Establish and optimization of inter-simple sequence repeat PCR reaction system of Gnaphalium affine / 药学实践杂志

Objective To provide the experimental basis for the subsequent genetic diversity research through establishing and optimizing the inter-simple sequence repeat PCR (ISSR-PCR) reaction system of Gnaphalium affine. Methods The single-factor experimental method and full experimental method were used to optimize the ISSR-PCR reaction system of Gnaphalium affine. Under the optimal system, after screening primers and corresponding annealing temperatures, the systematic feasibility was verified. Results The optimal ISSR-PCR reaction system was consisted of 10 μl Premix Taq DNA polymerase, 0.3 μmol/L primer, 10 ng DNA template, and sterilized water added to 20 μl. Finally, 10 primers were screened from 100 universal primers, and verification results indicated the system had high stability, good reproducibility, and the selected primers had good polymorphism. Conclusion The ISSR-PCR amplification system of Gnaphalium affine was established for the first time and the primers with appropriate annealing temperatures were filtered out, which provided a reference for the subsequent genetic diversity research of Gnaphalium affine.

Study on synthesis and cyto-toxicity of polyethylene glycol-b-polylysine as poten-cial miRNA carrier / 药学实践杂志

Objective To synthesize miRNA carrier PEG-b-PLL and to testify the stability , encapsulation efficiency and cyto-toxicity of its complexes .Methods H1 NMR was used to determine the degree of polymerization of PLL , 4%agarose gel electrophore-sis was used to determine entrapment of the polyer to the miRNA;then dynamic light scattering ( DLS) was used to measure the hydro-dynamic parameter such as size , polydispersity index ( PDI) and zeta potential of the polyplexes .The entrapment efficiency was deter-mined by ultraviolet-visible spectrophotometer , and finally , the cyto-toxicity of PEG-b-PLL was evaluated by CKK-8 kit with K562 cell lines.Results The characteristics indicated polyplexes prepared by PEG-b-PLL and miRNA fulfill the demand of being the gene carri-er of miRNA because of low cyto-toxicity , high encapsulation efficiency and stability .Conclusion The miRNA carrier PEG-b-PLL had good character and low cyto-toxicity.It showed considerable potential as an efficient miRNA carrier .