Objective To explore the liver regeneration following partial liver transplantation. MethodsPartial liver transplantation in the rats were established, three experimental groups were: Ⅰ=control, partial liver resection; Ⅱ=orthotopic liver transplantation (OLT); Ⅲ=partial orthotopic liver transplantation (POLT). Liver function test, morphological investigations and liver regeneration were performed in different time after transplantation. The regenerative response of transplanted partial liver graft in rats were evaluated by Flow Cytometry and compared it to liver regeneration following resection.Results The serum concentrations of ALT, BILI increased in one week, but returned gradually to normal level within one month after transplantation. Large numbers of mononuclear cells infiltrating into the portal areas. Hepatocyte necrosis was observed on day 14 after transplantation. On day 30, the parenchyma cell showed a nearly normal appearance, bile duct proliferation was seen in portal areas. In addition, after liver resection and POLT some diploid hepatocytes were found. Dilation of the central veins, adjoining sinusoids and interlobar veins were seen in group Ⅲ. The partial liver graft is capable of regeneration similar to the situation following partial hepatectomy. The peak of liver regeneration was seen on day 1,2,4 following a hepatectomy and POLT and OLT, respectively.Conclusion The transplanted liver shows the same and/or enhanced regeneration compared to controls. There are several possible explanations for the slight delay in achieving the maximal regenerative response in rats undergoing the POLT and OLT. These may include damage that is induced by the operation itself, preservation, and reperfusion injuries. These suggest that this be caused by activation of the immune system and it might be related to the regulation of cytokines and hormone.
ObjectiveTo develop and modify a rat model of partial liver transplantation with hepatic arterialization and to investigate the postoperative histopathologic changes of the grafts. MethodsModified twocuff technique was applied to establish a rat model of partial liver transplantation, and the donor’s celiac artery was anastomosed with the recipient’s right renal artery. The biopsy of the grafts was performed on the day 1,2,4 and 7 after transplantation. Results85.4% of the recipients had survived for one week.Nuclear mitoses were seen on the postoperative day 2 by microscopy, while on the day 4 they were found occasionally. Two nucleus cells were seen on the day 4 and 7. Conclusion The stability of the model is improved by modifying the technique. Liver acupuncture biopsy is a simple, safe and feasible way to be applied in the research of postoperative problems of liver transplantation. Reconstruction of hepatic artery is possible to enhance the regeneration of liver grafts.
【Abstract】ObjectiveTo introduce a new approach for site-specific conjugation of Fab’ fragments of mouse monoclonal antibody (MoAb) to a bifunctional chelator 6-〔p-(bromoacetamido)benzyl〕-1,4,8,11 -tetraazacyclo-tetradecane-N,N′,N″,N-tetraacetic acid (BAT).MethodsFab’ fragments of B43 MoAb were obtained by a simple twostep method： 1, stable F(ab’)2 was produced by pepsin treatment of B43 MoAb. 2, Fab’ was obtained by reduction of the F(ab’)2 with cysteine. Fab’ fragments were directly conjugated with BAT. ResultsFab’ contained about 1.8 free thiol per molecule. The conjugation rate was 74% and each Fab’ fragment carried 1.28 BAT. The F(ab’)2, Fab’ and Fab’-BAT all maintained good biologic activity. ConclusionThis study may provide a simple and efficient method conjugating of MoAb Fab’ fragments to BAT.
