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Studying On Treatment Hemangioma With Calreticulin-like Protein

Posted on:2012-08-11Degree:MasterType:Thesis
Country:ChinaCandidate:Z H ZhangFull Text:PDF
GTID:2254330425455831Subject:Biochemistry and Molecular Biology
Abstract/Summary:PDF Full Text Request
A hemangioma is an abnormal hyperplasia of blood capillary of children, a common and frequently-occurring disease, whose effective treatment has also not been processed presently. However, the angiogenesis inhibitor is kind of protein could specifically inhibit the growth of endothelial cells. And the treatment trial for hemangioma using Careticulin-like protein (CLP) were carried out in this study.The CLP gene has been amplified by SOE PCR. And then the expression vector including CLP gene, pPIC9K-careticulin-like, were established, which were transfered into the Pichia pastoris genome by electroporation. Subsequently high-copy recombinants were screened by G418resistance and the engineering strain GS115was selected. The GS115recombinant protein has been abundantly expressed through ferment and purified using ion exchange method. Succedent test showed the purified target protein, CLP, was active to inhibit angiogenesis.The CLP were able to induce the necrosis of hemangioma, mainly manifested in cell shrinkage.Cock of the wattle is a natural blood vessel tumor model. In this study,60-day-old rooster’s wattle vascular tumor model, the use Careticulin-like protein in hemangioma test. In the experimental group on the right wattle daily injections Careticulin-like protein20μ g, the control group injected with a daily volume of the left wattle of PBS.14days after the drug treatment group, the average length of wattle1.90cm, wattle length of the control group average of2.52cm, showed a significant difference (P <0.001); medication side wattle test group the average number of blood vessels as a/100field, the control group for a number of blood vessels/100vision, number of blood vessels between the two groups was significant difference (P<0.001); medication tissues of the control group, normal cells, and vascular supply of nutrients due to lack of experimental group, the significant local tissue necrosis medication. The average weight of the experimental group increased chicken275g/only in the control group gained an average growth of chicken325g/only, changes in body weight between the two groups no significant difference (0.1<P<0.25).These forementioned results suggested the CLP could effectively inhibit the proliferation of vascular tumor cells, new-growth of vessel in hemangioma model and expansion of hemangioma.With the development of drugs for the treatment of hemangioma prospects.
Keywords/Search Tags:Careticulin-like protein, hemangioma, Pichia pastoris, highdensity fermentation
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