BACKGROUND: It has been suggested that obesity and fatty liver may be associated with the morbidity and mortality of liver cancer, and the early diagnosis and effective treatment of fatty liver coupled with liver cancer are supposed to improve the prognosis of obese patients. This review was attempted to understand the relationship between obesity, fatty liver and liver cancer.
DATA RESOURCES: An English-language literature search using PUBMED (1990-2004) on obesity, fatty liver and liver cancer and other related articles in Chinese.
RESULTS: Obesity is associated with the risk of death from all cancers and from cancers at individual sites including liver cancer, and it is an independent risk factor for hepatocellular carcinoma (HCC) in patients with alcoholic cirrhosis and cryptogenic cirrhosis. Because nonalcoholic steatohepatitis has been implicated as a major cause of cryptogenic cirrhosis, the development of HCC may be part of progressive nature of this condition.
CONCLUSIONS: Obesity is associated with the incidence and mortality of HCC. More frequent surveillance for HCC may be warranted in obese patients with fatty liver and attempts should be made to interrupt the progression from simple hepatic steatosis to steatohepatitis, cirrhosis and ultimately HCC.

BACKGROUND: Paraoxonase 1(PON1) is an ester hydrolase in serum and in the liver. Studies have suggested that PON1 measurement to the current battery of tests may improve the evaluation of chronic liver diseases. The aim of this study was to investigate the clinical significance of monitoring the level of serum PON1 activity in liver transplantation patients.
METHODS: A series of biochemical indexes were monitored in preoperative, operative and postoperative serum samples of 17 liver-transplanted patients. The change of serum PON1 level and its relations with other biochemical indexes were analyzed.
RESULTS: PON1 was distributed normally in the healthy population and its reference value ranged from 45.5 to 265.8 U/mL. The PON1 level of all patients was lower than that of control group significantly (P<0.001); the level began to elevate continuously 5 minutes after opening of the portal vein and was higher than that 90 minutes after opening of the portal vein (P<0.05). Two days after operation it was still higher than the normal. The levels of serum ALT and AST elevated more significantly after opening of the portal vein than before operation and they were higher than the normal values till 2 days after the operation.
CONCLUSIONS: The level of PON1 in serum may be taken as one of the effective indexes to assess whether the implant is alive and to monitor liver function of the patient together with other tests.

BACKGROUND: The most common complications after liver transplantation are acute and chronic rejection. Although special changes can be found clinically and pathologically in both of them, sometimes other complications such as preservation injury, recurrent disease (HBV) and hepatic artery thrombosis should be differentiated from them.
METHODS: Five patients with various complications after liver transplantation were synthetically analyzed according to their clinical information, laboratory tests and liver biopsy findings so as to find out the clinical rules and the histological features.
RESULTS: Special changes and diagnostic standards were found clinically and pathologically in both acute and chronic rejection. Other complications such as fibrosing cholestatic hepatitis and hepatic artery thrombosis also had their own pathological features. Evaluation of donor liver should be done before liver transplantation.
CONCLUSION: It is very important to understand and master the diagnostic standards of rejection and the major points for its differentiation.

BACKGROUND: Over 355 patients have received orthotopic liver transplantation (OLT) at this hospital since 1993. Preoperative imaging studies of both hepatic vessels and parenchyma in these recipients bettered surgical planning or even precluded the necessity of surgery. Here we report our preliminary results of modified magnetic resonance angiography (MRA) using sensitivity encoding (SENSE) through comparative study with conventional digital subtraction angiography (DSA) and CT arterial portography (CTAP). 
METHODS: Sixteen patients with suspected liver diseases were included in the study. All of them received both dynamic MRI of the liver using SENSE and digital DSA with CTAP within a two-week interval. The four-phase MRA was reconstructed from source images of the coronal dynamic study. The arterial phase of the modified MRA was compared with DSA in the evaluation of hepatic arteries and the portal phase compared with CT portography reconstructed from source images of CTAP. In dynamic study of the liver, a fixed dose (20 ml) of contrast medium and scan timing were used.
RESULTS: The main branches and variations of the hepatic arterial system were well shown on the modified MRA, although the marginal branches of hepatic arteries were of poor quality. The figures of portal veins on MRA were as clear as or superior to those of CTAP. In addition, the suprarenal inferior vena cava (IVC) was well demonstrated on MRA and/or non contrast-enhanced coronal balanced fast-field echo (b-FFE) scan sequence in most cases. MRI detected most parenchymal lesions of the liver and hemodynamics of these lesions could be evaluated on source images of the modified MRA. MRI/MRA also serendipitously revealed several extrahepatic disease entities or variations that were not found on DSA/CTAP.
CONCLUSIONS: The modified MRA using SENSE is a cost-effective modality of examination for the demonstration of the whole hepatic vascular system. Combined with MRI, it has the potential as a one-stop imaging modality in the preoperative evaluation in fields such as OLT.

