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Objective To investigate the protective effect of salvianolate against bile duct injury after donation after cardiac death (DCD) liver transplantation and its clinical application prospect. Methods Sixty recipients of DCD liver transplantation were randomly divided into two groups with 30 cases in each group. Salvianolate (250mg/d) was given daily for 14 days after operation in treatment group, and the same amount of normal saline was given in control group. The therapeutic regimen of anti-rejection and anti-infection is the same between the two groups. The incidence of early graft dysfunction (EAD), and the serous levels of total bilirubin (TB), alkaline phosphatase (ALP), γ-glutamyltransferase (GGT), total bile acid (TBA) 1 month, 6 months and 12 months after liver transplantation were compared between the two groups. Also, the platelets (PLT), prothrombin time (PT), activated partial prothrombin time (APTT) and fibrinogen (FIB) were compared between the two groups at 2 weeks after operation. Results There was no significant difference in baseline parameters between the two groups (P>0.05). Compared with the control group, the incidence of EAD was decreased in treatment group, but there was no significant difference [10.0%(3/30) vs. 23.3%(7/30), P=0.166]. The serous levels of TB, ALP, GGT and TBA in treatment group were lower than those in control group 1 month, 6 months and 12 months after operation: [TB: 1 month, (28.5±17.0)μmol/L vs. (39.8±20.1)μmol/L, P=0.025; 6 months, (24.5±10.6)μmol/L vs. (33.3±16.4) μmol/L, P=0.018; 12 months, (19.8±9.5)μmol/L vs. (26.4±14.1)μmol/L, P=0.037, ALP: 1 month, (147.3±76.9)U/L vs. (187.6±70.9)U/L, P=0.039; 6 months, (163.0±61.4)U/L vs. (198.1±51.6)U/L, P=0.020; 12 months, (167.9±59.9)U/L vs. (200.2±56.2)U/L, P=0.036, GGT: 1 month, (83.9±49.5)U/L vs. (113.6±61.1)U/L, P=0.043; 6 months, (130.9±48.7)U/L vs. (169.7±77.0)U/L, P=0.023; 12 months, (154.7±45.1)U/L vs. (182.5±59.8)U/L, P=0.047, TBA: 1 month, (6.6±2.1)μmol/L vs. (8.0±2.4)μmol/L, P=0.016; 6 months, (9.5±2.2)μmol/L vs. (12.1±3.4)μmol/L, P=0.001; 12 months, (12.5±2.7)μmol/L vs. (5.6±3.8)μmol/L, P=0.001]. However, there was no significant difference in PLT, PT, APTT and FIB between two groups [PLT: (148.6±88.6)×109/L vs. (152.8±74.4)×109/L, P=0.843; PT: (12.9±1.1)s vs. (13.0±1.1)s, P=0.617; APTT: (34.6±3.7)s vs. (34.9±3.4)s, P=0.716; FIB: (3.4±0.6)g/L vs. (3.2±0.6)g/L, P=0.270, repectively]. Conclusions Salvianolate has a protective effect against bile duct injury after DCD liver transplantation, and does not increase the risk of postoperative bleeding.
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Objective To establish and improve the abdominal heterotopic heart transplantation model in mice, so as to provide references for the research on organ transplantation immunology. Methods C57BL/6 mice were selected as donors and recipients for syngeneic cardiac transplantation and recipients for allogeneic cardiac transplantation; BALB/c mice were selected as donors for allogeneic cardiac transplantation. The ascending aorta and pulmonary artery of the heart of the donors were end-to-side anastomosed with the abdominal aorta and inferior vena cava of the recipients, respectively, to establish abdominal heterotopic heart transplantation models in mice. Furthermore, the techniques of preparing recipient heart before implantation, extracting and preparing donor heart, and vascular anastomosis were improved, and the surgical results and advantages were observed and analyzed. Results Preliminary experiments were performed on 50 pairs of mice. For formal experiments, syngeneic cardiac transplantation was performed on 20 pairs and the success rate was 85.0% (17/20); allogeneic cardiac transplantation was performed on 15 pairs and the success rate was 86.7% (13/15). The total success rate of abdominal heterotopic heart transplantation was 85.7% (30/35). The average time for preparing recipient before vascular anastomosis was (11.2±2.5) min, the average time for extracting and preparing donor heart was (13.6±3.3) min, and the average time for vascular anastomosis was (21.7±3.5) min. Conclusion The improved abdominal heterotopic heart transplantation model in mouse has the advantages of easy-to-perform and high success rate, which lays a foundation for the further study of transplantation immunology.
