OBJECTIVE: To review the current progress of islet cell transplantation in patients with insulin-dependent diabetes, emphasizing on the difficulties with recovering and preserving islet cell mass and function, 30% of which is lost during the peri-transplantation period.
RESULTS: The islet-cell isolation technique is perfected, but improvements are still progressing in two major directions: preservation of islet cells and tolerance induction. Optimum islet cell viability and function depends on appropriate revascularization of the islet graft and blockade of thrombus formation as well as cytokine and free radical release. Conditioning the islet cells in-vitro prior to transplantation to either upregulate VEGF expression or downregulate NF-kappa B transcription factor has proven to improve revascularization and to prevent islet cell apoptosis and cytokine-mediated damage. Tolerance induction is currently being best achieved by selecting and combining immunosuppressive agents such as monoclonal antibodies which target the major signaling molecules during immune activation, but which are least toxic to islet cells.
CONCLUSIONS: Patients with insulin-dependent diabetes will greatly benefit from current developments in effective approaches to protect islets during the peritransplant period. Emerging interest in stem cell biology and differentiation may provide the ultimate solution to the problem of organ scarcity and islet cell protection from the peritransplant induced damage.

ABSTRACT: Nuclear factor-kappa B (NF-κB) as an essential transcription factor in the control of expression of the cytokine-induced genes in immune and inflammatory responses regulates cytokines in allograft rejection. In this review, we summarize the general properties of NF-κB and the principal findings to shed new light on transplantation.

OBJECTIVE: To investigate the alloimmunogenicity of liver specific antigen and its effects on allolymphocytes.
METHODS: Liver specific antigen isolated from inbred F344 rats was used as immunogen to immunize inbred Lew rats through different immunization pathways such as low-dose long-term hind footpad, high-dose portal vein and thymus immunization. Western blotting, DNA fragments gel electrophoresis, mixed lymphocyte culture (MLC) and mixed lymphocyte hepatocyte culture (MLHC) were employed to analyze the immune state after immunization.
RESULTS: At the time point of sampling, different degree of specific low immunoresponses appeared in all immunized groups as well as cyclophosphamide (CY) treated group. Compared with group Ⅰ, other groups expressed caspase-3 significantly as detected by using Western blotting. DNA fragment gel electrophoresis of splenocytes showed lymphocyte apoptosis. Compared with the group Ⅰ, MLC of the experimental groups showed no significant changes except that of the group Ⅴ, whereas MLHC decreased markedly (P<0.05).
CONCLUSIONS: Liver specific antigen not only has alloimmunogenicity to induce alloimmunoreaction but induce antigen specific low immunoresponses and antigen specific lymphocyte apoptosis by high-dose or low-dose long-term immunization. It may be an important transplantation antigen that may lead to a novel way to liver transplantation immunotolerance.

OBJECTIVE: To explore the changes of mitogen activated protein kinase (MAPK) cascade pathway and anti-stress response of hepatocytes after liver transplantation.
METHODS: Ten normal liver specimens and 18 punctured donor liver specimens were divided into 3 groups: A (control∶10), B (no rejection∶10) and C (acute rejection∶8). MAPK, Ras, Jun and heat shock protein 70 (HSP70) were tested immunohistochemically while Ras and HSP70 were tested by in situ hybridization. All sections were subjected to image analysis.
RESULTS: Protein expressions of MAPK, Ras and Jun were increased by an ascending order of groups A, B and C. The protein expression of HSP70 was the highest in group B but lower in group C. Expressions of Ras and HSP70 mRNA were consistent with those of protein.
CONCLUSIONS: The changes of the MAPK cascade pathway and anti-stress response of hepatocytes after liver transplantation are one of regulation mechanisms for protecting the hepatocytes from damage after liver transplantation. This mechanism is active to support individual survival.

