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Objective: To evalutate the safety and efficacy of infrahepatic inferior vena cava clamping robot-assisted laparoscopic liver resection. Methods: All data about 24 patients with robotic liver resection at Hepatic Surgery Center,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology between February 2015 and December 2017 were collected and analyzed. These patients were divided into two groups based on different methods to decrease central venous pressure. Eight patients(6 males and 2 females,aged 49 years(range:50 to 56 years)) were applied with infrahepatic inferior vena cava clamping,and the other 16 matched cases (15 males and 1 female,aged 53 years(range:38 to 69 years)) were categorized into lowering central venous pressure group. Intraoperative blood loss,blood transfusion,intraoperative hemodynamic parameters,postoperative complications,and renal function were compared by t -test,non-parametric test,χ 2 test,or Fisher exact test. Results: There was significantly difference in the intraoperative blood loss between the infrahepatic vena cava clamping group and the lowering central venous group(200(220) ml (range:100 to 400 ml) vs. 750(800) ml (range:100 to 2 000 ml), Z =‒2.169, P =0.030). The clamping time of portal triad and infrahepatic inferior vena cava were 24 (18) minutes and 29 (20) minutes in the infrahepatic inferior vena cava clamping group, and portal triad clamping time was 23 (23) minutes in the low central venous group. There was no significant difference between the two groups ( Z =‒0.323, P =0.747). There was no intraoperative blood transfusion in the infrahepatic inferior vena cava clamping group, and 5 cases in the low central venous group, with a transfusion volume of 1.5(1.5)U. The difference between the two groups was statistically significant ( Z =‒3.353, P =0.001). However, the mean arterial pressure in the infrahepatic vena cava clamping group decreased from (88.6±4.9) mmHg to (67.4±3.8) mmHg(1 mmHg=0.133 kPa), which was lower than that of lowering central venous group (72.4±3.3) mmHg ( t =2.315, P =0.003). And there were no significant differences related to postoperative complications rate or hepatic and renal function in both groups. Conclusion: The infrahepatic inferior vena cava technology is safe and feasible to decrease central venous pressure during robotic liver resections,which will not affect the recovery of hepatic and renal functions.

The 2019 coronavirus disease(COVID-19) is a highly infectious disease, has a long incubation period and a variety of clinical manifestations, which has a significant impact on public health and life. Afterwards, scientific and standardized work processing during the epidemic is of great significance for prevention and control. In order to implement the central government's decision-making deployment and defeat the COVID-19 as soon as possible, we had focused on the key points in the clinical work of general surgery according to latest relevant guidelines, literature and experience in epidemic prevention. Finally, we drafted the prevention and control strategies and recommendations to make a reference for medical staff of general surgery to fight against COVID-19.

Novel coronavirus pneumonia (NCP) is a highly infectious disease, has a long incubation period and a variety of clinical manifestations, which has a significant impact on public health and life. Afterwards, scientific and standardized work processing during the epidemic is of great significance for prevention and control. In order to implement the central government's decision-making deployment and defeat the NCP as soon as possible, we had focused on the key points in the clinical work of general surgery according to latest relevant guidelines, literature and experience in epidemic prevention. Finally, we drafted the prevention and control strategies and recommendations to make a reference for medical staff of general surgery to fight NCP.

Objective: To investigate the safety and feasibility of longitudinal transpancreatic U-sutures invaginated pancreatojejunostomy (Chen's pancreaticojejunostomy technique) in laparoscopic pancreaticoduodenectomy (LPD). Methods: Clinical data of 116 consecutive patients who underwent LPD using Chen's pancreaticojejunostomy technique in Hunan Provincial People's Hospital from May 2017 to December 2018 were retrospectively analyzed. Among these patients, 66 were males and 50 were females. The median age was 58 years old (32-84 yeas old). All 116 patients underwent pure laparoscopic whipple procedure with Child reconstruction method, using Chen's pancreaticojejunostomy technique. The intraoperative and postoperative data of patients were analyzed. Results: All 116 patients underwent LPD successfully. The mean operative time was (260.3±33.5) minutes (200-620 minutes). The mean time of pancreaticojejunostomy was (18.2±7.6) minutes (14-35 minutes). The mean time of hepaticojejunostomy was (14.6±6.3) minutes (10-25 minutes). The mean time of gastrojejunostomy was (12.0±5.5) minutes (8-20 minutes). The mean estimated blood loss was (106.0±87.6) ml (20-800 ml). Postoperative complications were: 11.2%(13/116) of cases had postoperative pancreatic fistula (POPF), including 10.3% (12/116) of biochemical fistula and 0.9%(1/116) of grade B POPF, no grade C POPF occurred; 10.3%(12/116) had gastrojejunal anastomotic bleeding; 3.4%(4/116) had hepaticojejunal anastomotic fistula; 3.4%(4/116) had delayed gastric emptying; 4.3% (5/116) had localized abdominal infection; 12.1%(14/116) had pulmonary infection; postoperative mortality were 0(0/116) and 1.7%(2/116) within 30 days and 90 days, respectively. One patient died of massive abdominal bleeding secondary to Gastroduodenal artery pseudoaneurysm rupture, the other patient died of extensive tumor recurrence and metastasis after surgery. Conclusions: Chen's pancreaticojejunostomy technique is safe and feasible for LPD.It is an option especially for surgeons who have not completed the learning curve of LPD.

In the past 20 years,the advancement and breakthrough of applied basic research,the invention and renewal of medical devices,the popularization and improvement of minimally invasive techniques,the development and debate of innovative surgical methods and the establishment of the diagnosis and treatment standard brought the unprecedented development momentum to traditional liver surgery.This article combined the domestic and foreign related literature and the research results of our team. The current status and progress of the application of preoperative evaluation and surgical planning of liver surgery,surgical techniques of liver resection,laparoscopic and robotic techniques in liver surgery,enhanced recovery after surgery and associating liver partition and portal vein ligation for staged hepatectomy are briefly discussed.

Objective: To explore the clinical effect and safety of minor liver resection for hilar cholangiocarcinoma (HC) of Bismuth-Corlette type Ⅲ and Ⅳ. Methods: From May 2007 to May 2017, the clinical data of 108 patients with Bismuth-Corlette type Ⅲ and Ⅳ HC underwent hepatectomy were collected and analyzed retrospectively.There were 56 males and 52 females, aged (57.2±5.3) years (ranged 48-76 years) .Among the 108 cases, there were 51 cases of type Ⅲa, 40 cases of type Ⅲb and 17 cases of type Ⅳ. Small-scale hepatectomy (≤3 hepatectomy) was performed in 70 cases, including 8 cases of 4b segment resection, 28 cases of 4b segment+5 segment resection, and 34 cases of partial 4 segment+partial 7 segment+partial 1 segment resection. Large-scale hepatectomy was performed in 38 cases (>3 segments) , of which 30 cases were treated with 2 segments+3 segments+4 segments+1 segment, and 8 cases were treated with 5 segments+7 segments+8 segments+1 segment. t' test was used to analyze the data which did not conform to the normal distribution, and χ(2) test was used to calculate the incidence of postoperative complications and the 1, 3, and 5-year cumulative overall survival rate. Results: (1) The operation time of minor liver resection group ((180±25)minutes) was shorter than that of major liver resection group ((210±35)minutes) ( t' =4.676, P< 0.05) , the amount of blooding operation time of minor liver resection group ((310±80)ml) was less than that of major liver resection group ((500±110)ml)in the operation ( t' =9.385, P< 0.05) , and the difference was statistically significant. (2) The incidence of complications was lower in minor liver resection group and major liver resection group, and the difference was statistically significant (χ(2)=5.230, P< 0.05) . (3) The actual 1-, 3- and 5-year survival rates were 87.1%, 58.4%, 30.0% and 84.2%, 57.9%,31.6%, respectively. There were no significant differences in survival rates in two groups in 1-, 3- and 5-year survival rates (χ(2)=0.177, P= 0.674; χ(2)=0.005, P= 0.946; χ(2)=0.029, P= 0.865) . Conclusions: Compared to patients with major liver resection, Minor liver resection for selected patients with HC of Bismuth-Corlette Ⅲ and Ⅳaccording to our criteria achieved better long-term outcomes. Chen's biliojejunostomy is a simple, effective and safe method, which can be widely used when there are multiple biliary intestinal anastomosese.

