Journal: Pharmaceutics

Article Title: Design and Evaluation of Autophagy-Inducing Particles for the Treatment of Abnormal Lipid Accumulation

doi: 10.3390/pharmaceutics14071379

Figure Lengend Snippet: In vivo and organ DiR fluorescence imaging of genetically obese ob/ob mice following a single IV injection (120 µL) of soluble fluorophore or fluorescent particles (0.25% w/v ). ( a ) Autophagy-inducing fluorescent particles (NP DiR T-B) and plain fluorescent particles (NP DiR) accumulate in the liver over the course of 72 h. Images are representative of scans from 3–4 mice per group. ( b ) Quantification of whole-body fluorescence up to 72 h post a single IV injection. The percentage values reported are calculated from the 5 min timepoint, when the maximal fluorescence was observed, and are reported as mean ± s.e.m. Results were analyzed using one-way ANOVA followed by Sidak’s multiple comparison test, which revealed no significant difference between the NP DiR and the NP DiR T-B group for all timepoints. ( c , d ) Organ biodistribution of fluorescent NPs 24 h after a single intravenous (IV) injection in ob/ob mice. n = 3–4 mice per group. Ex vivo DiR fluorescence imaging on main organs (liver, spleen, pancreas and kidneys) ( c ) and quantification of DiR fluorescence ( d ). The percentage values reported are calculated from total organ fluorescence and presented as mean ± s.e.m ( d ).

Article Snippet: Mice were immediately injected in the retro-orbital vein with 120 µL of either free fluorophore (Xenolight DiR-Fluorescent Dye, PerkinElmer, Waltham, MA, USA, 125964), fluorescent nanoparticles (NP DiR, Near Infra-Red Fluorescent i-Particles ® , Adjuvatis, France) or fluorescent Tat-Beclin nanoparticles (NP DiR T-B) of 0.25% w/v solid content.

Techniques: In Vivo, Fluorescence, Imaging, IV Injection, Comparison, Ex Vivo

Journal: Frontiers in Pharmacology

Article Title: Differential Mechanisms of Action and Efficacy of Vitamin E Components in Antioxidant Cytoprotection of Human Retinal Pigment Epithelium

doi: 10.3389/fphar.2021.798938

Figure Lengend Snippet: Effect of 24-h αT, γT and δT and αTr exposure on RPE actin dynamics after subsequent tBHP-induced oxidative stress. RPE cells were exposed to a vehicle control (VC) αT, γT or δT or αTr for 24 h prior to tBHP. (A) tBHP reduced phalloidin fluorescence (red) and nuclear area (DAPI, blue fluorescence). A reduction in filamentous actin (phalloidin (red) fluorescence) is suggestive of actin destabilization observed in cell death resembling apoptosis. Nuclear size (blue) is reduced during cell death, particularly during apoptosis. Images were acquired at 40X magnification; scale bar is 100 μm. (B) A close up image of tBHP-mediated reduction in filamentous actin labeling by phalloidin and reduction in nucleus size caused by tBHP exposure. Scale bar is 25 μm. (C) Graphical data showing that exposure of cells to αT and γT, and to a lesser extent δT, prevented this effect in hTERT-RPE cells. αTr, however, could not prevent the reduction of phalloidin staining and nuclear size. (D) Exposure of cells to αT and δT led to an increase in nuclear size. Exposure of cells to tBHP led to a reduction in nucleus size indicative of pyknosis. The exposure of cells to αT, γT and δT prevented the tBHP-mediated reduction in nuclear size, while exposure of cells to αTr did not. ** p ≤ 0.01 with tBHP versus the same condition without tBHP; ## p ≤ 0.01 with tocopherol versus without tocopherol. Microfluorimetric analyses are from an average of three images. One-way ANOVA analysis with post hoc Bonferroni correction was performed to determine differences between groups.

Article Snippet: For cell staining of fixed cells, nuclei were labeled with dilute 4′,6-diamidine-2′-phenylindole dihydrochloride (DAPI) stain (Millipore Sigma) and fluorescent dye 594 nm-labeled phalloidin (phalloidin-594; Abnova, Taipei City, Taiwan).

