An ecdysteroid-regulated 16-kDa protein homolog participates in the immune response of the crayfish Procambarus clarkii.

An ecdysteroid-regulated 16-kDa protein homolog (named Pc-E16), encoding 150 amino acid residues with a conserved MD-2-related lipid-recognition domain, was first identified in Procambarus clarkii. Phylogenetic analyses indicated similarity between Pc-E16 and 16-kDa proteins from Aplysia californica and insects. Recombinant Pc-E16 protein was successfully expressed in BL21 (DE3) Escherichia coli cells, and polyclonal antibodies against purified Pc-E16 proteins were prepared. In comparison with other tissues, Pc-E16 was highly expressed in the intestine; real-time PCR and Western blotting results indicated that Pc-E16 expression was significantly induced by lipopolysaccharides in hepatopancreas and hemocytes. Pc-E16-mediated signaling pathways were investigated by digital gene expression analysis following RNA interference targeting Pc-E16. A total of 6103 differentially expressed genes (DEGs) were identified, of which 3318 were up- and 2785 were downregulated. Many DEGs were involved in binding and catalytic activity. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis revealed that DEGs were clustered into 225 pathways, and 15 significantly enriched pathways were identified at the immune system level. In addition, the expression level of Pc-E16 in hemocytes and hepatopancreas was obviously downregulated at 48 h after dsRNA injection, and Pc-E16-RNAi treatment affected the expression levels of immune-related genes. Altogether, our results suggest that Pc-E16 is involved in the innate immune response of P. clarkii.

LncRNA ZEB2-AS1 contributes to the tumorigenesis of gastric cancer via activating the Wnt/ß-catenin pathway.

Studies have shown that long noncoding RNA Zinc finger E-box-binding homeobox 2 antisense RNA 1 (ZEB2-AS1) is involved in the progression of lung cancer, bladder cancer, and hepatocellular carcinoma. However, its role in the pathogenesis of gastric cancer remains unknown. The Wnt/ß-catenin pathway contributes to the development of gastric cancer. ZEB2-AS1 expression was firstly detected in the gastric carcinoma tissue samples as well as in gastric cancer cells. Knockdown of ZEB2-AS1 was performed by ZEB2-AS1-shRNA, and the viability, migration, invasion, and apoptosis of gastric cancer cells were determined by CCK-8, scratch assay, transwell, and flow cytometry, respectively. Furthermore, levels of Ki-67, PCNA, VEGF, MMP9, epithelial-mesenchymal transition (EMT) markers (E-cadherin, Vimentin and ZEB2), cleaved caspase 3/8/9 and PARP, active ß-catenin, c-Myc, cyclinD1, and AXIN2 were assayed by Western blot or real-time PCR. Additionally, the role and mechanism of ZEB2-AS1 were confirmed in a xenograft nude mouse model. We found ZEB2-AS1 expression was increased in gastric carcinoma samples, and it was correlated with tumor progression. Also, its expression was elevated in gastric cancer cells. Knockdown of ZEB2-AS1 reduced the proliferation, migration, invasion, and EMT, but increased the apoptosis of gastric carcinoma cells. Furthermore, ZEB2-AS1 downregulation remarkably suppressed the expression of Ki-67, PCNA, VEGF and MMP9, and the activation of Wnt/ß-catenin signaling, whereas elevated the levels of cleaved caspase 3/8/9 and PARP in gastric cancer cells. And ZEB2 overexpression reversed the effects of ZEB2-AS1 downregulation on the proliferation, EMT and inactivation of Wnt/ß-catenin signaling. Additionally, ZEB2-AS1 knockdown inhibited tumor growth, Ki-67 staining, and the expression of VEGF, MMP9, active ß-catenin, c-Myc, cyclinD1, and AXIN2 in mice. In conclusion, ZEB2-AS1 promotes the tumorigenesis of gastric carcinoma that is related to the upregulation of ZEB2 and the activation of the Wnt/ß-catenin pathway.

An Empirical Study on the Relationship between Community Sports Activities and Community Residents' Mental Health.

A full understanding of mental health can improve people's ability to identify mental diseases and cope with psychological problems, so as to improve the ability of the whole community to resist mental diseases. Community health education is particularly important in community mental health service. The traditional health education mode is carried out through lectures or paper brochures, and the effect is not significant, so we need to constantly improve the health education mode. Through the development of community mental health education and service, we can improve people's mental health quality and promote family happiness and social stability. Based on this, this study mainly analyzes the relationship between community sports activities and mental health of community residents. Physical exercise can reduce stress reaction, regulate emotion, enhance mental health, prevent, and treat mental diseases. Therefore, physical exercise has been used not only as a method to enhance physical fitness but also as an important means to regulate psychology.

A Role of Suppressor of Cytokine Signaling 2 in the Regulation of Ecdysteroid Signaling Pathway in Procambarus clarkii.

A suppressor of cytokine signaling 2 (SOCS-2) homolog was identified from the red crayfish Procambarus clarkii. Phylogenic analysis showed that P. clarkii SOCS-2 (Pc-SOCS-2) was very similar to that of invertebrates. The sequence contains a central SH2 domain, a conserved SOCS box, and a variable N-terminal domain. Real-time polymerase chain reaction showed that Pc-SOCS-2 mRNA level was higher in muscle and heart than in the other tissues examined. A 40-kDa recombinant protein was successfully expressed and purified from Escherichia coli cells as confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Pc-SOCS-2 was significantly downregulated in the hepatopancreas after 20-hydroxyecdysone injection. Knockdown of Pc-SOCS-2 gene by small interfering RNA transfection in the hepatopancreas increased the expression levels of 20-hydroxyecdysone-responsive genes at 48 hr. Taken together, these results suggest that Pc-SOCS-2 is a negative regulator of ecdysteroid signaling transduction.

Smad nuclear interacting protein, SNIP1, mediates the ecdysteroid signal transduction in red crayfish Procambarus clarkii.

A smad nuclear interacting protein 1 (SNIP1) homolog was identified in the crayfish Procambarus clarkii and this gene encoded a polypeptide of 288 amino acids. Phylogenic analysis showed that P. clarkii SNIP1 was highly homologous to that of invertebrates and shared a highly conserved FHA domain of SNIP proteins at the C-terminus. Quantitative real-time PCR (qRT-PCR) analysis showed that Pc-SNIP1 expression was higher in heart than that in other examined tissues. In addition, prokaryotic expression and purification of the recombinant SNIP1 protein were performed. SDS-PAGE and western blot analysis demonstrated that a 35 KDa recombinant protein was successfully expressed in Escherichia coli cells. The expression of Pc-SNIP1 was significantly up-regulated in hepatopancreas after 20-hydroxyecdysone induction. Knockdown of Pc-SNIP1 gene by small interfering RNA (siRNA) transfection had significant influence on the expression of some ecdysteroid-responsive genes in hepatopancreas, which were confirmed by qRT-PCR and western blot. All together, these results suggest that SNIP1 is involved in the physiological process regulated by ecdysteroid in P. clarkii.