Journal: Frontiers in Immunology

Article Title: Neuroinflammatory and transcriptional dynamics during SARS-CoV-2 infection in KRT18-hACE2 mouse brain

doi: 10.3389/fimmu.2026.1716597

Figure Lengend Snippet: Neuropathological changes and inflammatory mediators in K18-hACE mouse brain following SARS-CoV-2 infection. (A, B) Immunofluorescence staining of brain tissue from uninfected and SARS-CoV-2 infected mice showing colocalization of DAPI (blue, nuclear stain) with (A) Iba-1 (green) and (B) CD68 (red). Lower panels show single staining for Iba-1 or CD68. Scale bar: 50 µm. (C) TUNEL assay of brain tissue from uninfected and SARS-CoV-2 infected mice. (D) Quantification of panel C showing the percentage of TUNEL+ cells in each group. Student t-test was performed to determine the significance. **p < 0.01, ***p < 0.001).

Article Snippet: DNA fragmentation was detected on rehydrated paraffin-embedded sections using a one-step TUNEL in situ apoptosis detection kit (E-CK-A325; Elabscience), according to the manufacturer’s instructions.

Techniques: Infection, Immunofluorescence, Staining, TUNEL Assay

Journal: Journal of Nanobiotechnology

Article Title: Targeting delivery of miR-146a via IMTP modified milk exosomes exerted cardioprotective effects by inhibiting NF-κB signaling pathway after myocardial ischemia-reperfusion injury

doi: 10.1186/s12951-024-02631-0

Figure Lengend Snippet: MEs-miR-146a protects H9c2 cells from OGD/R induced damage. ( A ) After 24 h of incubation, PKH26 labeled exosomes could be uptaken up by H9c2. Scale bar = 20 μm. ( B ) MEs-miR-146a decreased OGD/R induced cell apoptosis of H9c2 according to TUNEL assay. Scale bar = 50 μm. * P < 0.05, ** P < 0.01 versus the control group; & P < 0.05, && P < 0.01 versus the OGD/R group; ▲ P < 0.05 versus the OGD/R-MEs group. n = 6

Article Snippet: The one-step TUNEL Apoptosis Detection Kit (C1088, Green Fluorescence, Beyotime) was used to incubate for 60 min at RT, following by nuclear labeling with DAPI (H-1200-10, Vector Laboratories, Burlingame, CA, USA), both in the dark.

Techniques: Incubation, Labeling, TUNEL Assay, Control

Journal: Journal of Nanobiotechnology

Article Title: Targeting delivery of miR-146a via IMTP modified milk exosomes exerted cardioprotective effects by inhibiting NF-κB signaling pathway after myocardial ischemia-reperfusion injury

doi: 10.1186/s12951-024-02631-0

Figure Lengend Snippet: MEs-miR-146a protects NRCM cells from OGD/R induced damage. ( A ) Representative fluorescence images indicated that PKH26 labeled exosomes were taken up by NRCM cells after 24 h of incubation. Scale bar = 20 μm. ( B ) MEs-miR-146a decreased OGD/R induced cell apoptosis of NRCM according to TUNEL assay. Scale bar = 50 μm. ** P < 0.01 versus the control group; & P < 0.05, && P < 0.01 versus the OGD/R group; ▲ P < 0.05 versus the OGD/R-MEs group. n = 6

Article Snippet: The one-step TUNEL Apoptosis Detection Kit (C1088, Green Fluorescence, Beyotime) was used to incubate for 60 min at RT, following by nuclear labeling with DAPI (H-1200-10, Vector Laboratories, Burlingame, CA, USA), both in the dark.

Techniques: Fluorescence, Labeling, Incubation, TUNEL Assay, Control

Journal: Journal of Nanobiotechnology

Article Title: Targeting delivery of miR-146a via IMTP modified milk exosomes exerted cardioprotective effects by inhibiting NF-κB signaling pathway after myocardial ischemia-reperfusion injury

doi: 10.1186/s12951-024-02631-0

Figure Lengend Snippet: MEs-miR-146a exhibited better therapeutic efficacy in decreasing apoptotic cells and limiting inflammation than MEs. ( A - B ) Representative TUNEL staining images and quantification of apoptotic radio. ** P < 0.01 versus the Sham group; & P < 0.05 and && P < 0.01 versus the MIRI group. ( C - F ) The expression of inflammatory factors of rat serum were detected by ELISA. * P < 0.05, ** P < 0.01 versus the Sham group; & P < 0.05, && P < 0.01 versus the MIRI group; ▲ P < 0.05, ▲▲ P < 0.01 versus the MIRI-MEs-miR-146a group. n = 5

Article Snippet: The one-step TUNEL Apoptosis Detection Kit (C1088, Green Fluorescence, Beyotime) was used to incubate for 60 min at RT, following by nuclear labeling with DAPI (H-1200-10, Vector Laboratories, Burlingame, CA, USA), both in the dark.