BACKGROUND: Pancreas transplantation is a valid therapy for patients with type 1 diabetes mellitus (IDDM). This study was undertaken to establish a stable pig model of whole pancreatoduodenal allotransplantation of physiologically coincidence for pancreas transplantation.
METHODS: The IDDM pig model was made by resection of the pancreas from the recipient. The portal vein of the donor was end-to-side anastomosed to the anterior mesenteric vein (equal to the superior mesenteric vein) or the portal vein of the recipient. The far end of the celiac axis was anastomosed to each other. With end-to-side fashion, the duodenum of the donor was anastomosed to the jejunum of the recipient.
RESULTS: In 16 transplantations, results were satisfactory. The anatomical results and immune function including the function of the pancreas were similar to those of human being.
CONCLUSION: Of the pancreas of the pig, this study of pancreatic transplantation shows that the novel pig whole pancreatoduodenal allotransplantation model is ideal.

BACKGROUND: Ischemia/reperfusion is the main cause of hepatic damage in liver transplantation. Immediate early genes (IEGs) encode proteins can regulate expression of cellular response genes after injury, and is associated with tissue repair and cell apoptosis. The purpose of this research was to investigate the effects of preconditioning on expression of immediate early genes c-fos and c-jun following hepatic ischemia/reperfusion (IR) and its roles in cellular regeneration and apoptosis.
METHODS: Ninety-six Wistar rats were randomly divided into IR group and hepatic ischemic preconditioning (IPC) group, and each group was further divided into eight sub-groups (n=6). The model of partial liver ischemia/reperfusion was used. The rats were subjected to 60-minute liver ischemia, preceded by 10-minute preconditioning. After 0-, 0.5-, 1-, 2-, 4-, 8-, 12-, 24-hour reperfusion, the serum and liver tissue in each group were collected to detect the level of serum ALT/AST, liver histopathology, expression of c-fos, and c-jun mRNA. Flow cytometer was used to detect Ki67 and Sub-G1 as the quantity indicators of cell regeneration and apoptosis respectively.
RESULTS: Compared with IR group, IPC group showed a significantly lower ALT/AST level in 0.5-hour sub-group to 8-hour sub-group (P<0.05). Ki67 elevated significantly at 0.5, 1, 2 hours, but decreased significantly at 24 hours (P<0.05). Ap index decreased significantly after 1-hour reperfusion（P<0.05). Expressions of c-fos and c-jun mRNA were low, especially c-jun at 0.5, 1 and 2 hours after reperfusion.
CONCLUSION: Ischemic preconditioning can protect liver cells against ischemia/reperfusion injury, and this protective effect may be related to influence transcription levels of c-fos and c-jun.

BACKGROUND: Islet transplantation is considered a potentially curative treatment for diabetic mellitus. The aim of this study was to assess the feasibility of islet transplant through the spleen.
METHODS: Both donor and recipient Wistar rats (BW150±20 g) were provided by the Animal Center of China Medical University, Shenyang, China. Islets were isolated and purified with the modified Minnesota program. 600-800IE graft islets were handpicked and transplanted through the spleen of diabetic recipients. Blood glucose and insulin were evaluated after operation every other day. IVGTT was performed 10 days after transplantation.
RESULTS: 300-400IE islet was procured from one donor rat. Secretion index (SI) of the glucose stress was 5.59±0.62, showing the graft functioning well. The diabetic rats restored normal blood glucose levels of 3.4-5.4 mmol/L (mean 4.8 mmol/L). Their insulin levels were as normal as 8.5-12.2 μIU/ml. The K value of IVGTT in the rats after transplantation was similar to the normal one.
CONCLUSIONS: The islets can be transplanted successfully through the spleen, while avoiding the complications caused by traditional transplantation through the portal vein, such as bleeding and portal vein hypertension. The graft islets loss can be reduced because of less centrifugation and mechanic pressure. In conclusion, transplantation through the spleen is a simple and feasible method.

BACKGROUND: Ischemia/reperfusion (I/R) injury is a major cause of primary graft dysfunction and renders an allograft more immunogenic in orthotopic liver transplantation (OLT). Panax notoginseng saponins (PNS) has been reported to exert protective effects against I/R injury to various organs. The objective of this study is to investigate whether PNS preconditioning protects rat liver grafts from I/R injury via an antiapoptotic pathway.
METHODS: Male Sprague-Dawley rats were used as donors and recipients of orthotopic liver transplantation (OLT) and were divided into PNS preconditioning group(group P) and normal saline control group (group N) randomly according to whether PNS (50 mg/kg) was injected intravenously 1 hour before liver grafts harvesting, and sham group (group S). The animals were separately killed 2, 6 and 24 hours after reperfusion. Plasma samples were collected for test of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Liver tissues were collected to detect histological changes, apoptosis and the expression of TNF-α, Bcl-2 and Caspase-3 mRNA.
RESULTS: The serum levels of ALT and AST and the apoptosis index (AI) of liver tissue in group P were lower than in group N significantly 2, 6 and 24 hours after reperfusion. Compared with group N, the expression of TNF-α and Caspase-3 mRNA was reduced significantly in group P 2 and 6 hours after reperfusion and the expression of Bcl-2 mRNA was enhanced significantly in group P 6 and 24 hours after reperfusion.
CONCLUSIONS: PNS preconditioning protects liver grafts from I/R injury effectively in rat OLT via an antiapoptotic pathway. The antiapoptotic mechanisms of PNS may include inhibiting the expression of TNF-α and Caspase-3 and enhancing the expression of Bcl-2.

