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 study the extraction technology of ursolic acid from sambucus chinensis Lindl. Methods: The method of ethanol extraction and agglutination separation was adopted for extracting ursolic acid. Results: The extraction rate was 90%, its purification was 98%, the product was recognized to be ursolic acid by physicochemical contents and spectral identification. Conclusion: This method is advanced, practical, reasonable and feasible. It can be applied in industrial production.

Objective To systematically assess the diagnostic value of gene detection for spontaneous bacterial peritonitis (SBP) .Methods A literature search was performed in the database of PubMed ,Web of Science ,Cochrane Library and China National Knowledge Internet (CNKI) from databases establishing to March 2015 .Relevant studies on diagnostic value of gene detection for SBP were retrieved .Quality assessment of diagnostic accuracy studies (QUADAS ) was applied for the included studies .Meta-analysis was conducted using bivariate random effects model .Summary receiver operator characteristic curves (SROC) was conducted to calculate area under curve (AUC) and was compared using Z test .Results Five studies with 423 specimen involved were included in the meta-analysis .The pooled sensitivity ,specificity ,diagnostic odds ratio (DOR) ,positive likelihood ratio and negative likelihood ratio of gene detection for the diagnosis of SBP were 0 .56 (95% CI:0 .49 -0 .62) ,0 .88 (95% CI:0 .83 -0 .92 ) ,9 .94 (95% C I:1 .76-56 .27 ) ,4 .35 (95% C I:1 .05 -18 .10 ) and 0 .47 (95% C I:0 .25 -0 .88 ) , respectively .The pooled sensitivity was significantly higher than that of bacterial culture (0 .25[95% CI:0 .19-0 .31]) .The AUC of SROC of gene detection was 0 .810 9 ,which was significantly higher than that of bacterial culture (AUC=0 .659 8 ,Z=3 .14 ,P<0 .01) .Subgroup analysis was conducted in patients with polymorphonuclear neutrophils (PMN)≥250 × 106/L in ascites .All the diagnostic indices of gene detection were inferior to those of bacterial culture for SBP ,except for the sensitivity of gene detection for SBP (0 .64[95% CI:0 .53 -0 .74] vs 0 .39[95% CI:0 .29 -0 .51]) .The diagnostic value of quantitative polymerase chain reaction (qPCR) detection for SBP was inferior to that of bacterial culture in all the aspects except for the sensitivity (0 .54 [95% CI:0 .47 -0 .61 ] vs 0 .25 [95% CI:0 .19 -0 .31 ]) . Conclusions Gene detection shows higher sensitivity than bacterial culture .The diagnostic value of gene detection is influenced by diagnostic standards .qPCR also shows high sensitivity for SBP diagnosis ,while the diagnostic value was inferior to bacterial culture .More researches with high quality are required to validate the results of this study .

Objective:Expression and purification of the α-HL of Staphylococcus aureus as antigen for making full human anti-α-HL antibody later ,providing of new treatment for Staphylococcus aureus infection.Methods:The total RNA of Staphylococcus aureus was extracted and the cDNA of α-HL was amplified by RT-PCR.The DNA of α-HL and pCold-TF plasmid was digested and ligated by T4 ligase and then transformed into E.coli TOPO 10.The recombinant plasmid α-HL/pCold-TF which verified by sequencing was trans-formed into E.coli BL21 for expression.The expression products was identified by SDS-PAGE and Western blot.Results: The size of amplified cDNA of α-HL was about 900 bp and the expressed soluble fusion protein of α-HL was about 90 kD(including the molecular chaperone in the vector ) after inducing expression for 24 h at 15℃.The Western blot results showed that the expressed protein was the fusion protein of α-HL.The purified α-HL was injected into BABL/c mice for making antiserum.The results showed that the antiserum had good binding activity with Staphylococcus aureus and the titer was greater than 10 000 times.Conclusion: The α-HL of Staphylococcus aureus was successfully cloned and the soluble fusion protein of α-HL was successfully expressed.

