Journal: Frontiers in Immunology

Article Title: Protective effects of conditioned media of immortalized stem cells from human exfoliated deciduous teeth on pressure ulcer formation

doi: 10.3389/fimmu.2022.1010700

Figure Lengend Snippet: Inhibitory effects of SHED-CM on PU formation are highly dependent on VEGF and HGF. (A) Cutaneous I/R injury was performed on the dorsal skin of mice using two magnetic plates to induce PU formation. After cutaneous I/R injury was induced, 100 µl SHED-CM, VEGF-depleted SHED-CM (B) , HGF-depleted SHED-CM (D) , or each control antibody-treated SHED-CM (C, E) was injected into the dermis at two sites around the wound at the indicated time points. The size of the wound areas in each photograph was evaluated using FIJI, and the relative wound areas at each time point to the maximum area in control mice on day 3~5 as 100% were calculated. Data are shown as the mean ± SEM (n = 4~5) and are representative of two to three independent experiments. P values were determined by the unpaired two-tailed Student’s t-test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Briefly, SHED-CM was incubated with antibody against VEGF (clone R012; Sino Biological), HGF (clone 24612; R&D Systems), IGFBP-4 (clone 82314; R&D Systems) or respective control IgG conjugated to protein G-Sepharose (GE Healthcare) overnight at 4°C.

Techniques: Injection, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Protective effects of conditioned media of immortalized stem cells from human exfoliated deciduous teeth on pressure ulcer formation

doi: 10.3389/fimmu.2022.1010700

Figure Lengend Snippet: SHED-CM promotes angiogenesis through VEGF and HGF. (A, C) Mouse dorsal epidermal tissues of ulcers were removed on day 4 and immunohistochemically analyzed for expression of the endothelial cell marker CD31 and pericyte marker NG2 counterstaining with Mayer’s hematoxylin. Representative photographs for CD31 (A) or NG2 (C) are shown. Positive areas for CD31 or NG2 were calculated using FIJI (B, D) . Data are shown as the mean ± SEM (n = 5~6) and are representative of two independent experiments. (E) Angiogenic activities of SHED-CM, control medium, or VEGF- or HGF-depleted SHED-CM with respective antibodies, control antibodies-treated SHED-CM were determined using a tube formation assay with HUVECs. Representative photographs of the tube formation are shown. (F) Total tube area and branch formation rate were quantified using FIJI. Data are shown as the mean ± SEM in triplicate and are representative of five independent experiments. P values were determined by one-way analysis of variance with the Tukey multiple comparisons test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Briefly, SHED-CM was incubated with antibody against VEGF (clone R012; Sino Biological), HGF (clone 24612; R&D Systems), IGFBP-4 (clone 82314; R&D Systems) or respective control IgG conjugated to protein G-Sepharose (GE Healthcare) overnight at 4°C.

Techniques: Expressing, Marker, Tube Formation Assay

Journal: Frontiers in Immunology

Article Title: Protective effects of conditioned media of immortalized stem cells from human exfoliated deciduous teeth on pressure ulcer formation

doi: 10.3389/fimmu.2022.1010700

Figure Lengend Snippet: SHED-CM suppresses ROS generation through VEGF and HGF. (A) Mouse dorsal epidermal tissues of ulcers were removed on day 4 and immunohistochemically analyzed for expression of the oxidative stress marker 8-OHdG counterstaining with Mayer’s hematoxylin. Representative photographs for 8-OHdG are shown. Subcutaneous adipose tissue, sebaceous gland, hair bulb, dermis and epidermis were positively stained. (B) Positive areas for 8-OHdG were calculated using FIJI. Data are shown as the mean ± SEM (n = 5~6) and are representative of two independent experiments. (C) The effects of SHED-CM on ROS generation induced by hypoxia/reoxygenation in vitro was examined. NIH3T3 cells were incubated under hypoxic condition (1% O 2 , 5% CO 2 , and 94% N 2 ) for 48 h in the medium containing 1% FBS, and reoxygenation was then performed under normal ambient O 2 conditions (21%) in the presence of the SHED-CM, control medium, or VEGF- or HGF-depleted SHED-CM with respective antibodies, control antibodies-treated SHED-CM. After 6 h, cells were counterstained with Hoechst and analyzed for intracellular ROS production. Representative photographs of the ROS generation are shown. (D) The fluorescence intensity of DCF was quantified using FIJI and calculated as arbitrary unit per cell. Data are shown as the mean ± SEM in triplicate and are representative of two independent experiments. P values were determined by the one-way analysis of variance with the Tukey multiple comparisons test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Briefly, SHED-CM was incubated with antibody against VEGF (clone R012; Sino Biological), HGF (clone 24612; R&D Systems), IGFBP-4 (clone 82314; R&D Systems) or respective control IgG conjugated to protein G-Sepharose (GE Healthcare) overnight at 4°C.

