AIM: To construct HBx eukaryotic expression vector pEYFP-C1-X and eukaryotic expression vector pGL3-P4 driven by P4 promoter of human IGF-II gene and to investigate the effect of HBx on the transcription activity of IGF-II gene P4 promoter. METHODS: HBx gene and P4 promoters were cloned into pEYFP-C1 and pGL3-basic vectors respectively by gene recombination techniques to construct recombinant plasmids pEYFP-C1-X and pGL3-P4. HepG2 cells were transfected with pEYFP-C1-X and the resistant cell clones were selected by G418. Then methylated pGL3-P4 was transiently transfected into the above cell clones, and the transcription activity of P4 promoter was determined by dual-luciferase reporter assay system. RESULTS: (1) Aim fragments HBx gene and P4 promoter that were cloned were 465 bp and 1 246 bp, respectively and the DNA sequences were accordant with GenBank data confirmed by restricted enzyme digestion and sequencing. (2) HepG2-EYFP-X cells that expressed HBx protein were obtained. (3) Luciferase activity of methylated P4 promoter in the HepG2-EYFP-X was more than that of control cell HepG2-EYFP (P<0.01). CONCLUSION: HBx may enhance the transcription activity of the P4 promoter.

Objective To compare the effects on blood lipid level by immunosuppressive drugs in renal transplantation recipients. Methods Two hundred and eighty-three renal allograft recipients with tacrolimus(FK506), cyclosporine A(CsA) and rapamycin (SRL) immunosuppressive regimen were reviewed in this study. The variation of whose total cholesterol(TC) and triglyceride(TG) concentration in serum were compared before and after three immunosuppressive regimen. Results There was no significant difference in TC and TG before and after oral FK506 for 93 patients[(4.9± 1. 1) and (1. 4±0. 8)mmol/L vs (4. 9±1.1) and (1.4±1.0)mmol/L, respectively, P>0. 05]. The concentration of TC and TG from 106 patients with CsA[(4. 8±1. 0) and (1. 6±0. 8)mmol/L vs (6. 6±1. 7) and (3. 2±1. 0)mmol/L, respectively] and 29 patients with SRL was higher than those before taking drugs, P<0. 05. The concentration was increased after 12 to 24 weeks generally. The concentration of TC and TG of CsA from FK506 to tacrolimus for 51 patients[(6. 7±1. 1) and (2. 8± 1. 0)mmol/L vs (4. 7±1. 7) and (1. 5±1. l)mmol/L, respectively] were decreased after 12 weeks (P<0. 01). Conclusions Primary factor of dyslipidemia was that CsA and SRL were used for patients post-renal transplantation, which should be regarded. The FK506-based immunosuppressive regimen should be recomended in renal transplantation patients who have a hyperlipidmia.

Objective To detect de novo development of anti-HLA antibodies after renal transplantation, and to investigate their influence on graft function. Methods 384 kidney recipients,who were negative for anti-HLA antibody before transplantation, were monitored for anti-HLA antibodies over a period of 3-96 months, and a sensitive enzyme-linked immunosorbent assay (ELISA) was used to detect anti-HLA antibodies. HLA antibody ＞10 % was defined as positive levels. Results Among 384 recipients tested, 318 recipients (82. 8 %) were negative for anti-HLA antibody after transplantation; 66 recipients (17. 2 %) developed de novo HLA antibodies, 3 recipients with HLA class Ⅰ, 61 with HLA class Ⅱ, 2 with both HLA class Ⅰ and Ⅱ. According to amino acid residue matching, 7 cases developed de novo antibodies among 92 recipients with 0 HLA-DR mismatches,compared with 59 cases among 292 recipients with 1-2 mismatches, which showed significant difference between two groups (P＜0. 01 ). 87. 4 % (278/318) recipients negative for HLA antibodies after transplantation achieved good graft function, in comparison with 65. 2 % (43/66) recipients positive for HLA antibodies (P＜0. 05). Conclusion De novo production of HLA antibodies posttransplantation may be closely associated with HLA-DR mismatch. De novo HLA antibodies posttransplantation might damage graft function and reduce graft survival rate. The detection of de novo development of anti-HLA antibodies after renal transplantation has clinical significance for assessing renal allograft function.