BACKGROUND: This study was undertaken to investigate the predictive factors of sustained viral response in roferon-A or pegasys treated chronic hepatitis C patients after logistic regression analysis of the factors that might be associated with the therapeutic effects of interferon (IFN).
METHODS: All patients enrolled into this randomized, open and multi-center controlled trial were divided into two groups randomly and treated with pegasys and roferon-A for 24 weeks, then followed up for another 24 weeks. Before treatment, hepatitis C virus (HCV) genotype was determined, and HCV-RNA in serum was detected before and at the end of treatment and follow-up. HCV-RNA turning negative was considered the major index for evaluating the therapeutic effect. The clinical characteristics including gender, age, infection route of HCV, treatment with IFN, platelet count, AST/ALT ratio and treatment drugs were analyzed by logistic regression.
RESULTS: Intention to treat (ITT) and per-protocol (PP) population groups have 208 and 197 patients respectively. In the PP group, after treatment for 24 weeks, the response rates of female patients aged less than 50 years, infected through non-transfusion, relapsed after IFN treatment, and presented with a AST/ALT ratio≤1, virus load less than 8×105 IU/ml, and non genotype 1 HCV infection, and treated finally with pegasys were higher than those of male patients, aged more than 50 years, infected by transfusion, treated firstly with IFN, presented with a AST/ALT ratio≥1, virus load equal or more than 8×105 IU/ml, and genotype 1 infection, and treated finally with roferon-A. But, at the end of follow-up, the patients with a AST/ALT ratio≥1 and virus load more than 8×105 IU/ml had a higher rate of sustained response than did those with a AST/ALT ratio≤1 and virus load less than 8×105 IU/ml. Logistic regression analysis and control of the promiscuous factors showed that the genotype of HCV was not related to the response rate at the end of treatment (OR 0.604, 95% CI 0.271-1.349， P=0.219), but was the independent predictive factor of virus sustained response to IFN treatment (OR 0.408, 95% CI 0.189-0.881, P=0.023). The form of IFN was significantly related to viral response at the end of treatment (OR 0.105, 95% CI 0.052-0.212, P<0.001), and pegasys were an intensely predictive factor for sustained response(OR 0.255, 95% CI  0.123-0.529, P<0.001).
CONCLUSION: HCV genotype and pegasys are the predictive factors for IFN response to the IFN treatment of patients with chronic hepatitis C.

BACKGROUND: Angiogenesis is known to be essential to the survival, growth, invasion, and metastasis of tumor cells. Vascular endothelial growth factor (VEGF) are an important angiogenic factor regulating tumor angiogenesis, but its significance and tumor pathologic features are unclear in hepatocellular carcinoma (HCC). In the present study, we analyzed expression of tissue VEGF, alteration of microvascular density (MVD) in microvessel angiogenesis, development and metastasis of HCC, and level of serum VEGF in differential diagnosis of benign and malignant liver diseases.
METHODS: Tumor specimens were prospectively collected from HCC patients undergoing resection. Total RNAs were extracted and the expression levels were detected from different parts of HCC tissues. The cellular distributions of VEGF and MVD of liver tumors and their paracancerous and distal cancerous tissues were investigated by streptavidin peroxidase (S-P) immunohistochemistry,  respectively. The VEGF levels of circulating blood and hepatoma tissues were measured by enzyme-linked immunosorbent assay.
RESULTS: The incidence of VEGF expression was 63.9% in HCCs (23/36 cases), 78.3% in non-encapsulated HCCs (18/23), and 90.9% in HCCs with extrahepatic metastasis (10/11), respectively. The VEGF expression was tightly correlated with MVD (P<0.01). The MVD in HCC with metastasis, low differentiation or non-encapsulation was significantly higher than that in HCC with intact capsule, high differentiation, or no metastasis. No significant difference was found between VEGF, MVD, tumor size, and hepatitis virus infection. The level of total RNA in HCC tissues was significantly lower but the VEGF level significantly higher than those in paracancerous or distal cancerous ones (P<0.01). The abnormal expression levels of VEGF in sera of HCC patients were directly correlated with the metastasis and recurrence of tumors.
CONCLUSION: The high expression of VEGF and abnormality of tissue MVD are useful predictors for vascular invasion and metastasis of liver tumors.

BACKGROUND: The highly sensitive technique of reverse transcriptase-polymerase chain reaction(RT-PCR) has been used to detect circulating tumor cells. This study was undertaken to detect circulating hepatocellular carcinoma cells and to evaluate their potential clinical implication in HCC patients.
METHODS: Subjects included 65 patients with primary HCC, 22 patients with hepatitis B and liver cirrhosis, 12 patients with metastatic liver cancer, 11 patients with hepatic hemangioma, and 20 healthy volunteers. AFPmRNA and melanoma antigen gene (MAGE-1) mRNA in peripheral blood samples were tested using nested RT-PCR. 
RESULTS: The positive rates of MAGE-1mRNA and AFPmRNA were 41.5% (27) and 53.8% (35), respectively in the HCC patients. In 64.6%(42) of the 65 patients, there was at least one positive marker. The positive rate of AFPmRNA or MAGE-1mRNA was correlated with portal thrombosis, nodules of tumor, tumor diameter and TNM stage (P<0.05). Samples from 4 (33.3%) of the 12 patients with metastatic liver cancer were MAGE-1mRNA positive, samples from 3(13.6%) of the 22 patients with cirrhosis AFPmRNA positive, and the others were both negative.
CONCLUSIONS: Nested RT-PCR is a sensitive and reliable method for detecting circulating HCC cells. A two-marker RT-PCR assay with a liver-specific AFP marker and a cancer specific MAGE-1 marker may be a promising tool for detecting blood disseminated HCC cells with a better sensitivity and specificity than a single marker RT-PCR.