Techniques: Control, Fluorescence, Labeling, Staining

Journal: Respiratory Research

Article Title: Intranasal delivery of R8-modified circNFXL1 liposomes ameliorates Su5416-induced pulmonary arterial hypertension in C57BL/6 mice

doi: 10.1186/s12931-025-03203-y

Figure Lengend Snippet: Synthesis and Characterization of R8-PEG2000 Liposomal circNFXL1 for Targeted Pulmonary Delivery. A Schematic representation of the preparation of R8-PEG2000-modified liposomal circNFXL1 (R8-circNFXL1). B Transmission electron microscopy (TEM) images of Lip-circNFXL1 ( left ) and R8-Lip-circNFXL1 ( right ), show uniform, spherical structures with well-defined edges, indicative of stable vesicle formation. C Dynamic light scattering (DLS) analysis of particle size distribution. D The average size of Lip-circNFXL1 is 228.3 ± 5.9 nm, while R8-Lip-circNFXL1 exhibits an average size of 254.5 ± 5.1 nm. E Polydispersity index (PDI) values for Lip-circNFXL1 and R8-Lip-circNFXL1. F Zeta potential measurements show a significant increase in the negative surface charge for R8-Lip-circNFXL1 compared to Lip-circNFXL1. Data are presented as mean ± SEM. Statistical comparisons were conducted using Student’s t -test for pairwise comparisons. n = 6 per group. ** p < 0.01 compared to the Lip-circNFXL1 group

Article Snippet: To assess the expression and localization of circNFXL1 and miR-29b, we employed RNA-fluorescence in situ hybridization (RNA-FISH) using a cyanine dye-labeled red fluorescent probe for circNFXL1 and a green FAM-labeled fluorescent probe for miR-29b (GenePharma Co., Ltd., Shanghai, China).

Techniques: Modification, Transmission Assay, Electron Microscopy, Zeta Potential Analyzer

Journal: Respiratory Research

Article Title: Intranasal delivery of R8-modified circNFXL1 liposomes ameliorates Su5416-induced pulmonary arterial hypertension in C57BL/6 mice

doi: 10.1186/s12931-025-03203-y

Figure Lengend Snippet: Encapsulation Efficiency and Enzymatic Stability of Liposomal circNFXL1 Formulations. Encapsulation efficiency percentages of Lip-circNFXL1 ( A ) and R8-circNFXL1 ( B ) at various concentrations (0.1, 0.5, 1.0, and 2.0 mg/mL). Agarose gel electrophoresis results show the integrity of circNFXL1 ( C ) within Lip-circNFXL1 and R8-circNFXL1 ( D ) formulations after exposure to EcoRI and BamHI enzymes. Zeta potential measurements (mV) of Lip-circNFXL1 ( E ) and R8-circNFXL1 ( F ) formulations, illustrating electrostatic stability. Data are presented as mean ± SEM. Statistical comparisons were conducted using one-way ANOVA, followed by Bonferroni’s post hoc test for multiple group comparisons. ** p < 0.01 compared to the 0.1 mg/mL group. A , B : n = 3 independent experiments; E , F : n = 7 per group

Article Snippet: To assess the expression and localization of circNFXL1 and miR-29b, we employed RNA-fluorescence in situ hybridization (RNA-FISH) using a cyanine dye-labeled red fluorescent probe for circNFXL1 and a green FAM-labeled fluorescent probe for miR-29b (GenePharma Co., Ltd., Shanghai, China).

Techniques: Encapsulation, Agarose Gel Electrophoresis, Zeta Potential Analyzer

Journal: Respiratory Research

Article Title: Intranasal delivery of R8-modified circNFXL1 liposomes ameliorates Su5416-induced pulmonary arterial hypertension in C57BL/6 mice

doi: 10.1186/s12931-025-03203-y

Figure Lengend Snippet: Biocompatibility and Safety Profile of Liposomal circNFXL1 Formulations. A Hemolysis assays demonstrating the compatibility of Lip-circNFXL1 and R8-circNFXL1 with red blood cells. Quantitative analysis of hemolysis percentages for Lip-circNFXL1 (n = 3 independent experiments) ( B ) and R8-circNFXL1 (n = 3 independent experiments) ( C ) formulations at different concentrations. Negative control: PBS; Positive control: H 2 O. D Histopathological evaluation of major organs (heart, liver, spleen, lung, and kidney) from mice treated with Lip-circNFXL1 and R8-circNFXL1 compared to untreated controls

