BACKGROUND: Gene therapy as part of modern molecular medicine holds great promise for the treatment of hepatocellular carcinoma (HCC) and has the potential to bring a revolutionary era to cancer treatment. For the past decade various viral and non-viral vectors have been engineered for improved liver gene therapy.
DATA RESOURCES: An English-language literature search using MEDLINE (2004), Index Medicus (2004) and bibliographic reviews of books and review articles. Liver-directed gene transfer vectors and their history and recent clinical applications.
RESULTS: The ultimate goal of liver-directed gene therapy for HCC is the stable expression of a therapeutic transgene in a significant proportion of hepatocytes. The design of a vector system providing efficient and stable gene engraftment and expression in human hepatocytes is still a challenging issue. The advantages and disadvantages of the genetically engineered vector of viral or non-viral origin are discussed with respect to their essential relevance.
CONCLUSION: Liver gene therapy has a long way to go and efficient and innocuous liver-directed gene transfer vectors are therefore urgently required.

BACKGROUND: Interferon-alfa has been used in the treatment of chronic hepatitis B for more than 20 years and has its own advantages including a definite course of therapy, no production of drug-resistant variants, and sustained efficacy. This review was to understand the role of interferon-alfa therapy in chronic hepatitis B.
DATA RESOURCES: An English-language literature search using Medscape and MEDLINE was performed and a total of 48 articles on the treatment of chronic hepatitis with interferon-alfa or pegylated interferon-alfa were selected.
RESULTS: Interferon-alfa therapy was associated with a higher HBV DNA inhibition rate and HBeAg loss rate compared with controls, and it may have long-term beneficial effects in terms of HBV clearance, reduction of hepatocellular carcinoma, and prolongation of survival. Pegylated interferon-alfa was more effective than conventional interferon-alfa in the treatment of chronic hepatitis B as well as chronic hepatitis C, and was also associated with greater efficacy than conventional interferon in difficult-to-treat disease.
CONCLUSIONS: Interferon-alfa is still regarded as one of the first-line drugs for the treatment of chronic hepatitis B. Pegylated interferon is a more promising therapy than conventional interferon-alfa, especially in patients with refractory chronic hepatitis B.

BACKGROUND: Orthotopic liver transplantation (OLT) has been the valuable treatment of choice for patients with hepatocellular carcinoma (HCC). As the number of patients with portal vein tumor thrombi (PVTT) increases, most transplant centers become suspicious of the exact effect of the operation which has been accepted as a radical method. This study was designed to evaluate the clinical effects of OLT for patients with HCC associated with PVTT.
METHODS: The follow-up of 24 patients with HCC complicated by PVTT who had received OLT (transplant group) from January 1999 to March 2003 was compared to that of 27 patients undergoing routine hepatic resection (resection group) and 59 patients without surgical treatment (non-surgical group).
RESULTS: The perioperative mortality was zero for the transplant group. The 6-month, 1-year, and 2-year overall survival rates were 66.7%, 29.5% and 23.6% for the transplant group, 33.3%, 22.2% and 14.8% for the resection group (P=0.0335), and 42.1%, 24.4% and 4.1% for the non-surgical group, respectively (P=0.0316). The tumor free survival rates of recipients at 6-month, 1-year, and 2-year were 51.5%, 23.2% and zero, respectively. During the period of follow-up, the overall post-transplant intrahepatic recurrence or extrahepatic metastasis rate was 66.7% for the transplant group.
CONCLUSION: OLT is an effective but palliative treatment modality for patients with HCC associated with PVTT followed by a prolonged survival but a poor tumor free survival rate.

BACKGROUND: Hepatitis B virus reinfection is an important problem after liver transplantation. The aim of this study was to discuss the prevention of hepatitis B virus reinfection following orthotopic liver transplantation.
METHODS: Sixty-eight cases of chronic fulminant hepatitis B, end-stage liver cirrhosis, and liver carcinoma complicated with HBV cirrhosis were given anti-viral drugs before and after transplantation to prevent hepatitis B virus reinfection. Lamivudine was administered in 2 patients, lamivudine+hepatitis B immunoglobulin (HBIG) in 63, and adefovir+HBIG in 3. The measurement of serum HBV, HBV DNA, liver biopsy immunohistochemistry and clinical study were performed.
RESULTS: In 1 of the 2 patients who developed reinfection after lamivudine administration, serum HBsAg, HBeAb, HBcAb, HBV DNA were positive and liver biopsy immunohistochemistry showed HBsAg phenotype. In 2 of 63 patients who  developed reinfection after use of lamivudine+HBIG, serum HBsAg, HBeAb, HBcAb were positive and liver biopsy immunohistochemistry showed HBsAg phenotype. Serum HBV DNA was positive in one of them. Three patients developed no reinfection with HBV after use of adefovir.
CONCLUSIONS: Orthotopic liver transplantation is effective in the treatment of HBV-infected diseases. Lamivudine+HBIG or adefovir+HBIG could effectively prevent hepatitis B virus reinfection.

