Objective To analyze the high risk factors of blood infection in Chinese citizens' organ donation,provide the basic evidence for early protection,increase the success rate of donor distribution,and expand the Chinese organ donation pool.Methods A retrospective study was performed on 70 cases of donation recruited during October 2014 to January 2016.The incidence of blood infection in these donors was analyzed.The univariate analysis and multivariate logistic regression analysis were used to find out the high risk factors influencing the donor blood infection.Finally,the donor blood infection assessment model and the receiver operating characteristic (ROC) curve were established to assess the sensitivity and specificity.Results The overall infection rate was 64.3％ (45/70).The pulmonary,blood,and urinary tract infection rate was 42.9％,31.4％ and 1.4％ respectively.The total length of hospital stay (＞10 days) (P =0.017),oxygenation index (＜ 233.5 ± 107.0) (P =0.046),aspartate aminotransferase (＞196.9 ± 329.1 U/L) (P =0.044),and valley alanine aminotransferase (＞95.0 ± 78.1 U/L) (P =0.026) were four risk factors for predicting the donor blood infection.The multivariate logistic regression analysis revealed that the total length of stay ＞10 days along with the donors' oxygenation index (＜233.5 ± 107.0) was independent risk factor for predicting the blood infection.The donor blood infection model was:0.193 + 1.753 hospital stay (＞10 days)-0.007 oxygenation index.The sensitivity and specificity were 0.682 and 0.75 (P ＜0.001) respectively.Conclusion For a long-term stay in ICU,the rate of blood infection for donors was much higher,at this time,the most effective antibiotics should be chosen.Besides,improving donor oxygenation index and liver function can reduce the incidence of infection.

Objective To explore whether polymorphisms in non-coding RNA has potential as biomarkers for predicting the risk of kidney transplantation rejection.Methods A total of 79 patients who had received kidney transplants were recruited from the First Affiliated Hospital of China Medical University and divided into the rejection group (n =26) and non-rejection group (n =53).Four polymorphisms in miRNA and 8 polymorphisms in lncRNA were detected by MALDI-TOF-MS.Results When compared with the wild genotype,the mutation genotype in miR-27a rs895819 and lnc-LRFN2-2 rs61516247 had 11.72 and 4.87 folds increased risk of kidney transplantation rejection (P =0.046,OR=1.04-131.74 and P =0.047,95％ CI =1.02-23.21,respectively).The other three polymorphisms in miRNA and 7 polymorphisms in lncRNA showed no significant associations with transplantation rejection risk (P ＞ 0.05).Conclusion The miR-27a rs895819 and lnc-LRFN2-2 rs61516247 polymorphisms were associated with the risk of kidney transplantation rejection.

Objective To explore the value of the Accordion severity grading system (ASGS) in predicting short-term outcomes after orthotopic liver transplantation for severe hepatitis by classifying post-surgery complications.Methods The clinical documents of 159 patients were retrospectively analyzed who underwent orthotopic liver transplantation for severe hepatitis between Aug.1,2004 to Sept.1,2014 at our center.Complications were categorized according to the ASGS:grade 1 (mild),grade 2 (moderate),grade 3-5 (severe),and grade 6 (death).Outcome measures included ventilator support time,the length of ICU stay,postoperative recovery time.Spearman rank correlation analysis was used to test the correlation between the different grades with these outcome measures.1-year survival trends of different grade complication groups were demonstrated by Kaplan-Meier method and compared by Log-rank test.Results In total,43 (27.0％) patients had a grade 2 complication;41 (25.8％) grade 3;31 (19.5％) grade 4;9 (5.7％) grade 5;and 35 (22.0％) grade 6.There was no grade 1 patient.There was a significant correlation between the complication grades and the ventilator support time,the length of ICU stay and postoperative inpatient time (P＜0.01).With the increase of the complication grades,the outcome measures were even worse.Severe grade complication group had a longer ventilator support time,the length of ICU stay and postoperative inpatient time than the moderate grade complication group (P＜0.01).There was a significant downward trend in 1-year survival with the increase of the complication grade (P＜0.01).Conclusion The ASGS is helpful to assess risks and predict short-term outcomes after liver transplantation for severe hepatitis.Higher Accordion grades are correlated with even worse short-term outcomes.

