Objective To determine the inhibitory effect of APE1 siRNA expression vector pSilence APE1, and the possible synergetic role of pSilence APE1 and endostatin on endothelial cell migration induced by osteosarcoma cell 9901 and HOS . Methods The osteosarcoma cells and endothelial cells were co-cultured with transwell model, and the cell number through the inner membrane was counted to evaluate the inhibitory effect of endothelial cell migration by pSilence APE1 and its combination with endostatin. Results Both low dose (350 ng/ml) and high dose (700 ng/ml) of endostatin showed significant inhibition to endothelial cell migration induced by osteosarcoma cell, while the high dose showed much stronger inhibition than the low dose (P

Objective To detect the expression of apurinic/apyrimidinic endonuclease 1 (APEI) and explore its correlation with the expression of mutant p53 in hepatocellular carcinoma (HCC). Methods The expression of APE1 and mutant p53 was detected by SP immunohistochemical method in 10 specimens of normal liver tissue, 40 specimens of liver cirrhosis tissue and 103 specimens of HCC tissue which were collected at the Department of Pathology of Daping Hospital from 1991 to 2004. All data were analyzed by chi-square test, correla-tion analysis and K Independent-Samples Tests. Results The expression rate of APE1 in HCC was 100.0%, which was significantly higher than that in normal liver tissue (40.0%) and liver cirrhosis tissue (82.5%) (χ~2= 47.852, P < 0.01). The expression of APE1 was only detected in the nucleus in normal liver tissue. Ectopic expression of APE1 in cytoplasm was detected in liver cirrhosis tissue and HCC tissue, with the rate of 20.0% and 53.4%, respectively (χ~2=20.757, P <0.01). There was statistical difference in clinical staging and pathological grading of HCC with different combinations of APE1 expression (intranuclear or ectopic expression) and mutant p53 expression (positive or negative expression) (χ~2=12.910, 14.481, P < 0.01), and HCC with ectopic expression of APE1 and positive expression of p53 had high malignant degree. Conclusion Overexpression and ectopic expression of APE1 in cytoplasm may play important roles in the genesis and progression of HCC, and the ectopic expression of APE1 and p53 mutation may have synergistic effect.

Objective To investigate the value of combined detection of multi-tumor markers in the diagnosis of primary hepatocellular carcinoma (HCC) and to establish the discriminant equation. Methods Using a protein chip, 12 tumor markers in the serum from 98 patients with HCC and 67 patients with benign liver diseasewho had been admitted to Daping Hospital from November 2003 to April 2006, and 46 healthy individuals during he same period were analyzed. A discriminant equation was established to discriminate primary HCC from benign liver diseases. All the data were processed by variance analysis and chi-square test. Results The positive rates of the tumor markers were 89% (87/98) in patients with primary HCC, 19% (13/67) in patients with benign liver disease and 4% (2/46) in healthy individuals. There was statistical difference in the serum level of alpha-fetoprotein (AFP), eareinoembryonic antigen (CEA), ferritin (FER), CA19-9 and CA125 among the 3 groups (F =59.530, 40.472, 31.708, 75. 897, 153.066, P <0.05). Combined detection of AFP, CEA, FER, CA19-9 and CA125 improved the diagnostic accordance rate to 89%, which was significandy higher than the diagnostic accordance rate (64%) when only AFP was detected (X2 = 16.362, P <0.05). The accuracy of the discriminant equation was 90%. Conclusions Combined detection of multi-tumor markers is superior to AFP detection. Combined detection of multi-tumor markers can be used in screening of the HCC patients in HCC high risk population and in the early diagnosis of primary HCC.

Objective To study the expressions and correlation of cyclooxygenase-2 (COX-2) and epidermal growth factor receptor (EGFR) in human transitional cell carcinoma of bladder (TCC) tissues and explore the correlation of their expressions in the incidence and development of TCC. MethodsThe expressions of COX-2 and EGFR were detected by immunohistochemical technique in 48 cases of TCC and in 10 cases of normal bladder mucosa tissues. ResultsThe expressions of COX-2 and EGFR in TCC was significantly higher than those in normal bladder mucosa tissues (P

Objective To study the effect of caffeic acid phenethyl ester (CAPE) on the expression of ?-catenin in the cultured colorectal cancer cell lines. Methods HCT116 and W480 cells were treated with CAPE at serial concentrations of 2.5, 5, and 10 mg/L. ?-catenin protein expression was assayed by Western blot analysis. ?-catenin localization was detected by indirect immunofluorescence. Results CAPE treatment was associated with decreased total ?-catenin protein expression. The expression of ?-catenin at the cell nucleus and cytoplasm was downregulated, but at the cell-cell linked site the ?-catenin protein expression was upregulated. Conclusion CAPE can downregulate the expression of ?-catenin and inhibit the translocation of ?-catenin to nucleus, which may play an important role in the anticancer activity of CAPE.

Objective To construct DNA damage and repair gene apurinic/apyrimidinic endonuclease (APE1) siRNA expression vector pSilence APE1 and investigate its inhibitory effect on the expression of APE1 in osteosarcoma cell HOS and 9901 in vitro. Methods An expression plasmid of a short hairpin RNA target APE1, pSilence APE1 was constructed and transfected to 9901 and HOS cell by lipofectamine. The expression of APE1 protein in HOS and 9901 osteosarcoma cells posttransfected with pSilence APE1 was detected by immunohistochemistry. Meanwhile, the dose-effect and time-effect relationship of APE1 gene silence induced by pSilence APE1 were measured using Western blot analysis. Results After evaluation and sequencing, the APE1 siRNA expression vector pSilence APE1 was constructed successfully. The result of Western blotting and immunohistochemistry showed that APE1 protein in osteosarcoma cells could be knocked down specifically by pSilence APE1, and the inhibition rate of APE1 expression was 72%-95%. The best inhibition of expression of APE1 gene was 3.0 ?g and at the 72 h using pSilence APE1. Conclusion APE1 siRNA expression vector pSilence APE1 is successfully constructed that can significantly knock down APE1 gene expression in osteosarcoma cells.

BACKGROUND: Recently, some people believed that the mechanisms of primary hepatic carcinoma might be caused by poor differentiation or disdifferentiation of hepatic stem cells. Studies on hepatic stem cells are in the early stage at present, and the theory of "stem cell origins" of human primary hepatic carcinoma deserves further verification. OBJECTIVE: To investigate the activation, distribution, origin and immunological expression characteristics of hepatic stem cells in different histopathologic types of primary hepatic carcinoma. DESIGN: Observational comparative study. SETTING: Tumor Center, Research Institute of Surgery, Daping Hospital, Third Military Medical University of Chinese PLA. PARTICIPANTS: Experiments were performed at the Laboratory of Tumor Center, Research Institute of Surgery, Daping Hospital, Third Military Medical University of Chinese PLA from September 2003 to July 2004. We took 94 cases of hepatic cellular cancer, 12 cases of intrahepatic cholangiocellular carcinoma and 10 cases of mixed hepatocarcinoma paraffin-embedded tissue blocks as research objects, with 5 cases of liver cirrhosis and 4 cases of normal liver as experimental control. These materials were collected from the archive of the Department of Pathology of Daping Hospital. Primary hepatic carcinoma tissues and corresponding adjacent liver tissues were obtained from patients who had undergone surgery for the removal of their tumors. All the patients were not treated by chemotherapy or radiotherapy before the operation. They had signed the informed consent. Main Antibodies were bought from Santa Cruz Company.METHODS: The histological and immunohistochemical characteristics were examined by haematoxylin and eosin staining and immunohistochemistry (SP method), including mouse antihuman cytokeratin 19 monoclonal antibody, mouse antihuman cytokeratin 7 monoclonal antibody, mouse antihuman cytokeratin 8&&18 monoclonal antibody, mouse antihuman c-kit monoclonal antibody, mouse antihuman Thy-1 monoclonal antibody, mouse antihuman alpha fetoprotein monoclonal antibody. MAIN OUTCOME MEASURES: Expression of immunological markers of hepatic stem cells in different histopathologic types. RESULTS: Immunological markers of hepatic stem cells expressed variously in different histopathologic types of primary hepatic carcinoma. Hepatic stem cells differentiated into hepatoma carcinoma cells in all the types. The highest expression rate of hepatic stem cell immunophenotype was found in the mixed hepatocarcinoma (P < 0.05). Immunophenotypes of hepatic stem cells were negative in normal group and cirrhosis group. CONCLUSION: Hepatic stem cells of varied differentiations and origins existed in different histopathologic types of primary hepatic carcinoma.

