Journal: bioRxiv

Article Title: Midazolam suppresses glioma progression by attenuating neuronal activity and downregulating IGF1 signaling

doi: 10.64898/2026.03.31.715727

Figure Lengend Snippet: (A) Differentially expressed genes and overall expression trends of cortical neurons from seven different groups. “D vs A”: log₂FC of KCls vs Control, “G vs D”: log₂FC of MDZs vs KCls. (B) Gene expression profiles of commonly secreted neuronal factors under KCl, and MDZ conditions. IGF1 is highlighted by a red dashed line. (C) Validation of IGF1and BDNF expression by RT-qPCR. n.s. P >0.05, * P < 0.05, ** P < 0.01, **** P < 0.0001. n=3. (D-E) Representative images and quantification of EdU assay after IGFBP3 experiment in GL261 cells. Scale bar = 100 μm. (F-G) Representative images and quantification of EdU assay after Linsitinib (OSI-906) treated experiments in GL261 cells. Scale bar = 200 μm. (H-I) Activation of the IGF1 downstream PI3K/AKT signaling pathway in glioma cells and its quantitative analysis. (J-K) PI3K/AKT signaling pathway change after IGFBP3 incubation in glioma cells. n.s. P >0.05, * P< 0.05, ** P< 0.01, *** P< 0.001, **** P< 0.0001. n=3.

Article Snippet: The concentrations of IGF1 in Neu-CMs were quantified using a commercial Mouse/Rat IGF1 ELISA Kit (Multi Sciences, China) according to the manufacturer’s instructions.

Techniques: Expressing, Control, Gene Expression, Biomarker Discovery, Quantitative RT-PCR, EdU Assay, Activation Assay, Incubation

Journal: bioRxiv

Article Title: Midazolam suppresses glioma progression by attenuating neuronal activity and downregulating IGF1 signaling

doi: 10.64898/2026.03.31.715727

Figure Lengend Snippet: (A) Overall distribution of IGF1 - related genes in all samples. (B) Standardized expression profile and function annotation of IGF1co-expression genes. (C-D) Over-representation enrichment analysis of genes of interest using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases in KCl treated group and MDZ treated group, respectively. (E) Fuzzy cluster analysis identifying five distinct expression patterns of genes participated in MAPK signaling pathway. (F) Transcriptional activity of Fos was evaluated using a univariable linear model. (G) Predicted c-Fos binding motif within the Igf1 promoter region identified using the JASPAR database. (H) ChIP-qPCR analysis assessing c-Fos enrichment at the predicted regulatory region. * P <0.05, ** P < 0.01, **** P< 0.0001. n=3.

Article Snippet: The concentrations of IGF1 in Neu-CMs were quantified using a commercial Mouse/Rat IGF1 ELISA Kit (Multi Sciences, China) according to the manufacturer’s instructions.

Techniques: Expressing, Activity Assay, Binding Assay, ChIP-qPCR

Journal: Bioactive Materials

Article Title: Sulfated polysaccharide prevents senescent adipocyte-driven osteonecrosis by stem cell fate reprogramming

doi: 10.1016/j.bioactmat.2025.11.039

Figure Lengend Snippet: Schematic illustration of the senescence-regulatory mechanisms of the sulfated polysaccharide in the glucocorticoid-induced bone marrow microenvironment. Bone marrow senescence plays a critical role in the pathogenesis of osteonecrosis. Glucocorticoids act on bone marrow target cells—adipocytes—to initiate primary bone marrow senescence via triggering a positive feedback loop through the prostaglandin/PPARγ/INK signaling axis. Subsequently, these senescent adipocytes propagate SASP factors to adjacent healthy cells through paracrine signaling or direct cell–cell contact, leading to secondary senescence. Sulfated chitosan (SCS) reprograms the lineage commitment bias of LepR + MSCs by activating the IGF-1/PI3K/Akt/mTOR signaling cascade, suppressing adipogenic differentiation and lipid biosynthesis pathways. SCS attenuates the spread of primary adipocyte senescence into secondary senescence, limiting the progressive amplification of the senescence cascade. Ultimately, this strategy halts the onset of senescence-driven osteonecrosis at an early stage and preserves the functional stability of the bone marrow microenvironment.

Article Snippet: Furthermore, to explore the molecular mechanisms by which SCS regulates MSCs lineage bias, bone marrow supernatant was collected on day 7 following co-treatment with SCS and MPS, and ELISA assays for IGF-1 (R&D Systems, MG100) and BMP-2 (R&D Systems, DBP200) were performed as described above.

