Administration of tacrolimus in 50 liver transplantation patients

Objective: To present a good tacrolimus (FK506) administration protocol in liver transplantation. Methods: Fifty liver transplantation patients were classified into FK506 high-concentration (20 pa- tients), FK506 mid-concentration (20), CsA-FK506 (5) and FK506-CsA (5) groups. Clinical manifesta- tions, FK506 side-effects and pathological changes were observed. Results: The FK506 high-concentration group show- ed complications such as infection and liver function damage. The FK506 mid-concentration group show- ed excellent therapeutic results and some complica- tions easy to deal with. Conclusion: FK506 is a highly effective immunosup- pressive agent and the mid-concentration protocol is a better one.

Expression of P450 and nuclear receptors in normal and end-stage Chinese livers

AIM:To investigate the expression of P450 enzyme genes by using end-stage liver disease samples and trimmed normal Chinese donor livers.METHODS:The end-stage liver disease samples[n=93,including hepatocellular carcinoma(HCC),peri-HCC tissue,hepatitis B virus cirrhosis,alcoholic cirrhosis,and severe cirrhosis]and trimmed normal Chinese donor livers(n=35)from The Institute of Organ Transplantation in Beijing,China.Total RNA was extracted,purified,and subjected to real-time RT-PCR analysis.RESULTS:For cytochrome P450 enzymes 1(CYP1)family,the expression of CYP1A2 was decreased 90%in HCC,80%in alcoholic cirrhosis,and 65%in severe cirrhosis.For CYP2 family,the expression of CAR was decreased 50%in HCC,but increased 50%in peri-HCC tissues.Similar decreases(about 50%)of CYP2B6,CYP2C9,CYP2C19,CYP2D6 and CYP2E1 were observed in HCC,as compared to peri-HCC tissues and normal livers.CYP2C19 were decreased in all end-stage liver diseases and CYP2E1 also decreased in alcoholic cirrhosis and severe cirrhosis.For CYP3 family,the expression of PXR was decreased 60%in HCC,together with decreases in CYP3A4,CYP3A5,and CYP3A7.In contrast,the expression of CYP3A7 was slightly increased in HBV cirrhosis.The expression of CYP4A11 was decreased85%in HCC,7%in alcoholic cirrhosis and severe liver cirrhosis,along with decreases in PPARα.The 93 endstage livers had much higher inter-individual variations in gene expression than 35 normal livers.CONCLUSION:The expression of CYP enzyme genes and corresponding nuclear receptors was generally decreased in end-stage liver diseases,and significant differences in gene expression were evident between peri-HCC and HCC.

Activation of Aryl Hydrocarbon Receptor Prolongs Survival of Fully Mismatched Cardiac Allograft

Recent data suggest that activation of aryl hydrocarbon receptor （AhR） by its high-affinity ligand 2,3,7,8-tetrachlorodihenzo-p-dioxin （TCDD） results in expansion of regulatory T （Treg） cells and suppresses the development of autoimmune and allergic diseases in several models. Treg cells have been increasingly documented to suppress allograft rejection and even to establish stable long-term graft acceptance. However, the involvement of TCDD in the regulation of solid organ transplantation rejec- tion is largely unknown. Here, we examined whether activation of AhR with TCDD altered cardiac al- lograft rejection in an allogeneic heart transplant model. Recipient C57BL/6 （H-2b） mice were adminis- trated with a single intraperitoneal injection of TCDD, and the murine cardiac transplant models from BALB/c （H-2d） to C57BL/6 （H-2b） were built 24 h later. The complete cessation of cardiac contractility was defined as the observation endpoint. The effect of TCDD on T-cell proliferation was assessed by mixed lymphocyte reaction （MLR）. Histological and immunohistochemical analyses were performed to estimate the severity of rejection. The phenotype and cytokine profile of lymphocytes were analyzed by flow cytometry and enzyme-linked immunosorbent assay （ELISA）. Activation of AhR remarkably pro- longed the survival of cardiac allografts to more than 20 days. In vitro, TCDD ugregulated the fre- quency of CD4＋CD25＋Foxp3＋ Treg cells and suppressed the proliferation of T lymphocytes. In vivo, the prolonged survival time was associated with increased number of Treg cells in allografls and spleens Furthermore, the secretion of interferon-3, （IFN-3,） and interleukin-17 （IL-17） was reduced to less than 50% of that of the PBS treatment control group by TCDD treatment, whereas IL-10 was elevated to 10-fold of that of the PBS treatment control group. Collectively, our data indicate that activation of AhR with a single dose of TCDD significantly prolonged the survival of fully allogeneic cardiac grafts, and the mechanism underlying this effect might be involved in the induction of Treg cells.

