Journal: Balkan Medical Journal

Article Title: Drug-eluting Vein Graft with Acetylsalicylic Acid-Ticagrelor-Unfractionated Heparin Complex Inhibits Early Graft Thrombosis

doi: 10.4274/balkanmedj.galenos.2020.2020.1.128

Figure Lengend Snippet: Von Willebrand factor (VWF) immunohistochemical staining of the study groups on the 1 st and 3 rd days. It was observed that VWF staining was lower in the locally acting drug group compared with that in the untreated and pluronic gel groups. From a general point of view, the lowest staining was observed with preparations containing acetylsalicylic acid + ticagrelor + unfractionated heparin (magnification 200x).

Article Snippet: Immunohistochemical staining was performed with the anti-rat von Willebrand factor (VWF) polyclonal antibody kit (Bioss, USA, lot no: AD071046) following the manufacturer’s instructions.

Techniques: Immunohistochemical staining, Staining

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Growth Differentiation Factor 7 Prevents Sepsis-Induced Acute Lung Injury in Mice

doi: 10.1155/2022/3676444

Figure Lengend Snippet: GDF7 alleviates LPS-induced ALI and pulmonary dysfunction in mice. (a) Mice were intratracheally injected with LPS (5 mg/kg in 50 μ L saline) to generate sepsis-induced ALI, and serum GDF7 levels were detected using an ELISA kit after 12 h ( n = 6). (b) The levels of Gdf7 mRNA in lung tissues with or without LPS instillation were detected using quantitative real-time PCR ( n = 6). (c) The levels of GDF7 protein in lung tissues with or without LPS instillation were detected using western blot ( n = 6). (d-e) Mice were subcutaneously injected with rmGDF7 (25 μ g per mouse), and serum and lung GDF7 levels in mice were detected using an ELISA kit after 24 h ( n = 6). (f) Mice were subcutaneously injected with rmGDF7 (25 μ g per mouse), and 24 h later, ALI mice were intratracheally injected with LPS (5 mg/kg in 50 μ L saline) to generate sepsis-induced ALI. After 12 h, fresh lungs were harvested for the analysis of LDH activity using a commercial kit ( n = 6). (g) Lung wet to dry ratio ( n = 6). (h) Total proteins in BALF ( n = 6). (i) Arterial blood gas analysis results ( n = 6). (j) Respiratory function was detected by Buxco, including pulmonary ventilation, lung compliance, and airway resistance ( n = 6). (k) Mice were subcutaneously injected with rmGDF7 (25 μ g per mouse), and 24 h later, ALI mice were intratracheally injected with a lethal dose of LPS (25 mg/kg). The survival rate was monitored every 12 h post-LPS treatment ( n = 20). All data were presented as mean ± SD, and ∗ P < 0.05 was regarded to be statistically significant.

Article Snippet: Lactate Dehydrogenase (LDH) Assay Kit (#ab102526), Mouse Myeloperoxidase (MPO) ELISA Kit (#ab155458), Lipid Peroxidation (MDA) Assay Kit (#ab118970), Lipid Peroxidation (4-HNE) Assay Kit (#ab238538), Superoxide Dismutase (SOD) Activity Assay Kit (#ab65354), and Glutathione (GSH) Assay Kit (#ab239727) were purchased from Abcam (Cambridge, UK).

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Western Blot, Activity Assay

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Growth Differentiation Factor 7 Prevents Sepsis-Induced Acute Lung Injury in Mice

doi: 10.1155/2022/3676444

Figure Lengend Snippet: GDF7 attenuates LPS-induced ALI through activating AMPK in vivo. (a) Mice were subcutaneously injected with rmGDF7 (25 μ g per mouse), and 24 h later, ALI mice were intratracheally injected with LPS (5 mg/kg in 50 μ L saline) to generate sepsis-induced ALI. After 12 h, lung tissues were harvested for the analysis of the AMPK pathway using western blot ( n = 6). (b) To inhibit AMPK, ALI mice were intraperitoneally injected with CpC (20 mg/kg) at 2 h pre- and 2 h post-rmGDF7 injection, and then IL-6 and TNF- α levels in lung tissues were detected ( n = 6). (c) ROS content in lung tissues ( n = 6). (d) MDA and 4-HNE levels in lung tissues ( n = 6). (e) LDH activity in lung tissues ( n = 6). (f) Lung wet to dry ratio ( n = 6). (g) Arterial blood gas analysis results ( n = 6). (h) Respiratory function was detected by Buxco, including pulmonary ventilation, lung compliance, and airway resistance ( n = 6). All data were presented as mean ± SD, and ∗ P < 0.05 was regarded to be statistically significant.

