5 cases of orthotopic live transplantation (OLTX) were performed in unresectable hepatic cellular carcinoma(3),Wilson's disease(1)and hepatitis B(1).The donor livers were harvested by the technique of rapid multiple organ harvesting,flushed and preserved with UW solution.The veno-venous bypass was introduced during the anhepatic phase in the operation.The double or triple-drug immunosuppressive regimen was used and three episodes of acute rejection were controlled in two patients.One patient died of CMV infection and another,of upper gastrointestinal hemorrhage on the 92nd and 58th postoperative day respectively.The other three are still alive for 35,150 and 210 days.

It has been shown, however, that in terms of immune function, a transitional stage of dendritic cell maturation exists between immature myeloid dendritic cells (imDCs) and mature dendritic cells (DCs), which were named semimature dendritic cells (smDCs). Therefore, the theory that DCs can be classified as imDCs and mDCs has been challenged because of finding smDCs. SmDCs and imDCs are regarded as tolerogenic dendritc cells while what concrete mechanism taken by smDCs and imDCs in transplantation immunity has yet to know. The authors analyzed the important roles and the advanced molecular mechanism of smDCs and imDCs in transplantation immunity in order to update and widen understand underlying tolerogenic dendritic cells.

BACKGROUND: During the embryonic stage, E-cadherin expression plays a critical role in the formation of hepatic tissue.OBJECTIVE: E-cadherin gene was transfected into mouse embryonic stem cells (ESCs) to observe its effects on adhesive capacity of differentiated cells.DESIGN, TIME AND SETTING: A cytological in vitro observation was performed at the Laboratory of Surgery, First Hospital Affiliated to Sun Yat-sen University between December 2007 and December 2008.MATERIALS: BALB/c mice at gestational 13 days, of clean grade, were provided by Laboratory Animal Center, Sun Yat-sen University. BALB/c mouse ESCs were preserved by professor Huang Bing from the Department of Ophthalmology, Sun Yat-sen University. CMV promoter-containing eukaryotic expression plasmid pEGFP-N1 was gifted by doctor Lu Zhi-yue from Medical College, Sun Yat-sen University.METHODS: Total RNA was extracted from BALB/c mouse fresh hepatic tissue and synthesized into cDNA by reverse transcription (RT). The synthesized cDNA was used as a template to perform a polymerase chain reaction (PCR) that amplifies a targeted fragment. Following double enzyme digestion, pEGFP-E-cadherin plasmids were reconstructed and transfected into mouse ESCs. In vitro differentiation of transfected mouse ESCs was performed.MAIN OUTCOME MEASURES: Detection of E-cadherin expression in the differentiation system using RT-PCR and immunocytochemistry and observation of adhesive capacity of differentiated cells.RESULTS: E-cadherin gene-transfected ESCs could stably express E-cadherin during differentiational 1-17 days, while non-transfected ESCs expressed a decreasing amount of E-cadherin. The adhesive capacity of differentiated cells that stably expressed E-cadherin was markedly enhanced. Compact cell connection and multi-layer growth state remained at 19 days. While non-transfected ESCs gradually changed from embryoid bodies into noncohesive cell populations.CONCLUSION: Differentiating E-cadherin ESCs exhibit markedly enhanced adhesive capacity and maintain multi-layer growth state.

BACKGROUND:E-cadhedn plays an important role in development of liver tissue in the embryonic stage.Therefore,it is importance for investigating the feasibility of dynamic expression of E-cadherin in embryonic stern cell differentiation to in vitro development of liver tissue.OBJECTIVE:To observe the dynamic expression of E-cadherin in embryonic stem cell differentiation and the effect on cell adhesion.DESIGN,TIME AND SETTING:An in vitro cytological observation was performed at Surgical Laboratory of the First Affiliated Hospital of Sun Yat-sen University from December 2007 to December 2008.MATERIALS:Embryonic stem cells of BALB/c mice were obtained from Professor Huang (Department of Ophthalmology,Sun Yat-sen University).Twenty 13-day-old pregnant BALB/c mice of clean grade were provided by the Experimental Animal Centar of Sun Yat-sen University.METHODS:Following trypsinization,embryonic stem cells were suspend-incubated in DMEM culture medium containing fetal bovine serum,2-mercaptoethanol,HEPES,penicillin,and streptomycin.Embryoid body was formed 5 days after normal development and incubated in the culture plate at day 6.Liver tissue which was obtained from 13-day-old pregnant BALB/c mice was prepared for fetal liver cells which were frozen-sectioned as the controls.MAIN OUTCOME MEASURES:E-cadherin expression was detected using reverse transcriptase-polymerase chain reaction (RT-PCR) and immunocytochemistry st varying time points of 1,5,9,13,and 17 days in stages of initial differentiation of embryonic stem cells,formation of embryoid body,and formation of differentiated clusters.Additionally,the effect of E-cadherin expression on cell adhesion was also detected.RESULTS:RT-PCR showed that E-cadherin mRNA expression was not observed at day 1 but peaked at day 5;gradually,the expression was decreased until the expression was stopped at day 17.E-cadherin mRNA expression was strong in fetal liver cells in the control group.Immunocytochemistry showed a similar outcome.Morphologically,embryonic stem cells developed from unicells into compact three-embryonic layer embryoid body and into incompact cell population.CONCLUSION:E-cadherin expression correlates with differentiated cell adhesion;additionally,the lost expression in an in vitro environment may be an important cause for unable regularization of differentiated cells.

