Berberine

GO nanosheets were purchased from Xianfeng Materials Technology (Nanjing, China). All the DNA oligonucleotides used in this study were synthesized and purified by Shanghai Sangon Biotechnology Co., Ltd. (Shanghai, China). The transpeptidase region (extracellular region) of PBP2a was provided by Shanghai Sangon Biotechnology Co., Ltd. via the Escherichia coli prokaryotic expression system. SYBR Green qRT-PCR Kit and SYTO9 dye were obtained from Baiaolaibo Biotechnology Co., Ltd. (Beijing, China). UNIQ-10 Column TRIzol Total RNA Isolation Kit, ELISA plate, 96-well tissue culture plate, lambda exonuclease, and berberine were purchased from Shanghai Sangon Biotechnology Co., Ltd. All chemicals used were of analytical grade. All reagents were dissolved in ultrapure water. The MRSA strain was provided by the Medical Laboratory Center of the First Affiliated Hospital of Hunan University of Chinese Medicine (Changsha, China). S. aureus, S. epidermidis, S. typhimurium, and E. coli strains were obtained from Xiangya Hospital of Central South University (Changsha, China).

Berberine was obtained from Sangon Biotech Co., Ltd. (Shanghai, China). Azoxymethane (AOM) was purchased from Sigma Aldrich (St. Louis, MO, USA). Dextran sulfate sodium salt (DSS) was provided by Dalian Meilun Biotech Co., Ltd. (Dalian, China). The AIN-93M rodent diet was purchased from Trophic Animal Feed High-tech Co., Ltd. (Nantong, China). All antibodies used in this study, were obtained from Zhengneng Biotechnology Co., Ltd. (Chengdu, China).

The M. canis strain (CMCC(F)M3h) used in this study was provided by the Institute of Internal Medicine, Chinese Academy of Medical Sciences (Nanjing, China) National Center. The M. canis was cultured in Tryptic Soy Broth (TSB) at 28°C for 96 h. The densities of the suspension were adjusted to primary inoculum of 1.0 × 10⁷ CFU/mL in the normal saline solution with total volume of 100 mL in conical flasks. berberine hydrochloride (BC) (HPLC>98%) was purchased from Yuan Ye Biological technology co., Ltd, Shanghai, China. The BC treated group (three samples) was subject to medicinal stress dose which means 1/2 MIC (minimum inhibitory concentration) dose (1 mg/mL berberine in DMSO), while the control group was fed with DMSO (Purity>99%) which was purchased from Sangon Biotech co., Ltd Shanghai, China only. Each group has three repeats. After 6 h incubation (28°C,150 rpm), the seeds were filtered with gauze and frozen after treatment for RNA isolation and further analysis.