Techniques: Expressing, Marker, Staining, In Vitro, Incubation, Fluorescence

Journal: Journal of Endocrinology

Article Title: The effects of recombinant human GH on promoting tumor growth depend on the expression of GH receptor in vivo

doi: 10.1530/joe-11-0100

Figure Lengend Snippet: Figure 3 The VEGF expression in tumor tissues. (A–C) VEGF staining in SGC-7901 observed with high microscope (200!original magnifications). (D–F) VEGF staining in MKN-45 observed with high microscope (200! original magnifications). A and D, control groups; B and E, low-dose rhGH groups; and C and F, high-dose rhGH groups. Three animals per group were used for immunohistochemistry assay. Full colour version of this figure available via http://dx.doi.org/10.1530/ JOE-11-0100.

Article Snippet: BALB-c/nunu male nude mice (4–5 weeks old, 14–16 g) were purchased from the Shanghai Laboratory Animal Center of the Chinese Academy of Sciences. rhGH was obtained from Serono Pharm Co., (Geneva, Switzerland); RPMI medium 1640 was purchased from Gibco Co.; rabbit anti-human GHR polyclonal antibody, rabbit anti-human VEGF monoclonal antibody, and HRPlabeled goat anti-rabbit secondary antibody were purchased from Boster Biological Company of China (Wuhan, China); signal transducers and activators of transcription 3 (STAT3), phosphorylated STAT3 (pSTAT3), hypoxia-inducible factor 1, alpha subunit (HIF-a), matrix metalloproteinase 2 (MMP-2), and other antibodies were purchased from Santa Cruz Co., (Santa Cruz, CA, USA).

Techniques: Expressing, Staining, Microscopy, Control, Immunohistochemistry

Journal: Journal of Endocrinology

Article Title: The effects of recombinant human GH on promoting tumor growth depend on the expression of GH receptor in vivo

doi: 10.1530/joe-11-0100

Figure Lengend Snippet: Figure 4 The mRNA expressions of angiogenesis-related factors in tumor tissues. (A) RT-PCR analysis of Ghr, Jak-2, Stat3, Vegf, Hif-1a, Fgf, and Mmp-2. Sc, control group of SGC-7901; Sl, low-rhGH treatment group of SGC-7901; Sh, high-rhGH treatment group of SGC-7901; Mc, control group of MKN-45; Ml, low-rhGH treatment group of MKN-45; Mh, high-rhGH treatment group of MKN-45. (B) Ratio of gray scale in SGC-7901 groups (*P!0.05 vs control and #P!0.05 vs low-dose rhGH group). (C) Ratio of gray scale in MKN-45 groups. Four animals per group were used for RT-PCR.

Article Snippet: BALB-c/nunu male nude mice (4–5 weeks old, 14–16 g) were purchased from the Shanghai Laboratory Animal Center of the Chinese Academy of Sciences. rhGH was obtained from Serono Pharm Co., (Geneva, Switzerland); RPMI medium 1640 was purchased from Gibco Co.; rabbit anti-human GHR polyclonal antibody, rabbit anti-human VEGF monoclonal antibody, and HRPlabeled goat anti-rabbit secondary antibody were purchased from Boster Biological Company of China (Wuhan, China); signal transducers and activators of transcription 3 (STAT3), phosphorylated STAT3 (pSTAT3), hypoxia-inducible factor 1, alpha subunit (HIF-a), matrix metalloproteinase 2 (MMP-2), and other antibodies were purchased from Santa Cruz Co., (Santa Cruz, CA, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Control

Journal: Journal of Endocrinology

Article Title: The effects of recombinant human GH on promoting tumor growth depend on the expression of GH receptor in vivo

doi: 10.1530/joe-11-0100

Figure Lengend Snippet: Figure 5 The protein expressions of angiogenesis-related factors in tumor tissues. (A) Western blot analysis of STAT3, pSTAT3, VEGF, HIF-1a, and MMP-2. Sc, control group of SGC-7901; Sl, low-rhGH treatment group of SGC-7901; Sh, high-rhGH treatment group of SGC-7901; Mc, control group of MKN-45; Ml, low-rhGH treatment group of MKN-45; Mh, high-rhGH treatment group of MKN-45. (B) Ratio of gray scale in SGC-7901 groups (*P!0.05 vs control and #P!0.05 vs low-dose rhGH group). (C) Ratio of gray scale in MKN-45 groups. Four animals per group were used for western blotting.