BACKGROUND: Post-transplantation pancreatitis and graft thrombosis are two major complications of pancreas transplantation that contribute to morbidity,  mortality, and graft loss. Nitric oxide(NO) is a potent vasodilator agent formed when L-arginine(L-Arg) is converted to L-citrulline by the action of NO synthase (NOS), and plays a major role in microcirculatory changes. We therefore investigated the effect of L-Arg on reperfusion injury following pancreaticoduodenal transplantation in rats.
METHODS: The homologous male Wistar rat model of heterotopic total pancreaticoduodenal transplantation was used. The L-Arg-treated rats received the intravenous injection of L-Arg 5 minutes before and after reperfusion at a dose of 200 mg/kg while the N-Nitro-L-arginine methyl ester (L-NAME)-treated rats at a dose of 10 mg/kg. The
amount of NO in the pancreas graft was measured. Serum concentration of cytokine-induced neutrophil chemoattractant (CINC) was determined by enzyme-linked immunosorbant assay, the expression of CINC mRNA was detected by Northern blot assay in the pancreas graft, and the activity of myeloperoxidase (MPO) was measured. Histological examination was performed. 
RESULTS: The amount of NO was higher in the L- Arg group than in the control group, while it was lower in the L-NAME group than in the control group (P<0.05). The peak of serum CINC concentration occurred 3 hours after reperfusion with the difference among the groups being significant. The expression peak of CINC mRNA in the pancreas graft occurred 3 hours after reperfusion. The expression level in the L-Arg group (7.66±1.53   μg/L) was lower than in the control group (26.31±2.01 μg/L), while in the L-NAME group (34.18±3.12 μg/L) it was higher than that in the control group (P<0.05). The activity of MPO in the L-Arg group was obviously decreasd as compared with in the other groups. The pancreas inflammation was ameliorated when L-Arg was administered, whereas the pancreas damage was aggravated when L-NAME was administered.
CONCLUSIONS: L-Arg can increase the amount of NO and inhibit the elevation of CINC, the CINC mRNA expression and early neutrophil accumulation in the pancreas. NO has protective effects on ischemia/reperfusion injury in pancreaticoduodenal transplantation.

BACKGROUND: The highly specific vascular endothelial growth factor (VEGF) induces the growth of vascular endothelial cell. This study was to construct the eukaryotic expression plasmid of vascular endothelial growth factor165 (VEGF165) and observe its expression in vascular smooth muscles (VSMCs).
METHODS: The primers were designed and synthesized according to the gene sequences of human VEGF165. The VEGF165 gene was obtained from umbilic artery tissue by the method of RT-PCR, then it was cloned to eukaryotic expression plasmid pBudCE4.1 by recombination strategy. The eukaryotic expression plasmid named pBudCE4.1/VEGF165 was identified by restriction enzyme digestion, and was sequenced. The pBudCE4.1/VEGF165 was transfected into VSMCs by using lipofection. The VEGF165 expression of mRNA and protein was detected by RT-PCR and Western blot respectively.
RESULTS: VEGF165 was shown about 576bp by RT-PCR. Sequencing revealed the amplified VEGF165 gene was identical with that in the GeneBank. Restrictive enzyme (Hind Ⅲ, Bam HI) digestion analysis showed that recombinant expression plasmid pBudCE4.1/tVEGF165 had been constructed successfully. The expression of VEGF165 at mRNA and protein levels in the transformed VSMCs had been demonstrated by RT-PCR and Western blot.
CONCLUSIONS: The recombinant eukaryotic expression plasmid pBudCE4.1/VEGF165 has been successfully constructed and expressed in transformed VSMCs. The present study has laid a  foundation for VEGF165 gene therapy of vascular stenosis in the transplant organ.

BACKGROUND: Heme oxygenase-1 (HO-1)-mediated cytoprotection may inhibit or postpone the formation of graft arteriosclerosis. The aim of this study was to construct eukaryotic expression vector of HO-1 gene and express recombinant HO-1 in human endothelial cells of the umbilical vein to provide an important clue for chronic rejection study.
METHODS: HO-1 gene was amplified from rat spleen with reverse transcription-polymerase chain reaction, and then cloned into eukaryotic expression vector pcDNA3 by means of recombinant gene technology. The recombinant plasmid pcDNA3-HO-1 was transfected into endothelial cells, and recombinant HO-1 was expressed in the endothelial cells under G418 selection. The expressed products were detected using indirect fluorescent staining and Western blot analysis.
RESULTS: Restriction analysis indicated that the HO-1 gene was successfully inserted into the expression vector, and DNA sequencing verified that the reading frame of the recombinant vector was correct. Recombinant HO-1 was successfully expressed in endothelial cells and its molecular weight was about 32 kD.
CONCLUSION: The successful construction of eukaryotic expression vector containing the HO-1 gene and the effective expression of recombinant HO-1 in endothelial cells has laid a foundation for further study on its biological functions.

BACKGROUND: Liver cancer is one of the most common diseases around the world. The aim of this study was to verify the effect of magnetic field application on target distribution of nanoparticles in transplanted rat liver cancer model and to find out a new method for the treatment of malignant liver tumor.
METHODS: Seven days after the establishment of the model, the abdomen of the rat was exposed through a midline abdominal incision. A cannula was inserted into the gastro-duodenal artery. In the experimental group (12 rats), the tumor tissue was exposed to the magnetic field for 30 minutes.Magnetic albumin nanoparticles containing adriamycin or at an equal dose of free adriamycin (0.5 mg/kg) were injected into the hepatic artery. After the magnetic field was removed, the rat was immediately sacrificed. An equal dose of nanoparticles in absence of the magnetic field served as control (12 rats). Tissues of tumor, nontargeted sites of the liver, heart, kidney, lung, spleen, stomach and small intestine were analyzed for γ-counts and examined histologically.
RESULTS: In the experimental group, the radioactivity of tumor tissue was 8.7 times that of liver tissue. In the control group, the radioactivity of tumor tissue was 2.8 times that of normal liver tissue. The radioactivity of the lung was reduced more significantly in the experimental group than in the control group. No significant difference in the kidney, heart, spleen, small intestine and stomach was observed between the experimental group and control group. And over 80%of the injected nanoparticles distributed in the liver.
CONCLUSIONS: In the presence of magnetic field, magnetic albumin nanoparticles may accumulate in tumor tissues, of which the radioactivity can increase to 8.7 times that of normal liver. Even if the magnetic field is not applied, magnetic albumin nanoparticles in tumor tissues still increase to 2.8 times that of normal liver tissues. These findings indicate that normal organs in the presence of magnetic field are less exposed to chemotherapeutic drugs.

