Phenotype of P63 knockdown zebrafish at 7dpf.

<p>(A) IHC for P63 protein on ESQE of zebrafish injected with control MOs. (B) IHC for P63 on ESQE of P63 knockdown zebrafish. (C) Normal zebrafish with pectoral fins (dorsal view). (D) Loss of pectoral fins in P63 knockdown zebrafish (dorsal view). (E) Normal ESQE. (F) Defective ESQE in a P63 knockdown zebrafish. (G) RT-PCR for P63 shows the un-spliced P63 transcript in MO injected zebrafish at 3, 5 and 7dpf. (H) Quantification shows a significant decrease of the number of ESQE cells in P63 knockdown zebrafish as compared with controls. All the pictures are dorsal side up. Base membrane of the squamous epithelium is marked with red dotted line and esophageal lumen is lined with black dotted line.</p

Identification of a non-keratinized stratified squamous epithelium in zebrafish upper digestive tract.

<p>(A-L) H&E staining of paraffin sections of zebrafish at 5, 6, 7 and 90 dpf shows the histogenesis of the squamous epithelium. A, D, G and J are the sagittal sections of the whole fish. B, E, H and K are magnifications of the areas in the yellow rectangles in A, D, G and J. Transverse sections show the histology of the squamous epithelium at 5, 6, 7 and 90 dpf (C, F, I, L). ISH for Krt5 (M, Q), IHC for P63 (N), IHC for Sox2 (O) and ISH for Pax9 (P, R) on transverse sections at 7dpf show the expression of esophageal genes in developing and adult zebrafish. All the pictures are dorsal side up. Base membrane of the squamous epithelium is marked with red dotted line and esophageal lumen is lined with black dotted line.</p

Expression of Pax9 in zebrafish ESQE and phenotypes of Pax9 knockdown zebrafish at 7dpf.

<p>(A) Normal zebrafish (Lateral view). (B) Pax9 knockdown fish with malformation of the lower jaw (Lateral view). (C) Alcian blue staining shows the orofacial cartilage of normal zebrafish (lateral view). (D) Alcian blue staining shows the malformed cartilage of Pax9 knockdown zebrafish (lateral view). (E) Normal ESQE in zebrafish. (F) Disorganized ESQE in a Pax9 knockdown zebrafish. (G) Krt5 ISH on normal zebrafish ESQE. (H) Krt5 ISH on Pax9 knockdown zebrafish ESQE. (I) Krt4 ISH on normal zebrafish ESQE. (J) Krt4 ISH on Pax9 knockdown zebrafish ESQE. (K) RT-PCR for Pax9 shows the un-spliced Pax9 transcript in MO injected zebrafish at 3, 5 and 7dpf. (L) Quantification shows a significant decrease of the number of ESQE cells in Pax9 knockdown zebrafish as compared with controls. (M) A schematic cartoon of normal zebrafish ESQE at 7dpf. (N) A schematic cartoon of Pax9 knockdown zebrafish ESQE at 7dpf. ep, ethmoid plate; ch, ceratohyal; mk, Meckel’s cartilage; tr, trabeculae. All the pictures are dorsal side up. Base membrane of the squamous epithelium is marked with red dotted line and esophageal lumen is lined with black dotted line.</p

Data_Sheet_1_Potential bioactive compounds and mechanisms of Fibraurea recisa Pierre for the treatment of Alzheimer’s disease analyzed by network pharmacology and molecular docking prediction.PDF

IntroductionHeat-clearing and detoxifying Chinese medicines have been documented to have anti-Alzheimer’s disease (AD) activities according to the accumulated clinical experience and pharmacological research results in recent decades. In this study, Fibraurea recisa Pierre (FRP), the classic type of Heat-clearing and detoxifying Chinese medicine, was selected as the object of research.Methods12 components with anti-AD activities were identified in FRP by a variety of methods, including silica gel column chromatography, multiple databases, and literature searches. Then, network pharmacology and molecular docking were adopted to systematically study the potential anti-AD mechanism of these compounds. Consequently, it was found that these 12 compounds could act on 235 anti-AD targets, of which AKT and other targets were the core targets. Meanwhile, among these 235 targets, 71 targets were identified to be significantly correlated with the pathology of amyloid beta (Aβ) and Tau.Results and discussionIn view of the analysis results of the network of active ingredients and targets, it was observed that palmatine, berberine, and other alkaloids in FRP were the key active ingredients for the treatment of AD. Further, Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis revealed that the neuroactive ligand-receptor interaction pathway and PI3K-Akt signaling pathway were the most significant signaling pathways for FRP to play an anti-AD role. Findings in our study suggest that multiple primary active ingredients in FRP can play a multitarget anti-AD effect by regulating key physiological processes such as neurotransmitter transmission and anti-inflammation. Besides, key ingredients such as palmatine and berberine in FRP are expected to be excellent leading compounds of multitarget anti-AD drugs.</p

Table_1_Potential bioactive compounds and mechanisms of Fibraurea recisa Pierre for the treatment of Alzheimer’s disease analyzed by network pharmacology and molecular docking prediction.XLSX

IntroductionHeat-clearing and detoxifying Chinese medicines have been documented to have anti-Alzheimer’s disease (AD) activities according to the accumulated clinical experience and pharmacological research results in recent decades. In this study, Fibraurea recisa Pierre (FRP), the classic type of Heat-clearing and detoxifying Chinese medicine, was selected as the object of research.Methods12 components with anti-AD activities were identified in FRP by a variety of methods, including silica gel column chromatography, multiple databases, and literature searches. Then, network pharmacology and molecular docking were adopted to systematically study the potential anti-AD mechanism of these compounds. Consequently, it was found that these 12 compounds could act on 235 anti-AD targets, of which AKT and other targets were the core targets. Meanwhile, among these 235 targets, 71 targets were identified to be significantly correlated with the pathology of amyloid beta (Aβ) and Tau.Results and discussionIn view of the analysis results of the network of active ingredients and targets, it was observed that palmatine, berberine, and other alkaloids in FRP were the key active ingredients for the treatment of AD. Further, Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis revealed that the neuroactive ligand-receptor interaction pathway and PI3K-Akt signaling pathway were the most significant signaling pathways for FRP to play an anti-AD role. Findings in our study suggest that multiple primary active ingredients in FRP can play a multitarget anti-AD effect by regulating key physiological processes such as neurotransmitter transmission and anti-inflammation. Besides, key ingredients such as palmatine and berberine in FRP are expected to be excellent leading compounds of multitarget anti-AD drugs.</p

Novel Resveratrol-Based Aspirin Prodrugs: Synthesis, Metabolism, and Anticancer Activity

Regular aspirin use has been convincingly shown to reduce the risk of colorectal cancer. However, long-term use of aspirin leads to gastrotoxicity. Herein, we designed and synthesized a novel class of resveratrol-based aspirin prodrugs to simultaneously release aspirin and resveratrol to attenuate the side effects caused by aspirin. Prodrug RAH exerted enhanced anticancer activities which are better than a physical mixture of aspirin and resveratrol as well as each individually. Metabolism of RAH in mice showed that the majority of RAH is decomposed to release resveratrol and aspirin or salicylic acid either in the intestine or after absorption. Mechanistic studies demonstrate RAH inhibits cell cycle arrest through downregulation of cyclins and induces apoptosis by activation of caspase-3 in cancer cells. These findings highlighted the improved anticancer properties of resveratrol-based aspirin prodrugs. RAH may represent novel and safe alternatives of aspirin for the purpose of daily use in the future

Novel Resveratrol-Based Aspirin Prodrugs: Synthesis, Metabolism, and Anticancer Activity

Regular aspirin use has been convincingly shown to reduce the risk of colorectal cancer. However, long-term use of aspirin leads to gastrotoxicity. Herein, we designed and synthesized a novel class of resveratrol-based aspirin prodrugs to simultaneously release aspirin and resveratrol to attenuate the side effects caused by aspirin. Prodrug RAH exerted enhanced anticancer activities which are better than a physical mixture of aspirin and resveratrol as well as each individually. Metabolism of RAH in mice showed that the majority of RAH is decomposed to release resveratrol and aspirin or salicylic acid either in the intestine or after absorption. Mechanistic studies demonstrate RAH inhibits cell cycle arrest through downregulation of cyclins and induces apoptosis by activation of caspase-3 in cancer cells. These findings highlighted the improved anticancer properties of resveratrol-based aspirin prodrugs. RAH may represent novel and safe alternatives of aspirin for the purpose of daily use in the future

Involvement of the Wnt pathway in the development of mouse esophageal epithelium.

<p>(A) X-Gal staining of E11.5 and E13.5 esophagi of BAT-GAL mice; (B) X-Gal staining of E13.5 esophagi of TOP-GAL mice. Es, esophagus; Tr, trachea; Ep, epithelium; Me, mesenchyme; Lu, lung.</p

Pathway changes in the three phases of mouse esophageal development.

<p>Note: ↑ and ↑ indicate up- or down-regulation, respectively.</p>a<p>Reference <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036504#pone.0036504-Rodriguez1" target="_blank">[4]</a>.</p>b<p>Reference <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036504#pone.0036504-Ohashi1" target="_blank">[43]</a>.</p

Esophageal hyperkeratosis due to <i>Nrf2</i> superactivation in <i>Keap1^−/−</i> esophagus.

<p>P7 esophagi of a wild-type mouse (A), a <i>Nrf2</i>^−/− mouse (B), a <i>Keap1</i>^−/− mouse (C), and a <i>Nrf2</i>^−/−<i>Keap1</i>^−/− mouse (D), were stained for H&E. Expression of Nrf2 (E–G), Pparγ/δ (H–J) and pAkt (K–L) were shown in the esophagi of P7 wild-type mouse, P7 <i>Keap1</i>^−/− mouse and adult wild-type mouse. Size bar = 50 µm.</p