To investigate the effects of gambognic acid (GA) on TRAIL-induced apoptosis of cancer cells, human colon HT-29 cancer cells were treated with GA to promote apoptosis. Inhibition of the cell proliferation was measured with MTT assay and cell apoptosis was detected with formation of DNA ladders in agarose gel electrophoresis, and activation of caspase activity. The content of cytosolic reactive oxygen species (ROS) was measured with flow cytometry. The activities of Caspase-3, -8, -9 were detected using spectrophotometric assay. The levels of c-FLIP, CHOP, DR4 and DR5 in cells were tested by Western blot. Combination of GA (1 µg · mL(-1)) and TRAIL (40 ng · mL(-1)) significantly reduced proliferation and increased apoptosis of HT-29 cells over those induced by each agent alone. Percentage of apoptotic cells was increased to 45.5%. GA markedly enhanced the intracellular ROS generation. Expression of CHOP, DR4 and DR5 was up-regulated to 7.38, 5.41, and 4.85 times of the control group, respectively. GA promoted activation of Caspase-3, -8, and -9 by TRAIL (P<0.05). Furthermore, the expression of anti-apoptotic protein c-FLIP was down-regulated to 0.22 ± 0.08 times of the control group. In conclusion, GA sensitizes HT-29 cells to TRAIL-induced apoptosis by promoting ROS-activated ERS pathways, up-regulating of DR4 and DR5, and inhibiting c-FLIP expression.
Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Caspases ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; metabolism ; Down-Regulation ; HT29 Cells ; Humans ; Reactive Oxygen Species ; metabolism ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; Up-Regulation ; Xanthones ; pharmacology

Nanoemulsion-encapsulated lzp3 DNA vaccine (pCDNA3-Aat-COMP-lzp3-C3d3, lzp3) was prepared, and its quality was evaluated. The interfacial emulsification method was employed to make the nanoemulsion-encapsulated plasmid, and its sizes and distribution were detected by electron microscope. Anion exchange chromatography was used to separate the nanoemulsion from the plasmid based on the charge characteristics of plasmid. The determination method for encapsulation efficiency of the plasmid nanoemulsion was established by anion exchange chromatography. The results indicated that the average size of the plasmid nanoemulsion is (23 +/- 10) nm, and the encapsulation efficiency is 80.5%. The separation and determination conditions of nanoemulsion-encapsulated plasmid was as follows: eluent buffer was 0.05 mol/L Tris-HCl solution at a flow rate of 0.7 mL/min, while detection wavelength for plasmid was 260 nm, the temperature of column was maintained at 30 degrees C, and injection volume was 2mL. On this condition, nanoemulsion and plasmid can be separated on the column of anion exchange chromatography significantly. This method is simple, sensitive and reproducible with respect to good linearity in the range of 0.05-0.80 mg/mL (r = 0.9983), which can be applied to determine the entrapment efficiency of nanoemulsion-encapsulated plasmid.
Animals ; Chromatography, Ion Exchange ; methods ; Drug Carriers ; chemical synthesis ; chemistry ; Emulsions ; Nanotechnology ; Plasmids ; chemistry ; Rats ; Vaccines, DNA ; chemistry ; Zona Pellucida ; immunology

Objective To explore the establishment of a rat model of acute radiation-induced liver injury and sig-nificance of the dynamic changes of TGF-β1 expression.Methods Forty healthy 6-week old male SD rats were randomly divided into model group (n=30) and control group (n=10).The right liver of rats in the model group was given a single dose of 25 Gy 6 MV X-ray irradiation.Histopathological examination using HE staining and transmission electron microsco-py were conducted to observe the liver pathological changes in rats at 3, 5, and 10 days after irradiation, serum TGF-β1 was detected, and relevant indicators of liver function ( ALT, AST, ALP) were determined.Statistical analysis was per-formed using SPSS 17.0 software.Results At 3, 5 and 10 days after irradiation, early pathological changes in the liver cells were observed by electron microscopy, the expression of TGF-β1 was gradually increased with the time prolongation, and significant differences were found between the model group and the control group at different time points (P<0.05). The light microscopic observation of liver tissues did not show significant differences between the control group and model group.The liver ALT, AST, ALP at different time points did not show significant differences between the two groups ( P>0.05).Conclusion Electron microscopy can be used to evaluate the early changes of radiation-induced liver injury, pri-or to the alterations visible by routine light microscopy.TGF-β1 can be used to predict the degree of radiation-induced liver injury, and may be used as a sensitive serum cytokine in predicting the degree of radiation-induced acute liver injury.