Liver resection is the mainstay of treatment for patients with hepatocellular carcinoma (HCC). Most of HCC patients are associated with varied degrees of liver cirrhosis.Severity of liver cirrhosis adversely affects the outcomes of liver resection, and also plays a vital role in making an appropriate surgical strategy for HCC.In current surgical practice for HCC, liver function and functional reserve are the focus of preoperative evaluation. Liver cirrhosis is still widely regarded as an one-stage entity. The pathological severity of liver cirrhosis is largely ignored. As neither liver function nor functional reserve can reflect the pathological severity of liver cirrhosis when liver function is at the stage of compensation. Preoperative evaluation on the severity of cirrhosis has not been established in a surgical setting.Thus, there is an urgent need to stage the severity of cirrhosis in surgical practice in order to make more precise surgical modalities for individual patients.This article mainly introduces the ongoing research progress in staging the severity of liver cirrhosis while treating HCC at Hepatic Surgery Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, and emphasizes the importance of staging the severity of cirrhosis in surgical treatment of HCC.

The surgical management of liver cancer has evolved rapidly.With the application of new technologies, such as regional ablation, laparoscopic and robotic surgery, surgery indications have been broadened. Based on the relevant guidelines for hepatocellular carcinoma published abroad in recent years, this paper gives interpretation of"Expert consensus on the option of surgical management of hepatocelluar carcinoma" made by Liver Surgery Group, Surgery Branch of Chinese Medical Association.

Objective: To explore the effectiveness of three-dimentional(3D)reconstruction technique in safety assessment of hepatectomy for large hepatocellular carcinoma(HCC)., Methods: The clinical records of 28 patients who underwent resection of HCC greater than 10 cm in diameter from January 2013 to December 2015 at Department of Hepatic Surgery, Tongji Hospital, Tongji Medical College Huazhong University of Science and Technology were studied retrospectively. All patients underwent enhanced computer tomography (CT), then 3D images of liver and tumor were reconstructed by uploading the CT imaging data to IQQA-Liver system. The individual surgery plan was simulated and liver volume (LV), standard liver volume(LV), tumor volume(TV), functional liver volume(FLV), excised liver volume(ELV), excised functional liver volume (EFLV), residual functional liver volume (RELV) were calculated. Simulated surgery plans were compared with actual procedures. ELV was compared with actual excised liver volume (AELV) by paired Student's t test. Postoperative complications and motility were analyzed. The correlation between TV and EFLV, RFLV, RFLV/FLV, RFLV/SLV were calculated by Spearman test., Results: TV ranged from 202 cm(3) to 2 125 cm(3,) RELV ranged from 401 cm(3) to 1 633 cm(3).There were 13 patients whose RFLV/LV<30% and 28 patients whose RFLV/FLV>30%(34.8%-94.0%). RFLV/SLV ranged from 35.9% to 139.0%.All simulated surgery plans matched with the actual operation procedure. ELV was equal to AELV, which confirmed by the high precision of IQQA-Liver system(t=0.636, P>0.05). No severe complications (hepatic encephalopathy or liver failure) and perioperative death occurred after operation. Positive correlation was observed between TV and RFLV, TV and RFLV/FLV, TV and RFLV/SLV(r=0.641, 0.629 and 0.732, all P<0.01). Negative correlation was observed between TV and EFLV (r=-0.539, P<0.01)., Conclusions: 3D reconstruction technique could accurately simulate surgery procedure, calculate liver volume and evaluate the safety of hepatectomy for huge hepatocellular carcinoma. When the anatomical liver resection was performed, the larger tumor volume means the smaller excision functional liver volume and larger residual liver volume.

Objectives: To compare the outcomes after liver resection for a single small hepatocellular carcinoma (HCC) (≤ 5 cm) between non-cirrhotic patients and cirrhotic patients, and to explore the influence of liver cirrhosis on recurrence and overall survival after liver resection in patients with a single small HCC., Methods: A consecutive series of 256 patients with a single small HCC undergoing liver resection from April 2001 to October 2009 was retrospectively reviewed. Among the 256 patients, 227 patients were male, and 29 were female. The medium age was 49 years (ranged, 14 - 79 years); 224 (87.5%) patients were positive for hepatitis B surface antigen, 241 (94.1%) patients were with preoperative liver function of Child-Pugh grade A. The entire cohort were divided into non-cirrhosis group (n = 44) and cirrhosis group (n = 212). Univariate analysis and then multivariate analysis were performed to determine the prognostic factors of recurrence and overall survival after liver resection for all patients., Results: The 1-, 3-, 5-year recurrence-free survival rates after liver resection were 93.0%, 85.3%, and 68.5%, respectively, in non-cirrhosis group, while 81.1%, 58.6%, and 45.0%, respectively, in cirrhosis group. The 1-, 3-, 5-year overall survival rates after liver resection were 100%, 92.5%, and 92.5%, respectively, in non-cirrhosis group, while 93.8%, 78.7%, and 67.8%, respectively, in cirrhosis group. Both the recurrence-free survival and overall survival of non-cirrhosis group were significantly better than those of cirrhosis group (χ(2) = 8.756, P = 0.003; χ(2) = 8.603, P = 0.003). Cirrhosis, absence of tumor capsule, presence of microvascular invasion and moderate/poor tumor differentiation were the independent adverse prognostic factors for recurrence-free survival and overall survival in patients with a single small HCC after liver resection., Conclusions: Cirrhosis is an important adverse prognostic factor for long-term survival in patients with a single small HCC after liver resection. Liver resection resulted in much worse survival for cirrhotic patients compared to non-cirrhotic patients.

Objectives: To investigate the prognostic significance of Omi/HtrA2 expression, and the correlation between Omi/HtrA2 and Hypoxia-inducible factor (HIF)-1α in primary hepatocellular carcinoma cells., Methods: The expression of HIF-1α and Omi/HtrA2 in 43 cases of hepatic carcinoma tissues were detected immunohistochemically. Follow-up data were obtained to perform the Kaplan-Meier survival analysis. The change of Omi/HtrA2 expression in HepG2 cell was measured after HIF-1α expression of HepG2 in vitro was regulated by Tet-on expression system., Results: Omi/HtrA2 expression was correlated with lymph node metastasis and recurring within liver during 2 years. Statistical analysis estimation showed the cumulative survival rate of post-hepatectomy for the patients with the positive expression of Omi/HtrA2 was higher than that for other patients with the negative expression of Omi/HtrA2 (χ(2) = 6.13, P = 0.013). In the common paraffin-embedded specimen of hepatocellular carcinoma, most of the samples showing negative or weak positive HIF-1α immunopositivity showed moderate positive or strong positive Omi/HtrA2 immunopositivity, while most of the samples showing moderate positive or strong positive HIF-1α immunopositivity showed negative or weak positive Omi/HtrA2 immunopositivity. The mRNA expression intensity of Omi/HtrA2 was decreasing with the HIF-1α expression increasing, and the difference was statistically significant(F = 106.766, P < 0.01)., Conclusions: Omi/HtrA2 may be an important prognostic marker for primary hepatocellular carcinoma. Omi/HtrA2 expression is reversely correlated with HIF-1α expression in hepatocellular carcinoma. During the apoptotic process Omi/HtrA2 participating in hepatocellular carcinoma cells, HIF-1α is involved in the controlling and regulating of Omi/HtrA2 expression.

Objective: To determine whether hepatitis B virus X (HBX) protein expression affect the oval cells' response to anti-proliferative effect of transforming growth factor β1 (TGFβ1) in oval cells., Methods: Real-time PCR, Western blot analysis were performed to detect the expression of TGFβRII in HBX-transfected oval cells named HBX-EGFP-LE/6, and EGFP-LE/6, LE/6 control cells. In addition, exogenous TGFβ1 was added into all three oval cell lines, MTT assay was preformed to clarify different responses to the anti-proliferative effect of TGFβ1., Results: The TGFβRII mRNA levels in LE/6 and EGFP-LE/6 cells were (10.2 ± 1.8) and (8.8 ± 0.9) folds of those in HBX-EGFP-LE/6 cells, the difference was significant (P < 0.05). HBX protein expression also reduced the protein levels of TGFβRII in HBX-EGFP-LE/6 oval cells, compared to the control cells. The MTT results exhibited that, after TGFβ1 addition, proliferative inhibition rate in the HBX-EGFP-LE/6 cells was 18.1% ± 1.5% while those in control cells were 42.2% ± 2.8% and 41.9% ± 5.0%, the difference was significant (P < 0.01)., Conclusion: HBX protein expression affects TGFβRII transcriptional activity and protein synthesis, and insensitive oval cells to anti-proliferative effect of TGFβ1.

Objective: To investigate the changes of oval cell proliferation rate in the rat 2-acetylaminofluorene/partial hepatectomy (2-AAF/PH) model., Methods: Livers were collected from 2-AAF/PH rats at different time points after hepatectomy. Paraffin sections were investigated by double immunofluorescent staining with confocal microscopy for oval cell marker epithelial cell adhesion molecule and proliferative index proliferating cell nuclear antigen, or epithelial cell adhesion molecule and alpha-smooth muscle actin. Deposition of matrix in liver tissue was detected by sirius red staining., Results: Response of ductular oval cells could be observed in portal area at 2 days after PH, and the number of oval cells reached its peak at 9 days and then gradually declined. Oval cell proliferation rate decreased from (91.3 +/- 1.6)% at 2 days after PH to (53.6 +/- 4.4)% at 12 days (P < 0.01). In addition, oval cells infiltrating into liver parenchyma were closely associated with activated hepatic stellate cells and extracellular matrix., Conclusions: Oval cell proliferation rate starts decreasing before its number reaches a peak in 2-AAF/PH model. Hepatic stellate cells probably tightly regulate oval cell number through secreting several factors and producing extracellular matrix.

Objective: To find out the mechanisms of HBx gene inducing oval cell malignant transformation into hepatoma carcinoma cell., Methods: The changes of morphology, cell cycle, differentiated markers, c-myc and TGF-alpha in pEGFP-HBx oval cells strain, which stably expressed HBx gene, were studied by inversion phase contrast microscope and transmission electron microscopy, flow cytometry, periodic acid-schiff (PAS) staining, soft agar growth assay, real-time PCR, immunocytochemistry. pEGFP-oval cells and LE/6 oval cells were used as control groups., Results: (1) The pEGFP-HBx oval cells showed bigger in size with malformed nucleus as compared with control groups. (2) Flow cytometry showed that, in contrast with the control groups, the proportion of pEGFP-HBx oval cells arrested in G0/G1 phase decreased but in S or G2/M phase rose. Moreover, the population of aneuploid cells increased obviously. (3) PAS staining showed that there were many glycogen granules in the cytoplasm of pEGFP-HBx oval cell. (4) The pEGFP-HBx oval cell formed colonies in the soft agar. (5) Compared with the control groups, the expression of HNF-4 alpha, AFP, c-myc and TGF-alpha rose obviously, whereas the expression of CK-7 and CK-19 decreased. And the expression of cps1 mRNA was not in the extent of detection., Conclusions: The HBx gene can provoke abnormal differentiation of oval cell and induce oval cell malignant transformation.

Objective: To evaluate long-term outcomes of minor liver resection for hilar cholangiocarcinoma (HC) of Bismuth-Corlette type III., Methods: From January 1997 to December 2007, the clinical data of 91 patients with Bismuth-Corlette type III HC underwent hepatectomy were collected and analyzed retrospectively., Results: There were 60 patients underwent minor hepatectomy, and 31 undergoing major hepatectomy. Hepaticojejunostomy was made conventionally in an end-to-side fashion in the patients undergoing major liver resection, and a new technique of hepaticojejunostomy used in the patients undergoing minor liver resection. That was the anterior edges of bile duct stumps which were not sutured after suturing of posterior edges. Instead of, the anterior edge of jejunum loop to the remnant liver on the top of the bile duct stumps were sutured with intermittent "U" sutures. In all patients, in-hospital mortality rate was 0 and rate of bile leakage was only 2.1%. The actual 1-, 3- and 5-year survival rates were 91.6% and 87.0%, 61.6% and 62.0%, 31.6% and 33.0%, respectively (P > 0.05)., Conclusions: Minor liver resection for the selected patients with HC of Bismuth-Corlette type III according to our criteria achieved better long-term outcomes. A new hepaticojejunostomy used in the patients undergoing minor liver resection is a safe and effective method.

Objective: To study the effect of Extractum trametes robiniophila murr on cardiac allograft rejection in mice., Methods: All abdominal heterotopic heart transplantation models were divided into three groups as follows: (A) Extractum trametes robiniophila murr group. (B) Rejection group. (C) Isograft group. In each group, mean survival times (MST) of transplanted hearts and their pathologic histological changes at postoperative fifth day were observed. With fluoroimmunoassay, granzyme B and CD8(+) expressions were examined., Results: The MST of heart allografts in group A were (6.38 +/- 0.69) d, significantly shorter than that of group B [(8.31 +/- 0.59) d] (P < 0.01). In group A, acute rejection was present in advance; transplanted hearts were seriously damaged into acute rejection pathological grade 3, and CD8(+) T lymphocytes infiltrated diffusely and the expression of granzyme B increased significantly as compared with other groups., Conclusions: Exclusive application of Extractum trametes robiniophila murr can promote the acute rejection of graft in early phase of postoperation, and the mechanism may be the promoted proliferation and infiltration of CD8(+) T lymphocytes and the increased expression of granzyme B.