Techniques: Amplification, Functional Assay

Journal: Bioactive Materials

Article Title: Sulfated polysaccharide prevents senescent adipocyte-driven osteonecrosis by stem cell fate reprogramming

doi: 10.1016/j.bioactmat.2025.11.039

Figure Lengend Snippet: SCS modulates mesenchymal stem cell lineage bias via activation of the IGF-1/PI3K/Akt/mTOR signaling pathway. ( A ) Quantitative analysis of osteocyte morphology in the trabecular bone matrix of the bone marrow at week 6 after MPS treatment with or without SCS, in the presence of various neutralizing antibodies (NAbs) and antagonistic proteins. ( B ) ELISA analysis of IGF-1 and BMP-2 levels in the femoral bone marrow and peripheral serum at day 7 following SCS treatment under MPS conditions. ( C and D ) Western blot analysis of phospho-PI3K, phospho-Akt, and phospho-mTOR (C), as well as phospho-Smad1/5/8, phospho-ERK, and phospho-p38 (D), in CD45 − Ter119 − CD31 − LepR + MSCs after 15-min stimulation with conditioned medium (CM) derived from bone marrow fluid at day 7 following SCS treatment. ( E – G ) Representative flow cytometry plots (E, F) and quantitative analysis (G) of CD45 − CD31 − Sca-1 + CD24 − adipocyte progenitor cells (APCs), CD45 − CD31 − Sca-1 + CD24 + MSCs (E), and CD45 − CD31 − Sca-1 − PDGFRα + (Pα + ) osteoprogenitor cells (OPCs) (F) from femoral bone marrow at day 14 post-MPS induction with or without combined treatment using SCS and IGF-1 NAb or Noggin. ( H and I ) Representative SA-β-Gal staining images (green) of the femur (H), and corresponding quantification (I), at week 4 following MPS treatment with SCS in combination with IGF-1 NAb or DMH1. Insets show magnified views of bone marrow (BM) and trabecular bone matrix (TBM) regions. (Scale bars, 100 μm and 25 μm) ( J ) qPCR analysis of 12 senescence-associated markers in ex vivo femoral bone tissues at week 4 following MPS treatment with SCS in combination with IGF-1 NAb or DMH1. ( K ) Representative Oil Red O staining images of CD45 − Ter119 − CD31 − LepR + MSCs sorted from femurs at day 7 following MPS treatment with SCS in combination with LY294002 or LDN-193189, after in vitro adipogenic induction. (Scale bars, 50 μm and 25 μm) ( L and M ) γ-H2A.X and telomere-associated DNA damage foci (TAFs) co-localization analysis (L), and corresponding quantification (M), in CD45 − Ter119 − CD31 + arteriolar ECs sorted from femurs at day 28 following MPS treatment with SCS in combination with rapamycin or LDN-193189, using immuno-FISH staining. (Scale bars, 7 μm and 1 μm) ( N and O ) Sequential fluorescent labeling using calcein (N) and quantification of mineral apposition rate (O) in femurs treated with SCS and MPS for 4 weeks, with or without LY294002 and/or GW9662. (Scale bars, 50 μm) ( P ) ELISA analysis of five senescence-associated cytokines in femoral bone marrow at day 28 following MPS treatment with SCS in combination with rapamycin and/or T0070907. ( Q and R ) Representative t-distributed stochastic neighbor embedding (t-SNE) plots (Q) from flow cytometric analysis of CD45 − CD31 − Sca-1 + CD24 − APCs, CD45 − CD31 − Sca-1 + CD24 + MSCs, CD45 − CD31 − Sca-1 − Pα + OPCs, CD45 − Ter119 − CD31 + arteriolar ECs, and CD45 − Ter119 − Emcn + sinusoidal ECs at day 14 following MPS treatment with SCS in combination with IGF-1 and/or rosiglitazone, and quantitative analysis of APCs (R) ( S ) Heatmap showing the fluorescent intensity distribution of Lamin-B1 expression across five cellular subpopulations as identified in the t-SNE clustering plot. ∗ P < 0.05 vs. IgG (empty lacunae); # P < 0.05 vs. IgG (filled lacunae). ∗ P < 0.05 vs. SCS; # P < 0.05 vs. SCS + IGF-1 NAb. Data are presented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; ns, not significant. Statistical significance was determined using an unpaired two-tailed Student's t -test ( B ), or one-way ANOVA with Tukey's post hoc test ( A, G, I, J, O, P and R ).

Article Snippet: Furthermore, to explore the molecular mechanisms by which SCS regulates MSCs lineage bias, bone marrow supernatant was collected on day 7 following co-treatment with SCS and MPS, and ELISA assays for IGF-1 (R&D Systems, MG100) and BMP-2 (R&D Systems, DBP200) were performed as described above.

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Derivative Assay, Flow Cytometry, Staining, Ex Vivo, In Vitro, Labeling, Expressing, Two Tailed Test

