Hgcl2

Wistar rats were randomly divided into the following 4 groups: control, Lut (purity > 98%, Xi’an Weiao, Shanxi, China), HgCl₂ (Beijing Chemical Factory, China) and HgCl₂ + Lut groups, 7 rats each group. The rats received 80 mg/L HgCl₂ in their drinking water for 8 weeks. Freshly made HgCl₂-containing drinking water was provided every day. Luteolin was gavaged to rats once daily for the last 2 weeks at dose levels of 80 mg/kg in 1% DMSO, and the daily application volumes of luteolin were calculated based on the most recently recorded body weight of the individual animal. Twenty-four hours after the last feeding, surgery was performed on all rats under anesthesia to obtain the blood and liver tissue samples. For histological and TUNEL analysis, a portion of tissues was immersed in 4% paraformaldehyde overnight. The rest of samples were immediately frozen in liquid nitrogen and stored at −80°C until use.

The constructed plasmid pET21a-CaCDC42 containing the cdc42 gene of Candida albicans, which served as the template DNA, was recombined and prepared in the College of Life Science, Beijing Normal University, China. SYBR® Premix Ex Taq™ II containing Taq DNA polymerase catalyzing the elongation step of DNA replication, dNTPs, ROX Reference Dye II, DNA loading buffer, and SYBR Green II was purchased from Takara Bio Inc. (Dalian, China). The synthetic primers targeting cdc42 were provided by Sangon Biotech Co. Ltd. (Beijing, China) as follows: 5′-GTGATGGTGCCGTTGGTA-3′ (forward) and 5′-CCCTGTTCCTGGGTGATT-3′ (reverse), which produced a piece of DNA containing 397 bp. Thirteen heavy metal salts, ZnCl₂, Pb(NO₃)₂, CdCl₂·2.5H₂O, Ni(NO₃)₂·6H₂O, HgCl₂, CrCl₃, MnCl₂, CuSO₄, CoCl₂, Ce(NO₃)₃, Sr(NO₃)₂, Bi(NO₃)₃ and K₃Fe(CN)₆ were obtained from Beijing Chemical Reagent Company (Beijing, China). Ethylene diamine tetraacetic acid (EDTA), Ethidium bromide (EB), and agarose was from Sigma-Aldrich (St. Louis, USA). The DNA replication reactions in vitro were fulfilled and monitored using the Real-time PCR System ABI7500 (Applied Biosystems™, USA).

Citric acid was purchased from Tiantian Chemical Reagent Factory (Tianjin, China). p-phenylenediamine was obtained from Aladdin Reagent Co., Ltd. (Shanghai, China). Phenylalanine, arginine, glycine, threonine, aspartate, tryptophan, glutamate, and tyrosine were purchased from Tianjin Institute of Fine Chemical Industry Co., Ltd. (Tianjin, China). KCl, MgCl₂, CaCl₂, NaCl, MnSO₄, SrCl₂, CdSO₄, NiSO₄, and Fe₂(SO₄)₃ were obtained from Tianjin Kaitong Chemical Reagent Co., Ltd. (Tianjin, China). CuSO₄, HgCl₂, PbSO₄, ZnCl₂, AlCl₃, HCl, NaOH, and anhydrous ethanol were acquired from Beijing Chemical Plant (Beijing, China). Quinine sulfate was acquired from Beijing Enoch Technology Co., Ltd. (Beijing, China). All chemicals were analytical pure, and the experimental water was ultrapure water. Fetal bovine serum was obtained from Zhejiang Tianhang Biotechnology Co., Ltd. (Zhejiang, China).