Journal: International Journal of Nanomedicine

Article Title: Engineered Cancer Cell Membrane-Functionalized Metal-Organic Frameworks for Irinotecan/Curcumin Codelivery in Colorectal Cancer: Enhanced Efficacy and Mitigated Toxicity

doi: 10.2147/IJN.S558675

Figure Lengend Snippet: MZCC characterization. ( A ) UV−Vis absorbance spectra of ZIF-8, CPT-11, CCM, ZIC and ZICC. ( B ) FT-IR of CCM, CPT-11, ZIF-8, ZIC and ZICC. ( C ) XRD of CCM, ZIF-8, ZIC, ZICC and MZCC. ( D ) TEM image of ZIC, ZICC and MZCC. Scale bar: 100 nm. ( E ) Dynamic light scattering of ZIC, ZICC and MZCC. ( F ) Zeta potentials of ZIC, ZICC and MZCC. ( G ) SDS-PAGE of marker, MCM, ZICC and MZCC.

Article Snippet: Irinotecan was purchased from Aladdin Scientific Co., Ltd. (Shanghai, China).

Techniques: SDS Page, Marker

Journal: The Journal of clinical endocrinology and metabolism

Article Title: Circulating Exosomes Activate Dendritic Cells and Induce Unbalanced CD4+ T Cell Differentiation in Hashimoto Thyroiditis.

doi: 10.1210/jc.2019-00273

Figure Lengend Snippet: Figure 3. HT exosomes bind to and upregulate TLR2/TLR3. DCs were cocultured with HT exosomes or HC exosomes for 24 hours. (a) Immunofluorescence analysis showed that DCs (blue color, marked by DAPI) were colocalized with HT exosomes or HC exosomes (green color, marked by PKH-67) after coculture for 24 hours. TLR2 and TLR3 were marked by Alexa Fluor 594 (red color) (magnification 3400, confocal laser-scanning microscopy). PBMCs were cocultured with HT exosomes or HC exosomes for 24 hours. (b) Representative patterns of CD11c1TLR21, CD11c1TLR31, and CD11c1TLR41 cells as analyzed by flow cytometry. (c) Frequency of CD11c1TLR21, CD11c1TLR31, and CD11c1TLR41 cells in the HT exosomes and HC exosomes groups (n 5 20 per group). The PBS group was used as a negative control (n 5 4). Statistical analyses of multiple groups were performed by Kruskal-Wallis H-test or one-way ANOVA. *P , 0.05; **P , 0.01.

Article Snippet: To perform the inhibition test, according to the literature and the results of our preliminary experiments, DCs were preincubated with the TLR2 antagonist PAb hTLR2 at a concentration of 3 mg/mL (Invitrogen, Carlsbad, CA) or with the TLR3 antagonist CU CPT 4a at a concentration of 30 mg/mL (Tocris Bioscience, Ellisville, MOUK) for 3 hours and then stimulated with 600 mg HT exosomes.

Techniques: Immunofluorescence, Confocal Laser Scanning Microscopy, Flow Cytometry, Negative Control

Journal: The Journal of clinical endocrinology and metabolism

Article Title: Circulating Exosomes Activate Dendritic Cells and Induce Unbalanced CD4+ T Cell Differentiation in Hashimoto Thyroiditis.

doi: 10.1210/jc.2019-00273

Figure Lengend Snippet: Figure 4. HT exosomes activate the NF-kB signaling pathway in DCs, upregulate the expression of costimulatory molecules, and increase the release of IL-6. DCs were cocultured with HT exosomes or HC exosomes for 24 hours. To perform the inhibition experiments, DCs were pretreated with TLR2/3 inhibitors for 3 hours and then incubated with HT exosomes for 24 hours. (a) Representative stripe images and protein expression of MyD-88, TRIF, and p-P65 in the HC exosomes, HT exosomes (n 5 17 per group), HT exosomes 1 TLR2 inhibitor, and HT exosomes 1 TLR3 inhibitor groups (n 5 8 to 10 per group). PBS was added to the medium of a group without exosomes and inhibitors as a negative control (n 5 5). TLR2 inhibitor and TLR3 inhibitor groups were used as positive controls in the inhibition experiments (n 5 8 to 10 per group). The relative protein expression level was corrected to the GAPDH expression level. (b) Expression of IL-6 in the supernatants of the HC exosomes, HT exosomes (n 5 14 per group), HT exosomes 1 TLR2 inhibitor, and HT exosomes 1 TLR3 inhibitor groups (n 5 8 to 13 per group). PBS was added to the medium of a group without exosomes and inhibitors as a negative control (n 5 5). TLR2 inhibitor and TLR3 inhibitor groups were used as positive controls in the inhibition experiments (n 5 5). (c) Expression of the costimulatory molecules CD40, CD80, and CD83 on CD11c1DCs in the HC exosomes (n 5 11), HT exosomes (n 5 10), HT exosomes 1 TLR2 inhibitor, and HT exosomes 1 TLR3 inhibitor groups (n 5 6 to 8 per group). PBS was added to the medium of a group without exosomes and inhibitors as a negative control (n 5 5). TLR2 inhibitor and TLR3 inhibitor groups were used as positive controls in the inhibition experiments (n 5 6 to 4). Statistical analyses of multiple groups were performed by Kruskal-Wallis H-test or one-way ANOVA. *P , 0.05; **P , 0.01.