Objective: To retrospectively analyze the experiences, indications, technique, and results of coronary artery bypass grafting (CABG) in patients over 70 years old., Methods: Ninety-one patients received coronary artery bypass grafting from March 2004 to March 2008. Ages ranged from 70 to 83 years old, 22 patients over 75 years old. Conventional CABG (CCABG) in 72 patients, off-pump CABG (OPCAB) in 19 patients. Clinical data has no significant differences in two groups. The rate of using left internal mammary artery was 96.7%. The number of grafts in CCABG and OPCAB group were 2 to 5 (3.5 +/- 0.8) and 1 to 4 (2.9 +/- 0.7) respectively., Results: In-hospital death in 2 cases, both were from chronic obstructive pulmonary disease and pulmonary infection. Cerebral infarction in 1 case and pulmonary infection in 2 cases in CCABG group, but no significant difference between two groups, and no difference in intubation, ICU stay, respiratory failure, renal function failure. But number of grafts in CCABG was significantly more than that in OPCAB (P < 0.01). Postoperative follow-up was 3 to 36 months, 1 case with recurrent angina in OPCAB., Conclusions: According to the characteristic of coronary artery disease in elderly, fully revascularization and improving myocardial blood supply, patients over 70 years old with CABG can obtain the same efficacy as younger patients. There were not significant difference between CCABG and OPCAB.

Objective: To review the experience of staged total cavopulmonary connection (TCPC) in complex congenital heart diseases., Methods: From June 1998 to March 2008, 22 patients underwent staged TCPC for complex congenital heart diseases. Among them, 9 were univentricular and pulmonary artery valve stenosis; 3 were univentricular and pulmonary artery atresia; 1 was transposition of great arteries, crisscross heart and pulmonary artery valve stenosis; 1 was complete atrioventricular canal defects, left ventricular hypoplasia, pulmonary artery atresia and atrioventricular valvular regurgitation; 1 was complete atrioventricular canal defects, left ventricular hypoplasia, pulmonary artery valve stenosis and atrioventricular valvular regurgitation after Glenn procedure; 1 was mirror image dextrocardia, single ventricle, pulmonary artery atresia, major aortopulmonary collateral arteries (MAPCAs) and right pulmonary arteriovenous fistula after Glenn procedure; 4 were tricuspid atresia and pulmonary artery valve stenosis; 1 was tricuspid atresia and pulmonary atresia; 1 was mirror image dextrocardia, double-outlet of right ventricle, left ventricular hypoplasia, pulmonary artery valve stenosis, tricuspid incompetence, and MAPCAs. Among them, 5 patients received systemic-to-pulmonary artery shunt, bidirectional Glenn procedure and TCPC. Seventeen patients received bidirectional Glenn procedure, the mean age was (5.9+/-4.4) years old. Pulmonary artery pressure pre-Glenn procedure was 17 to 20 mm Hg (1 mm Hg=0.133 kPa). Atrioventricular valve incompetence in 3 patients. Nakata index was less than 200 mm2/m2 in 4 patients before the first stage operation. The age of TCPC procedure was (9.6+/-4.9) years old, the interval time was (3.7+/-1.2) years., Results: There was one in-hospital death, the mortality was 4.5%. The patient with univentricular and pulmonary atresia, received systemic-to-pulmonary artery shunt, bidirectional Glenn procedure and TCPC and died of pneumorrhagia. Other patients were recovered well, postoperative central venous pressure was 12 to 18 mm Hg, percutaneous oxygen saturation was 90% to 96%. The cardiac function were in NYHA class I to II., Conclusions: The staged TCPC was a good procedure in high-risk Fontan candidates. The results were satisfactory for those patients. This staged strategy may extend the operative indications for the Fontan procedure.

Objective: To construct pEGFP-HBx eukaryotic expression plasmid and establish stable and effective transfected rat oval cell (LE/6) strain expressing EGFP-HBx fusion protein to explore the roles of HBx gene and oval cell in carcinogenesis of hepatocellular carcinoma (HCC)., Methods: HBx gene with EcoRI and Hind III endo-enzyme sites was obtained by using PCR from plasmid pcDNA3.1-HBx. The purified HBx gene fragment was inserted into pEGFP-N1 expression vector, and the recombinant plasmid pEGFP-HBx was identified by restriction endonuclease and DNA sequencing analysis. LE/6 cells were transfected with recombinant pEGFP-HBx by lipofectamine reagent. Resistant to G418 clones were selected, and expression of EGFP-HBx fusion protein in clones were examined directly with fluorescence microscope, and these clones were isolated and proliferated. The expression of HBx was detected by RT-PCR analysis and immunocytochemistry., Results: Plasmid pEGFP-HBx has whole HBx gene base and correct reading frame as indicated by restriction endonuclease and DNA sequencing analysis. After transfecting with pEGFP-HBx plasmid, LE/6 cell clones expressing EGFP-HBx fusion protein were obtained. RT-PCR analysis and immunocytochemistry showed that HBx gene was only expression in transfected pEGFP-HBx cells., Conclusions: The pEGFP-HBx recombinant expression vector was successfully constructed, and the stable transfected LE/6 strain expressing EGFP-HBx fusion protein was successfully established. It will be helpful in the further study on the roles of HBx and liver oval cell in carcinogenesis of HCC.

Objective: To study the function of nuclear factor-kappaB (NF-kappaB) signaling pathway in regulating vascular endothelial growth factor (VEGF) by hepatitis B virus X protein (HBx)., Methods: After the establishment of L02-HBx cell line with stable transfected HBx gene, NF-kappaB signaling pathway blocker PDTC was introduced to cut off its signal transduction. Double immunofluorescent staining and laser scanning confocal microscopy were applied to study the activation and deactivation of NF-kappaB signaling pathway. Real-time PCR and Western blot were used to observe the expression of VEGF gene before and after the HBx transfection, as well as the treatment with PDTC., Results: The NF-kappaB signaling pathway of L02-HBx cells was activated after transfection with HBx gene as compared to the control L02 cells without treatment. The mRNA and protein levels of VEGF in L02-HBx cells increased 4.07 +/- 0.31 and 4.34 +/- 0.64 times respectively. The difference was of statistical significance (P < 0.05) in comparison with the control cells. The mRNA levels of VEGF decreased to 2.33 +/- 0.22 and 1.86 +/- 0.18(P < 0.05) and at the same time the expression of VEGF also reduced to 2.52 +/- 0.29 and 2.17 +/- 0.34 (P < 0.05), after treatment with 25.0 micromol/L and 50.0 micromol/L PDTC for 24 h respectively when the NF-kappaB signaling pathway was blocked. There was no significant difference in VEGF mRNA and protein levels when treated with 12.5 micromol/L PDTC for 24 h., Conclusion: NF-kappaB signaling pathway maybe one of the routes through which HBx up-regulate the expression of VEGF to promote angiogenesis in hepatocellular carcinoma.

Objective: To study location of MT1-MMP and effect of its change in expression on rabbit VX2 tumor tissues after transarterial embolization with hydroxyapatite nanoparticles loaded with lipiodol., Methods: Sixty rabbits implanted with tumor tissue of cell line VX2 were divided into three groups (control group, lipiodol group, hydroxyapatite nanoparticles loaded with lipiodol group). The transarterial embolization was performed super-selectively via gastro- duodenal artery of rabbits, each rabbit in control group was inserted with 1 ml normal saline,that in lipiodol group was inserted with 0.3 lipiodol ml/kg, also 0.3 ml hydroxyapatite nanoparticles loaded with lipiodol per kg for that in the last group. Results of embolization were detected by using CT scanning 3 days after operation. After two weeks, all tumors were took out as specimens to investigate location of MT1-MMP in VX2 tumor tissues,and also to determine the change of its expression in tumor tissues after embolization with different medicines, with three-step immunohistochemical technique (S-P). MT1-MMP mRNA was measured by RT-PCR to determine whether there were differences in three groups. Western blot technique was performed to determine difference of MT1-MMP protein expression of in three groups., Results: Immunohistochemical results exposed that MT1-MMP was expressed on membrane of tumor cells and in extracellular matrix of tumor cells. Comparison of MT1-MMP expression in control group with that in other two groups, showed a significant lower level in control group (P < 0.05). There was no difference in MT1-MMP expression between lipiodol group, hydroxyapatite nanoparticles loaded with lipiodol group (P > 0.05). Western blot supported this conclusion. RT-PCR detecting MT1-MMP mRNA was found no differences among three groups (P > 0.05)., Conclusions: MT1-MMP was mainly expressed on membrane of tumor cells and in extracellular matrix of tumor cells. There was an increasing tendency on expression of MT1-MMP in tumor tissues and extracellular matrix after transarterial embolization with hydroxyapatite nanoparticles loaded with lipiodol,it might be one of important mechanisms provoking high recurrence rate for hepatocellular carcinoma after treatment embolization.