OBJECTIVETo explore the pattern of lymph node metastasis and its influence on the prognosis of early gastric cancer(EGC).

METHODSThe pattern of lymph node metastasis and the 3-,5-year survival rates in 157 EGC patients undergone surgery from October 1995 to October 2005 were analyzed retrospectively. The SPSS 11.5 statistics software was used to perform univariate and multivariate analysis.

RESULTSTwenty-two cases had lymph node metastasis among 157 EGC patients(14%). Two mucous cancers(2.4%) and 20 submucosal tumors(27.0%) had lymph node metastases (P<0.01). Lymph node metastasis was not seen in minute gastric cancer(diameter < or =0.5 cm). Lymph node metastasis rates were 6.4% in the cancers with diameter 1.1-2.0 cm and 21.5% in the cancers with the diameter >2.0 cm(P<0.01). Besides, lymph node metastasis rate of well-differentiated EGC was 0, of moderate differentiated EGC 11.1%, and poor-differentiated EGC 0.9%(P<0.01). Of 9 cases with vascular cancer embolus, 4 had lymph node metastases. Logistic regression analysis showed that tumor size, vascular cancer embolus, histopathological type and depth of invasion were independent factors of lymph node metastasis in EGC. The 3- and 5-year survival rates of EGC patients with lymph node metastasis were 81.6 % and 79.5% respectively, which were much lower than those without lymph node metastasis(95.7% and 93.2%, P<0.01).

CONCLUSIONSLymph node metastasis in EGC is mainly correlated with depth of infiltration, tumor size, vascular cancer embolus and differentiation. For EGC treatment, choice should be made reasonably based on the risk of lymph node metastasis.

Adult ; Aged ; Female ; Humans ; Logistic Models ; Lymphatic Metastasis ; diagnosis ; pathology ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Prognosis ; Retrospective Studies ; Stomach Neoplasms ; diagnosis ; pathology ; Survival Rate

Anticarcinogenic Agents ; administration & dosage ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Non-Small-Cell Lung ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Isothiocyanates ; administration & dosage ; isolation & purification ; pharmacology ; Lung Neoplasms ; pathology ; Plants, Medicinal ; chemistry ; Protein Binding ; Raphanus ; chemistry ; Tubulin ; chemistry ; metabolism

OBJECTIVETo investigate the correlation of CD4CD29regulatory T cells (Treg) with tumor recurrence and survival time in patients with non-small cell lung cancer (NSCLC).

METHODSFifty-nine patients with NSCLC treated with radical surgery were followed up for 5 years. Blood Treg cells were examined during the follow-up using flow cytometry (FCM). The sensitivity and specificity of Treg cells to predict recurrence of NSCLC were analyzed using receiver-operating characteristic (ROC) curve and compared with those of carcinoembryonic antigen (CEA) and cytokeratin21-1 (Cyfra21-1). The influences of gender, age, occupation and radiotherapy on survival time of the patients were analyzed with Kaplan-Meier method.

RESULTSAmong the 59 patients, the shortest survival time was 23 months while the longest time was over 67 months. Nineteen patients had NSCLC recurrence, and 17 (28.81%) of them died of metastasis during the follow-up. The frequencies of blood Treg cells in patients who did not receive radiotherapy and in patients with tumor recurrence were significantly higher than those in patients receiving radiotherapy and in patients free of recurrence (P=0.000). ROC curves showed that the area under curve (AUC) lowered in the order of Treg cells, Cyfra21-1, CEA (P=0.002, 0.006 and 0.013, respectively) with 95% confidence interval (CI) of 0.649-0.981, 0.621-0.936 and 0.584-0.944, respectively. At the cut-off value of 7.53%, the sensitivity and specificity of Treg cells to predict NSCLC recurrence was 91.42% and 87.59%, respectively. The five-year survival rate of the 59 patients was 71.18% (42/59), and Kaplan-Meier analysis revealed a longer survival time in female patients (P=0.038), in patients below 50 years of age (P=0.013), in patients not engaging in mental work (P=0.029), and in patients receiving radiotherapy (P=0.003).

CONCLUSIONTreg cells has a better efficiency than Cyfra21-1 and CEA to predict tumor recurrence in patients with NSCLC following radical surgery. The male gender, an age beyond 50 years, an occupation of mental work, and failure to receive radiotherapy are all risk factors for recurrence of NSCLC.

