Cytoplasmic tail of transmembrane dscam controls antibacterial responses by regulating cell proliferation-related genes in hemocytes of Chinese mitten crab (Eriocheir sinensis).

In arthropods, the involvement of Dscam (Down syndrome cell adhesion molecule) in innate immunity has been extensively demonstrated. Its cytoplasmic tail contains multiple conserved functional sites, which indicates its involvement in different intracellular signaling pathways. In this study, we focused on the role of the cytoplasmic tail of Dscam in the Chinese mitten crab (Eriocheir sinensis) immune defense. In the group with cytoplasmic tail knockdown (the site was located on constant exons 37 and 38), 3885 differentially expressed genes (DEGs) were identified. The DEGs were enriched in small molecule binding, protein-containing complex binding, and immunity-related pathways. The expression of selected genes were validated using quantitative real-time reverse transcription PCR. We identified key Cell cycle, Janus kinase (JAK)-signal transducer, activator of transcription (STAT) and mitogen-activated protein kinase (MAPK) signaling pathway genes, the results indicated that the cytoplasmic tail of Dscam controls antibacterial responses by regulating cell proliferation-related genes in hemocytes.

Research Progress and the Prospect of Damping Magnesium Alloys.

As the lightest structural metal material, magnesium alloys possess good casting properties, high electrical and thermal conductivity, high electromagnetic shielding, and excellent damping properties. With the increasing demand for lightweight, high-strength, and high-damping structural materials in aviation, automobiles, rail transit, and other industries with serious vibration and noise, damping magnesium alloy materials are becoming one of the important development directions of magnesium alloys. A comprehensive review of the progress in this field is conducive to the development of damping magnesium alloys. This review not only looks back on the traditional damping magnesium alloys represented by Mg-Zr alloys, Mg-Cu-Mn alloys, etc. but also introduces the new damping magnesium materials, such as magnesium matrix composites and porous magnesium. But up to now, there have still been some problems in the research of damping magnesium materials. The effect of spiral dislocation on damping is still unknown and needs to be studied; the contradiction between damping performance and mechanical properties still lacks a good balance method. In the future, the introduction of more diversified damping regulating methods, such as adding other elements and reinforcements, optimizing the manufacturing method of damping magnesium alloy, etc., to solve these issues, will be the development trend of damping magnesium materials.

CD276 promotes epithelial-mesenchymal transition in esophageal squamous cell carcinoma through the TGF-ß/SMAD signaling.

OBJECTIVE: Aberrant expression of CD276 has been reported in malignant tumors. However, the exact role and mechanisms of CD276 influence the progression of esophageal squamous cell carcinoma (ESCC) still need to be understood. METHODS: Bioinformatics analysis of data from The Cancer Genome Atlas and Gene Expression Omnibus databases, along with immunohistochemistry staining, was used to explore the expression patterns of CD276 in ESCC. Cell counting kit-8 and Transwell assays were employed to evaluate the effects of CD276 expression on tumor cell proliferation and motility. Western blotting and Transwell assays were used to explore the potential pathways through which CD276 mediates the progression of ESCC. Moreover, the in vivo role of CD276 in tumor progression was investigated by establishing a lung metastasis mouse model. RESULTS: A significant upregulation of CD276 was observed in ESCC tissues compared to adjacent tissues. The inhibition of CD276 had no evident impact on ESCC cell proliferation but notably hindered their migratory and invasive properties and the expression of epithelial-mesenchymal transition (EMT) markers. Inversely, overexpressing CD276 led to an upregulation of EMT markers, underscoring the capacity of CD276 to amplify the motility of ESCC cells. Furthermore, CD276 was found to enhance the migratory and invasive abilities of ESCC cells by activating the TGF-ß/SMAD signaling but not the PI3K/AKT pathway. In vivo studies demonstrated that CD276 facilitates pulmonary metastasis. CONCLUSION: CD276 is significant upregulation in ESCC tissues and facilitates the EMT process in ESCC cells via the TGF-ß/SMAD signaling, thus promoting the progression of ESCC.

