Journal: Smart Molecules

Article Title: Endocytosis‐independent cytosolic entry of messenger RNA via fluorous bilayer zippering attenuating Toll‐like receptor signaling and enables ischemic tissue salvage

doi: 10.1002/smo2.70085

Figure Lengend Snippet: Biocompatibilities and immunogenicity of mRNA‐encapsulating polyplexes. (a) Cell viabilities of HUVECs upon 24 h incubation in presence of a variety of mRNA‐encapsulating polyplexes (mean ± s.d., n = 4, ** p < 0.01, student t test). (b) Hemolytic activities of sheep red blood cells upon 2 h incubation with varied concentrated delivery materials (*** p < 0.005; Student's t ‐test, mean ± s.d., n = 4). (c) confocal laser scanning microscopy (CLSM) measurement for assessment of overall cellular internalization of mRNA‐encapsulating polyplexes into RAW264.7 cells. (d) Interferon‐β (IFN‐β): at 4 h post incubation, the expression levels of inflammatory molecules were measured with qRT‐PCR. (e) Interleukin‐8 (IL‐8): at 4 h post incubation, the expression levels of inflammatory molecules were measured with qRT‐PCR. (** p < 0.01, *** p < 0.005; Student's t ‐test, n = 4).

Article Snippet: Culture supernatants (100 μL/well) were harvested and assayed for IFN‐α using the Mouse IFN‐α ELISA Kit (PBL Interferon Source) according to the manufacturer's instructions.

Techniques: Immunopeptidomics, Incubation, Confocal Laser Scanning Microscopy, Expressing, Quantitative RT-PCR

Journal: Smart Molecules

Article Title: Endocytosis‐independent cytosolic entry of messenger RNA via fluorous bilayer zippering attenuating Toll‐like receptor signaling and enables ischemic tissue salvage

doi: 10.1002/smo2.70085

Figure Lengend Snippet: Revascularization in hindlimbs by local dosage of mVEGF‐encapsulating polyplexes. (a) Therapeutic scheme. (b) Anatomy of the established hindlimb ischemia model. Ligations were made in the femoral artery at the proximal and distal sites. (c) Angiogenesis in mouse hindlimbs post ligation. (d) Visualization of blood flow by Laser Speckle Flowgraphy on Day 28 post‐dosage of mVEGF therapeutics (mVEGF: 10 μg). The magnified inset images captured by intravital confocal laser scanning microscopy (CLSM), revealing vasculature details by intravenous dosage of FITC‐dextran (MW: 10 kDa). (e) Estimation of blood perfusion volume based on quantification by laser speckle flowgraphy on Day 28 post‐dosage of mVEGF therapeutics (mVEGF: 10 μg). The data were represented as the mean ± standard deviations (s.d.) ( n = 5). (* p < 0.05, ** p < 0.01, student t test). (f) Quantification of the expressed VEGF protein on day 4 post dosage by ELISA.

Article Snippet: Culture supernatants (100 μL/well) were harvested and assayed for IFN‐α using the Mouse IFN‐α ELISA Kit (PBL Interferon Source) according to the manufacturer's instructions.

Techniques: Ligation, Confocal Laser Scanning Microscopy, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cardiovascular Development and Disease

Article Title: Study on the Role and Mechanism of TOPORS in Regulating Aortic Dissection by Mediating SUMOylation

doi: 10.3390/jcdd13030110

Figure Lengend Snippet: Inhibition of TOPORS mitigated aortic dissection (AD) in mice. ( A ): Ultrasound imaging of the mouse thoracic aorta depicting changes in aortic diameter (scale bar = 1 mm); ( B ): Gross anatomical images of the thoracic aorta demonstrating morphological variations (scale bar = 2 mm); ( C ): Representative transverse sections of aortic tissues from AD mice stained with hematoxylin and eosin (H&E) for structural analysis and elastica van Gieson (EVG) for visualization of elastic fibers; ( D , E ): Expression levels of TOPORS in aorta tissues determined by qRT-PCR for mRNA ( D ) and Western blot ( E ) for protein; F-I: Enzyme-linked immunosorbent assay (ELISA) quantification of pro-inflammatory factors in aortic tissues: ( F ) interferon-α ( IFN- α), ( G ) interleukin-6 ( IL-6 ), ( H ) tumor necrosis factor-α ( TNF-α ), and ( I ) interleukin-1β (IL-1β). Data are presented as mean ± standard deviation (SD) ( n = 10). Significance levels are denoted as * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, where “ns” indicates no significant difference.

