Journal: International Journal of Nanomedicine

Article Title: Glucose-responsive mesoporous silica nanoparticles to generation of hydrogen peroxide for synergistic cancer starvation and chemistry therapy

doi: 10.2147/IJN.S195900

Figure Lengend Snippet: ( A ) The morphological changes of HepG2 cells detected with dual staining of Hoechst 33,342/PI and captured by fluorescence microscope (magnification 200x). Arrows indicate (1) viable cells with normal nuclei; (2) live cells with apoptotic nuclei; (3) dead cells with normal nuclei; and (4) dead cells with apoptotic nuclei. ( B ) Apoptosis rate of HepG2 cells was determined by Annexin V-FITC/PI staining. ( C ) Apoptosis rate of HepG2 cells was analyzed by Annexin V-FITC/PI staining after treatment with MSNs-GOx/PLL/HA in in different concentrations (50, 100, 200, 400, 800 μg/mL) of glucose. ( D ) Change in mitochondrial membrane potential ΔΨm of HepG2 cells treated with various preparations for 8 hours analyzed by flow cytometry. Significant difference from control: * P <0.05, ** P <0.01, *** P <0.001. ( E ) Effects of treatment with different preparations on the cell cycle of HepG2 cells. ( F ) The percent of cell cycle distribution after treatment with various formulations for 24 hours. ( G ) Plasma concentration-time curves of taxol and PTX-loaded nanoparticles in rats after intravenous administration of 2.5 mg/kg PTX (mean ±SD, N=6). Abbreviations: PI, propidium iodide; FITC, fluorescein isothiocyanate; MSNs, mesoporous silica nanoparticles; GOx, glucose oxidase; PLL, poly (L-lysine); HA, hyaluronic acid.

Article Snippet: The apoptosis-inducing capability of nanoparticles was further evaluated by Annexin V-FITC/PI Apoptosis Detection Kit (Beyotime).

Techniques: Staining, Fluorescence, Microscopy, Membrane, Flow Cytometry, Control, Clinical Proteomics, Concentration Assay

Journal: Advanced Materials (Deerfield Beach, Fla.)

Article Title: Unlocking Intracellular Protein Delivery by Harnessing Polymersomes Synthesized at Microliter Volumes using Photo‐PISA

doi: 10.1002/adma.202408000

Figure Lengend Snippet: In vitro activation of BMDCs and antigen cross‐presentation mediated by OVA‐loaded pH‐responsive PSomes. A) Schematic illustration of OVA‐loaded pH‐responsive PSomes enhancing maturation and antigen cross‐presentation in dendritic cells. Characterization of surface markers B) CD80, C) CD86, D) MHC class II, and E) SIINKEFL (OVA (257‐264)) bound to MHC class I by immunoflow cytometry analysis under various treatments in CD11c + cells. Cells were stained with antibodies against CD11c as the DC marker, CD80, CD86, and MHC class II as DC maturation markers as well as SIINKEFL‐H2K b to measure OVA antigen cross‐presentation. Data presented as mean ± SD ( n = 3, technical replicates). Statistical significance was determined using one‐way ANOVA with Dunnett's multiple comparisons test. **** p < 0.0001; ns not significant.

Article Snippet: A useful tool in this regard to study the loading of macromolecules on a single‐particle basis is nanoparticle flow cytometry (NanoFCM), which is a highly sensitive form of conventional flow cytometry but for nanoparticles.

