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Background and Aim: The conventional method of anatomical right hemihepatectomy (ARHH) requires hilus dissection. We report a method without hilus dissection to minimize intraoperative bleeding. Methods: We retrospectively evaluated data of 107 patients who received ARHH involving ligation of corresponding inflow and outflow vessels (LCIOV) without hilus dissection between January 2000 and October 2008. Results were compared to those of patients who underwent non-anatomical right hemihepatectomies (NARHH). Results: The two groups had similar gender and age (both, P > 0.05). The LCIOV group had a higher percentage of patients without intrahepatic metastases (94.6% vs 80.3%, P = 0.003). Hepatocellular carcinoma (HCC) lesion size (9.3 vs 10.2, P = 0.023), durations of inferior vena cava occlusion (4 vs 4.7, P

Mesohepatectomy is a rarely used operative procedure to treat liver cancer because of its technical complexity. In patients with hepatocellular carcinoma (HCC) with a viral hepatitic/cirrhotic background, this procedure can be used to resect the tumor with adequate margins, while at the same time preserve as much functioning hepatic parenchyma as possible. This retrospective study aimed to evaluate the early results and late survival outcomes of mesohepatectomy in HCC. From 1996 to 2005, 256 patients with HCC situated at the central liver segments (Couinaud segments IV, V, VIII ± I) were treated with mesohepatectomy. The treatment outcomes of these patients were retrospectively analyzed. The in-hospital mortality rate was 0.4%, but the postoperative morbidity rate was 28.1%. The 1-, 3-, and 5-year overall survival rates were 77.0, 49.8, and 35.1%, while the 1-, 3-, and 5-year disease-free survival rates were 59.1, 28.8, and 17.0%, respectively. Multivariate analyses showed the significant factors for overall survival were tumor size >8 cm, vascular invasion, and alpha fetoprotein (AFP) >5,000 ng/ml; and for disease-free survival were tumor size >8 cm, vascular invasion, tumor number (three or more), AFP >5,000 ng/ml. Mesohepatectomy is a safe and effective treatment for a centrally situated HCC with a viral hepatitic/cirrhotic background.

In treating hepatocellular carcinoma (HCC) patients with advanced cirrhosis, one of the most difficult problems is concomitant esophageal varices and hypersplenism. Whether these conditions should be treated surgically in association with HCC resection is still in debate. To elucidate whether esophageal devascularization or splenectomy is beneficial when simultaneously performed with liver resection in HCC patients with both varices and hypersplenism, HCC patients (n = 184) with esophageal varices and hypersplenism received one of the three treatments: simultaneous liver resection and esophageal devascularization (Group I, n = 41); simultaneous liver resection and splenectomy (Group II, n = 61); liver resection only (Group III, n = 82). The incidences of postoperative complications of the three groups were 31.7%, 29.5% and 24.4%, respectively, with no significant difference among them. The 5-year tumor-free survival rates for the group I, group II and group III were 34.1%, 36.1% and 37.8%, respectively. Variceal bleeding caused death by only 4.2% in group I, but by 14.3% in group II and 23.2% in group III. The survival rates in the group I and the group II were comparable to those in the group III, however, the recurrences of postoperative fatal variceal bleeding in group I and group II were significantly lower than those in group III. The results suggest that HCC patients with esophageal varices and hypersplenism should undergo hepatic resection plus esophageal devascularization or splenectomy if radical resection of HCC can be expected.

Background: The role of preoperative transcatheter arterial chemoembolization (TACE) for resectable hepatocellular carcinoma (HCC) was controversial. Methods: 246 patients with large centrally located HCC underwent mesohepatectomy (MH) and were divided into two groups: group A, 89 patients with preoperative TACE; group B, 157 patients without preoperative TACE. The aim was to evaluate the influence of preoperative TACE on postoperative complications and long-term results of patients with large centrally located HCC. Results: In the 89 patients of the TACE-MH group, a total of 123 (mean 1.4 per patient) preoperative TACEs were performed. The differences in postoperative complications (34.8 vs. 24.2%;p = 0.075) and overall hospital mortality (3.4 vs. 0.6%; p = 0.103) between the two groups were not significant. The postoperative recurrence rate in the remnant liver was higher in the MH group than in the TACE-MH group (79.6 vs. 73.0%), while the extrahepatic metastasis rate in the TACE-MH group was higher than that in the MH group (11.1 vs. 7.0%). Overall 1-, 3-, and 5-year survival rates were 87.1, 62.9, and 46.2%, respectively, for the TACE-MH group, and 82.2, 54.4, and 31.7%, respectively, for the MH group (p = 0.001); 1-, 3-, and 5-year disease-free survival rates were 75.0, 46.2, and 31.8%, respectively, for the TACE-MH group, and 69.6, 38.0, and 16.5%, respectively, for the MH group (p = 0.002). Conclusions: Long-term outcomes of patients with preoperative TACE were improved and the pattern of the recurrences after surgery was altered. The patients with large centrally located HCC could benefit more from this neoadjuvant treatment, although there was some influence of preoperative TACE on postoperative complications.

