Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Hydrangea serrata extract attenuates PM-exacerbated airway inflammation in the CARAS model by modulating the IL-33/ST2/NF-κB signaling pathway.

doi: 10.1016/j.biopha.2024.116596

Figure Lengend Snippet: Fig. 6. HSE regulated IL-4, IL-5, and histamine in PM-exacerbated CARAS mice. (A,B) Levels of IL-4 and IL-5 in NLF. (C,D) Levels of IL-4 and IL-5 in BALF. (E) Levels of histamine in BALF. (G) Levels of IL-33 in BALF. All levels were determined using ELISA kits. Values are presented as mean ± SEM. Significant differences at ###p<0.001 and ##p<0.01 compared with the control group; ***p<0.001, **p<0.01, and *p<0.05 compared with the CARAS/PM group. CARAS, combined allergic rhinitis and asthma syndrome; Dexa, dexamethasone; HSE, Hydrangea serrata extract; OVA, ovalbumin; PM, particulate matter.

Article Snippet: Thus, IL-33 in BALF was measured using an ELISA kit (M33000, R&D Systems Inc., Minneapolis, MN, USA) according to the manufacturer’s guidelines.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: The Journal of Experimental Medicine

Article Title: The transcription factor Bhlhe40 is a switch of inflammatory versus antiinflammatory Th1 cell fate determination

doi: 10.1084/jem.20170155

Figure Lengend Snippet: Bhlhe40 suppresses IL-10 production by Th1 cells. (A) Gene expression values (RPKM) of Il10, Maf , and Ikzf3 from RNA-Seq analysis of C57BL/6 WT and Bhlhe40 cKO Th1 cells ( n = 2). (B) Sorted naive CD4 T cells cultured under Th1-polarizing conditions with plate-bound anti-CD3/CD28 for 4 d and restimulated with PMA-ionomycin for 4 h. Flow cytometric analysis of IFN-γ/IL-10 production (top) and IL-10 expression (bottom left) by CD4 + CD44 hi WT and cKO Th1 cells. Gray solid, WT; red line, cKO. Bottom right: IL-10 production from three independent experiments (≥2 mice per group). *, P < 0.05; **, P < 0.01; ***, P < 0.001; Student’s t test. (C) Sorted naive CD4 T cells cultured under indicated conditions with plate-bound anti-CD3/CD28 for 4 d and restimulated with PMA-ionomycin for 4 h. Flow cytometric analysis of IFN-γ/IL-10 production by CD4 + CD44 hi WT and Bhlhe40 cKO cells. Data are representative of two independent experiments (≥2 mice per group).

Article Snippet: Total PEC samples were cultured with T. gondii –soluble tachyzoite antigen (STAg, 5 µg/ml) in complete RPMI for 3 d. The samples were centrifuged and the supernatant was used to measure the levels of IFN-γ and IL-10 using mouse Quantikine ELISA kits for IFN-γ and IL-10 (R&D systems).

Techniques: Expressing, RNA Sequencing Assay, Cell Culture

Journal: The Journal of Experimental Medicine

Article Title: The transcription factor Bhlhe40 is a switch of inflammatory versus antiinflammatory Th1 cell fate determination

doi: 10.1084/jem.20170155

Figure Lengend Snippet: Bhlhe40 expression in T cells is required for colitis induction. (A) Rag1 −/− mice received sorted naive CD4 + CD25 − CD45RB hi T cells from either WT (red dots) or Bhlhe40 cKO (black squares) or injected i.v. with PBS (blue diamonds) as the control group. Mice were weighed weekly. Statistical significance of body weight of Bhlhe40 cKO versus WT (mean ± SEM, n = 5) at different time points was determined by a two-tailed unpaired Student’s t test. Data are representative of three independent experiments. (B) Graphical representation of the absolute number of CD4 T cells harvested from spleen and mesenteric lymph node (MLN) of Rag1 −/− mice, in which sorted naive CD4 + CD25 − CD45RB hi T cells from either WT (dots) or Bhlhe40 cKO mice (squares) were i.v. transferred for 4 wk ( n = 3 per group) are shown. Data are representative of two independent experiments. (C) Percentages of IFN-γ + and IL-17A + cells among the CD4 + CD44 hi T cells in spleen and MLN from Rag1 −/− mice received WT (dots) or Bhlhe40 cKO transfer (squares) cells for 8 wk (mean ± SEM, n = 5, right). Data are representative of three independent experiments. (D) Sorted naive CD4 T cells from WT or Bhlhe40 cKO were transferred into Rag1 −/− mice. 2 wk after transfer, CD4 T cells were sorted from the spleens and RNAs were prepared from sorted cells. Real time PCR analysis of Ifng and Il10 mRNA was performed (mean ± SEM, n = 5). Data are representative of two independent experiments. (E) Sorted naive CD4 T cells from C57BL/6 or Bhlhe40 cKO were transferred into Rag1 −/− mice. 2 wk after transfer, cells from MLN were restimulated with PMA plus ionomycin, and then intracellular staining for IFN-γ and IL-10 was performed. Flow plots were gated on CD4 + CD44 hi cells. ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001; Student’s t test. Data are representative of two independent experiments.

Article Snippet: Total PEC samples were cultured with T. gondii –soluble tachyzoite antigen (STAg, 5 µg/ml) in complete RPMI for 3 d. The samples were centrifuged and the supernatant was used to measure the levels of IFN-γ and IL-10 using mouse Quantikine ELISA kits for IFN-γ and IL-10 (R&D systems).