Article Snippet: To assess the expression and localization of circNFXL1 and miR-29b, we employed RNA-fluorescence in situ hybridization (RNA-FISH) using a cyanine dye-labeled red fluorescent probe for circNFXL1 and a green FAM-labeled fluorescent probe for miR-29b (GenePharma Co., Ltd., Shanghai, China).

Techniques: Negative Control, Positive Control

Journal: Respiratory Research

Article Title: Intranasal delivery of R8-modified circNFXL1 liposomes ameliorates Su5416-induced pulmonary arterial hypertension in C57BL/6 mice

doi: 10.1186/s12931-025-03203-y

Figure Lengend Snippet: In vivo Biodistribution of Liposomal circNFXL1 via Intravenous and Intranasal Administration in Mice. Representative fluorescence images showing the biodistribution of R8-circNFXL1, R8-PLC5-DID, and free DID dye in C57BL/6 mice at various time points (1, 6, 12, and 24 h) following A intravenous and B intranasal administration

Article Snippet: To assess the expression and localization of circNFXL1 and miR-29b, we employed RNA-fluorescence in situ hybridization (RNA-FISH) using a cyanine dye-labeled red fluorescent probe for circNFXL1 and a green FAM-labeled fluorescent probe for miR-29b (GenePharma Co., Ltd., Shanghai, China).

Techniques: In Vivo, Fluorescence

Journal: Respiratory Research

Article Title: Intranasal delivery of R8-modified circNFXL1 liposomes ameliorates Su5416-induced pulmonary arterial hypertension in C57BL/6 mice

doi: 10.1186/s12931-025-03203-y

Figure Lengend Snippet: Ex vivo Biodistribution of Liposomal circNFXL1 in Mice Following Intravenous and Intranasal Administration. Representative fluorescence images showing the ex vivo biodistribution of R8-circNFXL1, R8-PLC5-DID, and free DID dye in major organs (brain, heart, liver, spleen, lungs, and kidneys) of C57BL/6 mice at 24 h post-administration following A intravenous and B intranasal administration

Article Snippet: To assess the expression and localization of circNFXL1 and miR-29b, we employed RNA-fluorescence in situ hybridization (RNA-FISH) using a cyanine dye-labeled red fluorescent probe for circNFXL1 and a green FAM-labeled fluorescent probe for miR-29b (GenePharma Co., Ltd., Shanghai, China).

Techniques: Ex Vivo, Fluorescence

Journal: Respiratory Research

Article Title: Intranasal delivery of R8-modified circNFXL1 liposomes ameliorates Su5416-induced pulmonary arterial hypertension in C57BL/6 mice

doi: 10.1186/s12931-025-03203-y

Figure Lengend Snippet: Therapeutic Efficacy of R8-circNFXL1 in a Mouse Model of Pulmonary Arterial Hypertension (PAH). A Experimental timeline illustrating PAH induction in mice using hypoxia and Su5416, followed by treatment with R8-circNFXL1. B Representative echocardiographic images and quantitative analyses of pulmonary and right ventricular (RV) function across different groups. B Doppler echocardiography depicting pulmonary artery acceleration time (PAT) and ejection time (PET), with the control, PAH, and R8-circNFXL1 treatment groups shown ( upper ). Representative M-mode echocardiographic images showing the right ventricular structure in each group ( lower ). C Quantitative measurement of RV/LV+S ratio (Fulton index), a marker of RV hypertrophy, in control, PAH, and R8-circNFXL1 treatment groups. Quantification of PAT ( D ), PAT/PET ratio ( E ), PAVTI (pulmonary artery velocity–time integral) ( F ), PVmax (peak velocity) ( G ), TAPSE (tricuspid annular plane systolic excursion) ( H ) and CO (cardiac output) ( I ) in control, PAH, and R8-circNFXL1 treatment groups. Data are presented as mean ± SEM. Statistical comparisons were performed using one-way ANOVA, followed by Bonferroni’s post hoc test for multiple group comparisons. ** p < 0.01, *** p < 0.001 compared to Nor group, # p < 0.05, ## p < 0.01 compared to PAH group (n = 3–6 per group)

Article Snippet: To assess the expression and localization of circNFXL1 and miR-29b, we employed RNA-fluorescence in situ hybridization (RNA-FISH) using a cyanine dye-labeled red fluorescent probe for circNFXL1 and a green FAM-labeled fluorescent probe for miR-29b (GenePharma Co., Ltd., Shanghai, China).

