Zhongyang Shen

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 decreased 85% in HCC, 7% in alcoholic cirrhosis and severe liver cirrhosis, along with decreases in PPARα. The 93 end-stage 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.

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), end-stage liver disease (5-fold) and HBV-related cirrhosis (3-fold). Peri-HCC also had 4-fold higher NQO1 mRNA as compared to the normal livers. GCLC mRNA levels were lower only in HCC, as compared to the normal livers and peri-HCC tissues. GCLM mRNA levels were higher in HBV-related cirrhosis and end-stage liver disease. HO-1 mRNA levels were increased in all liver tissues except for HCC. Peri-HCC had higher PRDX1 mRNA 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.

Immunogenicity of different hepatitis B virus vaccination schedules in liver transplant recipients.

To compare the immunogenicity of two modified hepatitis B virus (HBV) vaccination schedules in liver transplant recipients. Hepatitis B immunoglobulin (HBIG) in combination with nucleoside/nucleotide analogs (NUCs) is the recommended prophylaxis for preventing HBV recurrence following liver transplantation (LT). However, HBIG treatment is expensive. Active immunization with hepatitis B vaccine would be a preferable alternative prophylaxis to replace HBIG treatment. However, the overall response rate to standard vaccination (given at months 0, 1 and 6) is relatively low in immune‐compromised patients.

α-Fetoprotein is a potential survival predictor in hepatocellular carcinoma patients with hepatitis B selected for liver transplantation.

Background Risk factors can affect candidacy and prognosis following orthotopic liver transplantation (OLT) with antiviral prophylaxis for the treatment of hepatocellular carcinoma (HCC) associated with hepatitis B virus (HBV) and cirrhosis. Objective The objective of this study was to investigate the risk factors affecting OLT outcomes in patients with HCC/HBV-induced cirrhosis selected by two contemporary candidacy strategies. Patients and methods From July 2002 to December 2006, 203 patients with HCC/HBV-cirrhosis undergoing OLT with antiviral prophylaxis were evaluated retrospectively. Patients with uncomplicated HCC fulfilling Milan (conservative candidacy group) or Up-to-Seven but not Milan (inclusive candidacy group) criteria were included. Patients received postoperative immunosuppressive therapy. Tumor-free survival and overall survival (OS) were assessed. Univariate analyses between OS and clinical/demographic factors were carried out, including &agr;-fetoprotein (AFP), aspartate aminotransferase, alanine aminotransferase, tumor size, tumor nodule number, vascular invasion, lymph node metastasis, and degree of differentiation. OS was compared between the three groups on the basis of AFP level (⩽20, 20–200, and >200 ng/ml). Results Conservative candidacy group OS and tumor-free survival were better than the inclusive candidacy group. Low AST, high tumor differentiation, and low AFP were significantly associated with improved OS in the inclusive candidacy group (P<0.05). Low tumor nodule number and AFP levels were significantly associated with improved OS in the conservative candidacy group (P<0.05). AFP of more than 200 ng/ml indicated poorer outcomes in all groups. In multivariate analysis, AFP was an independent predictor of OS. Conclusion Up-to-Seven criteria may be more appropriately stratified by AFP, AST, and tumor differentiation, and AFP is a potential independent survival predictor in HBV-associated HCC patients selected for OLT.

Diabetes mellitus may affect the long-term survival of hepatitis B virus-related hepatocellular carcinoma patients after liver transplantation

AIM To determine whether diabetes mellitus (DM) affects prognosis/recurrence after liver transplantation (LT) for hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). METHODS A retrospective study was conducted between January 2000 and August 2013 on 1631 patients with HBV-related HCC who underwent LT with antiviral prophylaxis. Patient data were obtained from the China Liver Transplant Registry (https://www.cltr.org/). To compare the outcomes and tumor recurrence in the HBV-related HCC patients with or without DM, statistical analyses were conducted using χ2 tests, Mann-Whitney tests, the Kaplan-Meier method, log-rank tests and multivariate step-wise Cox regression analysis. RESULTS Univariate analysis of 1631 patients who underwent LT found overall 1-, 3- and 5-year survival rates of 79%, 73% and 71% respectively in the DM patients, and 84%, 78% and 76% in the non-DM patients respectively. Overall survival rate differences after LT between the two groups were significant (P = 0.041), but recurrence-free survival rates were not (P = 0.096). By stratified analysis, the overall survival rates in DM patients for age > 50 years (P = 0.002), the presence of vascular invasion (P = 0.096), tumors ≤ 3 cm (P = 0.047), two to three tumor nodules (P = 0.007), Child-Pugh grade B (P = 0.018), and pre-LT alanine aminotransferase levels between 40 and 80 IU/L (P = 0.017) were significantly lower than in non-DM patients. Additionally, serum α-fetoprotein level > 2000 ng/mL (P = 0.052) was associated with a significant survival difference trend between DM and non-DM patients. Multivariate analysis showed that the presence of DM (P < 0.001, HR = 1.591; 95%CI: 1.239-2.041) was an independent predictor associated with poor survival after LT. CONCLUSION HBV-related HCC patients with DM have decreased long-term overall survival and poor LT outcomes. Prevention strategies for HCC patients with DM are recommended.

