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Objective To construct high-capacity ribosome display single-chain Fv library for selection of high affinity ScFv antibody.Methods We isolate human lymphocyte from peripheralblood(2 normal,3 gastric cancer,3 colonic cancer,1 pancreatic cancer,each 5 mL and 2 newborn,each 2 mL)and extract RNA for cloning whole human heavy chain and light chain gene by RT-PCR.VH and VL were rearranged randomly by SOEing(splicing by overlap extension,SOEing).Finally,the elements for in vitro screening such as T7 promoter and ribosome binding site were introduced while the SOEing products were amplified.Moreover,ribosome display template were verified by blue/white screening and further sequencing.Results We successfully constructed ribosome display ScFv library with a volume of 1.1?1013.Conclusion The construction of high-capacity ScFv library shed light on multiple therapeutic ScFv screening.
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AIM: To investigate the relationship between expression of Bmi-1(B cell-specific MLV integration site-1) in gastric cancer and its clinicopathologic significance.METHODS: 146 surgical patients with gastric carcinoma were followed up at least 2 years.Expression of Bmi-1 protein was examined by immunohistochemistry in their archival paraffin embedded tissue specimens.RESULTS: The intensive positive rate of Bmi-1 expression in gastric cancer was 67.8%(99/146).Expression of Bmi-1 was highly correlated with tumor size,clinical stage,lymph node metastasis and T classification(P0.05).The survival rate in the patients with Bmi-1 expression was much lower than that in those patients without Bmi-1 expression(P
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The Leishmania Donovani, in bone morrow of 1 case Kala-Azar was observed with electron microscope. The results showed that the ultrastructure of the Leishmania Donovani was similar to that of other Leishmanial species, however, there were some differences in the size of parasites, microtubule intervals and numbers.