Journal: International Journal of Molecular Medicine

Article Title: Remimazolam alleviates myocardial ischemia/reperfusion injury and inflammation via inhibition of the NLRP3/IL-1β pathway in mice

doi: 10.3892/ijmm.2025.5498

Figure Lengend Snippet: Rema alleviates LPS-induced release and increased expression levels of IL-1β, IL-6 and TNF-α in cultured Raw264.7 cells. (A) Experimental protocol using cultured Raw264.7 macrophages. (B) LPS induced alterations in the release of IL-1β, IL-6 and TNF-α in Raw264.7 cells, while treatment with Rema inhibited these LPS-mediated changes (n=3). (C) LPS induced alterations in the gene expression levels of IL-1β, IL-6 and TNF-α in Raw264.7 cells, while treatment with Rema inhibited these effects (n=3). (D) DAPA inhibited the LPS-induced release of IL-1β, IL-6 and TNF-α in Raw264.7 cells, while treatment with Rema did not affect the alterations induced by DAPA (n=3). *** P<0.001. DAPA, dapansutrile; LPS, lipopolysaccharide; ns, not significant; Rema, remimazolam; RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: Cells were cultured in DMEM without FBS and pre-treated with 100 μ g/ml Rema (diluted in DMEM) for 20 min, and subsequently treated with lipopolysaccharide (LPS; 0.5 μ g/ml diluted in DMEM; cat. no. HY-D1056H, MedChemExpress) for 24 h in incubator at 37°C.

Techniques: Expressing, Cell Culture, Gene Expression, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Frontiers in Microbiology

Article Title: Transcriptomic Analysis Reveals Competitive Growth Advantage of Non-pigmented Serratia marcescens Mutants

doi: 10.3389/fmicb.2021.793202

Figure Lengend Snippet: Mutation trend of Serratia marcescens during continuous passages. (A) Colonies on Luria-Bertani (LB) plate at the representative generations of passage. The percentage of color mutants is labeled upside. (B) The percentage of wild-type strains and mutants in each generation of passage.

Article Snippet: In this study, the wild-type S. marcescens strain SCQ1 (CCTCC AB 2010221) was isolated from the hemolymph of diseased silkworm larva ( Bombyx mori ) previously, which was stored at −80°C ( ).

Techniques: Mutagenesis, Labeling

Journal: Frontiers in Microbiology

Article Title: Transcriptomic Analysis Reveals Competitive Growth Advantage of Non-pigmented Serratia marcescens Mutants

doi: 10.3389/fmicb.2021.793202

Figure Lengend Snippet: The phenotypic patterns of wild-type S. marcescens and its spontaneous mutants. (A) The colony of the wild-type strain SCQ1 and the four mutants on LB plate. (B) Growth of different S. marcescens strains in LB medium. The OD 600 values are monitored every 2 h for 100 h using the Bioscreen C instrument with three technical replicates. Different lines and symbols represent different bacterial strains. The data are expressed as mean ± SD. Several time points are labeled and the details are shown in (C) . (D) The relative prodigiosin production in different S. marcescens strains is measured after 16 h incubation at 28°C. The values are calculated and expressed as mean ± SD ( n = 3, * p < 0.05, ** p < 0.01).

Article Snippet: In this study, the wild-type S. marcescens strain SCQ1 (CCTCC AB 2010221) was isolated from the hemolymph of diseased silkworm larva ( Bombyx mori ) previously, which was stored at −80°C ( ).

Techniques: Labeling, Incubation

Journal: Frontiers in Microbiology

Article Title: Transcriptomic Analysis Reveals Competitive Growth Advantage of Non-pigmented Serratia marcescens Mutants

doi: 10.3389/fmicb.2021.793202

Figure Lengend Snippet: The mRNA expression levels of pigA-pigN in the wild-type strain SCQ1 and the four spontaneous mutants by quantitative real-time RT-PCR (qRT-PCR) analysis. Relative mRNA levels are quantified using rpoB as internal control and the data are displayed as mean ± SE ( n = 3, * p < 0.05, ** p < 0.01).

