Abstract:Abstract:Objective:To summarize the clinical experience of liver transplantation in the treatment of hepatocellular carcinoma (HCC).
Methods :From June 2004 to March 2007，70 consecutive HCC patients underwent liver transplantation in our hospital, including classic orthotopic liver transplantation in 41 cases, and piggyback liver transplantation in 29 cases. All data of patients were retrospectively analyzed.
Results:All liver transplantation were successfully conducted .The average warm ischemia duration was 4.5 minutes, and cold ischemia duration was 8 hours. There were 3 cases of postoperative deaths. Surgical complications were intra-abdominal hemorrhage in 2 cases， and biliary anastomotic stricture in 4 cases. Sixty-seven cases were followed up for 12-33 (average 21) months, 10 cases had recurrence of liver cancer after transplatation and 1 dead.
Conclusions:Liver transplatation can used on selected patients with HCC. Successful liver transplantation relies on good quality of liver graft， and idealized technique of vascular and bile duct reconstruction are key factors of liver transplantation. Proper postoperative management can effectively reduce the complications of operation.

Abstract:Abstract:Objective:To investigate the strategy and outcome of prevention of hepatitis B virus(HBV) reinfection after orthotopic liver transplantation (OLT).
Methods :A total of 120 patients with chronic fulminant hepatitis B，end-stage of liver cirrhosis and liver carcinoma were given hepatitis B immunoglobulin (HBIG) plus lamivudine or adefovir after OLT to prevent HBV reinfection．The serum HBV and HBV DNA were determined and clinical observation were performed.
Results:Follow-up time ranged from 23-59 (median 45.2) months. Four patients had reinfection with positivity of HBsAg，HBeAb and HBcAb, the serum HBV DNA was also positive．All 4 patients became HBV-DNA negative after treatment and folloed-up for 19-26 months, but HBsAg remained positive.
Conclusions:HBIG plus lamivudine or adefovir can effectively prevent HBV reinfection after OLT. Load level of HBV-DNA before OLT was related with HBV reinfection after operation.

Abstract:Abstract:Objective:To explore the effects of Gln-glutamine (Gln-) supplemented TPN and traditional TPN on immunofunction, incidence of both rate of acute rejection and infection, and protein synthesis function of transplantation liver.
Methods :Fifty patients with liver transplantation were randomly divided into two groups: group TPN without Gln- and group TPN with Gln-. Patients received isocaloric ［104.6 kJ /(kg·d)］ and isonitrogenous ［(0.16g/(kg·d)］ TPN for seven days. IgG，IgA，IgM, CD3,CD4/CD8, prealbumin(PAB) were monitored on the second and the ninth day after transplantation.
Results:IgG, IgA, CD3, CD4 /CD8,PA levels of the group TPN with Gln- had significant elevation on the ninth day as compared to those on the second day after operation. The incidence rate of infection of the group TPN with Gln- was significantly lower than the group TPN without Gln-, while the incidence rate of acute rejection had no significant difference between the two groups.
Conclusions:Gln-glutamine supplemented TPN can enhance the immunofunction of patients after liver transplantation, stimulate protein synthesis function and lower the incidence rate of infection, and does not increase the incidence of acute rejection.

Abstract:Abstract:Objective:To observe the manifestations and judgement methods of acute rejection reaction of orthotopic liver transplantation in Wistar-SD rats.
Methods :The survival conditions after operation in rats was observed, and liver function tests and histopathology examination were used to study the manifestations of acute rejection reaction of orthotopic liver transplantation in Wistar-SD rats.
Results:Wistar-SD experiment group had moderate to severe acute rejection reaction after OLT. Alanine aminotransferase, aspartate aminotransferase, total bilirubin in blood serum after orthotopic liver transplantation in rats was obviously elevated on 1-3 d and 7 d after OLT, which are significantly different from every corresponding time point of control group(P<0.05).
Conclusions:Liver transplantation from donor Wistar rats to recipient SD rats can result in moderate to severe acute rejection reaction, and changes in results of liver functional parameters can indicate whether the acute rejection reaction has occurred.

