Background Because of the lack of brain death laws in China, the proportion of cadaveric organ donation is low. Many patients with end-stage liver disease die waiting for a suitable donor. Living ...donor liver transplantation （LDLT） would reduce the current discrepancy between the number of patients on the transplant waiting list and the number of available organ donors. We describe the early experience of LDLT in the mainland of China based on data from five liver transplant centers.
Methods Between January 2001 and October 2003, 45 patients with end-stage liver disease received LDLT at five centers in China. The indication and timing, surgical techniques and complications, nonsurgical issues including rejection, infection, and advantages of LDLT in the series were reviewed. Actuarial patient and graft survival rates were calculated by using the Kaplan-Meier product-limit estimate. Statistical analysis was completed by using SPSS 10.0.
Results All LDLT recipients were cirrhotic patients, except for one man with fulminant hepatic failure. Among the 45 cases of LDLT, 35 （77.8%） were performed in one center （the First Affiliated Hospital of Nanjing Medical University）. The overall 1 and 3 year survival rate of the recipients was 93.1% and 92.0%, respectively. Of the 45 LDLT donors, there were 3 cases of biliary leakage, 2 subphrenic collections, 1 fat liquefaction around the incision and 1 biliary peritonitis after T tube removal. All donors recovered completely.
Conclusions LDLT provides an excellent approach to addressing the problem of donor shortage in China even though the operation is complicated, uncompromising and difficult with respect to the safety of the donors and receptors. Despite early technical hurdles having been overcome, perfection of technique is still necessarily. At present, LDLT is a good choice for the patients with irreversible liver disease.
Liver transplantation is indicated for Wilson's disease (WD) patients with the fulminant form and end-stage liver failure. The aim of this study was to review our experience with living-related liver ...transplantation (LRLT) for WD.
A retrospective review was made for WD undergoing LRLT at our hospital from January 2001 to February 2003.
LRLT was carried out in 15 patients with WD, one of them had fulminant hepatic failure and the others had end-stage hepatic insufficiency. The mean age of the patients was 14.5 +/- 2.5 years (range 6 to 20 years). All the recipients had low serum ceruloplasmin levels with a mean value of 126.8 +/- 34.8 mg/L before transplantation. The serum ceruloplasmin levels increased to an average of 238.6 +/- 34.4 mg/L after LRLT at the latest evaluation, between 2 and 27 months after transplantation. A marked reduction in urinary copper excretion was observed in all the recipients after transplantation. Among the eight recipients with preoperative Kayser-Fleischer (K-F) rings, this abnormality resolved completely after LRLT in five patients and partially in three. All the recipients are alive and remain well, and none has developed signs of recurrent WD after a mean follow-up period of 15.4 +/- 9.3 months (range 2-27 months) except one who died of severe rejection. The donors were 14 mothers and 1 father. The serum ceruloplasmin levels were within normal limits in all the donors (mean: 220 +/- 22.4 mg/L). The mean donor age was 35.0 +/- 4.0 years (range, 30 to 45 years). Two donors had biliary leakage and required reoperation. Grafts were harvested as follows: four right lobe grafts without hepatic middle vein and eleven left lobe grafts with hepatic middle vein. The grafts were blood group-compatible in all recipients. Two patients had hepatic artery thrombosis and underwent retransplantation.
LRLT is a curative procedure in Wilson's disease 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. Grafts chosen from heterozygote carriers do not appear to confer any risk of recurrence in recipients.
The effects of enteral nutrition containing long chain triglycerides (LCT) and medium chain triglycerides (MCT) or L‐arginine (Arg) on fat absorbability, serum free fatty acid profiles and intestinal ...morphology in rats with short‐bowel syndrome (SBS) were studied using gas chromatography. Twenty‐eight Sprague‐Dawley rats were randomly assigned to 4 groups:sham operation fed with LCT as control; 85% small bowel resection fed with LCT, MCT/LCT, and Arg/LCT, respectively. SBS rats showed a decrease of fat absorptivity. Enteral nutrition supplemented with MCT could increase fat absorptivity. L‐Arginine enhanced enteral nutrition was associated with the elevation of fat absorptivity, possibly due to its enterotrophic effect on remnant small bowel mucosa. LCT group showed a significant deficiency of total free fatty acid and the decreased essential fatty acid content, which was improved in other two SBS groups.