BACKGROUND: Portal vein tumor thrombosis (PVTT) is a serious complication and a major metastatic way of hepatocellular carcinoma (HCC). But portal vein benign thrombosis (PVBT) always appears in patients with hepatocirrhosis, and PVTT should be differentiated from PVBT. The aim of this study was to probe the value of ultrasound-guided fine needle aspiration biopsy in differential diagnosis of PVTT.
METHODS: Twenty-two HCC patients with portal vein thrombosis and 8 hepatocirrhosis patients with portal vein thrombosis were studied by ultrasound-guided fine needle aspiration biopsy. Twelve portal vein thrombosis filling portal vein embranchment of the 30 portal vein thrombosis patients were examined by 18G automatic biopsy. The positive rates of aspiration biopsy cytology and histology were calculated and compared with those of automatic biopsy.
RESULTS: The positive rates of fine needle aspiration biopsy cytology and histology were 93.3% (28/30) and 90.0% (27/30), respectively. They were not different markedly from that of automatic biopsy 91.7% (11/12). In aspiration biopsy of 22 HCC patients with PVTT, HCC cellular was found in 19 portal vein thrombosis patients (86.4%) by cytology examination and in 18 portal vein thrombosis patients (81.8%) by histology examination. In total, 20 tumor thrombi were detected. The other two were diagnosed as benign thrombosis. No HCC cell and/or tissue was observed in 8 patients with hepatocirrhosis associated with portal vein thrombosis.
CONCLUSIONS: Ultrasound-guided fine needle biopsy in detecting PVTT shows a high positive rate and is of diagnostic value. The positive rate is not apparently different from that of automatic biopsy. Hence the case that fails to be diagnosed by color Doppler flow imaging(CDFI) and pulsed Doppler can be detected early by ultrasound-guided fine needle aspiration biopsy.

BACKGROUND: In recent years, MR techniques have been widely used for displaying hepatic vessels and measuring its hemodynamics, especially in the management of the patients with portal hypertension. The aim of this study was to evaluate the role of magnetic resonance angiography (MRA) in the treatment of patients with portal hypertension.
METHODS: A series of 38 patients with portal hypertension and 12 control patients were enrolled in this study. Three dimensional dynamic contrast-enhanced (3D-DCE) MRA and two dimensional phase-contrast (2D-PC) MR were used for the study of portal venous anatomy and its hemodynamics, which were compared with those of indirect portal-venography (IPVG) and doppler ultrasonography (DUS).
RESULTS: The anatomical imaging of the portal venous system could be displayed clearly in 3D-DCE MRA and the imaging quality of MRA was better than that of IPVG. The hemodynamic information from 2D-PC MR was closely related to that from DUS.
CONCLUSIONS: As a non-invasive technique, MRA can display the anatomy of the portal venous system clearly and measure its hemodynamics exactly. It is very useful and can be applied if necessary in the management of patients with portal hypertension.

BACKGROUND: Signal regulatory protein alpha1 (Sirpα1) is a negative regulatory factor, and inhibits receptor tyrosine kinase-dependent cell proliferating signal. This study was undertaken to observe the effect of signal regulatory proteinα1 (Sirpα1)on gankyrin,cyclin D1,CDK4 and Fas expression in Sk-hep1 mouse hepatoma carcinoma cell line.
METHODS: BOSC 23 packed cells were respectively transfected by means of recombinated retrovirus including pLXSN, pLXSN- Sirpα1 and pLXSN- Sirpα1Δ4Y2 with lipofectin, and  various plasmid virus media(viral titer 2.1×106 CFU/ml) were  collected and infected respectively in 80% confluent Sk-hep1 cells. Transfected Sk-hep1 cells were selectively screened with G418 (1200 μg/ml), and Sk-hep1 cell lines transfected with various plasmids were obtained. The protein expressions of gankyrin, cyclin D1, CDK4 and Fas in various Sk-hep1 lines were determined by Western blotting. Various Sk-hep1 lines were recovered to culture with 10% fetal bovine serum at 12 hours and 24 hours after starving culture with free serum for 72 hours, and cells were collected to determine the percentage of S phase cells of proliferating cycle by flow cytometry.  
RESULTS: Sirpα1 transfection remarkably downregulated gankyrin and cyclin D1 expression. Sirpα1Δ4Y2 downregulation of gankyrin expression was greater than that of Sirpα1 (P<0.05), but no significant effect of Sirpα1 and  Sirpα1Δ4Y2  on CDK4 and Fas protein expression was observed in transfected Sk-hep1 lines (P>0.05). The percentage of S phase cells significantly decreased in Sk-hep1 cells transfected with Sirpα1 and Sirpα1Δ4Y2  plasmids (vs pLXSN Sk-hep1, P<0.05). The percentage of S phase cells in various Sk-hep1 cells increased when recovering to culture with 10% fetal bovine serum at 12 hours, but the percentage of S phase cells in  Sk-hep1 cells transfected with Sirpα1 was the lowest (vs pLXSN and Sirpα1Δ4Y2 Sk-hep1, P<0.05). The percentage of S phase cells in transfected pLSXN Sk-hep1 cells was the largest (vs Sirpα1 and Sirpα1Δ4Y2 Sk-hep1, P<0.05). There was no significant difference between the transfected Sirpα1 Sk-hep1 cells and Sirpα1Δ4Y2 Sk-hep1 cells (P>0.05).
CONCLUSIONS: Sirpα1 decreases gankyrin and cyclin D1 expression, and inhibits proliferation of liver carcinoma cells. It may be one of the forms for an Sirpα1 negative regulation of carcinogenesis and development of hepatocellular carcinoma.

