Risk factors for acute kidney injury after adult orthotopic liver transplantation / 中华肝胆外科杂志

Objective:To study the risk factors for acute kidney injury (AKI) after adult orthotopic liver transplantation.Methods:The clinical data of 232 recipients who underwent orthotopic liver transplantation at the First Affiliated Hospital of Zhengzhou University from January 2019 to April 2021 were retrospectively analyzed. There were 195 males and 37 females, aged (49.1±9.4) years old. The patients were divided into two groups according to whether AKI had occurred within 7 days of surgery into the AKI group ( n=112) and the non-AKI group ( n=120). Clinical data including basic information, preoperative hematological indexes, operation time and postoperative hospital stay were compared between the two groups. Factors associated with AKI after orthotopic liver transplantation were studied using univariate analysis and those factors with significant differences were included in multifactorial logistic regression analysis. Results:Among 232 patients who underwent orthotopic liver transplantation, 112 patients developed AKI after surgery, with an incidence of 48.3% (112/232). There were 64 patients with AKI stage 1 (57.1%, 64/112), 30 patients with AKI stage 2 (26.8%, 30/112), and 18 patients with AKI stage 3 (16.1%, 18/112). Logistic regression analysis showed that hypertension ( OR=5.874, 95% CI: 1.931-17.863, P=0.002) and high scores on the model for end-stage liver disease (MELD) ( OR=1.041, 95% CI: 1.010-1.074, P=0.010) were independent risk factors for AKI after orthotopic liver transplantation. Conclusion:Hypertension and MELD score were independent risk factors for postoperative AKI in orthotopic liver transplant recipients.

Strategies and outcomes of portal vein reconstruction of liver transplantation recipient with diffuse portal vein thrombosis / 中华器官移植杂志

Objective:To summarize the strategies and effects of portal vein reconstruction after liver transplantation in recipients with diffuse portal vein thrombosis(PVT).Methods:Clinical data were retrospectively reviewed for 10 PVT patients undergoing liver transplantation(LT)from January 2014 to June 2019. There were 8 males and 2 females with a age of (50.7±10.1)years. The follow-up period was (66.3±25.8)months. Diameter of portal vein anastomosis, diameter and flow velocity of portal vein and presence of ascites were evaluated by color Doppler ultrasound. And computed tomography(CT)was employed for assessing the presence of esophagogastric varices. And patency of portal vein blood flow, therapeutic outcomes of portal hypertension and survival status of recipients were evaluated.Results:Among 7 patients with diffuse PVT without enlarged collaterals, cavoportal hemitransposition( n=6) and renoportal anastomosis( n=1) were performed.Ascites subsided gradually and minimal ascites( n=4) perdisted.Variceal bleeding did not recur within 6 months.As of December 2021, portal vein blood flow remained unobstructed in 4 recipients and 3 patients died.One case of inferior vena cava thrombosis and renal injury at 3 months post operation died of multiple organ failuer at 8 months post-operation.Another patient died of recurrent hepatocarcinoma at 11 months post-operation.Another case died of stroke at 44 months post-operation.Among 3 patients with diffuse PVT complicated with enlarged collaterals, there were right gastric vein to portal vein anastomosis( n=1), gastric coronary vein to portal vein anastomosis( n=1) and pericholedochalvarix to portal anastomosis( n=1). Ascites gradually subsided within 2 months post-operation.Portal vein anastomosis thrombosis was formed 1 month after operation and racanalizated after anticogulation and thrombosis therapies in patient with gastric coronary vein to portal vein anastomosis.Upper gastroinstestinal hemorrhage occurred 36 months after operation and was relieved by endoscopic ligation and sclerotherapy.Blood flow of portal vein was unobstructed in patients with right gastric vein and pericholedochalvarix to portal vein anastomosis.During the last follow-up, velocity of portal vein surpassed 20 cm/s and liver function remained normal in 7 survivors. Conclusions:For patients with complex PVT, portal vein reconstruction may ensure sufficient portal vein blood flow of graft. After operation, portal hypertension disappears and liver function normalizes.