Techniques: Drug discovery, TUNEL Assay, Staining, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Journal of Nanobiotechnology

Article Title: Targeting delivery of miR-146a via IMTP modified milk exosomes exerted cardioprotective effects by inhibiting NF-κB signaling pathway after myocardial ischemia-reperfusion injury

doi: 10.1186/s12951-024-02631-0

Figure Lengend Snippet: Tail vein injection of IMTP-MEs-miR-146a significantly ameliorated myocardium apoptosis and attenuated inflammation at the early stage of MIRI. ( A ) Representative H&E images of heart tissue from each group. Scale bar = 50 μm. ( B - C ) Representative images of TUNEL staining 24 h after MIRI and quantitative analysis of apoptotic radio. * P < 0.05 and ** P < 0.01 versus the MIRI group; & P < 0.05 versus the MIRI-MEs-miR-146a group. n = 8. ( D - G ) The expression of inflammatory factors of rat serum were detected by ELISA. ∆∆ P < 0.01 versus the MIRI group; ▲▲ P < 0.01 versus the MIRI-MEs-miR-146a group. n = 8

Article Snippet: The one-step TUNEL Apoptosis Detection Kit (C1088, Green Fluorescence, Beyotime) was used to incubate for 60 min at RT, following by nuclear labeling with DAPI (H-1200-10, Vector Laboratories, Burlingame, CA, USA), both in the dark.

Techniques: Injection, TUNEL Assay, Staining, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Chinese Medicine

Article Title: Honokiol acts as an AMPK complex agonist therapeutic in non-alcoholic fatty liver disease and metabolic syndrome

doi: 10.1186/s13020-023-00729-5

Figure Lengend Snippet: AMPK activation is independent of classical pathways. A Western blots of p-AMPKα, AMPKα, p-ACC, ACC, p-LKB1, LKB1, p-CAMKK2, CAMKK2, p-TAK1, TAK1, and PP2C of primary hepatocytes subjected to PA stimulation at the indicated conditions. n = 3 replicates. B Western blots of p-AMPKα, AMPKα, p-ACC, ACC, p-LKB1, LKB1, p-CAMKK2, CAMKK2, p-TAK1, TAK1, and PP2C in the livers of HFD-fed mice treated with vehicle or honokiol. n = 5 mice per group. C Western blots of p-AMPKα, AMPKα, p-ACC, ACC, p-LKB1, LKB1, p-CAMKK2, CAMKK2, p-TAK1, TAK1, and PP2C in the livers of CDAHFD-fed mice treated with vehicle or honokiol. n = 5 mice per group. D Western blots of p-AMPKα, AMPKα, p-ACC, ACC, p-LKB1, LKB1, p-CAMKK2, CAMKK2, p-TAK1, TAK1, and PP2C in the livers of MCD-fed mice treated with vehicle or honokiol. n = 5 mice per group. E Representative image of the oxygen consumption rate (OCR) of primary hepatocytes subjected to DMSO or honokiol at the indicated conditions. n = 3 replicates. Omy, oligomycin, F1F0 ATP synthase inhibitor. FCCP, mitochondrial uncoupler. Rot, retenone, complex I inhibitor. AA, antimycin A, complex II inhibitor. F ATP, ADP, and AMP in the livers of HFD-fed mice treated with vehicle or honokiol. n = 6 mice per group. Student’s t -test was applied for statistical analysis. G ATP, ADP, and AMP in the livers of CDAHFD-fed mice treated with vehicle or honokiol. n = 6 mice per group. Student’s t -test was applied for statistical analysis. H ATP, ADP, and AMP in the livers of MCD-fed mice treated with vehicle or honokiol. n = 6 mice per group. Student’s t -test was applied for statistical analysis

Article Snippet: Primary antibodies targeting PP2C (#ab211660) were procured from Abcam (Cambridge, UK).

Techniques: Activation Assay, Western Blot

Journal: Cancer Biology & Medicine

Article Title: The inducible secreting TLR5 agonist, CBLB502, enhances the anti-tumor activity of CAR133-NK92 cells in colorectal cancer

doi: 10.20892/j.issn.2095-3941.2023.0033

Figure Lengend Snippet: CD133 has upregulated expression in colorectal cancer tissues. (A) The HColA-180Su11 microarray (Outdo Biotech, Shanghai, China) containing 76 primary colon cancer samples was stained with an anti-CD133 antibody to determine the expression of CD133. Left: paracarcinoma tissue with negative CD133 expression; middle: primary colon cancer samples with lower CD133 expression; right: primary colon cancer samples with higher CD133 expression. Representative images (magnification ×4) are shown for CD133 immunostaining. The scale bar represents 500 μm. (B) The percentage of different levels of CD133 expression is indicated. (C) CD133 expression on colon cancer cell lines was analyzed by flow cytometry. (D) The HOrgN090PT02 microarray (Outdo Biotech) containing 25 human normal tissue samples was immunostained with an anti-CD133 antibody to determine the expression of CD133; human normal tissues are negative for CD133. Representative staining image fields (magnification ×20) are shown. Scale bars represent 100 μm.