Article Snippet: BALB-c/nunu male nude mice (4–5 weeks old, 14–16 g) were purchased from the Shanghai Laboratory Animal Center of the Chinese Academy of Sciences. rhGH was obtained from Serono Pharm Co., (Geneva, Switzerland); RPMI medium 1640 was purchased from Gibco Co.; rabbit anti-human GHR polyclonal antibody, rabbit anti-human VEGF monoclonal antibody, and HRPlabeled goat anti-rabbit secondary antibody were purchased from Boster Biological Company of China (Wuhan, China); signal transducers and activators of transcription 3 (STAT3), phosphorylated STAT3 (pSTAT3), hypoxia-inducible factor 1, alpha subunit (HIF-a), matrix metalloproteinase 2 (MMP-2), and other antibodies were purchased from Santa Cruz Co., (Santa Cruz, CA, USA).

Techniques: Western Blot, Control

Journal: Scientific Reports

Article Title: HIF-1α and HIF-2α induced angiogenesis in gastrointestinal vascular malformation and reversed by thalidomide

doi: 10.1038/srep27280

Figure Lengend Snippet: ( A ) The expression of HIF-1α and HIF-2α in gastrointestinal vascular malformations and normal vessels. Red arrow: strongly positive; Black arrow: weakly positive; Blue arrow: negative. ( B ) Percentages of positive and negative vessels in GIVM and normal tissues. ** P < 0.01.

Article Snippet: Proteins were transferred to a nitrocellulose membrane (Millipore, Bedford, MA, USA), which was blocked using 5% milk for 2 h, and incubated with rabbit-anti-HIF-2α, VEGF, NOTCH1, DLL4, Ang2, and β-actin antibodies (all rabbit polyclonal antibodies from KangChen Bio-tech, Shanghai, China) at 4 °C overnight.

Techniques: Expressing

Journal: Scientific Reports

Article Title: HIF-1α and HIF-2α induced angiogenesis in gastrointestinal vascular malformation and reversed by thalidomide

doi: 10.1038/srep27280

Figure Lengend Snippet: ( A ) Western blot determinations of HIF-1α and HIF-2α expression at different time points of hypoxia. * P < 0.05, ** P < 0.01 vs. 0 hour. ( B ) The effect of HIF-1α and HIF-2α overexpression on the expression of VEGF, Notch1, DLL4, and Ang2. Western blot and RT-PCR demonstrated that HIF-1α and HIF-2α overexpression increased the expression of VEGF, Notch1, DLL4, and Ang2 protein and mRNA. * P < 0.05, ** P < 0.01 vs. control. ( C ) Influence of HIF-1α and HIF-2α overexpression on angiogenesis 6 and 24 h after transfection of Lenti-HIF-1α and Lenti-HIF2α. Tube formation was enhanced 6 and 24 h after transfection. ** P < 0.01 vs. control. ( D ) Fluorescence microscope observations of subintestinal vein sprouting in normal and HIF-2α-overexpressing zebrafish. *Indicates subintestinal vascular sprouts. HIF-2α overexpression significantly increased the number of subintestinal vascular sprouts. ** P < 0.01 vs. control plasmid. ( E ) Dual luciferase reporter gene assay demonstrated that HIF-2α enhanced VEGF promoter activity. ** P < 0.01 vs. control plasmid.

Article Snippet: Proteins were transferred to a nitrocellulose membrane (Millipore, Bedford, MA, USA), which was blocked using 5% milk for 2 h, and incubated with rabbit-anti-HIF-2α, VEGF, NOTCH1, DLL4, Ang2, and β-actin antibodies (all rabbit polyclonal antibodies from KangChen Bio-tech, Shanghai, China) at 4 °C overnight.

