XJR was purchased from Anhui Bozhou Traditional Chinese and Western Medicine Co., Ltd. (Anhui, China). XJR is composed of 7 different herbal ingredients: Hedyotis diffusa (20 g), Radix pseudostellariae (15 g), Bombyx batryticatus (10 g), Cremastra appendiculata (10 g), Centipede (3 g), Radix ophiopogonis (12 g), and Akebia trifoliata Koidz (12 g). Decoction of XJR (2 g/mL) was prepared according to the standard method,8 (link) diluted with saline (v/v, 1:1), and stored at 4°C for later use. Cisplatin was purchased from the Shanghai Yuanye Biotechnology Co., Ltd (Shanghai, China). MTT, dimethyl sulfoxide (DMSO), PBS, FBS, penicillin/streptomycin, and the other reagents for cell culture were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Luciferase reporter plasmids were purchased from Genechem Co., Ltd (Shanghai, China). Antibodies for Notch1 (b8925) and β-actin (20,536-1-AP) were purchased from Abcam (Cambridge, UK). BCA Protein Assay Kits were purchased from ThermoFisher Scientific (Waltham, MA).
Cisplatin
Manufactured by Yuanye Bio-Technology
Sourced in China
Cisplatin is a laboratory equipment product used for various scientific applications. It is a platinum-based compound that has a core function of inhibiting cell division and inducing apoptosis in cells. The detailed specifications and intended use of this product are not provided within the scope of this request.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Thermo Fisher Scientific (1 mentions)
Luciferase reporter plasmids, by Genechem (1 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Oleanolic acid, by Merck Group (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Nadph, by Merck Group (1 mentions)
2 chloroadenosine, by Merck Group (1 mentions)
Nitric oxide assay kit, by Nanjing Jiancheng (1 mentions)
Gsh assay kit, by Beyotime (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Lyso tracker red, by Beyotime (1 mentions)
Hoechst 33342, by Beyotime (1 mentions)
Calcein am pi staining kit, by Beyotime (1 mentions)
17β estradiol e2, by Cayman Chemical (1 mentions)
Copper acetate, by Maclin Biochemical Technology (1 mentions)
Annexin 5 fitc pi, by Beyotime (1 mentions)
H2o2 assay kit, by Solarbio (1 mentions)
9 protocols using cisplatin
1
Anticancer Herbal Decoction XJR
2
Honokiol Modulates Cellular Metabolism
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Cisplatin (Platinum content > 98%) was purchased from Shanghai Yuanye Biotechnology Co. (Shanghai, China); MgIG Injections were provided by Chia Tai Tianqing Pharmaceutical Group Co. (Lianyungang, Jiangsu, China); MgIG standard was obtained from National Institutes for Food and Drug Control (Beijing, China); oleanolic acid, NAD+, NADH, NADPH, NR, NMN, NAM, 2-chloroadenosine were bought from Sigma-Aldrich (St Louis, MO, USA). Penicillin-streptomycin, DMEM-F12, and trypsin were acquired from Gibco (Grand Island, NY, USA); Fetal bovine serum (FBS) was procured from Genetimes Biotechnology Co. (Shanghai, China); PCR reaction primers were purchased from Invitrogen (Shanghai, China). Honokiol (HKL) was purchased from Selleck (Shanghai, China). The remaining reagents were commercially available.