Abstract:Abstract:Objective:To investigate the effects of the ischemic postconditioning (IPO) on the ischemia reperfusion injury (IRI) in rat livers.
Methods :One hundred and tweenty Wistar rats were randomly divided into ischemia reperfusion group (IRI), ischemic postconditioning (IPO) group and sham operation (S) group. Liver tissues were harvested at 0, 0.5, 1, 2, 4, 8, 12 h and 24 h after reperfusion in both IPO and IRI groups. The mRNA expressions of the immediate early gene c-fos and c-jun were detected with RT-PCR methods.
Results:(1) The expression of c-fos and c-jun both maintained a high level from 0.5 h to 2 h after reperfusion in IRI group, and the peak time occurred at 1 h after reperfusion. After 4 hours of reperfusion, the c-fos expression began to decrease gradually while the c-jun expression still remained high.(2) At every time point, the c-fos expressions in IPO group were lower than IRI group (P>0.05).(3) Compared with IRI group, the c-jun expression at 0.5, 1 h and 2 h after reperfusion in IPO group was significantly lower (P<0.05).
Conclusions:IPO can impact the immediate early gene expression at the very early moment after reperfusion, which may be part of the mechanisms of IPO protection effect on ischemia reperfusion injury.

Abstract:Abstract:Objective:To explore the selection of indicator to judge the severity of hepatic ischemic and reperfusion injury(IRI).
Methods :Local warm hepatic IRI model was established routinely in rat. The serum alanine aminotransferase(ALT) levels were determined and specimens were sent for pathologlical test at time of 0, 1, 2, 4, 6, 12, 24, 48 h of reperfusion, after ischemic time of 60 and 90 min respectively. The rats were re-grouped into 4 groups of different severity according to the grading systems for IRI used in our institute， and the relationship between ALT level and pathological changes was analyzed.
Results:After 60 min of ischemia time, there was mild IRI in 75%, intermediate IRI in 14.6% and severe IRI in 10.4% of rats, respectively. After 90 min of ischemia time, there was mild IRI in 41.7%, intermediate IRI in 25%,severe IRI in 16.7%, very severe IRI in 16.7% of rats respectively. The results showed that the more severe the degree of IRI, the higher rate of the cell death, and the higher of the ALT value.
Conclusions:The serum ALT level can directly reflect the severity of IRI. The more severe the degree of IRI, the higher was the ALT level, and the higher was the cell death rate.

Abstract:Abstract:Objective:To explore the protective effect of portal vein administration of propofol on hepatic ischemia reperfusion injury (HIRI) and its mechanism.
Methods :Thirty-two male rabbits were randomly allocated into four groups:Group A (sham operation group), the abdomen was only opened and closed;group B,the hepatic inflow was occluded for 30 min,and reperfused for 60 min; group C,the same managment as group B + propofol injected through jugular vein;group D,the treatment same as group B + propofol injected through portal vein.Drug injection was completed 20 min before hepatic inflow occlusion. Serum ALT and AST, and endothelin-1 (ET-1) and nitric oxide(NO)in the hepatic tissue and blood, and the content of ATP in hepatic tissue were determined.
Results:The level of ET-1 in plasma and hepatic tissue was significantly increased in group B compared to group C and D (P＜0.01、P＜0.05). The level of NO in serum and hepatic tissue in group B was significantly lower than that in group C,D (P＜0.01). The content of ATP in hepatic tissue was significantly decreased in group B compared to group C and D (P＜0.01).Comparing the efficiency of administration routes, propofol delivered through portal vein have better protective effect on HIRI.
Conclusions:Propofol delivered through portal vein has significant protective effect on HIRI. Propofol may adjust disbalance between ET-1 and NO to improve microcirculation,and elevate the content of ATP in the hepatic tissue.All of these contribute to protective effect on HIRI.

Abstract:Abstract:Objective:To study the protective effect and mechanism of safflower solution on warm ischemia reperfusion injury (IRI) of liver in rats.
Methods :Male Sprague-Dawley rats were randomly divided as 4 groups: Group S (sham group); group I/R (IRI group); group IPC (ischemic preconditioning group); group SPC (safflower solution preconditioning group). The rats were sacrificed at 24 h after reperfusion, serum levels of ALT and AST were measured and HE staining of liver tissues were made to detect rat liver histological changes and grade liver IRI(Suzuki score); apoptosis were monitored by TUNEL; TNF-α, MIP-2 and ICAM-1 mRNA of liver were measured by RT-PCR; NF-κB gene products of liver were detected by Western blotting.The mean value of all the assay tests was compared.
Results:After 24 h reperfusion, compared with group I/R, the liver function (ALT and AST) in group SPC and group IPC was significantly improved (P＜0.05), Suzuki scores was significantly decreased (P＜0.05), the level of hepatic TNF-α, MIP-2 and ICAM-1mRNA was significantly decreased (P＜0.05), and the expression of hepatic NF-κB protein was also decreased (P＜0.05).
Conclusions:Donor pretreatment with safflower solution can decrease the severity of warm IRI of liver graft by downregulation of proinflammatory cytokines TNF-α, MIP-2 and ICAM-1 mRNA and by its antiapoptotic effects.