OBJECTIVETo investigate the influence of ICAM-1 469K/E gene polymorphisms on the risk of atrophic gastritis and dysplasia.

METHODSThe ICAM-1 469K/E gene polymorphisms in a total of 372 subjects were detected by polymerase chain reaction-direct sequencing. All of the subjects were from Linqu County, a high risk area of gastric cancer in Shandong Province of northern China. All cases were initially diagnosed as normal or superficial gastritis at the beginning of this study. After a 5-year follow-up, the cases were subdivided into no progression group (no histological progression, n=137), progression group I (progressed to severe chronic atrophic gastritis, n=194) and progression group II (progressed to low-grade dysplasia, n=41).

RESULTSIn all 372 subjects, the frequencies of KK, KE or EE genotype of ICAM-1 K469E were 50.5%, 39.2% and 10.2%, respectively. No significant differences were observed in the ICAM-1 469K/E genotype frequencies between the progression group I and no progression group (P>0.05). The frequencies of KK genotype (68.3%) were significantly higher in the progression group II than in the no progression group (49.6%, P=0.035), and also than in the progression group I (47.4%, P=0.015). An increased risk of the progressing to dysplasia from normal or superficial gastritis was found in the individuals with ICAM-1 469KK genotype [odds ratio (OR)=2.21, 95%CI, 1.10-4.42].

CONCLUSIONICAM-1 469K/E gene polymorphisms are significantly associated with the risk of gastric low-grade dysplasia, but not related with severe chronic atrophic gastritis in a population with high risk of gastric cancer in Linqu County, Shandong Province, China.

Adult ; Female ; Follow-Up Studies ; Gastritis ; genetics ; pathology ; Gastritis, Atrophic ; genetics ; pathology ; Genotype ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Male ; Middle Aged ; Odds Ratio ; Polymorphism, Genetic ; Precancerous Conditions ; genetics ; pathology ; Risk ; Stomach Neoplasms ; genetics ; pathology

OBJECTIVETo examine the utility of the Ligasure vessel-sealing system in D(2) lymphadenectomy of gastric cancer, and compare with conventional hand tie method.

METHODSOne hundred and twenty-four consecutive patients undergone D(2) lymphadenectomy of gastric cancer from Jan. to Oct. 2005 were enrolled in this study. Operations of 62 patients were performed with Ligasure, whereas the other 62 patients with hand tie method. Operative duration, volume of intraoperative hemorrhage and postoperative course were analyzed.

RESULTSThere was significant difference in operative duration (187.3 min vs 210.5 min, P< 0.05) and no significant differences in volume of intraoperative hemorrhage, postoperative course, or duration of postoperative drainage between Ligasure group and conventional method group. No significant difference was observed in the frequency of postoperative complications between the two groups.

CONCLUSIOND(2) lymphadenectomy of gastric cancer using Ligasure instead of conventional hand tie method appears to be technically feasible and easy.

Female ; Hemostasis, Surgical ; methods ; Humans ; Ligation ; Lymph Node Excision ; Male ; Middle Aged ; Neoplasm Staging ; Stomach Neoplasms ; pathology ; surgery

OBJECTIVETo suggest a chemical surface treatment for titanium and to initiate the formation of hydroxycarbonated apatite (HCA) on titanium surface during in vitro bioactivity tests in simulated body fluid (SBF).

METHODSTo improve the bone-bonding ability of Ti implants, commercially pure titanium (cpTi) by a simple chemical pre-treatment in orthophosphoric acid (H(3)PO(4)) with different density was activated, and then the phosphorylation specimens were soaked in SBF to investigate the function of biomineralization.

RESULTSThe scanning electron microscope (SEM) photographs showed that the surfaces of the pre-treated samples were characterized by a complex construction, which consisted of a mesh-like morphology matrix (a micro-roughened surface) and an uniform surface with different morphous of titanium dihydrogen orthophosphate [Ti(H(2)PO(4))(3)] crystal. After 14 days in SBF a homogeneous biomimetic apatite layer precipitated.

CONCLUSIONSThese data suggest that the treatment of titanium by acid etching in orthophosphoric acid is a suitable method to provide the titanium implant with bone-bonding ability.

Acid Etching, Dental ; methods ; Biomimetics ; Body Fluids ; Coated Materials, Biocompatible ; Dental Bonding ; Dental Implants ; Microscopy, Electron, Scanning ; Phosphoric Acids ; chemistry ; Phosphorylation ; Surface Properties ; Titanium ; chemistry