Article Snippet: Lactate Dehydrogenase (LDH) Assay Kit (#ab102526), Mouse Myeloperoxidase (MPO) ELISA Kit (#ab155458), Lipid Peroxidation (MDA) Assay Kit (#ab118970), Lipid Peroxidation (4-HNE) Assay Kit (#ab238538), Superoxide Dismutase (SOD) Activity Assay Kit (#ab65354), and Glutathione (GSH) Assay Kit (#ab239727) were purchased from Abcam (Cambridge, UK).

Techniques: In Vivo, Injection, Western Blot, Activity Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Glutathione peroxidase 4 restrains temporomandibular joint osteoarthritis progression by inhibiting ferroptosis

doi: 10.1111/jcmm.18377

Figure Lengend Snippet: Ferroptosis was activated in TMJOA. (A) 8‐OHdG level in synovial fluid from TMJOA patients and normal patients (patients with temporomandibular fractures). (B) Representative histopathological haematoxylin–eosin, safranin O/fast green and alcian blue staining of undamaged and TMJ tissues from TMJOA patients. (C) Fe 2+ , Fe 3+ , and total iron levels were analysed in synovial tissues from TMJOA and normal patients. (D) Glutathione peroxidase activity levels were detected in synovial tissues from TMJOA and normal patients. (E) GSH contents and ratio of GSH/GSSG were measured in synovial tissues from TMJOA and normal patients. (F) The mRNA level of GPX4 was determined by qRT‐PCR assay in synovial tissues from TMJOA and normal patients. (G) The protein level of GPX4 was determined by western blot assay in synovial tissues from TMJOA and normal patients. ** p < 0.01; *** p < 0.001.

Article Snippet: The levels of Glutathione peroxidase, GSH/GSSG and MDA in the supernatant of lysed cells were assessed with a rat 8‐OHdG ELISA kit (ml028318, MLBIO, Shanghai, China), Glutathione peroxidase ELISA kit (S0056, Beyotime Biotechnology, Shanghai, China), GSH/GSSG ELISA kit (S0053, Beyotime Biotechnology, Shanghai, China) and MDA ELISA kit (S0131S, Beyotime Biotechnology, Shanghai, China) as instructions indicated.

Techniques: Staining, Activity Assay, Quantitative RT-PCR, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Glutathione peroxidase 4 restrains temporomandibular joint osteoarthritis progression by inhibiting ferroptosis

doi: 10.1111/jcmm.18377

Figure Lengend Snippet: Ferroptosis was activated in the primary OA‐FLSs. (A) Fe 2+ , Fe 3+ , and total iron levels in N‐FLSs and OA‐FLSs. (B) Glutathione peroxidase activity levels were detected in N‐FLSs and OA‐FLSs. (C) GSH contents and ratio of GSH/GSSG were measured in N‐FLSs and OA‐FLSs. ** p < 0.01.

Article Snippet: The levels of Glutathione peroxidase, GSH/GSSG and MDA in the supernatant of lysed cells were assessed with a rat 8‐OHdG ELISA kit (ml028318, MLBIO, Shanghai, China), Glutathione peroxidase ELISA kit (S0056, Beyotime Biotechnology, Shanghai, China), GSH/GSSG ELISA kit (S0053, Beyotime Biotechnology, Shanghai, China) and MDA ELISA kit (S0131S, Beyotime Biotechnology, Shanghai, China) as instructions indicated.