Techniques: Western Blot, Expressing, Over Expression, Reverse Transcription Polymerase Chain Reaction, Transfection, Fluorescence, Microscopy, Plasmid Preparation, Luciferase, Reporter Gene Assay, Activity Assay

Journal: Scientific Reports

Article Title: HIF-1α and HIF-2α induced angiogenesis in gastrointestinal vascular malformation and reversed by thalidomide

doi: 10.1038/srep27280

Figure Lengend Snippet: ( A ) Immunofluorescence indicated that HIF-1α and HIF-2α expression was down-regulated by thalidomide at different concentrations. ( B ) Western blots demonstrated that the expression of HIF-1α and HIF-2α decreased with 100 and 200 μg/ml of thalidomide. * P < 0.05, ** P < 0.01. ( C ) Western blots demonstrated that thalidomide at 200 μg/ml inhibited the expression of HIF-1α and HIF-2α in HUVECs after hypoxic treatment for 24, 36, and 48 h. * P < 0.05, ** P < 0.01. ( D ) Fluorescence microscope observations of the effect of thalidomide at different concentrations on vascular development in zebrafish with HIF-2α overexpression. ** P < 0.01 vs. HIF-2α.

Article Snippet: Proteins were transferred to a nitrocellulose membrane (Millipore, Bedford, MA, USA), which was blocked using 5% milk for 2 h, and incubated with rabbit-anti-HIF-2α, VEGF, NOTCH1, DLL4, Ang2, and β-actin antibodies (all rabbit polyclonal antibodies from KangChen Bio-tech, Shanghai, China) at 4 °C overnight.

Techniques: Immunofluorescence, Expressing, Western Blot, Fluorescence, Microscopy, Over Expression

Journal: Biomedicines

Article Title: Differential Expression of CD3, TNF-α, and VEGF Induced by Olanzapine on the Spleen of Adult Male Albino Rats and the Possible Protective Role of Vitamin C

doi: 10.3390/biomedicines7020039

Figure Lengend Snippet: Photomicrographs of immunohistochemically-stained sections of the spleen with anti- vascular endothelial growth factor (VEGF) from different studied groups. ( A ) Control group, ( B ) vitamin C group showing positive brown immunoreactivity (arrows) within the red pulp, with negatively stained white pulp. ( C ) Olanzapine-treated group showing a marked increase in immunostaining within the red pulp (arrow) and negatively stained white pulp. Note, strong brown cytoplasmic staining within endothelial cells lining the venous sinuses. ( D ) Olanzapine and vitamin C group showing moderate immunostaining within the red pulp in the endothelial cells of venous sinuses (arrow) but no immunoreactivity within the white pulp. VEGF immunostaining, ×100.

Article Snippet: Sections of all studied groups were stained with rabbit monoclonal antibody (SP7) (abcam, 16669, Cambridge, UK) against CD3 at dilution 1:100, rabbit polyclonal antibody against TNF-α (abcam, ab6671, Cambridge, UK) at 1:200, and the rabbit polyclonal antibody against VEGF at 1:500 (Gene Tex, USA).

Techniques: Staining, Control, Immunostaining

Journal: Biomedicines

Article Title: Differential Expression of CD3, TNF-α, and VEGF Induced by Olanzapine on the Spleen of Adult Male Albino Rats and the Possible Protective Role of Vitamin C

doi: 10.3390/biomedicines7020039

Figure Lengend Snippet: Area percentage of VEGF of spleens in group (III) (Olanzapine-treated group) was significantly increased compared to group (I) (control group). The Olanzapine and vitamin C group (IV) was significantly decreased as compared to group (III). The differences in area percentage of VEGF between group (I) and group (II) were not significant ( p > 0.05). Results are expressed as mean ± SD and the level of significance was set for p -values less than 0.05. **: significant increase, group (III) versus group (I) ( p < 0.001). *: significant decrease, group (IV) versus group (III) ( p < 0.001).

Article Snippet: Sections of all studied groups were stained with rabbit monoclonal antibody (SP7) (abcam, 16669, Cambridge, UK) against CD3 at dilution 1:100, rabbit polyclonal antibody against TNF-α (abcam, ab6671, Cambridge, UK) at 1:200, and the rabbit polyclonal antibody against VEGF at 1:500 (Gene Tex, USA).

Techniques: Control