Techniques: In Vitro, Activation Assay, Cytometry, Staining, Marker

Journal: Redox Biology

Article Title: Specific delivery of delta-5-desaturase siRNA via RNA nanoparticles supplemented with dihomo-γ-linolenic acid for colon cancer suppression

doi: 10.1016/j.redox.2018.101085

Figure Lengend Snippet: Construction of tumor-targeting 3WJ nanoparticle. (A) Illustration for the design of a functional 3WJ RNA nanoparticle. (B) Dynamic light scattering characterization showing the hydrodynamic size of RNA nanoparticles. (C) Atomic force microscopy image of extended 3WJ-nanoparticles. (D) Cell-binding test via flow cytometry in HCA-7/C29 cells after being incubated with 100 nM 3WJ-Alexa 647 or 3WJ-EpCAM-Alexa 647 (in 100 µL PBS). Cells incubated with PBS only (without nanoparticles) served as controls. (E) Confocal microscopy analysis of 3WJ-EpCAM-Alexa 647 particles entering HCA-7/C29 cells. Cell skeletons were stained with Alexa 488 and cell nuclei with DAPI. (F) qRT-PCR analysis for D5D gene knockdown efficiency in HCA-7/C29 cells after treatment with 10 nM of 3WJ nanoparticles with different compositions. Data represent mean ± SD with three separate experiments. *: significant difference with p < 0.05.

Article Snippet: The apparent hydrodynamic sizes of the assembled RNA nanoparticles (40 μM in 1 ×Tris buffer) were measured by a Zetasizer nano-ZS (Malvern Instrument, LTD) at 25 °C.

Techniques: Functional Assay, Microscopy, Binding Assay, Flow Cytometry, Incubation, Confocal Microscopy, Staining, Quantitative RT-PCR, Knockdown

Journal: Redox Biology

Article Title: Specific delivery of delta-5-desaturase siRNA via RNA nanoparticles supplemented with dihomo-γ-linolenic acid for colon cancer suppression

doi: 10.1016/j.redox.2018.101085

Figure Lengend Snippet: 3WJ nanoparticles and DGLA supplementation suppress xenograft tumor growth. (A) In vivo biodistribution analysis of 3WJ-EpCAM-Alexa 647 particles in organs and tumor tissues from mice bearing HCA-7 tumors at 8-h post-injection. (B) Ultrasound images of tumor growth in mice before and after 4-week treatments. Note, week 0 indicates the starting point of treatment, which is two weeks after cancer cell implantation. (C) Relative tumor volume measured twice a week using a digital caliper during the treatment. Data represent mean±SD with six tumor samples. *: significant difference with p < 0.05. Insert: photos of tumor tissues at the end of treatment.

Article Snippet: The apparent hydrodynamic sizes of the assembled RNA nanoparticles (40 μM in 1 ×Tris buffer) were measured by a Zetasizer nano-ZS (Malvern Instrument, LTD) at 25 °C.

Techniques: In Vivo, Injection

Journal: Redox Biology

Article Title: Specific delivery of delta-5-desaturase siRNA via RNA nanoparticles supplemented with dihomo-γ-linolenic acid for colon cancer suppression

doi: 10.1016/j.redox.2018.101085

Figure Lengend Snippet: 3WJ nanoparticles and DGLA supplementation promote 8-HOA formation in xenograft tumors. (A) Immunofluorescence analysis of D5D expression levels in tumor tissues. (B) LC/MS quantification of DGLA levels from tumor tissues. (C) LC/MS quantification of AA levels from tumor tissues. (D) DGLA/AA ratio from tumor tissues. (E) GC/MS quantification of 8-HOA from tumor tissues. All the quantification data represent mean±SD with six tumor samples. *: significant difference with p < 0.05.

Article Snippet: The apparent hydrodynamic sizes of the assembled RNA nanoparticles (40 μM in 1 ×Tris buffer) were measured by a Zetasizer nano-ZS (Malvern Instrument, LTD) at 25 °C.

