ABSTRACT
Objective: To express DNA-binding protein (DBP) of human adenovirus (HAdV) type 7 using the prokaryotic expression system, and product anti-HAdV-7 DBP rabbit polyclonal antibody. Methods: The HAdV-7 DBP gene was synthesized and cloned into prokaryotic expressing vector pET30a, and the recombinant plasmid was transformed into E. coli BL21 (DE3) competent cell. The recombinant protein DBP was expressed by induced Isopropyl-beta-D-thiogalactopyranoside (IPTG) and purified with Ni-NTA affinity column. The titer of anti-DBP polyclonal antibody produced in immunized rabbit was measured by indirect ELISA, and the specificity of the antibody was identified by Western blotting and indirect immunofluorescence assay (IFA). In addition, purified rDBP was used as coating antigen for indirect ELISA assay to detect specific IgM and IgG antibodies against DBP in the serum of children infected with HAdV. Results: The HAdV-7 DBP plasmid was constructed successfully. The purified recombinant DBP was more than 95% after purification. The titer of polyclonal antibody was 1∶1 024 000. The polyclonal antibody showed high specificity in vitro using Western blotting and IFA. The positive rate of specific anti-DBP IgM and IgG antibody in acute-phase serum samples collected from children infected with HAdV were 50.0% (19/38) and 63.2% (24/38), respectively, using indirect ELISA. Conclusion: In summary, the HAdV-7 rDBP is expressed using prokaryotic expression system, and the recombinant HAdV-7 DBP protein and the anti-DBP rabbit polyclonal antibody with high titer are prepared.
Subject(s)
Animals , Rabbits , Adenoviruses, Human/genetics , Antibody Specificity , Blotting, Western , DNA-Binding Proteins/metabolism , Enzyme-Linked Immunosorbent Assay , Escherichia coli/genetics , Immunoglobulin GABSTRACT
Objective To explore the effect of medical assessment on the rehabilitation training of children with autism spectrum disorders (ASD). Methods There were 67 children with ASD selected from special education schools .Their developmental level , ability of social life and social communication were evaluated by child health care physicians .There were 48 children classified as intervention group:individualized rehabilitation training targets were designed based on the assessment results and individualized rehabilitation training program was carried out .The other 19 children were served as controls and routine rehabilitation training was carried out .All children were reevaluated by physicians with the same methods after training for one year . Results The adaptability , language level and communication ability of children in the intervention group improved better than the control group , with statistically significant difference. Conclusion It is beneficial to develop training programs for children with ASD on the basis of medical assessment which can promote the development level of ASD children , social life ability and communication ability .
ABSTRACT
[Objective] To explore the relationship between children ’ s adaptation to kindergarten and their temperament type . [ Methods ] Initial survey: A self-made questionnaire was used to collect the relevant information.After that,a behavior screening questionnaire (BSQ) for 3 to 7 year old children was applied to assess the temperament type of those who were new to the kindergarten .Longitudinal study:Changes in children ’ s behavior in the kindergarten and at home were recorded by using a self-made obser-vation sheet in order to know their adaption situation .All participants were followed up 5 weeks. [ Results] Inadaptation to kindergarten was found to be obvious among children who were new to the kin-dergarten and the time needed for their adaption was one month .Adaption to kindergarten in children with positive temperament was much better than those with negative temperament (P<0.05). [Conclusion] The temperament type of children is correlated to their kindergarten adaption .