BACKGROUND: The demand for the clinical use of hepatocytes is increasing. The aim of this study was to develop a method for procurement of high qualitative pig hepatocytes and to evaluate the state of freshly isolated and cultured hepatocytes.
METHODS: The domestic extracorporeal circulating perfusion apparatus was used to isolate and harvest swine hepatocytes by the two-step perfusion method with EDTA and collagenase. The  viability, function and morphology of the freshly isolated and cultured cells were evaluated and observed by the trypan blue exclusion test, biochemical measurements, phase contrast microscopy and transmission electron micrography (TEM).
RESULTS: The total yield of isolated hepatocytes reached to 1.5(±0.4)×1010 per liver with a viability of 92(±5)%, and the purity of hepatocytes reached to 98%. Immediately after isolation, phase-contrast microscope and TEM showed that undamaged hepatocytes appeared bright, translucent and spherical in shape, with a characteristic well-contrasted border. After 24 hours, the concentrations of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), albumin (ALB), creatinine (Cr) and blood urea nitrogen (BUN) in the fluid of culture were declined significantly.
CONCLUSION: This method of procuring swine hepatocytes could get high quality cells with active metabolic function.

BACKGROUND: Massive hemobilia is a rare but potentially life-threatening cause of upper gastrointestinal hemorrhage. Transarterial embolization is considered the first line of intervention to stop the bleeding for most causes of hemobilia. This study was conducted to evaluate selective hepatic angiography and embolization in the diagnosis and treatment of patients with massive hemobilia. 
METHODS: The clinical data of 16 patients with massive hemobilia were analyzed retrospectively. These patients underwent emergency celiac and selective right or left hepatic artery angiography and treated by embolization using Gelfoam particles and/or coils.
RESULTS: Hepatic artery angiography revealed hepatic artery pseudoaneurysms in 6 patients, cystic artery pseudoaneurysms in 2, diffuse hemorrhage of hepatic artery branches in 5, and right hepatic artery-bile duct fistulae in 3. The patients were diagnosed rapidly by angiography and treated successfully by embolization of the hepatic artery branch proximal to the bleeding point, and hemorrhage was stopped immediately. Two patients were embolized the second time for rebleeding. Neither recurrence of bleeding nor serious complication was found during the follow-up for 3 months to 2 years. The other 2 patients whose hemorrhage failed to be controlled died several days later.
CONCLUSION: Being safe, reliable and minimally invasive, selective hepatic artery angiography and embolization are effective in the diagnosis and treatment of massive hemobilia.

BACKGROUND: Overexpression of human tissue inhibitors of metalloproteinase-1 (TIMP-1) may play an antitumor role in inhibiting hepatocellular carcinoma (HCC) growth and progression. The aim of the study was to construct a recombinant adenovirus vector carrying hTIMP-1 cDNA for liver gene therapy and observe its expression in vitro.
METHODS: The full-length cDNA of hTIMP-1 was positively cloned into the adenoviral shuttle vector pAdTrack-CMV, and then cotransformed into competent BJ5183 cells with the adenoviral backbone plasmid pAdEasy-1. Thus, a recombinant adenovirus AdhTIMP-1 containing full-length cDNA of hTIMP-1 was generated by homologous recombination in E. coli. AdhTIMP-1 was then packaged and amplified in adenoviral packaging cells, or human embryonic kidney 293 cells. The viral titer was checked by green fluorescent protein (GFP), and the expression of hTIMP-1 in vitro was detected by the techniques of Western blot and RT-PCR.
RESULTS: The recombinant adenovirus vector carrying hTIMP-1 was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. The transcription of TIMP-1 mRNA in 293 cells was checked by RT-PCR and TIMP-1 protein could be detected in the culture by Western blot analysis.
CONCLUSION: The successful construction of recombinant adenoviral vector carrying human TIMP-1 and the effective expression in vitro has laid a foundation for further study of its antitumor function and may pave the way for future application in liver gene therapy.

