Journal: Journal of Reproduction & Infertility

Article Title: Investigating the Effect of Crocus sativus (Saffron) Petal Hydro-alcoholic Extract on Ovarian Follicle, Inflammatory Markers, and Antioxidant Enzymes in Mice Model of Polycystic Ovary Syndrome

doi: 10.18502/jri.v23i1.8447

Figure Lengend Snippet: The plasma level(s) of TNF-α, IL6, IL1ß, IL18, and CRP

Article Snippet: Levels of IL-6, TNF-α, IL-1ß, IL-18, and CRP in all serum samples were determined by special ELISA kits (Accu-Bind, Monobind Inc., USA) ( ).

Techniques:

Journal: Journal of Reproduction & Infertility

Article Title: Investigating the Effect of Crocus sativus (Saffron) Petal Hydro-alcoholic Extract on Ovarian Follicle, Inflammatory Markers, and Antioxidant Enzymes in Mice Model of Polycystic Ovary Syndrome

doi: 10.18502/jri.v23i1.8447

Figure Lengend Snippet: The plasma level(s) of TNF-α, IL6, IL1ß, IL18, and CRP

Article Snippet: Levels of IL-6, TNF-α, IL-1ß, IL-18, and CRP in all serum samples were determined by special ELISA kits (Accu-Bind, Monobind Inc., USA) ( ).

Techniques:

Journal: BioMed Research International

Article Title: A De- O -acylated Lipooligosaccharide-Based Adjuvant System Promotes Antibody and Th1-Type Immune Responses to H1N1 Pandemic Influenza Vaccine in Mice

doi: 10.1155/2016/3713656

Figure Lengend Snippet: Th1-type-predominant immune response to CIA06-adjuvanted influenza vaccine. Splenocytes were isolated from the mice that had been immunized as described in and cultured for 72 h in the absence (■) or presence of Greenflu-S (■). The levels of IFN- γ (a) and IL-5 (b) in the culture media were determined by sandwich ELISA, and IFN- γ : IL-5 ratios were calculated for each group (c). Statistical differences were analyzed by one-way ANOVA followed by Tukey's multiple comparison test. Results are expressed as the means ± SD of values obtained from triplicate cultures that used two spleens each. Data shown are representative of three independent experiments. ∗ P < 0.05; ∗∗ P < 0.01; and ∗∗∗ P < 0.001. NS, not significant.

Article Snippet: Recombinant mouse IL-2 was acquired from R&D Systems (Minneapolis, MN, USA), and IFN- γ and IL-5 cytokine ELISA kits were obtained from BD Biosciences (San Diego, CA, USA) and R&D Systems.

Techniques: Isolation, Cell Culture, Sandwich ELISA, Comparison

Journal: BioMed Research International

Article Title: A De- O -acylated Lipooligosaccharide-Based Adjuvant System Promotes Antibody and Th1-Type Immune Responses to H1N1 Pandemic Influenza Vaccine in Mice

doi: 10.1155/2016/3713656

Figure Lengend Snippet: CIA06-adjuvanted influenza vaccine stimulates both CD4 + and CD8 + T cell responses. Splenocytes were isolated from the mice ( n = 6) that had been immunized twice at a 3-week interval with nonadjuvanted or adjuvanted Greenflu-S (0.2 μ g) and stimulated with the vaccine for 72 h in the absence or presence of anti-CD4 and/or anti-CD8 antibodies. IFN- γ levels in the culture media were determined by sandwich ELISA. Statistical differences were analyzed by one-way ANOVA followed by Tukey's multiple comparison test. Results are expressed as the means ± SD of values obtained from triplicate cultures that used two spleens each. Data shown are representative of three independent experiments. ∗ P < 0.05; ∗∗ P < 0.01; and ∗∗∗ P < 0.001. NS, not significant.

Article Snippet: Recombinant mouse IL-2 was acquired from R&D Systems (Minneapolis, MN, USA), and IFN- γ and IL-5 cytokine ELISA kits were obtained from BD Biosciences (San Diego, CA, USA) and R&D Systems.