Objective: To study the inducible expression of hypoxia-inducible factor 1alpha (HIF-1alpha) on the proliferation and invasion property of HepG2 cells under normoxia in vitro., Methods: Constructed the HepG2(Tet-on-HIF-1alpha) cell line which could induce the expression of HIF-1alpha by doxycycline; Under normoxia in vitro, MTT assay was used to observe the proliferative and adhesive activity of cells, and the invasive activity was determined by transwell cell culture chamber method., Results: Under normoxia, the HIF-1alpha mRNA and protein of HepG2(Tet-on-HIF-1alpha) cells could be induced up to (5.899 +/- 2.176) and (2.179 +/- 0.742) folds by doxycycline (1 microg/ml); There were no difference of A(490 nm) between the Dox(+)and Dox(-) group in experiment detecting the proliferation activity (P > 0.05); But in adhesive experiment, the A(490 nm) of Dox (+) group was 0.662 +/- 0.058, higher than the Dox(-) group 0.526 +/- 0.808 (P = 0.008); The invasive cell number of Dox(+) group was 37.611 +/- 8.424, but in the Dox(-) group, the number was 25.333 +/- 8.117 (P < 0.01)., Conclusions: Under normoxia in vitro, the Tet-on gene regulate system could increase the HIF-1alpha protein by inducing the HIF-1alpha mRNA; HIF-1alpha has no influence with the proliferation activity, but it could enhance the adhesive and invasive properties of HepG2 cells.

Objective: To elucidate intracellular signal pathway in formation of multidrug resistance (MDR) of hepatocellular carcinoma (HCC) induced by its microenvironment, and to explore the potential role of extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) pathway in this process., Methods: Activity of ERK/MAPK was examined by Western blot technique through comparing the ratio of phosphorylation of ERK/MAPK to total ERK/MAPK protein in HepG2 cells exposed to hypoxia, low glucose or transfected by plasmid pcDNA3/HBX. After being treated by the specific ERK/MAPK pathway inhibitor U0126, Western blot technique was used to analyze the alterations of the expression of P-gp, MRP1, LRP and HIF-1alpha at protein level. RT-PCR was used to analyze the alterations of the expression of HIF-1alpha mRNA. Cellular location of HIF-1alpha protein was determined by immunocytochemistry after being treated by U0126., Results: The activations of ERK/MAPK determined by the ratio of phosphorylated ERK/MAPK to the total ERK/MAPK were increased in varying degrees in HepG2 cells respectively exposed to different microenvironment. After being treated by U0126 for 12 h, the expressions of mdr1, MRP1, LRP genes and protein in those cells were decreased to some extent. However, the gene expression of HIF-1alpha was not influenced and only its protein was decreased. HIF-1alpha protein was reversely translocated into cytoplasm from nucleus after being treated by U0126., Conclusions: ERK/MAPK pathway is involved in the course of the formation of MDR of HCC induced by microenvironment.

Objective: To report the experience of extracardiac conduit total cavopulmonary connection (ECTCPC) in surgical treatment of complex congenital heart diseases., Methods: From 1998 to 2006, 68 patients underwent ECTCPC for complex congenital heart diseases. Among them, 45 had functional univentricle with transposition of the great artery (TGA) and pulmonary artery valve stenosis, 19 had tricuspid atresia with hypoplasia of right ventricle, 4 had Ebstein's anomaly with hypoplasia of right ventricle. Six had left superior vena cava, 18 had received Bidirectional Glenn operation; Fifty-seven cases were performed under cardiopulmonary bypass with general anesthesia and hypothermia, 11 cases were performed without cardiopulmonary bypass., Results: There were two death, the mortality was 2.9%. All patients were followed up from 1 to 8 years with no clinical symptoms and have been doing well. The arterial oxygen saturation was 90% - 96%, the cardiac function were in NYHA class I - II., Conclusion: The extra cardiac conduit TCPC is a simple procedure and superior to other type of Fontan procedure in most patients.

Objective: To investigate the reversal of multidrug resistance in the cell line HepG2/ADM induced by TNF-alpha., Methods: HepG2/ADM cells were incubated with different concentrations of TNF-alpha (100, 500 and 2500 U/ml) for 72 h. Real-time PCR was performed to compare the mRNA levels of MDR1 with PPAR-alpha in the different concentrations of TNF-alpha treated cells. The Annexin V assay was used to check cell apoptosis induced by 0.5 mg/L adriamycin. Rhodamine 123 efflux assay and MTT assay were used to study P-gp activity and drug resistance in each group, respectively., Results: TNF-alpha could induce down-regulation of MDR1 and up-regulation of PPAR-alpha. Meanwhile, it could enhance cell cytotoxicity and cell apoptosis induced by 0.5 mg/L adriamycin., Conclusions: TNF-alpha could partially reverse the multidrug resistance of HepG2/ADM cells by down-regulating the expression of MDR1 and up-regulating the expression of PPAR-alpha.

Objective: To sum up the experience of diagnosis and treatment of rare hepatic benign and malignant occupying lesion (OL) in 42 cases., Methods: The clinical data of 42 cases of rare hepatic benign and malignant OL were reviewed retrospectively., Results: Accordance rate of the imaging diagnosis with pathological result was 23.8%; the diagnostic accuracies of ultrasound B, CT and MRI were 14.3%, 19.4% and 22.2%, respectively. All cases were followed up for 1 - 5 years, no recurrence was found in 32 cases of benign OL post operation, 2 patients with malignant OL has survived for 18 and 36 months respectively, other 5 survived an average time of 7.5 months., Conclusions: It's difficult to make a confirmed diagnosis of rare hepatic benign and malignant OL before operation, combined examination of ultrasound B, CT and MRI can improve accuracy in diagnosis, no recurrence occurred in benign OL after operation, surgery can prolong survival in malignant OL.

Objective: To investigate the change of chemosensitivity of hepatocarcinoma cell line (HepG(2)/ADM) after treated by bromocriptine (BCT) combination with human tumor necrosis factor-alpha (TNF-alpha)., Methods: Firstly, TNF-alpha gene was transfected into HepG(2)/ADM cell line by liposome to establish a cell model expressing the TNF-alpha protein stably. All experiments were divided into four groups and named blank control group (group A), drug resistant group HepG(2)/ADM (group B), TNF-alpha gene group HepG(2)/ADM/TNF (group C) and BCT group (group D) respectively. And group D came from group C treated with BCT simultaneously. MTT assay was tested to detect the sensitivity to ADM of each group and Rhodamine 123 (Rh123) was applied to test the function of P-gp by flow cytometric analysis (FCM). MDR associated genes and proteins and PKC-alpha protein were detected by immunohistochemistry (IHC), Western blot and reverse transcriptase polymerase chain reaction (RT-PCR) methods, respectively. The expression and the apoptosis rate of Bcl-2 in the hepatocarcinoma cells were detected by FCM., Results: There was significant difference between group C and D in the rate of reversing resistance and the intracellular Rho123 accumulation (P < 0.01). MDR1 mRNA and P-gp protein expression in group C and D were low similar to that in group A, but no difference could be found among them (P > 0.05). As we found that PKC-alpha protein expression was downregulated in group D but Bcl-2 protein expression was downregulated in group C, and there was significant difference compared to other groups. The apoptosis rate of hepatocarcinoma cells was much higher in group D than that in group C (P < 0.01) with FCM, but similar to group A (P > 0.05)., Conclusions: Synergistic effect of BCT and TNF-alpha on reversing hepatocellular carcinoma multidrug resistance could enhance the susceptibility of HepG(2)/ADM cells to cytotoxic drugs.

Objective: To investigate the expression and clinicopathological significance of transcription factor Twist in hepatocellular carcinoma (HCC), paraneoplastic and cirrhotic tissues., Method: Immunohistochemistry was used to detect the expression of the Twist protein in 26 cases of HCC and paraneoplastic tissue and 10 cases of cirrhotic tissue. Meanwhile, the Twist mRNA and its protein were detected in 10 HCC tissues and 10 paraneoplastic tissues by RT-PCR and Western blot. And the expression differences and clinicopathological significances of the expression of Twist gene and its protein were analyzed., Results: The positive rates of the Twist protein in HCC, paraneoplastic and cirrhotic tissues were 84.6%, 19.2 % and 20.0 %, respectively. The positive rate of Twist in HCC was higher than that in paraneoplastic or cirrhosis tissues (P < 0.05). Compared with in paraneoplastic tissues, the mRNA and protein expression of Twist were up-regulated in HCC by 2.52 and 2.13 times, respectively., Conclusion: Twist has an inappropriate expression in hepatocellular carcinoma and it may play an important role in the tumorigenesis and progression of HCC.

Objective: To investigate the effects of tumor necrosis factor related apoptosis-inducing ligand (TRAIL) combined with Chinese medicine, Extractum trametes robiniophila murr in human hepatic cancer cells., Methods: HepG2 cell line resistant to adriamycin (HepG2/ADR) was induced step by step. The effects of TRAIL (100 ng/L) combined with Extractum trametes robiniophila murr (1.0 g/L) promoting apoptosis in HepG2 or HepG2/ADM were analyzed. The proliferation was observed by MTT assay and the apoptosis of cells was also observed by flow cytometry., Results: HepG2/ADM was confirmed resisting to ADM. The treatment of TRAIL (100 ng/L) combined with Extractum trametes robiniophila murr (1.0 g/L) showed significant inhibitory effects on the growth of both HepG2 and HepG2/ADM, and the percentage of apoptosis was increased compared with other groups within 24 to 72 h., Conclusions: Extractum trametes robiniophila murr dramatically augmented the sensitivity of both HepG2 and HepG2/ADM to TRAIL, but only has slightly killing effects on L02. TRAIL combined with Chinese medicine treatment could be a safe and attractive strategy to drug-resistant/TRAIL-resistant tumor cells.

Objective: To investigate synergistic effect of bromocriptine (BCT) combining tumor necrosis factor-alpha (TNF-alpha) on reversing multidrug resistance in a nude mouse model of liver neoplasm., Methods: Human hepatocarcinoma cell line HepG(2) (HepG(2) group), drug resistant hepatocarcinoma cell line HepG(2)/adriamycin (HepG(2)/ADM group) and hepatocarcinoma cell line transfected with TNF-alpha gene HepG(2)/ADM/TNF (TNF group and BCT group) were injected into the liver of nude mice via orthotopic implantation to establish multidrug resistance model of liver neoplasm in vivo. All the mice were injected with 5-fluouracil + adriamycin + mitomycin in abdominal cavity for 7 d. The mice in BCT group was simultaneously given bromocriptine through gastric canal. Size and weight of the tumor were measured. Furthermore tumor histological character and growth of the nude mice was observed and its chemosensitivity was tested. MDR associated genes and proteins (MRP, LRP) of implanted tumors were detected by immunohistochemical staining and reverse transcriptive polymerase chain reaction (RT-PCR), and apoptosis rate of hepatocarcinoma cells was detected by TUNEL assay., Results: The nude mouse model of each cell line was all inoculated successfully. The tumor growth rate and weight were significantly different among groups (P < 0.05). After chemotherapy tumor growth inhibition rate was higher in BCT group (67%) compared to ADM and TNF groups (P < 0.01), and similar to HepG(2) group (54%). MDR1 and LRP mRNA could be detected in all groups, but TNF-alpha was detected only in TNF-alpha and BCT groups. Furthermore, MDR1 and LRP protein expression of tumors in TNF-alpha and BCT groups was low similar to HepG(2) group. The apoptosis rate of hepatocarcinoma cells was much higher in BCT group than in other groups (P < 0.05) with TUNEL assay., Conclusions: TNF-alpha gene can down-regulate the MDR associated genes and proteins expression for example MDR1, LRP, and lower its tumorgenesis. Moreover, bromocriptine can enhance the susceptibility of HepG(2)/ADM cells to cytotoxic drugs.

Objective: To investigate the role of united hepatectomy and splenectomy in the surgical treatment of hepatocellular carcinoma complicated with hepatic cirrhosis and hypersplenism., Methods: Two hundred and four patients of hepatocellular carcinoma complicated with liver cirrhosis and hypersplenism were divided into two groups: the group of combined resection of hepatocellular carcinoma and spleen (group A, n = 94) and the group of hepatectomy only (group B, n = 110). The counts of white blood cell and platelet, total serum bilirubin levels, changes of immune function, operative morbidity and 5-year survival rates were compared between the two groups., Results: (1) There was no significant difference of the counts of CD4, CD8, CD4/CD8 and the levels of IL-2, IFN-gamma and IL-10 between the two groups before the operation. (2) Two months after operation, the percentage of CD4 and the ratio of CD4/CD8 were significantly higher in the group A [(40.8 +/- 4.1)% and (1.8 +/- 0.2)%, respectively] than those of group B [(33.8 +/- 3.6)% and (1.1 +/- 0.3)%, respectively], while the percentage of CD8 was (25.8 +/- 3.8)% in the group A, significantly lower than that of group B [(32.9 +/- 4.1)%, P < 0.05]; Both the levels of IFN-gamma and IL-2 were significantly higher in the group A than those of group B while the level of IL-10 in group A was lower compared with that of group B (P < 0.05). (3) On the 14 postoperative day, the counts of white blood cell and platelet were (9.1 +/- 1.4) x 10(9)/L and (310 +/- 55) x 10(9)/L, which were significantly higher than those of group B [(3.6 +/- 1.2) x 10(9)/L and (99 +/- 36) x 10(9)/L, respectively]. (4) On the 7th postoperative day, the total serum bilirubin concentration of group A [(24 +/- 7) micromol/L] was lower than that of group B [(37 +/- 13) micromol/L]. (5) There was no significant difference in the postoperative morbidities between the two groups (15.9% and 14.5%, respectively). (6) There was no significant difference of the 5-year cumulative survival rates between group A (56.4%) and group B (50.9%, P > 0.05), but the survival rate without tumor of group A was 37.7%, higher than that of group B (18.9%, P < 0.05)., Conclusions: The combined resection of hepatocellular carcinoma and spleen for the hepatocellular carcinoma complicated with liver cirrhosis and portal hypertension may promote the recovery of the balance between the subgroup of T cell and B cell, normalize the counts of white blood cell and platelet, alleviate the bilirubin burden and benefit for the recovery of liver physiological role without increase; the 5-year disease-free survival rate was improved significantly while no increase of postoperative morbidity. Combined resection may also be helpful for the delay of the progression of liver cirrhosis and for the prevention of esophageal variceal bleeding.