BACKGROUND: Alcoholic liver disease (ALD) is a serious and potentially fetal consequence of alcohol use. The diagnosis of ALD is based on alcohol consumption, physical signs and symptoms, and laboratory tests. The aim of the present study was to assess the reliability of carbohydrate-deficient transferrin (CDT) in the diagnosis of ALD.
METHODS: According to the diagnostic criteria for ALD by the Chinese Medical Association in 1995, 76 patients with ALD, 55 patients with alcoholism, 32 patients with nonalcoholic liver disease (NALD), and 27 healthy subjects (controls) were studied. Serum CDT was assayed by isoelectric focusing immunofixation and Comassie blue staining. The levels of alamine aminotransferase (ALT), aspartate aminotransferase (AST), and gamma glutamyltransferase (GGT) were also examined.
RESULTS: The positive rate of CDT in the patients with ALD was 93.4%(71/76), which was higher than that in those with alcoholism (52.7%, 29/55, P<0.001), in those with NALD(9.4%, 3/32, P<0.001), and in healthy controls, respectively. The sensitivity and specificity of CDT for ALD was 93.4% and 71.9%, respectively.
CONCLUSION: CDT may help diagnose alcoholic liver disease.

BACKGROUND: The effect of tacrolimus (FK506) and 5-fluorouracil (5-FU) on hepatocellular carcinoma remains elusive. The aim of this study was to assess the effect of tacrolimus on the proliferation, and apoptosis in liver cancer cell line of SMMC-7721 and its sensitivity to fluorouracil (5-FU).
METHODS: The liver cancer cell line of SMMC-7721 was cultured in vitro, and the MTT assay was used to examine the antiproliferative effect of FK506. Flow cytometry (FCM) was used to examine the effect of 5-FU alone or in combination with FK506  on the apoptosis and cell cycle of SMMC-7721 cells.
RESULTS: FK506 produced concentration-dependent antiproliferative effect on SMMC-7721 cells at all experimental concentrations（P<0.05）, but no effect on induction of apoptosis. 5-FU induced apoptosis in a concentration-dependent manner, whereas the percentage of G0/G1-phase cells and proliferation index (PI) were increased with the increased concentration of 5-FU. Pretreatment with FK506 for 2 hours enhanced the effect of 5-FU on the induction of apoptosis.
CONCLUSIONS: FK506 inhibits the growth of SMMC-7721 cells and enhances their sensitivity to 5-FU. This may be associated with the synergic effect of FK506 and 5-FU in inducing apoptosis and G0/G1-phase stasis.

BACKGROUND: Hepatic ischemia/reperfusion injury may induce intestinal microflora imbalance. Salvia miltiorrhiza is effective in promoting blood circulation and counteracting peroxidation in tissues. The aim of the present study was to determine the effects of Salvia miltiorrhiza on intestinal microflora, endotoxemia, and bacterial translocation in rats with hepatic I/R injury.
METHODS: Sprague-Dawley rats in specific pathogen free grade were divided into 3 groups: group I (n=6) for sham operation; groups II (n=10) and III (n=7) for liver ischemia for 20 minutes and reperfusion for 22 hours. Group III was also pretreated with 4 ml/day of Salvia miltiorrhiza solution (250 mg/kg) by daily gavage for 7 days. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA) and superoxide dismutase(SOD) in liver tissues, serum endotoxin, intestinal bacterial counts, intestinal mucosal histology and bacterial translocation were studied.
RESULTS: The levels of ALT, AST, plasma endotoxin and MDA in liver tissues were decreased more markedly in group III (57.57±18.08 U/L, 147.57±40.84 U/L, 0.42±0.144 EU/ml and 0.52±0.19 nmol/mg-prot respectively) in group II (122.8±80.12 U/L, 295.9±216.92 U/L, 0.80±0.262 EU/ml and 0.72±0.12 nmol/mg-prot; P<0.05-0.01 respectively). Liver SOD activity was increased more significantly in group III (318.47±64.62 U/mg-prot) than in group II (240.76±63.67 U/mg-prot, P<0.05). The counts of Bifidobacteria and Bacteroides increased more significantly in group III than in group II, but were similar to those in group I. Bacterial translocation to the kidney in group II was 50%(5/10), whereas no bacterial translocation to the kidney occurred in the other two groups (P<0.01). Ileal mucosal structure was markedly ameliorated in group III as compared with group II.
CONCLUSIONS: Salviae miltiorrhiza could partially restore intestinal microflora balance, improve intestinal mucosal integrity, and reduce bacterial translocation and plasma endotoxin in rats with hepatic ischemia/reperfusion injury.

BACKGROUND: Playing a central role in hepatic fibrosis, hepatic stellate cell (HSC) has made itself the major target of research. The limited supply of HSC, however, can not meet the ever growing need of experiment. Establishment and identification of novel immortalized HSC line thus may be urgently required.
METHODS: Primary HSCs were isolated from a normal adult male Sprague-Dawley rat by in situ perfusion with collagenase IV and pronase E, and then were purified by single-step density gradient centrifugation with nycodenz. Once they reached total activation in culture, a new immortalized myofibroblast-like HSC line was established through cellular cloning. Its characteristics were identified by means of immunocytochemical staining, light microscopy, transmission electron microscopy, and growth curve analysis.
RESULTS: The novel HSC line, termed HSC-PQ, had a doubling time of about 75 hours in the Dulbecco’s modified Eagle medium (DMEM) containing 20% fetal bovine serum. Most of the main morphological characteristics of the differentiated primary HSC could be detected in HSC-PQ cell, while functional features of activated HSC such as α-smooth muscle actin, desmin, collagen type I, collagen type III, fibronectin, laminin and other extracellular matrix proteins could also be found in it except for collagen type IV. In contrast, fat droplets and autofluorescence of vitamin A disappeared in the HSC-PQ line. This cell line had been maintained in culture for over 30 passages and more than 1 year with little alternation in biological characteristics.
CONCLUSION: A new rat HSC line (HSC-PQ) has been successfully established. It consistently retains the characteristics of activated primary HSC, and has proved to be immortalized.

