Lead acetate

The following antibodies were used in this study: anti-EZH2 (Cell Signaling, Beverly, MA, USA), anti-H3K27me3 (Millipore, MA, USA), anti-EED, anti-Vav1 (Proteintech, Wuhan, China). Acrylamide, bis-Acrylamide, β-mercaptoethanol, Hybond nitrocellulose, sodium dodecyl sulfate, Tris, and lead acetate (with the final concentration fixed to 1, 2.5, 5, 10, 20 μM, respectively) were purchased from Sinopharm Group (Beijing, China). All other reagents were of the highest analytical grade.

Chloroauric acid (99.99%), sodium hydroxide (99.7%), L-ascorbic acid (AA, 99.7%), sodium borohydride (96.0%), lead acetate (99.5%), thioacetamide (TAA, 99.0%), hexamethylenetetramine (HMT, 99.0%), hexadecyltrimethylammonium bromide (CTAB, 99.0%), and hydrochloric acid (36–38%) were purchased from Sinopharm Chemical Reagent Co. Ltd. (Shanghai, China). Deionized water with a resistivity of about 18.25 MΩ·cm was used as the solvent in all experiments.

Heavy metal compounds, lead acetate ((CH₃COO)₂Pb·3H₂O, AR ≥ 99.5%), cadmium dichloride (CdCl₂·2.5H₂O, AR ≥ 99.0%), nickel dichloride (NiCl₂·6H₂O, AR ≥ 98.0%), manganese dichloride (MnCl₂·4H₂O, AR ≥ 99.0%), zinc sulfate heptahydrate (ZnSO₄·7H₂O, AR ≥ 99.5%), cupric sulfate (CuSO₄·5H₂O, AR ≥ 99.0%), and potassium dichromate (K₂Cr₂O₇, AR ≥ 99.8%) were purchased from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China). Methyl mercury chloride was purchased from Ehrenstorfer GmbH (affiliation, Germany). The biochemical and hematological analysis were performed by Hitachi 7600-110 autoanalyzer (Hitachi Ltd, Tokyo, Japan) and blood analyzer (Sysmex XT-1800i; Sysmex Co, Kobe, Japan), respectively. Water maze testing was performed using a MWM video analysis system (RD1101-MWM-G, Shanghai Mobiledatum Information Technology Co., Ltd., China). The monoclonal antibodies including p38, P-p38, JNK, p-JNK (phosphorylated JNK), NFκB, Nrf2, Akt, and GAPDH were obtained from Cell Signaling Technology (Danfoss, MA, USA). Sprague Dawley (SD) rats were obtained from Zhejiang Provincial Laboratory Animal Science Center (Hangzhou, Zhejiang, China).

Leadacetate and chloroplatinic acid were purchased from Sinopharm Chemical Reagent company (Beijing, China). Graphene oxide nanocomposites solution (2 mg/mL) was purchased from Xianfeng Corporation (Nanjing, China). Nafion solution (20%), ascorbic acid (AA), uric acid (UA), dopamine (DA), 5-hydroxytryptamine (5-HT), dihydroxyphenylacetic acid (Dopac) and glutamate (Glu) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Saline (0.9% NaCl) was purchased from the ShuangHe Corporation (Beijing, China).

Standard chemicals, namely, danofloxacin, erythromycin, sulfadimidine, benzylpenicillin potassium, chloramphenicol, lincomycin, tetracycline, spectinomycin, thiamphenicol, dexamethasone, kanamycin, melamine, trimethoprim, zearanol, aflatoxin M₁, flunixin meglumine, and florfenicol, were purchased from Sigma (St. Louis, MO, USA). All the chemicals used were of analytical grade. Lead acetate, potassium oxalate, disodium hydrogen phosphate, trichloroacetic acid, sodium hydroxide, and dichloromethane were obtained from Sinopharm Chemical Reagent Co., Ltd. (China). High-purity water was obtained from a Milli-Q water system (Millipore, Billerica, MA, USA). Pure milk was purchased from a local supermarket. Commercialized semiquantitative ICA strip and an ICA strip reader were supplied by Beijing Meizheng Bio-Tech Co., Ltd. (Beijing, China)

Acetonitrile and formic acid (HPLC grade) were purchased from Merck (Darmstadt, GER). Analytic grade methanol was purchased from Fisher Scientific (Hudson, NH, USA). Deionized water was purified using a Milli-Q system (Millipore, MA, USA).
Standard compounds of cucurbitacin B (NO. 18052116), cucurbitacin D (NO. 19042168), and cucurbitacin E (NO. 19032512) were purchased from Shanghai Shifeng Bio-Technology Co., Ltd (Shanghai, China). Lead acetate (NO. 20160914), hydrochloric acid (NO. 20171107), soluble starch (NO. 20170414), iodine (NO. 20160914), and potassium iodide (NO. 20170110) were obtained from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China). p-Hydroxybenzyl hydrogen sulfite was prepared in our lab and confirmed by HR-MS and NMR analyses (Kang et al., 2017 (link)).
Fresh TR sample was collected from Shexian (Henan, China), the traditional “Dao Di” producing area, in November, 2018. 30 batches of commercial TR samples were collected from Bozhou material medicine market. All collected samples were identified as Trichosanthes kirilowii or Trichosanthes rosthornii by Prof. Lan-Ping Guo. The authenticated specimens were deposited in the National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences.