| According to modem scientific research, it is the basic premise of the growth of the endothelial cell (CE) in physiological and pathogen-ic antiogenesis of malignant tumor. Antiogenesis plays an important role in the growth and metastasis procedure of entity tumor. Among the factors promoting CE growth, platelet - derived endothelial cell growth factor (PD -ECGF) was found lately, and it is the new mem-ber in the field of antiogenesis. PD - ECGF/TP has the function of ab-sorbing endothelialjcell and antiogenesis, and many scholars think that PD - ECGF/TP is related with micro - vascular density ( MVD ) and plays an important role in the growth and metastasis of tumor by histol-ogy research of tumor. Transitional cell carcinoma of bladder is the most frequent malignant tumor, and its incidence rate appears increas-ing tendency. The pathogenesis, clinical biological feature and effec-tive therapy of the carcinoma have turned into the most important top-ics recently. In the paper, we evaluated the expression of PD -ECGF/TP in primary transitional cell carcinoma of bladder with immu-nohistochemical method, and revealed the relationship between the its expression and other clinical pathogenic parameters including patho-logical grade, clinical phase, MVD and the expression of p27.Experimental materials1. Selection of the specimensThe specimens were from the 72 ones embedded by paraffin of primary transitional cell carcinoma of bladder in the urology depart-ment of No. 1 Affiliated Hospital of China Medical University from Jan 2001 to Apr 2002. All the specimens were identified by pathological doctors. The patients includes 61 males and 11 females ( aged from 30 to 84, mean 61. 6). According to WHO criteria of carcinoma cell grade, there were 15 cases in Gl phase, 30 cases in G2, and 27 cases in G3. According to UICC - TNM criteria of clinical phase, there were 28 cases with superficial carcinoma of bladder ( Ta, T,) , and 44 cases with infiltrated carcinoma of bladder (T2 - T4). The five normal blad-der mucous membranes were from the patients performed with open op-eration of benign prostate hyperplasia. All the specimens were routine serial section, whose thickness was 5$m.2. Main agentsThe main agents supplied by Fuzhou Maxim Biotech Inc. inclu-ding : rat ?anti - PD - ECGF monoclonal antibody, rat - anti - PD -ECGF M factor related monoclonal antibody, rat - anti - p27 monoclonal antibody, S - P agent box, DAB agent.Experimental method1. HE staining; All the sections were performed HE staining.2. Tissue staining with immunohistochemical S -P method; The sections embedded by paraffin were performed routine de -wax with dimethylbenzene, watered by alcohol, cleaned by PBS one time, and stored in 3% hydrogen peroxide (A fluid) thermo -box for 30 min. The specimens were cleaned for 3 x 3 min, and applied hyper - pressure antigen repairing. Each section were cooled in tap - water for 20 - 30 min, and added with one dribbling animal non - immuno-logical serum ( B fluid) and incubated in thermo - box of 37Tl for 30 min. Threw B fluid away, and added one dribbling of first antibody, and stayed for one night. Watered each section with PBS for 3x5 min, added one dribbling of second antibody (C fluid) and incubated in thermo - box of 37?C for 30 min. Watered each section with PBS for 3 x5 min, added one dribbling of second antibody (D fluid) and incubated in thermo - box of 37 Tl for 30 min. Watered the section for 3x5 min with PBS fluid, showed DAB color, watered with tap - wa-ter, re - stained with hematoxylin, dried with gradient alcohol, cleared with Dimethylbenzene and mounted with neutral gum.3. Control group(1) Negative control: normal mucous membrane of urinary blad-der and PBS substituted tumor tissue specimen of first antibody.(2) Positive control; positive control photograph were bought.4. Result determinant(1) Determinant of PD - ECGF/TP expression: Determine ac-cording to staining intensity of cells and accumulating of the total area of staining... |