The role of surgical resection and thrombectomy for hepatocellular carcinoma (HCC) with portal vein tumor thrombus (PVTT) is controversial. This study aimed to evaluate the effects of the location and extent of PVTT on the long-term outcomes of surgical treatment for HCC. A total of 438 patients with HCC and PVTT underwent liver resection with or without thrombectomy. These 438 patients were divided into 2 groups: in group A, PVTT was located in the hepatic resection area or protruded into the first branch of the main portal vein beyond the resection line for

Objective To clarify the relationship between loss of expression of DPC4 proteins and pathogenesis of biliary tract carcinoma.

The conventional method of anatomical right hemihepatectomy (ARHH) requires hilus dissection. We report a method without hilus dissection to minimize intraoperative bleeding.We retrospectively evaluated data of 107 patients who received ARHH involving ligation of corresponding inflow and outflow vessels (LCIOV) without hilus dissection between January 2000 and October 2008. Results were compared to those of patients who underwent non-anatomical right hemihepatectomies (NARHH).The two groups had similar gender and age (both, P0.05). The LCIOV group had a higher percentage of patients without intrahepatic metastases (94.6% vs 80.3%, P=0.003). Hepatocellular carcinoma (HCC) lesion size (9.3 vs 10.2, P=0.023), durations of inferior vena cava occlusion (4 vs 4.7, P0.001) and portal triad occlusion (7 vs 11, P0.001), blood loss (430 vs 580 mL, P=0.001), transfusion volume (300 vs 520 mL, P0.001), and measures of postoperative liver function (e.g. maximum aspartate aminotransferase [AST]) of the LCIOV group were also significantly less than the NARHH group. Larger hepatic cavernous hemangiomas (HCH) lesion size (16.2 vs 13.0, P0.001), longer operative time (168 vs 154 min, P=0.017), and a lower percentage of patients with inferior vena cava occlusion (17.8% vs 35.2%, P=0.001), pleural effusions (19.3% vs 30.9%, P=0.042), and blood transfusions (10.3% vs 75.0%, P0.001) were found in the LCIOV group.The reported method is a safe and bloodless technique for right hemihepatectomy in select patients.

Postoperative pancreatic fistula (POPF) remains a leading cause of morbidity and mortality after pancreaticoduodenectomy (PD). Thus, a number of technical modifications regarding the pancreato-enteric anastomosis after PD have been proposed to reduce POPF rate. Until now, there is no consensus on which is the best. This study presents a new technique of the end-to-end invaginated pancreaticojejunostomy with two to three transpancreatic U-sutures and evaluates its safety and reliability.From 2002 to 2007, 88 patients (54 men and 34 women) underwent an invaginated end-to-end pancreaticojejunostomy with two to three transpancreatic U-sutures after PD. The mean age was 52.4 years (range, 26-74 years). The diseases of the all patients were malignant.In all patients of this study, two transpancreatic U-sutures were performed in 59 and three U-sutures in 29. The median duration of surgery was 3.8 h (range 3-6.5) and the median time to perform pancreaticojejunostomy was 13.3 min (range 8-25). The median blood loss was 750 ml (range 300-1,800), 36 patients needed transfusion and the median blood transfusion was 380 mL (range 200-1,200). Overall morbidity occurred in 15 patients (17.0%). Only two patients (2.2%) had grade A of POPF and no patient had grade B and grade C of POPF. No operative death occurred.An invaginated end-to-end pancreaticojejunostomy with two to three transpancreatic U-sutures is simple, rapid, safe, and reliable technique, even in some patients with soft pancreas and small pancreatic duct.