Techniques: Immunofluorescence, Expressing, Liquid Chromatography with Mass Spectroscopy, Gas Chromatography-Mass Spectrometry

Journal: Journal of Asthma and Allergy

Article Title: MALAT1 Induces Food Allergy by Promoting Release of IL-6 from Dendritic Cells and Suppressing the Immunomodulatory Function of Tregs

doi: 10.2147/JAA.S341742

Figure Lengend Snippet: Effect of silencing or overexpression of MALAT1 on the maturation and IL-6 secretion of DCs. ( A ) RT-qPCR quantification of ZFP36 expression in mouse-sourced DCs. ( B ) Western blot assay to measure ZFP36 protein expression in mouse-sourced DCs. ( C ) ELISA to measure IL-12 levels in DC supernatant. ( D) ELISA to measure IL-6 levels in DC supernatant. ( E ) Expression of CD83 and CD86 in DCs determined by flow cytometry (in panels ( A – E ), n = 10, * p < 0.05 vs the control group, # p < 0.05 vs the model + sh-NC group). ( F ) Overexpression efficiency of oe-MALAT1 determined by RT-qPCR (* p < 0.05 vs the oe-NC group. ( G ) Expression of MALAT1 and ZFP36 in DCs determined by RT-qPCR. ( H ) ZFP36 protein expression in DCs quantified by Western blot analysis. ( I ) IL-12 level in DC supernatant by ELISA. ( J ) IL-6 level in DC supernatant by ELISA. ( K ) Expression of CD83 and CD86 in DCs by flow cytometry (in panels ( G – K ), * p < 0.05 vs the DCs group, # p < 0.05 vs the DCs + ox-LDL + oe-NC group). All cellular experiments were repeated 3 times independently.

Article Snippet: Protein separation was done using sodium dodecyl sulfate polyacrylamide gel electrophoresis and the proteins were transferred onto a polyvinylidene fluoride membrane (Millipore, Burlington, MA, USA) by wet transfer method, followed by blocking at ambient temperature with 5% bovine serum albumin for 1 h. Subsequently, diluted primary antibodies against β-actin (4970, 1: 5000, Cell Signaling Technology, Dancers, MA, USA), ZFP36 (ab204922, 1: 5000, Abcam, Cambridge, UK), IL-10 (ab189392, 1: 1000, Abcam), Foxp3 (ab215206, 1: 1000, Abcam), and TGF-β (ab215715, 1: 1000, Abcam) were allowed to incubate with the membrane overnight at 4°C.

Techniques: Over Expression, Quantitative RT-PCR, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Control

Journal: Journal of Asthma and Allergy

Article Title: MALAT1 Induces Food Allergy by Promoting Release of IL-6 from Dendritic Cells and Suppressing the Immunomodulatory Function of Tregs

doi: 10.2147/JAA.S341742

Figure Lengend Snippet: Effect of overexpressed ZFP36 on IL-6 secretion in DCs. ( A ) Transfection efficiency of oe-ZFP36 in DCs determined by RT-qPCR (* p < 0.05 vs the oe-NC group). ( B ) ZFP36 expression in DCs determined by RT-qPCR. ( C ) ZFP36 protein band in DCs detected by Western blot analysis. ( D ) ZFP36 protein level in DCs determined by Western blot analysis. ( E ) ELISA to measure IL-12 expression levels in DC supernatant. ( F ) ELISA to measure IL-6 expression in DC supernatant. ( G ) Expression of CD83 and CD86 in DCs measured by flow cytometry. ( H ) CD83 and CD86 expression in DCs measured by flow cytometry (in panels ( B – H ), * p < 0.05 vs the DCs group, # p < 0.05 vs the DCs + ox-LDL + oe-NC group). All cellular experiments were repeated 3 times independently.

Article Snippet: Protein separation was done using sodium dodecyl sulfate polyacrylamide gel electrophoresis and the proteins were transferred onto a polyvinylidene fluoride membrane (Millipore, Burlington, MA, USA) by wet transfer method, followed by blocking at ambient temperature with 5% bovine serum albumin for 1 h. Subsequently, diluted primary antibodies against β-actin (4970, 1: 5000, Cell Signaling Technology, Dancers, MA, USA), ZFP36 (ab204922, 1: 5000, Abcam, Cambridge, UK), IL-10 (ab189392, 1: 1000, Abcam), Foxp3 (ab215206, 1: 1000, Abcam), and TGF-β (ab215715, 1: 1000, Abcam) were allowed to incubate with the membrane overnight at 4°C.

