Journal: Frontiers in pharmacology

Article Title: Volatile Oil from Amomi Fructus Attenuates 5-Fluorouracil-Induced Intestinal Mucositis.

doi: 10.3389/fphar.2017.00786

Figure Lengend Snippet: FIGURE 5 | Reduced IL-6, ROS, TNF-α, NF-κB, and MPO after VOA and BA treatment in blood and small intestinal tissue (n = 6). Increased (A) IL-6 and (B) ROS in blood after 5-FU. VOA and BA reduced inflammation. Increased (C) ROS, (D) TNF-α, (E) MPO, and (F) NF-κB in small intestinal tissue after 5-FU. VOA and BA reduced pro-inflammatory cytokines. Values are means, with their standard deviation represented by vertical bars. Values are means ± SD. ∗Significant difference compared to the controls. # Significant difference compared to the model and controls. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, #p < 0.05, ##p < 0.01, and ###p < 0.001.

Article Snippet: ROS, tumor necrosis factor-α (TNF-α), nuclear factor of kappa b (NF-κB), and myeloperoxidase MPO in the intestinal homogenate were measured with ELISA kits purchased from Cusabio Biotech Co., Ltd. (China).

Techniques: Standard Deviation

Journal: Nature communications

Article Title: VSIG4 inhibits proinflammatory macrophage activation by reprogramming mitochondrial pyruvate metabolism.

doi: 10.1038/s41467-017-01327-4

Figure Lengend Snippet: Fig. 1 Vsig4−/−mice are more susceptible to HFD-induced obesity with insulin resistance. Eight-week-old male Vsig4−/−mice and age-matched C57BL/6 WT controls were fed a HFD. a Body weight was measured and compared. The obese mice were sacrificed after 10 weeks of HFD feeding. b Fat distribution was detected by μCT. Yellow indicates subcutaneous fat and brown indicates that visceral fat. c Measurement of abdominal wall fat, perirenal fat, serum triglyceride, cholesterol, and free fatty acid. d Representative liver H&E staining (left), and intrahepatic triglyceride contents (right), scale bar = 20 μm, n = 10 per group. e Representative the architecture of adipose tissues stained by H&E (left), adipocyte size and cell numbers was calculated (right), scale bar = 20 μm, n = 10 per group. f The 15-h-fasting blood glucose levels and 5-h-fasting serum insulin levels. g GTT and ITT were performed in theses obese mice, n = 6 per group. h Western blot of the AKT, p-Aktser473, and p-IRS-1 in VAT, muscle and liver tissues of obese mice after 4 min of insulin administration, n = 4 per group. ATMs were isolated from obese mice. i Flow cytometry analyzing pro-IL-1β, IFN-γ, and TNF. Cytokines in VAT were detected by j qRT-PCR and k western blot. Error bar, s.e.m. *p < 0.05, **p < 0.01 and ***p < 0.001 (Student’s t-test). Data are representative of five (a) and three (c–i) independent experiments

Article Snippet: ELISA Kits, including TNF (#EK0527), IL-6 (#EK0411), IL-1β (#EK0394), and IL-12 p40 (#EK0932) were from Boster Ltd. (Wuhan, China).

Techniques: Staining, Western Blot, Isolation, Flow Cytometry, Quantitative RT-PCR

Journal: Nature communications

Article Title: VSIG4 inhibits proinflammatory macrophage activation by reprogramming mitochondrial pyruvate metabolism.

doi: 10.1038/s41467-017-01327-4

Figure Lengend Snippet: Fig. 2 Vsig4 deficiency exacerbates MHV-3-induced fulminant hepatitis. The Vsig4−/−mice and age-matched C57BL/6 WT littermates were infected with MHV-3 (100 PFU/mouse) via i.p. injection. a The survival was monitored. b H&E staining of liver, and TUNEL staining of cell apoptosis, scale bar = 20 μm, n = 5–8 per group, arrow indicated positive cells. c Serum ALT and AST levels at 0 h and 48 h post infection (PI), n = 5–8 per group. d Plaque assay of virus titers in livers at 48 h PI. e qRT-PCR analyzing proinflammatory cytokines in PEMs at 12 h and in liver tissues at 72 h of MHV-3 infection. f Flow cytometry analyzing TNF, pro-IL1-β, and IL-6 from PEMs after 12 h of virus infection. g Western blot analyzing proinflammatory cytokines in infected livers at 24 h and 48 h PI, n = 4 per group. h ELISA of serum concentration of proinflammatory mediators, n = 5–10 per group. Error bar, s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001 and NS, p > 0.05. a was analyzed by log-rank test and others are calculated by Student’s t-test. Data are representative of six (a) and three (b–f, h) independent experiments

Article Snippet: ELISA Kits, including TNF (#EK0527), IL-6 (#EK0411), IL-1β (#EK0394), and IL-12 p40 (#EK0932) were from Boster Ltd. (Wuhan, China).