Objective: To explore the mechanism of multidrug resistance of hepatocellular carcinoma induced by hypoxia and the potential role of hypoxia-inducible factor-1 alpha (HIF-1 alpha) and multidrug resistance related genes., Methods: Human hepatocarcinoma cell lines HepG2 cells were exposed to hypoxia and were transfected by plasmid HIF-1 alpha/PCDNA3, respectively. The expressions of multidrug resistance gene (mdr1), multidrug resistance protein (MRP1), and lung resistance protein (LRP) gene at the mRNA and the protein levels in the above two groups were respectively analyzed by real-time fluorescent quantitative PCR and Western-blot technique., Results: In the hypoxia group, the expressions of mdr1, MRP1 and LRP were stepped up correlating to the degree of hypoxia, especially the prominent increase in the expression of MRP1. Furthermore, they were synchronous with the changes of the expression of HIF-1 alpha. Also the increased expression of mdr1, MRP1, and LRP gene was observed in transfected HepG2 cells by plasmid HIF-1 alpha/PCDNA3., Conclusions: Resistance of hepatocellular carcinoma to chemotherapeutics could be induced by hypoxia. HIF-1 alpha may be critical to the upregulation of the expression of the related multidrug resistance genes induced by hypoxia. HIF-1 alpha and these related multidrug resistance genes could be potential molecular targets for reversing multidrug resistance of hepatocellular carcinoma.

Objective: To explore the relationship between kruppel-like factor (KLF)6 gene and the development or progression of hepatocellular carcinoma (HCC)., Methods: Reverse-transcription polymerase chain reaction (RT-PCR) was used to examine the expression of KLF6 mRNA in normal liver tissue and primary hepatocellular carcinoma, and single strand conformation polymorphism (SSCP), DNA sequencing were used to detect the point mutation of KLF6 in primary hepatocellular carcinoma., Results: An amplified fragment of 427 bp DNA was detected in 31 (97%) of 32 adjacent noncancerous tissue and normal liver tissue, and in 23 (85%) of 27 HCCs. There was no significant difference in the levels of KLF6 mRNA between normal liver and liver tumors (chi(2) = 2.58, P > 0.05). For the 27 HCCs, six SSCP-positive bands (22%) were detected. Among them, three of 5 (3/5) tumor samples showing loss of heterozygosity (LOH) of KLF6 had mutations in the retained KLF6 allele., Conclusion: We showed that LOH was detected in 5 (36%) HCCs obtained from 14 informative cases, and three of 5 tumor samples showing LOH of KLF6 had mutations in the retained KLF6 allele. Two inactivating events had occurred; thus, as defined by Knudson's "two-hit model", 16 KLF6 appears to be a tumor suppressor gene.

Objective: To reverse multidrug resistance (MDR) of HepG2 by anti-MDR1 hammerhead ribozyme., Methods: We developed an anti-MDR1 hammerhead ribozyme and delivered it to P-gp-overproducing human hepatocarcinoma cell line HepG2 by a retroviral vector containing RNA polymerase III promoter. We detected the expression of MDR1/Pgp and Rz in HepG2, HepG2 multidrug-resistant cell line and HepG2 Rz-tranduced cells by real-time RT-PCR, semi-quantitative RT-PCR and western blot methods. Moreover, MTT assay was tested to detect sensitivity of these ribozyme-tranduced cells, and Rhodamine123 (Rh123) applied to test the function of Pgp., Results: The Rz-tranduced HepG2 cells became doxorubicin-sensitive, concomitant with the decreases in MDR1 expression, P-gp amounts and efflux pump function., Conclusions: The approaches using either retrovirus or liposome-mediated transfer of anti-MDR1 ribozyme may be selectively applicable to the treatment of MDR cells.

Objective: To investigate the effects of HBx gene on proliferation activity of hepatoma cells in vitro and in vivo., Methods: The plasmid pHA-HBx carrying HBx gene was transfected into HepG(2) cells, and the positive clones were screened and identified with G418 and RT-PCR, respectively. The growth curve and population doubling time were calculated, and the cell cycle was analyzed by flow cytometry (FCM). The proliferation activity of transformed cells was measured with (3)H-TdR incorporation rate and nude mice model in vitro and in vivo., Results: The result of RT-PCR indicated that HBx gene was integrated into the genome DNA of HepG(2) cells and transcripted. The growth curve and population doubling time showed a high proliferation activity of transformed cells. The amount of cells at stage S and G(2)/M were significantly higher, and cells at stage G(0)/G(1) were lower than those in control group. The tumors developed from transfected cells grew much quicker than those developed from HepG(2) cells in nude mice model., Conclusion: HBx gene can facilitate the proliferation of hepatoma cells both in vitro and in vivo.

Objective: To study the therapeutic potential of TRAIL and in combination with subtoxic level of chemotherapeutic agents in the treatment of human hepatocellular carcinomas (HCCs)., Methods: The plasmid pcDNA 3.0-hTRAIL was transfected into COS-7 cells, and it was transiently expressed. The cytotoxic functions of the expressed product and in combination with chemotherapeutic agents were detected., Results: The transiently transfected COS-7 could express the active human TRAIL. It had mild cytotoxic functions on HCC cell lines. The cytotoxicity rates in both cell lines were 9.2% and 9.5% respectively. Chemotherapeutic agents, mitomycin (M), 5-FU and actinomycin D (AcD) dramatically augmented TRAIL induced cytotoxic function. There were significant differences in the cytotoxicity between the use of single agent and in combination with TRAIL (TRAIL + M, M; 60.1%, 32.6%, TRAIL + 5-FU, 5-FU: 68.1%, 29.2%, TRAIL + AcD, AcD; 72.9%, 58.6%) (P < 0.05)., Conclusions: The combination of human TRAIL and chemotherapeutic agents, such as mitomycin, 5-FU and actinomycin D, seemed to exert a synergistic effect compared with either agent alone, and it might have therapeutic potential in the treatment of human HCC.

Objective: To set up a mdr1 multidrug resistant model of orthotopic transplantation of liver carcinoma in nude mice by intermittent abdominal chemotherapy., Methods: The hepatocellular carcinoma cell line HepG2 was first cultured, then subcutaneous carcinoma was produced to form the tumor donor mice. An orthotopic mdr1 hepatoma was produced by implanting the tumor fragment subserosally to the mice liver, and intermittent chemotherapy with Pharmorubicin given to induce drug resistance. Physical examination, ultrasonography, spiral CT and laparotomy were used to examine the growth of the tumor. RT-PCR and SP method by the monoclonal antibody JSB-1 were adoped to detect the expression of mdr1-mRNA and p-gp protein., Results: There was no mortality. The successful rate of tumor implantation is 88% (22/25), the successful rate of supplementary implantation was 100% (3/3), and the successful rate of induction of drug resistance was 80% (16/20). The expressions of mdr1-mRNA and p-gp in the Pharmorubicin group were 23 folds and 13 folds of the control group, respectively., Conclusions: The nude mice mdr1 multidrug resistant model of orthotopic liver carcinoma were set up successfully, with its features similar to clinical cases. It would be helpful for the further study of the diagnosis and reversal strategy of the MDR phenomenon.