BACKGROUND: Some factors have been reported to be sassociated with a greater likelihood of sustained viral response (SVR) in the interferon (IFN) treatment of chronic hepatitis C. The factors include HCV genotype, HCV RNA level in serum, state of liver disease, baseline body weight, age, sex, and race. The aim of this trial was to investigate the influence of HCV genotype on the IFN treatment of patients with chronic hepatitis C.
METHODS: The genotypes of HCV virus were determined in the patients with chronic hepatitis C from several hospitals of China enrolled into the randomized, opened and controlled trial of Peg-IFN alpha-2a (pegasys) treatment, controlled with IFN-α-2a (roferon-A). The serum ALT levels and HCV RNA concentrations of the patients were detected before and at the end of treatment and during the follow-up. The influence of HCV genotype on the IFN treatment of patients with chronic hepatitis C was analyzed in intention-to-treat (ITT) population.
RESULTS: The HCV genotypes of 202 patients were determined. Of these patients, 158(78.22%) were infected with genotype 1 HCV and 44(21.78%) with genotype non-1. The viral response at the end of treatment (ETVR) and sustained viral response (SVR) rates were 53.80% and 25.32% respectively in patients with genotype 1 HCV, but they were 61.36% and 43.18% in patients with genotype non-1. The difference of SVR between patients with genotype 1 HCV and those with genotype non-1 was significant (P=0.021). After being grouped by the used drugs, the ETVR rates of patients infected with genotype 1 and non-1 HCV were 76.83% and 80.95% in the patients treated with pegasys (P=0.686); but their SVR rates were 35.37% and 66.67% (P=0.01). The viral relapse rate of genotype 1 HCV (55.56%) was significantly higher than that of genotype non-1 HCV (23.53%) (P=0.02). In roferon-A group, the ETVR and SVR rates of patients with genotype 1 HCV were 28.95% and 14.47% respectively, which were lower but not more significant than those of patients with genotype non-1 HCV (43.48% and 21.74%). Moreover, the viral relapse rate of genotype 1 HCV (72.73%) was higher but not more significant than that of genotype non-1 HCV (50.00%) (P=0.21).
CONCLUSION: HCV genotype could affect the efficacies, mainly sustained responses, of IFN treatment in patients with chronic hepatitis C, and the effects of IFN are related to  drugs and therapeutic course.

BACKGROUND: Currently, the managment of non-alcoholic fatty liver disease (NAFLD) is less than certain. Some choleretic might be of potential benefit and deserve further evaluation. This multicenter clinical trial was designed to evaluate the efficacy and safety of Chinese herbal medicine Danning Pian (composed of rhubarb, grant knotweed, dried green orange peel and dried old orange peel) in the short-term treatment of patients with NAFLD.
METHODS: The efficacy and safety of Danning Pian in the short-term treatment of NAFLD were investigated in 232 patients by a multicenter clinical trial during the period of July 1999 to February 2000. The patients consisting of 189 males and 43 females with an average age of 46.1±8.7 years were given 3-5 tablets of Danning Pian orally thrice daily for 3 months in addition to the other comprehensive therapy. The effects of Danning Pian on NAFLD were evaluated by the improvement of clinical symptoms, blood lipids, hepatic enzymes and liver ultrasonographic features. The drug safety was monitored by physical examinations, vital signs, and laboratory tests in addition to the assessment of the adverse events.
RESULTS: All the enrolled patients completed the study except one whose serum ALT level was moderately increased during the therapy with Danning Pian. The effective rate of Danning Pian for the improvement of clinical symptoms, serum ALT levels, blood lipid and fatty liver was 85.8%, 78.2%, 39.6% and 34.0% respectively after the therapy for 3 months. However, the reduction of excessive body weight and waistline did not reach the significant level on the whole after the therapy. The general mild adverse events included diarrhea, skin rash and mild to moderate elevation of serum ALT level. The incidence of adverse reaction was 15.1%.
CONCLUSION: The data of this trial indicate that Danning Pian is effective and safe, generally well-tolerated without severe adverse events, in the treatment of patients with NAFLD over a 3-month period.

BACKGROUND: The mortality rate of heavy type hepatitis is high. No special treatment is available except general treatment. This multicenter clinical study was designed to observe the safety and efficacy of promoting hepatic growth factor (PHGF) in the treatment of heavy type hepatitis and severe chronic hepatitis.
METHODS: 347 patients with heavy type hepatitis and 324 with severe chronic hepatitis were subjected to administration of 120 μg of PHGF per day for 4 weeks on the basis of general treatment. Those who were being effectively treated would last additional 2 to 4 weeks. Blood routine, urine routine, blood urea nitrogen (BUN), blood creatinine (Cr), blood ammonia, alpha fetoprotein (AFP), electrolyte, alanine transaminase (ALT), aspartate transaminase (AST), serum total bilirubin (TBIL), serum direct bilirubin (DBIL), prothrombin time activity (PTA), total protein (TP) and albumin (ALB) were detected in the patients before treatment, 2 weeks after treatment, and at the end of the treatment. Any side-effect would be recorded.
RESULTS: In the patients with severe chronic hepatitis, the total effective rate of the treatment was 88.9%. The levels of ALT, AST and TBIL decreased significantly (P<0.001), whereas those of PTA and ALB increased significantly (P<0.001), and the level of AFP increased slightly. In patients with heavy type hepatitis, the total effective rate of this treatment was 78.4%, and patients at different stage showed different results. The total effective rates of patients with early, medium and terminal stage heavy type hepatitis were 89.9%, 84.8% and 27.5%, respectively. No severe side-effect was shown.
CONCLUSION: PHGF is effective and safe in the treatment of patients with heavy type hepatitis and severe chronic hepatitis. But it should be administered early in patients with heavy type hepatitis so as to get better curative effects.

BACKGROUND: Hypoxia up-regulates vascular endothelial growth factor (VEGF) and stimulates the growth of hepatocellular carcinoma (HCC) cells. This study was designed to investigate the association between changes in plasma VEGF levels after transcatheter arterial chemoembolization (TACE) and HCC  progression, especially in relation to metastasis.
METHODS: Plasma VEGF levels were measured by quatitative sandwich enzyme-linked immunosorbent assay (ELISA R&D system). Plasma VEGF levels were measured before, 3 days and 4 weeks after TACE in 30 patients with HCC.  The development of metastasis was evaluated at the end of the third month after TACE.
RESULTS: The plasma VEGF levels of the 30 patients with HCC were 154.47±90.17 pg/ml. The total plasma VEGF levels after TACE increased compared with their basal levels (P<0.05), and the plasma VEGF levels had a tendency to increase in patients with heterogenous uptake of iodizdoil and portal vein thrombosis. Follow-up for six months showed metastatic foci in 20 patients (74%) with increased plasma VEGF, but none of the patients with decreased plasma VEGF developed metastasis.
CONCLUSION: Increased plasma  VEGF expression is associated with the development of metastasis in HCC after TACE.

BACKGROUND: Budd-Chiari syndrome  (BCS)  is a di\|sease caused by blood flow obstruction of the main hepatic veins (MHVs) and/or the outlet of the inferior vena cava (IVC), characterized by retrohepatic portal hypertension (PHT) and/or IVC hypertension. In the past decade, over 3000 cases of BCS have been reported in China. This study was to sum up our 20-year experience in surgical treatment of BCS and to investigate its pathological classification and principles of surgery.
METHODS: The data from 1360 BCS patients were analy\|zed retrospectively.
RESULTS: Four types (6 subtypes) were classified according to IVC angiography and hepatovenography: type Ⅰa (594 patients), type Ⅰb  (123),  type Ⅱ  (292),  type Ⅲa  (237),  type Ⅲb  (112),  and type Ⅳ  (2).  Surgical procedures included: improved splenopneumopexy  (265   cases),  finger or balloon membranotomy  (407),  radical resection of membrane and thrombus  (275),  IVC bypass (88: cavocaval transflow 71 cases, and cavoatrial transflow 17 cases), mesocaval C-shape shunt  (192),  splenocaval shunt  (32),  splenoatrial shunt  (23),  splenojugular shunt  (57),  mesoatrial shunt  (8),  and combined methods (6), including plenal-cavoatrial shunt  (4),  and mesocavoatrial shunt  (2),  splenorenal shunt  (4),  mesojugular shunt  (2),  and other  methods   (1).  The perioperative death rate and the complication rate after operation was  3.09% (42/1360) and  14.8% (201/1360) respectively. 885 cases were followed up from 9 months to 15 years  (average   6.8±1.2  years. The 791  (89.4%)  of 885 patients were successfully treated, 61 patients  (6.89%)  had a recurrence, and 33 died.
CONCLUSION: Surgical treatment of BCS is dependent on a correct diagnosis and classification of the disease.

BACKGROUND: Liver biopsy plays an important role in accurate diagnosis of various liver diseases in children and liver damages caused by systemic illnesses. This study was designed to evaluate the value of liver biopsy in diagnosis of liver diseases in children and explore the relationship between their pathological changes and clinical manifestations.
METHODS: One-second liver biopsy was performed in 1023 pediatric patients with liver diseases at our department from 1983 to 2000.  Diagnosis of viral hepatitis was based on the diagnostic criteria formulated by the Chinese Society of Infectious and Parasitic Diseases in 1995.  Inflammatory changes of the liver were graded from 0 to 4 (G0-4). 
RESULTS: Liver biopsy was performed successfully in 1020 patients including 135 infants and young children, of whom 90% were hospitalized patients with chronic liver diseases. Hepatitis virus was the leading cause for chronic liver diseases, among which hepatitis B was detected in  75.4% of the patients. Sixty-nine patients showed liver impairment induced by disorders relevant to that metabolism, Wilson’s disease, and glycogen storage disease. Liver inflammatory injury  (<G2) was found in 76.4% of the patients with hepatitis B in contrast to 61.2% of adult patients. It was aggravated with age in the patients with hepatitis B and C and peaked at their schooling age. Moderate or severe liver injuries were not seen in infants with chronic hepatitis B. Two infants had chronic hepatitis C. Patients with non-viral hepatitis showed specific, non-specific histological changes and liver cirrhosis.
CONCLUSIONS: Liver inflammatory injuries are more common in children with hepatitis B than in adult patients, and severe inflammatory changes are seen in children with hepatitis B and C at their school age. Liver injuries induced by non-viral factors seem to be increasing, and liver biopsy in children is safe and feasible in the diagnosis of liver diseases.

BACKGROUND: Portal hypertension is a common disease with a high mortality and serious effect on the life quality of patients. Presently, shunt and disconnection are commonly used for surgical treatment of portal hypertension. The aim of this study was conducted to analyze the results of a modified Sugiura procedure for the management of 160 cirrhotic patients with portal hypertension.
METHODS: The results of a modified Sugiura procedure for the treatment of 160 cirrhotic patients with portal hypertension from January 1991 to July 2002 were retrospectively analyzed.
RESULTS: The operative mortality for the procedure was zero. Postoperative intra-abdominal bleeding was noted in 2 patients, drowned lung in 1, pneumonia in 1, and splenic venous thrombosis in 4. Of the 160 patients, 157 (98%) were followed up from 6 months to 11.5 years. Of the 157 patients, only one died of hepatic coma 6 years after operation, and 3 of rebleeding. The absolute and relative survival rates were 97.5%(156/160) and 99%(159/160), respectively. The absolute and relative occurrence rates of hepatic coma were 2.5%（4/160）and 0.6%（1/157）, respectively. The absolute and relative occurrence rates of rebleeding were 3.8%（6/160）and 1.9%（3/157）, respectively. In 96 of 116 Child B patients (82.8%), liver function improved from preoperative class B to A 3 months after operation. Sixty-five patients were subjected to gastroscopy and 22 patients, esophageal barium photography 6 months after operation. Gastro-esophageal varices disappeared in 56 patients (64.4%, 56/87), obviously improved in 30 (34.5%, 30/87), and unchanged in 1 (1.2%, 1/87). The occurrence rate of portal hypertensive gastropathy (PHG) was 13.9%(9/65).
CONCLUSION: Our results showed that the modified Sugiura procedure is effective in the treatment of portal hypertension, with a low rate of operative complication, bleeding recurrence, and hepatic coma.

BACKGROUND: This study was to clarify the high risk factors for subphrenic infection (SI) after liver resection for patients with hepatic malignancy.
METHODS: Three hundred and sixty-eight patients who had undergone hepatectomy from January 1985 through June 2002 were randomly divided into 2 groups according to resection of liver parenchyma, hepatic cirrhosis, primary liver cancer, intraoperative blood loss, and subphrenic drainage. The chi-square was used for statistical analysis.
RESULTS: Thirteen patients (3.53%) of the 368 patients had SI. The high-risk factors for SI after hepatectomy were related to resection of liver parenchyma and hepatic cirrhosis; but the course or stage of primary liver cancer was not related to the incidence of SI. Intraoperative blood loss of over 1500 ml was found to be a significant risk factor for postoperative SI. Adequate drainage of the subdiaphragm and the raw surface of the liver after operation was essential to decreasing SI after liver resection.
CONCLUSION: Inadequate subphrenic drainage may largely contribute to SI in patients with hepatic malignancy undergoing hepatectomy apart from other factors. Comprehensive measures should be taken to prevent the infection after hepatectomy.

BACKGROUND: Malignant gastric carcinoids are often accompanied with liver metastasis synchronously or metachronously. Because of the slow growth rate of carcinoids, patients with metastatic tumors can undergo resection for potential cure or for symptom palliation. This study was designed to evaluate the clinicopathologic characteristics and the diagnosis and management of malignant gastric carcinoids. 
METHODS: Seven patients with malignant gastric carcinoids admitted to our hospital between 1990 and 2002 were followed up and reviewed retrospectively.
RESULTS: Liver metastases were found in all the patients, of whom 3 had lesions simultaneously and the other 4 had lesions postoperatively. More than 2 lesions were found in all these patients, except a solitary liver lesion in one. Follow-up showed two patients died within 2 months, three patients  in 20, 25 and 32 months after operation respectively, and the other two have been surviving for more than 5 and 3 years respectively.
CONCLUSIONS: Malignant gastric carcinoids have a high metastatic tendency to the liver. Surgical treatment in combination with other therapeutic approaches can significantly prolong the survival rate of the patients.

BACKGROUND: The molecular mechanism of hepatic metastasis of colorectal cancer is not well understood. The aim of this study was to assess the relations between phospholipid contents of cellular membrane and isoenzyme expression of protein kinase C (PKC) and their effects on hepatic metastasis of colorectal cancer.
METHODS: High performance liquid chromatography was used to detect contents of cell membrane phospholipids: phosphatidylinosital (PI), phosphatidylserine (PS), phosphatidylethanolamine (PE) and phosphatidylcholine (PC) in primary foci, paratumor mucosa and hepatic metastatic foci in patients with colorectal carcinoma. The mRNA expression levels of PKC-α, -βII, -δ, -ε, -λ, -ζ isoenzymes were detected with the QRT-PCR technique.
RESULTS: The levels of PI, PC and PE in primary foci and hepatic metastatic foci were higher than those in paratumor mucosa. The level of PE in hepatic metastatic foci was much higher than that in primary foci (t=98.88, P<0.01); but the levels of PI and PC were not significantly different between primary foci and hepatic metastatic foci (t=1.73, 1.36, P>0.05). The expression levels of PKC-βII, -δ, -ε, -λ, -ζ were enhanced in primary foci and hepatic metastatic foci, but the level of PKC-α in primary foci was decreased as compared with that in paratumor mucosa. The levels of PKC-δ, -ε, -λ, -ζ in hepatic metastatic foci were higher than those in primary foci. A positive correlation was observed between the expression levels of PI, PC and PKC-βII and also between those of PE and PKC-δ, -ε, -λ, -ζ. However, there was a close negative correlation between PE and PKC-α.
CONCLUSION: Increased levels of PI and PC and decreased ratio of PKC-α to PKC-βII are related to colorectal cancer genesis. Increased levels of PE, increased expression of PKC-δ, -ε, -λ, -ζ isoenzymes and decreased level of PKC-α are related to hepatic metastasis in colorectal carcinoma.

BACKGROUND:  Non-cirrhotic portal hypertension is a common cause of portal hypertension in developing countries. To understand its etiopathogenesis we developed an animal model by repeated portal endotoxemia induced through the gastrosplenic vein.
METHODS: Twenty-nine rabbits (1.5-2.0 kg) were divided into control (group I, n=13) and experimental (group II, n=16) groups. Heat killed E.coli were injected through an indwelling cannula into the gastrosplenic vein in pre-sensitized animals. The animals were sacrificed at 1, 3 and 6 months.
RESULTS: The mean portal pressure in group II animals was significantly (P<0.05) higher than in  group I at 1 (17.5±3.4 vs 10.4±2.2 mmHg), 3 (17.8±1.3 vs 7.2+3.6 mm Hg), and 6 (19.8±3.1 vs 10.3±4.8 mmHg) months. Similarly, the mean splenic weight in group II was significantly greater than in group I (P<0.05). Histopathologically, the  spleen showed medullary congestion, hemosidrin-laden macrophages and mild fibrosis. Histologically, the liver had normal parenchyma with mild portal lymphocytic infiltrates and kupffer cell hyperplasia. No significant anomalies were detected by liver function tests.
CONCLUSIONS: The rabbit model showed significant splenomegaly with a persistent  increase in portal pressure and mild fibrosis without hepatic parenchymal injury, quite akin to non-cirrhotic portal fibrosis as seen in humans. Recurrent intra-abdominal infection may play an important role in the pathogenesis of non-cirrhotic portal fibrosis.

