Journal: Advanced Science

Article Title: PBRM1 Deficiency in Microglia Leads to Excessive Neurogenesis and Autism‐Like Behaviors via the CCL5‐CCR1/5‐β‐catenin Signaling Axis

doi: 10.1002/advs.78086

Figure Lengend Snippet: Pbrm1 cKO microglia disrupts microglial development and homeostasis. (A) Uniform manifold approximation and projection (UMAP) visualization of 22,201 cells, including 10,727 cells from WT hippocampal samples and 11,474 cells from Pbrm1 cKO hippocampal samples. (B) UMAP visualization of microglia that were divided into 7 subclusters based on the expression of marker genes. (C) Heatmap of normalized expression levels of enriched genes in 7 microglial subclusters. (D) Single‐cell trajectories of microglia by Monocle analysis. Cells are colored by pseudotime trajectories (left panel), clusters (middle panel), or samples (right panel). (E) Bar plot showing relative proportions of each microglial subcluster between WT and cKO samples. (F) Quantitative RT‐PCR analysis showing the relative mRNA levels of P2ry12, Tmem119 , and Tnf in WT and cKO microglia. Compared to WT, cKO microglia displayed significantly lower expression of P2ry12 and Tmem119 , but higher expression of Tnf . Mean ± SEM. * p < 0.05, *** p < 0.001 by Student's t ‐test or Mann–Whitney test. n = 4 mice per group. Males (circles) and females (squares) are indicated. (G) Representative flow cytometry gating strategy for sorting IL‐1α + TNF‐α + microglia from the hippocampus of P15 WT and cKO mice. (H) Quantification of the percentage of IL‐1α + TNF‐α + microglia in the hippocampus of P15 WT and cKO mice. Mean ± SEM. *** p < 0.001 by Student's t ‐test. n = 3 mice per group. (I,J) Representative images (I) and quantifications (J) of GFAP (green) and Tuj1 (red) immunostaining of the differentiation of NSPCs that were indirectly co‐cultured with TNF‐α + , IL‐1α + , TNF‐α + IL1‐α + primary microglia in a Transwell system. In contrast to NSPCs that were indirectly co‐cultured with TNF‐α − IL‐1α − cKO microglia, TNF‐α + , IL‐1α + , TNF‐α + IL‐1α + treated NSPCs differentiated into more Tuj1 + cells and fewer GFAP + cells. Scale bar, 50 µm. Mean ± SEM. * p < 0.05, ** p < 0.01 by Student's t ‐test. n = 3 independent experiments. See also Figure .

Article Snippet: Cells were then stained with anti‐mouse TNFα FITC (130‐124‐212, Miltenyi Biotec, Germany) and anti‐mouse IL‐1α PE (130‐104‐530, Miltenyi Biotec, Germany) antibodies for phenotypic analysis.

Techniques: Expressing, Marker, Single Cell, Analysis, Quantitative RT-PCR, MANN-WHITNEY, Flow Cytometry, Immunostaining, Cell Culture

Journal: Advanced Science

Article Title: Chrysanthemum Morifolium– Derived Nanovesicles Reduce Lactate‐Associated Histone Lactylation and Modulate Microglia‐Mediated Neuroinflammation in Sepsis‐Associated Encephalopathy

doi: 10.1002/advs.78330

Figure Lengend Snippet: CRELNVs mitigated memory impairment in the hippocampus of SAE mice. (A) Schematic of the experimental procedure used to probe the effect of CRELNVs on memory impairment in the hippocampi of sepsis‐associated encephalopathy (SAE) mice. (B) Center times in open field test, n = 10. (C) Object discrimination times in novel‐object recognition test, n = 10. (D) Spontaneous alternation rates in Y‐Maze, n = 10. (E) Hematoxylin and eosin, Nissl, and TUNEL staining images. (F) Nissl staining quantitative analysis in CA3 region, n = 6. (G) TUNEL staining quantitative analysis in DG region, n = 6. (H) Interleukin‐1β (IL‐1β), (I) tumor necrosis factor‐α (TNF‐α), (J) interleukin‐6 (IL‐6), and (K) interleukin‐10 (IL‐10) levels in the hippocampi of SAE mice, n = 6. # p < 0.05, ## p < 0.01, ### p < 0.001 vs. control group; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. lipopolysaccharide (LPS) group.

