Image1_Dihydromyricetin-Encapsulated Liposomes Inhibit Exhaustive Exercise-Induced Liver Inflammation by Orchestrating M1/M2 Macrophage Polarization.TIF

Exhaustive exercise (EE) induced hepatic inflammatory injury has been well reported. Dihydromyricetin (DHM) has shown anti-inflammatory bioactivity and hepatoprotective effects but is limited by poor bioavailability. Here, high-bioavailability DHM-encapsulated liposomes were synthesized and explored for their therapeutic potential and regulatory mechanisms in a hepatic inflammatory injury model. The animal model was established by swimming-to-exhaustive exercise in C57BL/6 mice, and the anti-inflammatory effects were detected after administration of DHM or DHM liposome. NIR fluorescence imaging was used to assess the potential of liver targeting. The DHM liposome-induced macrophage polarization was measured by flow cytometry ex vivo. The anti-inflammatory mechanism of DHM was studied in cell line RAW264.7 in vitro. Liposome encapsulation enhanced DHM bioavailability, and DHM liposome could alleviate liver inflammation more effectively. Moreover, DHM liposome targeted hepatic macrophages and polarized macrophages into an anti-inflammatory phenotype. The SIRT3/HIF-1α signaling pathway could be the major mechanism of DHM motivated macrophage polarization. Our study indicates that DHM liposomes can alleviate liver inflammation induced by EE through sustained releasing and hepatic targeting. It is a promising option to achieve the high bioavailability of DHM. Also, this study provides new insights into the regional immune effect of DHM against inflammation.</p

Interactions between Zfpm2 variants and Gata4 assessed by GST pull-down assays.

<p>Coomassie blue staining indicated the amounts of GST fusions pulled down in the assays. The relatively band intensities compared with the wild type are shown in the bottom. Molecular mass standards (kDa) are indicated at the left. Lane 1, GST; Lane 2, GST-Zfpm2 wild type; Lane 3, GST-Zfpm2 p.M703L; Lane 4, GST-Zfpm2 T843M; Lane 5, GST-Zfpm2 E30G; Lane 6, GST-Zfpm2 I227V; Lane 7, GST-Zfpm2 K737E; Lane 8, GST-Zfpm2 S402R; Lane 9, GST-Zfpm2 M544I.</p

Inhibition of autophagy or over-expression of β-catenin attenuates the effects of resveratrol on BCSCs.

<p>(A) Western blot analysis of β-catenin and cyclin D1 in BCSCs treated with different dose of resveratrol for 24 h. (B) Immunohistochemistry detection of β-catenin and cyclin D1 in xenografted breast tumors. (C) The effects of resveratrol treatment for 24 h on ALDH-positive populations in SUM159 cells and SUM159 cells co-treated with 10 µM CQ (CQ-SUM). (D) The effects of resveratrol treatment for 24 h on autophagy in BCSCs, BCSCs co-treated with 10 µM CQ (CQ-CSCs) and BCSCs transfected with pcDNA3-S33Y β-catenin (TC_β-catenin). (E) The effects of resveratrol treatment for 24 h on cells viability in BCSCs, CQ-CSCs and TC_β-catenin cells. (F) Western blotting analysis of β-catenin and cyclin D1 in TC_β-catenin cells treated with different dose of resveratrol for 24 h. Data are presented as mean ± SD (<i>n</i> = 3). <b>^*</b><i>P</i><0.05, <b>^**</b><i>P</i><0.01 compared with the control (0 µM); <b>^#</b><i>P</i><0.05, <b>^##</b><i>P</i><0.01 compared with corresponding group in CSCs.</p

Novel Missense Variants of ZFPM2/FOG2 Identified in Conotruncal Heart Defect Patients Do Not Impair Interaction with GATA4

<div><p>Conotruncal heart defect (CTD) is a complex form of congenital heart disease and usually has a poor prognosis. <i>ZFPM2/FOG2</i> encodes a transcription cofactor that interacts with GATA4 to regulate cardiac development. This regulation has been established in knockout mouse models that display a range of heart malformations, especially CTD. Although previous studies have identified several missense variants in <i>ZFPM2/FOG2</i> that may cause CTD, it remains unclear whether they are involved in CTD pathogenesis because the study populations were limited and the functional status was unknown. In this report, we screened a larger CTD population, which comprised 145 tetralogy of Fallot (TOF), 37 double-outlet ventricle outflow (DORV), and 18 transposition of the great artery (TGA), to investigate exon mutations as well as copy number variations in <i>ZFPM2/FOG2</i>. Four variants (p.V339I in one DORV, p.A426T in one DORV, p.M703L in three TOF, p.T843M in one TOF) were found in six patients, of which two are reported here for the first time. No copy number variations of the gene were detected. GST pull-down assays demonstrated that all potentially deleterious variants, including those previously reported, did not impair the interaction with GATA4, except for variant p.M544I and p.K737E, which subtly impaired the binding. Thus, these missense variants may be involved in other mechanisms underlying CTD or may be unrelated to CTD occurrence.</p></div

Resveratrol induces autophagy in BCSCs.