Abstract:Abstract:Objective:To study the role of 1, 4, 5-trisphosphate inositol ( IP3) and bax gene expression in inhibiting transplanted hepatocellular carcinoma of nude mice by quercetin.
Methods :After animals with hepatocellular carcinoma were treated with quercetin 1mg/kg/d (ip)for 3 weeks, the volume and weight of tumor was measured, and IP3, Bax mRNA, and Bax protein were assayed by IP3-［3H］ Birtrak assay,RT-PCR, and Western blotting, respectively; and compard with coutrol group.
Results:The tumor volume and weight of animals treated with quercetin were lower than those of control［(15.8±10.1) mm3 vs. (52.3±26.5 mm3; (44.8±10.4) mg vs. (91.3±31.4) mg］，IP3 content was lower than that of control［（15.9±2.8）pmol/mg protein vs （35.3±6.6）pmol/mg protein］，Bax mRNA expression was not significantly different between the 2 groups［RI which was the gray degree multiply area of bax / the gray degree multiply area of β-actin (0.64±0.12) vs. (0.56±0.15)］，Bax mRNA expression was higher in group treated with quercetin than that of control［(3.16±0.95) vs. (1.37±0.48)］.
Conclusions:Quercetin can inhibit growth of transplanted hepatocellular carcinoma of nude mice by reducing IP3 production and bax protein expression.

Abstract:Abstract:Objective:To study the effect and mechanism of adenovirus-mediated mda-7/IL-24 and/or adriamycin treatment of hepatoma in nude mice.
Methods :The recombinant adenovirus vector carrying melanoma differentiation-associated gene-7 (Ad-mda-7) and/or ADM were injected into the tumor-bearing mice. Their effects on the volume of the tumor and the survival rate of the mice were observed, and the apoptosis of cancer tissue was detected by TUNEL staining in each group.
Results:Ad-mda-7 was successfully constructed and expressed in vivo. Compared with the other three groups, the mice treated with Ad-mda-7 combined with adriamycin had longer average survival time (83.8±4.82) d (P<0.01); the average size of tumor in combined group was reduced significantly, the tumor inhibition rate was 79.78%, which was higher than the other groups (P<0.05). Enhanced induction of apoptosis by Ad-mda-7 combined with adriamycin was observed. The TUNEL-positive cells showed that apoptotic index in combination therapy group was 38.1%±4.2 %，which was higher than that of the other 3 groups (P<0.05).
Conclusions:Ad-mda-7 combined with adriamycin has stronger antitumor potency and has more synergistic effect on metastastic hepatoma mouse model than mda-7/IL-24 or adriamycin alone, and the mechanism is related to the promotion of apoptosis of tumor cells.

Abstract:Abstract:Objective:To investigate the effect of mitofusin-2 gene (mfn2) on apoptosis of hepatocellular carcinoma cell strain HepG2 and its possible mechanism.
Methods :Plasmid pEGFPmfn2 was transfected into hepatocellular carcinoma cell strain HepG2 by Lipofectamine 2000 in vitro. The mRNA of Mfn2 expression was detected by RT-PCR and protein expression by Western-blot. The cell proliferation of HepG2 was measured by MTT assay and cell apoptosis was observed in Annexin-V/PI by flow cytometry. The ultramicrostructure of cell was observed by electron microscope. The expression of apoptosis related genes was observed by RT-PCR.
Results:Mitofusin-2 gene was expressed in transfected HepG2/mfn2 stably and it could obviously inhibit cell proliferation by MTT assay. Exogenous mfn2 gene significantly induced cell apoptosis and induced the break down, disappearance and groundsubstance rarefaction of chondrosome cristae. Expression of Bcl-2 and survivin genes were reduced in HepG2/mfn2.
Conclusions:Exogenous mfn2 could inhibit proliferation of HepG2 and induce its apoptosis. The downregulation of apoptosis related genes may be the mechanism for inducing apoptosis.