Article Snippet: IL‐1β (Cat. No. KE10003), IL‐6 (Cat. No. KE10007), TNF‐α (Cat. No. KE10002), and interleukin‐10 (IL‐10; Cat. No. KE1008) Enzyme‐linked immunosorbent assay (ELISA) kits were procured from Proteintech Group (Wuhan, China).

Techniques: TUNEL Assay, Staining, Analysis, Control

Journal: Advanced Science

Article Title: Chrysanthemum Morifolium– Derived Nanovesicles Reduce Lactate‐Associated Histone Lactylation and Modulate Microglia‐Mediated Neuroinflammation in Sepsis‐Associated Encephalopathy

doi: 10.1002/advs.78330

Figure Lengend Snippet: CRELNVs influence microglial homeostasis in the hippocampus of SAE mice. (A) Representative images of immunofluorescence staining for Iba‐1in the hippocampi of SAE mice. (B) Number of Iba‐1 + cells, (C) number of total branch points/cell, and (D) total branch length for panel A, n = 6. (E) IL‐1β, (F) TNF‐α, (G) IL‐6, and (H) IL‐10 mRNA expression in the hippocampi of SAE mice determined using the quantitative polymerase chain reaction (qPCR), n = 3. # p < 0.05, ## p < 0.01, ### p < 0.001 vs. control group; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. LPS group).

Article Snippet: IL‐1β (Cat. No. KE10003), IL‐6 (Cat. No. KE10007), TNF‐α (Cat. No. KE10002), and interleukin‐10 (IL‐10; Cat. No. KE1008) Enzyme‐linked immunosorbent assay (ELISA) kits were procured from Proteintech Group (Wuhan, China).

Techniques: Immunofluorescence, Staining, Expressing, Real-time Polymerase Chain Reaction, Control

Journal: Advanced Science

Article Title: Chrysanthemum Morifolium– Derived Nanovesicles Reduce Lactate‐Associated Histone Lactylation and Modulate Microglia‐Mediated Neuroinflammation in Sepsis‐Associated Encephalopathy

doi: 10.1002/advs.78330

Figure Lengend Snippet: The neuro‐protection and Anti‐inflammation activities of CRELNVs in vitro. (A) Protective effect of CRELNVs on SH‐SY5Y cells in an inflammatory environment determined using the CCK8 assay, n = 4. (B) Microplate reader–detected NO contents in BV2 cell supernatants, n = 3. (C) IL‐1β, (D) IL‐6, (E) TNF‐α, and (F) IL‐10 levels in BV2 cell supernatants, n = 3. (G) Representative fluorescence microscopy images acquired to evaluate reactive oxygen species (ROS) content. (H, I) Flow cytometry peaks and ROS contents derived therefrom, n = 3. (J, K) Western blot analysis: NLRP3 levels in BV2 cells, n = 3. # p < 0.05, ## p < 0.01, ### p < 0.001 vs. control group; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. LPS group.

Article Snippet: IL‐1β (Cat. No. KE10003), IL‐6 (Cat. No. KE10007), TNF‐α (Cat. No. KE10002), and interleukin‐10 (IL‐10; Cat. No. KE1008) Enzyme‐linked immunosorbent assay (ELISA) kits were procured from Proteintech Group (Wuhan, China).

Techniques: In Vitro, CCK-8 Assay, Fluorescence, Microscopy, Flow Cytometry, Derivative Assay, Western Blot, Analysis, Control