Techniques: Infection, Injection, Staining, TUNEL Assay, Plaque Assay, Virus, Quantitative RT-PCR, Flow Cytometry, Western Blot, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Nature communications

Article Title: VSIG4 inhibits proinflammatory macrophage activation by reprogramming mitochondrial pyruvate metabolism.

doi: 10.1038/s41467-017-01327-4

Figure Lengend Snippet: Fig. 3 VSIG4 impedes LPS-induced macrophage M1 polarization in vitro. PEMs were treated with LPS (2 μg/ml), a qRT-PCR analysis of Il-1β and Tnf transcripts. b ELISA of cytokines in cultured supernatants. c Western blot analyzing cytokine protein expression. d Flow cytometry analyzing surface expression of activation markers. RAW264.7 cells stably infected with lentiviral control vectors (Len-cont.) or vectors encoding Vsig4 (Len-Vsig4), cells were further treated with LPS (2 μg/ml), e qRT-PCR analysis of Il-1β, Il-6, and Tnf transcripts. f ELISA detecting cytokines in cultured supernatant. g Flow cytometry analyzing surface expression of CD40. h C3−/−BMDMs were tranfected to overexpress VSIG4, and cells were further treated with LPS (2 μg/ ml), the secretion of IL-6 and IL-1β was detected by ELISA. Error bar, s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001 and NS, p > 0.05 (Student’s t-test). Data are representative of three independent experiments

Article Snippet: ELISA Kits, including TNF (#EK0527), IL-6 (#EK0411), IL-1β (#EK0394), and IL-12 p40 (#EK0932) were from Boster Ltd. (Wuhan, China).

Techniques: In Vitro, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Culture, Western Blot, Expressing, Flow Cytometry, Activation Assay, Stable Transfection, Infection, Control

Journal: Nature communications

Article Title: VSIG4 inhibits proinflammatory macrophage activation by reprogramming mitochondrial pyruvate metabolism.

doi: 10.1038/s41467-017-01327-4

Figure Lengend Snippet: Fig. 5 VSIG4 triggers PDK2 expression in macrophages. Macrophages from WT and Vsig4−/−mice were collected. a qRT-PCR detection of 4 Pdk isoforms in BMDMs. b Western blot analyzing PDK2, p-PDH-E1αs300, p-PDH-E1αs293, and total PDH. c The location of p-PDH-E1αs300 in mitochondria was analyzed by immunofluoresence double staining, scale bar = 20μm. d Western blot of PDK2, p-PDH-E1αs300, p-PDH-E1αs293 in liver tissues at 0 h and 48 h PI. RAW264.7 cells were transfected to expression of Vsig4, and cells were further treated with LPS (2 μg/ml), e Western blot analysis of PDK2 and p-PDH- E1αs300. f PDH activity analysis, n = 6 per group. The expression of Pdk2 in RAW264.7 cells was silenced by shRNA or enhancing Pdk2 expression by lentivirus infection. g Seahorse analysis of OCR after 2 h of LPS treatment (up), and basal and maximal OCR of the indicated conditions was plotted in bar graphs (down), n = 5 per group. h Flow cytometric assay of mtROS secretion after LPS administration. i ELISA of IL-6 and TNF in cultured supernatants, n = 4 per group. j Flow cytometric assay of LPS-caused CD40 expression at 6 h. Error bar, s.e.m. *p < 0.05,**p < 0.01, ***p < 0.001 and NS, p > 0.05 (Student’s t-test). Data are representative of three independent experiments

Article Snippet: ELISA Kits, including TNF (#EK0527), IL-6 (#EK0411), IL-1β (#EK0394), and IL-12 p40 (#EK0932) were from Boster Ltd. (Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Double Staining, Transfection, Activity Assay, shRNA, Infection, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Nature communications

Article Title: VSIG4 inhibits proinflammatory macrophage activation by reprogramming mitochondrial pyruvate metabolism.

doi: 10.1038/s41467-017-01327-4

Figure Lengend Snippet: Fig. 8 Forced overexpression of Vsig4 improves MHV-3-induced hepatitis. C57BL/6 WT mice were infected with lentivirus (107 PFU/mouse) to induce the expression of Vsig4 in vivo, these mice were further infected with MHV-3 at day 6. a Liver Vsig4 gene transcription was analyzed by qRT-PCR at day 6, n = 5 per group. b Western blotting for PDK2, p-PDH-E1αs300, FGL2, and proinflammatory cytokines TNF, IL-6 and IL-1β in liver tissues at 72 h of MHV-3 infection, n = 4 per group. c The architecture of the liver tissues at 72 h of infection was compared by H&E staining, scale bar = 20 μm, n = 5 per group. d The survival was monitored for a total of 20 days. Error bar, s.e.m. a *p < 0.05 was analyzed by Student’s t-test, and d was analyzed by log-rank test. Data are representative of three independent experiments

Article Snippet: ELISA Kits, including TNF (#EK0527), IL-6 (#EK0411), IL-1β (#EK0394), and IL-12 p40 (#EK0932) were from Boster Ltd. (Wuhan, China).

Techniques: Over Expression, Infection, Expressing, In Vivo, Quantitative RT-PCR, Western Blot, Staining

Journal: Frontiers in Neuroscience

Article Title: Autism-Like Behavior in the Offspring of CYP11A1 -Overexpressing Pregnant Rats

doi: 10.3389/fnins.2021.774439

Figure Lengend Snippet: Primary microglial transcriptome and primary culture indicates immune activation in offspring from CYP11A1 -overexpressing dams. (A) Microglial cells from the offspring of AD-CYP11A1-treated pregnant rats were identified by their Cd11b positivity. (B) GO analysis revealed the activation of immune responses in primary microglia from the CYP11A1 overexpression group. (C) KEGG analysis indicated that GABAergic synapse related genes were enriched from the CYP11A1 overexpression group (over_represented p = 0.033). N = 3 for each group, and only male rats were used for the transcriptomic analysis. (D,E) Expression of the inflammatory tumor necrosis factor-alpha (TNF-α) was increased in the primary microglia in the CYP11A1 adenovirus infected cells. ELISA revealed TNF-α in the adenovirus infected culture medium (D) , and real-time PCR detected TNF-α mRNA in the microglia (E) (* P < 0.05).

Article Snippet: TNF-α levels were measured by rat TNF-α ELISA Kit (D731168, R&D Systems).

Techniques: Activation Assay, Over Expression, Expressing, Infection, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction