Journal: ACS Omega

Article Title: β‑Ecdysone-Loaded Gelatin Methacryloyl Hydrogel Accelerates Diabetic Wound Healing by Improving Angiogenesis

doi: 10.1021/acsomega.5c11972

Figure Lengend Snippet: RT-qPCR analysis of angiogenesis-related and inflammatory gene expression to evaluate the regulatory effects of β-E on HUVECs under high-glucose conditions. (A–C) Expression of angiogenesis-related genes VEGF, FLK1 and FGF1. (D–F) Expression of inflammatory genes TNF-α, IL-6, and IL-1. (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and n = 3).

Article Snippet: β-E (20-hydroxyecdysone; purity 99.20%; #HYN6979) was purchased from MedChemExpress (Shanghai, China).

Techniques: Quantitative RT-PCR, Gene Expression, Expressing

Journal: ACS Omega

Article Title: β‑Ecdysone-Loaded Gelatin Methacryloyl Hydrogel Accelerates Diabetic Wound Healing by Improving Angiogenesis

doi: 10.1021/acsomega.5c11972

Figure Lengend Snippet: Physicochemical characterization of the β-E/GelMA hydrogel. (A) Photograph showing the β-E/GelMA hydrogel before and after UV light exposure. (B) Representative SEM images and macroscopic images of GelMA and β-E/GelMA hydrogels. Scale bar: 20 μm. (C) FTIR spectrum of the hydrogels. (D) Swelling behavior of the hydrogels quantified by swelling ratio. (E, F) Stress–strain curves of hydrogels. (G) In vitro degradation profiles of the hydrogels expressed as degradation ratio (DR). (H) Cumulative release profile of β-E from the β-E/GelMA hydrogel under in vitro conditions (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and n = 3).

Article Snippet: β-E (20-hydroxyecdysone; purity 99.20%; #HYN6979) was purchased from MedChemExpress (Shanghai, China).

Techniques: In Vitro

Journal: ACS Omega

Article Title: β‑Ecdysone-Loaded Gelatin Methacryloyl Hydrogel Accelerates Diabetic Wound Healing by Improving Angiogenesis

doi: 10.1021/acsomega.5c11972

Figure Lengend Snippet: Biocompatibility and bioactivity of the β-E/GelMA hydrogel. (A) Cell viability of HUVECs assessed by the CCK-8 assay following treatment with hydrogel extracts. (B) Live/Dead fluorescence staining of HUVECs cultured under 2D conditions (scale bar: 100 μm). (C) Phalloidin/DAPI staining showing cytoskeletal organization and nuclear morphology of HUVECs (scale bar: 50 μm). (D, E) Transwell migration assay results with corresponding quantitative analysis demonstrating enhanced migratory capacity of HUVECs treated with hydrogel extracts. (F, G) Representative images and quantitative analysis of in vitro Matrigel tube formation by HUVECs, including total tube length (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and n = 3).

Article Snippet: β-E (20-hydroxyecdysone; purity 99.20%; #HYN6979) was purchased from MedChemExpress (Shanghai, China).

Techniques: CCK-8 Assay, Fluorescence, Staining, Cell Culture, Transwell Migration Assay, In Vitro

Journal: ACS Omega

Article Title: β‑Ecdysone-Loaded Gelatin Methacryloyl Hydrogel Accelerates Diabetic Wound Healing by Improving Angiogenesis

doi: 10.1021/acsomega.5c11972

Figure Lengend Snippet: β-E/GelMA hydrogels promote diabetic wound healing in vivo. (A) Schematic illustration of hydrogel application to diabetic skin wounds. (B) Digital photographs of the wound areas on day 0, 3, 6, 9, and 12 with the administration of the normal saline (Con and Glu groups), GelMA, and β-E/GelMA. (C) Quantitative outlines showing changes in wound area over time. (D) Statistical analysis of wound closure rates (%) ( n = 3). (E) H&E staining of wound tissues collected on day 12 postinjury (upper scale bar: 500 μm; lower scale bar: 50 μm; black arrows indicate wound margins). (F) Masson’s trichrome staining of wound sections on day 12 postinjury (upper scale bar: 500 μm; lower scale bar: 50 μm; black arrows indicate wound margins). (G–J) Immunohistochemical staining of CD31 and α-SMA in wound tissues on day 12, along with the corresponding quantitative analyses (scale bar: 50 μm, n = 5). (* P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.).

Article Snippet: β-E (20-hydroxyecdysone; purity 99.20%; #HYN6979) was purchased from MedChemExpress (Shanghai, China).