Techniques: Activity Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Glutathione peroxidase 4 restrains temporomandibular joint osteoarthritis progression by inhibiting ferroptosis

doi: 10.1111/jcmm.18377

Figure Lengend Snippet: Ferroptosis inhibitors promote HFLSs survival by suppressing ferroptosis. (A) Cell viability was assessed by CCK‐8 assay in indicated groups. (B) The concentration of MDA was assessed by ELISA assay in indicated groups. (C) Glutathione peroxidase activity levels were detected in indicated groups. (D) GSH contents and ratio of GSH/GSSG were measured in indicated groups. (E) The protein levels of GPX4, ADAMTS5, SLC3A2, ACSL4, and GAPDH in indicated groups were determined by western blot assay. ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: The levels of Glutathione peroxidase, GSH/GSSG and MDA in the supernatant of lysed cells were assessed with a rat 8‐OHdG ELISA kit (ml028318, MLBIO, Shanghai, China), Glutathione peroxidase ELISA kit (S0056, Beyotime Biotechnology, Shanghai, China), GSH/GSSG ELISA kit (S0053, Beyotime Biotechnology, Shanghai, China) and MDA ELISA kit (S0131S, Beyotime Biotechnology, Shanghai, China) as instructions indicated.

Techniques: CCK-8 Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Glutathione peroxidase 4 restrains temporomandibular joint osteoarthritis progression by inhibiting ferroptosis

doi: 10.1111/jcmm.18377

Figure Lengend Snippet: GPX4 knockdown inhibited HFLSs survival by activating ferroptosis. (A) The mRNA level of GPX4 in indicated groups was determined by qRT‐PCR assay. (B) The protein levels of GPX4, ADAMTS5, SLC3A2, ACSL4, and GAPDH in indicated groups were determined by western blot assay. (C) Cell viability was assessed by CCK‐8 assay in indicated groups. (D) Fe 2+ , Fe 3+ , and total iron level in indicated groups. (E) Glutathione peroxidase activity levels were detected in indicated groups. (F) GSH contents and ratio of GSH/GSSG were measured in indicated groups. (G) The concentration of MDA was assessed by ELISA assay in indicated groups. ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: The levels of Glutathione peroxidase, GSH/GSSG and MDA in the supernatant of lysed cells were assessed with a rat 8‐OHdG ELISA kit (ml028318, MLBIO, Shanghai, China), Glutathione peroxidase ELISA kit (S0056, Beyotime Biotechnology, Shanghai, China), GSH/GSSG ELISA kit (S0053, Beyotime Biotechnology, Shanghai, China) and MDA ELISA kit (S0131S, Beyotime Biotechnology, Shanghai, China) as instructions indicated.

Techniques: Quantitative RT-PCR, Western Blot, CCK-8 Assay, Activity Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Glutathione peroxidase 4 restrains temporomandibular joint osteoarthritis progression by inhibiting ferroptosis

doi: 10.1111/jcmm.18377

Figure Lengend Snippet: GPX4 overexpression promoted HFLSs survival by inhibiting ferroptosis. (A) The protein levels of GPX4, ADAMTS5, SLC3A2, ACSL4, and GAPDH in indicated groups were determined by western blot assay. (B) Cell viability was assessed by CCK‐8 assay in indicated groups. (C) Fe 2+ , Fe 3+ , and total iron level in indicated groups. (D) Glutathione peroxidase activity levels were detected in indicated groups. (E) GSH contents and ratio of GSH/GSSG were measured in indicated groups. (F) The concentration of MDA was assessed by ELISA assay in indicated groups. ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: The levels of Glutathione peroxidase, GSH/GSSG and MDA in the supernatant of lysed cells were assessed with a rat 8‐OHdG ELISA kit (ml028318, MLBIO, Shanghai, China), Glutathione peroxidase ELISA kit (S0056, Beyotime Biotechnology, Shanghai, China), GSH/GSSG ELISA kit (S0053, Beyotime Biotechnology, Shanghai, China) and MDA ELISA kit (S0131S, Beyotime Biotechnology, Shanghai, China) as instructions indicated.

Techniques: Over Expression, Western Blot, CCK-8 Assay, Activity Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Glutathione peroxidase 4 restrains temporomandibular joint osteoarthritis progression by inhibiting ferroptosis

doi: 10.1111/jcmm.18377

Figure Lengend Snippet: GPX4 Overexpression and Fer‐1 significantly improve the pathological changes in the TMJOA Rat Model. (A) The analgesic effect of celecoxib on chronic inflammation in rats was evaluated using the hot water tail‐flick test (HWT). (B) HE staining and Safranin O staining depicts the changes in cartilage structure and matrix proteoglycans of TMJ tissues. (C) Severity analysis of the TMJOA model using the OARSI scoring system. (D) Quantification of chondrocyte counts in the histological slices. (E) Statistical analysis of each group's percentage of proteoglycan area. (F–H) Assessment of iron concentration in various groups using an iron assay kit in TMJ synovial tissues. (I, J) The measurement of GSH contents and the ratio of GSH/GSSG in TMJ synovial tissues. (K, L) Immunohistochemistry (IHC) staining and evaluation of GPX4 expression intensity in TMJ tissues. Compared with Sham group, *** p < 0.001; Compared with TMJOA group, ## p < 0.01.