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Objective: To observe the effect of salvianolate on the maturation and immunologic functions of mouse bone marrow-derived dendritic cells (BMDCs). Methods: Mouse BMDCs were cultured with GM-CSF and 1L-4 for 5 days to obtain immature DCs (imDCs),then the imDCs were cultured in the presence of LPS (1 μg/ml) or LPS (1 μg/ml) plus salvianolate (50, 100 and 200 μg/ml) for 48 h to obtain DCLPS , SVL , SVM and SV H. The immunophenotypes and antigen phagocytosis capabilities of imDCs, DCLPS, SVL, SVM and SVH were analyzed by flow cytometry and FITC-Dextran intake. The IL-12 p40 contents in the supernatants were determined by enzyme linked immunosorbent assay (ELISA). The stimulatory activities of imDCs, DCLPS, SVL ,SV M and SVH on allogeneic T cells in mixed lymphocyte reactions (MLR) were tested by incorporation of 3H-TdR; the contents of IL-10,IL-2 and IFN-γ in the supernatants of MLR were examined by ELISA. Results: Compared with DCLPS,SVL,SVM and SVH had lower expression of MHC-II, CD40,CD80 and CD86 molecules, secreted less IL-12 p40 in the supernatant, and exhibited stronger antigen phagocytosis capability; moreover, SVL ,SVM and SV H showed significantly weaker activity in stimulating proliferation of allogeneic T cells(P<0.05,0.01). Compared with DCLPS, SV L, SVM and SVH groups had significantly decreased IL-2 and INF-γ levels in MLR supernatants(P<0.05), but there was no significant difference in the secretion of IL-10. Conclusion: Salvianolate (50-200 μg/ml) can inhibit the maturation of mouse BMDCs in vitro, and thus has a negative effect on immunologic function of BMCDs.
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Objective: To evaluate the effect of pretreating donor liver with salvianolate on biliary injury after orthotopic liver transplantation (OLT) in rats, and to discuss the underlying mechanisms. Methods: According to the preservation solutions of the liver grafts, the recipients rats were allocated in three groups: UW solution group, salvianolate group (UW solution + 0.4 mg/ml salvianolate), and saline group (UW solution + saline). In experiment 1, the recipient rats underwent arterialized OLT with biliary extra-drainage, and the bile flow and bile salt secretion were recorded at 1, 6, and 24 h post-transplantation. In experiment 2, the recipient rats underwent arterialized OLT, and the serum alkaline phosphatase, bile salt, total bilirubin, and the incidence of biliary complications were examined at 2 weeks post- transplantation. Meanwhile, the mRNA and protein expression of bile salt export pump (BSEP) in the liver samples were also detected using real-time RT-PCR and Western blotting analysis, respectively. Results: Compared with the UW solution and saline groups, salvianolate group exhibited significantly increased bile flow and bile salt secretion at 1, 6, and 24 h post-transplantation (P<0. 05). At 2 weeks post-transplantation, salvianolate group had significantly lower serum alkaline phosphatase, bile salt, total bilirubin(P<0. 05), and incidence of biliary complications(P<0. 01), and significantly higher expression of BSEP mRNA and protein compared with the other two groups (P<0. 05). Conclusion: Salvianolate addition in UW preservation solution can ameliorate biliary injury and reduce the incidence of biliary complications after OLT in rats, which might be associated with increased expression of BSEP in liver graft.
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<p><b>OBJECTIVE</b>To investigate the effect of bilateral mandibular distraction osteogenesis in the condyles.</p><p><b>METHODS</b>16 adult hybrid dogs were randomly divided into normal control group and experiment group. Experimental dogs underwent bilateral mandibular osteodistraction at a rate of 1 min/day. 4 dogs were killed respectively in distraction period, 2 and 8 weeks after completion of 10 days distraction. The bilateral condyles specimens were harvested and examined with histological and immunohistochemical methods.</p><p><b>RESULTS</b>Compared with normal control group, various degrees of irregularities and erosion were found in fibrocartilage of condyle in experiment group, including damage in fibrous layer, hyperplasia layer and proliferative layer and osteogenic activity in cartilage layer. A significant increase of TGF-beta1 expression was also found in experiment groups. TGF-beta1 positive staining was noted in hypertrophic cell, matrix and chondroblast, osteoblast and matrix in osteogenic activity areas. These changes were the most obvious in 2 weeks after completion of distraction.</p><p><b>CONCLUSION</b>Gradual bilateral mandibular distraction at a rate of 1 mm/day brought degenerative changes of condyle, but the changes are reversible.</p>