To investigate the technique of radical pancreaticoduodenectomy for malignant tumor in pancreatic head with pressed superior mesenteric blood vessel or portal vein.From March 2005 to March 2007, thin slice scan and vessel-reconstruction of 56 patients of malignant tumor in pancreatic head with pressed superior mesenteric blood vessels or portal vein were carried out using multidetector spiral CT to evaluate whether peripheral vessels of pancreatic tumor were invaded and whether the tumor was resectable. During the operation, 3 vascular blocking bands for superior mesenteric vein, portal vein and spleen vein or 4 vascular blocking bands (additional one for inferior mesenteric vein) were preset. Under the cross and traction between superior mesenteric vein and superior mesenteric artery, resected the uncinate process of pancreas thoroughly. Using those methods, radical pancreaticoduodenectomy for 56 patients above-mentioned were successfully accomplished.The accuracy for preoperative judging by using multidetector spiral CT whether the peripheral vessels of pancreatic cancer were invaded and whether the tumor was resectable was 98% and 100% separately. Thirty-seven of 56 patients, whose superior mesenteric blood vessels or portal veins were pressed by the tumor of pancreatic head, were operated using 3 vascular blocking bands and 2 patients using 4 vascular blocking bands, followed by suturing the bleeding points of the superior mesenteric vein with 5-0 vascular suture Proline. One patient's superior mesenteric vein was partially resected and restored. The operations cost 5-8 h each and the blood loss was 200-600 ml. There were no operative or postoperative hemorrhage or pancreatic juice leakage. According to the follow-up up to now, 2 patients died of multiple live tumor metastases 7 and 9 months separately after operation, the other 54 patients were still alive.Thin slice scan and vessel-reconstruction using multidetector spiral CT can accurately judge whether the blood vessels near the pancreatic tumor were invaded and whether the tumor was resectable, using 3 vascular blocking bands or 4 vascular blocking bands and cross, traction of the superior mesenteric blood vessels, operator can easily accomplish the radical pancreaticoduodenectomy of malignant tumor in pancreatic head with pressed superior mesenteric blood vessels and portal vein, which was not resectable or need combined resection of the blood vessels in the traditional opinion.

To investigate the role of mitofusin-2 gene (mfn2) in apoptosis in human breast carcinoma cell line MCF-7 cells after in vitro transfection.pEGFP mfn2 was transfected by sofast in vitro. Expression of GFP was observed by Western blot, and the MCF-7 cell proliferation was measured by MTT and cell counting. Apoptosis in MCF-7 cells was observed in annexin-V/PI and chondrosome transmembrane potential of MCF-7 marked in JC-1 by FCM. The Ultrastructure of cells was observed by transmission electron microscopy.The stable expression of GFP in MCF-7 cells was confirmed by Western blot. Mfn2 significantly inhibited cell proliferation, revealed by MTT, and decrease chondrosome transmembrane potential. Exogenous mfn2 gene significantly induced apoptosis. The apoptotic rate was increased from 3.6% to 16.0% (P0.05). Mfn2 gene induced break down and loss of mitochondrial cristae, and rarefaction of mitochondrial ground substance. Swollen mitochondria intensely aggregated around the cell nuclei.Mfn2 can strongly induce apoptosis in MCF-7 cells, which may be associated with decrease of mitochondrial transmembrane potential.

Background The key to Belghiti's liver-hanging maneuver is to develop a retrohepatic tunnel. This procedure requires a blind dissection of the plane anterior to the inferior vena cava (IVC), with the inherent risks of damaging the short hepatic veins and consequential bleeding. The aim of this article is to describe a liver double-hanging maneuver with the advantage of being technically simple and safe. Methods The operator uses his or her right index finger to dissect the space from below upward between the hepatic parenchyma and the anterior and superior edge of the right adrenal gland, which is situated just on the right side of the IVC. The operator then uses his left index finger to dissect the retrohepatic space from above downward on the right side of suprahepatic IVC, which is lateral to where the right hepatic vein joins the IVC. The retrohepatic tunnel is built when the 2 fingers touch each other. A kidney pedicle forceps is used to place 2 tapes around the liver for suspension. Results In all, 65 patients underwent right hepatectomy using this maneuver. The study included 62 patients with hepatocellular carcinoma (tumor size: mean ± SD, 10 ± 3.7 cm), and 3 patients had hepatic cavernous hemangioma, with a maximum diameter of 12.6 cm, 14.4 cm, and 22.6 cm, respectively. No major bleeding was encountered during the creation of the retrohepatic tunnel, with a success rate of 100%. Conclusion To develop the retrohepatic tunnel in the space on the right of the IVC is absolutely bloodless, and it is technically easy and safe.

Massive bleeding remains one of main factors of morbidity and death in liver resections. For this reason, the Pringle maneuver or total vascular exclusion is commonly used during liver resection. However, ischemic damage is still a major problem. Some surgeons used the "glissonean" approach for bleeding control, but the technique is tedious and also time consuming, with high incidence of bile leaks in the postoperative period. The aim of this paper is to describe a new bleeding control technique, rapid ligation of the corresponding inflow and outflow vessels without hilus dissection before the parenchyma transection during anatomical left liver resection and to analyze the feasibility, blood loss, transfusion requirements, and postoperative complications.During the past 18 years, we used the new hemorrhage control technique in left liver resection in 630 patients with malignant or benign tumors.The median blood loss in all 630 patients was 110 +/- 250 ml (range 50-750), and no patient required blood transfusion. The median total operative time was 77 +/- 35 min (range 25-155). No bile leaks and liver failure of the patients occurred postoperatively. There was no death within 30 postoperative days.The rapid ligation of the corresponding inflow and outflow vessels without hilus dissection before the parenchyma transection is a feasible, safe, and bloodless technique during the left liver resection.

To investigate the gene differential expression patterns in hepatocirrhosis and non-hepatocirrhosis tissues within different ischemic time.The liver tissues were divided into two groups: Group A (non-hepatocirrhosis), Group B (hepatocirrhosis), each of which consisted of 3 groups with different ischemic time: 15, 30 and 45 minutes. The gene differential expression patterns in the two groups within different ischemic time were detected and compared with those in normal liver tissues by using 4000 points gene microarray.In non-hepatocirrhosis tissues, the homeostatic maintenance genes expressed highly during hepatic ischemia for 15 minutes, and no apoptotic gene was expressed; but in hepatocirrhosis tissues, many apoptotic genes expressed highly. As for 30 minutes, in both two groups liver tissue genes expressed to the peak, and the genes related to cell death, oxidative stress and nuclear factors expressed highly. The difference lies in the facts that in Group B pro-apoptosis genes expressed more than those in Group A, and the Ratio values were higher than those in Group A. Many genes of heat shock protein family and antioxidant proteins expressed highly simultaneously in Group A, but comparatively low in Group B. As for 45 minutes, genes of heat shock proteins and antioxidant proteins expressed lowly in Group B.It suggests that the safe time limit of hepatic ischemia for cell survive is 30 minutes or so. Non-hepatocirrhosis tissues could endure 30 minutes of ischemia and even longer, but it should be restricted within 30 minutes in hepatocirrhosis tissues.

To investigate the role of united hepatectomy and splenectomy in the surgical treatment of hepatocellular carcinoma complicated with hepatic cirrhosis and hypersplenism.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.(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)%, P0.05]; 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 (P0.05). (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%, P0.05), but the survival rate without tumor of group A was 37.7%, higher than that of group B (18.9%, P0.05).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.

To investigate the effects of dendritic cells (DCs) transfected with survivin gene, and to observe the effective and specific anti-tumor immunological effect induced by modified DC in vitro.Survivin gene was transfected to DCs with liposomes. Survivin expression could be detected both in DCs cells and in cell culture with method of Western blot. Cytokines as well as cellular surface molecule such as IL-12, TNF-alpha, CD1 alpha, CD83, MHCII, CD80 and CD86 were detected. The competence of inducing human specific cytotoxic T lymphocyte (CTLs) was also detected with MTT.Survivin expression could be detected both in DCs which were transfected with survivin cDNA and in cell culture superior. The IL-12 and TNF-alpha level was (265.2 +/- 32.7), (437.1 +/- 83.5) pg/ml, and much higher in transgened DC cells than blank DC cells (P0.05). CD1 alpha, CD83, MHCII, CD80 and CD86 was high expressed in survivin-DC cells, however, it was low expressed in blank DC cells. The lyse rate to gastric cancer cell, colon cancer cell and bile duct cancer cell was 65%, 77%, and 85% respectively, and these were much higher than those of blank DC cells.DCs transfected with survivin gene could induce specific cytotoxic T lymphocytes and strikingly raised DC cell's antigen present function, and have specific CTL killing activity.

To explore the relationship between kruppel-like factor (KLF)6 gene and the development or progression of hepatocellular carcinoma (HCC).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.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, P0.05). 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.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.

