Objective To isolate specific humanized anti-D-dimer scFv(single chain Fv) antibody from scFv phage libraries. Methods Isolate anti-D-dimer positive clones from Tomlinson I + J phage libraries by three rounds of panuing, then sequence monoclonal genes by bideoxy-mediated chain termination and express soluble scFv antibody; Pick out anti-D-dimer antibodies with high specificity and affinity by ELISA.Results After three rounds of selection from human scFv phage libraries Tomlinson I and J, 38 monclonal specific anti-D-dimer scFv fragments were selected. By polyclonal and monoclonal phage ELISA and gene sequencing, 20 different full-length monoclonal scFv phages were identified, the result of soluble scFv ELISA showed that 20 full-length monoclonal scFv were expressed smoothly. According to the result of soluble scFv ELISA, in 5 scFv antibodies with high value of A450 selected, 3 scFv antibody fragments showed high specific and affinity. Conclusion Antibody phage display was an effective, rapid method to isolate anti-D-dimer antibodies with high specificity and affinity.

Objective To evaluate the effect of co -administration of the loop diuretic furosemide (Fur) and kanamycin (KM ) in the rats in order to establish a reliable animal model of sensorineural hearing loss .Methods Thirty -two Sprague-dawley (SD) rats were randomly divided into 4 groups with 8 rats in each group .Rats in group A ,B and C received a single intraperitoneal injection of kanamycin sulfate (KM ,500 mg/kg) ,followed by an-other intraperitoneal injection of furosemide (Fur ,0 .2 ml/kg ) 30 minutes later .While ,rats in control group D re-ceived same doses of normal saline .Auditory brain responses (ABRs) were recorded at 1 ,7 and 14 day after the in-jections ,which were group A ,group B and group C ,respectively .Hair cell loss ,the spiral ganglions and microstruc-ture were observed by immunofluorescence and scanning electron microscopy .Results The ABR thresholds in group A ,B and C were significantly higher than that of in control group D (P<0 .05) .There were no differences of ABR thresholds among group A ,B and C(P>0 .05) .Immunofluorescence and scanning electron microscopy showed obvi-ous hair cell loss in the basal turn in each kanamycin groups ,with cilia disarrangement and fusion ,compared to the same areas in animals from the control group .The expression of cleaved caspase -3 significantly increased in each experimental group than that of in the control group(P<0 .05) .Conclusion Co -administration of furosemide and kanamycin intraperitoneally induces profound hearing loss in rats and is an effective method of establishing acute hearing loss model in animal experiments .