Objective:To assess proton biological dose with using two kinds of relative biological effect models and to compare them with traditional clinical proton biological dose ( Dose1.1). Methods:Based on Particle simulation tools(TOPAS), physical dose, LET d and LET t were calculated in water phantom and two anthropomorphic phantoms (brain and prostate tumors) respectively. Then DoseLET d and DoseLET t were calculated according to different relative biological effect models, an RBE was 1.1 in traditional clinical proton biological dose calculation. Three kinds of biological doses were compared in the water phantom. To quantify the differences between three method in anthropomorphic phantoms, three points ( D1, D2, D3) were selected according to the physical dose to compare the biological dose. Results:DoseLET d and DoseLET t in water phantom showed the same trend with water depth and both of them were higher than Dose1.1 at the end of proton beam range. The maximal difference between DoseLET d and DoseLET t in the anthropomorphic phantoms was 10.08 cGy, where the relative difference was less than 5%. When DoseLET d and DoseLET t were compared with Dose1.1, the maximal differences in brain tumor target were 71.97 cGy and 61.91 cGy respectively, where the relative differences were less than 25%. The maximal differences in prostate tumor target were 25.95 and 19.96 cGy respectively, where the relative differences were less than 12%. However, the differences outside the target were very small, where the maximal differences in brain and prostate tumors were 5.99 cGy and 9.92 cGy respectively, and the relative differences were less than 5%. Conclusions:Biological doses calculated by two method are of little difference in both water and anthropomorphic phantoms, however, large differences were observed when they were compared with the traditional clinical proton biological dose especially in the high dose area.

Rheumatoid arthritis （RA）， as an autoimmune disease， is mainly characterized by persistent synovitis. It often involves multiple joints symmetrically and can lead to joint deformity， joint function loss， and even disability in severe cases. The pathogenesis of RA is complex， and the prevention and treatment are complicated. Therefore， it is difficult to cure the disease completely. Previous studies have validated important targets and mechanisms for the prevention and treatment of RA， including the nuclear factor-κB （NF-κB） signaling pathway that controls the inflammatory process， nuclear factor E₂-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1） signaling pathway that regulates oxidative stress， inhibits inflammation， and maintains cell homeostasis， Wnt/β-catenin signaling pathway that plays a key role in cell growth， differentiation， apoptosis， and inflammatory response， anti-inflammatory， anti-oxidation， and silent information regulator 1 （SIRT1） signaling pathway that regulates synovial cells， anti-inflammatory adenylate-activated protein kinase （AMPK） signaling pathway that regulates energy metabolism， and hypoxia-inducible factor-1α （HIF-1α）/vascular endothelial growth factor （VEGF） signaling pathway related to angiogenesis in RA. At the same time， many studies have confirmed that traditional Chinese medicine prevents and treats RA by regulating the above signaling pathways and exerting their related effects， indicating the advantages of traditional Chinese medicine such as multiple regulatory pathways， long-term effects， and less adverse reactions. In this paper， by consulting many research reports， the role of the above-mentioned signaling pathways in RA was clarified， and the latest research results of traditional Chinese medicine intervention in the above-mentioned signaling pathways in the prevention and treatment of RA in recent years were summarized in detail. This paper aims to promote the in-depth study of the pathogenesis of RA and its treatment with traditional Chinese medicine， provide a scientific basis for the rational application of traditional Chinese medicine， and offer useful enlightenment for the development of new drugs and clinical practice for the treatment of RA in the future.