Journal: PLoS Pathogens

Article Title: H1N1 influenza virus infection results in adverse pregnancy outcomes by disrupting tissue-specific hormonal regulation

doi: 10.1371/journal.ppat.1006757

Figure Lengend Snippet: Hormone expression in sera was quantified via ELISA in uninfected and infected non-pregnant mice for A. progesterone; C. prostaglandin F2α (PGF2α); and D. prostaglandin E2 (PGE2). Cytokine and chemokine expression in sera was quantified via Bio-Rad 23-plex assay in B. uninfected pregnant (E16) (N = 5) and non-pregnant mice (N = 5); E. uninfected and infected pregnant (E16; 4 d.p.i) (N = 5 per group); and F. infected (4 d.p.i) non-pregnant and pregnant (E16) mice (N = 5 per group). Student’s t-test was performed between selected groups and significance noted above asterisk brackets. Inf.: infected; Uninf: uninfected; NP: non-pregnant; P: pregnant mice.

Article Snippet: Progesterone, prostaglandin F2α (PGF2α) and prostaglandin E2 (PGE2) were quantified via ELISA kits from ALPCO (Salem, NH).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Infection, Plex Assay

Journal: PLoS Pathogens

Article Title: H1N1 influenza virus infection results in adverse pregnancy outcomes by disrupting tissue-specific hormonal regulation

doi: 10.1371/journal.ppat.1006757

Figure Lengend Snippet: Hormone expression in lung lysates was quantified via ELISA in uninfected (N = 6) and infected (N = 5) non-pregnant and pregnant mice for A. progesterone; B. prostaglandin F2α (PGF2α); and C. prostaglandin E2 (PGE2). Cytokine and chemokine expression in sera was quantified in lung lysates of D. uninfected pregnant (E16) (N = 8) and non-pregnant mice (N = 6); E. infected pregnant (4 d.p.i., E16) (N = 5) and non-pregnant mice (N = 5). Student’s t-test was performed between selected groups and significance noted above asterisk brackets. Abbreviations for groups are as described in .

Article Snippet: Progesterone, prostaglandin F2α (PGF2α) and prostaglandin E2 (PGE2) were quantified via ELISA kits from ALPCO (Salem, NH).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Infection

Journal: PLoS Pathogens

Article Title: H1N1 influenza virus infection results in adverse pregnancy outcomes by disrupting tissue-specific hormonal regulation

doi: 10.1371/journal.ppat.1006757

Figure Lengend Snippet: Placental lysates were isolated from uninfected and infected pregnant mice (E16). Hormone expression was quantified via ELISA for A. progesterone (N = 6 per group); B. prostaglandin F2α (PGF2α) (N = 9, uninfected, N = 7, infected) and C. prostaglandin E2 (PGE2) (N = 9 per group). Student’s t-test was performed between selected groups and significance noted above asterisk brackets.

Article Snippet: Progesterone, prostaglandin F2α (PGF2α) and prostaglandin E2 (PGE2) were quantified via ELISA kits from ALPCO (Salem, NH).

Techniques: Isolation, Infection, Expressing, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Impact of Mechanical Load on the Expression Profile of Synovial Fibroblasts from Patients with and without Osteoarthritis

doi: 10.3390/ijms20030585

Figure Lengend Snippet: Impact of static compressive force application on the expression of proinflammatory genes. Gene expression and protein secretion of ( a ) TNF-α, ( b ) COX-2/PG-E2 and ( c ) IL-6 of N-SF and OA-SF after 48 h with or without static compressive force application. AU: arbitrary units; RT-qPCR: n = 9; ELISA: n = 6. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤0.001. Statistics: Welch-corrected ANOVA with Games-Howell post-hoc-tests.

Article Snippet: For quantification of tumour necrosis factor α (TNFα) and prostaglandin E2 (PGE2), interleukin-6 (IL-6), collagen-1 (COL-1) and fibronectin (FN-1) protein secretion into the synovial fibroblast supernatant, we used commercially available ELISA kits according to the manufacturers’ instructions (TNFα: EK0525, Boster Biological Technology; PGE2: 514010; Cayman chemical; IL-6: EK0410 Boster Biological Technology; COL-1: ab210966, Abcam; FN-1: EK0349, Boster Biological Technology).

Techniques: Expressing, Gene Expression, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Journal of Traditional Chinese Medicine