Exposure to nanopolystyrene and phoxim at ambient concentrations causes oxidative stress and inflammation in the intestines of the Chinese mitten crab (Eriocheir sinensis).

Nanopolystyrene (NP) and phoxim (PHO) are common environmental pollutants in aquatic systems. We evaluated the toxic effects of exposure to ambient concentrations of NP and/or PHO in the intestines of the Chinese mitten crab (Eriocheir sinensis). Our study showed that histopathological changes were observed in the intestines. Specifically, NP and/or PHO exposure increased intraepithelial lymphocytes. Furthermore, NP and/or PHO exposure induced oxidative stress, as evidenced by a significant decrease in superoxide dismutase activity (SOD), peroxidase activity (POD), and total antioxidant capacity (T-AOC). Pro-inflammatory gene expression and transcriptome analysis demonstrated that NP and/or PHO exposure induced the intestinal inflammatory response. Transcriptome results showed that NP and/or PHO exposure upregulated the NF-κB signaling pathway, which is considered a key pathway in the inflammatory response. Additionally, the expression of pro-inflammatory genes significantly increased after a single exposure to NP or PHO, but it exhibited a significant decrease after the co-exposure. The downregulation of these genes in the co-exposure group likely suggested that the co-exposure mitigated intestinal inflammation response in E. sinensis. Collectively, our findings mainly showed that NP and/or PHO exposure at ambient concentrations induces oxidative stress and inflammatory response in the intestines of E. sinensis.

Toxic effects of nanopolystyrene and cadmium on the intestinal tract of the Chinese mitten crab (Eriocheir sinensis).

Nanopolystyrene (NP) and cadmium (Cd) are ubiquitous contaminants in aquatic systems. The present study aimed to investigate the toxic effects of exposure to ambient concentrations of NP and/or Cd on the intestinal tract of the Chinese mitten crab (Eriocheir sinensis). Exposure to NP and/or Cd induced oxidative stress, as evidenced by a significant increase in lipid peroxide content (LPO), total antioxidant capacity (T-AOC), and peroxidase activity (POD), and significant decreases in superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) activities in E. sinensis. In addition, exposure to NP and/or Cd imbalanced the homeostasis of the intestinal microbiota, as demonstrated by the significantly increased abundance of Spiroplasma. Transcriptomic and metabolomic analyses were performed to investigate the mechanisms underlying intestinal toxicity. Our results showed that ferroptosis, ABC transporters, phosphotransferase system, apoptosis, and leukocyte transendothelial migration were disturbed after exposure to NP and/or Cd. In particular, Cd exposure affected mucin type O-glycan biosynthesis, purine metabolism, and neuroactive ligand-receptor interaction. Intriguingly, co-exposure to NP and Cd might mitigate intestinal toxicity by decreasing oxidative stress and affecting these pathways. Taken together, our study clearly demonstrates that exposure to NP and/or Cd at environmentally relevant concentrations causes intestinal toxicity in E. sinensis.

Research Progress on the Damping Mechanism of Magnesium Alloys.

Magnesium alloys with high damping, high specific strength and low density have attracted great attention in recent years. However, the application of magnesium alloys is limited by the balance between their mechanical and damping properties. The strength and plasticity of magnesium alloys with high damping performance often cannot meet the industrial requirements. Understanding the damping mechanism of magnesium alloys is significant for developing new materials with high damping and mechanical properties. In this paper, the damping mechanisms and internal factors of the damping properties of magnesium alloys are comprehensively reviewed. Some damping mechanisms have been studied by many scholars, and it has been found that they can be used to explain damping performance. Among existing damping mechanisms, the G-L dislocation theory, twin damping mechanism and interface damping mechanism are considered common. In addition, some specific long-period stacking ordered (LPSO) phases' crystal structures are conducive to dislocation movement, which is good for improving damping performance. Usually, the damping properties of magnesium alloys are affected by some internal factors directly, such as dislocation density, solute atoms, grain texture and boundaries, etc. These internal factors affect damping performance by influencing the dissipation of energy within the crystal. Scholars are working to find novel damping mechanisms and suitable solute atoms that can improve damping performance. It is important to understand the main damping mechanisms and the internal factors for guiding the development of novel high-damping magnesium alloys.