OBJECTIVE: To investigate the dynamic alternations of HBV markers of active HBV replication recipients receiving lamivudine prophylaxis after liver transplantation.
METHODS: Serial liver biopsy samples and sera were obtained from 15 recipients and examined with enzyme-linked radioimmunoassay for HBsAg, HBeAg, HBsAb, HBcAb and HBeAb, and fluorescent quantitative assay for quantitation of HBV DNA in serum. Immunohistochemical staining of HBsAg, HBcAg and HBV DNA hybridization in situ were used to detect HBV markers in liver biopsy samples.
RESULTS: 100 mg lamivudine taken orally every day for 2 weeks before transplantation enabled 12 (80%) of 15 active viral replication recipients (HBV DNA positive) to converse to HBV DNA negative. HBsAb, HBcAb and HBeAb in serum emerged in 1-2 weeks after liver transplantation, and disappeared gradually within 6 months; HBV DNA fluorescent quantitative assay showed constant negativity in serum. Immunohistochemical staining of HBsAg, HBcAg and HBV DNA hybridization in situ in liver biopsy samples showed negative results synchorously. Eight of the 15 HBV active replication recipients lost HBV markers thoroughly both in serology and tissue staining as well as HBV DNA hybridization in situ of serial liver biopsy samples from 12 to 44 weeks after liver transplantation. Should any of HBsAg, HBeAg in serology and HBsAg, HBcAg in immunohistochemical staining was positive, or HBV DNA detectable in serum, or HBV DNA hybridization in situ in liver tissue positive, allograft HBV reinfection or De novo liver allograft infection could be diagnosed. Furthermore, if associated with elevation of ALT and bilirubin, the diagnosis of HBV hepatitis recurrence could be established.
CONCLUSION: Allograft HBV reinfection or De novo liver allograft infection in active viral replication recipients could be prevented with lamivudine regimen, and further clearance of HBV may be possible if proper measures are taken.

OBJECTIVE: To introduce a new surgical technique—cavoportal hemitransposition in liver transplantation.
METHODS: Since Tzakis first reported liver transplantation with cavoportal hemitransposition in the presence of diffuse portal vein thrombosis in 1998, five relevant research papers have been published, in which a total of 23 cases were treated.
RESULTS: Of the 23 cases, 11 underwent conventionally cavoportal anastomosis as an end-to-end anastomosis, 8 renoportal anastomosis, and 4 end-to-side anastomosis preceded by piggyback. Eight patients died after operation. Fifteen patients are still alive with normal liver and kinase function, with a longest follow-up of 37 months. They enjoy a normal life without dietary or other restriction.
CONCLUSION: Portal vein thrombosis is no longer an absolute contraindication to liver transplantation. This new technique is useful and reliable for liver transplantation.

OBJECTIVE: To summarize the experience in modified reconstruction of the hepatic outflow tract during piggyback liver transplantation at our hospital.
METHODS: The clinical data on 67 patients undergoing piggyback liver transplantation with modified hepatic outflow tract reconstruction from January 1999 to October 2002 were analyzed retrospectively.
RESULTS: In this group, 7 patients (10.45%) died perioperatively. Complications included: pulmonary infection (38 patients); multiple organ system failure (10), intraperitoneal bleeding (6), acute respiratory distress syndrome (14), thrombosis of the hepatic artery (1), and bile leakage (1). No hepatic outflow occluded. Two recipients survived for over 3 years, 8 over 2 years, and 19 over a year.
CONCLUSION: Modified hepatic outflow reconstruction in piggyback liver transplantation may increase the success rate of liver transplantation and decrease technical complications.

OBJECTIVE: To investigate the influences of helicobacter pylori (Hp) infection, liver function, gastroesophageal varicosity, esophageal varicosity ligation (EVL), and esophageal varicosity sclerotherapy (EVS) on patients with portal hypertensive gastropathy (PHG).
METHODS: Fourty-seven patients with liver cirrhosis included 34 patients with PHG and 13 patients without PHG. Liver function, Hp infection, and gastroesophageal varicosity were investigated clinically in all patients, and gastroscopy was made again in patients with EVL 1-2 months after operation to observe the changes of PHG.
RESULTS: The rate of Hp infection was lower in patients with liver cirrhosis than in controls. There was no relationship between Hp infection and PHG. The patients with gastroesophageal varicosity had a high incidence of PHG.
CONCLUSIONS: Despite no relationship between Hp infection and PHG, liver dysfunction can affect and promote PHG. Gastroesophageal varicosity may be closely related to PHG, but their degrees are not related. PHG can be caused or promoted by EVL.