Techniques: In Vivo, Saline, Staining, Immunohistochemical staining

Journal: Science Advances

Article Title: Opposing insulin signals underlie the same developmental switch across hemipteran insects

doi: 10.1126/sciadv.aea4413

Figure Lengend Snippet: ( A ) Identification of ecdysteroids as a target of IIS by comparative transcriptomics. Fifth-instar Wt SW (short-winged–destined) and PaInR2 De4 (long-winged–destined) P. apterus nymphs, and fifth-instar Wt SW (short-winged–destined) and NlInR2 E4 (long-winged–destined) N. lugens nymphs were collected for RNA sequencing (RNA-seq). Three KEGG pathways are commonly regulated in the context of inactivation of IIS in the firebug (via PaInR2 De4 ) and activation of IIS in the planthopper (via NlInR2 E4 ). The ecdysteroid biosynthesis pathway is outlined on the right. ( B ) Heatmap of gene regulation by PaInR2 De4 . Three genes key to ecdysteroid biosynthesis are down-regulated in the context of IIS inactivation (via PaInR2 De4 ). ( C ) Heatmap of gene regulation by NlInR2 E4 . Three genes key to ecdysteroid biosynthesis are down-regulated in the context of IIS activation (via NlInR2 E4 ). ( D ) PaInR2 De4 firebugs decrease titers of the ecdysteroid makisterone A. Fifth-instar nymphs at 1, 3, 5, 7, and 9 days (d) after eclosion were collected for makisterone A determination. The duration of fifth-instar PaInR2 De4 and Wt SW nymphs is indicated at the bottom. ( E ) NlInR2 E4 planthoppers decrease titers of the ecdysteroid 20E. Fifth-instar nymphs at 2, 24, 48, and 72 hours (h) after eclosion were collected for 20E determination. The duration of fifth-instar PaInR2 De4 and Wt SW nymphs is indicated at the bottom. Bars in (D) and (E) represent means ± SEM derived from three independent biological replicates. * P < 0.05, ** P < 0.01, and *** P < 0.001 (two-tailed unpaired t test).

Article Snippet: Standard 20E and makisterone A were purchased from MedChemExpress (HY-N6979) and Macklin (M912023), respectively.

Techniques: Transcriptomics, RNA Sequencing, Activation Assay, Derivative Assay, Two Tailed Test

Journal: Science Advances

Article Title: Opposing insulin signals underlie the same developmental switch across hemipteran insects

doi: 10.1126/sciadv.aea4413

Figure Lengend Snippet: ( A ) Wing morphs of PaInR2 De4 after makisterone A microinjection. ** P < 0.01 and *** P < 0.001 (Pearson’s χ 2 test: χ 2 = 7.501 and df = 1 for 0.1 μg/μl and χ 2 = 63.188 and df = 1 for 0.5 μg/μl). ( B ) Phenotypes of PaInR2 De4 microinjected with DMSO (long-winged morphs). ( C ) Phenotypes of PaInR2 De4 microinjected with makisterone A (short-winged morphs). ( D ) Wing morphs of NlInR2 E4 after 20E microinjection. *** P < 0.001 (Pearson’s χ 2 test: χ 2 = 40.675 and df = 1 for 0.2 μg/μl, χ 2 = 32.125 and df = 1 for 1 μg/μl, and χ 2 = 32.178 and df = 1 for 2.5 μg/μl). ( E ) Phenotypes of NlInR2 E4 microinjected with DMSO (long-winged morphs). ( F ) Phenotypes of NlInR2 E4 microinjected with 20E (short-winged and intermediate size–winged morphs). Forewings and hindwings are indicated by arrows and arrowheads, respectively.

Article Snippet: Standard 20E and makisterone A were purchased from MedChemExpress (HY-N6979) and Macklin (M912023), respectively.

Techniques: Microinjection

Journal: Science Advances

Article Title: Opposing insulin signals underlie the same developmental switch across hemipteran insects

doi: 10.1126/sciadv.aea4413

Figure Lengend Snippet: The IIS pathway acts as both a positive and a negative regulator to control ecdysteroid titers in P. apterus and N. lugens via the InRs-Chico-PDK1-Tsc1/2-TORC1 and InRs-Chico-PDK1-Akt-FoxO cascades, respectively. In P. apterus , inactivation of the IIS pathway decreases ecdysteroid (makisterone A) biosynthesis via targeting Torso , Phantom , and Disembodied genes. In N. lugens , activation of the IIS pathway decreases ecdysteroid (20E) biosynthesis via targeting Neverland , Spook , and Cyp314A1 genes. Low ecdysteroid titers may guide the bipotential wing buds to develop into long wings. In contrast, high levels of ecdysteroid titers may guide wing buds to develop into short wings. Down and up arrows in colors indicate inactivation and activation, respectively.

Article Snippet: Standard 20E and makisterone A were purchased from MedChemExpress (HY-N6979) and Macklin (M912023), respectively.

Techniques: Control, Activation Assay