Techniques: Transfection, Quantitative RT-PCR, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Journal of Asthma and Allergy

Article Title: MALAT1 Induces Food Allergy by Promoting Release of IL-6 from Dendritic Cells and Suppressing the Immunomodulatory Function of Tregs

doi: 10.2147/JAA.S341742

Figure Lengend Snippet: MALAT1 promotes IL-6 secretion from DCs and inhibits Treg immunomodulatory function by downregulating ZFP36 expression to facilitate FA. ( A ) RT-qPCR to determine MALAT1 expression levels in FA mouse jejunal tissue samples. ( B ) RT-qPCR to determine ZFP36 expression levels in FA mouse jejunal tissue samples. ( C ) Number of mice with diarrhea. ( D ) Relative rectal temperature of mice. ( E ) ELISA to measure serum-specific IgE and IgG1 levels in mice. ( F ) ELISA to measure IL-12 and IL-6 expression in DC supernatant. ( G ) Determination of CD83 and CD86 expression levels in DCs by flow cytometry. n = 10, * p < 0.05 vs the control group, # p < 0.05 vs the model and model + oe-NC group. All cellular experiments were repeated 3 times independently.

Article Snippet: Protein separation was done using sodium dodecyl sulfate polyacrylamide gel electrophoresis and the proteins were transferred onto a polyvinylidene fluoride membrane (Millipore, Burlington, MA, USA) by wet transfer method, followed by blocking at ambient temperature with 5% bovine serum albumin for 1 h. Subsequently, diluted primary antibodies against β-actin (4970, 1: 5000, Cell Signaling Technology, Dancers, MA, USA), ZFP36 (ab204922, 1: 5000, Abcam, Cambridge, UK), IL-10 (ab189392, 1: 1000, Abcam), Foxp3 (ab215206, 1: 1000, Abcam), and TGF-β (ab215715, 1: 1000, Abcam) were allowed to incubate with the membrane overnight at 4°C.

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Control

Journal: Journal of Asthma and Allergy

Article Title: MALAT1 Induces Food Allergy by Promoting Release of IL-6 from Dendritic Cells and Suppressing the Immunomodulatory Function of Tregs

doi: 10.2147/JAA.S341742

Figure Lengend Snippet: MALAT1 aggravates FA by promoting release of IL-6 from DCs and inhibiting Treg immunomodulatory function through downregulation of ZFP36. ( A ) Representative H&E staining images of mouse jejunal tissues to observe inflammation and flow cytometric quantification of eosinophils and monocytes. ( B ) Flow cytometry quantification of mast cells observed by Toluidine blue staining. ( C ) CD4 + CD25 + Foxp3 + cell ratio in mouse splenocytes. ( D ) Immunohistochemical detection of Foxp3 + cell ratio in mouse jejunal tissues. ( E ) ELISA to measure IL-10 and TGF-β expression levels in mouse splenocyte supernatant (in panels ( B – E ), n = 10, * p < 0.05 vs the control group, # p < 0.05 vs the model + oe-NC group). All cellular experiments were repeated 3 times independently.

Article Snippet: Protein separation was done using sodium dodecyl sulfate polyacrylamide gel electrophoresis and the proteins were transferred onto a polyvinylidene fluoride membrane (Millipore, Burlington, MA, USA) by wet transfer method, followed by blocking at ambient temperature with 5% bovine serum albumin for 1 h. Subsequently, diluted primary antibodies against β-actin (4970, 1: 5000, Cell Signaling Technology, Dancers, MA, USA), ZFP36 (ab204922, 1: 5000, Abcam, Cambridge, UK), IL-10 (ab189392, 1: 1000, Abcam), Foxp3 (ab215206, 1: 1000, Abcam), and TGF-β (ab215715, 1: 1000, Abcam) were allowed to incubate with the membrane overnight at 4°C.