Objective:To single amino acid mutation of the full human scFvs against TSLP to enhance its affinity.Methods: The specific scFvs against TSLP was screened in our previous study and here the three-dimensional structures of TSLP and anti-TSLP scFvs were simulated by Discovery Studio system,then the molecular docking was made.The amino acids of binding epitope were randomly mutated and the mutated amino acids were selected which could remarkably improve the affinity of scFvs.The primers were designed based on the sequence of mutation amino acids and the scFv sequences were mutated by the overlapping extension PCR.The DNA of mutated scFvs was ligated with the expression vector pLZ16 and transformed into E.coli DH5αF′.Then the scFvs were expressed and the scFvs with improved affinity were selected by ELISA and BIAcore.Results: The five scFvs with single amino acid mutation were screened out by DS system,which could elevate the affinity of scFvs.The mutated anti-TSLP-scFvs were amplified by PCR,which size was about 1 000 bp.The mutated scFvs with correct sequence were expressed,and the mutated scFvs with improved affinity were detected by ELISA and BIAcore.The affinity of selected mutated scFv (M4) has been about 10 times higher than the scFv nonmutation.Conclusion: The affinity of anti-TSLP-scFv has been improved successfully.

Objective To explore the factors which could lead to a preoperative diagnosis and to guide the management of cholecystoduodenal fistula (CDF).Method The experience on the diagnosis and treatment of 22 patients with CDF were retrospectively studied.These patients came from 1242 patients who received biliary tract operation in our hospital from 2001 to 2012.Results 64％ (14 of 22 patients) had 3 out of 5 of the following symptoms/signs:(1) symptoms of recurrent chills and fever,with no or only mild jaundice; (2) significant atrophy or disappearance of gallbladder on computed tomography (CT) ; (3) CT revealed complex anatomy in right upper abdomen with dilated loops of bowel; (4) ultrasound or CT revealed pneumobilia or pneumo-gallbladder; (5) barium study or duodenal endoscopy revealed obvious deformation in duodenal bulb or abnormal opening.There was no perioperative death.Morbidities included biliary fistula which presented on postoperative day 6 and day 7 in 2 patients,respectively.The daily volume of bile drainage was about 500 ml,and the biliary fistula healed after a month of conservative treatment.In addition,there were 6 patients who had infected wound,8 patients with right pleural effusion,and 8 patients with residual calculi.There was no intestinal fistula or biliary stricture.Conclusions Careful preoperative history taking and CT/uhrasound studies significantly improved the diagnostic rate of CDF.Individualized treatment reduced complications and improved clinical results.

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.

ObjectiveTo evaluate the pregnancy outcomes in female kidney transplant recipients and the long-term follow-up for the health of the offspring. MethodsClinical data from April 1978 to April 2011 of 15 female renal transplant recipients who were pregnant more than 5 months and their offspring were retrospectively analyzed. ResultsThe 15 recipients were taking CsA or Tac based immunosuppressive regimens.Twelve had successful pregnancies with stable and functioning grafts ; 1 paitent died of pulmonary infection and cardiac failure with functioning graft after the delivery of a healthy male infant; 2 experienced chronic rejection proven by biopsy at week 21 and 23 respectively,the pregnancies were therefore terminated and the grafts were lost even after rescue.All 13 newborns were smoothly delivered by cesarean section,they had an average gestational age of 35.2 ± 4.0 weeks,and a mean birth weight of 2510 ± 68 g,Apgar scale for each infant was 10,respectively.There were no birth defects,structural malformations,nor learning disabilities in 13 newborns,and their mothers all chose to bottle-feed.Thirteen children had normal intelligence,physical and mental development.Seven children experienced repeated respiratory tract infections during 0- 2 years,and 1 was diagnosed with attention deficit hyperactivity disorder.The oldest offspring is 21 years old and the youngest is 3 years old. ConclusionsFemale renal kidney recipients could achieve successful pregnancies and deliveries 3 years post transplantation with strict criteria.Their offspring were healthy during follow-up.