Article Snippet: In this study, the wild-type S. marcescens strain SCQ1 (CCTCC AB 2010221) was isolated from the hemolymph of diseased silkworm larva ( Bombyx mori ) previously, which was stored at −80°C ( ).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Frontiers in Microbiology

Article Title: Transcriptomic Analysis Reveals Competitive Growth Advantage of Non-pigmented Serratia marcescens Mutants

doi: 10.3389/fmicb.2021.793202

Figure Lengend Snippet: Summary of transcriptome sequencing for SCQ1 and SCQ1-3M.

Article Snippet: In this study, the wild-type S. marcescens strain SCQ1 (CCTCC AB 2010221) was isolated from the hemolymph of diseased silkworm larva ( Bombyx mori ) previously, which was stored at −80°C ( ).

Techniques: Sequencing

Journal: Frontiers in Microbiology

Article Title: Transcriptomic Analysis Reveals Competitive Growth Advantage of Non-pigmented Serratia marcescens Mutants

doi: 10.3389/fmicb.2021.793202

Figure Lengend Snippet: Transcriptomic analysis of the wild-type strain SCQ1 and the non-pigmented spontaneous mutant SCQ1-3M. (A) The volcano plot shows a general gene expression pattern of the SCQ1-3M. The red, green, and black dots represent significant upregulated, significant downregulated and, non-significant genes, respectively. The |log 2 (FC)| ≥ 1; FDR < 0.05 is set as the threshold for significantly DEGs. (B) Gene Ontology (GO) annotations of DEGs of the mutant SCQ1-3M. The DEGs are grouped into three main categories which are “cellular component,” “molecular function,” and “biological process” according to the GO data library. The red and green bars represent significant upregulated and downregulated DEGs, respectively. The Y-axis indicates the number of DEGs in each category. (C) The top 20 enriched KEGG pathways of DEGs. X- and Y-axes represent the Rich factor and the KEGG pathways, respectively.

Article Snippet: In this study, the wild-type S. marcescens strain SCQ1 (CCTCC AB 2010221) was isolated from the hemolymph of diseased silkworm larva ( Bombyx mori ) previously, which was stored at −80°C ( ).

Techniques: Mutagenesis, Gene Expression

Journal: Frontiers in Microbiology

Article Title: Transcriptomic Analysis Reveals Competitive Growth Advantage of Non-pigmented Serratia marcescens Mutants

doi: 10.3389/fmicb.2021.793202

Figure Lengend Snippet: The characteristics of the Δ slyA mutant. (A) The mRNA expression level of slyA in different spontaneous mutants. (B) Colonies of the mutant Δ slyA , the control strains Δ slyA -pJQ200SK and SCQ1-pJQ200SK, and the complemented strain Δ slyA -pJQ200SK- slyA on LB plate inoculated at 28°C for 36 h. (C) The relative prodigiosin production levels in the strain SCQ1, Δ slyA , Δ slyA -pJQ200SK, Δ slyA -pJQ200SK- slyA , and SCQ1-pJQ200SK are measured after 16 h incubation at 28°C. The A 534 /OD 600 values are calculated and expressed as mean ± SD. (D) The mRNA expression level of slyA in the recombinant strains. (E) The growth curves of the strain SCQ1, Δ slyA , and Δ slyA -pJQ200SK- slyA . The OD 600 values are monitored every 2 h for 100 h using the Bioscreen C instrument with three technical replicates. Different lines and symbols represent different bacterial strains. The data are expressed as mean ± SD. Several time points are labeled and the details are shown in (F) . (G) The mRNA expression levels of pigA-pigN in the wild-type strain SCQ1 and the mutant Δ slyA . (H) The mRNA expression levels of pigA-pigN in Δ slyA -pJQ200SK- slyA and the control strains with an empty plasmid pJQ200SK. (I) Symptom of silkworm larvae infected by the strain SCQ1 and Δ slyA . The mock group represents silkworm larvae injected with sterile saline as the negative control. (J) Cumulative survival of silkworm larvae injected with SCQ1 [100 colony-forming units (CFUs) per larva], Δ slyA (100 CFUs per larva), and sterile saline. (K) The LD 50 values of the strain SCQ1 and Δ slyA in the silkworm larvae. (L) The LT 50 of the strain SCQ1 and Δ slyA in the silkworm larvae. The data were expressed as mean ± SD ( n = 3, * p < 0.05, ** p < 0.01).