Article Snippet: To evaluate the surface expression of CD133 in colon cancer cell lines (SW620, SW480, and HCT116), the cells were washed with 1X PBS containing 4% BSA and incubated with PE-conjugated anti-CD133 antibody (ab253271; Abcam); IgG isotype controls were included.

Techniques: Expressing, Microarray, Staining, Immunostaining, Flow Cytometry

Journal: Cancer Biology & Medicine

Article Title: The inducible secreting TLR5 agonist, CBLB502, enhances the anti-tumor activity of CAR133-NK92 cells in colorectal cancer

doi: 10.20892/j.issn.2095-3941.2023.0033

Figure Lengend Snippet: CAR133-NK92 cells with inducible secretion of CBLB502 enable superior proliferation, activation, and antitumor activity. (A) The absolute number of CAR133-NK92 and CAR133-i502-NK92 cells was analyzed when the cells were stimulated with CD133+ and CD133− antigens for different lengths of time (days). Data represent the mean ± SD of quadruplicate wells, * P < 0.05; NS, not significant. (B) The activation marker, CD69, was quantified by flow cytometry when CAR133-NK92 or CAR133-i502-NK92 cells were cultured with SW620 and SW480 cells for 24 h. The mean values from triplicate cultures are shown. *** P < 0.001. (C) Effector cells (NK92, CAR133-NK92, and CAR133-i502-NK92 cells) were co-cultured with colorectal cancer cells at an E:T ratio of 10:1 for 24 h. Effector cells were then harvested and CD107a expression was analyzed by flow cytometry. The mean values from triplicate cultures are shown. * P < 0.05; ** P < 0.01; NS, not significant. (D) Cell killing by CAR133-NK92 and CAR133-i502-NK92 cells was investigated by the LDH cytotoxicity assay after co-culture with target cells at the indicated E:T ratios. All data are expressed as the means ± SEMs of triplicate samples. * P < 0.05; NS, not significant. (E) The cytotoxicity of CAR133-NK92 and CAR133-i502-NK92 cells against colon cancer cells was calculated by RTCA. (F) CAR133-NK92 and CAR133-i502-NK92 cells were co-cultured with SW620 cells for 5 days, and the cytokines and chemokines in the culture supernatant were determined by the human cytokine array C5 (Ray Biotech, Atlanta, GA, USA). * P < 0.05. (G) The concentrations of GM-CSF and IL-7 in the supernatant were further determined by specific ELISA kits. Data represent the mean+SEM. * P < 0.05; ** P < 0.01.

Article Snippet: To evaluate the surface expression of CD133 in colon cancer cell lines (SW620, SW480, and HCT116), the cells were washed with 1X PBS containing 4% BSA and incubated with PE-conjugated anti-CD133 antibody (ab253271; Abcam); IgG isotype controls were included.

Techniques: Activation Assay, Activity Assay, Marker, Flow Cytometry, Cell Culture, Expressing, LDH Cytotoxicity Assay, Co-Culture Assay, Enzyme-linked Immunosorbent Assay

Journal: Cancer Biology & Medicine

Article Title: The inducible secreting TLR5 agonist, CBLB502, enhances the anti-tumor activity of CAR133-NK92 cells in colorectal cancer

doi: 10.20892/j.issn.2095-3941.2023.0033

Figure Lengend Snippet: CBLB502 promotes the antitumor activity of CAR133-NK92 cells in mice with subcutaneous CD133-expressing colon tumors. (A) Schedule of the challenge protocol. Mice were inoculated with CD133+ SW620 cells (1 × 10 6 cells/mouse) followed 14 days later by CAR133-i502-NK92, CAR133 NK92, or NK92 cells (5 × 10 6 cells/mouse, n = 5/group). (B) Colon cancer growth was monitored by in vivo bioluminescence imaging. (C) Kaplan–Meier survival curves showing the tumor-free survival of tumor-bearing mice treated with CAR133-i502-NK92, CAR133-NK92, or NK92 cells. Significance was determined by the log-rank test. *** P < 0.001. (D) Blood collected on day 15 was analyzed by flow cytometry for the presence of effector cells, CAR133-i502-NK92 cells showed enhanced proliferation in vivo . *** P < 0.001. (E) The number of CD56+ cells, representing NK92 cells, was increased in tumor samples of mice treated with CAR133-i502-NK92 cells compared to CAR133-NK92 and NK92 cells on day 15 after effector cell treatment. Each scale bar represents 50 μm. (F) Absolute number of CD56+ cells in . Data represent the mean ± SD of quadruplicate wells. *** P < 0.001.

