Human BMSCs were purchased from Cyagen (USA) and cultured in human BMSCs basal media (Cyagen, USA) with 10% fetal bovine serum (Cyagen, USA), 1% penicillin-streptomycin (Cyagen, USA), and glutamine (Cyagen, CA, USA). And osteogenic induction was conducted by BMSCs osteogenic differentiation media (Cyagen, USA) with 10% fetal bovine serum (Cyagen, USA), glutamine (Cyagen, CA, USA), 1% penicillin-streptomycin (Cyagen, USA), ascorbic acid (Cyagen, CA, USA), β-Glycerolphosphate disodium (Cyagen, USA), and dexamethasone (Cyagen, USA). All the cells were cultured at 37°C in saturated humidity containing 5% CO2. The media were changed every two days. The BMSCs were digested using 0.25% trypsin (Solarbio, China), when they grew to a confluence of 70–80%. To investigate the effect of topical antibiotics, BMSCs were cultured without antibiotics (Con group) or with antibiotics, which further subdivided into three subgroups: Van group: cultured with vancomycin (Solarbio, China), (1g per person11 (link),12 (link) = 14.28μg/mL), Tob group: cultured with tobramycin (Solarbio, China), (2g per person3 (link),13 (link)=28.57μg/mL), Van+Tob group (vancomycin 14.28μg/mL combined with tobramycin 28.57μg/mL).
Vancomycin
Manufactured by Solarbio
Sourced in China
Vancomycin is a laboratory product used for the detection and identification of vancomycin-resistant microorganisms in clinical samples. It is a glycopeptide antibiotic that inhibits bacterial cell wall synthesis.
Automatically generated - may contain errors
Lab products found in correlation
Metronidazole, by Solarbio (10 mentions)
Ampicillin, by Solarbio (8 mentions)
Neomycin sulfate, by Solarbio (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Neomycin, by Solarbio (4 mentions)
Streptomycin, by Solarbio (2 mentions)
Amphotericin b, by Solarbio (2 mentions)
Ciprofloxacin, by Solarbio (2 mentions)
Bhi broth, by Hopebio (2 mentions)
Dmem f12, by Thermo Fisher Scientific (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Sodium butyrate, by Solarbio (1 mentions)
Pam3csk4 p3c, by InvivoGen (1 mentions)
Lb broth, by Thermo Fisher Scientific (1 mentions)
Isopropyl β d 1 thiogalactopyranoside (iptg), by Sangon (1 mentions)
Bhi broth, by Thermo Fisher Scientific (1 mentions)
Bicinchoninic acid assay kit, by Beyotime (1 mentions)
Mrs broth, by Solarbio (1 mentions)
Yeast extract, by Solarbio (1 mentions)
Tryptone, by Solarbio (1 mentions)
22 protocols using vancomycin
1
Effects of Topical Antibiotics on BMSC Osteogenesis
2
TNBS-Induced Colitis Model Protocol
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TNBS (P2297–10 ml) was obtained from Sigma-Aldrich (MO, USA). VDZ (28713–300 mg) was purchased from Takeda Pharmaceuticals (Tokyo, Japan). Vancomycin, metronidazole, ampicillin, and neomycin sulfate were obtained from Solarbio Technology (Beijing, China; purity＞95%). All BA standards were obtained from Shanghai ZZBIO (Shanghai, China; purity＞95%). Fetal bovine serum, DMEM/F12, and TRIzol reagent were purchased from Thermo Fisher Scientific (MA, USA). All reagents for protein measurement and extraction were purchased from Beyotime Biotechnology (Shanghai, China).
3
Antibiotic-induced microbiome depletion model
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Commensal microbes were depleted using antibiotics as previously reported32 (link). Specifically, four kinds of antibiotics including ampicillin (1 g/l) (Solarbio), vancomycin (0.5 g/l) (Solarbio), neomycin sulfate (1 g/l) (Solarbio) and metronidazole (1 g/l) (Solarbio) were dissolved in sterile water and stored in 4 °C no more than a week before using. This antibiotic-contained water was supplied as drinking water to pregnant, breastfeeding mice and was changed every 3 days. Meanwhile, the offspring mice from pregnant C57BL/6 dams without antibiotic treatment were maintained in parallel as controls in each experiment. For early-Abx model, 8-week-old control or early-Abx male mice (18~22 g body weight) were subjected to HCC models, 16 S rRNA sequencing and microbiota metabolic or liver NK cell analysis (5~11 mice per group, born from 2~4 different dams and fed in 2~3 different cages).
4
Immune Modulation via Pharmacological Agents
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Ampicillin, neomycin sulfate, metronidazole, vancomycin, vitamin A, and sodium butyrate were purchased from Solarbio life science (Beijing, China). TLR2 ligands Pam3CSK4 (P3C) was purchased from InvivoGen (Hongkong, China). Recombinant chicken GM-CSF protein (rGM-CSF) and recombinant chicken IL-17 protein (rIL-17) were purchased from Abcam (Cambridge, MA, UK).
5
Recombinant Protein Expression Workflow
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PCR primers with sequences were synthesized by Genomics institution (China). FastPfu DNA polymerase, Escherichia coli strains DH5α and BL21 (DE3) were purchased from TransGen Biotech (China). Quickcut restriction endonucleases NotI and NdeI were both purchased from Takara Bio (Japan). T4 DNA ligase was purchased from BioLab (U.S.A.). IPTG was purchased from Sangon (China). DNA marker, protein marker and live/dead bacteria staining kit were all purchased from Thermo Fisher Scientific (U.S.A.). LB broth and BHI broth were purchased from Oxoid (U.K.). Bicinchoninic acid assay kit was purchased from Beyotime Biotechnology (China). Vancomycin was purchased from Solarbio Life Sciences (China).
