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6 protocols using magnolol

1

Diclazuril and Magnolol Anticoccidial Protocol

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Diclazuril (20201107), a commercial chemical anticoccidial drug, was purchased from Ruipu Biotechnology Co., Ltd. (Tianjin, China), and Magnolol (≥98%, CAS: 528-43-8) was purchased from Yuanye Biotechnology Co. Ltd. (Shanghai, China).
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2

Investigating Anti-Inflammatory Effects of TCM Formulation

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Dexamethasone was obtained from Beijing Solarbia Technology Co., Ltd. (Beijing, China). IL-6, IL-1β, TNF-α enzyme-linked immunosorbent assay (ELISA) and arginase-1 (Arg-1) ELISA kit was obtained from Shanghai Enzymatic Union Biotechnology Co., Ltd. (Shanghai, China). Inducible nitric oxide synthase (iNOS) and bicinchoninic acid (BCA) protein concentration assay kits were purchased from Nanjing Jiancheng Biological Engineering Institute (Nanjing, China). CD86 polyclonal antibody and CD206 polyclonal antibody were purchased from Proteintech Group, Inc. (Wuhang, China). Poly(I:C) and reference standards of chlorogenic acid, forsythoside A, liquiritin, saikosaponin A, baicalin, arginine, magnolol, protocatechuic acid, 4-hydroxyacetophenone, aristolochic acid A and artemisinin were obtained from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). All herbs used to prepare QWJR were obtained from Sinopharm Yunnan, Ltd. (Kunming, China).
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3

Analytical Method Development for Herbal Compounds

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Acetonitrile (ACN, LC-MS/MS grade) and formic acid (≥98%, analytical grade) were purchased from Fisher Scientific (Fair Lawn, New Jersey, USA) and Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China), respectively. Purified water was obtained from Millipore using a Milli-Q system (Bedford, MA). The reference standards (emodin, rheic acid, chrysophanic acid, magnolol, honokiol, hesperidin, naringin and neohesperidin) were purchased from Shanghai Yuanye Biotechnology Co., Ltd. The purities of these standards were determined to be greater than 98% using HPLC coupled with a UV detector. Rheum palmatum L. (No. 190207), Magnolia officinalis Rehd. et Wils. (No. 190619), Citrus aurantium L. (No. 190128), and mirabilite (No. 190515) were purchased from Hubei Chenmei Herbal Pharmaceutical Co., Ltd. The samples were identified by Dr. Sen Li of Huazhong University of Science and Technology.
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4

Authentication of Herbal Medicines in MZRW

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All the six herbal medicines (HMR: RM130121-08; DH: 130524; BS: RM130128-11; KXR: RM121026-04; HP: RM121026-05; ZS: RM121210-11) contained in MZRW were supported and authenticated by PuraPharm Company, Nanning department (Nanning, China). The reference standards of gallic acid, amygdalin, paeoniflorin, hesperidin, neohesperidin, naringin, magnolol, honokiol, aloe emodin, physcion, chrysophanol were purchased from Shanghai Yuan-Ye Bio-technology company (Shanghai, China). Emodin, rhein, albiflorin were purchased from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). The purity of all these reference standards was ≥98.0% (HPLC). HPLC grade of acetonitrile, methanol and formic acid were purchased from Merck (Darmstadt, Germany). Deionized water was purified by the Millipore water purification system (Millipore, Milford, MA, United States). All other reagents used were of analytical grade. Twelve male Sprague-Dawley rats (220–250 g) were kindly provided by the Experimental Animal Center of Chinese University of Hong Kong (Hong Kong). The rats were bred in an environmentally controlled room (22 ± 2°C, relative humidity 50 ± 20%) with a natural light-dark cycle for 7 days before the experiment carried out. The animal study was carried out in accordance with the Guideline for Animal Experimentation of Hong Kong.
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5

Magnolol Preparation and Dosing

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Magnolol (CAS No: B21086) was obtained from Yuanye Biotech (Shanghai, China) with purity > 98%. Magnolol stock solution was prepared with dimethyl sulfoxide (DMSO) to 5 mM. The working solutions were diluted in E. Coli OP50 at final concentrations of 2.5, 5, 10, 20 and 50 μM.
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6

Magnolol Modulates Cell Cycle Proteins

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Human NSCLC cell lines PGCL3, SK-MES-1, NCI-H460, A549, NCI-1299 and immortalized human normal bronchial epithelium cell line 16HBE were purchased from ATCC [29] . A549 cells were cultured in Dulbecco's Modified Eagle Medium (Thermo Fisher Scientific). PGCL3, SK-MES-1, NCI-H460, NCI-1299 and 16HBE cells were maintained in RPMI-1640 medium (Thermo Fisher Scientific). Both mediums were supplemented with 10% fetal bovine serum (Wisent). All cells were incubated at 37°C with 5% humidified CO 2 . Mice were obtained from National Rodent Laboratory Animal Resources, Shanghai Branch of China. Cdc 2, p-Cdc 2, Cdc 25c, Cyclin B1 and actin antibodies were purchased from Cell Signaling Technology, and phospho-Ser/Thr-Pro MPM-2 antibody were obtained from Millipore. Magnolol with a purity of up to 98% was purchased from Shanghai Yuanye Bio-Technology Co., Ltd. Magnolol was dissolved in DMSO (Sigma) at a stock solution of 100 mM and stored at -20°C.
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