Techniques: Drug discovery, Control, Marker

Journal: Respiratory Research

Article Title: Intranasal delivery of R8-modified circNFXL1 liposomes ameliorates Su5416-induced pulmonary arterial hypertension in C57BL/6 mice

doi: 10.1186/s12931-025-03203-y

Figure Lengend Snippet: Effects of R8-circNFXL1 on Pulmonary Vascular Remodeling and Right Ventricular Hypertrophy in a PAH Mouse Model. A Representative HE staining of lung sections illustrating the heart morphology across experimental groups ( upper ). Higher magnification HE images of lung sections highlighting detailed structural differences in vascular morphology and remodeling severity across groups ( lower ). Quantification of length of RV ( B ) and vessel wall thickness of pulmonary artery ( C ) across groups. D Immunofluorescence staining for α-SMA ( red ), circNFXL1 ( green ), and nuclei ( blue ) in lung sections across groups. E Quantification of the expression of circNFXL1 across groups. qRT-PCR analysis of circNFXL1 ( F ), miR-29b ( G ), and Kcnb1 ( H ) expression levels in pulmonary artery tissues. I Western blot analysis and quantification of KCNB1 protein levels. Data are presented as mean ± SEM. Statistical comparisons were conducted using one-way ANOVA, followed by Bonferroni’s post hoc test for multiple group comparisons. ** p < 0.01, *** p < 0.001 compared to Nor group, # p < 0.05, ## p < 0.01, ### p < 0.001 compared to PAH group (n = 4–9 per group)

Article Snippet: To assess the expression and localization of circNFXL1 and miR-29b, we employed RNA-fluorescence in situ hybridization (RNA-FISH) using a cyanine dye-labeled red fluorescent probe for circNFXL1 and a green FAM-labeled fluorescent probe for miR-29b (GenePharma Co., Ltd., Shanghai, China).

Techniques: Staining, Immunofluorescence, Expressing, Quantitative RT-PCR, Western Blot

Journal: Respiratory Research

Article Title: Intranasal delivery of R8-modified circNFXL1 liposomes ameliorates Su5416-induced pulmonary arterial hypertension in C57BL/6 mice

doi: 10.1186/s12931-025-03203-y

Figure Lengend Snippet: Modulation of the miR-29b/Kcnb1 Axis by R8-circNFXL1 in Pulmonary Arterial Hypertension (PAH). A Immunofluorescence staining of lung tissues showing circNFXL1 ( green ), miR-29b ( red ), Kcnb1 ( purple ), and DAPI ( blue ) across different experimental groups: Normal (Nor), PAH, and PAH with R8-circNFXL1 treatment. Quantitative analysis of fluorescence intensity for circNFXL1 ( B ), miR-29b ( C ), and KCNB1 ( D ) in pulmonary artery tissue sections across groups. Data are presented as mean ± SEM. Statistical comparisons were conducted using one-way ANOVA, followed by Bonferroni’s post hoc test for multiple group comparisons. * p < 0.05, ** p < 0.01, *** p < 0.001 compared to Nor group, # p < 0.05, ## p < 0.01, ### p < 0.001 compared to PAH group (n = 3 per group)

Article Snippet: To assess the expression and localization of circNFXL1 and miR-29b, we employed RNA-fluorescence in situ hybridization (RNA-FISH) using a cyanine dye-labeled red fluorescent probe for circNFXL1 and a green FAM-labeled fluorescent probe for miR-29b (GenePharma Co., Ltd., Shanghai, China).

Techniques: Immunofluorescence, Staining, Fluorescence