Objective To evaluate whether serum cystatin C (SCys C) could be used as an ideal index to assess renal function of renal transplant recipients during posttransplant follow-up.Methods Seventy patients who were followed up for at least 6 months after renal transplantation in our centre were recruited in the study. SCys C and serum creatinine (SCr) were determined during the follow-up period, and glomerular filtration rate (GFR) was measured using an isotope Tc99m DTPA.The correlation between SCys C, SCr and GFR was analyzed. The performance of SCys C and SCr in diagnosing the mild impairment of renal allgraft function (GFR ＜ 1 ml/s) was evaluated using ROC curve. Results Both SCys C and SCr had a linear negative correlation with GFR (r = -0. 82 and -0. 66 respectively, P＜0. 01 ). The sensitivity, specificity and positive predictive values (PPV) of SCys C for diagnosing the mild impairment of renal allgraft function were higher than those of SCr,but the AUC of SCys C did not differ from that of SCr significantly (0. 935 vs. 0. 877, P＞0. 05).Conclusion SCys C could be used an ideal index to evaluate the allograft renal function for its better correlation with actual GFR.

Objective:To construct the cells which can express LIF protein steadily and study the effect of rhLIF and IL-24 on HL-60 cells.Methods:ECV-304 cells were infected by eukaryotic expression plasmid pcDNA3-LIF,then the positive cells were selected by G418.The positive cells were collected and their culture supernatant was stored for further use.Expression of LIF mRNA was detected by RT-PCR.The pAdeasy-1-pAdTrack-CMV-IL-24 was extracted from DH5a.The recombined adenovirus vector was lineared with PacI and transfected into 293 cells.The IL-24 recombined adenovirus(Ad-IL-24) was obtained and used to infect HL-60 cells.At the same time,the culture supernatant containing rhLIF was added into the HL-60 cells which was identified as positive expression of IL-24 by RT-PCR.Synergistic effect of the cytokines on HL-60 cells was tested by LSCM, FCM and immunohistochemistry stain assay.Results:The cells expressing LIF protein steadily were constructed successfully, the high titer of the recombined adenovirus(Ad-IL-24) was obtained. Expression of IL-24 in infected HL-60 cells was identified by RT-PCR.The apoptosis of HL-60 cells induced by rhLIF and IL-24 was proved by LSCM ,FCM and immunohistochemistry stain assay. They had synergistic effect.Conclusion:rhLIF and IL-24 can inhibit growth of HL-60 cells and induce apoptosis of the cells.They have synergistic effect.

Objective To construct the human IL-24 recombined adenovirus(Ad-IL24)and to observe the effect of Ad-IL24 on ex vivo culture of HL-60 cells.Methods pAdTrack-CMV-IL24 was constructed by PCR with pcDNA3.0-IL24 recombined plasmid as a template,enzyme digestion and ligation.The pAdTrack-CMV-IL24 lineared by Pme Ⅰ was co-transformed into BJ5183 with pAdEasy-1.The pAdEasy-1-pTrack-CMV-IL24 recombined adenovirus vector was lineared with Pac Ⅰ and then transfected in to QBI-293A cells.The Ad-IL24 was obtained and used to infect HL-60 ceils,and the effect on HL-60 cells was tested by LSCM,Mrrr,FCM,ELISA and immunohistochemistry staining.Results The pAdEasy-1-pTrack-CMV-IL24 vector was constructed and the Ad-IL24 was successfully obtained.The effect of inhibiting growth of HL-60 cells and inducing cell apoptosis by IL-24 was proved by LSCM,MTT and FCM.IL-24 down-regulated the expression of anti-apoptosis factor Bcl-2 and increased the secretion of IFN-γ,TNF-α of HL-60 cells.Conclusion Ad-IL24 can inhibit the growth of HL-60 cells and induce cell apoptosis through down-regulating expression of anti-apoptosis factor and increasing the secretion of IFN-γ,TNF-α of HL-60 cells.The human Ad-IL24 may provide a basic study for the future target therapy to tumors.