Objective To evaluate the effect of single injection of low dose zoledronic acid on bone resorption.Methods332 menopausal patients with bone deficiency treated in our hospital were selected.The patients were treated with zoledronic acid 1mg (1mg group), 2.5mg treatment group (2.5mg group), 5mg treatment group (5 mg group) and placebo treatment group (control group), each group of 83 patients.The patients of 1mg group, 2.5mg group and 5 mg group were treated with 1mg, 2.5mg and 5mg zoledronic acid alone.The patients in the control group were given intravenous infusion of placebo.Evaluated the lumbar spine (L1-L4) and total hip bone mineral density (BMD) and bone metabolic markers at baseline, 6, 12, 18, and 24 months in the four groups.The bone metabolic criteria included t β-Cterminal-telopeptide of type I collagen (β-CTX) and procollagen type-I N-terminal propeptide (P1NP).ResultsThe Lumbar spine BMD and the Total hip BMD were significantly higher in 1mg group than baseline value and Simultaneous valueand in the control group (P<0.05), The difference were statistically significant (all P<0.05).The values at 8 and 24 months decreased gradually.The value was significantly lower (P<0.05) compared with the control group, There were no statistically significant difference compared with the simultaneous value in control group.The lumbar BMD and the total hip BMD in 2.5mg and 5mg groups were significantly lower than the baseline values during the whole trial period (all P<0.05).The trend of β-CTX and P1NP was similar to that of BMD in each group.ConclusionIntravenous injection of 1 mg and 2.5 mg of zoledronic acid produces anti-bone resorption that can last for at least 1 year.After one year of treatment, The effect of single injection of 2.5 mg of zoledronic acid on bone is similar to that of single injection of 5 mg zoledronic acid.1 mg zoledronic acid produced by anti-bone resorption can last for 12 months, and then slowly disappear.

Objective To investigate the relationship between the intercellular adhesion molecule-1(ICAM-1) and lymphocyte function associated with antigen-1( LFA-1) and acute rejection of liver transplantation in rats. Methods SD rats and Wistar rats were randomly divided into normal control group, isograft group, and allograft group. Liver biopsies from the recipients were obtained on the day 2,4, and 7 postoperatively to examine the expression level of ICAM-1 and LFA-1 in liver grafts. Samples were firstly stained with specific monoclonal antibodies using immunohistochemistry stain and then applied to image semiquantitive analysis. Results The expression levels of ICAM-1 and LFA-1 in allograft group were obviously higher than those in both isograft group and normal group. Moreover, the expression levels of ICAM-1 and LFA-1 were correlated positively with the degree of acute liver rejection. Conclusions Detecting the expressions of ICAM-1 and LFA-1 in liver graft tissue may be helpful to the diagnosis of acute rejection after liver transplantation.

【Objective】To investigate the relationship between the expression of lymphocyte function-associated antigen-1 (LFA-1) on peripheral lymphocytes and acute rejection after orthotopic liver transplantation (OLTx) in rat.【Methods】Adult male rats were divided into 2 groups.In the non-rejection group,40 SD rats were used as both donors and recipients.In the rejection group,20 Wistar rats were used as donors and 20 SD rats as recipients.Blood samples were collected through the tail vein 1 day before transplantation and on days 1、3、5、7 after OLTx.The expression of LFA-1 (CD11a) on peripheral lymphocytes was analyzed by using indirect immunofluorescent marker-flow cytometry.【Results】The expression level of LFA-1 on peripheral lymphocytes in recipient rats after orthotopic liver transplantation was markedly lower than that before operation (P<0.01);The expression level of LFA-1 on peripheral lymphocytes in rats with acute liver rejection was significantly higher than that in the non-rejection group (P<0.01).【Conclusion】Monitoring the expression level of LFA-1 on peripheral lymphocytes may be helpful to the diagnosis of acute graft rejection.

Objective To investigate the quality of life and its influencing factors in hver transplantation recipients in order to supply scientific reference for improving their life quality.Methods 33 patients with terminal liver diseases waiting for liver transplantation(the pre-operation group)and 102 patients 2 weeks after liver transplantation and outpatients for follow-up visits(the post-operation group)were conducted with SF-36,self-rating anxiety scale,self-rating depression scale,and social support questionnaire.Results Except for the scores of physiological function,emotional role,social function in SF-36.differences of scores of other factors were evident between the preand post-operation groups.Life quality of patients over 1 year after operation was better than those within 1 year. The influencing factors of QOL were subjective support and anxiety.Conclusions After liver transplantation,the patients' QOL evidently improves.Life quality of patients over 1 year after operation was better than those within 1 year.Patients' QOL is positively related to subjective support,negatively related to anxiety.

