Objective To establish an orthotopic implantation model of human glioma in nude mice and investigate its biological features. Methods The human CHG 5 glioma cells were inoculated into brains of nude mice. The animals were sacrificed at day 8, 14, 19, 24 and 30 after inoculation. The tumors were examined with light microscope, electron microscope, karyotype analysis and immunohistochemical stain for glial fibrillary acidic protein. Results Gliomas were formed in 100% in nude mice, and the growth of tumors was stable. The tumors showed the morphological features of human glioma with the immunophenotype. Conclusion The glioma model in nude mice is a reliable animal model for the study of the tumorigenesis and biological characteristics and therapy. The 14th day after inoculation might be suitable for experimental study.

Objective To investigate the activity and mRNA expression of DNA methyltransferase(DNMT) in gliomas and the effects of chemotherapy and radiotherapy on them. Methods A total of 30 cases of glioma were divided in four groups: operation group, treatment groups A(operation+chemotherapy), B(operation+radiotherapy) and C(operation+radiotherapy +chemotherapy). Group A was treated with VM 26 and Semustine MeCCNU, B with continuous external radiation with X ray accelerated by 10MV accelerator. Fresh tissues removed surgically were used for the detection of the DNMT activity of tumors by 3H labeled tracing microassay and DNMT mRNA expression by RT PCR. Results The 2 year survival rate of the patients treated by chemotherapy or /and radiotherapy increased significantly, but the activity and the mRNA expression of DNMT decreased significantly. Conclusion DNMT is involved in the genesis and development of gliomas. The curative effect of chemotherapy and radiotherapy may be related with DNMT activity and expression. Detection of DNMT activity and DNMT mRNA may play an important role in the prognosis of gliomas and choice of treatment regimens.

Objective　To study the expressions of oncogenes c-Ha-ras, c-ki-ras, pan-ras and c-myc and point mutation of c-Ha-ras1 during hepatocarcinogenesis in rats. Methods　Immunohistochemistry, in situ hybridization and microdissection of tissue (MDT)-PCR-SSCP were used to detect the oncogene expressions and point mutation of c-Ha-ras1 in both Solt-Farber model and DEN-induced liver cancer model. Results　The overexpression of c-Ha-ras was closely associated with the formation and proliferation of the precancerous basophilic hepatocyte foci, while that of c-myc with the growth of the oval cell foci. The abnormalities of IGF-Ⅱ played an important role in the evolution of precancerous foci/nodules towards hepatocellular carcinoma (HCC). The overexpression of fms was only associated with HCC of some rats. Conclusion　Hepatocarcinogenesis in rats was related with the overexpression of c-Ha-ras, c-myc, IGF-Ⅱand fms and the point mutation of c-Ha-ras1, and overexpression of these oncogenes was associated with morphological evolution.

Objective　To explore the relationship between morphologic evolution and proliferative activity during hepatocarcinogenesis in rats. Methods　Imaging analysis technique was used to detect the morphologic parameters of cells in hepatic lesions in both Solt-Farber model and diethylnitrosamine (DEN)-induced liver cancer model. Immunohistochemistry was employed to detect proliferating cell nuclear antigen (PCNA) and bromodeoxyuridine (BrdU). Results　The oval cells were identified as irregular small proliferating cells in size of one-eighth of and with a nucleus/cytoplasm ratio of 6 times of the normal hepatocyte by image analysis. The morphometric parameters of basophil hepatocyte in precancerous foci and nodule were similar to those of the liver cancer cell. PCNA and BrdU positive cells were mainly localized within the proliferative foci and hepatocellular carcinoma (HCC) tissues. There was a better consistency between the development of hepatic lesions and cellular proliferative activity. Conclusion　The morphologic evolution is closely related to proliferative activity during hepatocarcinogenesis in rats.

Objective　To investigate the expression of glial fibrillary acidic protein (GFAP) gene and its significance in the process of glioma cell differentiation induced by nordihydroguaiaretic acid (NDGA). Methods　Immunohistochemistry (IHC) and in situ hybridization were used to detect the expression changes of GFAP protein and GFAP mRNA qualitatively and quantitatively. Results　The expression levels of GFAP protein and GFAP mRNA in NDGA treatment group were significantly increased compared with the control group (P<0.01). Conclusion　NDGA could induce GFAP gene in malignant glioma cells and the up-regulation of this gene expression might be one of the mechanisms by which NDGA induces glioma differentiation.