BACKGROUND: Percutaneous ethanol injection has been widely used as a non-surgical therapy for liver cancer, but it has some shortcomings such as local diffusion and unequal permeation. This study was designed to observe the volume, controllability and completeness of necrosis after injection of low concentration sodium hydroxide in the normal liver parenchyma so as to assess its possibility in treatment of liver cancer instead of ethanol.
METHODS: Twenty-seven New Zealand rabbits were divided randomly into 9 groups (Aa, Ab, Ac, Ba, Bb, Bc, Ca, Cb, and Cc) by a 3×3(three-by-three) factorial design, each consisting of 3 rabbits. Group A was given sodium hydroxide solution at a concentration of 5%, while B at 2.5% and C at 1% in liver parenchyma. Each group received three doses of the solution: a (0.2 ml), b (0.5 ml) and c (1.0 ml). Then another 3 rabbits as side-effect group were dropped with sodium hydroxide solution in their liver lobe space. Liver and renal function changes in all the rabbits were compared after injection with pre-injection.
RESULTS: All the lesions were localized. At the concentration of 2.5% and 5%, the lesion volume increased with the dose increased from 0.2 ml to 1.0 ml (P<0.05). No significant differences were found in the lesion volume of the groups receiving the same dose but different concentration. Changes in liver and renal function were not significant 7 days after injection, compared with those before injection.
CONCLUSIONS: 2.5% and 5% sodium hydroxide solution could control local complete necrosis in normal liver. With regard to safety, 2.5% alkali solution is considered promising as a new agent for intratumoral injection therapy instead of ethanol.

BACKGROUND: There have been lots of important discoveries in the research on embryonic stem (ES) cells; but few studies have been involving in the differentiation and differentiation ratio of hepatic stem cells. To provide a new source of hepatocytes in hepatocytes transplantation and liver tissue engineering, we investigated the differentiation and differentiation ratio of hepatocytes in the ES cells differentiating system in a certain length of time.
METHODS: BALB/c ES cells from mice were cultured for differentiation in medium without leukemia inhibitory factor (LIF). During the period of differentiation of ES cells, dual immunofluorescence was performed to analyze the expression of hepatic proteins in cytoplasm such as α-fetoprotein (AFP) and albumin (ALB). The count of relative hepatocyte-like cells was analyzed by flow cytometry (FCM) and the hepatic differentiation ratio was obtained.
RESULTS: The expression of AFP was first determined at day 7 and that of ALB at day 13. The expressions of AFP and ALB became stronger with the extention of the differentiation time. The hepatic differentiation ratio at day 13 and 21 was 5.5% and 10.4%, respectively.
CONCLUSION: ES cells could differentiate into hepatocytes, which should be a new source of hepatocytes in transplantation of hepatocytes and liver engineering.

BACKGROUND: The incidence of gallstone is higher in patients with diabetes mellitus than in general population because of hypomotility and lowered emptying function. The aim of this study was to investigate the gene expression of cholecystokinin-A (CCK-A) receptor in patients with gallstones and diabetes mellitus and its correlation with the hypomotility of the gallbladder.
METHODS: Smooth muscle was taken from the gallbladder after cholecystectomy and total RNA was extracted from it by Trizol reagent to compose cDNA. CCK-A receptor was enlarged by reverse transcription-polymerase chain reaction (RT-PCR) and identified by electrophoresis. Based on the analysis of images, the relative expression of the CCK-A receptor was obtained after comparison of the CCK-A receptor and β-action. The emptying function of the gallbladder was measured with B ultrasound before operation.
RESULTS: Gallbladder ejetion fraction (GBEF2)(%) (24.9±12.7) decreased more significantly in patients with gallstones and diabetes mellitus than in those with simple gallstone (61.5±8.5)(P<0.01). In the two groups, a fraction of 540bp of CCK-A receptor could be testified respectively, but the relative expression was different. Compared to the expression of β-action, it was 0.400±0.068 in patients with gallstones and diabetes mellitus, but 0.622±0.070 in those with gallstone only (F=7.169,  P<0.01). The result was consistent with the decreased contractility.
CONCLUSIONS: The gene expression of CCK-A receptor on the smooth muscle of the gallbladder may decrease more significantly in patients with gallstone and diabetes mellitus than in those with gallstone only; this result is consistent with the decreased motility. Thus the lowered expression of CCK-A receptor is probably the cause of hypomotility in patients with gallstones and diabetes mellitus.

