9 research outputs found

    Image2_The new ceRNA crosstalk between mRNAs and miRNAs in intervertebral disc degeneration.TIFF

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    Degeneration of the intervertebral disc has been linked to lower back pain. To date, pathophysiological mechanisms of intervertebral disc degeneration (IDD) remain unclear; it is meaningful to find effective diagnostic biomarkers and new therapeutic strategies for IDD. This study aimed to reveal the molecular mechanism of IDD pathogenesis from the multidimensional transcriptomics perspective. Here, we acquired IDD bulk omics datasets (GSE67567 and GSE167199) including mRNA, microRNA expression profiles, and single-cell RNA sequencing (GSE199866) from the public Gene Expression Omnibus (GEO) database. Through principal component analysis and Venn analysis, we found different expression patterns in the IDD transcription level and identified 156 common DEGs in both bulk datasets. GO and KEGG functional analyses showed these dysregulators were mostly enriched in the collagen-containing extracellular matrix, cartilage development, chondrocyte differentiation, and immune response pathways. We also constructed a potentially dysregulated competing endogenous RNA (ceRNA) network between mRNAs and miRNAs related to IDD based on microRNA target information and co-expression analysis of RNA profiles and identified 36 ceRNA axes including ZFP36/miR-155-5p/FOS, BTG2/hsa-miR-185-5p/SOCS3, and COL9A2/hsa-miR-664a-5p/IBA57. Finally, in integrating bulk and single-cell transcriptome data analyses, a total of three marker genes, COL2A1, PAX1, and ZFP36L2, were identified. In conclusion, the key genes and the new ceRNA crosstalk we identified in intervertebral disc degeneration may provide new targets for the treatment of IDD.</p

    Table1_The new ceRNA crosstalk between mRNAs and miRNAs in intervertebral disc degeneration.XLS

    No full text
    Degeneration of the intervertebral disc has been linked to lower back pain. To date, pathophysiological mechanisms of intervertebral disc degeneration (IDD) remain unclear; it is meaningful to find effective diagnostic biomarkers and new therapeutic strategies for IDD. This study aimed to reveal the molecular mechanism of IDD pathogenesis from the multidimensional transcriptomics perspective. Here, we acquired IDD bulk omics datasets (GSE67567 and GSE167199) including mRNA, microRNA expression profiles, and single-cell RNA sequencing (GSE199866) from the public Gene Expression Omnibus (GEO) database. Through principal component analysis and Venn analysis, we found different expression patterns in the IDD transcription level and identified 156 common DEGs in both bulk datasets. GO and KEGG functional analyses showed these dysregulators were mostly enriched in the collagen-containing extracellular matrix, cartilage development, chondrocyte differentiation, and immune response pathways. We also constructed a potentially dysregulated competing endogenous RNA (ceRNA) network between mRNAs and miRNAs related to IDD based on microRNA target information and co-expression analysis of RNA profiles and identified 36 ceRNA axes including ZFP36/miR-155-5p/FOS, BTG2/hsa-miR-185-5p/SOCS3, and COL9A2/hsa-miR-664a-5p/IBA57. Finally, in integrating bulk and single-cell transcriptome data analyses, a total of three marker genes, COL2A1, PAX1, and ZFP36L2, were identified. In conclusion, the key genes and the new ceRNA crosstalk we identified in intervertebral disc degeneration may provide new targets for the treatment of IDD.</p

    Western blot analysis of C3 and Bf in zebrafish egg cytosol (lane M: molecular marker; lane H: human serum; lane Z: zebrafish egg cytosol).

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    <p>The egg cytosol was electrophoresed on 12% SDS-PAGE gel using the buffer system of Laemmli. The proteins separated were blotted on nitrocellulose membrane and immunostained with rabbit anti-human C3 antibody or goat anti-human Bf antibody, followed by staining with HRP-labeled anti-rabbit IgG and HRP-labeled anti-goat IgG, respectively.</p

    Effects of anti-C3 antibody and heating on the bacteriolytic activity of zebrafish egg cytosol.

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    <p>The egg cytosol was pre-incubated with anti-C3 antibody at different concentrations (A), or inactivated by heating at 45°C (B), and then mixed with <i>E. coli</i> suspension. After incubation at 25°C for 2 h, the bacteriolytic activities were measured by colony forming unit assay. * means <i>p</i><0.05.</p

    Influences of antibodies and chemical inhibitors on the bateriolytic activity of zebrafish egg cytosol.

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    <p>The egg cytosol was pre-incubated with complement component antibodies and chemicals at optimum concentrations, and mixed with <i>E. coli</i> suspension. After incubation at 25°C for 2 h, the bacteriolytic activities were measured by colony forming unit assay.</p

    Image1_The new ceRNA crosstalk between mRNAs and miRNAs in intervertebral disc degeneration.TIFF

    No full text
    Degeneration of the intervertebral disc has been linked to lower back pain. To date, pathophysiological mechanisms of intervertebral disc degeneration (IDD) remain unclear; it is meaningful to find effective diagnostic biomarkers and new therapeutic strategies for IDD. This study aimed to reveal the molecular mechanism of IDD pathogenesis from the multidimensional transcriptomics perspective. Here, we acquired IDD bulk omics datasets (GSE67567 and GSE167199) including mRNA, microRNA expression profiles, and single-cell RNA sequencing (GSE199866) from the public Gene Expression Omnibus (GEO) database. Through principal component analysis and Venn analysis, we found different expression patterns in the IDD transcription level and identified 156 common DEGs in both bulk datasets. GO and KEGG functional analyses showed these dysregulators were mostly enriched in the collagen-containing extracellular matrix, cartilage development, chondrocyte differentiation, and immune response pathways. We also constructed a potentially dysregulated competing endogenous RNA (ceRNA) network between mRNAs and miRNAs related to IDD based on microRNA target information and co-expression analysis of RNA profiles and identified 36 ceRNA axes including ZFP36/miR-155-5p/FOS, BTG2/hsa-miR-185-5p/SOCS3, and COL9A2/hsa-miR-664a-5p/IBA57. Finally, in integrating bulk and single-cell transcriptome data analyses, a total of three marker genes, COL2A1, PAX1, and ZFP36L2, were identified. In conclusion, the key genes and the new ceRNA crosstalk we identified in intervertebral disc degeneration may provide new targets for the treatment of IDD.</p

    Effects of divalent cation chelators EGTA and EGTA on the bacteriolytic activity of zebrafish egg cytosol.

    No full text
    <p>The egg cytosol was pre-incubated with EGTA, EDTA, EDTA with Mg<sup>2+</sup> or EDTA with Ca<sup>2+</sup> at optimum concentrations, and then mixed with <i>E. coli</i> suspension. After incubation at 25°C for 2 h, the bacteriolytic activities were measured by colony forming unit assay.</p

    Characteristics of bacteriolytic activity in zebrafish egg cytosol.

    No full text
    <p>The egg cytosol filtered through 0.22 µm filter was mixed with <i>E. coli</i> suspension and incubated at 25°C for different periods (A) or at different temperature for 2 h (B). The bacteriolytic activities were determined by colony forming unit assay.</p
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