Tacrolimus reversibly reduces insulin secretion, induces insulin resistance, and causes islet cell damage in rats.

OBJECTIVE To investigate the diabetogenic effects of the immunosuppressive agent tacrolimus, the reversibility of these effects upon treatment discontinuation, and the underlying mechanisms in a rat model. MATERIALS AND METHODS 60 healthy male rats were randomly divided into three groups for intragastric administration of tacrolimus either at 4 mg/kg/d or 2 mg/kg/d or an equal volume of normal saline (control). The treatment was administered for 5 months, followed by a 5-month period of no intervention. Fasting plasma glucose and insulin levels were used to calculate the homeostasis model assessment of ß-cell function (HOMA-ß) and insulin sensitivity index (ISI). RESULTS Tacrolimus treatment significantly increased blood glucose concentrations (p < 0.05) and lowered HOMA-ß and ISI (p < 0.01) in a time- and dose-dependent manner. Five months after tacrolimus treatment, significant islet cell injury was observed. However, 5 months after tacrolimus discontinuation, blood glucose concentrations significantly declined, HOMA-β and ISI levels significantly increased, and islet cell morphology noticeably improved. CONCLUSIONS In conclusion, tacrolimus treatment of healthy rats increased blood glucose concentrations in a time- and concentration-dependent manner. Development of tacrolimus-induced diabetes and reversibility after tacrolimus discontinuation may involve factors of and interactions between the insulin secretion pathway, local and/or systemic insulin resistance, and islet cell damage.

The survival benefit of liver transplantation for hepatocellular carcinoma patients with hepatitis B virus infection and cirrhosis.

Background A precise predictive survival model of liver transplantation (LT) with antiviral prophylaxis for hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC) and cirrhosis has not been established. The aim of our study was to identify predictors of outcome after LT in these patients based on tumor staging systems, antitumor therapy pre-LT, and antiviral prophylaxis in patients considered to be unfit by Milan or UCSF criteria. Methods From 2002 to 2008, 917 LTs with antiviral prophylaxis were performed on patients with HBV-cirrhosis, and 313 had concurrent HCC. Results Stratified univariate and multivariate analyses demonstrated that independent predictors for poor survival were tumor size >7.5 cm (P = 0.001), tumor number >1 (P = 0.005), vascular invasion (P = 0.001), pre-LT serum alpha-fetoprotein (AFP) level ≥1000 ng/ml (P = 0.009), and pre-LT aspartate aminotransferase (AST) level ≥120 IU/L (P = 0.044). Pre-LT therapy for HCC was an independent predictor of better survival (P = 0.028). Based on CLIP and TNM tumor staging systems, HCC patients with HBV-cirrhosis who met the following criteria: solitary tumor ≤7.5 cm, or ≤4 multifocal nodules, the largest lesion ≤5 cm and total tumor diameter ≤10 cm, or more nodules with the largest lesion ≤3 cm, and pre-LT serum AFP level <1000 µg/L and AST level <120 IU/L without vascular invasion and lymph node metastasis who were unfit for UCSF, had survival rates of 89% at 5 years. There was a 47% 5-year survival rate for patients with HCC exceeding the revised criteria. Conclusions The current criteria for LT based on tumor size, number and levels of AFP and AST may be modestly expanded while still preserving excellent survival after LT. The expanded criteria combined with antiviral prophylaxis and pre-LT adjuvant therapy for HCC may be a rational strategy to prolong survival after LT for HCC patients with HBV-associated cirrhosis.