BACKGROUND: There is a controversy about the risk of  injury to the branch of the middle hepatic vein during laparoscopic cholecystectomy. This study was conducted to further investigate the relationship between the gallbladder bed and the branch of the middle hepatic vein.
METHODS: Color Doppler ultrasound was used to examine the anatomical relationship between the gallbladder bed and the branches of the middle hepatic vein in 143 healthy volunteers.
RESULTS: Not all the middle hepatic vein extended close to the gallbladder bed, the branches and gallbladder beds in 23 subjects were not in the same plane during ultrasound scanning. In 21 of the 143 subjects the branch of the middle hepatic vein was completely adherent to the gallbladder bed with a diameter ranging from 1.2 mm to 3.6 mm. In 10 subjects the branches of the middle hepatic vein traversed approximately 1.0 mm from the gallbladder bed with a diameter ranging from 1.6 mm to 3.0 mm.
CONCLUSIONS: In most subjects the branch of the middle hepatic vein and the gallbladder bed are well separated. Only patients with large branches of the middle hepatic vein close to the gallbladder bed are at risk of hemorrhage during laparoscopic cholecystectomy.

BACKGROUND: Somatostatin, a neuropeptide and hormone, exists in the biliary tract of several species. The effects of somatostatin and its analogues on the sphincter of Oddi motility have been controversial. The aim of this study was to observe the action of stilamin and sandostatin on the sphincter of Oddi via choledochofiberscope manometry.
METHODS: Twenty patients who had had “T” duct after cholecystectomy and choledochotomy were divided into 2 groups randomly: stilamin and sandostatin. They were subjected to manometry via a choledochofiberscope through the‘T’duct tract. The following data recorded included duodenal pressure (DP), sphincter of Oddi basal pressure (SOBP), sphincter of Oddi contractive amplitude (SOCA), frequency of the sphincter of Oddi (SOF), duration of the sphincter of Oddi, and the common bile duct pressure (CBDP).
RESULTS: After intravenous administration of stilamin at a dose of 250 μg/h, the mean SOCA increased from 89.18(26.50) to 128.57(54.21) mmHg (P<0.05). After the administration of stilamin at a dose of 500 μg/h the mean SOCA declined to 92.18(42.81) mmHg (P<0.05), and mean SOBP declined from 17.63(13.36) to 8.16(4.01) mmHg (P<0.05). Although SOF had declined from 9.25(2.45) to 7.46(1.52) n/min, it was not significantly influenced. After intravenous administration of sandostatin at a dose of 100 μg, the mean CBDP increased obviously.
CONCLUSIONS: Intravenous administration of stilamin at a dose of 250 μg/h stimulates the motility of the sphincter of Oddi whereas the injection of stilamin at a dose of 500 μg/h inhibits its motility. Intravenous injection of sandostatin of 100 μg has no effect on the sphincter of Oddi.

Metastatic clear cell carcinomas are commonly seen in the kidney and lung. Clear cell variant of hepatocellular carcinoma is an uncommon tumour. Diagnosis is usually made by correlation of histopathology, tumour markers and immunohistochemistry, especially HepPar 1. In this unusual case of metastatic clear cell carcinoma presenting as Sister Mary Joseph’s nodule, no primary evidence was observed radiologically in the liver, but the level of alfa fetoprotein was markedly elevated. Metastatic clear cell carcinoma of the liver with an occult hepatic primary was diagnosed by immunohistochemical profile of the tumour.

A 65-year-old man presented with bone pains and anemia. Skull X-ray revealed multiple osteolytic lesions. The patient was evaluated for multiple myeloma but detailed workup revealed the diagnosis of primary hepatocellular carcinoma (HCC) with osteolytic bone metastases. Thus, bone metastases due to HCC, although rare, should be considered in patients presenting with bone pains due to osteolytic lesions.

BACKGROUND: Congenital choledochal cyst is a rare kind of bile duct deformity, resulting from cystic or shuttle-like dilation of part of the choledochal duct congenitally. We present a 20-year-old girl with a congenital choledochal cyst complicated by acute pancreatitis.
METHOD: The clinical data of the woman with a congenital choledochal cyst concurrent with acute pancreatitis were retrospectively analyzed.
RESULT: The congenital choledochal cyst of the woman was type IV complicated by acute pancreatitis.
CONCLUSIONS: The diagnosis of congenital choledochal cyst mainly depends on CT, MRCP, and ERCP. Total excision of the choledochal cyst with Roux-en-Y hepaticojejunostomy is recommended as the treatment. For patients with type V cysts with frequently recurrent cholangitis resulting biliary liver cirrhosis, liver transplantation should be considered.

BACKGROUND: Despite its rare incidence, few cases of left-side gallbladder have been already published.
METHODS: We reported herein  the case of a 29-year-old man with a left liver tumor in whom left lateral bisegmentectomy was mandatory. It represents the first description of a sinistroposition of both gallbladder and common bile duct.
RESULTS: Surgical exploration revealed a left-side gallbladder, located under the left lobe of the liver. During hepatic parenchyma dissection at the left side of the round ligament and the Rex recessus, the common bile duct was injured. Complete separation of hepatic pedicle structures showed that the upper biliary convergence passed on the left side of the Rex recessus before reaching the hepatoduodenal ligament.
CONCLUSION: Only careful dissection of the hepatoduodenal ligament up to Rex recessus level prior to liver parenchyma resection could avoid biliary tract injury during left lobectomy.