| Glioma is the most common tumor of the brain. The special location and invasive growth of astrocytic tumors result in its difficulties in treatment. Since glioma is usually active in angiogenesis which is related to the tumor growth and progression, it has been often used for tumor angiogenesis. Therefore, it is very necessary to further study glioma angiogenesis and its mechanism for the anti-angiogenesis strategy. High-density tissue microcrray, or tissue chip, is a newly-established technology during the past six years. Tissue chip make it possible to analyze up to hundreds or thousands of specimens on a same vector at the same time. Tissue microarray can be used to study the expression of gene and protein, and it has an important role in studying the expression of disease-related gene and protein, molecular diagnosis, evaluating the effect of treatment and screening of new drugs.In this study, tissue microarray containing different human astrocytoma microvessels was constructed and used to evaluate of astrocytoma angiogenesis. In addition, computer-aided three-dimensional reconstruction of malignant glioma microvessels was established. The main results and conclusions are as follows:1. A practical method of tissue microarray construction was established with which a tissue microarray of astrocytoma microvessels (AstMV) was successfully constructed.(1) The first step of tissue microarray construction was the selection of the typical areas with special vascular structure, which was followed by orientation of donor blocks, preparation of acceptor block and use of tissue microarray apparatus. AstMV tissue microarray were constructed . For the necessity of the making of tissue microarray, a system for location was made, which enhanced the efficiency and standardization of making tissue microarray.(2) Experiences in tissue microarray construction was obtained, such as special treatment of "receptor" blocks, gentle operation, controlling the depth of the needle and the treatment of the tissue microarray block.2. The AstMV tissue microarray was applied to analyze in vivo astrocytoma angiogenesis The tissue microarrays constructed in our study were used to detect the expression of CD34, â…§-R Ag, CD105, VEGF and GFAP. The results showed that there was difference in microvessel number between astrocytic tumors and between endothelial markers. In the astrocytoma grade â… , no difference in statistics could be found in the detection for CD34, â…§-R Ag and CD105. In the gradeâ…¡,â…¢ and â…£ astrocytomas, labeled microvessels by CD34 and â…§-R Ag antibodies were more than those detected by CD105 antibody (P<0.01). There were differences in labeled microvessels by CD34 and â…§-R Ag antibodies between grades â…¡,â…¢ and â…£ tumors (P<0.01). However, the microvessels stained with CD105 antibody had no difference (P>0.05) between the tumor grades. These results suggested that the three marker for endothelial cells should be selected for use in angiogenesis according to different purposes. Immunohistochemical staining for VEGF showed that its expression became increased along with the increasing of tumor grades, while GFAP expression showed that its expression became decreased along with the increasing of tumor grades.All the above results were accorded with those results on the routine slides. It suggested that AstMV tissue microarray could actually represent the original in vivo tumor blocks and have its advantages in analysis of angiogenesis because of its easy quanlity control.3. 3D reconstruction of malignant glioma microvessels was obtained on the serial tissue sections with a computer-aided system.(1) The basic method of 3D reconstruction of malignant glioma microvessels using serial sections was established. The serial sections of one case of human astrocytoma were used to reconstruct its 3D microvessel figure. The main protocols of 3D reconstruction of tumor microvessels included serial sectioning, registration, labeling of endothelial cells with FVIII Rag, collection and proceeding of figures and 3D...
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