Article Snippet: In this study, the wild-type S. marcescens strain SCQ1 (CCTCC AB 2010221) was isolated from the hemolymph of diseased silkworm larva ( Bombyx mori ) previously, which was stored at −80°C ( ).

Techniques: Mutagenesis, Expressing, Control, Incubation, Recombinant, Labeling, Plasmid Preparation, Infection, Injection, Sterility, Saline, Negative Control

Journal: Frontiers in Microbiology

Article Title: Transcriptomic Analysis Reveals Competitive Growth Advantage of Non-pigmented Serratia marcescens Mutants

doi: 10.3389/fmicb.2021.793202

Figure Lengend Snippet: The components of the T6SS detected in the present study. All the labeled genes are significantly downregulated in the spontaneous non-pigmented mutant SCQ1-3M. The DEGs of T6SS related secretion proteins in the present study were listed at lower-left. OM, outer membrane; IM, inner membrane.

Article Snippet: In this study, the wild-type S. marcescens strain SCQ1 (CCTCC AB 2010221) was isolated from the hemolymph of diseased silkworm larva ( Bombyx mori ) previously, which was stored at −80°C ( ).

Techniques: Labeling, Mutagenesis, Membrane

Journal: Frontiers in Microbiology

Article Title: Transcriptomic Analysis Reveals Competitive Growth Advantage of Non-pigmented Serratia marcescens Mutants

doi: 10.3389/fmicb.2021.793202

Figure Lengend Snippet: The potential mechanisms contribute to competitive growth advantage in S. marcescens spontaneous mutants under laboratory conditions. Both the wild-type strain and its spontaneous mutants preserve the complete prodigiosin synthesis gene cluster ( Pig gene cluster). In the pigmented wild-type strain (left side), the pig genes are highly expressed under the control of transcriptional regulators, and the pigment is massively produced. The abundant prodigiosin is stored in membrane vesicles and secreted outside the cell by vesicular transport pathways to maintain lower intracellular concentrations which avoid feedback inhibition. The wild-type strain also consumes necessary resources to maintain some high-cost systems, e.g., T6SS. The amino acids required for prodigiosin synthesis and other systems are imported via amino acid transport systems, in which only a small amount of amino acids is degraded. In the non-pigmented spontaneous mutant (right side), the expression of the pig genes is highly suppressed by several factors. The transcriptional regulators, e.g., the positive regulator slyA , are down-expressed, which results in downregulated pig genes. Lack of essential membrane proteins for vesicle formation also potentially contribute to prodigiosin dyssynthesis. In this scenario, prodigiosin is neither correctly stored nor secreted outside the cells, which leads to strong feedback inhibition effects. Besides the pigment synthesis system, the non-pigmented mutant also shuts down T6SS or other high-cost systems to save resources. In addition, both amino acid transport- and degradation-pathways associated proteins are over-expressed in the non-pigmented mutant, which supplies extra nutrients and energies for bacterial growth. Therefore, the non-pigmented mutant shows a competitive growth advantage over the wild-type strain, which rapidly replaces the latter in laboratory conditions.

Article Snippet: In this study, the wild-type S. marcescens strain SCQ1 (CCTCC AB 2010221) was isolated from the hemolymph of diseased silkworm larva ( Bombyx mori ) previously, which was stored at −80°C ( ).

Techniques: Control, Produced, Membrane, Inhibition, Mutagenesis, Expressing