<p>(A) Resveratrol treatment (40 µM) for 24 h increased the number of autophagosomes in BCSCs from SUM159 and MCF-7, determined by TEM analysis. (B) Western blotting detection of LC3-II, Beclin1 and Atg 7 in BCSCs treated with different dose of resveratrol for 24 h. (C) GFP-LC3-II puncta formation assay in BCSCs treated with resveratrol for 24 h. Data are presented as mean ± SD (<i>n</i> = 3). <b>^*</b><i>P</i><0.05, <b>^**</b><i>P</i><0.01 compared with the control (0 µM).</p

Resveratrol inhibits BCSCs <i>in vivo</i>.

<p>(A) Resveratrol injection (100 mg/kg/d, i.v.) effectively inhibits the growth of xenografted tumors of SUM159 cells in NOD/SCID mice. (B) Resveratrol administration significantly reduced the ALDH-positive populations in tumor cells dissociated from xenograft in NOD/SCID mice. Data are presented as mean ± SD (<i>n</i> = 6). <b>^*</b><i>P</i><0.05, <b>^**</b><i>P</i><0.01 compared with the control.</p

Locations of mutations identified and sequence alignments in ZFPM2/FOG2.

<p>Top, the N-terminal transcriptional repression domain is indicated in red; the eight zinc-finger motifs are represented by blue; the nuclear localization signal is indicated in yellow; the putative CtBP-binding site is represented by green. Variants identified in this study are shown underneath the protein structure; variants reported in previous studies are shown above. Bottom, multiple alignment of partial amino acid sequences of human ZFPM2/FOG2 and its homologs from other species. Variant residues are boxed. Accession numbers of the sequences used are as follows: Homo, NP_036214.2; Pan, XP_001158075.1; Canis, XP_539118.2; Mus, NP_035896.1; Rattus, XP_235253.4; Gallus, XP_418380.2; Danio, NP_001034724.1.</p

Sequencing chromatograms of four heterozygous missense variants.

<p>(A) Each arrow indicates the position of the nucleotide variant that results in the substitution of wild-type Val with Ile at codon 339. (B) Substitution of wild-type Ala with Thr at codon 426. (C) Substitution of wild-type Met with Leu at codon 703. (D) Substitution of wild-type Thr with Met at codon 843.</p

Data_Sheet_2_Long-term high loading intensity of aerobic exercise improves skeletal muscle performance via the gut microbiota-testosterone axis.docx

Exercise is reported to play a crucial role in skeletal muscle performance. However, the underlying mechanism is still unknown. Thus, we investigated the effect of high-intensity aerobic exercise on skeletal muscle performance. In this study, the male C57BL/6J mice were accepted by high-intensity aerobic exercise for 8 weeks to establish an exercise model. It was observed that high-intensity aerobic exercise markedly affected the expression of genes in skeletal muscle. Moreover, high-intensity aerobic exercise significantly improved skeletal muscle grip strength and serum testosterone levels. HE staining showed that the cross-sectional area (CSA) of the skeletal muscle was successfully increased after 8 weeks of high-intensity aerobic exercise. Additionally, we found that high-intensity aerobic exercise changed gut microbiota structure by altering the abundance of Akkermansia, Allobaculum, and Lactobacillus, which might be related to testosterone production. However, the beneficial effects disappeared after the elimination of the gut microbiota and recovered after fecal microbiota transplantation (FMT) experiments for 1 week. These results indicated that the beneficial effects of high-intensity aerobic exercise on skeletal muscle were partly dependent on the gut microbiota. Our results suggested that long-term high loading intensity of aerobic exercise could improve skeletal muscle performance, which was probably due to the gut microbiota-testosterone axis.</p

Table_1_Interaction between the flagellum of Candidatus Liberibacter asiaticus and the vitellogenin-like protein of Diaphorina citri significantly influences CLas titer.DOCX

Huanglongbing (HLB) is a global devastating citrus disease that is mainly caused by “Candidatus Liberibacter asiaticus” (CLas). It is mostly transmitted by the insect Asian citrus psyllid (ACP, Diaphorina citri) in a persistent and proliferative manner. CLas traverses multiple barriers to complete an infection cycle and is likely involved in multiple interactions with D. citri. However, the protein–protein interactions between CLas and D. citri are largely unknown. Here, we report on a vitellogenin-like protein (Vg_VWD) in D. citri that interacts with a CLas flagellum (flaA) protein. We found that Vg_VWD was upregulated in CLas-infected D. citri. Silencing of Vg_VWD in D. citri via RNAi silencing significantly increased the CLas titer, suggesting that Vg_VWD plays an important role in the CLas–D. citri interaction. Agrobacterium-mediated transient expression assays indicated that Vg_VWD inhibits BAX- and INF1-triggered necrosis and suppresses the callose deposition induced by flaA in Nicotiana benthamiana. These findings provide new insights into the molecular interaction between CLas and D. citri.</p