OBJECTIVE: To evaluate the effect of tamoxifen (TAM) combined with a somatostatin analogue, octretide (OCT) on advanced liver cancer and whether tamoxifen combined with OCT is superior to regular chemotherapeutic agents 5-Fu and mitomycin C (MMC).
METHODS: Thirty-nine patients with inoperable liver cancer were randomly subdivided into TAM+OCT group (n=24) and regular chemotherapeutic group (n=15). They received treatment for three months respectively. Blood cell count, liver function, immunologic function, blood α-FP was regularly measured. Liver lump and extrahepatic metastasis were examined by CT. The patients were followed up after treatment and conducted survival analysis.
RESULTS: In the TAM+OCT group, complete response is 4 patients, partial response is 7 patients, no change is 9 patients and progressive disease is 4 patients; blood level of ALT and AST had no noticeable change, IgE and IgG increased (P<0.01), and α-FP lowered (P<0.05). In regular chemotherapeutic group, no change is 4 patients and progressive disease is 11 patients. There was conspicuous statistical difference in the two groups. The accumulative survival rates of 6 months, 1 year and 2 years were 95.7% vs 41.2% (P<0.01), 63.7% vs 21.1% (P<0.01), 25.4% vs 0 (P<0.01), respectively. Medium survival time was 12.8 months in TAM+OCT group and 5.5 months in chemotherapeutic group.
CONCLUSIONS: TAM+OCT excerts reliable therapeutic effect on patients with inoperable ER(+) hepatocellular cancer. It is superior to 5-Fu and MMC in increasing the survival rate, prolonging survival time, and reducing side-effects.

OBJECTIVE: To study the effect of preoperative selective portal vein embolization (SPVE) in the two-step hepatectomy for patients with primary hepatocellular carcinoma (HCC) in injured livers.
METHODS: Twenty-six patients with HCC and cirrhosis who were not suitable for curative hepatectomy were treated by ultrasound-guided percutaneous transhepatic SPVE with a fine needle. The success rate, side-effects and complications of SPVE, serial changes of hepatic lobe volume and rate of two-step curative hepatectomy after SPVE were observed.
RESULTS: SPVE was performed in 24 patients (92.3%). In patients whose right portal vein branches were embolized, the right hepatic volume decreased but the left hepatic volume increased gradually. The ratio of the right hepatic volume to the total hepatic volume decreased from 64.0% before SPVE to 60.8% after l week, 55.1% after 2 weeks and 52.7% after 3 weeks, respectively. The side-effects included different degree of pain in the liver quandrant (17 patients), lower fever (9), and nausea and vomiting (7). The levels of aspartate alanine transaminase (AST), alanine transaminase (ALT) and total bilirubin (TBIL) increased after SPVE, but returned to the preoperative levels in 1week. After 2-4 weeks, two-step curative hepatectomy for HCC was performed in 13 patients (54.2%).
CONCLUSIONS: Ultrasound-guided percutaneous transhepatic SPVE with a fine needle is feasible and safe. It can extend the indications of curative hepatectomy for HCC in injured livers, and increase the safety of two-step hepatectomy.

OBJECTIVE: To characterize the clinical and radiographic findings in patients with pylephlebitis and liver abscess with an emphasis on the findings that help to differentiate this disorder from portal vein occlusion associated with hepatocellular carcinoma.
METHODS: We analyzed the clinical findings and radiographic images of four patients with pylephlebitis and liver abscess(es) who had been misdiagnosed as having hepatocellular carcinoma with portal vein thrombosis. Their medical records were reviewed in terms of clinical presentation, physical findings, laboratory data, treatment, and follow up.
RESULTS: All patients undergoing color duplex ultrasonography had an echogenic thrombus within an expanded portal vein with negative color-flow findings within the thrombus. Contrast enhanced CT in all the patients demonstrated portal vein thrombosis associated with “liver masses”. An intra-abdominal site of infection responsible for the subsequent ascending infection of the portal vein and liver was not identified in any patient on initial CT scan. At presentation, all patients were febrile and three of them had an elevated white blood cell count as well. All patients showed abnormalities of liver function.
CONCLUSIONS: Liver abscess(es) associated with pylephlebitis may mimic hepatocellular carcinoma with portal vein thrombosis. Clinical features that help to distinguish the two entities include presence or absence of fever, elevated white blood cell count, elevated alpha-fetoprotein, cirrhosis, and risk factors for hepatocellular carcinoma.

OBJECTIVE: To investigate the correlation between HLA class II molecules and the different outcome of viral hepatitis B.
METHODS: Thirty patients with chronic hepatitis B and 56 subjects who had spontaneously recovered from HBV infection in Zhejiang were enrolled in this investigation. HLA class II molecules types and alleles were determined by PCR-ssp.
RESULTS: HLA-DR12 was found in 21 of the 56 subjects (38%) recovered from hepatitis B, compared to 3 of the 30 patients with chronic hepatitis B [10%, relative risk (rr), 0.19; Pcorr<0.025] . The frequency of the allele of HLA-DR12 DRB1*1201 was higher in the subjects recovered from hepatitis B infection (32%) than in the patients with chronic hepatitis B (3%; rr, 0.07; Pcorr<0.005). On the contrary, more HLA-DR9 was detected in the patients with chronic hepatitis B (43%) than in the subjects who recovered from hepatitis B infection (18%; rr, 3.52; Pcorr<0.025). HLA-DQ9 was also detected with a higher frequency in the patients with chronic hepatitis B (43%) than in the subjects who recovered from hepatitis B (20%; rr, 3.13; Pcorr<0.05).
CONCLUSIONS: The HLA class II molecule DR12 and its allele DRB1*1201 are associated with protection against chronic hepatitis B in Zhejiang Province, China. HLA-DR9 and DQ9 are associated with chronicity of HBV infection.