Techniques: Staining, Flow Cytometry, Immunohistochemical staining, Enzyme-linked Immunosorbent Assay, Expressing, Control

Journal: Journal of Asthma and Allergy

Article Title: MALAT1 Induces Food Allergy by Promoting Release of IL-6 from Dendritic Cells and Suppressing the Immunomodulatory Function of Tregs

doi: 10.2147/JAA.S341742

Figure Lengend Snippet: Molecular mechanism of MALAT1-mediated ZFP36 affecting the immunomodulatory function of DCs and Tregs in FA.

Article Snippet: Protein separation was done using sodium dodecyl sulfate polyacrylamide gel electrophoresis and the proteins were transferred onto a polyvinylidene fluoride membrane (Millipore, Burlington, MA, USA) by wet transfer method, followed by blocking at ambient temperature with 5% bovine serum albumin for 1 h. Subsequently, diluted primary antibodies against β-actin (4970, 1: 5000, Cell Signaling Technology, Dancers, MA, USA), ZFP36 (ab204922, 1: 5000, Abcam, Cambridge, UK), IL-10 (ab189392, 1: 1000, Abcam), Foxp3 (ab215206, 1: 1000, Abcam), and TGF-β (ab215715, 1: 1000, Abcam) were allowed to incubate with the membrane overnight at 4°C.

Techniques:

Journal: bioRxiv

Article Title: The Hemopurifier removes extracellular vesicles and microRNAs from renal perfusates following controlled oxygenated rewarming of discarded donor kidneys

doi: 10.1101/2024.08.23.609252

Figure Lengend Snippet: (A) NanoFCM nanoparticle flow cytometry analysis demonstrates that the Hemopurifier removes the majority of small EV particles (40-200nm nanovesicles) and (B) of dsDNA loaded EV particles in treated renal perfusates. The data shown represents the mean and standard error of 2 technical replicates measured from each perfusate sample.(* p ≤ 0.05; **p ≤ 0.01)

Article Snippet: Concentration of EV and dsDNA-associated EV nanoparticles was determined through the use of high sensitivity nanoparticle NanoFCM Flow Analyzer U30 (NanoFCM, Nottingham, UK).

Techniques: Flow Cytometry

Journal: ACS Omega

Article Title: Impact of Environmental Pollutant Cadmium on the Establishment of a Cancer Stem Cell Population in Breast and Hepatic Cancer

doi: 10.1021/acsomega.6b00181

Figure Lengend Snippet: Schematic illustration of the monitoring of cadmium-induced CSCs based on the concurrent identification of multiple CSC markers using QD and AOTF-based cellular imaging. MCF-7 and HepG2 cells were treated with cadmium for 72 h, followed by the separation of CSCs using a magnetic sorter. Then, hepatic CSCs were identified based on the expression of QD-conjugated CD133, CD44, and ALDH1, and breast CSCs were identified based on the expression of CD44, CD24, and ALDH1. The AOTF scans the cellular images at a single wavelength emitted from the QD-based nanoprobes without spectral overlap.

Article Snippet: Antibodies specific for CSC markers CD133 (ab-19898; Abcam, USA), CD44 (sc-65265), CD24 (sc-19585), and ALDH1 (sc-374149) (Santa Cruz biotechnology, Santa Cruz, USA) were conjugated to QD705, QD525, QD565, and QD625, respectively, according to the manufacturer’s instructions (Quantum Dot Conjugation Kit; Invitrogen, Carlsbad, CA, USA).

Techniques: Imaging, Expressing

Journal: ACS Omega

Article Title: Impact of Environmental Pollutant Cadmium on the Establishment of a Cancer Stem Cell Population in Breast and Hepatic Cancer

doi: 10.1021/acsomega.6b00181

Figure Lengend Snippet: (a) Breast CSCs obtained from the concurrent monitoring of CD44, CD24, and ALDH1 in the cadmium-treated MCF-7 cell culture. The emission wavelength of QD-conjugated CD44, CD24, and ALDH1 antibodies was measured at 525, 565, and 625 nm, respectively. (b) Quantitative determination of breast CSCs in the MCF-7 cell culture treated at the functional concentration of cadmium. (c) Semiquantitative RT-PCR analysis of CD44, CD24, and ALDH1 expression in MCF-7 cells treated with and without cadmium (1 μM) over the period of 72 h. The values are means ± SD from three independent experiments.