OBJECTIVETo explore the diagnosis, treatment and long-term outcome of late acute rejection (LAR) following adult orthotropic liver transplantation (OLT).

METHODSA total of 398 consecutive adult patients who underwent OLT in Organ Transplant Center, the First Affiliated Hospital, Sun Yat-sen University between January 2007 and December 2012 were reviewed retrospectively. There were 48 patients (12. 1%) developed to LAR, including 43 male patients and 5 female patients, with an average age of (52 ± 13) years(18 - 70 years). The mean body mass index was (22.1 ± 4. 5) kg/m2 (15. 4 - 30. 4 kg/m2). The indications of the liver transplantation recipients included 16 cases of end-staged liver cirrhosis after hepatitis B or C(33. 3%), 14 cases with severe hepatitis (29. 2%), 9 cases of primary liver cancer(18. 5%), 5 cases of alcoholic liver cirrhosis (10. 4%), 1 case with autoimmune liver disease (2. 1%) , the other 3 cases (6. 3%). They were followed up by outpatient service, telephone and other means. Survival curves were generated with the Kaplan-Meier method and Cox proportional hazards modeling was used for predictors of mortality. Statistically significant variables found by single factor regression analysis were put into the Cox proportional hazards regression model of multivariate analysis.

RESULTSThe time-to-event was 23. 6 months after OLT which were more common in the first year to the third year post-transplant (26/48,52. 4%). Thirty-five cases were assessed as mild, 11 cases were assessed as moderate, and 2 cases were assessed as severe ,based on the Banff schema. After adjustment to the immunosuppressive regimen, the overall recovery rate reached to 81. 3%. The rate of steroid-resistant acute rejection was 11. 8% (4/34). Inadequate immunosuppression and steroid pulsation were two independent risk factors affecting the prognosis of LAR (P = 0. 008, P = 0. 003, respectively).

CONCLUSIONSLAR is an uncommon complication after OLT. Inadequate immunosuppression and steroid pulsation are the major risk factors for prognosis of LAR. Improving patient compliance and strengthening blood concentration surveillance can increase the patient survival.

Acute Disease ; Adolescent ; Adult ; Aged ; Female ; Graft Rejection ; diagnosis ; mortality ; therapy ; Hepatitis B ; Humans ; Immunosuppression ; Liver Cirrhosis ; Liver Neoplasms ; Liver Transplantation ; mortality ; Male ; Middle Aged ; Multivariate Analysis ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Risk Factors ; Steroids ; Young Adult

BACKGROUND:Nowadays,liver grafts for transplantation are clinically sourced from non-heart-beating donors.Moreover,there is still no uniform determination of safe time limit points for non-heart-beating donor liver in warm ischemia and cold preservation.OBJECTIVE:To evaluate the application safety and curative effects of warm ischemia liver graft affected by varied cold ischemia time (CIT) in liver transplantation.DESIGN,TIME AND SETTING:A randomized controlled observation was performed in the Organ Transplantation Center,First Affiliated Hospital of Sun Yat-sen University between January 2006 and December 2007.PARTICIPANTS:154 cases who underwent non-heart-beating liver transplantation were included in this study.All liver graftsinvolved had a warm ischemia time(WIT) less than 10 minutes.METHODS:All cases were assigned into 3 groups according to CIT:group Ⅰ (CIT＜8 hours,n=58),group Ⅱ (CIT 8-12hours,n=62),and group Ⅲ (CIT 13-16 hours,n=34).The liver grafts were randomly allocated for the patients.Following liver transplantation,the same immunosuppression protocol was employed for each group.MAIN OUTCOME MEASURES:Following surgery,peak level of alanine aminotransferase (ALT),primary graft dysfunction (PGD) after liver transplantation,acute rejection response,biliary complications,vessel complications,perioperative infections,graft and recipient survival rate were compared among 3 groups.RESULTS:Follow-up time was 8-32 months.No PGD was detected in all 154 cases.Group Ⅱ showed postoperative ALT peak levels significantly higher than group Ⅰ (P＜ 0.05).There was no significant difference between groups Ⅰ and Ⅱin terms of acute cellular rejection,perioperative infection,biliary complication,vessel complication,graft survival rate,and recipient survival rate (P＞0.05).Compared with group Ⅰ,the group Ⅲ exhibited significantly increased postoperative ALT peak level,biliary complications,and perioperative infections,and significantly decreased graft and recipient survival rate (P＜0.05).CONCLUSION:Non-heat-beating-liver grafts with less than 10 minutes of warm ischemia can tolerance 12 hours of cold ischemia.More than 12 hours,postoperative liver transplantation complications ascend and,contradictorily,graft and recipient survival rate descend.