Objective To summarize the changes of serum ceruloplasmin levels and urinary copper excretion in Wilson’s disease （WD） after living-related liver transplantation(LRLT) and orthotopic liver transplantation(OLT). Methods From September 2000 to November 2003, 140 cases liver transplantation were performed in our Liver Transplantation Center, LRLT and OLT were carried out in 26 patients with WD, three of them had fulminant hepatic failure and the others had end-stage hepatic insufficiency. All the recipients had low serum ceruloplasmin levels 〔(124.8±22.8) mg/L〕 and high urinary copper excretion 〔(1 524.8±328.6) μg/24h〕 before transplantation. The serum ceruloplasmin levels and urinary copper excretion were within normal limits in 22 donors 〔(230.4±29.6) mg/L〕 and ＜50 μg/24h〕. Results All recipients recovered satisfactorily. After operation 1, 3, 6,12 months, in OLT group, serum ceruloplasmin level and urinary copper excretion were (320.2±36.8) mg/L, (380.4±45.6) mg/L, (360.5±37.6) mg/L, (356.2±27.6) mg/L and (240.4±22.8) μg/24h, (86.5±10.6) μg/24h, (54.2±6.8) μg/24h, (46.8±3.4) μg/24h; While in LRLT group, serum ceruloplasmin levels and urinary copper excretion were (216.8±20.4) mg/L, (248.5±32.6) mg/L, (285.4±44.3) mg/L, (260.2±36.6) mg/L and (380.8±37.6) μg/24h, (150.6±24.5) μg/24h, (75.5±9.6) μg/24h, (60.3±5.8) μg/24h. Conclusion OLT and LRLT are curative procedure in WD manifested as fulminant hepatic failure and/or end-stage hepatic insufficiency. After liver transplantation, the serum ceruloplasmin level can increase to its normal range while urinary copper excretion decreases.
目的报告1例分期肝肾联合移植，并探讨分期肝肾联合移植治疗技术及其效果。方法对1例晚期乙型肝炎肝硬变伴肾功能衰竭患者实施原位肝移植术，所用免疫抑制方案为环孢霉素A与甲基强的松龙联合用药，于肝移植术后3个月行肾移植术。结果肝移植术后肝功能恢复良好，但肾功能持续恶化，经血液透析治疗无效而行肾移植术。患者肝肾联合移植术后9个月，一般情况良好，移植肝和移植肾功能均正常。结论对肝移植后各种原因导致的肾功能衰竭，当血液透析治疗无效时可再行肾移植术。同时，免疫抑制剂用量并未增加。
ObjectiveTo evaluate and summarize the perioperative management experience of living related liver transplantation (LRLT). MethodsA retrospective analysis was made in perioperative management of 13 cases undergoing LRLT (15 times operation, including 2 cases of liver retransplantation) in our department from January 2001 to December 2002. ResultsAll the operations were successful. All the 13 donors were followed up regularly, 12 donors were uneventfully recovered after operation and 1 donor suffered from bile leakage due to T tube. The survival rate of recipients, who achieved longterm survival at 2 months to 2 years, was 92.3%(12/13); the survival rate of graft was 86.7%(13/15). One adult recipient with Wilson’s disease died of serious rejection on the 72nd day postoperatively. Two cases suffered from embolism of hepatic artery, one case received reduced size cadaveric liver retransplantation, the other case received liver retransplantation from cadaver, and they both achieved longterm survival after retransplantation. The other complications included: 1 case of serious rejection, 2 cases of ARDS, 6 cases of infection of microbe, 7 cases of serious hydrothorax, 1 cases of leakage of biliary tract, and so on. ConclusionPerfect preoperative management, which composes one of the key parts of LRLT, is critically important for both donor and recipient.
【Abstract】ObjectiveTo find a stable and efficient method for ex vivo concentration of CD4+CD25+ regulatory T cells in rats. MethodsCD4+CD25+ regulatory T cells were separated from the rat splenic cells in two steps by magnetic cell sorting (MACS) system. CD4+ T cells were first negatively sorted by cocktail antibodies and antiIgG magnetic microbeads. And then CD4+CD 25+T cells were positively sorted by antiCD25 PE and antiPE magnetic microbeads. The purity and the cell survival rate of the sorted cells were measured by flow cytometry and trypan blue dyeing respectively, and the immunosuppressive action of CD4+CD25+ T cells on the proliferation of CD4+CD25- T cells was also assessed by in vitro cell proliferation assay. ResultsThe purity of negatively sorted CD4+ T cells were (83.6±2.5)% (79%~87%), and the purity of positively sorted CD4+CD25+ T cells was (90.2±1.8)% (86%~93%) with the survival rate of (92.8±3.4)% (92%~95%). These concentrated cells significantly suppressed the proliferation of CD4+CD25- T cells in mixed lymphocyte culture （CD25+/CD25- versus CD25-, P＜0.01）. ConclusionWe created a twostep procedure of magnetic cell sorting system for CD4+CD25+ regulatory T cells sorting, which insures the cells to be satisfactorily purified and well functioned.