Minimally Invasive Surgical Procedures ; Philosophy, Medical

OBJECTIVETo reverse multidrug resistance (MDR) of HepG2 by anti-MDR1 hammerhead ribozyme.

METHODSWe 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.

RESULTSThe Rz-tranduced HepG2 cells became doxorubicin-sensitive, concomitant with the decreases in MDR1 expression, P-gp amounts and efflux pump function.

CONCLUSIONSThe approaches using either retrovirus or liposome-mediated transfer of anti-MDR1 ribozyme may be selectively applicable to the treatment of MDR cells.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Cell Line, Tumor ; Drug Resistance, Multiple ; genetics ; Genetic Vectors ; genetics ; Humans ; RNA, Catalytic ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

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

METHODSReverse-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.

RESULTSAn 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.

CONCLUSIONWe 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.

Carcinoma, Hepatocellular ; genetics ; DNA ; genetics ; DNA Mutational Analysis ; Humans ; Kruppel-Like Transcription Factors ; genetics ; Liver Neoplasms ; genetics ; Mutation ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, RNA

OBJECTIVETo 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.

METHODSQBC939 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.

RESULTSRT-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 (P < 0.01), the percentage of S phase decreased remarkably (P < 0.05) 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 (P < 0.05) 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 (P > 0.05).

CONCLUSIONTransfection with antisense vector of human COX-2 gene could inhibit the proliferation of human cholangiocarcinoma QBC939 cells.

Apoptosis ; Bile Duct Neoplasms ; metabolism ; pathology ; Bile Ducts, Intrahepatic ; Cell Cycle ; Cell Division ; Cell Line, Tumor ; Cholangiocarcinoma ; metabolism ; pathology ; Cyclooxygenase 2 ; DNA, Antisense ; genetics ; Humans ; Isoenzymes ; biosynthesis ; genetics ; physiology ; Membrane Proteins ; Prostaglandin-Endoperoxide Synthases ; biosynthesis ; genetics ; physiology ; RNA, Messenger ; genetics ; Transfection

Animals ; Carcinoma, Hepatocellular ; blood supply ; virology ; Cell Transformation, Neoplastic ; Endothelial Growth Factors ; biosynthesis ; genetics ; Gene Expression Regulation, Viral ; Hepatitis B virus ; pathogenicity ; physiology ; Humans ; Liver Neoplasms ; blood supply ; virology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Trans-Activators ; physiology

OBJECTIVETo investigate the role of united hepatectomy and splenectomy in the surgical treatment of hepatocellular carcinoma complicated with hepatic cirrhosis and hypersplenism.

METHODSTwo 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).

CONCLUSIONSThe 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.

Adult ; Carcinoma, Hepatocellular ; complications ; immunology ; mortality ; surgery ; Female ; Hepatectomy ; Humans ; Hypersplenism ; complications ; surgery ; Liver Cirrhosis ; complications ; surgery ; Liver Neoplasms ; complications ; immunology ; mortality ; surgery ; Male ; Middle Aged ; Prospective Studies ; Splenectomy ; Survival Rate ; Treatment Outcome

OBJECTIVETo 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.

METHODSSurvivin 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.

RESULTSSurvivin 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.

CONCLUSIONSDCs 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.

Antigens, CD ; metabolism ; Dendritic Cells ; immunology ; Gastrointestinal Neoplasms ; therapy ; Humans ; Immunotherapy, Active ; In Vitro Techniques ; Inhibitor of Apoptosis Proteins ; Interleukin-12 ; secretion ; Microtubule-Associated Proteins ; genetics ; Neoplasm Proteins ; genetics ; Transfection ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo investigate the technique of radical pancreaticoduodenectomy for malignant tumor in pancreatic head with pressed superior mesenteric blood vessel or portal vein.

METHODSFrom 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.

RESULTSThe 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.

CONCLUSIONSThin 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.

Adult ; Aged ; Female ; Humans ; Male ; Mesenteric Artery, Superior ; pathology ; surgery ; Mesenteric Veins ; pathology ; surgery ; Middle Aged ; Neoplasm Invasiveness ; Pancreas ; pathology ; Pancreatic Neoplasms ; pathology ; surgery ; Pancreaticoduodenectomy ; methods ; Portal Vein ; pathology

OBJECTIVETo investigate the effects of HBx gene on proliferation activity of hepatoma cells in vitro and in vivo.

METHODSThe 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.

RESULTSThe 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.

CONCLUSIONHBx gene can facilitate the proliferation of hepatoma cells both in vitro and in vivo.

Animals ; Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Cycle ; genetics ; Cell Division ; genetics ; Cell Line, Tumor ; Female ; Flow Cytometry ; Humans ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; genetics ; Transfection ; Transplantation, Heterologous

OBJECTIVETo analyse the causes and the management of massive hemorrhage in hepatectomy.

METHODSWith over 1 000 ml of bleeding, 4 368 patients with hepatectomy between 1955 and 2000 were analysed retrospectively.

RESULTSAmong 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.

CONCLUSIONSLight 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.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blood Loss, Surgical ; Child ; Child, Preschool ; Female ; Hemostasis, Surgical ; Hepatectomy ; adverse effects ; Humans ; Male ; Middle Aged