Article Snippet: The levels of Glutathione peroxidase, GSH/GSSG and MDA in the supernatant of lysed cells were assessed with a rat 8‐OHdG ELISA kit (ml028318, MLBIO, Shanghai, China), Glutathione peroxidase ELISA kit (S0056, Beyotime Biotechnology, Shanghai, China), GSH/GSSG ELISA kit (S0053, Beyotime Biotechnology, Shanghai, China) and MDA ELISA kit (S0131S, Beyotime Biotechnology, Shanghai, China) as instructions indicated.

Techniques: Over Expression, Tail Flick Test, Staining, Concentration Assay, Iron Assay, Immunohistochemistry, Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: Aldh2 gene reduces oxidative stress in the bladder by regulating the NF-κB pathway in a mouse model of ketamine-induced cystitis

doi: 10.3892/etm.2020.9239

Figure Lengend Snippet: Ketamine increases the level of oxidative stress in aldh2 KO mice and aggravates pathological damage. (A) Effects of ketamine on parameters of oxidative stress in WT and KO mice at 4, 8 and 12 weeks as detected by ELISA. Negative control mice were treated with NS. * P<0.05 and ** P<0.01. KO, knock-out; NS, normal saline; KHK, knock-out high-dose ketamine group; SOD, superoxide dismutase; GSH, glutathione-sulfhydryl; MDA, malondialdehyde; WHK, wild-type high-dose ketamine group; COX-2, cyclooxygenase 2; iNOS, inducible nitric oxide synthase; WT, wild-type; KNS, knock-out normal saline control group; WNS, wild-type normal saline control group; KLK, knock-out low-dose ketamine group; WLK, wild-type low-dose ketamine group; HK, high-dose ketamine; W, week; -, knock-out; +, wild-type. Ketamine increases the level of oxidative stress in aldh2 KO mice and aggravates pathological damage. (B) Representative hematoxylin and eosin staining images of bladder tissues from KO and WT mice in week 12. Magnification, x100 for the upper images; x400 for the lower images. (C) Representative immunohistochemical staining images of COX-2 and iNOS proteins in the bladder tissues of KO and WT mice in weeks 4 and 12. The cytoplasm and cell membranes exhibiting brown-yellow colors were considered as positive expression of the target protein, which were mainly confined to the bladder epithelium Magnification, x200. (D) Representative Masson trichrome staining images of bladder tissues of KO and WT mice in week 12. Collagen fibers stained green, muscle fibers stained red, and nucleus stained blue-brown. Magnification, x200. Data are presented as the mean ± standard error of the mean from ≥3 experimental repeats. * P<0.05 and ** P<0.01. KO, knock-out; NS, normal saline; KHK, knock-out high-dose ketamine group; SOD, superoxide dismutase; GSH, glutathione-sulfhydryl; MDA, malondialdehyde; WHK, wild-type high-dose ketamine group; COX-2, cyclooxygenase 2; iNOS, inducible nitric oxide synthase; WT, wild-type; KNS, knock-out normal saline control group; WNS, wild-type normal saline control group; KLK, knock-out low-dose ketamine group; WLK, wild-type low-dose ketamine group; HK, high-dose ketamine; W, week; -, knock-out; +, wild-type.

Article Snippet: Next, the solution was centrifuged at 20,000 rpm (41,800 x g) for 10 min at 4 ° C. The supernatant was used to estimate the levels of oxidative stress indicators SOD, GSH and MDA using their respective ELISA kits (SOD, cat. no. 706002; GSH, cat. no. 703002; MDA, grant no. 700870; Cayman Chemical Company), according to the manufacturer's protocols.

Techniques: Enzyme-linked Immunosorbent Assay, Negative Control, Knock-Out, Saline, Control, Staining, Immunohistochemical staining, Expressing