Journal: Materials Today Bio

Article Title: pH-responsive 2D niobium carbide nanosheets for targeted circPUM1 siRNA delivery in ovarian cancer therapy

doi: 10.1016/j.mtbio.2025.102314

Figure Lengend Snippet: Scheme 1. General design of P-Nb 2 C-PEG@si-circPUM1 in ovarian cancer. (A) Schematic illustration of P-Nb 2 C-PEG preparation. The precursor (Nb 2 AlC) underwent HF etching followed by exfoliation with TPAOH, yielding negatively charged monolayer Nb 2 C nanosheets. After mixture with excess PEI, P-Nb 2 C was generated with a positive surface charge. We modified the nanosheets with CHO-PEG-CHO, where the aldehyde groups form Schiff base linkages with amine groups, enabling PEG grafting. (B) P-Nb 2 C-PEG was loaded with circPUM1 siRNA via electrostatic interaction and circulated in blood (pH 7.4). Upon reaching the acidic tumor microenvironment (pH 6.5), the nanosheets underwent pH-responsive charge reversal enabling effective cellular uptake. (C) The internalized circPUM1 siRNA specifically targeted the back-splice junction of circPUM1, disrupted its circular structure and abolished its miRNA-sponging activity. The released miRNAs suppressed the expression of VEGFA and RAB27B, thereby inhibiting RAB27B-mediated exosome secretion and VEGFA-triggered angiogenesis. (Created in BioRender. Guan, X. (2025) https://BioRender.com/x7gxaqp )

Article Snippet: Membranes were subsequently blocked with NcmBlot blocking buffer (NCM Biotech, China) for 10 min. Primary antibody incubation was carried out at 4 °C overnight with specific antibodies against VEGFA (1:1000; Boster, China), RAB27B (1:1000; Boster, China), and β-actin (1:8000; Abbkine, China).

Techniques: Generated, Modification, Activity Assay, Expressing

Journal: Materials Today Bio

Article Title: pH-responsive 2D niobium carbide nanosheets for targeted circPUM1 siRNA delivery in ovarian cancer therapy

doi: 10.1016/j.mtbio.2025.102314

Figure Lengend Snippet: CircPUM1 promotes angiogenesis in ovarian cancer as the form of exosomes. (A) The expression of circPUM1 was positively related with MVD in ovarian cancer tissue. (B) Experimental design schematic (Created in BioRender. Guan, X. (2025) https://BioRender.com/f5br0q0 ): HUVEC were co-cultured with exosomes derived from circPUM1-overexpressing and control OVCAR3 cells. (C) RT-qPCR showed that circPUM1 was highly expressed in HUVEC after co-cultured with exosomal circPUM1. Exosomal circPUM1 enhanced migration ability (D, E), viability (F), invasiveness (G), and tube formation ability (H) of HUVECs. (I) Circular RNA pull-down assays using biotinylated circPUM1 probes confirmed co-enrichment of circPUM1 and miR-607. (J) Dual-luciferase reporter assays showed miR-607 targeting 3′-UTR in both VEGFA and RAB27B . (K) Western blot revealed that overexpression of circPUM1 upregulated VEGFA and RAB27B , whereas miR-607 downregulated these targets in OVCAR3 cells. Exosomal circPUM1 elevated intracellular VEGFA expression in HUVECs. (L) Molecular schematic (Created in BioRender. Guan, X. (2025) https://BioRender.com/5eldsz5 ): In ovarian cancer, the highly expressed circPUM1 sponges miR-607, upregulating expression of VEGFA and RAB27B . This dual regulation establishes a pro-tumorigenic cascade through two distinct pathways: (1) RAB27B -mediated secretion of exosomal circPUM1, and (2) VEGFA -induced activation of angiogenic signaling.

Article Snippet: Membranes were subsequently blocked with NcmBlot blocking buffer (NCM Biotech, China) for 10 min. Primary antibody incubation was carried out at 4 °C overnight with specific antibodies against VEGFA (1:1000; Boster, China), RAB27B (1:1000; Boster, China), and β-actin (1:8000; Abbkine, China).

