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8 protocols using trimethylamine

1

Characterization of Seed Nutrients in Chinese Pumpkin Cultivars

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Seven commercial cultivars of seed‐rich pumpkin in China, including Dali, Luoren No.2, Luoren No.3, Jinhui No. 4, Ruifeng No.9, Yinhui No. 3, and Xuecheng No.2 (coded LR‐1, LR‐2, LR‐3, JH‐4, RF‐9, YH‐3, and XC‐2), were purchased from Mudanjiang Beidahuang Food Co., Ltd. All seeds were received in good shape (without damage) and were stored at 4℃.
Gamma‐aminobutyric acid (GABA) and phenylisothiocyanate (PITC) were purchased from Sigma Chemical Co. Monosodium glutamate (MSG), trimethylamine, n‐hexane, methyl alcohol, acetonitrile of HPLC grade, and other analytical reagents were obtained from Sinopharm Chemical Reagent Co., Ltd.
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2

Synthesis and Characterization of Composite Nanofibers

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Silver nitrate (AgNO3, 99.0%), anhydrous ethylene glycol (EG, 99.8%), ammonium persulphate (APS, 98.0%), concentrated sulfuric acid (H2SO4, 98.0%), magnesium oxide (MgO, 99.9%), boric acid (99.8%), methyl red, methylene blue, trimethylamine (TMA·H2O, 33.0%), ammonium hydroxide (NH3·H2O, 25–28%), ethanol (C2H6O, 99.8%) and sodium carbonate (Na2CO3, 99.8%) were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China), Polyvinyl pyrrolidone (PVP, K29-32) and aniline (C6H7N, 99.5%) were purchased from Aladdin Industrial, and aniline was purified by vacuum distillation. Silkworm cocoons were provided by Shanghai Natural Wild-insect Kingdom Co. Ltd. (Shanghai, China). Dry air was purchased from Jiangsu Thorpe Group Co. Ltd. (Zhenjiang, China). The morphology characteristic of composite nanofibers were observed by scanning electron microscope (SEM, JSM-7800F, JEOL, Tokyo, Japan). Raman spectra were collected by XploRA Plus, HORIBA. Ltd. (Longjumeau, France).
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3

Synthesis of Polyurethane Emulsion Polymers

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Acylic acid (AA), methacrylic acid (MAA), butyl acrylate (BA), n-butyl methacrylate (BMA), styrene (St), hydroxyethyl methacrylate (HEMA), 1-dodecanethiol, ammonium persulfate (APS), trimethylamine (TEA), ammoniumhydroxide (NH3∙H2O), sodium dodecyl sulfonate (SDS), dodecanethiol, N,N-dimethylethanolamine (DMEA), 1,4-Butylene glycol (BDO), butanedioic acid and Alkylphenolpolyoxyethylene ether (OP-10) were provided by Sinopharm Chemical Reagent Co., Ltd. (SCRC, Shanghai, China). Reactive emulsifier SE-10 was purchased from Foshan Kodi Gas Chemical Industry Co., Ltd (Foshan, China). Isophoronediisocyanate (IPDI), polycarbonate diol (PCDL, the type of polycarbonate diol was 1,5-pentanediol and 1,6-hexanediol) were provided by Zhongshan Daoqum Co., Ltd. (Zhongshan, China). Dimethylolpropionic acid (DMPA) was purchased from Shanghai Aladdin Co., Ltd. (Shanghai, China). Dibutyltindilaurate (DBTDL) was purchased from Beijing Sananhua Co., Ltd. (Beijing, China). In order to eliminate the inhibitor, the activated carbon adsorption method and alkali washing with NaOH or NaHCO3 were used for the water-soluble monomer (AA, MAA, HEMA) and other monomer (BA, BMA, St), respectively. Other materials were used without further purification.
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4

Spinal Cord Clearing via FDISCO Protocol

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The FDISCO protocol was performed according to the original literature (Qi et al., 2019 (link)). Before clearing, tetrahydrofuran (THF, Sigma-Aldrich, St. Louis, MO, United States) and dibenzyl ether (DBE, Sigma-Aldrich, St. Louis, MO, United States) were preprocessed by column absorption chromatography with basic activated aluminum oxide (Sinopharm Chemical Reagent Co., Ltd., Shanghai, China) to remove the peroxides. The spinal cords were dehydrated in THF solutions diluted in dH2O (pH adjusted to 9.0−9.5 with trimethylamine; Sinopharm Chemical Reagent Co., Ltd., Shanghai, China) at graded concentrations as follows: 50, 70, 80, and 100 vol% (twice) for 45−60 min at each step. After dehydration, pure DBE was used as a refractive index matching solution to clear the spinal cord. During the clearing process, the spinal cord was placed in a glass chamber under the dark. All steps were performed at 4°C with slight shaking.
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5