3
Fabrication of Cisplatin-Loaded 3D Titanium Implants
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The hydrogel was prepared as previously described [44 (link),45 (link)]. PLGA (1500–2000)-PEG (1000–1500)-PLGA (1500–2000) thermosensitive hydrogel (Shanghai Yuanye Bio-Technology Co., Ltd., Shanghai, China) was added to deionised water while stirring; the polymer to water mass ratio was 1:4. After incubation for 7 nights at 4 °C, the gel completely dissolved and formed a clear viscous solution (Fig. S1D in Supporting Information). The gel was then loaded with a specific amount of cisplatin (Shanghai Yuanye Bio-Technology Co., Ltd.) (Fig. S1E in Supporting Information). The 3D-printed titanium alloy implants were placed inside a gel chamber, and equal volumes of cisplatin/hydrogel were injected into the pores at 4 °C (Fig. S1B in Supporting Information). The samples were warmed to room temperature (18–25 °C) before implantation to form the cisplatin/hydrogel-loaded 3D-printed Ti6Al4V implants (Figs. S1C and F in Supporting Information). The materials and components of the different complexes are listed in Table 1 .Table 1
cisplatin/hydrogel-loaded 3D-printed Ti6Al4V implants: the materials and their components.
| Material | Component (w/w) |
|---|---|
| cisplatin | Pt (purity: 65%) |
| PLGA-PEG-PLGA hydrogel | PLGA-PEG-PLGA (20.0%), Deionised water (80.0%) |
| 3D-printed titanium alloy implants | Ti (90.0%), Al (5.7%), V (3.8%) |
4
Copper-based Nanoparticle Anticancer Protocol
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Copper acetate was obtained from Macklin. Gallic acid (GA) was purchased from Kermel. Surfactant Aerosol OT was obtained from Alfa Aesar. Cisplatin was purchased from Shanghaiyuanye Bio-Technology Co., Ltd. l -arginine (l -Arg), NADH, glutathione (GSH) and H2O2 Assay Kit were obtained from Beijing Solarbio Science & Technology Co., Ltd. DSPE-PEG2000 was purchased from Top-Peptide Co., Ltd. BCA Kit, Lyso-Tracker Red, Hoechst 33342, DAPI, GSH Assay Kit, Calcein-AM/PI staining kit and Annexin V-FITC/PI were obtained from Beyotime Biotechnology. Charcoal-stripped fetal bovine serum (CS-FBS) was supplied by Hyclony. Nitric Oxide Assay Kit was obtained from Nanjing Jiancheng Bioengineering Institute. NO fluorescence probe (DAF-FM DA) was purchased from meilunbio. 4-hydroxyestradiol (4-OHE2) and 17β-estradiol (E2) were obtained from Cayman Chemical. PD98059 and Calpeptin were purchased from MCE. All other chemicals were provided by Sigma Aldrich (St. Louis, MO, USA) unless mentioned otherwise.
5
Cell Growth Assay with Dox and Cisplatin
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The in vitro growth of cells treated with Dox (Yuanye Biotech, Shanghai, China) or Cisplatin (DDP, Yuanye Biotech, Shanghai, China) was detected with a CCK8 assay kit (Sangon Biotech, Shanghai, China) according to the manufacturer’s instructions. The cell growth curve was generated from the corresponding normalized OD450 values.
6
Phytochemical Analysis and Anticancer Activity
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The fresh PPL was collected from Hangzhou City (Zhejiang, China) in 01 September, 2020. These samples were identified by Professor Wang Ping from Zhejiang University of Technology. The voucher specimens (No. PPL 20200923) were deposited in Moganshan Campus of Zhejiang University of Technology. The leaves were air−dried naturally at room temperature, crushed, and passed through an 80−mesh sieve. After that, they were stored in a −18 °C freezer until use.
The standards of polyphyllin I (≥98%), II (≥98%), VI (≥98%), and VII (≥95%); dioscin (≥98%); kaempferol 3-O-gentiobioside (≥95%); isorhamnetin-3-O-glucoside (≥98%); and cisplatin were purchased from Shanghai Yuanye Biotechnology Co. Ltd., (Shanghai, China). Ultrapure water was produced by a Barnstead TII Pure Water System (Waltham, MA, USA). All other analytical−grade chemicals used in this experiment were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). A549 cells, MCF−7 cells, HepG2 cells, A431 cells, and HBE cells were purchased from Hunan Fenghui Biotechnology Co. Ltd. Roswell Park Memorial Institute (RPMI−1640). Dulbecco’s Modified Eagle Medium (DMEM) and fetal bovine serum (FBS) were obtained from Gbico (New York, NY, USA).
