Ham s f 12k medium

MoS₂ crystal was purchased from Muke Nano Science and Technology Co., Ltd. (Nanjing, China). HS-PEG-NH₂ (molecular weight (MW): 5000) was provided by Xi'an Ruixi biological Co., Ltd. (Xi'an, China). 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT), Hoechst 33,258, and Ham’s F12K medium were obtained from Sigma-Aldrich (St. Louis, USA). Trypsin-ethylenediaminetetraacetic acid (EDTA) and fetal bovine serum (FBS) were purchased from Gibco-BRL (Burlington, Canada). An Annexin V-APC/7-AAD apoptosis detection kit was supplied by Biolegend Company (San Diego, USA). All other reagents were purchased from J&K Scientific, Ltd., and used without further purification.

The human lung cancer alveolar A549 cells and normal lung fibroblast WI 38VA13 Subline 2RA cells were obtained from Bioresource Collection and Research Centre (BCRC, Taiwan). Cells were cultured in Ham’s F-12K medium (Sigma-Aldrich, St. Louis, MO, USA) with 10% fetal bovine serum (FBS, Gibco) at 37°C in a humidified atmosphere containing 5% CO₂. For the experiments, the cells were treated with SBPE for several time points or treated with SBPE for 6 h followed by the additional chemotherapy drugs exposure for additional time points.

A total of 20 gastric cancer tissues and their corresponding adjacent normal tissues were collected from 20 gastric patients who underwent curative resections. These cancer tissue specimens and adjacent normal tissue specimens were routinely confirmed by biopsy and stored in liquid nitrogen. The study group consisted of 14 male and 6 female patients, with a median age of 60.12 years. No primary tumors from other sites were observed for these gastric patients. Prior to surgery, the gastric patients received no other treatment. The human gastric cancer AGS cell line was purchased from the cell center of Shanghai Life Science Institutes of the Chinese Academy of Sciences (Shanghai, China) and was cultured with 10% fetal bovine serum (Hyclone, Shanghai, China) and Ham’s F12K medium Sigma-Aldrich (St. Louis, MO, USA) at 37°C and 5% CO₂. This study was performed at the First Hospital of Zhangjiagang (Zhangjiagang, China) and also approved by the Institutional Review Board of the First Hospital of Zhangjiagang. Written informed consent was obtained from each patient.

Human gastric adenocarcinoma cell line AGS (CRL-1739; American Type Culture Collection, Manassas, VA) was cultured in Ham’s F-12K medium (Sigma-Aldrich, St Louis, MO) supplemented with 10% fetal calf serum (Gibco-BRL, Rockville, MD) and 2mM glutamine at 37°C under 5% CO2. Where indicated, glutamine-free F-12K medium was also used. Primary gastric epithelial cells were isolated from human gastric biopsies and cultured as described previously [9 (link),26 (link)]. All cells were co-cultured with 3 day-old H. pylori WT or the different isogenic mutants (Δggt, ΔvacA, ΔureAB, ΔvacA/ggt, ΔureAB/ggt) at a multiplicity of infection (MOI) of 1:100 [9 (link)]. Where indicated, cells were treated with rGGT at an approximately equivalent amount to that produced by the viable bacteria [9 (link)].

A549 human pulmonary epithelial cell line was purchased from the American Type Culture Collection (Manassas, VA, USA). Enoxaparin was obtained from Aventis Pharma Ltd. (NSW, Australia). UFH was purchased from Hospira Pty. Ltd. (Victoria, Australia). Sodium chloride, tetrahydrofuran, sodium hydroxide, N-methyl-N-(trimethylsilyl) trifluroacetamide, methanol, acetic acid, Ham’s F12K medium, antibiotics (penicillin G and streptomycin), trypsin-ethylenediaminetetraacetic acid (EDTA), trypsin, thrombin, enzyme-linked immunosorbent assay (ELISA) kits for IL-6 and IL-8, trypan blue exclusion assay kit, lactate dehydrogenase (LDH) activity assay kit, and potassium phosphate monobasic and dibasic were purchased from Sigma-Aldrich (Castle Hill, NSW, Australia). Fetal bovine serum, IL-6 and IL-8 recombinant human proteins were obtained from Invitrogen (Grand Island, NY, USA). Deuterium oxide (D₂O) was purchased from Cambridge Isotope Laboratories (Andover, MA, USA).