BACKGROUND: Viral hepatitis is considered a major public health problem in most areas of the world. In acute and chronic infections, hepatitis D virus (HDV) infection often leads to a more severe disease.This study was designed to prepare microarrays for HDV detection.
METHODS: The specific primers of PCR were designed according to the conserved region of HDV. The cDNA microarrays were prepared by spotting PCR products onto the surface of glass slides by robotics. Restriction display PCR (RD-PCR) was used to label the samples.
RESULTS: Sequences were aligned, and the results showed that the products of PCR amplification were the specific gene fragments of HDV. Hybridizing signals on gene chip showed the specificity and sensitivity in detecting HDV were satisfactory.
CONCLUSION: Using PCR amplified products to construct gene chips for clinical diagnosis of  HDV is a quick, simple and effective method.

BACKGROUND: In orthotopic liver transplantation, ischemic-reperfusion is one of the most important factors that cause the incidence of biliary complicance. The aim of this study was to investigate the effects of ischemia reperfusion on epithelial cells apoptosis and proliferation of intrahepatic bile duct (IBD) (>20 μm).
METHODS: 30-minute warm ischemia was applied to rat livers respectively, and experiment was performed on days 2, 7, 14, 28 after reperfusion. Apoptosis was determined in situ by morphology and TUNEL, and cholangiocyte proliferation was evaluated in situ by morphometry of liver sections stained for cytokeratin-19  (CK-19) and by proliferating cellular nuclear antigen staining in liver sections.
RESULTS: Two days after ischemia reperfusion, apoptosis of cells was observed in large intrahepatic bile ducts (>20 μm) (5.6%±1.2%), but the number of large intrahepatic bile ducts reduced (0.32±0.06). Seven days after ischemia reperfusion, the apoptosis index of cholangiocytes decreased to 1.2%±0.3%, and the number of intrahepatic bile ducts began to proliferate and returned to nearly normal on day 28.
CONCLUSION:  Ischemia reperfusion  causes a decrease in the number of intrahepatic bile ducts (>20 μm) as a result of a higher rate of apoptosis and absence of initial proliferation.

BACKGROUND: This study was designed to assess the roles of oval cells and c-myc mRNA in the process of hepatocarcinogenesis and to clarify the function of carcinogene c-myc in the development of hepatocellular carcinoma (HCC) and the mechanism of inhibitory function of uscharidin on HCC in mouse hepatocarcinogenesis.
METHODS: A total of 120 clean SD mice were divided into normal group, cancer induction group, and intervention group. The normal group was fed with standard forage while the rest two groups were given p-dimethylaminoazobenzene (DAB) to induce cancer. Thirteen weeks after induction of cancer, the two groups were fed with standard forage and water. Once the pattern was set up, the intervention group was given uscharidin injection into the abdominal cavity from the first week to the 14th week. On the 2nd, 4th, 6th, 8th, 10th, 12th, 14th, 16th, 18th, 20th, 22nd, and 24th week, all mice were killed and biopsied from the liver lobe for pathological analysis. At the same time, the number of tumor nodes was counted and the expression of c-myc mRNA was tested by RT-PCR.
RESULTS: Since the 2nd week after cancer induction, proliferated oval cells could be seen in the portal area. Initially, the oval cells appeared in the cortical layer of the portal area, then proliferated gradually and immigrated into the liver parenchyma. In the period of fibrosis after liver proliferation, proliferated heaps of oval cells were noted in both portal and peripheral areas. In the period of carcinomatous change, oval cells could be seen both outside and inside of cancer nodes, but most of them were distributed outside. The c-myc gene was expressed negatively in the liver tissue of mice. The quantity of the expression began to increase at the time of infection of the liver and tended to increase with the degree of hepatic injury. In the period of canceration, the expression level of c-myc mRNA increased gradually. The intervention of uscharidin could not inhibit but delay the increase of the expression of c-myc mRNA.
CONCLUSION: Oval cells are closely related to hepatocarcinoma cells, which play an important role in the occurrence and development of hepatocarcinogenesis. Uscharidin can inhibit the occurrence of hepatocarcinogenesis or local spreading at the early stage of cancer induction by DAB, but it cannot inhibit the expression of c-myc.

BACKGROUND: The inoculation of plasmid DNA encoding tumor-associated antigens is a novel and powerful strategy for antitumor vaccination. This study was designed to construct the DNA vaccine of mouse AFP and to observe the specific cellular immunologic responses and the antitumor responses in mice induced by this vaccine.
METHODS: The murine AFP gene was amplified by RT-PCR from total RNA extracted from Hepa1-6 cells and cloned into the vector pcDNA3.1 to construct pmAFP.The DNA vaccine was identified by restriction enzymeanalysis, sequencing and expression. EL-4 (mAFP) was developed by stable transfection of EL-4 cells with pmAFP. The frequency of cells producing IFN-γ in splenocytes harvested from the mice immunized with the DNA vaccine by intramuscular injection was measured by enzyme linked immunospot (ELISPOT).  The mice immunized with the DNA vaccine were inoculated with EL-4 (mAFP) cells in back to observe the inhibitory effect of the immunization on tumor. On the other hand, blood samples were collected from the immunized mice to check the functions of the liver and kidney.
RESULTS: The murine AFP gene was successfully cloned by RT-PCR. Results from restriction enzyme analysis, sequencing and expression showed that the DNA vaccine was successfully constructed. The expression of mAFP mRNA in EL-4 (mAFP) was confirmed by RT-PCR.  The results of ELISPOT showed that the number of IFN-γ- producing  cells of the pmAFP  vaccine group was significantly higher than that of other groups (P<0.01). The tumor volume in the pmAFP vaccine  group （1042.42±123.71 mm3） was significantly smaller than that in other groups (P<0.01). The function of mouse liver and kidney in each group was unchanged.
CONCLUSION: The successfully constructed DNA vaccine of AFP can induce specific cellular immunologic responses and significant antitumor reponses in mouse and has no impact on the function of mouse liver and kindey.