To observe the effect of octreotide on apoptosis rate of human pancreatic cancer cells PC-3 after transfected with somatostatin receptor type 2 (SST2) gene.SST2 plasmid was transfected into PC-3 cells by liposome. Result of transfection was detected by immunocytochemical staining and Western blotting. Apoptosis rates of PC-3 cells under different dosages of octreotide were measured by MTT assay and flow cytometry (FCM).Apoptosis rate caused by octreotide of transfected PC-3 cells was 7.56+/-1.06% at the dosage of 0.20 microg/mL, 9.25+/-1.73% at the dosage of 0.40 microg/mL and 14.18+/-2.71% at the dosage of 0.80 microg/mL. Apoptosis rate caused by octreotide of non-transfected PC-3 cells was 5.76+/-0.75% at the dosage of 0.20 microg/mL, 6.69+/-0.80% at the dosage of 0.40 microg/mL and 7.26+/-1.28% at the dosage of 0.80 microg/mL. Transfected PC-3 cells growth inhibition rate caused by octreotide was 9.36+/-1.34% at the dosage of 0.20 microg/mL, 12.03+/-1.44% at the dosage of 0.40 microg/mL and 20.23+/-4.21% at the dosage of 0.80 microg/mL. Non-transfected PC-3 cells growth inhibition rate caused by octreotide was 6.44+/-0.66% at the dosage of 0.20 microg/mL, 7.65+/-0.88% at the dosage of 0.40 microg/mL and 9.29+/-1.32% at the dosage of 0.80 microg/mL. We found that octreotide caused higher apoptosis rate and inhibition rate in transfected groups than in non-transfected groups (P0.05) at the tested dosages (0.20, 0.40 and 0.80 microg/mL).Deficiency of SST2 was probably the major reason why octreotide had little effect on PC-3 cells. Transfecting SST2 gene could strengthen the ability of octreotide of killing PC-3 cells. It provided an experimental evidence for using both octreotide and transfection with SST2 gene on clinical treatment of pancreatic cancer.

To transfect antisense vector of human cyclooxygenase-2 (COX-2) gene into COX-2 highly expressing cholangiocarcinoma cell line QBC939 and explore its biological activities and role in carcinogenesis.QBC939 cells were transfected with antisense vector of human COX-2 gene using LipoVec transfecting technique. Transfected cells were selected with G418; COX-2 mRNA was examined using reverse transcription polymerase chain reaction (RT-PCR) and COX-2 protein expression was detected by immunocytochemistry using isozyme selective antibodies. The proliferative status of transfected cells was measured by using methabenzthiazuron (MTT) assay; Cell cycle and apoptosis were analyzed by using flow cytometry.RT-PCR showed a lower COX-2 mRNA level in antisense vector transfected cells and immunocytochemistry showed a weaker COX-2 protein expression in antisense vector transfected cells. The antisense vector transfected cells proliferative index decreased significantly (P0.01), the percentage of S phase decreased remarkably (P0.05) in antisense vector transfected cells (9.27% +/- 1.91%) compared with that in QBC939 cells without transfection(16.35% +/- 2.87%), and the percentage of G0/G1 phase increased remarkably (P0.05) in antisense vector transfected cells (75.16% +/- 4.13%) compared with that in QBC939 cells without transfection (57.31% +/- 10.16%). Transfection with antisense vector of human COX-2 gene had no significant influence on the apoptosis in QBC939 cells (P0.05).Transfection with antisense vector of human COX-2 gene could inhibit the proliferation of human cholangiocarcinoma QBC939 cells.

To reverse multidrug resistance (MDR) of HepG2 by anti-MDR1 hammerhead ribozyme.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.The Rz-tranduced HepG2 cells became doxorubicin-sensitive, concomitant with the decreases in MDR1 expression, P-gp amounts and efflux pump function.The approaches using either retrovirus or liposome-mediated transfer of anti-MDR1 ribozyme may be selectively applicable to the treatment of MDR cells.

To investigate the effects of HBx gene on proliferation activity of hepatoma cells in vitro and in vivo.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.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.HBx gene can facilitate the proliferation of hepatoma cells both in vitro and in vivo.

