＜ 0.05 ).Conclusions The downregulation of γ-ENaC,Na + K + ATPase and AQP2 expression after BUO may contribute to the impaired renal tubular sodium reabsorption,decreased urinary concentration,and postobstructive polyuria.Sirolimus treatment significantly prevents impairment in renal function and also prevents downregulation of y-ENaC,Na + K+ ATPase and AQP2during BUO,demonstrating a marked renoprotective effect of sirolimus treatment in conditions with urinary tract obstruction.

Objective To establish a universal stem loop primer (USLP) based real-time PCR method to scan mature miR profile and quantify it's expression.Methods The common universal stem-loop primer pairs were re-designed; 8 random nucleotides were introduced at 3 ' end for reverse transcription of the mature miR,establishing a miR scanning and quantifying system based on SYBR Green Ⅰ PCR (improved USLP method).10-fold gradient diluted standard miRNA-155 cDNA ( 1 ～ 109 copies/μ1) were utilized to evaluate the sensitivity of this method.The specificity was verified by melting curve assay; the precision was assessed by intra-assay coefficient of variation (ICV) of threshold cycle (Ct value) through 20 repeated detections of the standard miR-155 cDNA (2 × 105,2 × 106,2 × 107 copies/μl) ; cost of the primers and time were evaluated,compared with that of the conventional USLP method.Peripheral blood samples were cultured with phytohaemagglutinin (PHA) for0 h,16 h,24 h,48 h and 72 h,and 87 candidate miR that may be associated with human immunity from PubMed data were scanned and quantified from the cultured T cells.Results The sensitivity of the improved USLP method was 103 copies/μl of standard miR-155 cDNA.Melting curve assay showed a single melting peak at 80 ℃,suggesting the excellent PCR specificity of miR-155.Precision of our method quantifying miR-155 was acceptable (ICV ＜ 2.5％ ).Compared with the traditional stem loop primers,our method saved 75％ cost of primers ( 1 917 bp vs 7 851 bp) and 60％ test time of reverse transcription (85 min vs 205 min).By our method,85 of the 87 miR expression in T cells had no significant difference after the PHA stimulation; the expression of miR-150 (72 h) decreased by 10 times and that of miR-155 (48 h) increased 8 times after culture with PHA (Z =-2.032,P =0.042;Z =- 2.023,P =0.043,respectively ).Conclusions The improved USLP method is fast,precise,sensitive,and cost-effective.It could be used for miR profile scanning and quantifying in T cells.