Techniques: Expressing, Cell Culture, Derivative Assay, Control, Quantitative RT-PCR, Migration, Luciferase, Western Blot, Over Expression, Activation Assay

Journal: Materials Today Bio

Article Title: pH-responsive 2D niobium carbide nanosheets for targeted circPUM1 siRNA delivery in ovarian cancer therapy

doi: 10.1016/j.mtbio.2025.102314

Figure Lengend Snippet: In vitro antitumor performance of P-Nb 2 C@si-circPUM1 in ovarian cancer cells. (A) Confocal microscopy images showed intracellular uptake of Cy3-labeled circPUM1 siRNA. Cell apoptosis assays (B), Transwell assays (C), wound healing assays (E) and EdU proliferation assays (F) showed that P-Nb 2 C-loaded circPUM1 siRNA effectively suppressed ovarian cancer cell phenotypes. (G) RT-qPCR demonstrated a significant downregulation of circPUM1 expression in lipo2000-transfected and P-Nb 2 C-loaded siRNA groups. Immunofluorescence (D) and Western blot (H) validated the downregulation of VEGFA and RAB27B expression in cells treated with P-Nb 2 C@si-circPUM1. (I) CCK8 assays showed inhibition of ovarian cancer cell viability in lipo2000-transfected and P-Nb 2 C-loaded siRNA groups.

Article Snippet: Membranes were subsequently blocked with NcmBlot blocking buffer (NCM Biotech, China) for 10 min. Primary antibody incubation was carried out at 4 °C overnight with specific antibodies against VEGFA (1:1000; Boster, China), RAB27B (1:1000; Boster, China), and β-actin (1:8000; Abbkine, China).

Techniques: In Vitro, Confocal Microscopy, Labeling, Quantitative RT-PCR, Expressing, Transfection, Immunofluorescence, Western Blot, Inhibition

Journal: Materials Today Bio

Article Title: pH-responsive 2D niobium carbide nanosheets for targeted circPUM1 siRNA delivery in ovarian cancer therapy

doi: 10.1016/j.mtbio.2025.102314

Figure Lengend Snippet: In vitro anti-angiogenic performance of P-Nb 2 C@si-circPUM1 in HUVECs. (A) QPCR showed a reduction of circPUM1 levels in exosomes derived from P-Nb 2 C@si-circPUM1-treated ovarian cancer cells. (B) Intracellular circPUM1 level in HUVEC treated with Exo-P-Nb 2 C@si-circPUM1 were markedly lower than those in Exo-control and Exo-P-Nb 2 C@si-scramble groups. CCK-8 and EdU assays demonstrated that Exo-P-Nb 2 C@si-circPUM1 suppressed cell viability (C) and DNA replication capacity (D, E) in HUVECs. Wound healing, Transwell assay and tube formation assay indicated that Exo-P-Nb 2 C@si-circPUM1 treatment significantly attenuated HUVEC migration (F), invasion (G), and angiogenic potential (I) compared to control groups. Immunofluorescence (H, J) and Western blot (K) confirmed downregulation of VEGFA in Exo-P-Nb 2 C@si-circPUM1 treated HUVECs.

Article Snippet: Membranes were subsequently blocked with NcmBlot blocking buffer (NCM Biotech, China) for 10 min. Primary antibody incubation was carried out at 4 °C overnight with specific antibodies against VEGFA (1:1000; Boster, China), RAB27B (1:1000; Boster, China), and β-actin (1:8000; Abbkine, China).

Techniques: In Vitro, Derivative Assay, Control, CCK-8 Assay, Transwell Assay, Tube Formation Assay, Migration, Immunofluorescence, Western Blot

Journal: Materials Today Bio

Article Title: pH-responsive 2D niobium carbide nanosheets for targeted circPUM1 siRNA delivery in ovarian cancer therapy

doi: 10.1016/j.mtbio.2025.102314

Figure Lengend Snippet: Molecular mechanism validation of P-Nb 2 C-PEG@si-circPUM1 anti-angiogenic performance in vivo . (A) H&E staining showed increased necrotic loci within tumors of P-Nb 2 C-PEG@si-circPUM1 group. (B, D) IHC staining of Ki-67 demonstrated a clear reduction in Ki-67-positive cells in P-Nb 2 C-PEG@si-circPUM1 group. (C, F) IHC staining of CD31 indicated a marked decrease in MVD within tumors of P-Nb 2 C-PEG@si-circPUM1 group. (H) QPCR showed a significant downregulation of circPUM1 expression in the P-Nb 2 C-PEG@si-circPUM1 treated group. IHC (E, G) and Western blot (I, J) demonstrated significant down-regulation of RAB27B and VEGFA .