Article Snippet: Specifically, the following ELISA kits were employed: Mouse TNF-α Elisa Kit (H052-1-2), Mouse IFN-α Elisa Kit (H023-1-1), Mouse IL-6 Elisa Kit (H007-1-2), and Mouse IL-1β Elisa Kit (H002-1-2), all purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Inhibition, Dissection, Imaging, Staining, Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Journal of Cardiovascular Development and Disease

Article Title: Study on the Role and Mechanism of TOPORS in Regulating Aortic Dissection by Mediating SUMOylation

doi: 10.3390/jcdd13030110

Figure Lengend Snippet: Effects of TOPORS on Ang-II-induced vascular smooth muscle cells (VSMCs). ( A ): Ang-II treatment leads to a dose-dependent downregulation of mRNA expression for calponin and SM22α , which are critical markers of the contractile VSMC phenotype; ( B ): The mRNA expression level of TOPORS in VSMC lines with TOPORS knockout (si TOPORS ) or overexpression (oe TOPORS ) was determined by qRT-PCR; ( C ): Regulation of TOPORS knockout or overexpression on the viability of Ang-II-stimulated VSMCs was assessed by CCK-8 assay; ( D , E ): Regulation of TOPORS knockout or overexpression on the apoptosis of Ang-II-stimulated VSMCs was evaluated by flow cytometry analysis ( D ) and TUNEL staining ( E ); ( F – I ): Regulation of TOPORS knockout or overexpression on the secretion of pro-inflammatory cytokines in VSMC lines were determined by ELISA: interleukin-6 ( IL-6 ) ( F ), interferon-α ( IFN- α) ( G ), tumor necrosis factor-α ( TNF-α ) ( H ), and interleukin-1β ( IL-1β ) ( I ). Data are presented as mean ± standard deviation (SD) ( n = 3). Significance levels are denoted as * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, where “ns” represents no significant difference.

Article Snippet: Specifically, the following ELISA kits were employed: Mouse TNF-α Elisa Kit (H052-1-2), Mouse IFN-α Elisa Kit (H023-1-1), Mouse IL-6 Elisa Kit (H007-1-2), and Mouse IL-1β Elisa Kit (H002-1-2), all purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Expressing, Knock-Out, Over Expression, Quantitative RT-PCR, CCK-8 Assay, Flow Cytometry, TUNEL Assay, Staining, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Journal of Cardiovascular Development and Disease

Article Title: Study on the Role and Mechanism of TOPORS in Regulating Aortic Dissection by Mediating SUMOylation

doi: 10.3390/jcdd13030110

Figure Lengend Snippet: Effects of TOPORS on PI3K/AKT and p53 signaling pathways in VSMC model of aortic dissection ( A ). ( A – C ): the mRNA expression profiles of AKT ( A ), PI3K ( B ), and p53 ( C ) quantified by qRT-PCR. ( D ): Western blot analysis of protein expression, including AKT, p-AKT, PI3K, p-PI3K, and p53 . ( E , F ): The apoptosis of AD model VSMC after PFT-ɑ treatment was evaluated by TUNEL staining ( E ) and flow cytometry analysis ( F ); ( G – J ): ELISA quantification of pro-inflammatory cytokines after PFT-ɑ treatment: tumor necrosis factor-α ( TNF-α ) ( G ), interleukin-6 ( IL-6 ) ( H ), interleukin-1β ( IL-1β ) ( I ), and interferon-α ( IFN- α) ( J ). Data are presented as mean ± standard deviation (SD) ( n = 3). Significance levels are denoted as * p < 0.05, *** p < 0.001, **** p < 0.0001, where “ns” represents no significant difference. ( K ): The SUMOylation level of the p53 gene was detected by co-immunoprecipitation (Co-IP) in the model-si TOPORS group and model-oe TOPORS group.

Article Snippet: Specifically, the following ELISA kits were employed: Mouse TNF-α Elisa Kit (H052-1-2), Mouse IFN-α Elisa Kit (H023-1-1), Mouse IL-6 Elisa Kit (H007-1-2), and Mouse IL-1β Elisa Kit (H002-1-2), all purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Protein-Protein interactions, Dissection, Expressing, Quantitative RT-PCR, Western Blot, TUNEL Assay, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Standard Deviation, Immunoprecipitation, Co-Immunoprecipitation Assay

Journal: Microorganisms

Article Title: Heat-Killed Enterococcus faecalis EF-2001 Promotes Systemic Th1-Skewed Immune Activation Without Detectable Reduction of Influenza Viral Load in Mice

doi: 10.3390/microorganisms14020316

Figure Lengend Snippet: Lung viral titers (TCID 50 ) and IFN-α concentrations on day 2 post-infection. No significant differences were observed between EF-2001 treated groups and controls, indicating that systemic Th1 priming did not translate to immediate viral reduction in the lungs at 48 h.

Article Snippet: Lung IFN-α levels were determined from the same supernatants using the VeriKine Mouse IFN-α ELISA kit (PBL Assay Science, Tokyo, Japan), with absorbance read at 450 nm on a SpectraMax ABSPlus microplate reader (Molecular Devices, Tokyo, Japan).

Techniques: Infection