OBJECTIVE: To study the preparation of diagnostic gene chip for detecting hepatitis B virus (HBV) and hepatitis C virus (HCV) and its accuracy in detecting HBV DNA and HCV RNA in serum and liver tissues.
METHODS: The probes, which depend on the conservative gene fragment of hepatitis virus, was designed, synthesized and spotted on the modified glass. The probes and some other control probes were assembled on the diagnostic microarray of hepatitis virus. The gene of hepatitis virus, purified from blood or tissue, was labeled with fluorescence and hybridized to the microarray. The hybridized microarry was scanned with microarray scanner and the diagnostic result was analyzed from the scanning data. Fourty patients with hepatitis B virus and 40 healthy people or 40 patients with hepatitis C virus were subjected to detection of HBV DNA and HCV RNA with the hepatitis virus gene chip by the double-blind method. Paraffin liver specimens obtained from 99 cases of posthepatitic cirrhosis were used to detect HBV DNA. The liver tissues and serum from 15 cases of chronic hepatitis B were used to detect HBV DNA. Simultaneously, HBsAg and HBcAg were detected in the serum by fluorescence microparticle quantitation, HBV DNA and HCV RNA in the serum by PCR, and HBcAg in liver tissues by immunocytochemistry or HBV DNA by in situ molecular hybridization.
RESULTS: Chip detection of serum specimens showed that 30 patients were HBV DNA positive and 10 HBV DNA negative in the 40 patients with HBV positive, 25 patients were HCV RNA positive and 15 patients were HCV RNA negative in the 40 patients with HCV positive, and all were HBV and HCV negative in the 40 healthy people. In 15 patients with HBV marker positive who were subjected to liver biopsy, 15 patients were detected HBV DNA positive in serum by gene chip, 15 patients HBcAg positive in liver tissues by immunocytochemistry, 14 patients HBV DNA positive in liver tissues by in situ molecular hybridization, and 14 patients HBV DNA positive in liver tissues by gene chip. Paraffin liver tissues specimens from the 99 patients with posthepatitis B cirrhosis showed that 67 patients were detected HBcAg positive by immunocytochemistry, 53 patients HBV DNA positive by in situ molecular hybridization, and 46 patients HBV DNA positive by gene chip. In the 46 patients, 40 patients were detected HBV DNA and HBcAg positive by in situ molecular hybridization and immunocytochemistry, 6 patients only HBcAg positive, and 33 patients HBcAg negative.
CONCLUSIONS: The designed diagnostic gene chip can be used to simultaneously detect serum HBV DNA and HCV RNA, but the positive rate of HCV RNA diagnosed by this chip is lower. The gene chip can detect HBV DNA in serum and in liver tissue.

OBJECTIVE: To establish a rat model of liver cancer complicated by liver cirrhosis and explore the effects of the spleen on immune function in this model.
METHODS: Liver cirrhosis was inflicted in rats by percutaneous injection of 40% CCl4 on the back. Walker-256 tumor cells were inoculated in the cirrhotic liver and splenectomy was performed. Two weeks later, the growth and metastasis of tumor were observed and the amount of ascites and the activity of NK cells and CD25 cells were investigated.
RESULTS: The amount of ascites and tumor volume were significantly higher in splenectomy group than in controls (P<0.01). Two weeks after inoculation, the activity of NK cells in both groups was decreased as compared with that before the inoculation (P<0.01). There was no significant difference between the two groups before and after the inoculation (P>0.05). The number of CD25 in both groups was higher than that before the inoculation (P<0.01). However, there was no significant difference between the two groups before and after the inoculation (P>0.05).
CONCLUSIONS: Splenectomy in early stage of tumor inoculation can stimulate the tumor growth and metastasis. The activity of NK cells and the number of CD25 are inhibited by tumor itself, not by splenectomy.

