Objective To produce anti-thrombomodulin antibodies.Methods Using genetic immunization: Eukaryotic expression plasmid pcDNA3.1/TM(LEO),encoding all the extracellular domain of human thrombomodulin and signal peptides but lacking the transmembrance and cytoplasmic domains was constructed, which recombinant thrombomodulin was secreted soluble product. The plasmid was isolated from large-scale bacterial cultures by treatment with alkali and SDS, purified by precipitation with polyethylene Glycol (PEG). Recombinant plasmid was injected into tibial muscle of BALB/c mice. The productions of TM and anti-TM have been detected. Results The positive of RT-PCR and expressed TM identified the function of the recombinant plasmid. The pcDNA3.1/TM(LEO) induced higher titer of anti-TM. The antibody titer peaked between the 5th and 7th injection with a titer of 1∶8 000 detected by cell-ELISA coated with EVC-304. Specificity has been identified by western blot and immunohistochemistry.Conclusion The production of antibody through genetic immunization was a feasible method due to the difficulties in obtaining and purification of natural thrombomodulin.

AIM and METHODS: The purpose of the study was to characterize the time-related effect of Danazol therapy on endometriosis explant using the rat model. Endometriosis was induced in mature female rats. One group of treated animals as well as controls were sacrificed at 2,4,6 and 8 weeks after treatment at which time the explant was evaluated. RESULTS: Explant volume was significantly reduced in all treatment groups, and the effect was more significant in animals treated for 4 weeks than those treated for only 2 weeks. CONCLUSION: Danazol treatment can cause gradual regression of endometrial explant in a time-related manner.