Abstract:Abstract:Objective:To analyze the expression of HLJ1 in the tissues of hepatocellular carcinoma(HCC) and the correlation between its expression and clinical pathological characteristic.
Methods :HLJ1 mRNA expression level was determined in 32 HCC tissues and their associated noncancerous liver tissues by reverse transcription polymerase chain reaction (RT-PCR). The correlation of HLJ1 expression level with clinicopathologic variables was also analyzed.
Results:HLJ1 mRNA expression level was significantly lower in HCC than in associated noncancerous liver tissue ［(1.18±0.82) vs. (1.92±1.15), P＜0.05］. Importantly, the HLJ1 mRNA expression level correlated with the Edmondson-Steiner grade (P=0.010) and vein invasion (P=0.001).
Conclusions:Our study indicates that HLJ1 mRNA expression level is significantly lower in HCC tissues than in associated noncancerous liver tissue. HLJ1 may participate in the cell proliferation and invasiveness of HCC cell.

Abstract:Abstract:Objective:To investigate the effect of hepatitis c virus NS3 gene on the expression of phosphorylate tyrosine proteins in human hepatocyte.
Methods :HCV NS3 plasmids and pRcCMV plasmids were stably transfected into QSG7701 cells, which were named as pRcHCNS3/QSG cells and pRcCMV/QSG cells respectively. Western Blot and immunocytochemistry was used to detect the expression of phospho-p44/42, phospho-p38 and phospho-SAPK/JNK in the pRcHCNS3/QSG cells, pRcCMV/QSG cells and QSG7701 cells, respectively. Immunoprecipitation analysis was employed to investigate the effect of HCV NS3 on the phosphotyrosine protein expression.
Results:The expressions of phospho-p44/42, phospho-p38 and phospho-SAPK/JNK were found in pRcHCNS3/QSG cells, pRcCMV/QSG cells and QSG7701 cells. The positive signals of phospho-p44/42, phospho-p38 and phospho-SAPK/JNK protein were localized in the nucleus of cells. The signals intensity of the pRcHCNS3/QSG cells was stronger than the other the two cells. The expression of phosphotyrosine protein was also up-regulated in pRcHCNS3/QSG cells.
Conclusions:HCV NS3 may affect cell signaling pathway through activating tyrosine kinase and followed with activation and abnormal phosphorylation of several cytoplasmic signaling molecules such as MAPKs, and induce unlimited hepatocyte transformation and tumor development.

Abstract:Abstract:Objective:To study the expression of the adenoviral vector containing myr-HA-Akt gene in the liver of cirrhotic rats in vivo.
Methods :myr-HA-Akt cDNA obtained from the plasmid pcDNA3.1-myr-HA-Akt was cloned into the plasmid pDC316. Then, pDC316-myr-HA-Akt was cotransferred with adenoviral backbone vector into 293 cells. The recombinant adenovirus was reproduced and purified. The recombinant adenovirus was identified by the cytopathic effect of 293 cells, polymerase chain reaction(PCR),and gene sequencing for myr-HA-Akt cDNA. TCID50 assay was performed to determine the titer of virus. After the adenovirus infected the hepatic cirrhosis rat models via the tail vein, protein and phosphorylation status of Akt were examined by Western blotting.
Results:Infected 293 cells showed significant cytopathic effect. Products of PCR confirmed the presence of recombinant adenovirus. The identification result by DNA sequence analysis showed that myr-HA-Akt cDNA was cloned to pDC316 correctly and homologously recombinated with pBHGloxΔE1, and 3Cre and Ad-myr-HA-Akts were packaged successfully. The titer of virus was 5.5×1011 vp/ml. The expression of Akt in the liver of cirrhotic rats was verified by Western blotting.
Conclusions:The recombinant adenoviral vector containing myr-HA-Akt was constructed and the transgene of hepatic cirrhosis rats expressed Akt gene in vivo successfully. It provides a basis for the further study of treatment for hepatic cirrhosis by Akt gene.

Abstract:Abstract:Objective:To explore the variations of regeneration hormones after hepatectomy for liver cancer, and evaluate the relationship between the liver regeneration hormones and cancer recurrence.
Methods :The clinical data of 129 patients with primary hepatic carcinoma in our hospital from Dec 2004 to Dec 2005 were collected. The patients were divided into three groups according to their recurrent times, which were one-month recurrence group, 6 months recurrence group and one-year recurrence group. And at the same time, 40 cases of liver cancer that received TAE treatment were as contrast group. Serum HGF value was detected before operation and 1, 3, 7,10 and 14 days after operation. c-met, which is the receptor of HGF, was also detected as c-met mRNA and protein expression in cancer tissue and near-carcinoma liver tissue by semi-quantitative RT-PCR and Western Blot. The differences between the level of expression and the time of recurrence were compared, and the results were also compared with pathological indexes.
Results:Serum HGF value elevated after hepatectomy, the crest time appeared at about 10 days after the operation, and decreased after 14 days. The elevated values of HGF in large HCC tumors were markedly higher than those in small HCC tumors. The change of c-met mRNA and protein levels, revealed that the earlier the recurrence in both large and small HCC, the higher the c-met levels, and the higher the rate of vascular cancer emboli.
Conclusions:There is marked elevation of HGF level after hepatectomy in patients with liver carcinoma, and the over expression of c-met of the tumor may be related to its early postoperative recurrence.

