Effects of Anthraquinones on Immune Responses and Inflammatory Diseases

The anthraquinones (AQs) and derivatives are widely distributed in nature, including plants, fungi, and insects, with effects of anti-inflammation and anti-oxidation, antibacterial and antiviral, anti-osteoporosis, anti-tumor, etc. Inflammation, including acute and chronic, is a comprehensive response to foreign pathogens under a variety of physiological and pathological processes. AQs could attenuate symptoms and tissue damages through anti-inflammatory or immuno-modulatory effects. The review aims to provide a scientific summary of AQs on immune responses under different pathological conditions, such as digestive diseases, respiratory diseases, central nervous system diseases, etc. It is hoped that the present paper will provide ideas for future studies of the immuno-regulatory effect of AQs and the therapeutic potential for drug development and clinical use of AQs and derivatives

Heritable Genome Editing with CRISPR/Cas9 in the Silkworm, <i>Bombyx mori</i>

<div><p>We report the establishment of an efficient and heritable gene mutagenesis method in the silkworm <i>Bombyx mori</i> using modified type II clustered regularly interspaced short palindromic repeats (CRISPR) with an associated protein (Cas9) system. Using four loci <i>Bm-ok</i>, <i>BmKMO</i>, <i>BmTH</i>, and <i>Bmtan</i> as candidates, we proved that genome alterations at specific sites could be induced by direct microinjection of specific guide RNA and Cas9-mRNA into silkworm embryos. Mutation frequencies of 16.7–35.0% were observed in the injected generation, and DNA fragments deletions were also noted. <i>Bm-ok</i> mosaic mutants were used to test for mutant heritability due to the easily determined translucent epidermal phenotype of <i>Bm-ok-</i>disrupted cells. Two crossing strategies were used. In the first, injected <i>Bm-ok</i> moths were crossed with wild-type moths, and a 28.6% frequency of germline mutation transmission was observed. In the second strategy, two <i>Bm-ok</i> mosaic mutant moths were crossed with each other, and 93.6% of the offsprings appeared mutations in both alleles of <i>Bm-ok</i> gene (compound heterozygous). In summary, the CRISPR/Cas9 system can act as a highly specific and heritable gene-editing tool in <i>Bombyx mori</i>.</p></div

Cas9/sgRNA-induced mutations at the <i>Bm-ok</i> locus in G₀<i>Bombyx mori</i>.

<p>(A) Schematic representation of <i>Bm-ok</i>. exons are shown as boxes and arrows represent the primers used to amplify the target regions. Target site locations (<i>Bm-ok</i>-tar1 and <i>Bm-ok</i>-tar2) are underlined and PAM sequences are shown in red. (B) Representative chromatograms of PCR-product sequencing in G₀ silkworms in which indel mutations are present. (C) Sequences of indel mutations at the targeted <i>Bm-ok</i> locus in G₀ silkworms. The target sites are highlighted in green and PAM sequences are shown in red. Deletions are indicated by hyphens and insertions are shown in lowercase red letters. The indel mutation type is noted to the right (+, insertion; -, deletion). (D) Images indicating characteristic <i>Bm-ok</i> mosaic mutation phenotypes: (a) Fifth instar G₀ wild-type (P50) (left) and mosaic mutant (right) individuals; (b–d) Magnified images of wild-type and mutant individuals. The scale bars represent 1 mm.</p

Mutations induced by Cas9/sgRNA microinjection in G₀.

<p>Mutations induced by Cas9/sgRNA microinjection in G₀.</p

G₁ germline mutations from Crossing Strategy 2.

<p>G₁ germline mutations from Crossing Strategy 2.</p