BACKGROUND: It is the key point for vascular endothelial growth factor (VEGF121) gene related therapy as to how to transfect and express the gene safely, effectively and repeatedly. This study was designed to investigate the VEGF121 transfection and expression in primary cultured rat hepatocyte.
METHODS: After construction of vector internal ribosome entry site-enhanced yellow fluorescent protein (pIRES-EYFP)/VEGF121, the transfection and expression of the exogenous VEGF121 gene in primary cultured rat hepatocytes were observed through RT-PCR, Western blot and fluorescent microscopy.
RESULTS: pIRES-EYFP/VEGF121 plasmid was constructed and transfected successfully into primary cultured rat hepatocytes, the transfection and expression of  gene in primary cultured rat hepatocytes were examined by RT-PCR and Western blot, and yellow-green fluorescence was observed through a fluorescent microscope.
CONCLUSION: The successful transfection and expression of plasmid pIRES-EYFP/VEGF121 in primary cultured rat hepatocytes provides a foundation for hepatocyte transplantation and gene therapy after modification of hepatocytes by the gene.

BACKGROUND: Some reports have confirmed that occurrence and development of tumor are related with lots of oncogene, anti-oncogene and cytokine. This study was to detect the expression of TNFmRNA, tumor necrosis factor (TNF) and TNF receptor (TNFR) in the gallbladder mucosa which developed from hyperplasia, dysplasia to carcinoma, and to further discuss the pathogenecity of gallbladder carcinoma.
METHODS: The expression of TNFmRNA, TNF and TNFR was detected by in situ hybridization and immunohistochemistry on the surgical specimens of hyperplasia, dysplasia and carcinoma of the gallbladder.
RESULTS: The percentage of positive cells expressed TNFmRNA in the gallbladder mucosa was 0%, 20%, and 90%, respectively in hyperplasia, dysplasia and carcinoma (P<0.05). The percentage of positive mononuclear cells (MNCs) expressed TNFmRNA in hyperplasia, dysplasia and carcinoma of gallbladder tissues was 15%, 85%, and 90%, respectively (P<0.05). The number of positive MNC high-power field (Hpf) was 4.85±1.50, 6.00±2.71, and 9.33±3.07, respectively (P<0.05). The number of carcinoma cells and MNCs expressed by TNFmRNA per high-power field was increased with the increase of tumor stage. The number of carcinoma cells/Hpf in stages I-III and IV-V  was 9.13±4.39 and 7.13±2.53 (P<0.05), and that of MNC/Hpf was 14.80±4.02 and 11.10±2.23 (P<0.01). The number of carcinoma cells and MNCs expressed by TNFmRNA per Hpf was increased with the increase of tumor size. In tumors of more than 2 cm or less than 2 cm in diameter, the number of positive carcinoma cells/Hpf was 14.00±4.20 and 8.83±4.96, respectively (P<0.05), but that of MNC/Hpf was 10.50±2.54 and 7.00±2.83 (P<0.05). The pattern of TNF protein expression was similar to that of TNFmRNA, whereas TNF protein expression was more frequent and extensive than TNFmRNA expression. TNFR was expressed in endothelial cells and MNCs of carcinoma, and was negative in mucosal epithelial cells and tumor cells. A positive linear correlation in TNFmRNA expression was observed between tumor cells and MNCs (r=0.687, P<0.01), a correlation in TNFmRNA and TNF protein expression of tumor cells (r=0.847, P<0.001), and a correlation in TNFmRNA and TNF protein expression of MNC in tumor tissue (r=0.643, P<0.01).
CONCLUSIONS: TNFmRNA and TNF protein expression is increased during the development of gallbladder mucosa from hyperplasia, dysplasia to carcinoma, and is increased with tumor stage. This finding suggests that TNF is involved in the pathogenesis of gallbladder carcinoma induced by gallstones and the TNF expression in cancer cells may serve as a marker for tumor stage.

BACKGROUND: Hilar cholangiocarcinoma has a low radical resection rate and a poor long-term survival rate. In recent years, its prognosis has been improved with advancement  of preoperative diagnostic techniques and surgical techniques. The aim of this study was to  evaluate the prognostic factors of hilar cholangiocarcinoma and the relations of surgical procedure to the prognosis of the carcinoma.
METHODS: A retrospective cohort study was done in 198 patients with hilar cholangiocarcinoma (117 men and 81 women, aged from 27 to 81 years), who had been  admitted to this hospital from December 1997 to December 2002. Their symptoms were jaundice (94.5％), pruritus （56.6％） and abdominal pain （33.8％）. Bismuth-Corlette classification showed type I in 14 patients, type II in 19, type IIIa in 12, type IIIb in 15, type IV in 112, and unclassifiable type in 26. 144 patients underwent laparotomy and  others received bile drainage endoscopically (including endoscopic retrograde biliary drainage (ERBD) or endoscopic metal biliary endoprosthesis (EMBE) in 21 patients, endoscopic nose-biliary drainage (ENBD) in 31 or percutaneous transhepatic cholangiodrainage in 2. 120 patients (83.3%) received tumor resection including radical resection in 59 patients (41.0%). Twenty-three patients  underwent paunched biliary exploration and drainage.
RESULTS: Cox’s regression model analysis showed that occupation, preoperative total serum bilirubin level, operative procedure and postoperative adjuvant radiation were significantly related to postoperative survival rate in contrast to gender, age, choledocholithiasis, hepatitis, preoperative serum CA19-9 level, Bismuth-Corlette type, histopathologic grading and postoperative chemotherapy. The survival of patients in groups of biliary drainage, palliative resection and radical resection differed statistically and prolonged in a descending order. No statistical difference was found between ERBD or EMBE group and palliative resection group. So was between ERBD or EMBE group and biliary drainage group, or between ENBD group and biliary drainage group. The survival differed statistically between ERBD or EMBE group and ENBD group.   
CONCLUSIONS: Operative procedure is the most important prognostic factor affecting the operative results of hilar cholangiocarcinoma. Radical resection is still the primary measure for a cure and long-term survival of the patients. For patients with irresectable hilar cholangiocarcinoma, no evidence has shown that the prognosis after treatment of  ERBD or EMBE is poorer than that after laparotomy.