Alternatively spliced exon 33 in Dscam controls antibacterial responses through regulating cellular endocytosis and regulation of actin cytoskeleton gene expression in the hemocytes of the Chinese mitten crab (Eriocheir sinensis).

It has been widely established that Down syndrome cell adhesion molecule (Dscam) regulates arthropod cellular endocytosis. However, the signal transduction pathways and molecular mechanisms of the regulatory process remain unclear. Our previous study identified a Dscam-mediated immune signal transduction pathway that regulates cellular antimicrobial peptide expression, and a conserved endocytosis motif encoded by exon 33 in the cytoplasmic tail of transmembrane Dscam. Therefore, the present study aimed to determine the transcriptional response of the Chinese mitten crab (Eriocheir sinensis) Dscam with a cytoplasmic tail encoded by different exons. In the group of exon 32 knockdown, 306 differentially expressed genes (DEGs) were identified, and 3579 differentially expressed genes (DEGs) were identified in the group of exon 33 knockdown (green fluorescent protein, (GFP) as control). The DEGs were enriched in small molecule binding, protein-containing complex binding, and immunity-related pathways. Quantitative real-time reverse transcription PCR validated the data for selected genes. Our study contributes to the understanding of the immune defense mechanism in E. sinensis and the development of the innate immune system, thus providing insights into disease control and prevention in aquaculture.

Effects of Solution Treatment on Damping Capacities of Binary Mg-X (X = Ga and Er) Alloys.

Designing new materials for vibration and noise reduction that are lightweight is of great significance for industrial development. Magnesium (Mg) alloy is considered one of the best damping metal structural materials because of its low density, high specific strength, good energy storage characteristics and rich resources. Solution atoms have an important effect on the damping capacities of Mg alloys, but the relevant laws have not been completely clarified. In this work, two kinds of alloying elements (Ga and Er) with various atomic sizes were selected to study the metallographic structure and damping capacities of binary Mg-X (X = Ga and Er) alloys in the as-cast and solid solution states, respectively. Solution treatment can improve the damping capacities of binary Mg-X (X = Ga and Er) alloys, and the damping mechanisms of the two solid solution alloys are consistent with the G-L damping mechanism. The influence of alloy elements with different atomic sizes on damping capacities is also different. This influence is due to the various radii of solute atoms and Mg atoms which can result in different degrees of lattice distortion. This work provides a research basis for development and design of high-performance damping Mg alloy materials.

Aspartate Alleviates Colonic Epithelial Damage by Regulating Intestinal Stem Cell Proliferation and Differentiation via Mitochondrial Dynamics.

SCOPE: Proliferation and differentiation of intestinal stem cells (ISCs) are crucial for functional restoration after injury, which can be regulated by nutritional molecules. Aspartate is implicated in maintaining intestinal barrier after injury, but underlying mechanisms remain elusive. Here, this study seeks to investigate if aspartate alleviates colonic epithelial damage by regulating ISC function, and to elucidate its mechanisms. METHODS AND RESULTS: Eight-week-old male C57BL/6 mice supplement with or without 1% L-aspartate are subjected to drinking water or 2.5% DSS to induce colitis. In this study, aspartate administration alleviates the severity of colitis, as indicated by reduced body weight loss, colon shortening, and inhibited pro-inflammatory cytokine expression in DSS-challenged mice. Additionally, aspartate promotes colonic epithelial cell proliferation and differentiation after DSS-induced damage in mice. Pretreatment with aspartate not only enhances ISC proliferation but also induces ISC differentiation toward enterocytes and goblet cells, which prevent TNF-α-induced colonoid damage. Mechanistically, aspartate ameliorates DSS/TNF-α-induced perturbation of mitochondrial metabolism and maintains mitochondrial dynamics in colonic epithelium and colonoids. Moreover, aspartate-mediated ISC proliferation and differentiation are primarily dependent on mitochondrial fusion rather than fission. CONCLUSIONS: The findings indicate that aspartate promotes ISC proliferation and differentiation to alleviate colonic epithelial damage by regulation of mitochondrial metabolism and dynamics.