Objective: To investigate therapeutic potential of TRAIL in hepatocellular carcinoma (HCC) and the mechanism of sTRAIL resistance and to reverse the resistance to sTRAIL-inducing apoptosis., Methods: The expression profiles of TRAILR were determined 60 HCC samples, in 20 normal liver tissues and 2 HCC cell lines HepG2 and SMMC-7721 by in situ hybridization. Cellular effects of sTRAIL in promoting apoptosis on HCC cell lines HepG2 and SMMC-7721 were analyzed after exposure to recombinant protein and after transfection with a cDNA expression construct. In vivo effects of sTRAIL on tumor growth were investigated using a nude mice HCC model of hepG2. Furthermore, the expression of survivin in HCC was detected, and treatment with antisence oligonucleotide was accepted. Finally, therapeutic effect on HCC by combining sTRAIL and interleukin-12 (IL-12) was detected., Results: Both DR4 and DR5 were present in all HCC tissues as well as normal hepatic tissues. In contrast, 54 HCC tissues did not express DcR1 and 25 did not express DcR2. But both DcR were detectable in all of the normal liver tissues. The expression patterns of DR and DcR in HCC samples were quite different from those in normal tissue. DR5, DR4, and DcR2 expressed in both cell lines, while no DcR1 expression was detected. Recombinant sTRAIL alone was found to have a slight activity as it killed a maximum of 15% of HCC cells within 24 h while killing over 70% of Jurkat cells. In vivo administration of the TRAIL gene couldn't inhibit tumor growth in a nude mice HCC model. Mostly, HCC tissue and both HCC cell lines expressed survivin, whereas normal liver tissue did not express survivin. Treatment with antisence oligonucleotide enhanced sTRAIL-inducing apoptosis. IL-12 significantly augmented sTRAIL-inducing apoptosis and inhibited survivin expression., Conclusions: HCC cells are insensitive towards TRAIL-mediated apoptosis. Survivin may play a role in resistance to TRAIL-induced apoptosis in HCC, and antisence oligonucleotide could partly reverse the resistance to TRAIL-inducing apoptosis. IL-12 may sensitize HCC cells to TRAIL-induced apoptosis by preventing survivin. Combining gene therapy strategy such as combining gene therapy of TRAIL with IL-12 may be a promising maneuver to HCC.

Objective: To evaluate wound healing after types of pancreaticojejunostomy., Methods: After resection of the pancreatic head, 38 domestic piglets were divided into two groups according to the types of anastomoses: group I: binding pancreaticojejunostomy, a new technique designed and advocated by professor Peng Shuyou; group II: end-to-end pancreaticojejunal invagination. Anastomotic strength in vivo and histopathological findings were assessed on operative day and postoperative day 5 and 10., Results: Bursting pressure was 139.7 +/- 8.0, 178.7 +/- 9.7 and 268.8 +/- 12.8 mm Hg in group I on day 0, 5 and 10, whereas 67.3 +/- 7.9, 96.2 +/- 10.4 and 130.6 +/- 9.3 mm Hg in group II. The gain on day 0 to 5 and 5 to 10 was 27.9% and 50.5% in group I and 42.9% and 35.7% in group II, respectively. A significant difference was observed between group I and group II, and between 5 and 10 day after anastomoses (P < 0.01). Breaking strength was 4.5 +/- 0.4, 6.6 +/- 0.4 and 10.0 +/- 0.6 N in group I on day 0, 5 and 10 and 4.6 +/- 0.6, 5.8 +/- 0.5 and 7.1 +/- 0.6 N in group II. Although a similar value was shown in both types of anastomoses on day 0, a rapider gain was demonstrated on day 0 to 5 and 5 to 10 in group I (44.8% and 52.9%) than in group II (25.4% and 22.0%). A significant difference was found on day 5 and 10 between the two types of anastomoses (P < 0.05 and P < 0.01). Anastomotic site was well repaired by connective tissue and the cut surface of pancreatic stump was covered by mucosal epithelium in group I on day 10, but the cut surface was incompletely repaired by granulation tissue and no, regeneration of the epithelium was found in group II., Conclusion: Anastomotic strength of binding pancreaticojejunostomy was stronger than end-to-end pancreaticojejunal invagination and the healing was better and rapid.

Objective: To analyse the causes and the management of massive hemorrhage in hepatectomy., Methods: With over 1 000 ml of bleeding, 4 368 patients with hepatectomy between 1955 and 2000 were analysed retrospectively., Results: Among 4 368 patients receiving hepatectomy, 286 (6.5%) had massive hemorrhage because of damage to the major hepatic veins, portal hypertension, hepatic insufficiency, and the extensive adhesion around the tumor. Massive hemorrhage was managed by repair and transfixation of the damaged vessels; transfixation or devascularization of variceal bleeding; complete vessels ligation of the hepatic section with mattress suture; resection of the ruptured tumor after temporary occlusion of the porta hepatis; fibrinogen infusion; hot saline compression of the surface of the wound and/or daub biological glue; argon beam coagulation and packs placement., Conclusions: Light performance and nonforce dragging of liver can reduce massive hemorrhage caused by major vessel injury or tumor rupture. Normothetic occlusion of porta hepatis can reduce blood loss effectively when liver resection. In situ hepatectomy must be adopted if there is extensive adhesion around the tumor. Packs placement is still an effective measure to stop bleeding caused by defective coagulation and extensive blood oozing of wound surface.

In the past 37 years, 207 patients with hepatic injuries were managed in our hospital. Of them, 5 patients died during the preparation for emergency laparotomy. 191 patients were surgically treated, with 11 (5.8%) postoperative deaths, while all 11 patients treated nonsurgically were cured. Finally, we discussed operative methods of hepatic injuries and indications for conservative therapy.

By means of radioligand binding assay, peripheral-type benzodiazepine receptors (PBR) in the telencephalon, diencephalon, brain stem and cerebellum, were investigated in a rat model with hepatic encephalopathy (HE). Binding to PBR was studied using [3H]PK11195 as radioligand. It was demonstrated that only densities (Bmax) of [3H]PK11195 binding to crude synaptosomal membranes in the telencephalon, diencephalon of HE rats were obviously increased as compared with those of controls (P < 0.001 and P < 0.05 respectively). Increased affinities (Kd) were only observed in brain stem (P < 0.05). The results showed that the alterations of PBR in different regions of the brain play a significant role in the pathogenesis of HE.

In 40 patients with small hepatocellular carcinoma, the lesions were less than 5 cm in diameter, with a mean diameter of 3.4 +/- 1.1 cm. The smallest was 1.1 x 1.3 cm. 39 patients were first detected by ultrasonic scanning, and one was discovered during emergency operation because of spontaneous tumor rupture. 36 patients were treated by means of hepatic resection. One (2.7%) died within 30 days after operation. Four patients with severe hepatic cirrhosis and significant prolongation of prothrombin time (4 seconds over control) were subjected to selective transcatheter arterial embolization combined with transcatheter arterial infusion of chemotherapeutic agent. The diagnosis and treatment of small hepatocellular carcinoma was discussed.