Article Snippet: To perform the inhibition test, according to the literature and the results of our preliminary experiments, DCs were preincubated with the TLR2 antagonist PAb hTLR2 at a concentration of 3 mg/mL (Invitrogen, Carlsbad, CA) or with the TLR3 antagonist CU CPT 4a at a concentration of 30 mg/mL (Tocris Bioscience, Ellisville, MOUK) for 3 hours and then stimulated with 600 mg HT exosomes.

Techniques: Expressing, Inhibition, Incubation, Negative Control

Journal: The Journal of clinical endocrinology and metabolism

Article Title: Circulating Exosomes Activate Dendritic Cells and Induce Unbalanced CD4+ T Cell Differentiation in Hashimoto Thyroiditis.

doi: 10.1210/jc.2019-00273

Figure Lengend Snippet: Figure 5. HT exosomes modulate the differentiation of Th1, Th17, and Treg cells and the secretion of cytokines. PBMCs were incubated with HT exosomes or HC exosomes for 1 to 4 days. To perform the inhibition experiments, PBMCs were pretreated for 3 hours with TLR2 or TLR3 inhibitors and then incubated with HT exosomes for 1 to 4 days. (a) Representative pictures of CD41IFN-g1Th1, CD41IL-17A1Th17A, and CD41CD251Foxp31 Treg cells as analyzed by flow cytometry in the inhibition experiments. (b) Frequency of CD41IFN-g1Th1, CD41IL- 17A1Th17A, and CD41CD251Foxp31 Treg cells in the HC exosomes, HT exosomes (n 5 30 per group), HT exosomes 1 TLR2 inhibitor, and HT exosomes 1 TLR3 inhibitor groups (n 5 10 to 12 per group). PBS was added to the medium of a group without exosomes and inhibitors as a negative control (n 5 3). TLR2 inhibitor and TLR3 inhibitor groups were used as a positive control in the inhibition experiments (n 5 5). (c) Levels of IFN-g, IL-17A, and IL-10 in the cell supernatants as determined by ELISA in the HC exosomes, HT exosomes (n 5 18 to 23 per group), HT exosomes 1 TLR2 inhibitor, and HT exosomes 1 TLR3 inhibitor groups (n 5 12 to 20 per group). PBS was added to the medium of a group without exosomes and inhibitors as a negative control (n 5 3). TLR2 inhibitor and TLR3 inhibitor groups were used as a positive control in the inhibition experiments (n 5 5). HT-exo 1 TLR2 inhibitor: HT exosomes 1 TLR2 inhibitor; HT-exo 1 TLR3 inhibitor: HT exosomes 1 TLR3 inhibitor. Statistical analyses of multiple groups were performed by Kruskal-Wallis H-test or one-way ANOVA. *P , 0.05; **P , 0.01.

Article Snippet: To perform the inhibition test, according to the literature and the results of our preliminary experiments, DCs were preincubated with the TLR2 antagonist PAb hTLR2 at a concentration of 3 mg/mL (Invitrogen, Carlsbad, CA) or with the TLR3 antagonist CU CPT 4a at a concentration of 30 mg/mL (Tocris Bioscience, Ellisville, MOUK) for 3 hours and then stimulated with 600 mg HT exosomes.

