Objective: To express recombinant human angiostatin for further application in clinic. Methods: The complete encoding eDNA of human angiostatin was isolated from human embryo liver with RT-PCR and expressed in secretory Pichia expression system. Recombinant human angiostatin was purified with heparin sepharose chromatography and its activity was determined in chick embryo chorioallantoic membrane (CAM) and wound healing assays. Results: Expressed in large quantity (yield=5 mg/L) and purified with heparin sepharose, recombinant angiostatin was showed to have a molecular weight of 43 kD in SDS-PAGE and potently inhibit angiogenesis and wound healing. Conclusion: Recombinant human angiostatin was expressed efficiently in a biologically active form.

Objective: To express recombinant human soluble fibroblast growth factor receptor 1 ( sFGFR1) and study its antagonistic activity on FGF. Methods: Human sFGFR1 cDNA, isolated from human lung fibroblast cells with RT-PCR was confirmed by DNA sequencing and cloned into pYEX4T-1 yeast expression vector. The recombinant sFGFR1 was expressed in DY150 yeast cells and the product was identified by SDS-PAGE and Western blot. The activity of recombinant sFGFR1 was detected in N1H3T3 proliferation inhibition assay. Results: GSF-sFGFR1 fusion protein was expressed in yeast cells and was observed as a band of 60 Id) on a SDS- PAGE gel and by Western blot. The recombinant fusion protein was also found to be able to suppress FGF-induced proliferation of NIH3Th cells. Conclusion: Recombinant human GST-sFGFR1 fusion protein was expressed in yeast efficiently and showed natural biological activities.

Objective:To study the clinical application potential of aFGF and try to produce aFGF as gene engineering medicine.Methods:The complete encoding cDNA of human aFGF was isolated from human lung fibroblast with RT PCR.Recombinant human aFGF was expressed in secretory pichia expression system and purified with heparin sepharose chromatography.The activity of aFGF was detected in NIH3T3 proliferation assay,chick embryo chorioallantoic membrane assay(CAM) and wound healing assay.Results:The recombinant aFGF was expressed in large quantity(yield=12 mg/L) and was capable to promote proliferation of NIH3T3,angiogenesis and wound healing.Furthermore,those activities of aFGF could be agonized by recombinant FGFR extracellular domain.Conclusion:Recombinant human aFGF was expressed efficiently and possessed natural biological activities.

Objective:To study the differences of bleomycin-induced pulmonary fibrosis in mice induced by intraperitoneal injection and intratracheal instillation of bleomycin.Methods:ICR mice (male,8 w of age,18 to 22 g bodyweight) were used.①ICR mice were randomly divided into three groups.In the group P,bleomycin was injected intraperitoneal five times in a dose of 40 mg/kg,and in the group I,bleomycin was instilled intratracheally in a dose of 5 mg/kg.The mice were killed at 14,28 or 40 day.②ICR mice were randomly divided into four groups : bleomycin was injected intraperitoneal three,four or five times in a dose of 40 mg/kg,and in the group control ,bleomycin was injected intraperitoneal in a dose of 200 μl for five times.The mice was killed at 28 or 40 day.The pathological changes and symptoms were observed.Results:① The weight of the mice given blemycine were lost after injection.Symptoms were more serious in group P than those of group I.The pulmonary coefficient of group P was more than that of group I.At 28 day after injection,fibrosis was widely and stably formed mainly in around the bronchia and bronchioles,especially,near the pulmonary hilar area,in the group I mice,however,the same changes were mainly seen under the pleura or perivascular pulmonary tissues in the mice of group P.The pathological score of pulmonary fibrosis was different between those two groups.②Different dose of bleomycine induced different change of the mouse's weight.The most of all of the three group were five time injection; The lung index of five time injection of bleomycine was the most.The pulmonary fibrosis mouse model was made successfully.After comparising all of the data,we found intraperitoneal of bleomycine five times was the more convenient method.Conclusion:The sites of pulmonary fibrosis in the mice are different between the mice induced by intraperitoneal injection or intratracheal instillation of bleomycin.Intraperitoneal injection five times is a more convenient method to make pulmonary fibrosis.