AIM: To develop a new quantitative analysis method for determining diosgenin in Shengmai Freeze-drying Preparation for Injection (Radix Ophiopogonis,etc.)by RP-HPLC with UV detector. METHODS:ODS C_(18) column (5 ?m,4.6 mm?250 mm) was used with mobile phase of acetonitrile-water (88∶12).The mobile phase flow rate was 1.0 mL?min~(-1).The detection wavelength was at 204 nm. RESULTS:The linear range of HPLC was 2.46-7.38 ?g(r=0.999 9).The average recovery was 98.3%,RSD was 0.95%(n=9). CONCLUSION:This method is reliable,accurate and suitable for the determination of diosgenin in Shengmai Freeze-(drying) Preparation for Injection.

Objective To investigate expression and suppressive function of CD39 + regulatory T cells (Treg) in kidney transplant recipients.Method Thirty recipients of first kidney transplants were treated with tacrolimus,mycophenolate mofetil and prednisone.Within 28 days posttransplantation,there were 14 patients subject to acute rejection (AR group),and the rest 16 patients had no episodes of acute rejections (NR group).Twelve healthy volunteers served as healthy controls (HC group).We collected peripheral blood from the three groups and separated PBMC by density gradient centrifugation,and sorted Tresp,CD39-Treg and CD39+ Treg by flow cytometry.We next analyzed the ratio of CD39 + Treg/CD4+ T cells.ELISA was used to determine the suppressive ability of CD39-Treg and CD39+ Treg on secretion of IFN-γ and IL-17 by Tresp.Results The ratio of CD39 + Treg/CD4 + T cells in AR group was significantly reduced as compared with HC group and NR group (P＜0.05).In HC group and NR group,the secretion of IFN-γ and IL-17 by Tresp was suppressed significantly (P＜0.05) by CD39+ Treg.CD39Treg could suppress secretion of IFN-γ but not IL-17 production by Tresp.CD39+ Treg in AR group AR could suppress the secretion of IFN-γ significantly (P＜0.01),but not to IL-17 production.Conclusion CD39+ Treg have important immunoregulation function.The relative amount of CD39+ Treg was reduced and their regulatory function was impaired in patients with acute rejection.

Objective To investigate the efficacy and safety of hypertonie saline enhanced radiofrequency ablation (RFA) in the treatment of liver cancer. Methods The clinical data of 42 patients with primary liver cancer (n = 28) or metastatic liver eancer (n = 14) who had been admitted to First Affiliated Hospital of Nanjing Medical University from September 2001 to December 2007 were collected. Forty-eight lesions were detected with a diameter ranging from 1.2 cm to 7.5 cm. RFA electrode and 20G needle were pricked into the target lesion under the guidance of B ultrasound or computed tomography (CT) through percutaneoas puncture or open approaches. An amount of 5-10 ml hypertonie saline was infused through the needle at regular intervals during RFA. All patients were followed up for 3-79 months. Contrast-enhanced ultrasound and CT scanning were performed postoperatively to determine the efficacy of RFA. The levels of alpha-fetoprotein (AFP) before and after treatment were compared using t test, and the survival of the patients were analyzed using a Kaplan-Meier survival curve. Results The AFP expression changed to negative in 14 out of the 18 AFP-positive patients, with statistical difference (t =7.703, P <0.05). The complete necrosis rate of tumors was 94% (45/48), and the necrosis rate of tumors with diameter of ≤4.0 cm reached 100% (35/35). The incidence of complication was 5% (2/42). No perioperative mortality occurred. The 1-, 2-, 3-year survival rates were 91%, 85% and 70%, respectively. Conclusions Hypertonic saline enhanced RFA in the treatment of liver cancer was proved to be safe and effective.