Objective A successful salvage treatment of acute graft-versus-host disease (GVHD) after liver transplantation(OLT) with prognosis of immune tolerance was reported and the treatment experience was summed up.Methods A 46-year-old man with hepatic carcinoma recurrence after resection underwent OLT from an ABO-identical male donor after cardiac death due to brain death.Post-transplant immunosuppression regimens consisted of induction with anti-interleukin-2 receptor monoclonal antibody (basiliximab) followed by maintenance with tacrolimus,mycophenolate mofetil and low dose of steroids.On the postoperative day (POD) 20,the patient developed skin rashes on his limbs and trunk,and skin biopsy showed histological features consistent with acute GHVD.Donor-recipient dominant HLA was matched at 6 loci,with donor CD3 + T-cell chimerism positive Results Immunosuppressants were withdrawn.Basilixirnab combined with high dose of steroids was used,and the dosage was quickly reduced.Anti-irnfection treatment was strengthened.The skin rash recovered quickly,while the hemogram was significantly decreased,which was insensitive to colony stimulating factor.The fever came back with the skin rash on the POD 46.The modified hormone regimen was used,low dose of steroids with slowly reduction,and the patient recovered with the normal hepatic function.Conclsion With the untypical clinical presentation,pathological examination,HLA-matching and chimerisms,aGVHD could be early detected and diagnosed,with a therapy of low dose of steroids with slow reduction combined with basiliximab.Recipient achieved immune tolerance,which may result from the high match of HLA and chimerisms.

Objective To observe the early effect of organ donation after pancreas-kidney transplantation.Methods Eight cases of diabetic nephropathy received combined pancreas kidney transplantation.There were 8 donors,including 6 males and 2 females,with an average age of (26 10) years old (range from 15 to 42 years).There were 4 cases of donors with China during the transition period of brain heart double death organ donation (C-Ⅲ) standard,3 cases of donors in line with the international standard of brain death organ donation (C-Ⅰ) standard,1 case of international standard of heart death organ donation (C-lⅡ M-Ⅲ) citizen donors.There were 6 men and 2 women for recipients of the same blood type.Results Eight cases were awake 4-6 h postoperation and the ventilator was removed 8-14 h after operation.The rehabilitation therapy began 2 days postoperation from surgery intensive care unit (SICU) to the common wards.Serum C-peptide and insulin levels achieved normal range in 1-2 weeks after transplant.Blood glucose returned to the normal level in 2-3 weeks,and the creatinine level decreased to the normal level in 2 weeks postoperation.Duodenal intramural hematoma occurred in one patient intraoperatively,and the pancreatic graft was removed for safe consideration.Other patients had no serious surgical complications within 2 weeks after transplantation.Conclusion For organ donation after death of pancreas kidney transplantation,early organ function recovered well.Under the strict preoperative evaluation,the young donors can be safely used in combined pancreas and kidney transplantation.

Objective To study the effect of serum uric acid (UA) levels on kidney graft function as well as long-term graft survival after renal transplantation.Methods The clinical data of 859 kidney transplant recipients from Jan.2008 to May 2014 were investigated retrospectively.The differences in clinical indexes between normal UA group and hyperuricemia group were compared based on UA levels.Cox regression model was built to analyze the effect of elevated UA on overall graft loss,death censored graft failure and death of patients,respectively.Kaplan-Meier graft survival curve was used to compare the overall graft loss,death censored graft failure and death of patients between normal UA group and hyperuricemia group.Results The average follow-up time was 38.6 ± 17.3 months for 859 kidney transplant recipients.590 (68.7％) recipients were enrolled in normal UA group and 269 (31.3％) recipients were defined as hyperuricemia patients.The average eGFR in hyperuricemia group was significantly decreased as compared with normal UA group (79.4 ± 20.93 vs.94.7 ± 20.55,P＜0.001).Cox regression model showed that if UA level increased per 10 mol/L,the risk of overall graft lost increased 1.070 times (P＜0.001) and the risk of death censored graft failure increased 1.121 times (P＜0.001) accordingly.Kaplan-Meier analysis showed the overall graft loss was dramatically decreased (P =0.009),and the death censored graft failure was significantly decreased (P＜0.0001) in hyperuricemia group as compared with that in normal UA group.The death of patients showed no significant difference between two groups (P =0.638).Conclusion Serum UA levels after kidney transplantation affect graft function as well as long-term graft survival.

Objective To observe the reception of using pig bone marrow stromal cells (BMSCs) that were transfected with human tissue factor pathway inhibitor (TFPI) to resolve the dysregulation of coagulation after liver xenotransplantation.Methods Pig BMSCs were immortalized by transfection with lentivirus containing SV40T and then transfection with human TFPI.At last the cells were tested for their ability to inhibit clotting in a model by co-incubation of human plasma,human monocytes and pig aortic endothelial cells.Results After transfection with SV40T,pig BMSCs were immortalized and similar to primary cells.The immortalized pig BMSCs showed a stable TFPI expression after transfection with human TFPI by lentivirus.Moreover,they showed the potential of regulating coagulation dysregulation in vitro.Conclusion Pig BMSCs transfected with human TFPI could solve the regulation dysregulation caused by TF activation effectively,and have the potential of resolving coagulation dysregulation in liver xenotransplantation.

Objective To study the protective effect of minocycline against ischemia-reperfusion injury after liver transplantation and its molecular mechanism after circulatory death in rats.Methods The rat donation after circulatory death (DCD) liver transplantation model was established by using magnetic-ring method.The donor and recipient were male SD rats.The rats were divided into sham operation group (SHAM group),liver transplantation group,minocycline group (MIN group),atractyloside + minocycline group (ATR + MIN group),24 rats in each group.In the MIN group,10 mg/kg minocycline was injected through the dorsum vein of the penis after reperfusion.The ATR + MIN group was injected with 2 mg/kg atractyloside.The open status of mitochondrial permeability transition pore (mPTP) was detected at 2,6,and 24 h after operation.Western blotting and immunohistochemistry were used to detect the expression of caspase3 and cyt c.Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were determined.Liver tissues were stained with hematoxylin-eosin (HE) and pathological changes were evaluated by Suzuki's standard.The survival of each group was calculated.Results As compared with liver transplantation group and ATR+ MIN group,the mPTP opening of MIN group decreased (P＜0.05),the expression of caspase3 and cyt c and the serum ALT and AST levels decreased significantly (P＜0.05),liver tissues injury was alleviated (P＜0.05),and the survival rate increased significantly after 30 days (P ＜0.05).There was no significant difference between liver transplantation group and ATR + MIN group (P ＞ 0.05).Conclusion Minocycline reduces ischemia-reperfusion injury in DCD liver transplantation in rats probably by inhibiting the mPTP opening,and preventing cyt c release and caspase3 activation to reduce hepatocyte apoptosis.This effect can be blocked by mPTP opener.

Objective To investigate the effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) on autophagy and the secretion of chemokine receptor CXCR4 induced by low-dose immunosuppressive durgs.Methods Flow cytometry was used to detect the changes of hUC-MSCs surface markers after treatment with low-dose tacrolimus and rapamycin.The effect of treatment with tacrolimus and rapamycin on proliferation of hUC-MSCs was analyzed with WST-1 assay.Regular RT-PCR was applied to analyze the mRNAs expression of ligands such as LC3B,Atg5 and Beclin1 in hUC-MSCs.Western blotting was carried out to detect the expression of LC3B,Atg5,Beclin1 and p-ULK1 in hUC-MSCs after treatment with tacrolimus and rapamycin.The secretion of chemokine receptor CXCR4 in hUC-MSCs was analyzed under the state of autophay by flow cytometry.Results Flow cytometry analysis confirmed low-dose immunosuppressive drugs tacrolimus and rapamycin did not cause changes in hUC-MSCs phenotypes significantly.Low-dose tacrolimus had no cytotoxic effect on hUC-MSCs,while,rapamycin could inhibit the proliferation of hUC-MSCs after 24 h or 48 h,with survival rate being 73.66％ and 68.81％ (P＜0.05) of controls,respectively.Moreover,both tacrolimus and rapamycin could inhibit PI3K/AKt/mTOR signaling pathway to activate hUC-MSCs autophagy,and the related proteins of LC3B,Atg5 and Beclin1 increased significantly and induced the up-regulation of CXCR4 secretion.Conclusion Our results here demonstrated that low-dose tacrolimus and rapamycin induce autophagy in hUC-MSCs and promote the secretion of CXCR4.

Objective To investigate the action mechanism of IL-35 gene transfection ameliorating cardiac allograft rejection and prolonging allograft survival.Methods pEBI3-L-p35-Fc plasmid was amplified by polymerase chain reaction.In vitro plasmid DNA pEBI3-L-p35-Fc or pSec-L-Fc was,respectively,transfected into HEK293 cells using Lipofectamine 3000.At 48 and 72 h after transfection,IL-35 concentration in culture supernatant of transfected HEK293 cells was detected by ELISA.Balb/c and C57BL/6 splenocytes treated with mitomycin (MMC) served as the stimulators,those not treated with MMC as responders,and they were subjected to one-way mixed lymphocyte culture (MLC).In the presence or absence of IL-35,the percentage of CD4+ CD25+ Tregs was detected by flow cytometry.Abdominal heterotopic heart transplantation model was established by using inbred male Balb/c mice as donors and C57BL/6 as recipients respectively.In experimental group,recipients were intravenously administrated with IL-35 plasmid (50μg) on the day 1 to day 3 post-transplantation.The control mice were treated with normal saline.The IL-35 expression in the blood,CD4+ CD25+ Tregs proportion in the blood and spleen,and the survival and the histopathologic changes of the cardiac grafts were also observed.Results In vitro the transfected HEK293 cells expressed IL-35.IL-35 enhanced the proliferation of CD4+ CD25+ Tregs of MLC in vitro.The median survival time of the cardiac grafts in experimental group (16 days) was significantly longer than in control group (7 days) (P＜0.01).As compared with control group,CD4+ CD25+ Tregs proportion was significantly increased (P＜0.01),CD8+ T cells proportion was decreased (P＜0.01) and the proliferation of lymphocytes and monocytes infiltration was inhibited in the experimental group.Conclusion IL-35 could alleviate cardiac allograft rejection and prolong cardiac allograft survival via the induction of proliferation and differentiation of CD4+ CD25+ Tregs and inhibition of proliferation of CD8 + effector T cells.