Journal: Acta pharmaceutica Sinica. B

Article Title: A cyclodextrin-based nanoformulation achieves co-delivery of ginsenoside Rg3 and quercetin for chemo-immunotherapy in colorectal cancer.

doi: 10.1016/j.apsb.2021.06.005

Figure Lengend Snippet: Figure 2 Ginsenoside Rg3 induced immunogenic cell death in CRC cells. (A) IC50 of Rg3 for CT26 and HCT116 cells at 24 h. Data are presented as mean SD (n Z 3). (B) Apoptosis (%) in CT26 and HCT116 cells following treatment of Rg3 at 6, 12 and 24 h. Data are presented as mean SD (n Z 3). *P < 0.05 and **P < 0.01 relative to DMSO. (C) The activity of UPR signaling pathways following treatment of Rg3 ([c] Z 30 mmol/L) at 6, 12 and 24 h. The quantification was demonstrated in Fig. S1. (D) The characterization of ICD in CRC cells following treatment of Rg3 ([c] Z 30 mmol/L), including CRT exposure (6 h), ATP secretion (12 h) and HMGB1 release (12 h). Data are presented as mean SD (n Z 3). *P < 0.05 relative to DMSO; scale bar Z 20 mm. (E) The expression of CD11c and CD86 in DCs stimulated with the supernatant from Rg3 ([c] Z 30 mmol/L) -treated cells (24 h). Data are presented as mean SD (n Z 3). **P < 0.01 relative to DMSO. (F) The in vivo vaccination assay using BALB/C and nude mice. Data are presented as mean SD (n Z 4). ***P < 0.001.

Article Snippet: DNA fragments were detected using the TransDetect Fluorescein TUNEL Cell Apoptosis Kit (TransGen Biotech), and nuclei were stained with DAPI (Beyotime Biotech), for confocal microscopic analysis (Olympus); 2) Measurement of immune cells.

Techniques: Activity Assay, Protein-Protein interactions, Expressing, In Vivo

Journal: Acta pharmaceutica Sinica. B

Article Title: A cyclodextrin-based nanoformulation achieves co-delivery of ginsenoside Rg3 and quercetin for chemo-immunotherapy in colorectal cancer.

doi: 10.1016/j.apsb.2021.06.005

Figure Lengend Snippet: Figure 3 Quercetin caused reactive oxygen species in CRC cells. (A) IC50 of QTN for CT26 and HCT116 cells at 24 h. Data are presented as mean SD (n Z 3). (B) Apoptosis (%) in CT26 and HCT116 cells following treatment of QTN ([c] Z 80 mmol/L) at 24 h. Data are presented as mean SD (n Z 3). **P < 0.01 relative to DMSO. (C) The activity of Bcl-2/BAX/caspase 9/caspase 3 signaling pathways following treatment of QTN ([c] Z 80 mmol/L) at 6, 12 and 24 h. The quantification was demonstrated in Fig. S2. (D) The ROS level in CT26 and HCT116 cells following treatment of QTN ([c] Z 80 mmol/L) at 6, 12 and 24 h. Data are presented as mean SD (n Z 3). **P < 0.01 and ***P < 0.001 relative to DMSO. (E) Cell viability (%) of CT26 and HCT116 cells with or without NAC prior to treatment of QTN ([c] Z 80 mmol/L) (24 h). Data are presented as mean SD (n Z 3). *P < 0.05 and **P < 0.01 relative to untreated control. (F) Apoptosis (%) in CT26 and HCT116 cells with or without NAC prior to treatment of QTN ([c] Z 80 mmol/L) (24 h). Data are presented as mean SD (n Z 3). *P < 0.05 and **P < 0.01 relative to untreated control.

Article Snippet: DNA fragments were detected using the TransDetect Fluorescein TUNEL Cell Apoptosis Kit (TransGen Biotech), and nuclei were stained with DAPI (Beyotime Biotech), for confocal microscopic analysis (Olympus); 2) Measurement of immune cells.

