Journal: Endocrinology and Metabolism

Article Title: Fancd2os Reduces Testosterone Production by Inhibiting Steroidogenic Enzymes and Promoting Cellular Apoptosis in Murine Testicular Leydig Cells

doi: 10.3803/enm.2022.1431

Figure Lengend Snippet: Fig. 4. Fancd2 opposite-strand (Fancd2os) reduces testosterone production and steroidogenic enzyme expression in TM3 cells. The testoster one levels in Fancd2os-overexpressing (A) or knockdown TM3 cells (B) were detected by enzyme-linked immunosorbent assays (n=3 per group). (C, D, E) Relative quantities of mRNA expression of steroidogenic acute regulatory protein (StAR), P450 cholesterol side-chain cleavage (P450scc), and 3β-hydroxysteroid dehydrogenase (3β-HSD) in Fancd2os-overexpressing TM3 cells or Fancd2os knockdown TM3 cells (F, G, H) were determined real-time polymerase chain reaction using β-actin as a housekeeping gene. Each bar represents the mean± standard deviation from three separate experiments. Significant difference compared to TM3, vector/TM3 or NC/TM3. aP<0.05; bP<0.01.

Article Snippet: The testosterone concentration of both serum and cell supernatants was measured using a testosterone ELISA Kit (Elabscience, Houston, TX, USA) according to the manufacturer’s protocol.

Techniques: Expressing, Knockdown, Real-time Polymerase Chain Reaction, Standard Deviation, Plasmid Preparation

Journal: Endocrinology and Metabolism

Article Title: Fancd2os Reduces Testosterone Production by Inhibiting Steroidogenic Enzymes and Promoting Cellular Apoptosis in Murine Testicular Leydig Cells

doi: 10.3803/enm.2022.1431

Figure Lengend Snippet: Fig. 6. Higher Fancd2 opposite-strand (Fancd2os) levels in older mouse Leydig cells result in cellular apoptosis and lower serum testosterone production. (A) The serum testosterone levels from mice of different ages were measured using enzyme-linked immunosorbent assays. (B, C) The testis tissues from different aged mice were sliced and then Fancd2os protein expression and apoptosis were analyzed using immuno chemistry and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay, respectively. ST represents seminiferous tubule. Black arrows and red arrows indicate Fancd2os-positive cells and TUNEL-positive cells, respectively. Scale bar repre sents 25 μm. The results are given as the mean±standard deviation (n=3). (D) The correlations between Fancd2os expression (B) and the TUNEL-positive staining rate (C) were analyzed using Pearson correlation coefficients. An r≥0.5 was considered to indicate a strong correla tion, and P<0.05 was considered statistically significant. aP<0.05 compared with juvenile mice; bP<0.05 compared with young mice; cP<0.05 compared with middle-aged mice.

Article Snippet: The testosterone concentration of both serum and cell supernatants was measured using a testosterone ELISA Kit (Elabscience, Houston, TX, USA) according to the manufacturer’s protocol.

Techniques: Expressing, End Labeling, TUNEL Assay, Standard Deviation, Staining

Journal: Ecotoxicology and environmental safety

Article Title: Ameliorative effect of betulinic acid against zearalenone exposure triggers testicular dysfunction and oxidative stress in mice via p38/ERK MAPK inhibition and Nrf2-mediated antioxidant defense activation.

doi: 10.1016/j.ecoenv.2022.113561

Figure Lengend Snippet: Fig. 2. BA protected the aggravation of male reproduction injury in ZEA-induced mice. The morphology of the sperm was photographed using an optical microscope (A). The sperm motility was used to evaluate the male repro duction of mice, including sperm survival rate (B), sperm malformation rate (C), and sperm mortality rate (D). Protein and mRNA levels of ERα in testis were detected by immunoblotting and RT-PCR analysis, respectively, and the pro tein and mRNA levels were normalized to β-actin (E-G). The content of testosterone in serum was detected by ELISA kit (H). The mRNA expression of CLDN11 (I), CDH2 (J), and Vim (K) were measured by RT-PCR. Mean ± SEM, *P < 0.05 and **P < 0.01 represented a significant differ ence compared to the control group, while #P < 0.05 and ##P < 0.01 represented a signifi cant difference compared to the ZEA group.

Article Snippet: Testosterone enzyme linked immunosorbent assay (ELISA) kit (CSB-E05101m) was obtained from Cusabio Biotech Co. Ltd. (Wuhan, China).

Techniques: Microscopy, Western Blot, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Expressing, Control

Journal: Egyptian Pediatric Association Gazette

Article Title: Cardiac troponin T in neonates with respiratory distress

doi: 10.1016/j.epag.2018.09.004

Figure Lengend Snippet: Fig. 1. Cardiac troponin T in patients with and without inotropic support.

Article Snippet: The assay applies the quantitative sandwich enzyme immunoassay technique using Human cardiac troponin T(cTnT) ELISA Kit (CUSABIO BIOTECH CO., Ltd., China).