Techniques: Expressing, Injection, Two Tailed Test, Real-time Polymerase Chain Reaction, Staining

Journal: The Journal of Experimental Medicine

Article Title: The transcription factor Bhlhe40 is a switch of inflammatory versus antiinflammatory Th1 cell fate determination

doi: 10.1084/jem.20170155

Figure Lengend Snippet: Bhlhe40 cKO mice are susceptible to T. gondii infection. (A) Bhlhe40 WT and cKO mice were infected i.p. with a mean of 15–20 ME49 cysts of T. gondii . Survival of infected mice was monitored. The survival curves shown are one representative of two independent experiments preformed ( n = 11 per group). (B) T. gondii –infected mice were treated i.p. with anti–IL-10R or control IgG1 at 2 d before and 2 d after infection. Survival of infected mice (WT with control IgG1, dots; cKO with control IgG1, squares; cKO with anti–IL-10R, diamonds) was monitored daily ( n = 7–8). The data shown are the pooled results from two independent experiments. (C) T. gondii –infected mice were treated i.p. with anti–IL-10R or control IgG1 at 2 d before and 2 d after infection. Parasite burden was determined by counting infected cells in cytospin smears obtained from PECs on day 8 (mean ± SEM, n = 3–6). *, P < 0.05; Student’s t test.Data are representative of two independent experiments, and similar results were obtained in a third experiment analyzed on day 13. (D) PECs from 13-d infected mice were stained with tetramer to assess the frequency of T. gondii AS15 peptide-specific CD4 T cells. These PECs were also cultured with STAg for 3 d. IFN-γ and IL-10 production by T. gondii antigen-specific cells were measured by ELISA (mean ± SEM, n = 3–6). ns, not significant; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; Student’s t test. Data are representative of two independent experiments.

Article Snippet: Total PEC samples were cultured with T. gondii –soluble tachyzoite antigen (STAg, 5 µg/ml) in complete RPMI for 3 d. The samples were centrifuged and the supernatant was used to measure the levels of IFN-γ and IL-10 using mouse Quantikine ELISA kits for IFN-γ and IL-10 (R&D systems).

Techniques: Infection, Staining, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: N-acetylcysteine attenuates lipopolysaccharide-induced impairment in lamination of Ctip2-and Tbr1- expressing cortical neurons in the developing rat fetal brain

doi: 10.1038/srep32373

Figure Lengend Snippet: (a) The IL-6 levels in amniotic fluid was detected by ELISA. The results show that intra-uterine LPS application caused significantly increased of amniotic IL-6 at highest does (25 μg/kg), and addition of NAC reduces this upregulation. n = 3 assays, each performed in duplicate. (b) Cm-H 2 DCFDA detection kit was used to determine the LPS-induced ROS generation in amniotic fluid. The highest concentration of LPS (25 μg/kg) dramatically induced ROS generation in amniotic fluid, and addition of NAC decreased this outcome. n = 3 assays, each performed in duplicate. (c) The number of embryo was decreased in response to LPS exposure. n values (number of pregnant female rats) for 0 μg/kg LPS = 11; 0.25 μg/kg LPS = 3; 2.5 μg/kg LPS = 5; 12.5 μg/kg LPS = 5; 25 μg/kg LPS = 5; 25 μg/kg LPS plus NAC = 5. (d) Representative Western blot of antioxidative markers in extract from five independent experiments is shown. (e) Quantitative analysis of SOD/tubulin band intensity shows an increase expression of SOD in a dose dependent manner, and the application of NAC eases the effect. n = 5. (f) Quantitative analysis of catalase /tubulin band intensity shows that high dose of LPS significantly induces catalase expression, and NAC hinders the act of LPS. n = 5. (g) Quantitative analysis of HO-1/tubulin band intensity shows an up-regulation of HO-1 in response to LPS, and addition of NAC dose-dependent reduces the HO-1 expression. *Statistically different from vehicle condition without NAC. # Statistically different from each LPS dosage compared to NAC. *p < 0.05, **p < 0.01 , ***p < 0.001 by ANOVA with Tukey-Kramer Multiple Comparisons Test. n = 5.

Article Snippet: IL-6 was evaluated using a Rat IL-6 ELISA Kits (R&D Systems, Minneapolis, MN, USA), following the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Western Blot, Expressing

Journal: Cell & Bioscience

Article Title: The mechanisms by which hypothalamic neuroinflammation induced by neonatal cerebral ischemia–hypoxia leads to decreased thymic function via the HPA axis

doi: 10.1186/s13578-026-01543-w

Figure Lengend Snippet: HI induced the activations of TLR4/HMGB1/NF-κB signaling and microglia, leading to neuroinflammation in hypothalamus. A – B TNF-α and IL-1β on days 1, 3, 7, 14 and 21 after HI, n = 5–8. C Double immunofluorescence staining of Iba-1 and CD68. Scale bar = 20 μm. D Number of Iba-1 positive cells ( n = 4). E CD68 + / Iba-1 + cells rate ( n = 4). F – J CD11b, HMGB1, TLR4, p-P65-NF-κB and P65-NF-κB protein levels on day 7 after HI, n = 5. Data were expressed as the mean ± SD. Comparisons between the two groups were made using an unpaired T-test, * P < 0.05, ** P < 0.01, *** P < 0.001 versus Sham group.

Article Snippet: Serum hormone levels, along with hypothalamic tissue hormone and cytokine levels, were measured using CRH, ACTH, and CORT ELISA kits (Omnimabs, USA), as well as TNF-α and IL-1β ELISA kits (Proteintech, China), following the manufacturers’ instructions.

Techniques: Double Immunofluorescence Staining