Objective To investigate the effects of a miRNA family member, let-7e, and a combi-nation of miR-106b and miR-20a on the expression of cytokines by THP-1 cells with Luminex xMAP technol-ogy.Methods The efficiency of transfection was evaluated by immunofluorescence assay after transfecting THP-1 cells with micrONTM mimic negative control (Cy3) for 24 h, 36 h and 48 h.The three miRNA mim-ics (let-7e, miR-106b and miR-20a) were respectively used to transfect the THP-1 cells for 24 h, 36 h and 48 h and the expression of each miRNA was analyzed by qRT-PCR analysis for screening out the optimal transfection time.The transfected THP-1 cells were stimulated with1 mg/L of LPS for 1 h.The Luminex xMAP technology was used to detect the expression of IL-8, interferon-inducible protein-10 (IP-10), mono-cyte chemotactic protein 1 (MCP-1), IL-1α, IL-6, IL-10, TNF-α, IFN-αand IFN-βin the supernatants of cell culture.A statistical analysis was performed to analyze the data obtained by using SPSS16.0 software. Results More than 90% of the transfected THP-1 cells were labeled with red fluorescence.The optimal transfection times for let-7e mimic and miR-106b/miR-20a mimics were 48 h and 24 h, respectively.Com-pared with the corresponding negative control (NC), the expression of IL-8, IP-10 and MCP-1 by THP-1 cells were enhanced after the transfection with let-7e mimic, but were inhibited after the co-transfection with miR-106b and miR-20a mimics.Conclusion The expression of IL-8, IP-10 and MCP-1 were enhanced in let-7e transfected THP-1 cells, but were inhibited in miR-106b and miR-20a co-transfected THP-1 cells.

Objective To evaluate the efficacy and safety of alemtuzumab in renal transplant recipients treated with induction therapy. Methods Eighty-nine cadaveric renal transplant recipients in our department were randomly divided into experimental group (n = 43) treated with alemtuzumab induction, 15 mg i. v. and control group (n = 46). Main immunosuppressive therapy regimen consisted of steroids, tacrolimus or cyclosporine and mycophenolate mofetil in all recipients. Post-transplant kidney function, acute rejection,infection, DGF, graft survival, lymphocyte counts were recorded within 1 year. ATP values in CD4+ T cells after transplantation was determined by using Cylex ImmuKnow assay. Results There was significant difference in the incidence of biopsy-proven acute rejection, but no significant difference was found in ImmuKnow ATP values during 6 months after transplantation and lymphocyte counts during 3 months, graft survival and the incidence of infections between the two groups. Conclusion Induction therapy with alemtuzumab appeared to be effective in the prevention of acute rejection.

Alemtuzumab (Campath) was successfully injected in 21 kidney transplant patients,7 islet transplant patients and 1 simultaneous kidney and islet transplant patient for either prevention or treatment of graft rejection.Prophylactic administration was successfully completed in all patients without discontinuation.Adverse events were not observed in 11 patients (38%),but hypertension in 18 patients (62%),shivering in 3 patients (10.3%),high fever in 3 patients (10.3%),and bronchospasm in 1 patient (3%),respectively.All complications alleviated after proper therapy.During the prophylactic administration of alemtuzumab,strict,timely and proper ward-management was needed.Care for lung,perineum,skin,diet and psychological nursing were necessary.Neither graft acute rejection nor graft chronic rejection episode occurred in all patients during 6 months to 2 years follow-up.Therefore,long term effects of Alamtuzumab and consequences of lymphocytopenia need further observation.

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 evaluate the efficacy and safety of mesenchymal stem cells(MSCs)in preventing early acute rejection after renal transplantation.Methods Eighty-eight primary cadaveric renal allograft recipients in our department were randomized into two groups treated with bone marrow MSCs (BMSCs group,n =43) or not (control group,n =45).Main immunosuppressive therapy regimen consisted of steroids,tacrolimus or cyclosporine and mycophenolate mofetil in all recipients.Estimated glomerular filtration rate (eGFR) of transplant kidney,incidence of acute reaction (AR),graft survival and incidence of adverse events were recorded within 24 months.Results In BMSCs group,the incidence of AR was 4.7 ％ and 9.3 ％ at 3rd month and 6th month respectively,significantly lower than 20.0 ％ and 26.7 ％ (P＜0.05) in the control group.The eGFR at day 7,14and 30 post-transplantation was significantly higher in the BMSCs group than in the control group (P＜0.01,P＜0.01,P＜0.05 respectively).The incidence of adverse events in the BMSCs group and the control group was 44.2 ％ (19/43) and 66.7 ％ (30/45,P ＜ 0.05) respectively and the rate of infection was 37.2 ％ (16/43) and 33.3 ％ ( 15/46,P ＞ 0.05) respectively within 24 months.Conclusion Induction therapy with autogenous BMSCs appeared to be more effective in the prevention of AR following cadaveric kidney transplantation and was associated with better clinical outcomes as far as early renal graft function without compromising patient safety.

Objective To establish a new technique of isolating pancreatic islet of langerhans and glueoeortieoid-free immunosuppressive regimen and to evaluate the clinical efficacy and safety of simultaneous adult islet-kidney transplantation in the treatment of type 1 diabetes mellitus with endstage renal failure.Methods Pancreases were stored using the"2-layer method"of the oxygenated perfluoroehemieal and UW solution.The pancreases were digested by Liberase collagenase enzyme and purified using continuous gradients of Ficoll-diatrizoic acid on a refrigerated COBE 2991 centrifuge to separate the islets.Cadaver kidney was transplanted by conventional method and cultured islets were infused by surgical approach to the liver via portal vaseulature using glucocorticoid-free immunosuppressive regimen.Clinical metabolic data such as blood glucose,dose of insulin,C-peptide,HbAlc,liver function and renal function,were determined and compared with the pre-transplant data.ResuitsIslets of langerhans were isolated successfully in 23 pancreases.The average islet yield was 300000 islet equivalents(IEQ).Islet purity and viability were 91.6%,94.6%,respectively.The stimulation index as assessing function of human islet was 3.16 and etiology results in vivo were negative.Twelve islet transplant infusions were carried out in 7 patients after kidney transplantation.Three recipients received 2 islet infusions,1 patient had 3 transplants,and 3 patients received 1 transplant only.The average islet mass for infusion was 1 1 820 IEQ/kg.The immunosuppressive regimen glucocorticoid.During 18 months to 3 yearg follow-up,4 recipients had insulin independence,the dosage of insulin decreased by 70%in 3 patients.The level of blood glucose and H bAlc,liver and renal function were normal throughout follow-up period.C-peptide of all patients was positive after islet transplantation.No adverse effects and complications related to islet infusion procedure were found.Conclusions New technique has proved tO be suitable for isolating pancreatic islet of langerhans.Simuhaneous adult islet-kidney transplantation could be used as an effective and safe way for treating type 1 diabetes mellitus with end-stage renal failure.

Primary biliary cholangitis (PBC) is a chronic cholestatic liver disease with unknown etiology, and patients with poor response to ursodeoxycholic acid and obeticholic acid may eventually progress to liver cirrhosis and even liver failure. Liver transplantation is the only effective treatment method for PBC at present. This article elaborates on liver transplantation, survival time after liver transplantation, complications, recurrence of PBC after liver transplantation, and prospects and challenges of liver transplantation in patients with PBC, so as to provide a reference for clinical outcome and treatment after liver transplantation for PBC.

Objective:To investigate the efficacy and safety of anlotinib in the treatment of advanced gastric cancer.Methods:Eighty patients with advanced gastric cancer in Longhua District Central Hospital of Shenzhen City from February 2015 to May 2016 were selected. The patients were randomly divided into anlotinib group (anlotinib 12 mg) and placebo group by random number table method. The anlotinib or placebo was given once a day for two weeks and discontinued for one week, and three weeks were a course of treatment. The relief situation, total survival time as well as adverse reactions after treatment of all patients were compared between the two groups.Results:The remission rate in the anlotinib group was higher than that in the placebo group [61.6% (37/60) vs. 5.0% (1/20)], and the difference was statistically significant ( χ2 = 19.315, P < 0.05). The overall survival time of the anlotinib group was longer than that of the placebo group [(22.8±1.0) months vs. (10.3±0.9) months], and the difference was statistically significant ( P < 0.01). The adverse reactions mainly included hypertension, diarrhea, nausea and vomiting, liver damage, etc. The adverse reactions were mild, and no drug-related deaths occurred. There was no statistically significant difference in the incidence of adverse reactions between the two groups (all P > 0.05). Conclusion:Anlotinib is effective and safe in the treatment of advanced gastric cancer.