OBJECTIVE: To explore the feasibility of cloning of the hepatocyte receptor interacting with the Pre S1 protein of HBV by two-hybrid system.
METHODS: Yeast expression plasmids encoding fusion proteins of full length or portions of Pre S1 of HBV and DNA binding domain of yeast protein GAL4 were constructed and used to transform yeast reporter strain SFY526. Reporter gene product β-galactosidase activity was assayed as a measure of transcriptional activation in yeast. Mammalian expression plasmid encoding fusion proteins of full length Pre S1 and DNA binding domain of GAL4 was constructed and used to cotransfect hepatoma cell line Huh-7 together with CAT reporter plasmid. Cell extracts were assayed for CAT activity by thin-layer chromatography.
RESULTS: The fusion proteins of full length Pre S1 protein and GAL4 DNA binding domain presented transcriptional activation function in yeast. The transcription activating sequence was localized to the 21 to 47 amino acids of Pre S1 protein. Fusion proteins of full length Pre S1 and GAL4 DNA binding domain did not show transcriptional activation function in mammalian cells.
CONCLUSIONS: The transcription activating sequence of HBV Pre S1 protein in yeast overlaps the hepatocyte receptor binding site. The transcriptional activation function of HBV Pre S1 protein in yeast may prevent researchers from using yeast two-hybrid system to clone HBV receptor interacting with Pre S1 protein. However, the Pre S1 protein does not show transcriptional activation function in mammalian cells. Mammalian two-hybrid system may be a practical method to clone the HBV hepatocyte receptor interacting with Pre S1 protein.

OBJECTIVE: To investigate the expression and the role of iNOS expression in hepatic ischemia-reperfusion (I/R) injury.
METHODS: Male Wistar rats were subjected to 30-minute hepatic ischemia, then iNOS protein and iNOS mRNA expression in liver tissue was assessed by Western blot and RT-PCR analysis respectively at different time points after reperfusion. The effects of L-NAME (Nω-nitro-L-arginine methyl ester, a nonselective NOS inhibitor) or AE-ITU (aminoethytl-isothiourea, a relative selective inhibitor of iNOS) treatment were also evaluated.
RESULTS: High levels of iNOS protein and mRNA expression were detected in the liver tissue subjected to I/R, but not in the sham-operated rats. iNOS protein and iNOS mRNA expression reached a maximum on the first day after reperfusion and decreased later. The levels of iNOS protein and iNOS mRNA disappeared on 7th, 3rd day after reperfusion respectively. The high iNOS expression was correlated with hepatic dysfunction. L-NAME administration worsened hepatic dysfunction induced by hepatic I/R. In contrast, AE-ITU　administration showed mild protective effects against hepatic dysfunction induced by hepatic I/R.
CONCLUSION: Ischemia-reperfusion may induce or up-regulate the expression of iNOS protein and iNOS mRNA, which is detrimental to hepatic I/R injury.

OBJECTIVES: To construct a DNA vaccine capable of expressing the S gene of hepatitis B virus (HBV) and evaluate the expression of the recombinant S gene in vitro and in vivo.
METHODS: A cloned S-X gene fragment was inserted into an eukaryote expression vector to construct a recombinant expressing plasmid pCMV-SX. The recombinant plasmid was transcribed in vitro with a T7 promoter transcription system and transfected into a human hepatoblastoma cell line HepG2. The expression of the S gene was detected by Northern blot hybridization, Western blot hybridization, and enzyme-linked immunosorbent assay (ELISA), respectively. BALB/c mice were inoculated with the recombinant plasmid, and the efficiency of DNA-based immunization in eliciting anti-HBs was evaluated by ELISA.
RESULTS: In vitro transcription of the subcloned HBV S gene was confirmed by Northern blot hybridization. The results of Western blot hybridization and ELISA showed that the S gene was expressed exactly in HepG2. In immune experiment, 2 of 10 immunized mice were shown to induce antibody against HBsAg.
CONCLUSION: The recombination and expression of the S gene can be achieved successfully in vitro. And the recombinant plasmid is able to elicit humoral immune response in mice.

OBJECTIVE: To assess the effect of lipopolysaccharides (LPS) on the expression of CD14 and TLR4 in rat Kupffer cells (KCs).
METHODS: In rat KCs induced by LPS, the changes of CD14 and TLR4 expression were measured by RT-PCR and immunohistochemistry, and the expressions of TNF-αmRNA, IL-6mRNA or the concentrations of TNF-α, IL-6 were estimated by in situ hybridization, radioimmunoassay, and others.
RESULTS: The expressions of CD14 and TLR4 in KCs induced by LPS were markedly increased in a dose-dependent manner (10 mg/L-1 μg/L) or in a time-dependent manner (0.5 h-24 h), with the peaked expression of CD14 at 3-6 hours. The expressions of CD14 and TLR4 in KCs stimulated by the active mediators from KCs which had been exposed to LPS for 1 hour were obviously increased.
CONCLUSIONS: There is a close relationship between LPS or the active mediators from KCs induced by LPS and the expressions of CD14, TLR4. It is implied that the increase of TLR4, CD14 expression may be induced by LPS within 1-3 hours, and further increase of TLR4, CD14 expression may be correlated with the cytokines produced by KCs.