Liver expression of Nrf2-related genes in different liver diseases

BACKGROUND： The KEAP1-Nrf2 antioxidant signaling pathway is important in protecting liver from various insults. However,little is known about the expression of Nrf2-related genes in human liver in different diseases.METHODS： This study utilized normal donor liver tissues（n=35）, samples from patients with hepatocellular carcinoma（HCC, n=24）, HBV-related cirrhosis（n=27）, alcoholic cirrhosis（n=5） and end-stage liver disease（n=13）. All of the liver tissues were from the Oriental Liver Transplant Center, Beijing,China. The expressions of Nrf2 and Nrf2-related genes, including its negative regulator Kelch-like ECH-associated protein 1（KEAP1）, its targeted gene NAD（P）H-quinone oxidoreductase 1（NQO1）, glutamate-cysteine ligase catalytic subunit（GCLC） and modified subunit（GCLM）, heme oxygenase 1（HO-1） and peroxiredoxin-1（PRDX1） were evaluated. RESULTS： The expression of Nrf2 was decreased in HCC, increased in alcoholic cirrhosis and end-stage liver disease. The expression of KEAP1 was increased in all of the liver samples.The most notable finding was the increased expression of NQO1 in HCC（18-fold）, alcoholic cirrhosis（6-fold）, endstage liver disease（5-fold） and HBV-related cirrhosis（3-fold）.Peri-HCC also had 4-fold higher NQO1 m RNA as compared to the normal livers. GCLC m RNA levels were lower only in HCC, as compared to the normal livers and peri-HCC tissues.GCLM m RNA levels were higher in HBV-related cirrhosis and end-stage liver disease. HO-1 m RNA levels were increased in all liver tissues except for HCC. Peri-HCC had higher PRDX1 m RNA levels compared with HCC and normal livers.CONCLUSION： Nrf2 and Nrf2-related genes are aberrantly expressed in the liver in different diseases and the increase of NQO1 was the most notable finding, especially in HCC.

Dysregulation of metallothionein and circadian genes in human hepatocellular carcinoma

OBJECTIVE To study the dysregulation of metallothioneins(MT)and circadian genes in achieved human hepatocellular carcinoma(HCC),Peri-HCC tissues as compared to normal human livers,which may open up a new avenue for targeting circadian clock to prevent and treatment of HCC.METHODS Total RNA was extracted,purified,and reverse transcribed.Realtime quantitative polymerase chain reaction(RT-q PCR)was performed to determine the expression of genes of metallothionein-1(MT-1),MT-2,and metal transcription factor-1(MFT-1)and circadian clock genes including core clock genes,circadian locomotor output cycles kaput(Clock)and brain and muscle Arnt-like protein 1(Bmal1),the clock feedback control genes,period(Per1,Per2)and cryptochrome(Cry1,Cry2),and clock target genes,nuclear orphan receptor factor protein(Nr1d1)and D-box-binding protein(Dbp)in human HCC,Peri-HCC and normal livers.RESULTS The expression of MT-1,MT-2,MFT-1 in human HCC was decreased,which were previously shown to have circadian rhythms.Similarly,the expression of the core clock genes(Bmal1,Clock),the clock feedback genes(Per1,Per2,Cry1and Cry2)was decreased in human HCC,as compared to Peri-HCC and normal livers.However,the expression of clock target genes(Nr1d1and Dbp)was increased in human HCC,as compared to Peri-HCC and normal livers.CONCLUSION These data clearly demonstrated the dysregulation of MTs and circadian clock genes in HCC,which could provide new insights into the relationship of circadian clock disruption and loss of MT in hepatocarcinogenesis,and could help a new strategy of targeting MT and circadian clock in the prevention and treatment of HCC.