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BACKGROUND:Hepatocyte transplantation has attracted more and more attention as a therapeutic measure for liver failure and genetic metabolic liver diseases.OBJECTIVE:TO evaluate the efficacy and safety of human hepatocyte transplantation in treating hepatitis B related liver failure in one case by a 2-year follow-up.DESIGN:A case-report of 2-year follow-up.SETTING:No.9 Department of Infectious Diseases,Bioengineering Research Room,the 302 Hospital of Chinese PLA.PARTICI PANT:One inpatient with hepatitis B related liver failure was selected from the 302 Hospital of Chinese PLA.and she was diagnosed according the laboratory tests.The transplanted hepatocytes were originated frOm the healthy liver of a 24-year-old man,who had signed the protocol for liver donation before death.METHODS:The hepatocyte transplantation was completed in the Department of Radiology,the 302 Hospital of Chinese PLA in December 2004.Liver was isolated to obtain human primary hepatocytes, and then cryopreserved.The hepatocytes were transplanted into recipient spleen via femoral vein after resuscitation.The clinical symptoms,changes of blood biochemical indexes,and changes of spleen MRI signals were observed before and after operation.The patient was reexamined every half a year after operation, including liver function, blood coagulation function,B-mode ultrasonography,gastroscopy and MRI,and she was followed up for 2 years. MAIN OUTCOME MEASURES:Liver function,blood coagulation function, imaging indexes, immunological indexes,complication and rejection.RESULTS:①Totally(1-2)×1010 hepatocytes were harvested,and the viability of rewarmed hepatocytes was 60%,and finally 2×109 hepatocytes were transplanted.②Two months later,the clinical symptoms of the recipient were obviously ameliorated,and serum bilirubin and aspartate aminotransferase(AST)were obviously decreased,while prothrombin activity was markedly increased.20 months later,the MRI results showed that there was hepatocyte image in spleen.Two years after operation.the total bilirubin level was 20 μmol/L,direct bilirubin level was 7 μmol/L, alanine aminotransferase was 416.75 nkat/L,AST was 533.44 nkat/L,albumin was 37 g/L,prothrombin activity was 90%,which were all obviously ameliorated as compared with those before operation(474.5 μmol/L,340.3 μmol/L,400.08 nkat/L,1 200.24 nkat/L,38 g/L,25%).The patient left the hospital 2 months later and could do light-burdened job.No complications of hydroperitonia and liver function failure, etc.were observed,and no rejection occurred.Several reexaminations by B-mode ultrasonography all indicated the further aggravations of liver cirrhosis and esophageal varices.She was admitted to hospital for twice because of esophageal varices bleeding,and cured by endoscopic variceal sclerosis therapy.CONCLUSION:Hepatocyte transplantation can ameliorate liver function without rejection,but it cannot relieve portal hypertension.
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Objective To study the changes in PAF and its receptor in Kupffer cells in experimental cirrhosis and to evaluate the role of activated Kupffer cells in portal hypertension. Methods Kupffer cells, isolated from the livers of control and CCl_4-induced cirrhotic rats, were cultured in serum-free medium overnight. PAF synthesis and release by Kupffer cells were determined 24 h later by rapid ~3H-PAF scintillation proximity assay, and the expression of PAF receptor in Kupffer cells by saturation binding technique and semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). By immunohistochemisty, the distribution of PAF in Kupffer cells was surveyed. Results Cell-associated PAF synthesis and release was increased about 1.48 fold and 2 fold, respectively, by Kupffer cells in cirrhotic liver as compared with the control (P0.05). Consistent with the receptor binding capacity, the mRNA expression of PAF receptor increased significantly in the Kupffer cells of cirrhotic liver (P
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Objective To investigate the correlation between Kupffer cells and the synthesis of platelet activating factor(PAF)in experimental hepatic cirrhosis,and to elucidate the effects of endothelin(ET)on portal hypertension.Methods Thirty SD rats were randomly assigned to two groups:control group and CCl4-induced hepatic cirrhotic group.Kupffer cells,isolated from the livers of animals in both groups,were cultured for 24h.ET-1-induced PAF synthesis,and mRNA expression of PAF,ET-1 receptor and preproendothelin-1 in Kupffer cells were determined by rapid 3H-PAF scintillation proximity assay,saturation binding technique and semi-quantitative reverse transcriptase polymerase chain reaction(RT-PCR),respectively.Results Cell-associated PAF synthesis and release increased about 1.48 folds and two-folds,respectively,by cirrhotic Kupffer cells as compared to the control(1.02 ? 0.06 vs 0.69 ? 0.07 pg /mg DNA in Kupffer cells and 1.42? 0.14 vs 0.66 ? 0.04 pg/mg DNA in medium).Endothelin-1 enhanced Kupffer cells to stimulate PAF synthesis in a concentration-dependent manner,and for cirrhotic Kupffer cells,the effect was more significant than control.Cirrhotic Kupffer cells also had increased densities of functional receptors for both PAF and ET-1(exclusively ETB),but did not change the affinity of these receptors.No mRNA transcripts for the ETA receptor or preproET-1 were detected.Conclusion Kupffer cell is the main source of PAF in the cirrhotic rats.ET-1 stimulates PAF synthesis in activated Kupffer cells via ETB receptor.Since both ET-1 and PAF individually cause portal hypertension,Kupffer cells may play a role in portal hypertension associated with liver cirrhosis.