Formulation Development of ZMT Tablets

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ZMT was obtained from Energy Chemical Co., Ltd. (Shanghai, China). PU and MDX were purchased from Freda Biotechnology Co., Ltd. (Linshu County, Linyi, China) and COFCO biochemical energy Co., Ltd. (Longjiang County, Qigihaer, China), respectively. Propylene glycol (PG) and polyethylene glycol 400 (PEG) were obtained from Yonghua Chemical Technology Co., Ltd. (Zhitang District, Changshu, China) and Ling Feng Chemical Reagent (Shanghai, China), respectively. Trimethylamine, glycerin (GLY), anhydrous sodium hydrogen phosphate, and potassium dihydrogen phosphate were received from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). HPLC grade acetonitrile and methanol were bought from Fisher Scientific Worldwide (Shanghai, China). ZMT® tablets were purchased from Xuhui Pharmaceutical Co., Ltd. (Chengdu, China).
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6

Bromination of Poly(2,6-dimethyl-1,4-phenylene oxide)

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Poly(2,6-dimethyl-1,4-phenylene
oxide) (PPO) was purchased from Sigma-Aldrich Co., Ltd., P. R. China.
2,2′-Azobisisobutyronitrile (AIBN) and N-bromosuccinimide
(NBS) were purchased from Adamas Reagent Co., Ltd., China. Chlorobenzene
was bought from Shanghai Titas Co., Ltd., P. R. China. Potassium chromate
(K2CrO4), trimethylamine, N-methylimidazole, sodium sulfate (Na2SO4), N-methypyrrolidine, sodium hydroxide (NaOH), N-methylpiperidine, chloroform (CHCl3), sodium chloride
(NaCl), silver nitrate (AgNO3), ethanol (EtOH), and 1-methyl-2-pyrrolidone
(NMP) were obtained from Sinopharm Chemical Reagent Co., Ltd., P.
R. China and were utilized as attained. Deionized (DI) water was utilized
throughout the work.
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7

Antioxidant and Cytotoxic Evaluation of Linyi Leaves

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The leaves of LLJ were collected in Linyi, Shandong Province, and identified by Prof. Jixiang He (College of Pharmacy, Shandong University of Traditional Chinese Medicine). A voucher specimen (20180701006) has been deposited at the Shandong Analysis and Test Centre. All chemicals, including trifluoroacetic acid, trimethylamine, 2,2-Diphenyl-1-pyridine hydrazide (DPPH), butylated hydroxytoluene (BHT), silver nitrate for AgNPs synthesis, and anhydrous were analytical grade and purchased from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China). All aqueous solutions were prepared using deionized water produced by the Milli-Q osmosis system (Millipore, USA). HeLa cell lines were bought from the American Type Culture Collection. HepG2 cell lines were purchased from the Shanghai Gefan Biotechnology Co., Ltd (GF589). MDA-MB-231 cells were obtained from the Nanjing Cobioer Biosciences Co., Ltd (CBP60382).
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8

Cytotoxicity Assays in Cell Lines

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DMF was distilled from CaH2. All other commercial solvents and reagents were used without further purification unless otherwise mentioned. 3/4-Nitrophthalonitrile, n-butylamine, 1,3-diphenylisobenzofuran (DPBF), Li, and extra dry n-butanol were purchased from Energy Chemical Co., Ltd. Trimethylamine was purchased from Sinopharm Chemical Reagent Co., Ltd. Human serum albumin (HSA) and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) were purchased from Sigma-Aldrich (Shanghai) Trading Co. Ltd. Phosphate-Buffered Saline (PBS) and Dulbecco's Phosphate-Buffered Saline (DPBS) were purchased from Biological Industries USA, Inc. RPMI Medium 1640 basic (1X) (RPMI 1640), Fetal Bovine Serum (FBS), Penicillin–Streptomycin mixtures (Pen–Strep) and trypsin–EDTA solution were purchased from HyClone Laboratories, Inc. 2′,7′-Dichlorodihydrofluorescein Diacetate (DCFH-DA), Calcein Acetoxymethyl Ester (Calcein-AM) and propidium iodide (PI) were purchased from Shanghai Mackinlin Biochemical Co., Ltd. All organic solvents were purchased from Sinopharm Chemical Reagent Co., Ltd. Deionized water prepared with an Aquapro System (18 MΩ) was employed in all experiments.
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