ABSTRACT
Objective: To construct the cell models of TNF-α rs1800620 (SNP1) and TNF-α rs4645843 (SNP2) stable transgen-ic lines and study the effects of SNPs on the content of TNF-α in LX-2 cell line. Methods: TNF-α SNP1/TNF-α SNP2 mutant plasmid was synthesized by wild-type TNF-α targeting and then transfected into LX-2 cell line after lentivirus packaging. RT-qPCR and Western blot were used to detect the mRNA and protein levels of TNF-α in each recombinant cell. Results: The gene sequencing showed that plasmids were successfully constructed. Photographs from an inverted microscope showed that the recombinant cells had stable expres-sion of GFP. The results of RT-qPCR and Western blot showed that compared with the wild group, SNP1 and SNP2 could significantly reduce the mRNA (P<0. 05) and protein (P<0. 05) levels of TNF-α. Conclusion: The recombinant cells stably expressing TNF-α/TNF-α SNP1/TNF-α SNP2 LX-2 are constructed. SNP1 and SNP2 can significantly reduce the mRNA and protein levels of TNF-α in LX-2 cell line.
ABSTRACT
Objective:To prepare loratadine nanosuspension lyophilized powder (LTD-NLP) and investigate the dissolution and stability. Methods:LTD-NLP was prepared by an anti-solvent precipitation method; the particle size and morphology were characterized by a laser nanometer particle sizer and a transmission electron microscope (TEM). The optimal lyophilized protective agent was screened out; the dissolution and stability of LTD-NLP were determined by HPLC. Results:The average particle size of LTD-NLP was 175.6 nm with PDI of 0.200, and the zeta potential was -56.3 mV. The shape of LTD-NLP was spherical. The nanosuspension lyophilized with 10% sucrose presented optimal properties. The dissolution of LTD-NLP was greater than raw material, and LTD-NLP had good stability at 4℃. Conclusion:The anti-solvent precipitation method for LTD-NLP is simple,and the product is expected to become a new preparation of loratadine.