OBJECTIVE: To observe the distribution of copper in the subcellular structure for the understanding of primary pathogenesis of hepatolenticular degeneration (HLD).
METHODS: Skin fibroblasts taken from HLD patients were cultured as an in vitro model of HLD, and the control cells taken from healthy volunteers were clutured in the same way. The distribution of copper inside and outside of lysosomes in fibroblasts was detected by quantitative electron probe X-ray microanalysis. The relationship between the subcellular location of copper and the genotype of the patients, and relationship between the distribution of copper and the course of the disease were analyzed.
RESULTS: The content of Cu2+ inside lysosomes of HLD cells (14.6±2.1 mmol/kg) and of heterozygote cells (11.6±0.6 mmol/kg) was higher than that of normal cells (4.5±1.2 mmol/kg) (P<0.01). The content of Cu2+ outside lysosomes of HLD cells (17.5±4.2 mmol/kg) and of heterozygote cells (12.0±0.9 mmol/kg) was higher than that of normal cells (4.7±1.2 mmol/kg) (P<0.01). The distribution of copper in the subcellular structure was correlated with disease courses of HLD patients. With the progression of the disease, more copper was deposited in lysosomes (r=0.85, P<0.01). The content of copper in the diffused cytoplasmic compartment in HLD cells was correlated with that of sulfur (r=0.86, P<0.05), but not in heterozygote and normal cells.
CONCLUSIONS: In the early stage of HLD, copper is accumulated outside lysosome, which is paralleled with increase of metallothionein-like proteins (copper and sulfur-binding proteins). With the development of the disease, more copper is deposited inside lysosome than outside lysosome. We conclude that the up-regulation expression of copper and sulfur-binding proteins and copper accumulation in lysosomes may play an important role in lowering the ATP7B gene mutation-induced toxic effects of free copper on the cell.

OBJECTIVES: To detect the expression of human alpha-fetoprotein mRNA (AFPmRNA) in peripheral blood (PBL) of recurrent hepatocellular carcinoma (HCC) by using nested reverse transcriptase polymerase chain reaction (nested RT-PCR) and to discuss relations between AFPmRNA and recurrent HCC.
METHODS: Five ml peripheral blood samples were obtained from 19 patients with recurrent HCC. To identify HCC cells in PBL, liver-specific AFPmRNA was amplified from the total RNA extracted from the whole blood by nested RT-PCR.
RESULTS: Seven patients (36.8%) with recurrent HCC were detected AFPmRNA, and 12 patients with recurrent HCC were AFPmRNA negative (63.2%). There was no significant relation between patients with AFPmRNA positive and AFPmRNA negative (P>0.05) in judging the origin of recurrent HCC.
CONCLUSION: AFPmRNA may not be used as a marker of origin of recurrent HCC.

OBJECTIVE: To study the function of costimulation sign in tumor immunology and construct the new cell line B7+ Smmc7721.
METHODS: The B7 gene was transfected into the hepatocarcinoma cell Smmc7721 by liposma. Polymerase chain reaction (PCR) and reverse transcriptase-PCR (RT-PCR) were applied to test the result. MTT colorimetric assay was used to value the killing effect of lymphokine activated killer cells (LAK) activated by IL-2 on the transfected cell line and the original line.
RESULT: B7+ Smmc7721 was improved to be steadily expressed B7 molecule and LAK cells could more effectively act on the B7+ Smmc7721 cells.
CONCLUSION: The B7 gene can be transfected to hepatocarcinoma cells and can be expressed steadily in vitro, thus increasing the efficiency of LAK cells activated by IL-2 on them.