Techniques: Isolation, Sandwich ELISA, Comparison

Journal: Journal of Translational Medicine

Article Title: Low-intensity pulsed ultrasound triggers a beneficial neuromodulation in dementia mice with chronic cerebral hypoperfusion via activation of hippocampal Fndc5/irisin signaling

doi: 10.1186/s12967-022-03824-7

Figure Lengend Snippet: Boosting hippocampal Fndc5/irisin alleviate hippocampal damage and cognitive deficits following experimental VaD. A Experimental outline. Forced expression of Fndc5 in hippocampus of adult C57BL/6 mice using adenovirus brain infection, while enhancement of hippocampal irisin level by bilateral intrahippocampal injection of recombinant irisin on day 22 after BCAS. B Hippocampal Fndc5 mRNA expression and irisin were respectively detected using qPCR and ELISA on day 28 after BCAS. C Hippocampal synaptic plasticity was analyzed using Patch Clamp on day 28 after BCAS. The average traces of fEPSP in hippocampal slices. D The data of fEPSP at 120 min. E – G Cognitive evaluation were performed on day 28 after BCAS. The errors (numbers of entries into incorrect maze arms) in two-days radial arm water maze test were continuously measured in each block ( E ). The errors in last block were counted in a histogram ( F ). Exploratory time on old or new object in novel object recognition test were illustrated in histogram ( G ). H , I Hippocampal level of TGF-β and IL-10, were determined using ELISA on day 28 after BCAS. J The percentage of freezing time obtained on day 28 after BCAS in contextual fear test were illustrated in histogram. Sham, sham-operated mice; BCAS, mice were subjected to BCAS injury; AdFndc5 or AdGFP, adult BCAS mice were infected using intracerebroventricular injection with an adenoviral vector designed to express Fndc5 or GFP; Irisin, adult BCAS mice with bilateral intrahippocampal injection of recombinant irisin. (n = 8 per group. All data are expressed as mean ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001, ns means no statistical significance)

Article Snippet: The concentration of irisin in serum, brain tissue homogenate or culture supernatant were detected using the irisin ELISA kit obtained from Phoenix pharmaceuticals (Burlingame, USA). and were obtained from Phoenix pharmaceuticals (Burlingame, USA). transforming growth factor (TGF)-β, and interleukin (IL)-10 were measured with mouse TGF-β and IL-10 ELISA Kits (Aviscera Bioscience, Santa Clara, CA, USA).

Techniques: Expressing, Infection, Injection, Recombinant, Enzyme-linked Immunosorbent Assay, Patch Clamp, Blocking Assay, Plasmid Preparation

Journal: Journal of Translational Medicine

Article Title: Low-intensity pulsed ultrasound triggers a beneficial neuromodulation in dementia mice with chronic cerebral hypoperfusion via activation of hippocampal Fndc5/irisin signaling

doi: 10.1186/s12967-022-03824-7

Figure Lengend Snippet: Fndc5/irisin mediates the neurorestorative effects of LIPUS on experimental VaD. A Experimental outline. Knockdown the expression of hippocampal Fndc5 using shRNA before 7days of BCAS. Transcranial LIPUS was applied 24 h after BCAS and subsequently daily with a stimulation time of 5 min at an ultrasound pressure of 0.51 MPa for a period of 28 days. B , C Fndc5 mRNA expression and irisin concentration in hippocampus of adult mice were respectively detected using qPCR and ELISA on day 28 after BCAS. D Hippocampal synaptic plasticity was analyzed using Patch Clamp, average traces of fEPSP in hippocampal slices collected on day 28 after BCAS. E The data of fEPSP at 120 min. F , G Hippocampal TGF-β and IL-10 were determined using ELISA on day 28 after BCAS. H Exploratory time on old or new object in novel object recognition test were illustrated in histogram. I The errors in radial arm water maze test were continuously measured in each block on day 28 after BCAS. J The data of errors in last block were counted in a histogram. K Global Fndc5 knock-out mice (F5KO) was employed to mechanism exploration, contextual fear test was used to assess cognitive deficit on day 28 after BCAS. the percentage of freezing time obtained on day 28 after BCAS were illustrated in histogram. Sham, sham-operated mice; BCAS, mice were subjected to BCAS injury; LIPUS, BCAS adult mice with LIPUS treatment; shFndc5 or shLuc, LIPUS-treated adult BCAS mice with intrahippocampal injection with lentiviral particles expressing shRNA against murine Fndc5 or luciferase. (n = 8 per group. All data are expressed as mean ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001, ns means no statistical significance)

Article Snippet: The concentration of irisin in serum, brain tissue homogenate or culture supernatant were detected using the irisin ELISA kit obtained from Phoenix pharmaceuticals (Burlingame, USA). and were obtained from Phoenix pharmaceuticals (Burlingame, USA). transforming growth factor (TGF)-β, and interleukin (IL)-10 were measured with mouse TGF-β and IL-10 ELISA Kits (Aviscera Bioscience, Santa Clara, CA, USA).