Article Title: Yanghe decoction (阳和汤) attenuated pain hypersensitivity induced by michigan cancer foundation-7 injection in rats with bone metastases from breast cancer by inhibiting transient receptor potential ankyrin 1

doi: 10.19852/j.cnki.jtcm.20220707.001

Figure Lengend Snippet: A: qRT-PCR was conducted to detect the mRNA expression of TRPA1; B: Western blot was conducted to detect the protein expression of TRPA1; C: the level of TRPA1 protein; D: Western blot was conducted to detect the protein expression of TNF-α and IL-6, IL-1β and PGE2; E: the level of TNF-α, IL-6, IL-1β and PEG2 protein. Sham: sham operation treat with PBS; YHD : sham operation of rats treated with 18 g•kg-1•d-1 Yanghe decoction; Metastasis: bone cancer metastasis of rats treated with PBS; Metastasis + YHD: rats were injected with MCF-7 before treated with 18 g • kg-1 • d-1 Yanghe decoction; Metastasis+overexpression (oe)-TRPA1: rats were injected with MCF-7 transfected with pEGFP-TRPA1; Metastasis + oe-TRPA1 + YHD: rats were injected with MCF-7 transfected with pEGFP-TRPA1 before treated with 18 g • kg-1 • d-1 Yanghe decoction. Measurement after 14 d of Yanghe decoction intervention. S: Sham; Y: YHD; M: metastasis; T: oe-TRPA1; TRPA1: transient receptor potential ankyrin 1; YHD: Yanghe decoction; TNF-α: tumor necrosis factor-α; IL-6: interleukin 6; IL-1β: interleukin 1 beta; PGE2: prostaglandin E2; oe: overexpression; DRG: dorsal root ganglion. qRT-PCR: quantitative real-time polymerase chain reaction. aP < 0.05, compared with Sham; bP < 0.05, compared with Metastasis; cP < 0.05, compared with Metastasis + oe-TRPA1. P < 0.05 was considered to be statistically significant. One-way analysis of variance among multiple groups, followed by Tukey’s post-hoc tests.

Article Snippet: The first antibodies were: tumor necrosis factor-α (TNF-α) (60291-1-Ig, 1∶2000, proteintech, Chicago, IL, USA), anti-interleukin 6 (IL-6) (66146-1-Ig, 1∶1000, proteintech, Chicago, IL, USA), anti-IL-1β (ab9722, 1∶1000, Abcam, Cambridge, UK), anti- prostaglandin E2 (PGE2) (Orb110374, 1∶500, Biorbyt, Cambridge, UK), anti-TRPA1 (19124-1-AP, 1∶500, proteintech, Chicago, IL, USA), and anti-β-actin (66009-1-Ig, 1∶5000, proteintech, Chicago, IL, USA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Injection, Over Expression, Transfection, Real-time Polymerase Chain Reaction

Journal: Journal of Traditional Chinese Medicine

Article Title: Yanghe decoction (阳和汤) attenuated pain hypersensitivity induced by michigan cancer foundation-7 injection in rats with bone metastases from breast cancer by inhibiting transient receptor potential ankyrin 1

doi: 10.19852/j.cnki.jtcm.20220707.001

Figure Lengend Snippet: A: IHC staining sections of rat tibia tissue were observed under microscope and representative images were taken for the detection of TNF-α, the signature inflammatory cytokine. A1: IHC staining sections of rat tibia tissue of Sham group. A2: IHC staining sections of rat tibia tissue of YHD group. A3: IHC staining sections of rat tibia tissue of Metastasis + YHD group. A4: IHC staining sections of rat tibia tissue of Metastasis + oe-TRPA1 group. A5: IHC staining sections of rat tibia tissue of Sham group. A6: IHC staining sections of rat tibia tissue of Metastasis + oe-TRPA1 + YHD group. The serum levels of PGE2 (B), The level of TNF-α (C), The level of IL-6 (D), The level of IL-1β (E) from groups measured by ELISA. Sham: sham operation treat with PBS; YHD : sham operation of rats treated with 18 g • kg-1 • d-1 Yanghe decoction; Metastasis: bone cancer metastasis of rats treated with PBS; Metastasis + YHD: rats were injected with MCF-7 before treated with 18 g•kg-1•d-1 Yanghe decoction; Metastasis + overexpression (oe)-TRPA1: rats were injected with MCF-7 transfected with pEGFP-TRPA1; Metastasis+oe-TRPA1 + YHD: rats were injected with MCF-7 transfected with pEGFP-TRPA1 before treated with 18 g • kg-1 • d-1 Yanghe decoction. Measurement after 14 d of Yanghe decoction intervention. TRPA1: transient receptor potential ankyrin 1; YHD: Yanghe decoction; TNF-α: tumor necrosis factor-α; IL-6: interleukin 6; IL-1β: interleukin 1 beta; PGE2: prostaglandin E2; oe: overexpression; IHC: immunohistochemistry; ELISA: enzyme-linked immunosorbent assay. aP < 0.05 compared with Sham; bP < 0.05 compared with Metastasis; cP < 0.05 compared with Metastasis + oe-TRPA1. P < 0.05 was considered to be statistically significant. One-way analysis of variance among multiple groups, followed by Tukey’s post-hoc tests.

Article Snippet: The first antibodies were: tumor necrosis factor-α (TNF-α) (60291-1-Ig, 1∶2000, proteintech, Chicago, IL, USA), anti-interleukin 6 (IL-6) (66146-1-Ig, 1∶1000, proteintech, Chicago, IL, USA), anti-IL-1β (ab9722, 1∶1000, Abcam, Cambridge, UK), anti- prostaglandin E2 (PGE2) (Orb110374, 1∶500, Biorbyt, Cambridge, UK), anti-TRPA1 (19124-1-AP, 1∶500, proteintech, Chicago, IL, USA), and anti-β-actin (66009-1-Ig, 1∶5000, proteintech, Chicago, IL, USA).