Correlation between the main indicators of organ donation and donor liver and the early prognosis of transplant after the death of citizens / 中华器官移植杂志

Objective To explore the correlation between main indicators of donor liver and early prognosis after liver transplantation.Methods The clinical data of 166 donors and recipients of post-mortem organ donation (DD) from June 2017 to June 2018 were retrospectively analyzed.The effects of donor age,sex,body mass index,serum sodium level,total bilirubin,prothrombin time and international standardized ratio on early allograft dysfunction (EAD) in liver transplant recipients were investigated.According to the culture results of donor liver preservation solution,the results were divided into positive group and negative group.Combined with the culture results of blood,sputum and drainage fluid after liver transplantation,the early infection rate of recipients in the two groups was observed.Results Univariate analysis showed that preoperative donor bilirubin total ＞17.1 mmol/L and donor cold ischemia time ＞8 h were risk factors for postoperative EAD in transplant recipients.Multivariate analysis showed that donor cold ischemia time ＞8 h was an independent risk factor for postoperative EAD in liver transplant recipients;the incidence of EAD in the group with cold ischemia time ＞8 h was significantly higher than that in the group with cold ischemia time ≤8 h (26.3％ vs.7.0％;P =0.003).The positive rate of postoperative sputum culture and drainage fluid culture in the donors with positive donor culture was 43.9％ and 48.8％,respectively,which was significantly higher than that in the negative group (10.7％ and 13.1％).The difference was statistically significant (P =0.000,P =0.000).The positive rate of postoperative blood culture in the positive group and the negative group was 12.2％ and 6.0％ with the difference being not statistically significant (P =0.161).Conclusion Cold ischemia time of the donor ＞8 h is an independent risk factor for EAD in recipients after liver transplantation.Shortening the cold ischemia time of donor liver can reduce the incidence of postoperative EAD in recipients.The culture results of preservation solution have a certain guiding effect on the postoperative anti-infective treatment of the recipients.

Effect and mechanism of curcumin on proliferation and apoptosis of human hepatoma HCC-LM3 cells / 国际外科学杂志

Objective To investigate the effects of curcumin on the growth and apoptosis of human hepatoma HCC-LM3 cells and explore the molecular mechanisms.Methods The human hepatoma HCC-LM3 cells were treated with different concentrations of curcumin and the cell growth inhibition rate was detected by CCK-8.The effect of curcumin on human hepatoma HCC-LM3 cells was observed.Refer to the relevant literature,the human hepatoma HCC-LM3 cells were treated with the concentration of 2.5,5.0,10.0,15.0,20.0,40.0,60.0 μmol/L of curcumin for 48 hours,taking the 0 μmol/L curcumin as control group,and the cell growth inhibition rate was detected by CCK-8.According to the results of CCK-8,selecting the concentration of 0 μmol/L as control group and the concentration of 10.0,20.0,40.0 μmol/L as experimental groups,which has significant difference on growth inhibition rates.Cell cloning assay was used to detect cell cloning ability,Flow cytometry was used to detect apoptosis,and Western blotting to detect the protein expression levels of Mcl-1,Bax,Bcl-2 and Bcl-xL.The measurement data were expressed in ((x) ± s),and the single factor analysis of variance was used for comparison between groups.Results CCK-8 assay showed that with treated by the concentration of 2.5,5.0,10.0,15.0,20.0,40.0,60.0 μmol/L,the growth inhibition rates were(6.71 ± 3.45)％,(12.33 ± 5.02)％,(20.07 ± 5.60)％,(57.80 ±7.34)％,(78.37 ±6.53)％,(91.73 ±6.14)％ and (96.18 ±3.45)％,suggesting that curcumin could inhibit the growth of human hepatoma HCC-LM3 cells in a dose-dependent manner.Cell clone formation experiment showed that curcumin could inhibit the clone of the human hepatoma HCC-LM3 cells,and the clone of the cells was inhibited significantly when the concentration of the curcumin was over 20.0μmol/L.The result of Annexin V-FITC/PI double staining analysis showed that the apoptotic rates of experimental groups and control groups were (5.20 ± 1.44) ％,(9.90 ± 3.31) ％,(55.67 ± 5.29) ％,(79.63 ±4.71)％,with all the apoptotic rates of experimental group over the control groups (P ＜0.05),suggesting curcumin could induce the apoptosis of human hepatoma HCC-LM3 cells.The Westen blotting showed that curcumin increased the expression of Bax protein while decreasing expression of Mcl-1 protein significantly in concentration-dependent manner (P ＜ 0.05),but have no effect on the expression of Bcl-2 and Bcl-xL proteins.Conclusion Curcumin could inhibit the proliferation and clone of human hepatoma HCC-LM3 cells,and induce apoptosis in a dose dependent manner.

