Objective To investigate the effect of regulatory dendritic cells treated by extracorporeal photochemotherapy on T cell proliferation. Methods Human peripheral blood mononuclear cells (PBMCs) were obtained and the immature dendritic cells (imDCs) were induced by recombinant human granulocyte and macrophage colony stimulating factors. SPDCs were obtained by PUVA treatment, and ECDCs were co-cultured with imDCs and PUVA-SP to obtain immunoprecipitated dendritic cells. In vitro, imDCs were co-cultured with SPDCs to obtain SPDCs; imDCs were added to 10ng/ml of LPS, and cultured for 1 day to obtain DCs. The expressions of CD11c, CD83 and CD86 on the surface of the cells were detected. The effect of imDCs on the proliferation of recipient T cells was detected by mixed lymphocyte culture method. Results The early apoptosis rate of PUVA-treated cells was 91.33%. The positive expression rates of CD83 and CD86 in ecpDCs were 22.83%±5.26% and 22.06%±4.37%, respectively, which were similar to those of imDCs (15.06%±0.59%, 15.19%±1.83% (P<0.01), but significantly lower than those in DCs (99.79%±0.36%, 99.85%±0.19%, respectively), the difference was statistically significant (P<0.01). The recipient imDC cells phagocyting the appoptotic splenic lymphocytes from the donor significantly inhibited the proliferation of recipient T cells. Conclusion Apoptosis of splenic lymphocytes induced by extracorporeal photochemotherapy can inhibit the maturation of dendritic cells and inhibit the proliferation of T lymphocytes.

Objective To evaluate the efficacy and security of splenic artery ligation for severe hypersplenism during liver transplantation.Method Thirty-two liver transplant patients with preoperative hypersplenism were selected,including 17 cases (ligation group) treated by splenic artery ligation during liver transplantation,and rest 15 patients as non-ligation group.The fluctuation of white blood cells,platelets and volume of spleen were compared between these two groups.At the same time,splenic infarction,postoperative infection,recurrent gastrointestinal bleeding,splenic artery steal syndrome and other complications were observed in these two groups.Result All recipients were followed up for over 6 months.One recipient in ligation group died of multiple organ dysfunction caused by delayed recovery of liver donor with the survival rate being 94.1％ (16/17).The survival rate in non-ligation group was 93.3 ％ (14/15) (one recipient died of respiratory failure caused by pulmonary infection).There was no statistically significant difference in survival rate between these two groups (P＞0.05).Splenic necrosis wasn't detected in the ligation group.The splenic volume in ligation group was significantly less than that in non-ligation group (P＜0.01).The products of splenic maximum length and wide diameter shrunk 33.17-± 8.26 cm2 and 22.47 ± 7.25 cm2 in ligation group and non-ligation group,respectively.The platelet counts of ligation group were significantly greater than those of non-ligation group in all the observation points within 6 postoperative months (P＜0.01).The white blood cell counts of ligation group were greater than those of non-ligation group at the first week postoperatively (P＜0.01),whereas,there was no statistically significant difference between these two groups from then on (P＞0.05).The infection incidence of ligation group was lower than that of non-ligation group within 6 postoperative months (P ＜0.05).Statistically significant differences in recurrent gastrointestinal bleeding and splenic artery steal syndrome weren't found between these two groups (P＞0.05).Conclusion Splenic artery ligation in liver transplantation is safe and effective.It can rapidly increase the counts of platelet and white blood cell in the earlier postoperative time,which is beneficial to patient's recovery.Besides,it adds no correlative complication.

Objective To analyze the correlation of tumor recurrence after liver transplantation for hepatocellular carcinoma (HCC)with the expression levels of regulatory T cell (Treg)and cytokines in peripheral blood. Methods A total of 56 patients who underwent liver transplantation in the 309th Hospital of People's Liberation Army from 2010 to 2014 were studied. According to the postoperative pathological data,all the patients were divided into the group of liver transplantation for HCC (HCC group,n=28)and group of liver transplantation for cirrhosis (liver cirrhosis group,n=28), of which the HCC group was further divided into non-recurrence group (n=8)and recurrence group (n=20)according to the situation of postoperative tumor recurrence. The expression levels of Treg and cytokines [vascular endothelial growth factor (VEGF),interleukin (IL)-2,IL-10,IL-12,transformation growth factor (TGF)-βand interferon (IFN)-γ]in peripheral blood of the patients in various groups were compared. Results Compared with the liver cirrhosis group,levels of IFN-γand IL-12 in the non-recurrence group increased significantly (both P<0.05);levels of Treg%,VEGF,IFN-γ, IL-10 and TGF-βin the recurrence group increased significantly,while levels of IL-2 and IL-12 decreased significantly (all P<0.05). Compared with the non-recurrence group,levels of Treg%,VEGF,IL-10 and TGF-βin the recurrence group increased significantly,while levels of IFN-γ,IL-2 and IL-12 decreased significantly (all P<0.05 ). Conclusions Levels of Treg and cytokines can be used to predict the tumor recurrence after liver transplantation for HCC.

