| Gastric cancer is the most common malignant tumor and the first frequent cause of cancer death in our country. Though the incidence rate of gastric cancer has slightly decline in past years, it is one of malignant tumor that hazard the people's health of our country at present. Although the background theorem and clinic investigation of gastric cancer has gained more achievement, advanced gastric cancer will develop metastasis early after operation, 5-year survival rate of patients with gastric cancer is still poor. An improvement in survival and prognosis of gastric cancer may be achieved by selecting colligate therapeutics reasonably. Now, the selection of chemotherapy program of gastric carcinoma is mainly depend on tumor infiltration as well as lymph node metastasis. Thymidine phosphorylase is one of the pyrimidine nucleoside phosphorylase family, it is essential for synthesis and restriction of cell growth. Dihydropyrimidine dehydrognenase make 5-fliorouracil inactivate by catalysis 5-fliorouracil to 5-FUH2 .It is make clear now, thymidine phosphorylase is identical to platelet derived endothelial cell growth factor with structure and function. TP posses highly activity in most solid tumor and relation with tumor angiogensis, faster growth and ability of infiltration, and it can prevent tumor cell apoptosis, relation with bad prognosis. In additional, expression of thymidine phosphorylase is regulated by some cell factor, drug, agent of physics and chemistry. In present study, the concentrations of thymidine phosphorylase were determined using ELISA(enzyme-link immunosorbent assay) in tumor and normal tissue of gastric cancer; Microvssel density(MVD)were observed using immunohistochemical(Envision assay) in tumor and normal tissue. The relation ship between the concentration of thymidine phosphorylase of gastric cancer and normal tissue, the concentration of thymidine phosphorylase of gastric cancer as well as pathologic parametes, and tumor thymidine phosphorylase as well as tumor microvessel density were analyzed,,Materials and methods1. PatientsrFrom May,2001 to December,2001, 45 surgical specimens were obtained from patients with gastric cancer, included 33 male and 12female,ranging in age from 36 to 78 years(mean 62). Operation manier: 23 cases for distal subtotal gastrectomy, 8 cases for proximal subtotal gastrectomy , 8 cases for total gastrectomy (include unite organs resection) .fresh tumor tissue and normal gastric mucous membrane(300mg) were removed and stored at -80 'C for IHC.2. Reagents: TP, DPD EL1SA kit was a generous gift from ShangHai Roche pharmacenticals Ltd; CD34 mouse anti-human monoclonal antibody was purchased from Zhong shang Biotech Ltd, Beijing China; Envision kit was purchased from DAKO Ltd, USA.3. Methods3.1 EL1SA methods: frozen tissue was homogenize shocked,, resuspend homogenate in lOmMol Tris buffered solution, centrifuge at lOO.OOOg for 60min at 2-8 癈, 1:2 dilutio supermatant. TP and DPD concentration were determined using ELISA based on the demands of TP and DPD kits.3.2 Envision method stained: Surgical specimens of gastric cancer and normal gastric tissue were fixed in formalin, embedded by paraffin and sliced at Sum 1 intriplicate. One slice was stained by HE and the others were stained by method based on the demands of the Envision kit.4. Result Determination4.1 ELISA assay : tumor and normal tissue OD, standard curve was constructed by, semilogarithmic regression equation based on the demands ofTP and DPD kits. Calculate TP and DPD concentration. Unit: ng/ml.4.2 MVD count method: based on the standard of Weinder: one brown endothelium cell or endothelium cluster as one blood vessel count unit. Elimination blood vessel muscular layer thicker and area of blood vessel cavity >8 cell diameter. The estimated of MVD was determined in 5 random areas of one section at 10><20, counted, calculate mean MVD. 5 Statistics analysis5.1 Wilcoxon signed ranks test was used to analyse the correlation of TP, DPD concentratio... |
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