ObjectiveTo research the conditions of Human Umbilical Vein Endothelial Cells (HUVEC) primary culture and the effect of thalidomide on HUVEC in order to provide theoretical basis for studying the development of treatment for chorodidal neovascularization (CNV). To determine if thalidomide inhibit experimental fibrovascular proliferation induced by Krypton laser in mice as the model of CNV. To evaluate the toxicity of thalidomide through the change of body weight and behavior of the mice. MethodsThere are two parts in this study which includes thalidomide is used in HUVEC and the model of CNV.1 Cell-based experiment1.1 The primary culture of HUVEC The umbilical cord was washed carefully. From one end of the cord, we filled the cord with 0.1% collagenase I. 10min later, the liquid was collected in the cord and implanted into a centrifuge tube, 1000r/mim for 7 min. Then, we transferred the cell suspension to a culture plate which was placed in an incubator at 37°C with 5% CO_2. HUVEC was identified by means of immunohistochemical staining for factor Ⅷ related antigen (FⅧ).1.2 Effect of thalidomide on HUVEC HUVECs were randomly devidedinto 3 groups which included group A, B and C. Group A: thalidomide was used with following concentration gradient: lOug/ml, 25ug/ml, 50ug/ml, and 75ug/ml. Group B: Dexamethasone (lug/ml) was added into the thalidomide in different concentration respectively. Group C: It was as the control group. We measured the absorbance ratio through MTT-test. Date were applied for statistical analysis on computer with software SPSS 11.0. 2 Animal-based experiment2.1 Effects of thalidomide on the experimental CNV of Mouse Through Krypton laser, we establish CNV model to 17 health C57BL/6J mouse. Krypton laser parameters: wavelength 647nm, power 120mW, and exposure time 0.1s, spot size 50um. Group A: 5 mice, comparison group. Group B: 6 mice, low concentrations group, abdominal cavity injection thalidomide (lOmg/kg) after laser photocoagulation. Group C: 6 mice, high concentrations group, abdominal cavity injection thalidomide (50mg/kg). All groups abdominal cavity injection drug for 10 days. Then, kill the mice with spinal cord dividing. Serial histological specimens were methodically assessed in an image pattern analysis, maximum FVP thickness for each lesion was measured. We use software SPSS 11.0 for statistical analysis.2.2 Toxicity of thalidomide to mouse Record the change of mouse's body weight before and after thalidomide injection respectively. We detect the change of pathergasiology of mouse with defense reflex, the difficulty of capture, food intake, color of hair and mental state. Each side is divided into three different levels.Results1 Cell-based experiment1.1 The primary culture of HUVEC For 6 hours later, cells can adherent and growth;For 48 hours later, most of cells was polygon, some was fusiform, entoblast was distinct;For 72 hours later, cells filled the basement of culture plate, most of cells was fusiform, entoblast was distinct. When identify the cell, we can seeentoblast was blue staining, a great quantity of yellow-green grain can be seen intracytoplasmic.1.2 Effect of thalidomide on HUVEC Group A and B can inhibit the growth of HUVEC significant (p<0.01);Group B had a strong inhibition effect to HUVEC than Group A. With the concentration increase of thalidomide, drug potency was intensive. Thalidomide potency is dose related.2 Animal-based experiment2.1 Effects of thalidomide on the experimental CNV of Mouse In treated eyes the development of CNV was significantly inhibited. Group A: lots of CNV was found in lesion, FVP was thicker than other groups. There were exudative retinal detachments in most eyes;lesion was encysted by retinal pigment epithelium (RPE) which was obtrite, the photoreceptor cell was inordinate. Group B: lesser CNV in lesion;there were exudative retinal detachments in some eyes;some anomalous vessel lumen could be seen. RPE cells were inordinate, some photoreceptor was disappearing. Group C: the manifestation was the same as Group B. The FVP thickness of Group A was thicker than Group B and C, the difference was significant, but there was no difference between Group B and C.2.2 Toxicity of thalidomide to mouse 10 days late after thalidomide injection, there was significant difference about the change of body weight of the mouse, the change of Group C was less than A and B. In pathergasiology, there were no change between Group A and B, but the mouse of Group C have had violent defense reflex, the less food intake and sunken.Conclusions1. We can get sufficient HUVEC successfully using this method.2. Thalidomide can inhibit the growth of HUVEC significantly. Drug resulted in does-dependent changes in HUVEC.3. This study provides evidence that thalidomide effectively inhibited CNVprogression in a mouse model of laser-induced CNV.4. Mouse can tolerance routine dosage of thalidomide. However, when over-dosage thalidomide was used, inhibition of CNV did not be improved obviously, but the toxic and side-effect of drug may be significant.