A case of veno‑occlusive disease following liver transplantation

The present case report describes the diagnosis and treatment of a patient with veno-occlusive disease (VOD) following liver transplantation. Combining the clinical data and relevant literature, the study aimed to consider the causes of VOD following liver transplantation, and the pathogenesis, clinical diagnosis and auxiliary examination features of VOD. A 42-year-old man who had a long history of taking traditional Chinese medicine (essential components unknown) underwent an orthotropic liver transplantation on January 14, 2011, due to small venous occlusion disease of the liver. The patient was treated with tacrolimus as an antirejection therapy following the surgery, and gradually developed right upper quadrant pain and fatigue. The examination results were consistent with the diagnostic standards for VOD. Following treatment with methylprednisolone, the patient was treated with alprostadil and Danhong injections. Forty days later, the patient’s total bilirubin (TBIL) level was observed to have decreased significantly, the liver function had returned to normal and the ascites had decreased, but had not completely disappeared. The patient then underwent a transjugular intrahepatic portosystemic shunt (TIPS) procedure, following which the ascites were shown to have completely disappeared.

Anti-HBc–positive/HBsAg-negative liver donors pose a higher risk of occult HBV infection but do not cause severe histological damage in liver grafts

BACKGROUND The use of serum anti-hepatitis B core antibody (HBc)-positive/hepatitis B surface antigen (HBsAg)-negative liver donors for patients with hepatitis B virus (HBV)-related liver disease (HBRLD) is a promising means of expanding the organ donor pool and does not increase the risk of HBV recurrence. However, whether such donors will compromise the histology of the liver grafts is unclear. METHODS Among 84 patients who underwent transplantation for HBRLD and who did not have post-transplant HBV recurrence (non-detectable serum HBsAg and HBV DNA), 19 underwent liver biopsy (eight received anti-HBc-positive/HBsAg-negative liver grafts; 11 received anti-HBc-negative liver grafts) and were included in the study. Intrahepatic total HBV DNA and covalently closed circular DNA (cccDNA) were detected using real-time polymerase chain reaction; histological characteristics were evaluated with the Batts-Ludwig semi-quantitative scoring system. RESULT Of the 19 recipients, nine (47.4%) were positive for intrahepatic HBV DNA; 82.5% (7/8) received grafts from anti-HBc-positive donors and 18.2% (2/11) received grafts from anti-HBc-negative donors (P=0.003). HBV cccDNA was not detectable in the liver grafts of the 19 recipients. Ten patients had mild inflammation and minimal fibrosis in the portal area: four of the eight in the anti-HBc-positive group and six of the 11 in the anti-HBc-negative group (P>0.05). CONCLUSION Anti-HBc-positive/HBsAg-negative donors for HBRLD pose a higher risk of occult HBV infection post-liver transplant but do not cause liver damage. Thus, anti-HBc-positive grafts may be considered an effective and safe means of expanding the pool of liver donors for patients with HBRLD.

Vasculitis of anti-neutrophil cytoplasmic antibody after liver transplantation.

ObjectiveWe reported a case of vasculitis of anti-neutrophil cytoplasmic antibody after liver transplantation. A 56-year-old male patient underwent orthotopic liver transplantation of the classic style on February 25, 2010 because of alcoholic cirrhosis. MethodsPathology analysis showed it was nodular cirrhosis. The patient was given conventional treatment programs of FK (tacrolimus) + mycophenolate mofetil + hormone; hormone was suspended at the third month. He began to cough and spit, and there was blood in the sputum in September. Lung CT scanning images showed that there was a visible multiple low-density patchy shadow in both lungs; however, a number of other detecting results were negative. ResultsConsidering the patient with contact suspected mold, we improved the inspection and switched to experimental treatment (itraconazole), and the patient improved and was then discharged. Two months later, he complained about appearing hemoptysis. Final diagnostic results showed it was anti-neutrophil cytoplasmic antibody-associated vasculitis after liver transplantation. Then we stopped medication with itraconazole. Treatment with methylprednisolone was continued, and the patient gradually stopped coughing and had no expectoration and hemoptysis. ConclusionsReviewing CT respectively showed significant improvement at the 7th and 24th days of hormone therapy; thus, we confirmed it was anti-neutrophil cytoplasmic antibody-associated vasculitis after liver transplantation.