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3 protocols using levofloxacin lev

1

Anti-inflammatory activity of natural compounds

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ATG, SPA, CLA, CGA, arctiin, deoxycholic acid, glycocholic acid and deoxyglycocholic acid were obtained from Yifang S&T (Tianjin, China). Enzyme-linked immunosorbent assay (ELISA) assay kits of rat TNF-α, IL-6, IL-1β and RANTES, as well as human intracellular proteins (total p38, JNK and ERK), were obtained from Pierce/Endogen Co. (Rockford, Illinois, USA). Human TNF-α was purchased from PeproTech (Rocky Hill, NJ, USA). Dexamethasone (Dex) and levofloxacin (LEV) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Reagents for cell culture were obtained from Gibco BRL Life Technologies (Rockville, MD, USA). The NF-κB luciferase reporter plasmid pGL4.32 and Renilla luciferase reporter vector plasmid pRL-TK were obtained from Promega Co. (Fitchburg, WI, USA). The Lipofectamine 2000 transfection reagent was obtained from Invitrogen (Carlsbad, CA, USA). The anti-IκB-α antibody was obtained from Santa Cruz Biotechnology (San Diego, CA, USA). The P. aeruginosa PAK strain, a clinical isolate from the sputum of a patient suffering from bronchiectasis, was provided by Professor Mingqiang Qiao (College of Life Science, Nankai University, China). All other reagents used in this study were of analytical grade.
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2

Synthesis of Cyclodextrin-Based Polymer Carriers

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β-cyclodextrin (CD—Cyclolab, Budapest, Hungary) was dried under vacuum (0.01 mbar at 80 °C for 72 h) and kept under an argon atmosphere. ε-Caprolactone (CL—Sigma-Aldrich, Saint Louis, MO, USA), N,N-Dimethylformamide (DMF—Sigma-Aldrich, Saint Louis, MO, USA), and dimethyl sulfoxide (DMSO—Sigma-Aldrich, Saint Louis, MO, USA) were distilled under reduced pressure before use. Polyethylene glycol (PEG, Mn = 2000 g/mol—Sigma Aldrich, Saint Louis, MO, USA) was dried under reduced pressure for 3 h, at 100 °C to remove traces of water. Isophorone diisocyanate (IPDI), dibutyltin dilaurate (DBTL), 4-dimethylaminopyridine (DMAP), Amberlyst 15 hydrogen, and levofloxacin (LEV) were purchased from Sigma-Aldrich, Saint Louis, MO, USA, and were used as received.
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3

Chitosan-based Antimicrobial Nano-Formulations

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Chitosan (CS, low molecular weight, 88% hydrolysis degree, Sigma Aldrich, Saint Louis, MO, USA); Poly(ethylene glycol) methyl ether acrylate (PEGA, Mn = 480, Sigma Aldrich, Saint Louis, MO, USA); acetic acid (CH3COOH, 99%, Sigma Aldrich, Saint Louis, MO, USA); glutaraldehyde (GA, Sigma Aldrich, 25% aqueous solution, Saint Louis, MO, USA); sodium sulfate (Na2SO4, Sigma Aldrich, Saint Louis, MO, USA); levofloxacin (LEV, Sigma Aldrich, Saint Louis, MO, USA); ciprofloxacin (CP, Sigma Aldrich, Saint Louis, MO, USA); Tween 80 (Sigma Aldrich, Saint Louis, MO, USA); Span 80 (Sigma Aldrich, Saint Louis, MO, USA); acetate and phosphate-buffered saline (ABS and PBS) were purchased from Sigma Aldrich, Saint Louis, MO, USA; MCF-10A cells were purchased from American Type Culture Collection, Manassas, VA, USA; DMEM/F12 supplemented with 5% horse serum (Sigma Aldrich, Saint Louis, MO, USA); 20 ng/mL EGF (Sigma Aldrich, Saint Louis, MO, USA); HDMVEC (Primary Dermal Microvascular Endothelial Cells) are from ATCC, Manassas, VA, USA; Milli-Q ultrapure distilled water (Merck, Rahway, NJ, USA). All other reagents used in this study were of analytical grade purity and were used without further purification. The human blood samples used were freshly obtained from one healthy nonsmoker volunteer.
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