Techniques: Immunohistochemistry, Microscopy, Enzyme-linked Immunosorbent Assay, Injection, Over Expression, Transfection

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: Response of chondrocytes to shear stress: antagonistic effects of the binding partners Toll-like receptor 4 and caveolin-1.

doi: 10.1096/fj.11-184861

Figure Lengend Snippet: Figure 3. TLR4 and caveolin-1 exert antagonistic effects on shear-induced IL-6 synthesis in human chondrocytes. T/C-28a2 chondrocytes were subjected to shear stress (20 dyn/cm2) or static conditions (0 dyn/cm2) for 2 h (A, B) or 48 h (C, D). In select experiments, T/C-28a2 chondrocytes were transfected with either a siRNA oligonucleotide sequence specific for TLR4 (A) or caveolin-1 (D) or scrambled siRNA control before their exposure to fluid shear. In separate experiments, cells were transfected with either a plasmid containing the cDNA of caveolin-1 (B) or TLR4 (C) or an empty vector (control) prior to shear exposure. TLR4 (A, C) or caveolin-1 (B, D) and IL-6 protein (top panels) and mRNA expression (bottom panels) were determined by Western blot analysis and qRT-PCR, respectively. -Actin and GAPDH served as internal controls in immunoblotting and qRT-PCR assays, respectively. Western blots are representative of 3 independent experiments, all revealing similar results. Data represent means se of 3 independent qRT-PCR experiments. *P 0.05 vs. static control; ŒP 0.05 vs. shear alone.

Article Snippet: The caveolin-1, TLR4, mPGES-1, and L-PGDS, EP2, EP3 cDNA plasmids were obtained from Origene Technologies (Rockville, MD, USA), and subcloned to the pCMV6-XL vector.

Techniques: Shear, Transfection, Sequencing, Control, Plasmid Preparation, Expressing, Western Blot, Quantitative RT-PCR

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: Response of chondrocytes to shear stress: antagonistic effects of the binding partners Toll-like receptor 4 and caveolin-1.

doi: 10.1096/fj.11-184861

Figure Lengend Snippet: Figure 8. Fluid shear induces the binding of the NF-B p65 subunit to the IL-6 promoter in human chondrocytes. T/C-28a2 chondrocytes were subjected to fluid shear stress (20 dyn/cm2) or static conditions (0 dyn/cm2) for 48 h (A, B, F, G) or 2 h (C–E, H–J). In select experiments, T/C-28a2 cells were transfected with the indicated siRNAs or cDNA constructs before exposure to shear stress. In separate experiments, T/C-28a2 cells were subjected to fluid shear stress (20 dyn/cm2) for 2 h in the presence or absence of a MEK1/2 inhibitor [U0126 (10 M) or PD98095 (20 M)]. A–E) Nuclear extracts were then isolated, and NF-B-specific DNA-protein complex formation was determined by gel shift. F–J) Supershift assays using an anti-p65 Ab were carried out as outlined in Materials and Methods. Results of a competition experiment using 200-fold unlabeled NF-B oligonucleotide (cold probe) are shown. Gels are representative of 3 independent experiments, all revealing similar results.

Article Snippet: The caveolin-1, TLR4, mPGES-1, and L-PGDS, EP2, EP3 cDNA plasmids were obtained from Origene Technologies (Rockville, MD, USA), and subcloned to the pCMV6-XL vector.

Techniques: Shear, Binding Assay, Transfection, Construct, Isolation, Gel Shift

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: Response of chondrocytes to shear stress: antagonistic effects of the binding partners Toll-like receptor 4 and caveolin-1.

doi: 10.1096/fj.11-184861

Figure Lengend Snippet: Figure 9. Fluid shear induces the binding of the NF-B p65 subunit to the IL-6 promoter in human chondrocytes. T/C-28a2 cells were subjected to fluid shear stress (20 dyn/cm2) or static conditions (0 dyn/cm2) for 48 h (A, B) or 2 h (C–G). Cells were transfected with the indicated siRNAs or cDNA constructs or treated with prescribed pharmacological inhibitors before exposure to shear stress. Cross-linked chromatin was immunoprecipitated using an antip65 antibody (left panel). In ChIP assays, the anti-RNA polymerase II antibody was used as a positive control, whereas the normal mouse IgG and anti-TLR4 antibodies were used as negative controls. DNA purified from both the immunoprecipitated (IP) and preimmune (input) specimens were subjected to PCR amplification using primers for the GAPDH (control) and p65 promoter genes. All experiments are representative of 3 independent experiments, all revealing similar results.

Article Snippet: The caveolin-1, TLR4, mPGES-1, and L-PGDS, EP2, EP3 cDNA plasmids were obtained from Origene Technologies (Rockville, MD, USA), and subcloned to the pCMV6-XL vector.

Techniques: Shear, Binding Assay, Transfection, Construct, Immunoprecipitation, Positive Control, Control