Kinetics of changes in gene and microRNA expression related with muscle inflammation and protein degradation following LPS-challenge in weaned piglets.

To test the dynamic changes of the expression of genes and microRNA in the gastrocnemius muscle after LPS challenge, 36 piglets were assigned to a control group (slaughtered 0 h after saline injection) and LPS groups (slaughtered at 1 h, 2 h, 4 h, 8 h, and 12 h after LPS treatment, respectively). After LPS treatment, the mRNA expression of IL-1ß, IL-6, and TNF-α reached maximal levels at 1 h, 2 h, and 1 h, respectively (P < 0.05), and mRNA expression of TLR4, NODs, muscle-specific ring finger 1, and muscle atrophy F-box peaked at 12 h (P < 0.05). Moreover, the expression of miR-122, miR-135a, and miR-370 reduced at 1 h, 1 h, and 2 h, respectively (P < 0.05), and miR-34a, miR-224, miR-132, and miR-145 reached maximum expression levels at 1 h, 1 h, 2 h, and 4 h, respectively (P < 0.05). These results suggested that mRNA expression of pro-inflammatory cytokines was elevated in the early stage, mRNA expression of genes related to TLR4 and NODs signaling pathways and protein degradation increased in the later phase, and the expression of microRNA related to muscle inflammation and protein degradation changed in the early stage after LPS injection.

Effect of flaxseed oil on muscle protein loss and carbohydrate oxidation impairment in a pig model after lipopolysaccharide challenge.

Flaxseed oil is rich in α-linolenic acid (ALA), which is the metabolic precursor of EPA and DHA. The present study investigated the effect of flaxseed oil supplementation on lipopolysaccharide (LPS)-induced muscle atrophy and carbohydrate oxidation impairment in a piglet model. Twenty-four weaned pigs were used in a 2 × 2 factorial experiment including dietary treatment (5 % maize oil v. 5 % flaxseed oil) and LPS challenge (saline v. LPS). On day 21 of treatment, the pigs were injected intraperitoneally with 100 µg/kg body weight LPS or sterile saline. At 4 h after injection, blood, gastrocnemius muscle and longissimus dorsi muscle were collected. Flaxseed oil supplementation increased ALA, EPA, total n-3 PUFA contents, protein:DNA ratio and pyruvate dehydrogenase complex quantity in muscles (P < 0·05). In addition, flaxseed oil reduced mRNA expression of toll-like receptor (TLR) 4 and nucleotide-binding oligomerisation domain protein (NOD) 2 and their downstream signalling molecules in muscles and decreased plasma concentrations of TNF-α, IL-6 and IL-8, and mRNA expression of TNF-α, IL-1ß and IL-6 (P < 0·05). Moreover, flaxseed oil inclusion increased the ratios of phosphorylated protein kinase B (Akt) 1:total Akt1 and phosphorylated Forkhead box O (FOXO) 1:total FOXO1 and reduced mRNA expression of FOXO1, muscle RING finger (MuRF) 1 and pyruvate dehydrogenase kinase 4 in muscles (P < 0·05). These results suggest that flaxseed oil might have a positive effect on alleviating muscle protein loss and carbohydrates oxidation impairment induced by LPS challenge through regulation of the TLR4/NOD and Akt/FOXO signalling pathways.

Alternatively spliced down syndrome cell adhesion molecule (Dscam) controls innate immunity in crab.