Techniques: Activity Assay, Protein-Protein interactions, Control

Journal: Acta pharmaceutica Sinica. B

Article Title: A cyclodextrin-based nanoformulation achieves co-delivery of ginsenoside Rg3 and quercetin for chemo-immunotherapy in colorectal cancer.

doi: 10.1016/j.apsb.2021.06.005

Figure Lengend Snippet: Figure 4 Synergistic effects of Rg3 and QTN in CT26 cells. (A) IC50 of drug combination at 24 h. Data are presented as mean SD (n Z 3). CI values at IC50 were shown in Fig. S3. (B) Apoptosis (%) caused by drug combination at 24 h. Data are presented as mean SD (n Z 3). *P < 0.05 and **P < 0.01 relative to DMSO. (C) The CRT exposure with or without NAC before treatment of drug combination (6 h). Data are presented as mean SD (n Z 3). *P < 0.05 and **P < 0.01 relative to DMSO; scale bar Z 20 mm. (D) The ATP secretion with or without NAC before treatment of drug combination at 12 h. Data are presented as mean SD (n Z 3).*P < 0.05 and **P < 0.01, between NAC and No NAC. (E) The HMGB1 release with or without NAC before treatment of drug combination at 12 h. Data are presented as mean SD (n Z 3). *P < 0.05 and **P < 0.01, between NAC and No NAC. (F) The expression of CD11c and CD86 in DCs stimulated (24 h) by the supernatant from Rg3- treated cells with or without pretreatment of NAC. Data are presented as mean SD (n Z 3). *P < 0.05, **P < 0.01 and ***P < 0.001, relative to DMSO.

Article Snippet: DNA fragments were detected using the TransDetect Fluorescein TUNEL Cell Apoptosis Kit (TransGen Biotech), and nuclei were stained with DAPI (Beyotime Biotech), for confocal microscopic analysis (Olympus); 2) Measurement of immune cells.

Techniques: Expressing

Journal: Acta pharmaceutica Sinica. B

Article Title: A cyclodextrin-based nanoformulation achieves co-delivery of ginsenoside Rg3 and quercetin for chemo-immunotherapy in colorectal cancer.

doi: 10.1016/j.apsb.2021.06.005

Figure Lengend Snippet: Figure 8 Combination therapy of targeted co-formulation and Anti-PD-L1 for CRC. (A) Treatment schedule and IVIS images. (B) The CRC progression over a 35-day period. Data are presented as mean SD (n Z 5). *P < 0.05 and **P < 0.01; NS, no significance. (C) Animal survival (median survival: PBS ~38 days, Anti-PD-L1 ~40 days, targeted co-formulation ~62 days, and combination z 96 days). Data are presented as mean SD (n Z 5). **P < 0.01 and ***P < 0.001. (D) Immunofluorescent staining assay (green Z DNA fragments and blue Z nuclei) on Day 20 to assess apoptosis in the tumor (scale bar Z 50 mm). Data are presented as mean SD (n Z 3). *P < 0.05 and **P < 0.01, relative to PBS. (E) Level of immune cells in the tumor on Day 20 was analyzed using flow cytometry (BD). Data are presented as mean SD (n Z 4). *P < 0.05 and **P < 0.01; NS, no significance. (F) The mRNA expression of cytokines and chemokines in the tumor on Day 20 was analyzed using real time RT-PCR. Data are presented as mean SD (n Z 4). *P < 0.05 and **P < 0.01; NS, no significance. (G) Orthotopic CRC mice treated with targeted co-formulation following the removal of CD4 þ or CD8þ T cells. Data are presented as mean SD (n Z 4). *P < 0.05 and **P < 0.01; NS, no significance.

Article Snippet: DNA fragments were detected using the TransDetect Fluorescein TUNEL Cell Apoptosis Kit (TransGen Biotech), and nuclei were stained with DAPI (Beyotime Biotech), for confocal microscopic analysis (Olympus); 2) Measurement of immune cells.

Techniques: Formulation, Staining, Cytometry, Expressing, Quantitative RT-PCR