BACKGROUND: Severe acute pancreatitis (SAP) as a common acute abdomen due to complicated causes is characterized by lots of morbidities, difficult treatment, and high mortality. This study was designed to investigate the role of individually staged nutritional support (ISNS) in the treatment of SAP.
METHODS: One hundred patients with SAP admitted to our hospital from January 1997 to October 2002 were randomly divided into total parenteral nutrition group (TPN group, 50 patients) and individually staged nutrition group (individualized group, 50 patients), between which the therapeutic outcome and the incidence of complications were carefully analyzed.
RESULTS: Compared with the TPN group, the individualized group had less complications (62 vs 94 patients) including incubation related complications (2% vs 16%), superinfections (8% vs 30%), hepatic functional insufficiency (4% vs 24%) and intra-peritoneal infections (4% vs 12%), in addition to a sooner restoring of oral nutrition (18.5 vs 24.8 days, P<0.05), a shorter hospital stay (24.5 vs 30.2 days) and a lower hospital cost (4.1 vs 5.8 10 000 yuan, P<0.05).
CONCLUSION: ISNS, which provides SAP patients with sufficient energy and nutrients according to their true pathological status, is an ideal nutrition planning with lowered incidence of complications, shortened hospital stay and lightened financial burden.

BACKGROUND: Although a variety of tumor markers are available for diagnosis of pancreatic cancer, their sensitivity and specificity have not yet been ideal. The aims of this study was to detect a panel of serum tumor markers and to evaluate their significance in the diagnosis and prognosis of pancreatic cancer patients.
METHODS: Eight serum tumor markers including AFP, CEA, CA-50, CA72-4, CA-125, CA153, CA19-9 and CA242 were detected in 129 patients with pancreatic cancer by usingchemiluminescence immunoassay, immunofluorescence assay and immunoradiometric assay, respectively. The levels of these markers were compared in 99 patients with non-pancreatic malignant tumor, 63 patients with other benign diseases, and 27 patients with pancreatic cancer after pancreatectomy.
RESULTS: Among the 8 tumor markers, CA19-9, CA242, CA-50, and CA72-4 were more sensitive in the diagnosis of pancreatic cancer. Parallel combined testing could increase the diagnostic sensitivity to 89.2%, and serial combined examination could increase the diagnostic specificity to 92.3%. The serum tumor markers levels were decreased significantly after radical tumor resection.
CONCLUSIONS: Serum CA19-9, CA242, CA-50, and CA72-4  are the preferred tumor markers to be used in the diagnosis and follow-up of operated cases of pancreatic cancer. Testing of a panel of multiple serum tumor markers may increase the sensitivity and specificity in the diagnosis of pancreatic cancer.

BACKGROUND: Nonfunctioning islet cell tumor (NIT) as a rare pancreatic endocrine neoplasm is characterized by unspecific clinical symptoms and is hard to diagnose. In China, NIT accounts for 15%-41% in pancreatic endocrine neoplasms just next to insulinoma. In this study, we evaluated the surgical modalities of NIT.
METHODS: From January 1978 through February 2002, 41 patients with NIT were treated at the Department of Surgery of the First Affiliated Hospital, China Medical University, Shenyang, China. Tumors in the head of the pancreas were noted in 28 patients, and in the body or in the tail in 13 patients. The mean diameter of the tumors was 10.7 cm. Fifteen patients underwent enucleation and 21 received pancreatectomy. Tumors were unresectable in 5 patients because of extensive infiltration. The mean diameter was 9.6 cm in patients treated by enucleation, 13.1 cm in those by pancreaticoduodenectomy, 9.9 cm in those by distal pancreatectomy, and 11.6 cm in those with unresectable tumors.
RESULTS: The curative resection rate was 88% (n=36), and the complication rate after enucleation and pancreatectomy was 33% (n=5) and 14% (n=3), respectively. No local recurrence was found after both enucleation and pancreatectomy. Liver metastases occurred in 3 patients treated by enucleation.
CONCLUSIONS: Both enucleation and pancreatectomy are effective for NIT of the pancreas. No local recurrence has been found in patients treated by the two surgical procedures. The complication rates of the two modalities are comparable.

BACKGROUND: Pancreatic endocrine tumors are uncommon neoplasms and can lead to systemic disorder including glucagonoma syndrome, a very rare prototypical paraneoplastic phenomenon. The aim of this study was to assess the diagnosis and surgical strategy for the treatment of glucagonoma syndrome.
METHODS: The clinical data of a case of pancreatic head tumor with typical glucagonoma syndrome of necrolytic migratory erythema (NME), diabetes mellitus (DM), anemia, and glossitis were retrospectively analyzed.
RESULTS: Cutaneous eruption occurred mainly in the groin, extremities, thighs, buttocks, and perineum. A highly elevated level of serum glucagon was detected by radioimmunoassay. A tumor located in the head of the pancreas was well-defined by pre and intra-operative ultrasonography, contrast enhanced computed tomography, and magnetic resonance imaging. Tumor enucleation was performed, showing significantly improved symptoms. Near complete resolution of NME was shown one week after surgery.  Surgical complications or recurrence was not found.
CONCLUSIONS: The diagnosis of glucagonoma syndrome is established by marked clinical features such as NME as the hallmark clinical finding, hyperglucagonemia, and radiographically demonstrated neuroendocrine tumor. The topographic diagnosis of glucagonoma can be achieved by combined imaging methods. Enucleation of tumor is a valuable treatment for solitary pancreatic tumor without peripancreatic invasion, liver metastasis, and pancreatic duct compression.

ABSTRACT: Fulminant hepatic failure is a medical emergency. When this condition declared itself irreversible, a timely liver transplantation is the only effective treatment. A 34-year-old Chinese with fulminant hepatic failure was evaluated as a potential liver transplantation candidate. On the erect chest radiograph, Chilaiditi’s sign has developed over a very short period of a week due to rapid shrinkage of the liver. Awareness of Chilaiditi’s sign facilitated distinguishing the condition of free gas under the diaphragm due to bowel perforation and subphrenic abscess by gas forming micro-organisms. Rapidity of onset of this sign parallels the deterioration of liver function and reflects the urgency of condition.