Journal: Communications Biology

Article Title: Perinatal liver sympathetic innervation governs body size

doi: 10.1038/s42003-026-09880-9

Figure Lengend Snippet: A Representative Tuj1 and DAPI immunohistochemistry (left panel) and quantification of Tuj1 staining (right panel) of liver sections from P7 Nes-WT and Nes-Cdh1 KO mice ( n = 4 per genotype). Scale bar, 160 µm. B Representative TH and DAPI immunohistochemistry (left panel) and quantification of TH staining (right panel) of liver sections from P7 Nes-WT and Nes-Cdh1 KO ( n = 3 per genotype). Scale bar, 160 µm. C Weights of tibialis anterior muscle (TAM), inguinal white adipose tissue (iWAT), and brown adipose tissue (BAT) normalized to body weight in P18 Nes-WT and Nes-Cdh1 KO ( n = 3–4 per genotype). D Representative hematoxylin and eosin staining of TAM sections from P18 Nes-WT and Nes-Cdh1 KO. Scale bar, 200 µm. E Forelimb grip strength measurements show decreased neuromuscular function in Nes-Cdh1 KO mice (P18 n = 3–4 per genotype). F Immunoblot of TH protein from heart, lung, liver and kidney of P7 Nes-WT and Nes-Cdh1 KO. Quantifications are shown in Supplementary Fig. . G Plasma IGF-1 levels of P7 and P21 Nes-WT and Nes-Cdh1 KO ( n = 6 per postnatal period and genotype). H Quantitative real-time PCR of indicated mRNAs in P7 liver from Nes-WT and Nes-Cdh1 KO mice. Data were normalized to Gapdh mRNA levels obtained in the same sample, and relative mRNA levels were considered as the fold change of Nes-WT ( Igf1 n = 9, Igfbp3 n = 7, Igfals n = 9 per genotype). Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01. Shapiro–Wilk normality test and Levene’s equal variances test followed by Welch’s t -test ( A ); unpaired, U-Mann-Whitney’s test ( B , E ); unpaired, two-tailed Student’s t -test ( C ); Welch’s t -test and unpaired U-Mann-Whitney’s test ( G ) or by Welch’s t -test and unpaired, two-tailed Student’s t -test ( H ) versus age-matched Nes-WT.

Article Snippet: Rat/Mouse Growth Hormone ELISA Kit (Millipore #EZRMGH-45K) was used to measure GH plasma levels and Mouse/Rat IGF-I Quantikine ELISA was used to analyze IGF-1 levels (R&D Systems #MG100).

Techniques: Immunohistochemistry, Staining, Western Blot, Clinical Proteomics, Real-time Polymerase Chain Reaction, Two Tailed Test

Journal: Communications Biology

Article Title: Perinatal liver sympathetic innervation governs body size

doi: 10.1038/s42003-026-09880-9

Figure Lengend Snippet: A Hypothalamic-pituitary gland-liver (GHRH-GH-IGF-1) endocrine pathway scheme. Created with Biorender.com. B Representative NeuN (neuronal marker) and DAPI (nuclei) immunohistochemistry of hypothalamic sections (particularly in the arcuate nucleus), at early (P7) and late (P21) postnatal period. Scale bar, 20 µm. Quantifications are shown in Supplementary Fig. . C , D Plasma levels of ( C ) GHRH and ( D ) GH ( n = 4 per postnatal period and genotype). E Representative GH and DAPI immunohistochemistry of pituitary gland sections (P21). Quantifications are shown in Supplementary Fig. . Scale bar, 25 µm. F Quantitative real-time PCR of Gh mRNA in P7 pituitary gland. Data were normalized to Gapdh mRNA levels obtained in the same sample, and relative mRNA levels were considered as the fold change of Nes-WT ( n = 5 per genotype). G – K P7 Nes-Cdh1 KO mice were injected i.p. once daily for 7 days with saline (Nes-WT, Nes-Cdh1 KO) or recombinant human IGF-1 (1 mg/Kg) (Nes-Cdh1 KO + IGF-1). G Body gain (one week, P14-P7), H brain/body weight ratio and I liver/body weight ratio of P14 Nes-WT, Nes-Cdh1 KO and Nes-Cdh1 KO treated with human recombinant IGF-1 (Nes-Cdh1 KO + IGF-1) ( n = 7 per genotype and condition). J Representative Tuj1 and DAPI immunohistochemistry (left panel) and quantification of Tuj1 staining (right panel) of liver sections from P14 Nes-WT, Nes-Cdh1 KO, and Nes-Cdh1 KO + IGF-1 mice ( n = 3 per genotype and condition). Scale bar, 160 µm. K Quantitative real-time PCR of indicated mRNAs in P14 liver. Data were normalized to Gapdh mRNA levels obtained in the same sample, and relative mRNA levels were considered as the fold change of Nes-WT ( Igf1, Igfbp3, Igfals n = 5 per genotype and condition). Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01. Shapiro-Wilk normality test and Levene’s equal variances test followed by Welch’s t -test ( C ); unpaired U-Mann-Whitney’s test and unpaired, two-tailed Student’s t -test ( D ) or by unpaired, two-tailed Student’s t -test ( F ) versus age-matched Nes-WT or two-way ANOVA followed by Bonferroni correction ( G , J , K ) or Welch’s ANOVA test followed by Games-Howell correction ( H , I ).

Article Snippet: Rat/Mouse Growth Hormone ELISA Kit (Millipore #EZRMGH-45K) was used to measure GH plasma levels and Mouse/Rat IGF-I Quantikine ELISA was used to analyze IGF-1 levels (R&D Systems #MG100).

Techniques: Marker, Immunohistochemistry, Clinical Proteomics, Real-time Polymerase Chain Reaction, Injection, Saline, Recombinant, Staining, Two Tailed Test