Objective To compare Luminex vs.ELISA methods in detecting HLA antibodies in kidney transplant recipients and their relation to acute rejection.Method Blood samples from 34 kidney transplant recipients were collected and the HLA antibodies were detected by both Luminex and ELISA methods.The sensitivity and specificity of both methods for predicting the development of acute rejection were analyzed.Results Fourteen recipients (14/34,41.17％) positive for HLA class Ⅰ antibodies were detected by using Luminex method,whereas only 1 case (1/34,2.9％) was detected with positive HLA class Ⅰ antibodies by ELISA method (P＜0.05).Similarly,13 recipients (13/34,38.24％) positive for HLA class Ⅱ antibodies were detected by using Luminex method,whereas the positive rate of HLA class Ⅱ antibodies by using ELISA method was 8.8％ (3/34,P＜0.05).The sensitivity and specificity of Luminex method for predicting the acute rejection were 80％ and 92.3％ respectively,in comparison to 30％ and 77.4％ respectively by ELISA method.Conclusion Compared to the traditional ELISA-based method,Luminex method has a better sensitivity and specificity for predicting the development of acute rejection.

Objective To analyze the clinical application of donor specific antibodies (DSAs) detected by a single antigen Luminex virtual crossmatch,and to discuss the treatment of DSA and the impact of DSA on renal function.Methods Serum from living-relative renal recipients before and after transplantation was investigated using a Luminex single antigen assay.The relation between DSA and renal acute rejection as well as renal function was analyzed.Results A total of 30 patients and 173 serum samples were tested,including 47 serum samples before transplantation,and 126 after transplantation.DSA was positive in one patient before transplantation,and 8 patients after transplantation.Three of the patients positive for DSA were treated by Bortezomib,3 by addition of MMF,2 by addition of CNI,1 by addition of Sirolimus.The MFI of DSA in one of the patients treated by Bortezomib was decreased to below 1000,while that in the other two decreased by more than 50 ％.The renal eGFR at the time with and without DSA was (1.50 ± 0.59) and (1.23 ± 0.38)ml/s respectively (P＜0.05).Conclusion Dynamic monitoring of single bead antigen antibody DSA conduces to direct the adjustment of immunosuppressant.The appearance of DSA contributes to the declination of renal function.Application of Bortezomib decreased the MFI of DSA.

Objective To study the spatial and temporal mode of infectious TB transmission in Guangxi Zhuang Autonomous Region (Guangxi).Methods Data related to infectious TB case (Include smear and/or culture positive patients) in Guangxi were collected from the National Notifiable Disease Reported System (NNDRS) from 2010 to 2015.Spatial-temporal analysis and prediction were performed by SaTScan 7.0.2,GeoDa 1.8.12,R program v 3.3.1 and SPSS 19.0 software,using the time series model,Moran' s I global and local spatial autocorrelation (Empirical Bayes adjustment).Kulldorff ' s space-time scan statistics displayed by R software was used to identify the temporal and spatial trend of TB.Results The total number of infectious TB cases,collected from NNDRS was 76 151,and showing a decreasing trend on annual incidence (value of Chi-square for Linear trend=3 464.53,P-value=0.000).The forecast value of TB cases in 2016 was 7 764 (4 971-10 557),with peak in March,analyzed through the Winters' multiplicative model.The Moran' s I global Statistics was greater than 0 (0.257-0.390).TB cluster seemed to have been existed for several years.The most significant hot spots seemed to be mainly located in the central and western parts of Guangxi,shown by local spatial autocorrelation statistics and the result from space-time scanning.Counties or districts that located in the east parts of Guangxi presented the low-low relation (significant cold spots).The situation of infectious TB seemed migratory.Conclusions Our data showed an annual decreasing trend of incidence on infectious TB with temporal concentration in spring and summer.Main clusters (hot spots) were found to be located in the central and western parts of Guangxi.Hopefully,our findings can provide clues to uncover the real mode of TB transmission at the molecular-biological level.