To study the role of heat shock protein 90 (HSP 90) in the formation of hyperdynamic circulation of portal hypertensive rats.Animal model of portal hypertension was established by partial ligation of the portal vein among 20 SD rats (group P). Ten rats underwent sham operation (group S) Four weeks after the operation, the rats were killed and their mesenteric arteries and portal veins were obtained. The expression of HSP 90 in the portal veins and part of the superior mesenteric arteries was detected by Western blotting. Another mesenteric arteries underwent continuous perfusion with methoxamine (MTX, 30 and 100 micro mol/L) or acetylcholine (Ach). The perfusion pressure was monitored by a sensor. The concentration-response curves were examined in response to MTX and Ach infusion respectively. Then geldanamycin (GA) was infused; responses to MTX and ACh were repeated as described above. The endothelia of these arteries was denuded. After endothelial denudation was achieved, response to sodium nitroprusside (SNP) was examined.The expression of HSP 90 was much stronger in the group P than in the group S. The perfusion pressure of the mesenteric artery of the group P was 13.32 mm Hg +/- 0.55 mm Hg, significantly lower than that of the group S (17.33 mm Hg +/- 0.57 mm Hg, n = 10, P0.001). After the addition of MTX, the perfusion pressure of the mesenteric artery of the group P was 141.9 mm Hg +/- 7.6 mm Hg, significantly lower than that of the group S (181.1 mm Hg +/- 16.2 mm Hg, n = 8, P0.001). GA and 30 micro mol/L or 100 micro mol/L MTX were added successively into the perfusion fluid for the mesenteric superior artery of the group P, then the perfusion pressure became 106.7 mm Hg +/- 7.5 mm Hg or 124.9 mm Hg +/- 5.5 mm Hg, significantly higher than those of the mesenteric superior artery of the group P without addition of GA (60.9 mm Hg +/- 4.3 mm Hg, n = 5, P0.05 and 98.7 mm Hg +/- 4.7 mm Hg, n = 5, P0.05). GA and 100 micro mol/L MTX were added successively into the perfusion fluid for the mesenteric superior artery of the group S, the perfusion pressure became 151.2 mm Hg +/- 7.1 mm Hg, not significantly different from that of the mesenteric superior artery of the group S without addition of GA (149.8 mm Hg +/- 5.6 mm Hg, P0.05). With the addition of GA in advance, the ACh-dependent vasorelaxation of the isolated perfused rat mesenteric artery was significantly attenuated in comparison with the mesenteric artery without addition of GA into whose perfusion fluid (P0.05). However, GA did not affect the vasodilation of the ensothelium-neduded mesenteric artery in response to SNP.HSP90 is responsible for the hyperdynamic circulation in the portal hypertensive rats.

To explore the objective basis of the time and characteristics of refeeding of patients with acute pancreatitis (AP).AP patients were randomly divided into routine group (n=105) and special group (n=99). The refeeding time and characteristics of the routine group were based on their levels of blood and urine amylase, function of the gastrointestinal tract, and symptoms and signs, and those of the special group on their imaging changes of ultrasonography and CT of the pancreas. Clinical data from the groups were analyzed prospectively and statistically.At the beginning of refeeding, patients of the two groups showed a high recurrence rate of abdominal pain (routine group, 9.1%; special group, 10.5%), but no AP relapse. Three days to 2 weeks after refeeding, the patients of the routine group had a higher recurrence rate of abdominal pain (11.1%) and a relapse rate of AP(14.1%) as compared with those of the special group (P0.05). Two weeks later, both rates decreased apparently. However, 3 days to 4 weeks after refeeding, the patients of the special group had no recurrence of abdominal pain and relapse of AP. Imaging changes of the pancreas and peripancreatic tissue were not consistent with the symptoms and signs of AP patients. The higher the Balthazar CT grading and APACHE-II score, the higher the recurrence rate of abdominal pain and the relapse rate of AP after refeeding (P0.05).Symptoms and signs were usually discordant to the imaging changes of the pancreas in AP patients. Imaging changes of the pancreas might serve as a basis for the best occasion of refeeding light semi-fluid or light food in AP patients.

To clarify the relationship between loss of DPC4 gene expression and pathogenesis of pancreatobiliary carcinoma.75 slides of normal duct (20), hyperplasia (15), dysplasia (15), invasive carcinoma (25) from patients with pancreatic diseases including pancreatic carcinoma (25 patients), chronic pancreatitis (6), pancreas injury (2) and 71 slides of common bile duct (CBD) carcinoma (38), gallbladder carcinoma (18), hilar bile duct (HBD) carcinoma (15) from patients with primary biliary tract carcinoma were analyzed for the expression of DPC4 protein by immunohistochemical staining.All specimens from 20 cases of normal duct and 15 cases of hyperplasia showed marked expression of DPC4 protein. The frequency of loss expression of the DPC4 gene was 33% in dysplasia, and 48% in invasive carcinoma. There was a significant statistical difference between hyperplasia and dysplasia (P0.01) and in dysplasia vs invasive carcinoma (P0.05). The frequency of loss expression of the DPC4 gene was 47.3% in CBD carcinoma, 11% in gallbladder carcinoma, and 13% in HBD carcinoma. The frequency of loss expression of the DPC4 gene was significantly different in CBD carcinoma vs gallbladder carcinoma and HBD carcinoma (P0.01).Inactivation of the DPC4 gene occurs late in the neoplastic progression of pancreatic carcinoma. The frequency of DPC4 gene alternation was different in various locations of biliary tract carcinoma. In CBD carcinoma, this frequency is similar to that in pancreatic carcinoma, indicating their similar molecular alternations.

To discuss the safety and feasibility of hepatectomy for huge primary liver cancer (PLC).The effect of resection of huge PLC was examined retrospectively. Some problems in resection of huge PLC were discussed.Of 375 patients with huge PLC undergoing hepatectomy, 11 (2.9%) died in one month after operation. The 1-, 2-, 3-, 5-and 10-year survival rates of the patients were 63.3%, 45.6%, 34.7%, 16.5% and 1.8%, respectively. The effect of prolonging survival time was significant.Hepatectomy for huge PLC is safe, feasible, and effective.

To study the correlation between human cholangiocarcinoma in the porta hepatis and the infection of hepatitis virus.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.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.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.

To determine the effect of Salviae miltiorrhizae on cirrhosis and portal hypertension by inhibiting nitric oxide synthase type II (NOSII) in rats.Real time RT-PCR was used to detect the expression of NOSII mRNA. The enzymatic activity of nitric oxide synthase and the circulating levels of nitric oxide (NO), systemic and portal hemodynamics, and quantification of cirrhosis were measured with highly sensitive methods. Traditional Chinese medicine was utilized to treat cirrhotic rats and the function of NO was evaluated. Double-blind method was applied in the experiment constantly.The concentration of NO increased markedly at all stages of cirrhosis, and so did the enzymatic activity of NOS, and the iNOSmRNA expressed greatly. Meanwhile the portal-venous-pressure (PVP), portal-venous-flow (PVF) increased significantly. NO, NOS and iNOSmRNA were positively correlated to the quantity of hepatic fibrosis. Salviae miltiorrhizae significantly inhibited NO production and inhibited the expression of iNOSmRNA.The increased hepatic expression of NOSII is one of the important factors causing cirrhosis and portal hypertension. Salviae miltiorrhizae significantly ameliorates cirrhosis and portal hypertension.

To compare curative effects of various surgical procedures of bile duct stones.Two thousand nine hundred and fifty-five patients with intrahepatic bile duct lithiasis who had undergone various surgical procedures were analysed with Meta-analysis. Some of these cases were reported in Chinese Medical Journals from January 1990 to March 2001 and others were from Tongji Hospital.There was a significant difference between curative effects of non-hepatectomy and that of hepatectomy (chi(2) = 62.945, P0.01), and the outcomes of hepatectomy were much better than those of non-hepatectomy with OR(S) equalled to 0.303 (0.222 - 0.413). There was not a significant difference between curative effect of interposed jejunum and that of hepatectomy (95% CI of RR from 0.98 to 1.04). All the other operation, effects were worse than hepatectomy (upper limit of 95% CI of RR1).Hepatectomy is the most ideal surgery for intrahepatic bile duct stones and operation methods should be diversified since good effect could also be obtained when other operations are performed on suitable cases.

To analyse the causes and the management of massive hemorrhage in hepatectomy.With over 1 000 ml of bleeding, 4 368 patients with hepatectomy between 1955 and 2000 were analysed retrospectively.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.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.

To reverse multidrug resistance (MDR) of HepG2 by anti-MDR1 hammerhead ribozyme.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-transfected 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-transfected cells, and Rhodamine123 (Rh123) applied to test the function of Pgp.The Rz- transfected HepG2 cells became doxorubicin-sensitive, concomitant with the decreases in MDR1 expression, P-gp amounts and efflux pump function.The approaches using either retrovirus or liposome-mediated transfer of anti-MDR1 ribozyme may be selectively applicable to the treatment of MDR cells.

AIM: To clarify the safety and feasibility of hepatectomy for huge hepatocellular carcinoma (HCC). METHODS: A total of 4765 patients with HCC operated at Tongji Hospital were retrospectively studied, of them, 780 patients had huge HCC (10 cm or more in diameter). Hepatectomy was carried out on 634 patients (81.2%). The majority of the liver resection were major resections, and combined resection of the adjacent organs or structures was common (17.2%). The liver resection was combined with portal vein thrombectomy in 139 patients (21.9%). RESULTS: Postoperative complications were common (26.8%) and required another laparotomy to prevent the complications in 5 patients (0.8%). The 30-d mortality was 2.2%. The main causes of postoperative deaths were liver failure (n = 9), postoperative bleeding (n = 4) and septic complication (n = 1). The 3-, 5- and 10-year survival rates after liver resection were 35.1%, 18.2% and 3.5%, respectively. CONCLUSION: Hepatectomy for huge HCC is safe and effective. It should be used to treat patients with low surgical risks and resectable tumours.

AIM: To determine the role and effect of nitric oxide synthase type II (NOS II) in cirrhotic rats. METHODS: Expression of NOS II mRNA was detected by real time RT-PCR. The activity of nitric oxide synthase and serum levels of NO, systemic and portal hemodynamics and degrees of cirrhosis were measured with high sensitive methods. Chinese traditional medicine tetrandrine was used to treat cirrhotic rats and to evaluate the function of NO. Double-blind method was applied during the experiment. RESULTS: The concentration of NO and the activity of NOS were increased markedly at all stages of cirrhosis, and iNOSmRNA was greatly expressed. Meanwhile the portal-venous-pressure (PVP), and portal-venous-flow (PVF) were significantly increased. NO, NOS and iNOSmRNA were positively correlated to the quantity of hepatic fibrosis. Tetrandrine significantly inhibited NO production and the expression of iNOSmRNA. CONCLUSION: Increased hepatic expression of NOS II is one of the important causes of hepatic cirrhosis and portal hypertension.

To examine the participation of HSP90 in portal hypertensive rat mesentery in vitro.Immunohistochemistry and Western-blot were used to examine the expression of HSP90 in mesenteric vasculature. HSP90 mRNA was detected by RT-PCR, and the role of HSP90 in hyperdynamic circulation was examined by in vitro mesenteric perfusion studies.HSP90 was overexpressed in endothelium of mesentery vasculature in animals with experimental portal hypertension induced by partial portal vein ligation (PVL) compared with normal animals. Geldanamycin (GA), a special inhibitor of HSP90 signaling, attenuated ACh-dependent vasodilation but did not affect vasodilation in response to sodium nitroprusside in normal rats. In PVL animals, the perfused mesentery was hyporesponsive to vasoconstrictor methoxamine. GA significantly potentiated methoxamine-induced vasoconstrictor after PVL.HSP90 plays a key role in NO-dependent hyperdynamic circulation in portal hypertension and provides a novel method for future treatment of portal hypertension.

AIM: To estimate the effect of a therapeutic vaccine against pancreatic carcinoma based on dendritic cell (DC) vaccine modified with tumor lysate and Interleukin-18 gene. METHODS: The BALB/C mice model of pancreatic carcinoma was induced with DMBA. DC vaccine was constructed through pulsed with tumor lysate and transfected by the recombinant adenoviral vector encoding IL-18 gene. The immnotherapeutic effects of DC vaccine on mice with pancreatic carcinoma were assessed (divided into DC-IL18-Lysate group, DC-Lysate group, DC-IL18 group, DC group, PBS group). RESULTS: After vaccination of the DC vaccine, the concentration of IL-18 and IFN-γ were 2161 ± 439 ng·L-1 and 435 ± 72 ng·L-1 in DC-IL18-Lysate group and there was significant difference compared with other groups (P < 0.01). After vaccination of the DC vaccine, the transplanted tumors were observed on 30 d in DC-Lysate groups, on 16 d in DC-IL18 groups, on 3 d in control group, but mice remained tumor-free for at least 50 d in DC-IL18-Lysate group and there was significant difference between DC-IL18-Lysate group and other groups (P < 0.01). The median survival exceeds 62 d in DC-IL18-Lysate group. But the median survival was 48.6 d in DC-Lysate group, 33 d in DC-IL18 group, 17 d in PBS group. The survival period was obviously prolonged in DC-IL18-Lysate group than in other groups (P < 0.05, P < 0.01). The weight of pancreatic tumor was 0.22 ± 0.083 g in DC-IL18-Lysate group, 1.45 ± 0.74 g in DC-Lysate group, 1.89 ± 1.34 g in DC-IL18 group, 3.0 ± 1.6 g in DC group, 2.9 ± 2.0 g in PBS group and the weight of tumor obviously reduced in DC-IL18-Lysate group than in other groups (P < 0.05, P < 0.01). CONCLUSION: DC vaccine modified with tumor lysate and Interleukin-18 gene can induce a specific and effective immune response against pancreatic carcinoma cell.

AIM:To determine the site of production and uptake of tumor necrotic factor alpha (TNFalpha),and evaluate the relationship between serum TNFalphaand pla-sma endotoxin (ET) in rats with acute hemorrhagic necrotic pancreatitis (AHNP).METHODS:Sprague Dawley rats were divided into AHNP group and control group (n = 12). AHNP model was induced by retrograde injection of 5% sodium taurocholate via pancreatic bile duct. The blood samples were obtained through portal vein 2 and 6 hours after the operation.RESULTS:The contents of TNFalphain portal vein were increased rapidly in the development of AHNP. They were lower in hepatic vein (280.59 plus minus 20.02) and femoral artery (310.82 plus minus 7.97) than in portal vein (354.91 plus minus 25.50) (P < 0.05), and higher in femoral artery than in hepatic vein 6 hours after the operation (P