Alternative management of anatomical right hemihepatectomy using ligation of inflow and outflow vessels without hilus dissection.

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<0.001) and portal triad occlusion (7 vs 11, P<0.001), blood loss (430 vs 580 mL, P=0.001), transfusion volume (300 vs 520 mL, P<0.001), 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, P<0.001), 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%, P<0.001) were found in the LCIOV group. CONCLUSION: The reported method is a safe and bloodless technique for right hemihepatectomy in select patients.

A new simple and safe technique of end-to-end invaginated pancreaticojejunostomy with transpancreatic U-sutures--early postoperative outcomes in consecutive 88 cases.

BACKGROUND: 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. MATERIAL AND METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Effects of mitofusin-2 gene on cell proliferation and chemotherapy sensitivity of MCF-7.

In order to evaluate the effect of mitofusin-2 gene (mfn2) on proliferation and chemotherapy sensitivity of human breast carcinoma cell line MCF-7 in vitro, pEGFPmfn2 plasmid carrying full length of mitofusin-2 gene was transfected, by using sofast, into MCF-7 cells. Mitofusin-2 gene expression in MCF-7 cells transfected by sofast after 48 h was detected by PCR and Western blotting, and the stable expression of GFP protein in MCF-7 cells by Western blot analysis. The proliferation of MCF-7 cells was assayed by MTT and cell counting. By using PI method, the effects of mfn2 on the cell cycle distribution of MCF-7 were measured. Annexin-V/PI double labeling method was employed to detect the changes in apoptosis induced by chemotherapeutics before and after transfection. The results showed that the MCF-7 cells transfected with mfn2 gene could stably and highly express GFP protein. MTT assay revealed that after transfection of mfn2 cDNA, the proliferation of MCF-7 cells was significantly inhibited. DNA histogram showed that cells arrested in S phase, and the percentage of S phase cells was 42.7, 17.2 and 19.6 in mfn2 cDNA transfection group, blank plasmid transfection group and blank control group, respectively (P<0.05). The apoptosis ratio of the cells transfected with mfn2 gene was increased from 3.56% to 15.95%, that of the cells treated with camptothecin (CAMP) followed by mfn2 gene transfection was 69.6%, and that in blank plasmid transfection group and blank control group was 31.0% and 23.4% respectively (P<0.05). It was suggested that transfection of mfn2 gene could significantly inhibit the proliferation of MCF-7 cells and promote their sensitivity to CAMP with a synergic effect.

[The technique of radical pancreaticoduodenectomy for malignant tumor in pancreatic head with pressed superior mesenteric blood vessels or portal vein].

OBJECTIVE: To investigate the technique of radical pancreaticoduodenectomy for malignant tumor in pancreatic head with pressed superior mesenteric blood vessel or portal vein. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

[Effect of mitofusin-2 gene in apoptosis of human breast carcinoma cell line in vitro].

OBJECTIVE: To investigate the role of mitofusin-2 gene (mfn2) in apoptosis in human breast carcinoma cell line MCF-7 cells after in vitro transfection. METHODS: 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. RESULTS: 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% (P < 0.05). Mfn2 gene induced break down and loss of mitochondrial cristae, and rarefaction of mitochondrial ground substance. Swollen mitochondria intensely aggregated around the cell nuclei. CONCLUSION: Mfn2 can strongly induce apoptosis in MCF-7 cells, which may be associated with decrease of mitochondrial transmembrane potential.

[Significances of gene differential expression patterns in hepatocirrhosis and non-hepatocirrhosis tissues within different ischemic time].

OBJECTIVE: To investigate the gene differential expression patterns in hepatocirrhosis and non-hepatocirrhosis tissues within different ischemic time. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Usefulness of oral mucosal epithelial cell apoptosis rate in nutritional assessment.

OBJECTIVE: This preliminary study was designed to explore a new method for nutritional assessment by measuring oral mucosal cell apoptosis or proliferation. METHODS: Forty-two consecutive patients with gastrointestinal malignant tumors were hospitalized on the surgical wards and studied prospectively. Patient-Generated Subjective Global Assessment was used to identify malnourished patients. Anthropometric measurements including weight, body mass index, triceps skinfold thickness, and midarm muscle circumference were recorded. The serum proteins measured were retinol-binding protein (RBP), transferrin, prealbumin (PA), and albumin. Simultaneously, the rates of oral epithelial cell apoptosis and proliferation were measured by flow cytometry. Of the 20 malnourished patients, 14 were followed up in a serial study with a 3-d nutritional support therapy. Nutritional indices and oral epithelial cell apoptosis rate were measured after 3 d of nutritional support. RESULTS: Malnutrition was diagnosed in 20 of 42 patients (47.6%). Oral epithelial apoptosis and proliferation rates were significantly decreased (P < 0.01 and P < 0.05, respectively) in malnourished compared with non-malnourished patients, although there were no significant differences between their anthropometric data. Malnourished patients had lower serum levels of RBP, albumin, and PA and rates of oral epithelial cell apoptosis and proliferation. The rate of oral epithelial cell apoptosis positively correlated with serum RBP (R = 0.32, P < 0.05) and PA (R = 0.33, P < 0.05). The rate of oral epithelial cell apoptosis and serum RBP and PA increased significantly in the malnourished patients who received nutritional support for 3 days. CONCLUSIONS: Measuring the rate of oral epithelial cell apoptosis may be another non-invasive technique to determine nutritional assessment and is worthy of further exploration.

