ABSTRACT
Objective:To demonstrate the polymorphism of α chain of bovine major histocompatibility complex(BoLA)classⅠmolecule and domain binding constant chain(Ii).Methods:Total 75 BoLA Iα genes were obtained from three Huaibei cattle and analyzed by molecular biology software;the genes of typical BoLA Iα domains and Ii were cloned,and then inserted into prokaryotic expression plasmid.After induced protein expression;the domains of BoLA Ⅰα chain binding to Ii were detected by pull-down meth-od and Western blot.The recombinant eukaryotic expression plasmids were constructed and the co-localization of BoLA Iα segments with Ii was observed by laser confocal microscopy.Results:Firstly,it was found that there were at least 5 kinds of BoLA Iα in the cloned gene sequence,which were highly polymorphic and they were mainly distributed in the antigen peptide binding region(PBR)of BoLA Ⅰ(α1α2)and cytoplasmic region.Secondly,the prokaryotic recombinant plasmids containing BoLA Ⅰα1α2α3,BoLA Ⅰα1α2 or BoLA Ⅰα 3 were constructed,then they were respectively induced to express and purified,in which,the BoLA Ⅰα1α2α3 and BoLA Ⅰα1α2 had the activity of binding to Ii.Finally,in 293T cells BoLA Ⅰα1α 2α3 or BoLA Ⅰα1α2 was found that could co-localize with Ii,while a single BoLA Ⅰα3 could not.Conclusion:BoLA Ⅰα gene is highly polymorphic.BoLA Ⅰα1α2 is a func-tional fragment that binds to Ii and co-locates intracellular.
ABSTRACT
Objective:To determine the content of self-manufactured and imported lurasidone hydrochloride tablets in order to e-valuate their internal qualities. Methods:The determination of lurasidone hydrochloride tablets was performed by HPLC. The HPLC system consisted of a Waters C8 column (250 mm × 4. 6 mm, 3. 5 μm) and the mobile phase of 0. 05 mol·L-1 phosphate buffer solu-tion (pH 3. 0)-acetonitrile(60∶40), the detection wavelength was 230 nm, the flow rate was 1. 2 ml·min-1 and the column tempera-ture was 40℃, and the injection volume was 20μl. Results:The linear range of lurasidone hydrochloride was 0. 100 8-0. 806 4 mg· ml-1(r=0. 999 5). The average recovery was 99. 95% with RSD of 0. 31%(n=9). Conclusion:The method is simple, rapid, ac-curate, and reliable. The method can determine lurasidone hydrochloride tablets satisfactorily. According to the results, there are few differences among the self-manufactured and imported lurasidone hydrochloride tablets.