Journal: Molecular medicine reports

Article Title: Time‑order effects of vitamin C on hexavalent chromium‑induced mitochondrial damage and DNA‑protein crosslinks in cultured rat peripheral blood lymphocytes.

doi: 10.3892/mmr.2013.1462

Figure Lengend Snippet: Figure 3. Time‑order effect of vit C on Cr(VI)‑induced mitochondrial damage in PBLs. Different time‑order effects of vit C [pre‑treatment with vit C for 2 h plus Cr(VI); co‑treatment with Cr(VI) and vit C; and Cr(VI) treatment for 2 h plus vit C post‑treatment] on Cr(VI)‑induced mitochon drial damage were detected. Two concentrations of vit C (100 and 200 µM) were employed. (A) MDA content was determined using the MDA Content Detection Assay kit according to the manufacturer's instructions. (B) The PBLs were collected following chemical exposure and were then processed for mitochondrial isolation. The PTP open rate was examined using a fluores cence spectrophotometer. (C) The MMP was monitored by the fluorescence changes in a specific fluorescent cationic dye, Rh123. Data are expressed as the mean ± standard deviation of three independent experiments. vit C, vitamin C; Cr(VI), hexavalent chromium; PBL, peripheral blood lympho cyte; MDA, malondialdehyde; PTP, permeability transition pore; MMP, mitochondrial membrane potential; Rh123, rhodamine 123.

Article Snippet: The mitochondrial ROS production was determined spectrofluorimetrically, by detecting the fluorescence intensity of 2',7'‐dichlorofluorescein (DCF), the oxidized product of the membrane permeable fluoroprobe 5‐(and 6)‐chloromethyl‐2',7'‐dichlorodihydrofluorescein diacetate (CM-H2DCFDA; Molecular Probes, Carlsbad, CA, USA).

Techniques: Detection Assay, Isolation, Spectrophotometry, Fluorescence, Standard Deviation, Permeability, Membrane

Journal: Molecular medicine reports

Article Title: Time‑order effects of vitamin C on hexavalent chromium‑induced mitochondrial damage and DNA‑protein crosslinks in cultured rat peripheral blood lymphocytes.

doi: 10.3892/mmr.2013.1462

Figure Lengend Snippet: Figure 4. Time‑order effect of vit C on Cr(VI)‑induced ROS accumulation in PBLs. (A) Cr(VI) induces ROS accumulation. We measured the ROS levels in PBLs exposed to various concentrations of Cr(VI) (25, 50, 100, 200 and 400 µM) by utilizing the oxidant‑sensitive fluorogenic probe CM‑H2DCFDA. The level of ROS production, which was considered to be directly proportional to fluorescence intensity, was quantitated by flow cytometry (upper panel) and was detected under a fluorescence microscope (bottom panel). *P<0.05 and **P<0.01 compared with the control group. (B) Different time‑order effects of vit C [pre‑treatment with vit C for 2 h plus Cr(VI); co‑treatment with Cr(VI) and vit C; and Cr(VI) treatment for 2 h plus vit C post‑treatment] on Cr(VI)‑induced ROS accumulation were detected. Two concentrations of vit C (100 and 200 µM) were employed. Data are expressed as the mean ± standard deviation of three independent experiments. vit C, vitamin C; Cr(VI), hexavalent chromium; PBL, peripheral blood lymphocyte; ROS, reactive oxygen species; CM‑H2DCFDA, 5‑(and 6)‑chloromethyl‑2',7'‑dichlorodihydrofluorescein diacetate.

Article Snippet: The mitochondrial ROS production was determined spectrofluorimetrically, by detecting the fluorescence intensity of 2',7'‐dichlorofluorescein (DCF), the oxidized product of the membrane permeable fluoroprobe 5‐(and 6)‐chloromethyl‐2',7'‐dichlorodihydrofluorescein diacetate (CM-H2DCFDA; Molecular Probes, Carlsbad, CA, USA).

Techniques: Fluorescence, Flow Cytometry, Microscopy, Control, Standard Deviation

Journal: Journal of Nanobiotechnology

Article Title: AE-MXene-modified titanium alloy promotes osseointegration by regulating the AMPK-MTOR-autophagy pathway in macrophage

doi: 10.1186/s12951-026-04080-3

Figure Lengend Snippet: AE-MXene induced autophagy in macrophages. a ) The basic technical flowchart of Data-Independent Acquisition (DIA)-based quantitative proteomics. b ) Volcano plot of the distinct upregulated and downregulated genes of ML (AE-MXene (0.1 mg/mL) + LPS) versus TL (TA + LPS). c ) Functional enrichment map of differentially expressed proteins in the TL and ML groups. d ) Heatmap of differentially expressed genes associated with autophagy and antioxidative processes of ML versus TL. e ) Expression of autophagy-related genes analyzed by RT-qPCR in RAW264.7 cells in the different groups ( n ≥ 3). ( f ) Western blot images of P62, LC3I/II, and GAPDH in RAW264.7 cells from different treatment groups. The experimental procedure was as follows: RAW264.7 cells were pretreated with CQ (5 µM) or 3-MA (5 mM) as required by experimental conditions for 2 h, followed by stimulation with fresh medium containing LPS (100 ng/mL) for 24 h. ( g ) Quantitative analysis of P62 and LC3-II/LC3-I ratio in RAW264.7 cells based on Western blot images ( n ≥ 3). ( h ) Representative images from autophagic flux visualization assays using the mRFP-GFP-LC3 tandem fluorescence reporter system. Stable RAW264.7 cells expressing this reporter were seeded onto material surfaces. Depending on experimental conditions, cells were either pretreated with CQ (5 µM) or 3-MA (5 mM) for 2 h, followed by stimulation with fresh medium containing LPS (100 ng/mL) for 24 h. Autophagosomes (APs) appear as yellow puncta (mRFP + GFP + ), whereas autophagolysosomes (ALs) exhibit only red fluorescent puncta (mRFP + GFP – ). Scale bar = 20 μm. ( i ) Quantitative statistical analysis of the number of APs, number of ALs, and the AL/AP ratio derived from fluorescence images ( n ≥ 3). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001

Article Snippet: Intracellular ROS Detection: The generation of reactive oxygen species (ROS) was measured using a fluorescent probe (DCFH-DA, Beyotime).

Techniques: Data-independent acquisition, Quantitative Proteomics, Functional Assay, Expressing, Quantitative RT-PCR, Western Blot, Fluorescence, Derivative Assay

Journal: Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy

Article Title:

Gross saponin of Tribulus terrestris improves erectile dysfunction in type 2 diabetic rats by repairing the endothelial function of the penile corpus cavernosum

doi: 10.2147/dmso.s205722

Figure Lengend Snippet: Figure 9 Comparison of apoptotic index in the corpus cavernosum of each group. Note: (A): Representative TUNEL staining map, positive apoptotic cell nucleus is fluorescent green; (B): difference in apoptosis index between different treatment groups; *P<0.05. Abbreviations: T2DMED, type 2 diabetes mellitus; GSTT, gross saponins of Tribulus terrestrise.

Article Snippet: Determination of reactive oxygen species (ROS) levels As per Davidson et al,14 the expression of ROS in each group was detected using a fluorescent probe with DHE superoxide anion level (Servicebio, Wuhan, China).

Techniques: Comparison, TUNEL Assay, Staining