OBJECTIVE: To investigate the clinical epidemiology of intrahepatic cholelithiasis in Guangxi area, China.
METHODS: 8585 cases of cholelithiasis proved by surgery in a period of 19 years were analyzed retrospectively. Data were collected and analyzed by computer software package PEMS.
RESULTS: Cases of intrahepatic cholelithiasis accounted for more than one third of cases of cholelithiasis treated in the same period. The prevalence of intrahepatic cholelithiasis in farmers increased from 23.4% out of all cases with gallstone in 1981-1985 to 55.8% in 1991-1999. The constituent ratio of intrahepatic cholelithiasis in males was nearly the same in females. The peak prevalence age of patients with intrahepatic cholelithiasis ranged from 31 to 40 years, and the mortality was the highest among all bile stone cases.
CONCLUSION: Intrahepatic cholelithiasis is by no means a vanishing disease, especially in rural area.

OBJECTIVE: To study the correlation between human cholangiocarcinoma in the porta hepatis and the infection of hepatitis virus.
METHODS: Immunohistochemistry was used to detect HBxAg and HCV-C protein in formalin-fixed and paraffin-embedded samples taken from 68 patients with cholangiocarcinoma in the porta hepatis. The findings were reviewed against the clinical records of the patients.
RESULTS: Six patients (8.8%) were positive for HBxAg and 24 (35%) for HCV-C protein, respectively. One patient was positive for both HBxAg and HCV-C protein. There were statistically differences in the extent of differentiation, invasion, lymph-node metastasis, and treatment between the patients with cholangiocarcinomas in the porta hepatis with HB(C)V infection and those without infection.
CONCLUSIONS: HB(C)V infection is correlated to the development of cholangiocarcinoma in the porta hepatis. The tumor with HB(C)V infection may have a higher malignancy biologically and poorer prognosis. HBxAg and HCV-C protein may play an important role in the pathogenesis of cholangiocarcinoma in the porta hepatis.

OBJECTIVE: To develope a new enzyme immune assay (ELISA) for detection of M2 antibody specific for primary biliary cirrhosis (PBC) by using a triple hybrid clone as antigen, which coexpresses the three immunodominant lipoyl domains of PDC-E2, BCOADC-E2 and OGDC-E2 from human sources.
METHODS: After expressing autoantigens of PBC in prokaryote by constructing recombinant expressive plasmid successfully, the fusion protein was purified by affinity chromatography. The sera of 17 PBC patients were examined. As controls, the sera of 167 non-PBC patients and the sera of 1225 normal controls aged under 28 were examined.
RESULTS: None of the sera from the non-PBC patients or the normal controls was positive for anti-M2 shown by the new ELISA. However, the positivity rate for anti-M2 in the PBC patients was 100% (17/17), as shown by the new ELISA.
CONCLUSION: The detection system with a good sensitivity and specificity may be used as a powerful method for the diagnosis of PBC.

OBJECTIVE: To explore the mechanism of benign biliary stricture caused by bile duct trauma.
METHODS: A model of trauma of the common bile duct was established in 28 dogs and then repaired. The anastomotic tissues were taken on 3 days, 1 week, 3 weeks, 3 months, and 6 months respectively after operation and examined by using light microscopy and electromicroscopy. Macrophage, transforming growth factor beta 1 (TGF-β1) and α-smooth muscle actin (α-SMA) were studied immunohistochemically.
RESULTS: The mucosal epithelium of the common bile duct restored poorly, chronic inflammation lasted for a long time, fibroblasts proliferated actively, extracellular matrix overdeposited, and myofibroblasts functioned actively during the whole healing process. Immunohistochemical test showed a high expression of macrophage, TGF-β1 and α-SMA during the healing process lasting a long duration. Macrophages were found in the lamina propria under mucosa, TGF-β1 in the granular tissue, fibroblasts and endothelial cells of blood vessels, while α-SMA in the myofibroblasts and smooth muscle tissue.
CONCLUSIONS: The healing of the bile duct is in the mode of overhealing. Myofibroblast is the main cause for contracture of scar and stricture of the bile duct. The high expression of macrophage, TGF-β1 and α-SMA is closely related to active proliferation of fibroblasts, extracellular matrix overdeposition and scar contracture of the bile duct.

OBJECTIVE: To assess the short-term results of interventional therapy for malignant obstructive jaundice.
METHODS: In 82 patients with malignant obstructive jaundice, hepatocellular carcinoma was detected in 10 patients, carcinoma of gallbladder in 14, hilar biliary carcinoma in 22, pancreatic carcinoma in 20, and hilar metastatic carcinoma in 16. Percutaneous transhepatic biliary internal and/or external drainage (PTBIED) was performed in 61 patients and percutaneous transhepatic insertion of biliary stent (PTIBS) in 21.
RESULTS: The level of total serum bilirubin (TSB) was reduced in 71 patients and less markedly in others. The level of TSB of the 61 patients was reduced from 450.12±113.51 μmol/L before operation to 240.25±107.81 μmol/L and 90.91±101.72 μmol/L 1 and 2 weeks after operation respectively. The TSB level of the 21 patients was reduced from 410.53±98.13 μmol/L to 270.23±115.64 μmol/L and 105.43±97.85 μmol/L 1 and 2 weeks after operation, respectively. No significant difference was found in the effect between PTBIED and PTIBS. Short-term complications developed in 33 patients. Seven patients died 30 days after operation.
CONCLUSION: Interventional therapy may be simple, safe and effective in the treatment of malignant obstructive jaundice.

