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.
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.