Techniques: Expressing, shRNA, Concentration Assay, Enzyme-linked Immunosorbent Assay, Patch Clamp, Blocking Assay, Knock-Out, Injection, Luciferase

Journal: Advanced Science

Article Title: Extracellular Vesicles Derived from Ligament Tissue Transport Interleukin‐17A to Mediate Ligament‐To‐Bone Crosstalk in Ankylosing Spondylitis

doi: 10.1002/advs.202406876

Figure Lengend Snippet: Pathological ossification and inflammatory activation in non‐ossified ligaments from patients with AS. A). µCT images show the 3D reconstruction and spinal ligament tissue from patients with AS. The red pentagram indicates uncalcified ligament. B). SOFG and HE staining of the AS patient ligament sections showing osteogenic differentiation and disorganized arrangement of collagen fibers. C,D). Representative immunohistochemistry and immunofluorescence staining images showing over‐expression of RUNX2, CD68, and PDGFRα in AS spinal ligament tissue. E). Heatmap of differentially upregulated genes, regulation of osteoblast differentiation (GO:0045667), and cytokine‐mediated signaling pathway (GO:0019221). F). Quantitative analysis of the immunofluorescence area occupied by stained regions in the different groups in (D). G). Concentration of CXCL12 in AS ligament tissue measured using ELISA. H). Pearson's correlation analysis of PDGFRα expression and the expression of markers of AS‐related osteoblast differentiation (RUNX2 and CXCL12) and inflammation (CD68) based on the ELISA and immunofluorescence staining results. I). Left: experimental scheme of AS or HD ligament tissue co‐culture with AS‐MSCs. Middle: ARS staining of AS‐MSCs after 21 days of osteogenic induction. Right: quantitative analysis of the ARS staining (absorption, 562 nm). n = 5 per group. J). Experimental scheme of AS or HD ligament tissue co‐culture with AS‐BMMCs. Concentration of inflammation factors after co‐culturing AS or HD ligament tissue with AS‐BMMCs measured using ELISA. IL, interspinous ligament; SL, supraspinous ligament; SP, spinous process; HE, hematoxylin‐eosin staining; SOFG, Safranin O–Fast Green staining.

Article Snippet: The following ELISA kits were used according to the manufacturer's protocol: CXCL12 (Solarbio, China), TNF‐α (Solarbio, China), IL‐1β (Solarbio, China), IL‐6 (Solarbio, China), and MMP14 (Thermo Scientific, USA).

Techniques: Activation Assay, Staining, Immunohistochemistry, Immunofluorescence, Over Expression, Concentration Assay, Enzyme-linked Immunosorbent Assay, Expressing, Co-Culture Assay

Journal: Advanced Science

Article Title: Extracellular Vesicles Derived from Ligament Tissue Transport Interleukin‐17A to Mediate Ligament‐To‐Bone Crosstalk in Ankylosing Spondylitis

doi: 10.1002/advs.202406876

Figure Lengend Snippet: AS‐LTEVs facilitate inflammation activation by delivering IL‐17A in vivo. A). Diagrammatic representation of experimental strategies for macrophage polarization. B). Representative flow cytometry results of M1 macrophages after treatment with PBS, HD‐LTEVs, AS‐LTEVs, AS‐LTEVs with secukinumab, AS‐LTEVs with secukinumab, and Dynasore. Percentages indicate the proportion of M1 macrophages. n = 3 per group. C). Representative flow cytometry results of DCFH‐DA after treatment with PBS, HD‐LTEVs, AS‐LTEVs, AS‐LTEVs with secukinumab, AS‐LTEVs with secukinumab, and Dynasore. The right panel shows the quantitative analysis of the positive cell rate of ROS in the different groups. n = 4 per group. D–F). ELISA‐based measurement of TNF‐α, IL‐10, and IL‐1β levels in supernatants prepared from the culture medium after each of the different treatments in (B). n = 5 per group. G). Schematic diagram of the experimental schedule for the CAIA mouse model. H). µCT scans showing pathological new bone formation in hind paws in different CAIA model mice groups. I). HE staining of CAIA model mice paws and ankle joints sections showing articular destruction and inflammatory cell infiltration. J). Quantitative analysis of structural parameters of new bone by µCT analysis. n = 5 per group. K–M). Concentration of IL‐17A, TNF‐α, and IL‐6 in the serum of CAIA model mice. N. Therapeutic efficacy of the various substances assessed in terms of changes in the main symptoms of CAIA: arthritis clinical score, paw thickness, paw temperature, and pain‐associated behavior analysis using the von Frey filament test. n = 5 per group. DCFH‐DA, dichlorodihydrofluorescein diacetate; ROS, reactive oxygen species.

Article Snippet: The following ELISA kits were used according to the manufacturer's protocol: CXCL12 (Solarbio, China), TNF‐α (Solarbio, China), IL‐1β (Solarbio, China), IL‐6 (Solarbio, China), and MMP14 (Thermo Scientific, USA).

Techniques: Activation Assay, In Vivo, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Staining, Concentration Assay, Drug discovery