Hepatectomy for huge hepatocellular carcinoma in 634 cases.

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.

Effects of 2-APB on store-operated Ca2+ channel currents of hepatocytes after hepatic ischemia/reperfusion injury in rats.

The effects of hepatic ischemia/reperfusion (1/R) injuries on hepatocellular viability and store-operated calcium current (Isoc) in isolated rat hepatocytes and the effects of 2-APB on store-operated calcium current (Isoc) in isolated rat hepatocytes after hepatic ischemia/reperfusion injuries were studied. Hepatic ischemia and reperfusion injury model was established and whole cell patch-clamp techniques were used to investigate the effects of 2-APB on Isoc. The results showed that ischemia/reperfusion injuries could significantly reduce hepatocellular viability and further increase Isoc in hepatocytes and 2-APB (20, 40, 60, 80, 100 micromol/L,) produced a concentration-dependent decrease of Isoc with IC50 value of 64. 63 +/- 10.56 micromol/L, (n = 8). It was concluded that ischemia/reperfusion injuries could reduce hepatocellular viability, probably through increased Isoc in hepatocytes and 2-APB had a protective effect on ischemia/reperfusion-induced liver injury, probably though inhibiting Isoc.

[Role of united hepatectomy and splenectomy in the surgical treatment of hepatocellular carcinoma complicated with hepatic cirrhosis and hypersplenism].

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.

[Study of immunological effect of dendritic cell transfected with survivin gene on the specific anti-alimentary tract tumor].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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 (P < 0.05). 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. CONCLUSIONS: 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.

Overcoming multi-drug resistance by anti-MDR1 ribozyme.

AIM: 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-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. RESULTS: The Rz- transfected HepG2 cells became doxorubicin-sensitive, concomitant with the decreases in MDR1 expression, P-gp amounts and efflux pump function. CONCLUSION: The approaches using either retrovirus or liposome-mediated transfer of anti-MDR1 ribozyme may be selectively applicable to the treatment of MDR cells.

Correlation of expression of multidrug resistance protein and messenger RNA with 99mTc-methoxyisobutyl isonitrile (MIBI) imaging in patients with hepatocellular carcinoma.

AIM: To explore whether P-glycoprotein (Pgp) and other pumps, multidrug resistance-associated protein (MRP) and lung resistance protein (LRP), could affect tumor accumulation and efflux of 99mTc-MIBI in liver cancer. METHODS: Surgically treated 78 liver cancer patients were included in this study. Before surgery, 99mTc-MIBI SPECT was performed 15 min and 120 min after injection of 20 mCi 99mTc-MIBI, respectively. Early uptake, delayed uptake (L/Nd), and washout rate (L/Nwr) of 99mTc-MIBI were obtained. Expressions of Pgp, MRP and LRP were investigated with Western blotting and immunohistochemistry. Messenger RNA (mRNA) level of Pgp, MRP and LRP was determined by RT-PCR. RESULTS: No 99mTc-MIBI uptakes in tumor lesions of 68 of 78 (87.2%) patients with hepatocellular carcinoma were found on 99mTc-MIBI SPECT. P-gp expression was observed in tumor tissues of the patients with no uptake of 99mTc-MIBI (P<0.017). No appreciable correlation was found between liver cancer 99mTc-MIBI images and expression of MRP or LRP on the level of protein or mRNA. CONCLUSION: 99mTc-MIBI SPECT is noninvasive, and useful in predicting the presence of MDR1 gene-encoded Pgp in patients with hepatocellular carcinoma.

Increased hepatic expression of nitric oxide synthase type II in cirrhotic rats.

AIM: To determine the role and effect of nitric oxide synthase type II (NOSII) in cirrhotic rats. METHODS: Expression of NOSII 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 NOSII is one of the important causes of hepatic cirrhosis and portal hypertension.

Heat shock protein 90 is responsible for hyperdynamic circulation in portal hypertensive rats.

AIM: To examine the participation of HSP90 in portal hypertensive rat mesentery in vitro. METHODS: 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. RESULTS: 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. CONCLUSION: HSP90 plays a key role in NO-dependent hyperdynamic circulation in portal hypertension and provides a novel method for future treatment of portal hypertension.