Objective To explore the efficacy and safety of thymalfasin in the treatment of severe pulmonary infection after liver transplantation.Methods Twenty seven patients who developed severe lung infection after undergoing liver transplantation in Organ Transplant Institute of the 309 th Hospital of People’s Liberation Army from January 2008 to May 2014 were enrolled in this study.According to whether the application of thymalfasin,the patients were divide into thymalfasin group (n =11)and control group (n =16).In the thymalfasin group,thymalfasin was administered via subcutaneous injection at a dose of 1.6 mg once daily for consecutive two weeks.In the control group,conventional anti-infection therapy was delivered. Ventilator time,duration of fever,the length of intensive care unit (ICU)stay and mortality were statistically compared between two groups.And the incidence of acute rejection (AR)was monitored.Results Ventilator time,duration of fever,length of ICU stay of patients in the thymalfasin group were significantly shortened compared with those in the control group (all in P <0.05).There was no significant difference in the mortality between two groups.No clinical AR was observed in either group.No thymalfasin-related adverse event was found in the thymalfasin group.Conclusions Thymalfasin can improve the curative effect to anti-infection of patients with severe pulmonary infection after liver transplantation without the incidence of AR,which is efficacious and safe in the treatment of severe pulmonary infection.

Objective To discuss the relationship between Foxp3 +regulatory T cell (Treg)and tumor recurrence of patients after liver transplantation for primary hepatocellular carcinoma (HCC) over University of California,San Francisco (UCSF)criteria.Methods Clinical data of 24 patients with HCC undergoing liver transplantation in the Organ Transplantation Research Institute of the 309 th Hospital of People's Liberation Army from January 2010 to December 2013 were retrospectively studied.During the follow-up,4 patients recurred (tumor recurrence group)and other 20 patients did not recur (tumor non-recurrence group).The blood samples of healthy people was selected as control group at the same period.The levels of alpha-fetoprotein (AFP)were compared at different time points of the recurrence group and the non-recurrent group before and after transplantation.The levels of Foxp3 +Treg (Foxp3 +Treg%)were compared at different time points of the tumor recurrence group,the tumor non-recurrence group and the control group before and after transplantation.The relations between expression of Foxp3 +Treg and the levels of AFP,CD3 + and CD8 +T before and after transplantation were analyzed by correlation analysis.Results Compared with the level of Foxp3 +Treg before transplantation and the normal level,the expression of Foxp3 +Treg of patients in tumor non-recurrence group after transplantation firstly decreased,then gradually increased and finally stabilized at a low level.Compared with patients in tumor non-recurrence group,the levels of AFP and Foxp3 +Treg of patients in tumor recurrence group increased obviously and were significantly higher than the normal levels (both in P <0.01).Moreover,abnormal increase of Foxp3 +Treg at early stage was prior to AFP among the patients in tumor recurrence group.Correlation analysis indicated that the change of Foxp3 +Treg was consistent with the changes of AFP,which was positively correlated (P <0.01).But the change of Foxp3 +Treg was contrary to the change of effector T cells (CD3 +T cells and CD8 + T cells),which was negatively correlated (P <0.05-0.01).Conclusions Foxp3 +Treg is closely associated with tumor recurrence after liver transplantation for HCC.In the patients after liver transplantation for HCC over UCSF criteria,the higher Foxp3 +Treg is,the higher the recurrence risk is.Joint detection of AFP is beneficial to find tumor recurrence.

Objective To explore the influence of triple anti-tumor therapy which bases on sirolimus combined huaier granule and thymosin α-1 on T lymphocyte of rat model with liver cancer recurrence after transplantation.Methods Seventy-two Sprague-Dawley(SD)rats were randomly divided into triple therapy group,sirolimus group,huaier-granule group,thymosin α-1 group,positive-control group and blank group (n=1 2 in each group).Except the blank group,rats in all the other groups were established the simulation animal model of liver cancer recurrence after liver transplantation by chemical-induced method.After the model was established,rats in the positive control group were executed to appraise whether the model was successful.The proportion of regulatory T cells (Treg)of CD4 + T lymphocytes in peripheral blood (Treg%),the percentage of CD4 + T lymphocyte of total lymphocyte(CD4 +T%)and the percentage of CD8 + T lymphocyte of total lymphocyte (CD8 +T%),were detected by the flow cytometry respectively.The relationship between Treg% and CD4 + T %,CD8 + T %,the ratio of CD4 +/CD8 + T lymphocytes(CD4 +/CD8 +)was analyzed by the method of Spearman rank correlation.Results Pathological section of rat liver tissue suggested that the rat model was established successfully.Treg % of positive control group was higher than that of blank group,the difference had statistical significance(P <0.05).Treg% of triple therapy group was significantly lower than that of the positive control group,huaier-granule group,thymosin α-1 group,and significantly higher than the blank group (all in P <0.05 ).Compared with positive-control group,CD4 +T% and CD8 +T% of triple therapy group,sirolimus group and thymosin α-1 group were significantly higher (all in P <0.05).CD4 +T%and CD8 +T% of triple therapy group were significantly higher than those of thymosin α-1 group,sirolimus group and huaier-granule group(all in P <0.05).The relationship between Treg% and CD4 +T%,CD8 +T%, CD4 +/CD8 + in peripheral blood were negatively correlated for rats in each group.In addition,the triple anti-tumor therapy decreased the negative correlation between Treg% and CD4 +/CD8 +.Conclusions Sirolimus based triple anti-tumor therapy can decrease the peripheral blood Treg level of the liver cancer rat,increase the number of T lymphocyte and CD4 +/CD8 +,and play the role of anti tumor cell growth and proliferation.