Vancomycin

Mouse feed (D12450J) was purchased from Research Diets Inc. (New Brunswick, NJ, United States), and the diet composition was shown in Supplementary Table 1. Antibiotics including vancomycin, ampicillin, neomycin, and metronidazole were purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China).

Polymyxin B and ficin enzyme were obtained from Sigma-Aldrich (St. Louis, MO, USA). Polymyxin E was obtained from Kangmanlin (Nanjing, China). The following antibiotics were purchased from Yuanye Bio-technology Co., Ltd (Shanghai, China): erythromycin, lincomycin, vancomycin, ampicillin, ofloxacin, or Meilun (Dalian, China): linezolid, nisin, penicillin G, ciprofloxacin, tigecycline, and chlortetracycline.

The minimal inhibitory concentrations (MIC) for penicillin (β-lactams), cephalexin (β-lactams), ampicillin (β-lactams), ceftiofur (β-lactams), cefquinome (β-lactams), lincomycin (lincosamide class), oxytetracycline (Tetracycline class), marbofloxacin (Quinolone class), rifaximin (Rifamycin class), and vancomycin (Glycopeptides class) (Shanghai Yuanye Biotechnology Co., Ltd., Shanghai, China) were determined against 49 L. garvieae isolates using micro-broth dilution assays, following the Clinical Laboratory and Standards Institute guidelines [42 ]. All antimicrobial agents were used in concentrations ranging from 0.03 to 16μg/mL. S. aureus ATCC 29,213 was used as a quality control strain. Antimicrobial resistance was defined by combining intermediate and resistant categories into a single category. MDR was defined as resistance to ≥3 classes of antimicrobial agents [2 (link)]. Each experiment was performed in triplicate.

In spontaneous models, sex-matched WT and Kras mice aged 5 weeks were infected intranasally (i.n.) with HBLV-Cre-3xflag-ZsGreen-PURO Lenti-Cre (Hanbio, Shanghai, China) for induction of lung cancer as previously described [22 (link)]. The mice were fed with normal water or combined antibiotics, including 1 g/L ampicillin sodium (Sangon Biotech, Shanghai, China), 0.5 g/L vancomycin (Yuanye Bio-Technology, Shanghai, China), 1 g/L neomycin trisulfate salt hydrate (Sangon Biotech, Shanghai, China), and 1 g/L metronidazole (Sangon Biotech, Shanghai, China) in their drinking water 5 weeks before Lenti-Cre delivery, which was continued throughout the whole experiment. All experimental procedures were approved by the Ethics Committee of the University of Science and Technology of China (Hefei, China).

The antimicrobial susceptibility test of the isolated C. difficile was performed using the agar dilution method recommended by the US Clinical and Laboratory Standards Institute (CLSI) M11-A8 (36 ). The following eight antibiotics were included in this study: vancomycin, clindamycin, erythromycin, levofloxacin, moxifloxacin, ciprofloxacin, meropenem, and metronidazole (Yuan ye Biotech., Shanghai, China). The results were interpreted with reference to the susceptibility breakpoints for anti-anaerobic drugs in CLSI 2017 version M100-S27 (37 ). Furthermore, for antibiotics with no susceptibility breakpoints in the CLSI document, the susceptibility breakpoints were proposed according to Huang's report (16 (link)).

Hinokitiol (>99.0% GC content), formate (≥98% HPLC), and chlorTetracycline were purchased from Aladdin Bio-Chem Technology Co., Ltd. (Shanghai, China). Vancomycin, ciprofloxacin, erythromycin, and minocycline were purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (China). Tetracycline and tigecycline were purchased from Shanghai Macklin Biochemical Co., Ltd. (China). Acetonitrile (HPLC) was purchased from Tedia Co., Inc. (USA).
S. aureus ATCC 43300, S. aureus ATCC 29213, S. aureus ATCC 25923, E. coli ATCC 35218, and B. subtilis 168 were preserved in our laboratory. Clinical isolates, including S. aureus ZY-1, S. aureus ZY-2, S. aureus ZY-3, S. aureus ZY-4, S. aureus ZY-5, and S. aureus ZY-6 were obtained from patients at the First Affiliated Hospital of Zhejiang University, Hangzhou, China. All bacterial strains were grown routinely on MH medium (Oxoid, UK).

Mannuronate oligosaccharides (MOS) were purchased from Qingdao BZ Oligo Biotech Co., Ltd. (Qingdao, China). Potassium oxonate (PO), allopurinol, 0.5% Sodium carboxymethyl cellulose (CMC-Na) solution, vancomycin, ampicillin, metronidazole, neomycin, 2-ethylbutyric acid, acetic acid (Ace), propionic acid (Pro), butyric acid (But), isobutyric acid (Isobut), valeric acid (Val) and isovaleric acid (Isoval) were purchased from Shanghai yuanye Bio-Technology Co., Ltd. (Shanghai, China). Animal feeds were purchased from Trophic Animal Feed High-Tech Co., Ltd. (Nantong, China). Serum uric acid (UA), blood urea nitrogen (BUN), adenosine deaminase (ADA), and xanthine oxidase (XOD) assay kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Elisa kits for interleukin-1β (IL-1β), interleukin-12 (IL-12) and interleukin-18 (IL-18) were purchased from Suzhou Calvin Co., Ltd. (Suzhou, China). The primers XOD, ADA, GLUT9, and URAT1 were purchased from Sangon Biotech (Shanghai) Co., Ltd. (Shanghai, China). Protein antibodies URAT1, GLUT9, and ABCG2 were purchased from ABclonal Technology Co., Ltd. (Wuhan, China). All other chemicals and reagents used were of analytical quality.