Construction of HCV-core gene vector and its expression in cholangiocarcinoma.

AIM: To establish an experimental model for exploring the role of hepatitis C virus (HCV) in the development of cholangiocarcinoma. METHODS: Recombinant plasmid of HCV-core gene was constructed with molecular cloning technique and transfected into QBC939 cells with lipofection.After it was selected with G418, resistant colonies were obtained. The colonies were analysed by immunocytochemistry and Western blotting. The morphology was observed under transmission electron microscope(TEM) and microscope. RESULTS: The recombinant plasmid was proved to carry the target gene by PCR and restriction enzymed mapping. Moreover, it could express HCV-C protein efficiently in QBC939 cells. The HCV-like particles were found in the cytoplasm by electron microscope, which were spherical with a diameter of 50 nm-80 nm possessing outer membrane. The transfected cells had lower differentiation and higher malignant degree under microscope. CONCLUSION: Because HCV-core gene could express steadily in cholangiocarcinoma cells,the transfected tumor cells(QBC939-HCVC) could be used to study the effect of HCV in the development of cholangiocarcinoma.

Expression of local renin and angiotensinogen mRNA in cirrhotic portal hypertensive patient.

AIM: To investigate the expression of local renin and angiotensinogen mRNA in cirrhotic portal hypertensive patients. METHODS: The expression of local renin and angiotensinogen mRNA in the liver, splenic artery and vein of PH patients was detected by RT-PCR analysis. RESULTS: Expression of local renin mRNA in the liver of control group was (0.19+/-0.11), significantly lower than that in splenic artery (0.45+/-0.17) or splenic vein(0.39+/-0.12) respectively, (P<0.05). Expression of local angiotensinogen mRNA in the liver was (0.64+/-0.21), significantly higher than that in splenic artery(0.41+/-0.15) or in splenic vein (0.35+/-0.18) respectively, (P<0.05). Expression of local renin mRNA in the liver, splenic artery and vein of PH group was (0.78+/-0.28), (0.86+/-0.35) and (0.81+/-0.22) respectively, significantly higher than that in the control group, (P<0.05). Expression of local angiotensinogen mRNA in the liver, splenic artery and vein of PH group was (0.96+/-0.25), (0.83+/-0.18) and (0.79+/-0.23) respectively, significantly higher than that in the control group, (P<0.05). There was no significant difference between the liver, splenic artery and vein in the expression of local renin or local angiotensinogen mRNA in PH group, (P<0.05). CONCLUSION: In normal subjects the expression of local renin and angiotensinogen mRNA was organ specific, but with increase of the expression of LRAS, the organ-specificity became lost in cirrhotic patients. LRAS may contribute to increased resistance of portal vein with liver and formation of splanchnic vasculopathy.

The immunotherapeutic effect of dendritic cells vaccine modified with interleukin-18 gene and tumor cell lysate on mice with pancreatic carcinoma.

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 immunotherapeutic 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-gamma were 2161+/-439 ng x L(-1) and 435+/-72 ng x 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 days in DC-Lysate groups, on 16 days in DC-IL18 groups, on 3 days in control group, but mice remained tumor-free for at least 50 days 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 days in DC-IL18-Lysate group. But the median survival was 48.6 days in DC-Lysate group, 33 days in DC-IL18 group, 17 days 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.

Effect of octreotide on human pancreatic cancer cells after transfected with somatostatin receptor type 2 gene.

AIM: 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. METHODS: 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). RESULTS: 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 (P<0.05) at the tested dosages (0.20, 0.40 and 0.80 microg/mL). CONCLUSION: 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.

Expression of nuclear factor kappa B in hepatitis C virus core gene transfected cholangiocarcinoma cells.

OBJECTIVE: To establish an experimental model for exploring the role of hepatitis C virus (HCV) in the development of cholangiocarcinoma. METHODS: Recombinant plasmids of HCV-C gene were constructed by molecular cloning techniques and identified by PCR and restriction enzyme mapping.The plasmids were then transfected into QBC939 cells (a cholangiocarcinoma cell line) by Lipofection. After selection with G418, resistant colonies were obtained and analyzed by immunocytochemistry and Western blotting. The morphology was observed by trans mission electron microscopy (TEM). The expression of NF-(k)B was detected by immunocytochemistry. RESULTS: Recombinant plasmid was shown by PCR and restriction enzyme mapping to carry the target gene. Moreover, it could efficiently express HCV-C protein in QBC939 cells. HCV-like particles were found in the cytoplasm by TEM, which were spherical with a diameter of 50-80 nm and possessed an outer membrane. Moreover, NF-(k)B activation could be shown in HCV core-transfected cells. CONCLUSION: Expression of the HCV-C gene in cholangiocarcinoma cells was achieved. Transfected tumor cells (QBC939-HCVc) could be used as a model to study the effect of HCV on the development of cholangiocarcinoma.