Huang W.-Y.,Changchun Institute Of Biological Products Coltd |
Long F.-Y.,Changchun Institute Of Biological Products Coltd |
Yang C.-C.,Changchun Institute Of Biological Products Coltd |
Zhu Y.-R.,Changchun Institute Of Biological Products Coltd |
And 2 more authors.
Chinese Journal of Biologicals | Year: 2016
Objective To construct the shRNA lentivirai vector for filamin A anil determine its knockout efficiency in HepG2 and Huh7 cells. Methods The knockout gene fragment was synthesized and inserted into the shRNA lentivirai vector GV298 with a cherry protein reporter. The constructed recombinant plasniids shRNA-Filaminl and shRNA-Filamin2 were transfected into HepG2 and Huh7 cells respectively. The expression of cherry protein was observed liy fluorescent microscopy, and the knockout efficiency of filamin A was determined by Western blot. Results Sequencing results proved that the FLNa shRNA lentivirai vectors for filamin A were constructed correctly. Strong cherry fluorescence was observed in HepG2 and Huh7 cells 24 h after transfectioti. However, the knockout efficiencies of shRNA-Filaminl and shRNA- Filarnin2 were 45. 4% and 63. 77c in HepG2 cells, and 76. 5% and 78. 3% in Huli7 cells, 48 h after transfection, respectively. Conclusion Tlie FLNa shRNA lentivirai vectors for filamin A were successfully constructed, of which the knockout efficiencies were high in IIepG2 and Huh7 cells. Filamin A was able to interact with the receptor proteins of various viruses, thus participate in the replication cycle of the virus. It provided a new approach for study on the virus replication and the pathogenic mechanism.
Miao L.,Changchun Institute Of Biological Products Coltd |
Ding L.-L.,Changchun Institute Of Biological Products Coltd |
Li C.-Y.,Changchun Institute Of Biological Products Coltd |
Zhao B.,Changchun Institute Of Biological Products Coltd |
And 4 more authors.
Chinese Journal of Biologicals | Year: 2014
Objective To evaluate the long-term toxicity of freeze-dried rabies vaccine (Vero cells) for human use in machins after repeat intramuscular injection. Methods Twenty-four machins were randomly divided into negative control, auxiliary material control, as well as low (1 dose / time ) and high (5 doses / time) dose groups, six for each, with equal genders. The low and high doses were 1 and 5 times of those for human use in clinic respectively. The machins in each group were injected i.m. on days 0, 3, 7, 14, 28 and 42 respectively, and observed for 4 weeks after the last injection for general clinical indexes, bodyweight, body temperature, electrocardiogram (ECG), ophthalmology, hematology, blood biochemistry, electrolyte index, urine index, immune index, bone marrow smear as well as pathological changes in organs and tissues. Results The general state of animals in various groups were normal, while no visual adverse reactions were observed in injection site. No regular changes of toxicological significance were observed in bodyweight, body temperature, ECG, blood cell count, coagulation function, blood biochemistry, ophthalmology, routine urine examination, peripheral blood T lymphocyte subset distribution, serum IL-2 and IFNΥ levels, bone marrow or organ coefficient. The serum specific antibody level against rabies virus increased significantly in low and high dose groups, while protective (more than 0. 5 IU / ml) neutralizing antibodies were induced. Mild irritant changes in injection site were observed in auxiliary material control as well as low and high dose groups, which resolved after a convalescent period of 4 weeks. Conclusion The non-toxic reaction dosage of freeze-dried rabies vaccine for human use was less than 5 doses / time, while local irritant reaction in injection site should be noted in clinic.