Alternatively-spliced hypervariable immunoglobulin domain-encoding molecules, called Down syndrome cell adhesion molecule (Dscam), have been widely detected as components of the arthropod immune system. Although its ability to specifically bind pathogens and enable phagocytosis of bacteria has been elucidated, the signal transduction mechanisms or effectors that activate post-Dscam-binding pathogens remain poorly characterized. Here, we reveal the alternative splicing exons of Dscam's cytoplasmic tail and its isoforms in the hemocytes of crab (Eriocheir sinensis), showing that the expression of Dscam was acutely induced after an immune challenge, which suggested its functioning for innate immunity. Significantly decreased expression levels of antimicrobial molecular peptides (AMPs) were detected in Dscam-silenced crab hemocytes in vitro, which coincided with their vulnerability to infection by Staphylococcus aureus and higher bacterial concentrations occurring in Dscam-silenced crabs in vivo Further experimental investigation demonstrated that Dscam-regulated AMP expression via the Src homology (SH)3-binding domain in the first constant exon translated protein of the cytoplasmic tail bound with the SH3 domain of the Dock, an SH3/SH2 adaptor protein required for axon guidance. Dock promoted extracellular signal-regulated kinase (ERK) phosphorylation via indirect binding and then regulated dorsal phosphorylation and translocation from the cytoplasm to the nucleus, subsequently promoting AMP expression for the effective removal of bacteria. To the best of our knowledge, this comprehensive study is the first to highlight the critical role of the alternatively-spliced Dscam cytoplasmic tail in antimicrobial control activity. It also suggests possible cross-talk occurring between Dscam and other pattern recognition receptors.

B52 promotes alternative splicing of Dscam in Chinese mitten crab, Eriocheir sinensis.

B52 is a member of the classical serine/arginine (SR)-rich proteins, which are phylogenetically conserved and play significant roles in mRNA maturation, including alternative splicing. In the present study, the docking site, selector sequences and locus control region of the Chinese mitten crab (Eriocheir sinensis) Down syndrome cell adhesion molecule (EsDscam) were identified. Alternative splicing of Dscam is essential to generate different isoforms. We also isolated and characterised the B52 gene from E. sinensis (EsB52). The 876 bp open reading frame of EsB52 encodes a 291 amino acid residue polypeptide, and EsB52 has two RNA recognition motifs (RRMs) at the N-terminus and an arginine/serine-rich domain at the C-terminus. Each RRM contains two degenerate short submotifs, RNP-1 and RNP2. Analysis of tissue distribution revealed that EsB52 mRNA expression was widespread in all tested tissues, and especially high in brain and hemocytes. In hemocytes, EsB52 was upregulated significantly after stimulation with pathogen-associated molecular patterns and bacteria. Furthermore, EsB52 RNAi decreased the number of Ig7 inclusion in mRNA rather than Ig2 or Ig3. Taken together, these findings suggest that EsB52 acts as an alternative splicing activator of EsDscam.

JAK/STAT signalling regulates antimicrobial activities in Eriocheir sinensis.

The Janus kinase/signal transducers and activators of transcription (JAK/STAT) signalling pathway plays a significant role in immune responses to pathogens. In invertebrates, three core components (Domeless, Hopscotch and STAT92E) of the JAK/STAT pathway were first identified in Drosophila melanogaster. In the present study, we report the cloning and characterisation of DOME, JAK and STAT from Eriocheir sinensis. EsDOME, EsJAK and EsSTAT are 1299, 1110 and 794 amino acid proteins encoded by 4200, 3333 and 2385 bp open reading frames, respectively. Bioinformatics analysis revealed that all three share specific domains with corresponding functions. Quantitative real-time PCR showed that all three components were highly expressed in various tissues including the gill, hepatopancreas and hemocytes. Moreover, EsDOME, EsJAK and EsSTAT were upregulated significantly in hemocytes after bacterial challenge. In addition, the JAK/STAT signalling pathway positively regulates antimicrobial peptide expression during the antibacterial immune response. Thus, the JAK/STAT pathway plays a critical role in bacterial immune responses in E. sinensis.