Article Snippet: Membranes were subsequently blocked with NcmBlot blocking buffer (NCM Biotech, China) for 10 min. Primary antibody incubation was carried out at 4 °C overnight with specific antibodies against VEGFA (1:1000; Boster, China), RAB27B (1:1000; Boster, China), and β-actin (1:8000; Abbkine, China).

Techniques: Biomarker Discovery, In Vivo, Staining, Immunohistochemistry, Expressing, Western Blot

Journal: bioRxiv

Article Title: The genomic and transcriptional landscape of the spinal cord H3K27-altered diffuse midline glioma suggests the potential therapeutic strategy

doi: 10.1101/2025.05.25.655886

Figure Lengend Snippet: (A) Schematic of workflow for generating patient-derived spinal cord H3K27-altered DMG cell cultures. (B) Micrographs of immunofluorescence in spinal cord H3K27-altered DMG neurospheres stained for the indicated marker of each meta-module (red) and DAPI (blue). (C) Response of DMG neurospheres to combined panobinostat (Pan) and tazemetostat (Taz) or DMSO control after incubation for 72 hours. (D) Relative expression of feature genes of each meta-module in DMG neurospheres after 24, 48, and 72 hours of incubation with DMSO control or combined panobinostat (Pan) and tazemetostat (Taz). Results are shown in columns of duplicates. (E) Western blots of VEGFA, S100A11, GADD45A, SHMT2, and GAPDH in DMG neurospheres after incubation with combined epigenetic agents or DMSO for 48 hours are shown in the top left panel. The respective protein levels in epigenetic-agent-treated cells relative to DMSO control are also shown. **, p<0.01, ***, p<0.001, Student’s t test.

Article Snippet: After blockade with 5% skimmed milk dissolved in TBST composed of 1×TBS (Lablead) and 0.1% TWEEN 20 (VWR), protein blots were incubated with primary antibodies against VEGFA (Proteintech), S100A11 (Proteintech), GADD45A (Proteintech), and SHMT2 (Proteintech), followed by horseradish peroxidase-conjugated goat anti-rabbit (EASYBIO) or anti-mouse (EASYBIO) secondary antibodies.

Techniques: Derivative Assay, Immunofluorescence, Staining, Marker, Control, Incubation, Expressing, Western Blot

Journal: Renal Failure

Article Title: miR-29a-3p/ Vegfa axis modulates high phosphate-induced vascular smooth muscle cell calcification

doi: 10.1080/0886022x.2025.2489712

Figure Lengend Snippet: Figure 1. Identification of Vegfa as a direct target of miR-29a-3p in VSMCs using bioinformatics and dual-luciferase assay. (A) Venn diagram depicting the overlap of miR-29a-3p target genes predicted by four miRNA target prediction tools: miRTarBase, miRWalk, TargetScan, and miRDB. The box highlights the seven genes identified as common targets across all tools. (B) Sequence alignment of miR-29a-3p with the 3′-UTR region of wild-type and mutated Vegfa mRNA. The predicted binding site and seed region are marked in red. (C) Dual-luciferase reporter assay results for VSMCs co-transfected with a control vector, a vector containing the wild type (WT) Vegfa 3′-UTR sequence, or the MUT Vegfa sequence, along with miR-29a-3p mimics or a negative control miRNA (NT). Relative luciferase activity was measured, and data are presented as mean ± SD from three independent experiments.

Article Snippet: VSMCs recovered by centrifugation were homogenized and separated using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (China National Pharmaceutical Group Corporation Chemical reagent Co., Ltd., China; Cat. #30166428). the proteins were then transferred to polyvinylidene fluoride membranes (Millipore Co., uSa; Cat. #ISEQ00010). the blots were stained with rabbit polyclonal antibodies against VEGFa (1:10,000; Proteintech, uSa; Cat. #19003-1-aP), and mouse monoclonal antibodies against β-actin (1:10,000, Servicebio, China; Cat. #Gb15001), followed by anti-rabbit IgG (1:10,000, SIMubIotECH, China; Cat. #S2001), and anti-mouse IgG horseradish peroxidase conjugates (1:10,000, abclonal, China; Cat. #aS003).