6
Modulating Gut Microbiome to Enhance Viral Immunity
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To eliminate as much as possible the intestinal microbial background of mice, ampicillin (1 mg/ml), streptomycin (5 mg/ml), vancomycin (0.25 mg/ml), and mycomycin (1 mg/ml; Solarbio) were added to the sterile drinking water of mice for 6 days, followed by FMT. Mouse feces from the PBS + H9N2 and 0111 + H9N2 groups were collected on the seventh day and processed as described previously (Yang et al., 2021 (link)). Briefly, fecal samples were placed in a sterile N2 incubator, mixed thoroughly, and then plugged into a cell filter to collect bacteria and centrifuged at 12,000 rpm for 10 min. The samples were subsequently added to 10% glycerol, stored at −80°C. FMT before replacing the glycerol with PBS and stained with methylene blue staining to count the number of viable microorganisms. Mice were gavaged in 100 μl volumes (n = 5). One day later, all mice were intranasally attacked with H9N2 (106.5 EID50), and their survival and weight loss were monitored 14 days after infection. FMT experiment was repeated twice independently.
7
Microbiome Restoration via Probiotic Inoculation
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Mice were cohoused for 3–4 weeks before antibiotic treatment to allow for equilibration of the microbiota. Broad-spectrum antibiotic consisted of metronidazole (1 g/l), neomycin (1 g/l), ampicillin (1 g/l), vancomycin (0.5 g/l) and streptomycin (1 g/l) (all from Solarbio, Beijing, China) was administered in the drinking water for 1 week and then was replaced with regular water for the rest of the experiment.
Mice were orally inoculated 24 h later with 1 × 109 CFUs Lactobacillus salivarius. L. salivarius was grown at 37°C in MRS broth (Solarbio) and were washed twice with PBS prior to use.
Mice were orally inoculated 24 h later with 1 × 109 CFUs Lactobacillus salivarius. L. salivarius was grown at 37°C in MRS broth (Solarbio) and were washed twice with PBS prior to use.
8
Biopolymer-Based Microcapsule Synthesis
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Petroleum ether (PE), sodium
carbonate (Na2CO3), anhydrous ethanol, sodium
chloride (NaCl), and isopropyl alcohol were obtained from Sinopharm.
Lithium bromide (LiBr) and Span-80 were purchased from Aladdin. Branched
polyethylenimine (PEI, Mw ∼ 25,000) was acquired from Sigma-Aldrich.
Polyethylene glycol 8000 (PEG-8000), vancomycin, yeast extract, tryptone,
agar power, DNase I, phosphate-buffered saline (PBS), penicillin/streptomycin,
and a CCK-8 assay kit (CA1210) were purchased from Solarbio. A live/dead
staining kit (L34951) was obtained from Thermo Fisher Scientific.
FITC and rhodamine B were purchased from Macklin.
carbonate (Na2CO3), anhydrous ethanol, sodium
chloride (NaCl), and isopropyl alcohol were obtained from Sinopharm.
Lithium bromide (LiBr) and Span-80 were purchased from Aladdin. Branched
polyethylenimine (PEI, Mw ∼ 25,000) was acquired from Sigma-Aldrich.
Polyethylene glycol 8000 (PEG-8000), vancomycin, yeast extract, tryptone,
agar power, DNase I, phosphate-buffered saline (PBS), penicillin/streptomycin,
and a CCK-8 assay kit (CA1210) were purchased from Solarbio. A live/dead
staining kit (L34951) was obtained from Thermo Fisher Scientific.
FITC and rhodamine B were purchased from Macklin.
9
Investigating MRSA Virulence through AtlA
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MRSA strain USA300 (WT) and its mutant strain after it knocked out gene atlA (ΔatlA) were grown in Tryptic Soy Broth (TSB, Coolaber Science&Technology Co., Ltd. China) medium at 37°C. The reagents included ZJ-2 (1,3-bis(4-(trifluoromethyl)phenyl)urea), vancomycin (Solarbio Science&Technology Co., Ltd. China), Miller-Hinton broth (MHB, Thermo Fisher Scientific Inc. USA), Tris-Cl, Triton X-100 (Labgic Science&Technology Co., Ltd. China), Bacteria PG ELISA kit (Chuzhou shinuoda Biological Technology Co., Ltd. China), MDP ELISA kit (Nanjing SenBeiJia Biological Technology Co., Ltd. China), Mouse IL-6 and TNF-α ELISA kit (Elabscience Biotechnology Co., Ltd. China).
10
Culturing and Identifying Helicobacter pylori
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Helicobacter pylori (H. pylori, Sydney strain or SS1) was provided by Professor Young-Joon Surh (Seoul National University, Seoul, South Korea). H. pylori SS1 was grown on Columbia Agar (Oxoid, Basingstoke, Hampshire, England, Cat number CM0331) plates for 48 h at 37 °C under microaerophilic conditions on 7% lysed horse blood (Solarbio, Beijing Solarbio Science & Technology, Beijing, China Cat number S9050) agar and antibiotics, including amphotericin B (1.5 μg/mL; Solarbio, Cat number: A8250), trimethoprim (1.25 μg/mL) (Tichea Chemical Industry Development Co, Shanghai, China, Product code T2286), and vancomycin (2.5 μg/mL) (Solarbio, Cat number: V8050). Identification was performed by Gram staining (Solarbio, Cat number G1060) and testing for urease (Nanjing Jiancheng Bioengineering Institute, Nanjing, China; Cat number C013-2), catalase (Nanjing Jiancheng Bioengineering Institute, Cat number A007-1), and molecular markers.
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