Outbred genome sequencing and CRISPR/Cas9 gene editing in butterflies.

Butterflies are exceptionally diverse but their potential as an experimental system has been limited by the difficulty of deciphering heterozygous genomes and a lack of genetic manipulation technology. Here we use a hybrid assembly approach to construct high-quality reference genomes for Papilio xuthus (contig and scaffold N50: 492 kb, 3.4 Mb) and Papilio machaon (contig and scaffold N50: 81 kb, 1.15 Mb), highly heterozygous species that differ in host plant affiliations, and adult and larval colour patterns. Integrating comparative genomics and analyses of gene expression yields multiple insights into butterfly evolution, including potential roles of specific genes in recent diversification. To functionally test gene function, we develop an efficient (up to 92.5%) CRISPR/Cas9 gene editing method that yields obvious phenotypes with three genes, Abdominal-B, ebony and frizzled. Our results provide valuable genomic and technological resources for butterflies and unlock their potential as a genetic model system.

Aqueous extract of Huang-lian induces apoptosis in lung cancer cells via P53- mediated mitochondrial apoptosis

The current study was designed to evaluate the activity of the aqueous extract of Huang-lian, and the main apoptosis pathway induced by the extracts of Huang-lian was detected on lung cancer. Antiproliferactive activity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and TUNEL methods against human lung cancer cells (A549). Huang-lian regulated the Bcl-2 family protein-mediated mitochondrial pathway via p53 and was detected by western blot. It increased the activation of caspase-3 and caspase-3 cleavage increased as the time increased. These results suggested that Huang-lian regulated the Bcl-2 family protein-mediated mitochondrial via p53 pathway, suggesting that Huang-lian should be further investigation as a natural agent for treating and preventing cancer.