Techniques: Luciferase, Sequencing, Binding Assay, Reporter Assay, Transfection, Control, Plasmid Preparation, Negative Control, Activity Assay

Journal: Renal Failure

Article Title: miR-29a-3p/ Vegfa axis modulates high phosphate-induced vascular smooth muscle cell calcification

doi: 10.1080/0886022x.2025.2489712

Figure Lengend Snippet: Figure 2. miR-29a-3p overexpression suppresses Vegfa expression in VSMCs. (A) Representative Western blot analysis of VEGFA protein levels in VSMCs under standard culture conditions (control), high phosphate conditions (Pi), and high phosphate conditions following miR-29a-3p overexpression (Pi + miR-29a-3p). β-actin served as a loading control. Quantitative analysis of VEGFA protein levels is shown as relative fold change normalized to β-actin. (B) Vegfa mRNA levels quantified using RT-qPCR in the same experimental groups. GAPDH was used as an internal control for normalization. Data represent the mean ± SD of five independent biological replicates.

Article Snippet: VSMCs recovered by centrifugation were homogenized and separated using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (China National Pharmaceutical Group Corporation Chemical reagent Co., Ltd., China; Cat. #30166428). the proteins were then transferred to polyvinylidene fluoride membranes (Millipore Co., uSa; Cat. #ISEQ00010). the blots were stained with rabbit polyclonal antibodies against VEGFa (1:10,000; Proteintech, uSa; Cat. #19003-1-aP), and mouse monoclonal antibodies against β-actin (1:10,000, Servicebio, China; Cat. #Gb15001), followed by anti-rabbit IgG (1:10,000, SIMubIotECH, China; Cat. #S2001), and anti-mouse IgG horseradish peroxidase conjugates (1:10,000, abclonal, China; Cat. #aS003).

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

Journal: Renal Failure

Article Title: miR-29a-3p/ Vegfa axis modulates high phosphate-induced vascular smooth muscle cell calcification

doi: 10.1080/0886022x.2025.2489712

Figure Lengend Snippet: Figure 3. Vegfa knockdown and miR-29a-3p overexpression mitigate high phosphate-induced VSMCs calcification. (A–C) Analysis of Vegfa knockdown effects in VSMCs cultured in normal media (control) or high phosphate conditions (Pi). (A) Vegfa mRNA levels were quantified via RT-qPCR. (B) ARS with corresponding spectrophotometric quantification to assess cellular mineralization. (C) Quantification of intracellular Ca2+ content. Cells were treated with small interfering RNA targeting Vegfa (siVegfa) or a NT control. (D–F) Investigation of the role of miR-29a-3p in VSMCs cultured under high phosphate conditions. (D) Vegfa mRNA expression measured using RT-qPCR, (E) mineralization assessed using ARS and spectrophotometric quantification, and (F) intracellular Ca2+ levels were quantified. Cells were transfected with either an empty vector (VE) or a Vegfa-overexpressing construct (OE), combined with a NT miRNA mimic or a miR-29a-3p mimic. All experiments were performed in five replicates, and error bars represent the SD. GAPDH was used as an internal control in RT-qPCR experiments. Scale bar = 100 µm.

Article Snippet: VSMCs recovered by centrifugation were homogenized and separated using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (China National Pharmaceutical Group Corporation Chemical reagent Co., Ltd., China; Cat. #30166428). the proteins were then transferred to polyvinylidene fluoride membranes (Millipore Co., uSa; Cat. #ISEQ00010). the blots were stained with rabbit polyclonal antibodies against VEGFa (1:10,000; Proteintech, uSa; Cat. #19003-1-aP), and mouse monoclonal antibodies against β-actin (1:10,000, Servicebio, China; Cat. #Gb15001), followed by anti-rabbit IgG (1:10,000, SIMubIotECH, China; Cat. #S2001), and anti-mouse IgG horseradish peroxidase conjugates (1:10,000, abclonal, China; Cat. #aS003).

Techniques: Knockdown, Over Expression, Cell Culture, Control, Quantitative RT-PCR, Small Interfering RNA, Expressing, Transfection, Plasmid Preparation, Construct