OBJECTIVE: To analyze factors predisposing to the infections associated with severe acute pancreatitis (SAP) and work out ways for their prevention.
METHODS: 208 patients with SAP treated at our hospital from January 1986 to December 2001 were retrospectively analyzed.
RESULTS: Statistical difference in the incidence of the infections was found between the following pairs: the groups of bloody or non-bloody ascites, paralytic ileus lasting shorter or longer than 5 days, Ranson’s scores lower or higher than 5, hematocrit lower or higher than 45%, CT Balthazar scores lower or higher than 7, and
between January 1986-June 1992 or July 1992-December 2001 admissions (χ2>7.58, P<0.05), while no statistical difference established between the groups of biliogenic and non-biliogenic pancreatitis, serum amylase <200 U/L and ≥200 U/L, serum calcium <2 mmol/L and ≥2 mmol/L or groups of total parenteral nutrition shorter or longer than 7 days (χ2<1.61, P>0.05).
CONCLUSIONS: Occurrence of infection in patients with SAP is closely related with bloody ascites, paralytic ileus (≥5 days), Ranson’s scores (≥5), hematocrit (≥45%) and CT Balthazar scores (≥7), but not with pathogenesis, serum calium or total parenteral nutrition. Comprehensive prevention of pancreatic infection and individualized therapy may reduce the incidence of infection.

OBJECTIVE: To investigate the effect of FasL gene transfer to islet cells on pancreatic islet allografts.
METHODS: A recombinant and replication-deficient type-5 adenovirus encoding murine FasL (AdV- FasL) was constructed by the method of calcium phosphate precipitation. Pancreatic islets were infected with the recombinant adenovirus AdV-FasL, and transplanted into diabetic recipients. FasL expression was detected by RT-PCR and immunohistochemistry. The survival of allografts and the apoptosis of gene transferred islet allografts were analyzed.
RESULTS: All animals receiving islet allograft alone returned to a diabetic state by several days (mean survival time 6.3±0.6 days). Compared with the control group, no delayed rejection and prolonged survival of allografts were observed in the group of FasL gene transfer. The rejection was accelerated and the allograft survival was shortened to 3.4±0.2 days (P<0.05). Pancreatic islets infected with AdV- FasL demonstrated positive staining of FasL at 24 h, with an increased intensity at 48 h, but not in AdV-5 infected or uninfected islets. TUNEL labeling of pancreatic islet allografts at 24, 48 h revealed apoptosis that was not in AdV-5 infected allografts.
CONCLUSIONS: Though co-transplantation of FasL-expressing testicular cells can induce privilege of islet allografts and prolong allograft survival, direct expression of FasL on islet allografts infected with AdV-FasL may accelerate islets rejection by islet apoptosis and granulocyte infiltration.

OBJECTIVE: To assess the diagnostic value of endoscopic pancreatic duct brushing in detecting mutation of the K-ras gene at codon 12 in cytologic specimens from patients with pancreatic cancer.
METHODS: Thirty-five patients treated at Changhai Hospital, Shanghai between 1999 and 2001 were enrolled. Their cells obtained by pancreatic duct brushing during endoscopic retrograde cholangiopancreatography (ERCP) were suspended with phosphate buffer solution (PBS). DNA of the cells was extracted and mutation of the K-ras gene at codon 12 detected by means of PCR-SSCP.
RESULTS: The K-ras gene mutation rate of pancreatic cancer was 70%, which was higher than that of chronic pancreatitis (14%, P<0.05). K-ras gene mutation was not found in patients with pancreatic cystocarcinoma and duodenum carcinoma. As to the location of pancreatic cancer, no significant difference was observed between the head, the body and tail. The sensitivity, specificity, accuracy of pancreatic duct brushing in detecting pancreatic cancer was 70%, 94%, and 83%, respectively.
CONCLUSION: K-ras analysis of pancreatic brushing samples is helpful in the diagnosis of patients with early pancreatic cancer.