BACKGROUND: Glycogen synthase kinase (GSK)-3β/β-catenin signaling regulates ischemia-reperfusion (I/R)-induced apoptosis and proliferation, and inhibition of GSK-3β has beneficial effects on I/R ...injury in the heart and the central nervous system. However, the role of this signaling in hepatic I/R injury remains unclear. The present study aimed to investigate the effects and mechanism of GSK-3β/β-catenin signaling in hepatic I/R injury. METHODS: Male C57BL/6 mice (weighing 22-25 g) were pretreated with either SB216763, an inhibitor of GSK-3β, or vehicle. These mice were subjected to partial hepatic I/R. Blood was collected for test of alanine aminotransferase (ALT), and liver specimen for assays of phosphorylation at the Ser9 residue of GSK-3β, GSK-3β activity, axin 2 and the anti- apoptotic factors Bcl-2 and survivin, as well as the proliferative factors cyclin D1 and proliferating cell nuclear antigen, and apoptotic index (TUNEL). Real-time PCR, Western blotting and immunohistochemical staining were used. RESULTS: SB216763 increased phospho-GSK-3β levels and suppressed GSK-3β activity (1880±229 vs 3280±272 cpm, P<0.01). ALT peaked at 6 hours after reperfusion. Compared with control, SB216763 decreased ALT after 6 hours of reperfusion (4451±424 vs 7868±845 IU/L, P<0.01), and alleviated hepatocyte necrosis and vacuolization. GSK-3β inhibition led to the accumulation of β-catenin in the cytosol (0.40±0.05 vs 1.31±0.11, P<0.05) and nucleus (0.62±0.14 vs 1.73±0.12, P<0.05), β-catenin further upregulated the expression of axin 2. Upregulation of GSK-3β/β-catenin signaling increased Bcl-2, survivin and cyclin D1. Serological and histological analyses showed thatSB216763 alleviated hepatic I/R-induced injury by reducing apoptosis (1.4±0.2% vs 3.6±0.4%, P<0.05) and enhanced liver proliferation (56±8% vs 19±4%, P<0.05). CONCLUSION: Inhibition of GSK-3β ameliorates hepatic I/R injury through the GSK-3β/β-catenin signaling pathway.
Background Adoptive transfusion of splenocytes from long-term survivors of a tolerance model of rat orthotopic liver transplantation can induce acceptance of liver allografts in a rejection model ...preconditioned with donor γ-irradiation before liver transplantation. Recent studies suggest that the regulatory T cells (Treg cells) in splenocytes from long-term survivors play an important role in the induction of liver graft tolerance, but this observation was made from a rejection model preconditioned with donor γ-irradiation; little is known about the role of Treg cells in liver graft rejection using a naive rejection model. In this study, we examined the therapeutic potential of CD4+ CD25+ Treg cells in a naive rejection model of rat liver transplantation. Methods Freshly isolated or ex vivo alloantigen-stimulated CD4+ CD25+ Treg cells (1 × 106 cells) from naive Lewis RT1 (LEW) rats were adoptively transferred into another LEW rat on days 1 and 7 after liver transplantation from a Dark Agouti RT1a (DA) rat. Recipients were treated with or without oral tacrolimus (FK506) (0.1 mg/kg/day) from days 1 to 7 after transplantation. For ex vivo alloantigen-stimulation, CD4+ CD25+ Treg cells from LEW rats were cocultured with mitomycin C–treated DA (donor alloantigen specific) or Brown Norway (BN)(RT1n , third party) splenocytes for 72 hours. Ex vivo alloantigen-specific CD4+ CD25− T-cell proliferation responses were assessed with fresh and stimulated CD4+ CD25+ Treg cells. Results Freshly isolated, donor alloantigen-stimulated and third-party alloantigen- stimulated CD4+ CD25+ Treg cells suppressed antigen-specific CD4+ CD25− T-cell proliferation ex vivo, and adoptive transfusion of these 3 kinds of CD4+ CD25+ Treg cells prolonged survival of the liver allografts. The group transfused with the donor alloantigen-stimulated CD4+ CD25+ Treg cells had the greatest mean survival among the 3 groups (fresh Treg cells, 21 ± 2 days, n = 6; third-party alloantigen-stimulated Treg cells, 20 ± 2 days, n = 6; donor alloantigen-stimulated Treg cells, 30 ± 2 days, n = 6). When combined with short-term tacrolimus administration, adoptive transfusion of donor antigen-stimulated Treg cells induced the greatest survival time in recipients (greater than 60 days; n = 6). Conclusion Adoptive transfusion of ex vivo donor alloantigen-stimulated CD4+ CD25+ Treg cells combined with short-term tacrolimus treatment may represent a new strategy for preventing rejection after liver transplantation.