Abstract:Abstract:Objective:To explore the clinical value of B ultrasound-guided psychro-circulation percutaneous microwave coagulation therapy(PMCT) in the treatment of liver cancer.
Methods :One hundred and sixteen liver cancer patients with 154 lesions were selected to receive psychro-circulation PMCT in our department from July 2004 to December 2007. The pre-and postoperative alpha fetoprotein(AFP) levels were detected. The diameters and blood flow state of the tumors were measured and compared by use of B ultrasound and computerized tomographic scanning(CT).The postoperative 1,2 and 3 y survival rate of those patients was followed up and recorded.
Results:No mortality or severe complications occurred in this series.Preoperative serum AFP level of the patients was (421.6±36.5)μg/L and decreased obviously to （232.5±25.6）μg/L at 1 month after psychro-circulation PMCT（P<0.01）.Preoperative tumor diameter was (48.6±10.2) mm and diminished to (37.5±7.8) mm at 1 month postoperatively（P<0.05）.At 3 months postoperatively,the liver tumors disappeared completely in 15 cases, obviously decreased in size in 86 cases and no marked changes occurred in 15 cases. In the patients with primary hepatic carcinoma,the postoperative 1,2 and 3 y survival rate was 89.4%,67.1%, and 43.8%,respectively. In the patients with secondary hepatic carcinoma,the postoperative 1,2 and 3 y survival rate was 82.1%,53.8%, and 24.0%,respectively.
Conclusions:Psychro-circulation PMCT is an effective method for treating liver cancer and is especially suitable for inoperable patients, because of its mini-trauma,few complications and convenient performance.

Abstract:Abstract:Objective:To investigate the possible mechanism of pancreaticobiliary maljunction (PBM) on development of congenital choledochal cyst (CCC).
Methods :Tweenty-one patients with choledochal cysts initially diagnosed by ultrasonography, then underwent ERCP. The concentrations of amylase (AMS) and C-reactive protein (CRP) in bile removed through selective biliary catheterization was analyzed.
Results:Each of 21 cases of PBM (including 13 cases of C-P type and 8 cases of P-C type) had significantly higher concentrations of AMS and CRP compared with control group (P<0.01). The elevations of AMS and CRP in C-P type were much higher than those in P-C type. (P<0.05).
Conclusions: PBM was likely the anatomic foundation of CCC formation. However, the activated, highly concentrated pancreatic enzymes resulting from pancreatic juice reflux into the bile ducts can cause injury of the walls of bile ducts, and trigger CCC formation.

Abstract:Abstract:Objective: To study the clinical characteristics and essential points of diagnosis and treatment of hepatolithiasis associated with cholangiocarcinoma.
Methods :The clinical data of 55 cases of hepatolithiasis associated with cholangiocarcinoia were analyzed retrospecively. Among them, radical resection was performed in 23 cases and palliative resetion in 32 cases.〖JP3〗
Results:The pathological diagnosis was biliary adeniocarcinoma〖JP〗 in 41 cases, and mucinous carcinoma in 14 cases. Fifty-one patients were followed up for an average 3 years and 9 months, the radical treated group survived an average of 26 (13-45) months, and the palliative conservative treatment group survived an average of 10 (5-14) months. The 1,2,3-year survival rates of radical treatment group was 96.4%, 54.6%, and 24.5% respectively, and for palliative treatment group, the 1-year survival rate of 37.8%, and none survived for 2 years.
Conclusions:Hepatobiliary gallstones can repeatedly develop inflammation that can lead to cholangiocarcinoma. Radical resection can prolong survival time, while palliative surgical treatment can improve quality of life. The key to improve the therapeutic outcome of hepatolithcasis with cholangiocarcinoma is to strive for early complete treatment of hepatobiliary gallstone.

Abstract:肝缺血再灌注损伤（HIRI）发生机制极为复杂，是涉及多因素、多脏器的全身性疾病。笔者就肝移植过程中（HIRI）发生胰腺缺血再灌注损伤(PIRI)的相互联系和作用机制等相关研究的最新进展作一综述。