Objective To investigate a possible association of donor-recipient compatibility for ABO blood group alleles with acute rejection (AR) in renal transplantation. Methods A study comprising 87 pairs of donor and recipient was performed. The ABO genotype A1, A2, O1, O2, and B alleles of renal transplanted recipients and their respective donors were assessed by PCR amplification with sequence-specific primers (PCR-SSP). Accordingly, recipients were divided into donor-recipient ABO genotype matched and mismatched groups. Results The PCR-SSP based types of all cases showed total concordance with their serologically assigned ABO groups. Fifty pairs (57. 5%) were matched for ABO genotype among the 87 pairs of donor and recipient while 37 (42. 5%) were mismatched, including 1 allele mismatch in 31 pairs (83.8%), 2 alleles mismatches in 6 pairs (16. 2%).The incidence of AR was 12.0% (6 cases) and 29. 7% (11 cases) for ABO genotype matched and mismatched transplant patients, respectively ( P ＜ 0.05). After high dose methylprednisolone (MP)treatment, all cases exepienced reversion of AR except a A2O1 recipient receiving kidney from a A1O1enced 4 AR episodes within 3-10 months, and the period of AR was gradually shortened. After high dose MP was administered empirically, even though short-term improvement of renal function was observed, the serum creatinine continued to increase progressively with decreased efficacy of high dose MP. One year after operation the serum creatinine rose to 441 μmol/L. Conclusions Simultaneous definition of the ABO genotype and HLA is highly feasible. The A2 patient is suitable for receiving kidneys from blood group O donors. DNA mismatch for ABO genotype of renal transplant recipients and their respective donors is an independent risk factor for AR. Genotyping of ABO blood group is conducive to prevent AR.

Objective To summarize the clinical experience of combined liver and kidney transplantation (CLKT). Methods CLKT was performed on 22 patients. The orthotopic liver transplantation (LT) was preceded with the classic fashion in 10 patients and piggyback fashion in 12 patients. The renal allograft was implanted to the iliac fossa routinely. After operation, the patients received an induction therapy with anti-CD25 monoclonal antibody or antithymocyte globulin ( ATG) and a maintenance therapy with tacrolimus (Tac), mycophenolate mofetil and prednisone. Results The CLKT was successfully performed on all 22 patients, and the graft function was restored well postoperation. During the perioperative period, an acute rejection episode of liver occurred in one patient and acute renal allograft rejection episode in 2 patients. The Tac toxicity occurred in one patient. The hemorrhage of digestive tract occurred in one recipient and the hemorrhage of peritoneal cavity in one patient. The pleural effusion occurred in 6 recipients. The pneumonia occurred in 2 cases and the peritoneal infection in one patient During a follow-up period of 6 months to 7 years 11 months, three patients died because of cytomegalovirus pneumonia in 2 patients and acute myocardial infarction in, one patient, The 1-, 3-, 5-year survival rate of recipients was 86,4 %, 81.3 %, 72.7 % respectively. Conclusion The CLKT is an effective method for treatment of patients with end-stage liver djsease and chronic renal failure.

Objective To explore the relationship of serum anti-MICA antibody and development of chronic rejection (CR) after renal transplantation. Methods The enrolled 105 patients included 43 cases of CR, and 62 cases of functioning renal allograft as controls. Data including PRA level before transplantation, HLA mismatch, cold ischemic time, SCr at discharge, immunosuppressive regimen,and months after transplantation were analyzed. Blood samples were collected immediately after grouping for anti-MICA antibodies, SCr determination. Acute rejection episodes and renal allograft function which was evaluated by △SCr/M [(SCr at present - SCr at discharge) /months after transplantation) were compared between anti-MICA-antibody positive patients and anti-MICA-antibody negative patients. Results There was no significant difference in gender, age, HLA mismatch, cold ischemic time, immunosuppressive regimen, SCr at discharge, months after transplantation between CR and control groups (P＞0.05). Serum creatinine level and number of antiMICA-antibody positive patients in CR group were significantly increased as compared with those in control group (P＜0.01 ). Acute rejection episodes during the first 3 months after transplantation in anti-MICA-antibody positive patients were significantly more than those in anti-MICA-antibody negative patients (P＜0.05),and the △SCr/M in the former was higher than that in the latter (8.3 +3.6 vs 2.4 ± 2.6, P＜0.05). Conclusion Humoral immunoreaction mediated by MICA partly participates the development of CR after renal transplantation. MICA antibody is a risk factor affecting long-term allograft function.

OBJECTIVE To explore the relationship between urinary tract infection(UTI) and urological surgical site infections(SSI).METHODS The surgical site infections of 115 patients underwent urological operations were observed.Patients were screened for the presence of UTI before the operation and subcutaneous swabs for culture were collected at the end of the operation by brushing with a sterile cotton-swab just before skin closure.The resistance and similarity of the isolates were analyzed by antimicrobial susceptibility test and pulsed field gel electrophoresis(PFGE).RESULTS Surgical site infection occurred in 14 of 115 patients.The isolates from infected surgical sites showed higher resistance to antimicrobial agents than those from non-infected surgical sites(P