Living donor liver transplantation has been widely accepted as the treatment of choice for end-stage liver disease. Large amounts of nitric oxide generated by inducible nitric oxide synthase (iNOS) ...have been shown to play an important role in many inflammatory and immune reactions, but expression of iNOS in small-for-size liver transplantation is unknown. The aims of this study were to determine the time course of iNOS mRNA and protein as well as the redox state of liver biopsies in a rat model of small-for-size liver transplantation.
Male Sprague-Dawley rats were divided into a control group, a warm ischemia-reperfusion (IR) group, and a small-for-size liver graft group. Real-time RT-PCR and Western blotting were used to characterize the time course of the expression of iNOS mRNA and protein, respectively. Malondialdehyde (MDA) and superoxide dismutase (SOD) were used as markers to characterize the redox state of liver tissues, and the time courses of MDA and SOD levels were also measured.
The expression of iNOS mRNA and protein levels in the warm IR and small-for-size graft groups both significantly increased after reperfusion, and peaked at 3 hours. Moreover, the increase in MDA was accompanied by increased iNOS in the period of 1-24 hours after reperfusion. The MDA levels in the warm IR and small-for-size graft groups significantly increased after reperfusion, peaked at 3 hours, and decreased thereafter. The direction of change in SOD was opposite that of the change in MDA.
The expression of iNOS mRNA and protein is activated after reperfusion both in hepatic warm IR injury and small-for-size liver graft. Furthermore, the results of this study suggest that iNOS contributes to the damage in warm IR injury and small-for-size grafts via free oxygen radicals.
Methylprednisolone (MP) can affect the survival of CD4+ T lymphocytes and plays an important role in adaptive immune responses; however, its mechanism of action is not clear. Recent studies have ...shown that toll-like receptors (TLRs) on CD4+ T cells can directly modulate adaptive immune responses by affecting the survival and proliferation of activated CD4+ T cells. This study aimed to investigate the relationship between MP, TLRs and activated CD4+ T cells.
We separated and purified CD4+ T cells from mice, activated them in vitro, and co-cultured them with TLR ligands, MP or inhibitors of nuclear factor-kappa B (NF-kappaB) and activator protein 1 (AP-1). We then assessed CD4+ T cell survival and proliferation and the expression of NF-kappaB and AP-1.
Activated CD4+ T cells showed increased TLR-3 and TLR-9 mRNA expression, but polyinosinic-polycytidylic acid (poly I:C) and MP had no effect on the expression of these mRNAs. Still, poly I:C and CpG oligodeoxynucleotides (CpG DNA) increased the survival of activated CD4+ T cells, whereas MP reduced the survival of activated CD4+ T cells and could inhibit the survival effects of poly I:C and CpG DNA. The NF-kappaB essential modifier-binding domain (NBD) inhibited the survival of activated CD4+ T cells induced by poly I:C and CpG DNA, but the AP-1 inhibitor crucumin did not have the same effect. The increased expression of NF-kappaB induced by poly I:C and CpG DNA in activated CD4+ T cells could be inhibited by MP, but the same was not true for the increased expression of AP-1 induced by poly I:C and CpG DNA. Finally, the proliferation of activated CD4+ T cells was not affected by poly I:C or MP.