Article Snippet: To evaluate the surface expression of CD133 in colon cancer cell lines (SW620, SW480, and HCT116), the cells were washed with 1X PBS containing 4% BSA and incubated with PE-conjugated anti-CD133 antibody (ab253271; Abcam); IgG isotype controls were included.

Techniques: Activity Assay, Expressing, In Vivo, Imaging, Flow Cytometry

Journal: Cancer Biology & Medicine

Article Title: The inducible secreting TLR5 agonist, CBLB502, enhances the anti-tumor activity of CAR133-NK92 cells in colorectal cancer

doi: 10.20892/j.issn.2095-3941.2023.0033

Figure Lengend Snippet: On-target off-tumor toxicities in the mouse model treated with CAR133-i502-NK92 cells. (A) The expression of mouse CD133 protein was tested in mouse stomach tissue using IHC assays. Representative staining image fields (magnification ×200) are shown. Scale bars represent 100 μm. (B) Histopathologic analysis of mouse organ tissues by H&E staining. Magnification ×20. Each scale bar represents 500 μm. (C) Detection of CAR-NK92 cells infiltrated into stomach tissues on day 15 after effector cell treatment by IHC staining of CD56 showed that there was minimal immune cell infiltration into the stomach (magnification ×100), The scale bar represents 200 μm. (D) Body weight of the individual mice from each treatment group. Data are presented as the mean ± SD of three independent experiments. Error bars represent the standard deviation.

Article Snippet: To evaluate the surface expression of CD133 in colon cancer cell lines (SW620, SW480, and HCT116), the cells were washed with 1X PBS containing 4% BSA and incubated with PE-conjugated anti-CD133 antibody (ab253271; Abcam); IgG isotype controls were included.

Techniques: Expressing, Staining, Immunohistochemistry, Standard Deviation

Journal: Cancer Biology & Medicine

Article Title: The inducible secreting TLR5 agonist, CBLB502, enhances the anti-tumor activity of CAR133-NK92 cells in colorectal cancer

doi: 10.20892/j.issn.2095-3941.2023.0033

Figure Lengend Snippet: Adoptively transferred CAR133-i502-NK92 cells successfully control tumor development in mice with CD133+ and CD133− mixed colon tumor models. (A) Schedule of the in vivo experimental design using CD133+ and CD133- mixed colon tumor model. (B) BLI illustrating colon cancer growth. (C) Kaplan–Meier survival curves generated from the survival of mice treated with CAR133-i502-NK92 and CAR133-NK92 cells. *** P < 0.001. (D) Expression of CD31 protein was tested in the mouse tumor tissue using IHC assays. Representative photomicrographs are shown (magnification ×400). The scale bar is 50 μm. *** P < 0.001. (E) Representative immunostaining images of CD3+ T-cell infiltration into cancer tissues. Representative staining of tumor sections in each experimental group is shown (magnification ×400). The scale bar represents 50 μm. *** P < 0.001.

Article Snippet: To evaluate the surface expression of CD133 in colon cancer cell lines (SW620, SW480, and HCT116), the cells were washed with 1X PBS containing 4% BSA and incubated with PE-conjugated anti-CD133 antibody (ab253271; Abcam); IgG isotype controls were included.

Techniques: Control, In Vivo, Generated, Expressing, Immunostaining, Staining

Journal: Cancer Biology & Medicine

Article Title: The inducible secreting TLR5 agonist, CBLB502, enhances the anti-tumor activity of CAR133-NK92 cells in colorectal cancer

doi: 10.20892/j.issn.2095-3941.2023.0033

Figure Lengend Snippet: The schematic diagram of the anti-tumor activity of CAR133-NK92 cells in colorectal cancer. We engineered CAR133-NK92 cells with inducible secretion of CBLB502 and demonstrated that CAR133-i502-NK92 cells not only specifically eliminate CD133-positive colon cancer cells in a CAR133-dependent manner, but also indirectly eradicate CD133-negative colon cancer cells in a CBLB502-specific endogenous immune response manner. The figure was created with Biorender.com .

Article Snippet: To evaluate the surface expression of CD133 in colon cancer cell lines (SW620, SW480, and HCT116), the cells were washed with 1X PBS containing 4% BSA and incubated with PE-conjugated anti-CD133 antibody (ab253271; Abcam); IgG isotype controls were included.

Techniques: Activity Assay