Article Snippet: Antibodies specific for CSC markers CD133 (ab-19898; Abcam, USA), CD44 (sc-65265), CD24 (sc-19585), and ALDH1 (sc-374149) (Santa Cruz biotechnology, Santa Cruz, USA) were conjugated to QD705, QD525, QD565, and QD625, respectively, according to the manufacturer’s instructions (Quantum Dot Conjugation Kit; Invitrogen, Carlsbad, CA, USA).

Techniques: Cell Culture, Functional Assay, Concentration Assay, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: ACS Omega

Article Title: Impact of Environmental Pollutant Cadmium on the Establishment of a Cancer Stem Cell Population in Breast and Hepatic Cancer

doi: 10.1021/acsomega.6b00181

Figure Lengend Snippet: (a–e) FACS analysis for the breast CSC population induced by cadmium in the MCF-7 cell culture. The x -axis represents CD44 emission, and the y -axis represents ALDH1 emission. For the x -axis, the right side indicates CD44 positive; for the y -axis, the upper side indicates ALDH1 positive. (f) Quantitative determination of breast CSCs obtained using flow cytometry.

Article Snippet: Antibodies specific for CSC markers CD133 (ab-19898; Abcam, USA), CD44 (sc-65265), CD24 (sc-19585), and ALDH1 (sc-374149) (Santa Cruz biotechnology, Santa Cruz, USA) were conjugated to QD705, QD525, QD565, and QD625, respectively, according to the manufacturer’s instructions (Quantum Dot Conjugation Kit; Invitrogen, Carlsbad, CA, USA).

Techniques: Cell Culture, Flow Cytometry

Journal: ACS Omega

Article Title: Impact of Environmental Pollutant Cadmium on the Establishment of a Cancer Stem Cell Population in Breast and Hepatic Cancer

doi: 10.1021/acsomega.6b00181

Figure Lengend Snippet: Gene expression of CSC markers in HepG2 cells and MCF-7 cells treated with 25 μM Ras inhibitor and 1 μM cadmium over the period of 72 h. The mRNA expression was analyzed by RT-PCR. (a) mRNA expression of CD44 and ALDH1 in MCF-7 cells. (b) mRNA expression of CD44, CD133, and ALDH1 in HepG2 cells. Values are means ± SD from three independent experiments.

Article Snippet: Antibodies specific for CSC markers CD133 (ab-19898; Abcam, USA), CD44 (sc-65265), CD24 (sc-19585), and ALDH1 (sc-374149) (Santa Cruz biotechnology, Santa Cruz, USA) were conjugated to QD705, QD525, QD565, and QD625, respectively, according to the manufacturer’s instructions (Quantum Dot Conjugation Kit; Invitrogen, Carlsbad, CA, USA).

Techniques: Gene Expression, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: iScience

Article Title: Preoperative immune checkpoint inhibition and cryoablation in early-stage breast cancer

doi: 10.1016/j.isci.2024.108880

Figure Lengend Snippet:

Article Snippet: Briefly, one million PBMCs and TILs were washed with 2 mL FACS buffer (PBS [phosphate-buffered saline] containing bovine 1% serum albumin and 0.05 mM EDTA), resuspended in 50 μL FACS buffer and stained with a fixable Aqua viability dye (Invitrogen) and a cocktail of antibodies to the following surface markers: CD8-Qdot 605 (Invitrogen, 3B5), CD4-Qdot 655 (Invitrogen, S3.5), PD-1-PE (BD, MIH4), LAG-3-FITC (Enzo Life Sciences, 17B4), ICOS-PE-Cy7 (eBioscience, ISA-3), TIM-3-APC (R&D Systems, 344823).

Techniques: Recombinant, Saline, Staining, Extraction, Software, Flow Cytometry