The survival of activated CD4+ T cells is promoted by TLR ligands, but this effect is inhibited by MP.
Portopulmonary hypertension (PPH) is clinically defined as the development of pulmonary arterial hypertension complicated by portal hypertension, with or without advanced hepatic disease. Physical ...signs may be absent in mild to moderate PPH and only appear in a hyperdynamic circulatory state. Similar signs of advanced liver disease can be observed in severe PPH, with ascites and lower extremity edema. Pulmonary hypertension is usually diagnosed after anesthetic induction during liver transplantation (LT). We present intraoperative pulmonary hypertension in a 41-year-old male patient with hepatic cirrhosis. Since this patient had no preoperation laboratory data supporting the diagnosis of pulmonary hypertension and was asymptomatic for a number of years, it was necessary to send him to the intensive care unit after operation. Further study should be focused on the diagnosis and treatment of pulmonary arterial hypertension in order to reduce its mortality.
Blockade interaction between CD28 and B7 with CTLA4Ig has been shown to induce experimental transplantation tolerance. In order to prolong the inhibitory effect of CTLA4Ig, a recombinant ...adeno-associated virus vector pSNAV expressing CTLA4Ig was constructed, and its effects on transplanted liver allografts were investigated.
The pSNAV-CTLA4Ig construct was infused into partial liver allografts of rats via the portal vein during transplantation. CTLA4Ig expression in the transplanted livers was detected with reverse transcriptase-polymerase chain reaction (RT-PCR) analysis and immunohistochemistry. Furthermore, real-time quantitative PCR was used to measure the expression of IL-2, IFN-gamma, IL-4 and IL-10 in the allografts.
The expression of CTLA4Ig in the partial allograft was detected successfully and pSNAV-CTLA4Ig improved the survival rate of rats after liver transplantation. Agarose gel analysis of RT-PCR products indicated the presence of CTLA4Ig in the pSNAV-CTLA4Ig treatment group. Cytokines expressed in allografts on day 7 after orthotopic liver transplantation showed that IL-2, IFN-gamma, IL-4 and IL-10 mRNA levels decreased in transplant recipients treated with pSNAV-CTLA4Ig compared with those treated with pSNAV-LacZ (1.62+/-0.09, 1.52+/-0.11, 1.50+/-0.07 and 1.43+/-0.07 versus 1.29+/-0.09, 1.32+/-0.07, 1.34+/-0.06 and 1.35+/-0.04, respectively).
pSNAV-CTLA4Ig effectively expressed CTLA4Ig in liver allografts. CTLA4Ig improved the pathological findings after liver transplantation. CTLA4Ig induced immune tolerance of liver transplantation, and the mechanism involved induced alteration of Th1 and Th2 cytokine transcripts. The adeno-associated virus vector encoding CTLA4Ig may be useful in the clinical study of transplantation tolerance.
To evaluate the antiangiogenic property of pigment epithelium-derived factor(PEDF) in heptocarcinoma cell lines and explore its possible application in the gene therapy of human hepatocellular ...carcinoma (HCC).
The gene encoding human PEDF was subcloned into lentiviral vector to generate the recombinant plasmid pLenti-PEDF. The plasmid pLenti-PEDF and two other packaging plasmids were cotransfected to 293T cells by calcium phosphate. Then HepG2 was infected with recombinant lentivirus and the expression efficiency of PEDF was analyzed by western blot. Proliferation and migration assay of human umbilical vein endothelial cells (HUVEC) was used to evaluate the biological activity of PEDF in vitro. Murine subcutaneous tumor model was established to investigate the therapeutic effects of Lenti-PEDF on HCC, and the expression of PEDF mRNA in tumor tissues was analyzed by RT-PCR.
Restriction enzyme digestion and DNA sequencing demonstrated that the recombinant plasmid pLenti-PEDF was constructed successfully. HepG2
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