摘要: No studies have systematically compared time-resolved and multi-tissue innate immune responses for European eels (Anguilla anguilla) with Aeromonas hydrophila infection by a high throughput method. Here, we challenged European eels with A. hydrophila (infected) or PBS (control). A low fatality rate (16.1%) was observed for the infected group at 2 d post-infection (dpi). Then we examined transcriptional profiles of intestines, livers and spleens from 12 European eels with/without infection by A. hydrophila at 6, 18 and 36 h post-infection (hpi). The results showed that the most differentially expressed genes (DEGs) were found in the spleen at 6 hpi (7569 DEGs), followed by the intestine at 6 hpi (3129 DEGs) and the liver at 18 hpi (2722 DEGs). Only 540, 41 and 130 DEGs were found in the spleen, liver, and intestine at 36 hpi, respectively. The comparison of DEG numbers and enriched KEGG pathways suggested a consistent time-course immune response in these tissues. A similar enrichment of pattern recognition receptors-related pathways including Toll-like receptors, NOD-like receptors and RIG-I-like receptors was found in the liver and spleen, but not in the intestine. Among immune-related DEGs, 62 paralogue pairs were found, and the expression trends of most paralogues were consistent. Some paralogues of immune-related DEGs had unique domains. Furthermore, large clusters representing similar tissue were shown for the intestine and spleen. A different co-expression network involved in cytokine signal transduction and inter-action existed between the liver and spleen. This study has provided time-resolved and multi-tissue transcriptome characteristics of A. anguilla in response to A. hydrophila infection.

摘要: Environmental DNA (eDNA) originates from cellular material shed by organisms into aquatic or terrestrial environments and can be sampled and monitored using metabarcoding technology, which is revolutionizing fish biodiversity monitoring. Several reviews concerning fish eDNA have focused on standard sampling methods and its applications, though a systematic review focused on marker genes, databases, and bioinformatic pipelines has not yet been published. Here, we present a comprehensive literature review of studies applying metabarcoding technology to fish eDNA for the purpose of fish biodiversity monitoring. We systematically provide the available universal primers used to amplify barcoding sequences from fish eDNA, and then discuss reference barcoding databases, relevant bioinformatic analyses, as well as developed pipelines. The performances of universal primers and their relevant reference databases are summarized. Combined use of multiple primer pairs targeted for more than one gene marker (e.g., 12S, 16S, Cytb, COI), and use of both local and public databases are recommended as approaches to improve the sensitivity and reliability of fish eDNA analyses. We also compare the effectiveness of eDNA metabarcoding to traditional approaches for monitoring fish biodiversity and highlight challenges and future perspectives associated with this new tool. Ultimately, we advocate for greater incorporation of eDNA analysis into fish biodiversity assessments to assist environmental managers.

摘要: Objectives Mammalian DNA methyltransferases are essential to re-establish global DNA methylation patterns during implantation, which is critical for transmitting epigenetic information to the next generation. In contrast, the significance of methyl-CpG binding proteins (MBPs) that bind methylated CpG remains almost unknown at this stage. We previously demonstrated that Zbtb38 (also known as CIBZ)-a zinc finger type of MBP-is required for mouse embryonic stem (ES) cell proliferation by positively regulating Nanog expression. However, the physiological function of Zbtb38 in vivo remains unclear. Materials and Methods This study used the Cre-loxP system to generate conditional Zbtb38 knockout mice. Cell proliferation and apoptosis were studied by immunofluorescence staining. Quantitative real-time PCR, immunoblotting and immunofluorescence were performed to investigate the molecular mechanisms. Results Germline loss of the Zbtb38 single allele resulted in decreased epiblast cell proliferation and increased apoptosis shortly after implantation, leading to early embryonic lethality. Heterozygous loss of Zbtb38 reduced the expression of Nanog, Sox2, and the genes responsible for epiblast proliferation, differentiation, and cell viability. Although this early lethal phenotype, Zbtb38 is dispensable for ES cell establishment and identity. Conclusions These findings indicate that Zbtb38 is essential for early embryonic development via the suppression of Nanog and Sox2 expression.

摘要: Objectives Mutant C/EBP alpha p30 (mp30), the product of C/EBP alpha double mutations (DM), lacks transactivation domain 1 and has C-terminal loss-of-function mutation. Acute myeloid leukaemia (AML) patients harbouring C/EBP alpha DM could be classified as a distinct subgroup with favourable prognosis. However, the underlying mechanism remains elusive. Materials and Methods Autophagy regulated by mp30 was detected by western blot and immunofluorescence. Immune infiltration analysis and GSEA were performed to investigate autophagic and inflammatory status of AML patients from the GSE14468 cohort. Flow cytometry was applied to analyse T cell activation. Results Mp30 inhibited autophagy by suppressing nucleus translocation of NF-kappa B. Autophagy-associated secretion of IL-1 beta was decreased in mp30-overexpressed AML cells. Bioinformatic analysis revealed that inflammatory status was attenuated, while CD8(+) T cell infiltration was upregulated in C/EBP alpha DM AML patients. Consistently, the proportion of CD8(+)CD69(+) T cells in peripheral blood mononuclear cells (PBMCs) was upregulated after co-culture with mp30 AML cell conditional culture medium. Knock-out of IL-1 beta in AML cells also enhanced CD8(+) T cell activation. Accordingly, IL-1 beta expression was significantly reduced in the bone marrow (BM) cells of C/EBP alpha DM AML patients compared to the wildtype, while the CD8(+)CD69(+) T cell proportion was specifically elevated. Conclusions C/EBP alpha DM alleviates immunosuppression of CD8(+) T cells by inhibiting the autophagy-associated secretion of IL-1 beta, which elucidated that repression of autophagy-related inflammatory response in AML patients might achieve a favourable clinical benefit.

摘要: Mesenchymal stem cells (MSCs) are heterogeneous populations with broad application prospects in cell therapy, and using specific subpopulations of MSCs can enhance their particular capability under certain conditions and achieve better therapeutic effects. However, no studies have reported how to obtain high-quality specific MSC subpopulations in vitro culture. Here, for the first time, we established a general operation process for obtaining high-quality clinical-grade cell subpopulations from human umbilical cord MSCs (hUC-MSCs) based on particular markers. We used the MSC-CD106(+) subpopulations, whose biological function has been well documented, as an example to explore and optimize the crucial links of primary preparation, pre-treatment, antibody incubation, flow sorting, quality and function test. After comprehensively evaluating the quality and function of the acquired MSC-CD106(+) subpopulations, including in vitro cell viability, apoptosis, proliferation, marker stability, adhesion ability, migration ability, tubule formation ability, immunomodulatory function and in vivo wound healing ability and proangiogenic activity, we defined an important pre-treatment scheme which might effectively improve the therapeutic efficiency of MSC-CD106(+) subpopulations in two critical clinical application scenarios-direct injection after cell sorting and post-culture injection into bodies. Based on the above, we tried to establish a general five-step operation procedure for acquiring high-quality clinical-grade MSC subpopulations based on specific markers, which cannot only improve their enrichment efficiency and the reliability of preclinical studies, but also provide valuable methodological guidance for the rapid clinical transformation of specific MSC subpopulations.