Techniques:

Journal: Egyptian Pediatric Association Gazette

Article Title: Cardiac troponin T in neonates with respiratory distress

doi: 10.1016/j.epag.2018.09.004

Figure Lengend Snippet: Fig. 2. Cardiac troponin T in survivors and non-survivors.

Article Snippet: The assay applies the quantitative sandwich enzyme immunoassay technique using Human cardiac troponin T(cTnT) ELISA Kit (CUSABIO BIOTECH CO., Ltd., China).

Techniques:

Journal: The Journal of Immunology Author Choice

Article Title: Macrophagic Extracellular Vesicle CXCL2 Recruits and Activates the Neutrophil CXCR2/PKC/NOX4 Axis in Sepsis

doi: 10.4049/jimmunol.2100229

Figure Lengend Snippet: Chemokine profile of the control and LPS-induced Raw264.7 EVs. (A) The chemokine profile of EVs from control and LPS-induced Raw264.7 cells. (B) The CXCL2 level (mean ± SD) in the control-Raw-EV and LPS-Raw-EV groups. The experiments were repeated three times. **p < 0.01 versus the control-Raw264.7-EV group. (C) PKH67-labeled control-Raw264.7-EVs and LPS-Raw264.7-EVs were injected into wild-type C57BL/6 mice (n = 6 per group) via the tail vein. The colocalization of PKH67-EVs and CXCL2 in the liver was detected by immunofluorescence (original magnification × 200). (D and E) GO analysis and KEGG analysis of various chemokines expressed in EVs from control and LPS-induced Raw264.7 cells.

Article Snippet: The quantification of CXCL2 (KE10022; Proteintech), CXCL7 (DY393; R&D Systems, Minneapolis, MN), CXCL17 (AD7670Hu; Andy Gene, Beijing, China), CCL2 (KE00091; Proteintech), CCL3 (KE00092; Proteintech), and CCL5 (KE00093; Proteintech) was performed using quantitative ELISA.

Techniques: Control, Labeling, Injection, Immunofluorescence

Journal: The Journal of Immunology Author Choice

Article Title: Macrophagic Extracellular Vesicle CXCL2 Recruits and Activates the Neutrophil CXCR2/PKC/NOX4 Axis in Sepsis

doi: 10.4049/jimmunol.2100229

Figure Lengend Snippet: LPS-induced CXCL2 of Raw264.7 EVs attracts neutrophils in vitro and in vivo and activates neutrophils via the CXCR2/PKC/NOX4 pathway. (A) The CXCL2 level (mean ± SD) of Raw264.7 in the control, LPS, LPS + lenti-CXCL2, and LPS + siRNA-CXCL2 groups detected by Western blotting (WB). The experiments were repeated three times. *p < 0.01 versus the control group, &p < 0.01 versus the LPS group, #p < 0.01 versus the LPS group. (B) The CXCL2 level (mean ± SD) in Raw264.7 EVs of the control, LPS, LPS + lenti-CXCL2, and LPS + siRNA-CXCL2 groups detected by WB. The experiments were repeated three times. (C) Raw264.7 EVs derived from the control, LPS, LPS + lenti-CXCL2, and LPS + siRNA-CXCL2 groups induced BMDN chemotaxis (mean ± SD) (n = 5 fields per group). (D) The expression of CXCR2, PKC, and NOX4 (mean ± SD) of BMDNs in the control EVs, LPS-EVs, (LPS + lenti-CXCL2)-EVs, and (LPS + siRNA-CXCL2)-EVs groups was detected by WB. The experiments were repeated three times. (E) H&E staining, MPO+ cells, and Ly-6G+ cells (mean ± SD) in the liver of the control EVs, LPS-EVs, (LPS + lenti-CXCL2)-EVs, and (LPS + siRNA-CXCL2)-EV group mice (n = 6 per group) detected by immunohistochemistry. Scale bars, 100 μm. (F–H) ALT, AST, and LDH levels (mean ± SD) in the serum of the control EVs, LPS-EVs, (LPS + lenti-CXCL2)-EVs, and (LPS + siRNA-CXCL2)-EV group mice (n = 6 per group). (I) The number of systemic circulating neutrophils (mean ± SD) in wild-type C57BL/6 mice treated with control EVs, LPS-EVs, (LPS + lenti-CXCL2)-EVs, and (LPS + siRNA-CXCL2)-EVs (n = 3 per group). (J) The expression of CXCR2, PKC, and NOX4 (mean ± SD) in the liver of the control EVs, LPS-EVs, (LPS + lenti-CXCL2)-EVs, and (LPS + siRNA-CXCL2)-EV group mice was detected by WB. The experiments were repeated three times. *p < 0.01 versus the control-Raw264.7-EV group, &p < 0.01 versus the LPS-Raw264.7-EV group, #p < 0.01 versus the LPS-Raw264.7-EV group.

Article Snippet: The quantification of CXCL2 (KE10022; Proteintech), CXCL7 (DY393; R&D Systems, Minneapolis, MN), CXCL17 (AD7670Hu; Andy Gene, Beijing, China), CCL2 (KE00091; Proteintech), CCL3 (KE00092; Proteintech), and CCL5 (KE00093; Proteintech) was performed using quantitative ELISA.