Thirty-six cases of astrocytoma (10 cases each of gradeⅠ and grade Ⅱ,and eight cases each of grade Ⅲ and grade Ⅳ) with definite followup data were studied with microspec-trophotometry on Feulgen stained slides to determine the nuclear area and the DNA content.10 specimens of normal brain tissue or brain tissue with gliosis were observed with the same technique.It was found that there was no significant difference of the nuclear size between the cells of grade Ⅰ astrocytoma and those with gliosis,but the DNA content was significantly higher in the former than in the latter.And there was also defference of the histograms between the 2.The higher the grading of astrocytoma,the larger the nuclear area,and the higher the DNA content.The peak values of the histograms were shifted rightward with a scattering of the values.In addition,along with the increase of DNA index,there was a decline of the survival curve of the patients with an obvious shortening of the survival time.The findings suggest that nuclear DNA quantitation can serve as an auxiliary tool to differentiate grade.I astrocytoma from gliosis,to grade astrocytomas and to predict the prognosis.

Objective To explore the relationship between the expression of pS2 protein and the clinicopathological parameters in breast cancer and to evaluate the value of pS2 as a prognostic factor for breast carcinoma and a predictive factor for response to endocrine therapy. Methods Expression levels of pS2 protein, estrogen receptor (ER) and progestogen receptor (PR) in tissues from 75 cases of breast cancer were detected by immunohistochemical assay. The relationship of pS2 protein expression with patient age, menopausal status, tumor size, metastasis to the axillary lymph nodes, pathological types, ER and PR was analyzed. Results pS2 protein, expressed in 33.3% of breast carcinomas, was correlated with patient age and pathological types, but was not correlated with tumor size. The positive rate of pS2 protein before menopause was higher than that after menopause, but no significant difference was found. Similarly, higher positive rate of pS2 protein was found in patients with metastasis to axillary lymph nodes than those without, but no significant difference was found. pS2 expression was correlated with ER and PR, but they were not perfectly consistent. Conclusion In breast cancer, pS2 protein expression, associated with some clinical factors and pathological features, is a good prognostic factor. pS2 expression may be a reasonable index for endocrinotherapy of breast carcinoma, but the detection of pS2 protein can not be employed in place of the detection of ER and PR.

Objective To investigate the effects of Nordy on the proliferation,cell cycle,and the mRNA and protein expressions of Aurora-A in human ovarian cancer cell lines 3AO and SKOV3. Methods After being treated with Nordy at the doses of 25,50 or 100 ?mol/L,the proliferation of 3AO and SKOV3 cells were tested with MTT assay; The expression of Aurora-A was detected by RT-PCR and Western blotting; The effect on cell cycle were analyzed by flow cytometry ( FCM) . Results Treated with Nordy,the mRNA and protein level of Aurora-A gene were significantly reduced ( P

Objective To observe the effects and significance of Nordy on the expression of vascular endothelial growth factor (VEGF) and inducible nitric oxide synthase (iNOS) in the retinas of diabetic rats. Methods Diabetic rats by streptozotocin were randomly divided into diabetes group (5 rats?2) and Nordy treatment group (5 rats?2). Another 10 normal rats were recruited as normal control group. The Nordy treatment group was injected 0.5% Nordy (27 mg/kg) while diabetes groups and normal control group were injected saline solution into peritoneal cavity once every other day. One month or 3 months later, 5 rats in each group were killed and the expression of VEGF and iNOS in the retinas were detected by immunohisochemistry. The average positive areas were measured and analyzed by computer aided video system. Results The expression of VEGF and iNOS in control group were extremely low. In 1 month, the expression of VEGF and iNOS in diabetic group increased and the average positive areas of VEGF and iNOS were significantly more than that of Nordy treatment group (P

Objective　To investigate the effects of TNP-470 on the growth of a human malignant glioma cell line SHG-44 in vivo and in vitro. Methods　The colorimetric MTT assay, soft agar culture, flow cytometry，light and electron microscopy were used to determine the proliferation, the cloning efficiency, cell cycle and the morphological changes of SHG-44 cells as well as the growth of its xenografted tumor. Results　TNP-470 (20～2 000 ng/ml) significantly inhibited the proliferation of SHG-44 cells in vitro (the 50% inhibitory concentration was 200 ng/ml). Cloning efficiency reduced obviously. The number of cells in G0/G1 phase increased, while that in S, G2/M phases decreased significantly. Weight and volume of xenografted tumors treated with TNP-470 (30 mg/kg, injected subcutaneously every other day) reduced notably. Furthermore, there were necrotic area and apoptosis in the tumor. No severe side effect of TNP-470 was found in this study. Conclusion　The inhibitory effect of TNP-470 on the growth of SHG-44 cells correlates with its functions of regulating cell cycle and inducing apoptosis, which suggests that the angiogenesis inhibitor TNP-470 has strong inhibitory effect on human malignant gliomas.