Oridonin inhibited transplant rejection in mice / 中华器官移植杂志

Objective To investigate the role of oridonin in preventing skin graft rejection.Methods BALB/c mice were transplanted with skin grafts from C57BL/6 mice.Grafted mice were treated daily with oridonin,CsA and PBS,respectively.The survival of grafts was inspected daily and evaluated by histological analysis.On day 7 after transplantation,the percentage of CD4+ CD25+ Foxp3+ cells (Treg) in the spleen was determined by flow cytometry.The effect of oridonin on MLR and apoptosis was examined in vitro.Naive BALB/c mice were intraperitonealy injected with oridonin (15 mg/kg/day).At different time points,the number of T cells and macrophages in peripheral blood mononuclear cells (PBMCs) as well as the spleen was examined.Results The survival of skin grafts in the oridonin group (15.8 ± 1.5 days) was significantly longer than that in the control group (12.3 ± 1.2 days) and the CsA group (13.3 ± 1.1 days).Oridonin reduced inflammatory cell infiltration in grafts.The expression of Tregs was higher in the oridonin group (17.6 ± 3.6％) than in the control group (14.8 ± 2.3％).In vitro oridonin inhibited MLR and induced apoptosis in a dose-dependent manner.The number of T cells in PBMCs was rapidly decreased following oridonin treatment.With the depletion of T cells in PBMCs,high frequency of granulocytes was observed.On day 8,the number of T cells in the spleen was decreased,which was accompanied by increased phagocyte number.Conclusion Oridonin could suppress allograft rejection and prolong survival of skin grafts.The mechanism may be attributed to upregulation of Tregs and clearance of T cells.

Multivariate analysis of chondrosarcoma prognosis / 中华骨科杂志

Objective To screen possible factors affecting prognosis of chondrosarcoma.Methods A total of 37 patients with chondrosarcoma,who had undergone surgical treatment from December 2005 to March 2008 in our hospital and had complete follow-up data and definitive pathological diagnosis,were enrolled in this retrospective study.There were 16 males and 21 females,ageing from eleven to eighty-four years (average,42.8 years).The univariate analysis of survival rate was performed with Kaplan-Meier method and tested with the Log-rank test.Chi-squared test or Fisher's exact test were used to analyze numeration data,and then the significant indexes after univariate analysis were performed multivariate analysis with COX regression model to screen the independent factors affecting prognosis.On the basis of literatures,8factors including gender,age,duration of disease,tumor site,Enneking stage,surgical approach,distant metastasis and local recurrence were analyzed.Results Statistical significance was found in Enneking stage,surgical approach and distant metastasis,which indicated that they were related to survival rate 3years after surgery.However,gender,age,duration of disease,tumor site,and local recurrence had no significant correlation with prognosis.After multivariate analysis of Enneking stage,surgical approach,distant metastasis,the results showed that Enneking stage and surgical procedure were the independent prognostic factors,while distant metastasis was not an independent prognostic factor.Conclusion Enneking stage and surgical approach are the independent prognostic factors for chondrosarcoma,which can be used to evaluate prognosis of chondrosarcoma.

Construction, expression, purification and polyclonal antibody preparation of Fas extracellular region gene / 中国生化药物杂志

Purpose To construct expression vector of Fas extracellular region gene(eFas) ,to express and purify recombination protein and to prepare polyclonal antibody, which have laid a foundation of studying its function. Methods The eFas gene encoding sequence was acquired through overlapping PCR, and pET-22b ( + )/eFas expression vector was constructed. Then this vector was transformed into E. coli Rosetta-gami. Re-combinant protein was expression being induced by IPTG,and was purified using Ni-NTA matrix of affinity chromatograph. The purity of recombination protein was identified by SDS-PAGE. Hereafter, the purified eFas recombinant protein was immunized to New Zealand white rabbit in order to prepare polyclonal antibody. The titer of polyclonal antibody was determined by ELISA. Results The encoding sequence and expression vector of eFas was obtained while the interest protein was mainly expressed in the inclusion body. The eFas fusion protein's expression quantity accounts for more than 30% proportion of total E. coli protein. The eFas protein we obtained was provided with the purity of at least 95 % . Conclusion The successful constrution, expression and purification of FasL fusion protein and preparation of polyclonal antibody will provide some material for further studies of Fas.

Induction of apoptosis by Anti-DR5 mAb in synovial cells of adjuvant arthritis rats and its possible mechanisms / 中国免疫学杂志

Objective:To study the effects of Anti-DR5 mAb on inducing synovial cells of adjuvant arthritis (AA) rats and its mechanisms.Methods:AA was induced by CFA in rats.The synovial cells of rats were separated and cultured in vitro system.The apoptosis induced by anti-DR5 mAb on synovial cells was measured by MTT analysis,DNA fragmentation and flow cytometry.Caspase-3 and Bcl-2 expression in synovial cells were detected by Western blot.The involvement of the apoptotic pathway was further proved by a caspase inhibition assay.Results:MTT result showed that Anti-DR5 mAb could inhibit synovial cells growth.And flow cytometry suggested that the cell death mode was apoptosis.The protein level of caspase-3 in synovial cells treated with anti-DR5 mAb was raised,while Bcl-2 level declined.When the caspase inhibitor was added to the synovial cells treated with anti-DR5 mAb,it was showed in a dose-dependence inhibition effect,indicating that anti-DR5 mAb inducing apoptosis might be through the caspase pathway.Conclusion:Anti-DR5 mAb could induce synovial cell apoptosis through caspase pathway.And this effect may be related to the protein level of Caspase-3 and Bcl-2.

Analysis of single-chain antibody against death receptor 5 (ZF1) on murine H22 hepatocellular carcinoma cell / 中国免疫学杂志

Objective:The purpose of this study is to evaluate the effects of a single-chain antibody against death receptor 5 (ZF1) on tumor growth and survival in murine H22 hepatocellular carcinoma tumor model.Methods:Killing effect of ZF1 on H22 cells was analyzed by MTT assay in vitro. The apoptosis rate of H22 cells induced by ZF1 was detected using Flow Cytometry assay. The transplanted model of H22 tumor was developed in mice. The mice were randomly divided into four groups, PBS group, ZF1 group, EPI group and combined treatment group of ZF1/EPI. Tumor growth and body weight changes were observed. After treatment over 13 days, the tumor tissue for HE staining and TUNNEL assay was performed to detect apoptosis.Results:The results showed that ZF1 could inhibit growth of H22 cells in a dose dependent manner. The growth inhibition rate was up to 84.5%. The results showed that ZF1 alone or in combination with ZF1/EPI, the tumor growth was significantly inhibited. HE staining and TUNNEL analysis showed that ZF1 could effectively induce apoptosis of tumor cells without toxic effects, especially in ZF1/EPI combined treatment group.Conclusion:It is showed that ZF1 induces a good inhibition on the proliferation of H22 cell, especially in combined treatment group of ZF1/EPI.