Techniques: In Vitro, In Vivo, Control, Western Blot, Derivative Assay, Chemotaxis Assay, Expressing, Staining, Immunohistochemistry

Journal: The Journal of Immunology Author Choice

Article Title: Macrophagic Extracellular Vesicle CXCL2 Recruits and Activates the Neutrophil CXCR2/PKC/NOX4 Axis in Sepsis

doi: 10.4049/jimmunol.2100229

Figure Lengend Snippet: EVs from the serum of patients with sepsis attract neutrophils and activate the neutrophil CXCR2/PKC/NOX4 pathway in vitro. (A) The chemokine profile of EVs from the serum of healthy individuals and patients with sepsis. (B–G) CXCL2, CXCL7, CXCL17, CCL2, CCL3, and CCL5 levels (mean ± SD) in EVs from the serum of healthy individuals (n = 8) and patients with sepsis (n = 26) detected using ELISA. *p < 0.05 versus the healthy-EV group, ***p < 0.01 versus the healthy-EV group. (H) sEVs derived from healthy control (HC) and patients with sepsis-induced PMN chemotaxis (mean ± SD) (n = 5 fields per group). (I) The expression of CXCR2, PKC, and NOX4 (mean ± SD) in the PMNs of the HC-sEV– and sepsis-sEV–treated groups was detected by Western blotting. The experiments were repeated three times. **p < 0.01 versus the healthy-EV group, #p < 0.05 versus the healthy-EV group, &p < 0.05 versus the healthy-EV group.

Article Snippet: The quantification of CXCL2 (KE10022; Proteintech), CXCL7 (DY393; R&D Systems, Minneapolis, MN), CXCL17 (AD7670Hu; Andy Gene, Beijing, China), CCL2 (KE00091; Proteintech), CCL3 (KE00092; Proteintech), and CCL5 (KE00093; Proteintech) was performed using quantitative ELISA.

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Derivative Assay, Control, Chemotaxis Assay, Expressing, Western Blot

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Cancer-associated fibroblasts promote pro-tumor functions of neutrophils in pancreatic cancer via IL-8: potential suppression by pirfenidone

doi: 10.1007/s00262-025-03946-z

Figure Lengend Snippet: IL-8 secreted by activated neutrophils is suppressed by PFD. A IL-8 secretion by neutrophils measured by ELISA. IL-8 secretion was significantly increased in neutrophils stimulated with CAF-CM compared to non-stimulated neutrophils. PFD administration during neutrophil activation with CAF-CM significantly suppressed IL-8 secretion. B Effect of secretions from activated neutrophils on cancer cell proliferation. The presence of neutrophils significantly increased MIA PaCa-2 proliferation. Mean ± SD is shown. Each dot represents the number of cells per well. Student’s t-test was used. ***, P < 0.001. C Pancreatic cancer cells induce fibroblasts to become iCAFs, and iCAF-CM activates neutrophils, increasing their IL-8 secretion and enhancing the malignant traits of pancreatic cancer cells. Conversely, PFD acts on activated neutrophils, suppressing the malignant traits of pancreatic cancer cells. This figure was created with BioRender.com

Article Snippet: To evaluate IL-8 levels in neutrophil secretions, culture supernatants were assayed using a human IL-8 enzyme-linked immunosorbent assay (ELISA) kit (Proteintech, KE00006) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Activation Assay

Journal: Journal of Thrombosis and Haemostasis

Article Title: Non‐severe COVID‐19 is associated with endothelial damage and hypercoagulability despite pharmacological thromboprophylaxis

doi: 10.1111/jth.15660

Figure Lengend Snippet: Plasma tissue plasminogen activator (t‐PA) levels are significantly elevated in coronavirus disease 2019 (COVID‐19). Plasma levels of t‐PA (A) were significantly elevated in hospitalized patients with non‐severe COVID‐19 ( n = 14) compared to a group of hospitalized patients ( n = 11) who had screened negative for severe acute respiratory syndrome coronavirus 2 by real‐time polymerase chain reaction. Levels of soluble thrombomodulin (sTM; B), vascular cell adhesion molecule‐1 (VCAM‐1; C), intercellular adhesion molecule‐1 (ICAM‐1; D), plasminogen activator inhibitor‐1 (PAI‐1; E) and E‐selectin (F) were similar in both groups. All samples were assayed in technical duplicate. Data are presented as the mean ± standard deviation. Statistical analysis was performed using a two‐tailed Student’s t test. * P < .05

Article Snippet: ELISAs for TFPI (DTFP10), tissue plasminogen activator (t‐PA; DTPA00), thrombomodulin (DTHBD0), vascular cell adhesion molecule‐1 (VCAM‐1; DVC00), intercellular adhesion molecule‐1 (ICAM‐1; DCIM00), and E‐selectin (DSLE00) were purchased from R&D Systems and a plasminogen activator inhibitor‐1 (PAI‐1; ab108891) ELISA was purchased from Abcam.

Techniques: Clinical Proteomics, Real-time Polymerase Chain Reaction, Standard Deviation, Two Tailed Test