Millipore purification system

Iodoacetamide (IAA) was purchased from Sigma-Aldrich Chemicals (St. Louis, Missouri, USA). XiaoErFuPi (XEFP) granules were obtained from Hunan Time Sun Pharmaceutical Co., Ltd (Yongzhou, China). Domperidone was obtained from Xi’an Janssen Pharmaceutical Ltd (Xi’an, China). All of the other chemicals were analytical grade reagents. The deionized water (R > 18.2 MΩ) used for all of the experiments was purified by using a Millipore purification system (Billerica, MA, USA).

Folin-Ciocalteu phenol reagent (FC), 2, 2-diphenyl-1-picrylhydrazyl (DPPH), porcine pancreatic α-amylase (EC 3.2.1.1), Saccharomyces cerevisiae α-glucosidase (EC 3.2.1.20), and porcine pancreas lipase (EC 3.1.1.3) were purchased from Sigma-Aldrich (St. Louis, MO, USA). All other chemicals were of analytical reagent grade. Authentic phenolic standards including caffeic, p-coumaric, ferulic and gallic acids, quercetin and (+)-catechin were obtained from Sigma-Aldrich (St. Louis, MO, USA). HPLC-grade methanol, acetonitrile and other solvents and reagents were purchased from Merck & Co. Inc. (NJ. USA) Deionized water prepared by Millipore purification system (Millipore Corp. Marlborough, MA, USA) was used for preparation of solutions.

Three genes encoding HFn, HFn‐PAS, and HFn‐PAS‐RGDK, respectively, were synthesized by the BGI Company (China). HFn‐PAS was constructed by fusing GFLG ASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPA GGSGG to HFn subunit C‐terminus through a flexible 15‐residue linker (GGGSGGGTGGGSGGG). Forty underlined residues constitute PAS peptide and GFLG is a Cathepsin B cleavable site. HFn‐PAS‐RGDK has four extra C‐terminus residues, RGDK, compared with HFn‐PAS. Three types of genes were inserted between NdeІ and XhoІ site of pET30a plasmid, and plasmids were then transformed into Escherichia coli (E. coli) BL21 (DE3) using heat‐shock transformation method.
Tryptone, yeast extract, Isopropyl β‐d‐thiogalactoside (IPTG) and kanamycin were purchased from Thermo Scientific (USA). Chromatography columns were bought from GE healthcare (USA). Doxorubicin hydrochloride (DOX) was purchased from Dalian Meilun Biotechnology (China). MCF7 cell line was kindly offered by Dr Nicholas Eyre from the University of Adelaide. RPMI‐1640 medium, penicillin‐streptomycin solution (100×), fetal bovine serum (FBS), and 0.25% trypsin‐EDTA (1×) solution were purchased from Thermo scientific (USA). All other reagents of analytical grade were purchased from Chem‐supply (Australia). Milli Q water was obtained from a Millipore purification system (Merck, USA) and used in all buffers and solutions.

Cyclohexanone analytical standards (purity ≥99.9%) were purchased from Dr. Ehrenstorfer (LGC Standards, Augsburg, Germany). Commercial triadimefon 20% emulsifiable concentrate was sourced from Jiangsu Sword Agrochemicals Co., Ltd. (Yancheng, China); tebuconazole 43% suspension was acquired from Qingdao Haina Biotechnology Co., Ltd. (Qingdao, China); and paclobutrazol 15% wet-table powder was obtained from Jiangsu Kesheng Group Co., Ltd. (Yancheng, China). Sodium chloride (NaCl) was provided by Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Saccharomyces cerevisiae powder was purchased from Yantai Di Boshi brewing machine Co., Ltd. (Yantai, China). The fermentation tanks were bought from Hebei Chaoya glass products Co., Ltd. (Hebei, China). Ultra-pure water was produced using a Millipore purification system (Millipore, Bedford, MA, USA). The grapes were obtained from Changxinghongyuan grape professional cooperative of Liaoning province and did not contain the target pesticides.

Chromatographic-grade acetonitrile, acetic acid, and methanol were purchased from Merck (Darmstadt, Germany). The water used as Milli-Q water was purified using a Millipore purification system (Millipore Corporation, Burlington， MA, USA). In this study, the standard lidocaine was bought from Shanghai New Asiatic Pharmaceuticals Co., Ltd. (Tianjin, China). All standards used in tests were stored in a −20 °C refrigerator in darkness.

To prepare Pt/ceria sample, 0.5 g of cerium(IV) oxide nanopowder (<25 nm) was dispersed in a solution consisting of 2.0 g of Urea and 8 mL of water. While stirring, 3.3 g of a solution (approximately 1 wt% Pt) of chloroplatinic acid hydrate in water was added to the dispersion. All chemicals were purchased from Sigma-Aldrich. Ultrapure water (18.2 MOhm) was provided by a Millipore purification system. The mixture was sealed in a vial and stirred for 24 h in an oil bath at 95 °C. Afterward, the postdeposition sample was separated and washed by 3 cycles of centrifuging/redispersing in water to remove the residual ions from the precursors. The sample was then dried overnight at 60 °C before being crushed into a powder and calcined at 500 °C (10 °C/min heating ramp). The resulting platinum loading of the sample was measured by Inductively Coupled Plasma (ICP) elemental analysis and determined to be 1.85 wt%. Note, certain commercial equipment, instruments, or materials are identified in this paper in order to specify the experimental procedure adequately. Such identification is not intended to imply recommendation or endorsement by the National Institute of Standards and Technology, nor is it intended to imply that the materials or equipment identified are necessarily the best available for the purpose.