The standards of polyphyllin I (≥98%), II (≥98%), VI (≥98%), and VII (≥95%); dioscin (≥98%); kaempferol 3-O-gentiobioside (≥95%); isorhamnetin-3-O-glucoside (≥98%); and cisplatin were purchased from Shanghai Yuanye Biotechnology Co. Ltd., (Shanghai, China). Ultrapure water was produced by a Barnstead TII Pure Water System (Waltham, MA, USA). All other analytical−grade chemicals used in this experiment were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). A549 cells, MCF−7 cells, HepG2 cells, A431 cells, and HBE cells were purchased from Hunan Fenghui Biotechnology Co. Ltd. Roswell Park Memorial Institute (RPMI−1640). Dulbecco’s Modified Eagle Medium (DMEM) and fetal bovine serum (FBS) were obtained from Gbico (New York, NY, USA).
7
Cisplatin and EPI Combination Treatment
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Cisplatin and EPI were purchased from Shanghai Yuanye Bio-Technology Co. Ltd (Shanghai, China). MK2206, LY294002, GSK690693, KU-55933, RI-1, MG132, VE-821, aphidicolin, and 3-MA were purchased from Selleck Chemicals LLC (Shanghai, China). IRB2 and cycloheximide were purchased from MedChemExpress (Shanghai, China). Thymidine were purchased from Abcam (Cambridge, UK). Lipofectamine 2000 reagent, Opti-MEM, and IP lysis buffer were purchased from Invitrogen (Shanghai, China).
8
Nanomaterial-based Cisplatin Delivery
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Tannic acid (TA), Pluronic® F68, cisplatin (CDDP) and Rhodamine B (RB) were purchased from Yuanye biotechnology company (Shanghai, China). Silver nitrate was obtained from aladdin reagent Co., Ltd. (Shanghai, China). Sodium Dodecyl Sulfonate (SDS) was bought from Sigma-Aldrich (USA). Phosphorylated H2AX (γH2AX) antibody was supplied by Cell Signaling Technology company. NOX4, and NAPDH antibody were purchased from Proteintech company. H2O2 fluorescent probe BES H2O2-Ac was got from Wako (Cat. #021-17801 (1 mg), Japan). MQAE probe and Fe2+ probe (Ferro Orange) were bought from Beyotime company (Shanghai, China) and Dong Ren company (Shanghai, China), respectively.
9
Evaluating CTPG's Anticancer Potential
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The antitumor effects of CTPG on HepG2 and BEL-7404 cells were assessed using MTT (3-(4, 5-dimethyl-2thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide). HepG2 and BEL-7404 cells were plated into 96-well plates (5×10 4 cells/well) and treated with 0, 200, 400 and 600 μg/mL CTPG for 24 h and 48 h respectively after 24 h of incubation at 37℃. 35 μg/mL cisplatin (Yuanye) was used as positive control. 100 uL MTT (0.5 mg/mL, diluted by medium without FBS medium) was added to each well and cultured for 3 h at 37℃ and 5% CO 2 . After incubation, Loading [MathJax]/jax/output/CommonHTML/fonts/TeX/fontdata.js the plates were centrifuged at 1200 rpm for 7 min, the medium was removed and 200 μg/mL DMSO was added to each well to dissolve the formed formazan crystals. The OD490 values were measured by a 96-well microplate reader (Bio-Rad Laboratories, CA, USA). To evaluate the effects of CTPG on splenocytes, the cells were isolated from C57BL/6 mice and plated into 96-well plates at a density of 1×10 5 cells/well. Splenocytes were treated with 0, 200, 400 and 600 μg/mL for 24 h, 48 h and 72 h, respectively. The cell viability was calculated as the followed formula: Cell viability (%) = (OD treated /OD untreated ) × 100%.
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