Techniques: Incubation, Inhibition, Flow Cytometry, Negative Control, Positive Control, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: Indoleamine 2,3-dioxygenase-regulated macrophages metabolic reprogramming rescues tacrolimus-induced nephrotoxicity

doi: 10.3389/fphar.2026.1784153

Figure Lengend Snippet: KYN supplementation alleviates TAC-induced energy metabolism disorder by restoring FAO. (A) Relative abundance of TCA cycle–related metabolites in the renal cortex (n = 5). (B) Relative abundance of glycolysis-related metabolites in the renal cortex (n = 5). (C) Schematic diagram of energy metabolism pathways based on metabolomic data, including the TCA cycle, glycolysis, and β-oxidation; red arrows indicate metabolites upregulated by TAC treatment, and blue arrows indicate metabolites downregulated by TAC treatment. (D) Heatmap of fatty acid metabolism–related compounds in the renal cortex (n = 5). (E,F) Seahorse analysis showing mitochondrial oxygen consumption rate (OCR) in renal cortical cells using glucose (E) or palmitic acid (F) as substrates (n = 4). (G) Quantification of basal and maximal OCR in renal cortical cells under glucose or palmitic acid substrate conditions (n = 4). (H) RT-qPCR analysis of FAO-related gene expression (MCAD, LCAD, CPT1, CPT2) in the renal cortex (n = 3). (I) Schematic diagram of mitochondrial fatty acid transport and β-oxidation pathways. (J) Western blot analysis of CPT1, CPT2, and CACT protein expression in the renal cortex, with β-tubulin as a loading control. (K) Densitometric quantification of CPT1, CPT2, and CACT protein bands from Western blots (n = 3). Data are presented as mean ± SD. Statistical significance is indicated as *P < 0.05, **P < 0.01, ***P < 0.001. Abbreviations: CIT, citrate; αKG, alpha-ketoglutarate; SUC, succinate; OAA, oxaloacetate; FUM, fumarate; MAL, malate; PYR, pyruvate; LAC, lactate; G6P, glucose-6-phosphate; F6P, fructose-6-phosphate; F1,6BP, fructose-1,6-bisphosphate; MCAD, medium-chain acyl-CoA dehydrogenase; LCAD, long-chain acyl-CoA dehydrogenase; CPT1/2, carnitine palmitoyltransferase 1/2; CACT, carnitine-acylcarnitine translocase; ACSL, acyl-CoA synthetase long-chain family member; OCR, oxygen consumption rate.

Article Snippet: Primary antibodies used in this study included: IDO1 (Proteintech, Cat# 13268-1-AP, 1:1000), CPT1 (ABclonal, Cat# A5307, 1:2000), CPT2 (Proteintech, Cat# 26555-1-AP, 1:8000), CACT (ABclonal, Cat# A13956, 1:400), β-tubulin (Proteintech, Cat# 80713-1-RR, 1:10,000), HRP-conjugated Goat Anti-Rabbit IgG (H + L) (ABclonal, Cat# SA00001-2, 1:10,000).

Techniques: Metabolomic, Quantitative RT-PCR, Gene Expression, Western Blot, Expressing, Control

Journal: JCI insight

Article Title: Combining antibiotic with anti-TLR2/TLR13 therapy prevents brain pathology in pneumococcal meningitis.

doi: 10.1172/jci.insight.165737

Figure Lengend Snippet: Figure 4. Dual neutralizing TLR2 mAb and CQ application in vitro abrogates Streptococcus pneumoniae–induced cytokine release by murine and human immune cells. (A and B) Murine BMDMs (blue boxes) and (C–F) human PBMCs (green boxes) were cultured and challenged with 1 × 106 CFU/mL viable S. pneumoniae (SP, except for controls, which received THY solution only). Four hours postinfection (p.i.), cells were treated with ceftriaxone. Two hours earlier (2 h), simultaneously (4 h), or 2 hours later (6 h p.i.), anti-TLR2 mAb (T2.5) and chloroquine (CQ) were added to cell cultures. In human PBMC cultures, the TLR8 inhibitor Cu-Cpt9a and dexamethasone (DEXA) were applied additionally where indicated. Cell culture supernatants were sampled 24 hours p.i., aliquoted, and stored until ELISA analysis. Supernatants were analyzed for IL-6 (A–D), IL-1β (E), and TNF (F). The experiments with BMDMs were conducted at least 2 times in triplicate, while those involving human PBMCs were performed at least 2 times in quadruplicate (for detailed information see Supporting Data Values file). The data for the placebo and T2.5 plus CQ (4 h) groups in A and B (as indicated by blue boxes with gray diagonal stripes in B) largely overlap. Data are depicted as median (line in box), 25%–75% percentile range (bounds of the box), and minimum and maximum (whiskers). Statis- tical test was 1-way ANOVA and subsequent Tukey’s post hoc test. ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05. NS, not significant.

Article Snippet: PBMCs and BMDMs were challenged in identical manners except for the treatment with dexamethasone (5 μg/ mL) and the TLR8 antagonist Cu-Cpt9a (10 μg/mL; Tocris) (66).

Techniques: In Vitro, Cell Culture, Enzyme-linked Immunosorbent Assay