Journal: Techniques in Coloproctology

Article Title: Comparison of the modeling effects between two hemorrhoid models

doi: 10.1007/s10151-026-03303-x

Figure Lengend Snippet: Histology and IL-2 expression in rat hemorrhoidal tissue. A Microscopic appearance of the hemorrhoid areas of the rats in each group (HE, ×50; a model group, b control group). B Expression of IL-2 in anorectal tissue in each group detected by immunohistochemical staining. (IHC, ×50; a model group, b control group)

Article Snippet: The study’s experimental pharmaceuticals and reagents included croton oil (GlpBio 1), olive oil (Sinopharm Chemical Reagent Co., Ltd., 20230807); pyridine (Xilong Scientific Co., Ltd., 240507D1); ether (Tianjin Kemiou Chemical Reagent Co., Ltd., 20230919); 2% isoflurane (RWD Life Science Co., Ltd., 20240902); 4% paraformaldehyde (biosharp BL539A); hematoxylin eosin high definition constant staining kit (Servicebio G1076); bovine serum albumin BSA (Servicebio GC305010 ); normal rabbit serum (concentrated type) (Servicebio G1209); histochemical kit DAB chromogenic reagent (Servicebio G1212); immunohistochemical tumor necrosis factor alpha (TNFα) primary antibody (Servicebio AF7014); immunohistochemical interleukin (IL)-2 primary antibody (Servicebio AF5105); immunohistochemical IL-6 primary antibody (Servicebio DF6087); RIPA lysate (Servicebio G2002-100ML); WB TNFα primary antibody (Servicebio GB12188-100); WB IL-2 primary antibody (Servicebio GB11114-100); WB IL-6 primary antibody (Servicebio GB11117-100); HRP goat anti-rabbit secondary antibody (Servicebio GB23303); HRP goat anti-mouse secondary antibody (Servicebio GB23301); prestained protein marker VII (Servicebio G2087-250UL); RNA extraction solution (Servicebio G3013); isopropanol (Sinopharm Chemical Reagent Co., Ltd., 80109218); absolute ethanol (Sinopharm Chemical Reagent Co., Ltd., 10009218); RNA lysis solution (Servicebio G3029); SweScript All-in-One RT SuperMix for qPCR (Servicebio G3337); 2 × universal blue SYBR Green qPCR Master Mix (Servicebio G3326).

Techniques: Expressing, Control, Immunohistochemical staining, Staining

Journal: Techniques in Coloproctology

Article Title: Comparison of the modeling effects between two hemorrhoid models

doi: 10.1007/s10151-026-03303-x

Figure Lengend Snippet: Immunohistochemical overview of anorectal cytokine expression. Expression of A IL-2, B IL-6, C , and D TNFα in anorectal tissue in each group detected by immunohistochemical staining (IHC, ×50; a model group, b control group)

Article Snippet: The study’s experimental pharmaceuticals and reagents included croton oil (GlpBio 1), olive oil (Sinopharm Chemical Reagent Co., Ltd., 20230807); pyridine (Xilong Scientific Co., Ltd., 240507D1); ether (Tianjin Kemiou Chemical Reagent Co., Ltd., 20230919); 2% isoflurane (RWD Life Science Co., Ltd., 20240902); 4% paraformaldehyde (biosharp BL539A); hematoxylin eosin high definition constant staining kit (Servicebio G1076); bovine serum albumin BSA (Servicebio GC305010 ); normal rabbit serum (concentrated type) (Servicebio G1209); histochemical kit DAB chromogenic reagent (Servicebio G1212); immunohistochemical tumor necrosis factor alpha (TNFα) primary antibody (Servicebio AF7014); immunohistochemical interleukin (IL)-2 primary antibody (Servicebio AF5105); immunohistochemical IL-6 primary antibody (Servicebio DF6087); RIPA lysate (Servicebio G2002-100ML); WB TNFα primary antibody (Servicebio GB12188-100); WB IL-2 primary antibody (Servicebio GB11114-100); WB IL-6 primary antibody (Servicebio GB11117-100); HRP goat anti-rabbit secondary antibody (Servicebio GB23303); HRP goat anti-mouse secondary antibody (Servicebio GB23301); prestained protein marker VII (Servicebio G2087-250UL); RNA extraction solution (Servicebio G3013); isopropanol (Sinopharm Chemical Reagent Co., Ltd., 80109218); absolute ethanol (Sinopharm Chemical Reagent Co., Ltd., 10009218); RNA lysis solution (Servicebio G3029); SweScript All-in-One RT SuperMix for qPCR (Servicebio G3337); 2 × universal blue SYBR Green qPCR Master Mix (Servicebio G3326).

Techniques: Immunohistochemical staining, Expressing, Staining, Control

Journal: Techniques in Coloproctology

Article Title: Comparison of the modeling effects between two hemorrhoid models

doi: 10.1007/s10151-026-03303-x

Figure Lengend Snippet: Relative mRNA expression of pro-inflammatory cytokines A IL-2, B IL-6, and C TNFα in anorectal tissue in each group. * P < 0.05 vs control group

Article Snippet: The study’s experimental pharmaceuticals and reagents included croton oil (GlpBio 1), olive oil (Sinopharm Chemical Reagent Co., Ltd., 20230807); pyridine (Xilong Scientific Co., Ltd., 240507D1); ether (Tianjin Kemiou Chemical Reagent Co., Ltd., 20230919); 2% isoflurane (RWD Life Science Co., Ltd., 20240902); 4% paraformaldehyde (biosharp BL539A); hematoxylin eosin high definition constant staining kit (Servicebio G1076); bovine serum albumin BSA (Servicebio GC305010 ); normal rabbit serum (concentrated type) (Servicebio G1209); histochemical kit DAB chromogenic reagent (Servicebio G1212); immunohistochemical tumor necrosis factor alpha (TNFα) primary antibody (Servicebio AF7014); immunohistochemical interleukin (IL)-2 primary antibody (Servicebio AF5105); immunohistochemical IL-6 primary antibody (Servicebio DF6087); RIPA lysate (Servicebio G2002-100ML); WB TNFα primary antibody (Servicebio GB12188-100); WB IL-2 primary antibody (Servicebio GB11114-100); WB IL-6 primary antibody (Servicebio GB11117-100); HRP goat anti-rabbit secondary antibody (Servicebio GB23303); HRP goat anti-mouse secondary antibody (Servicebio GB23301); prestained protein marker VII (Servicebio G2087-250UL); RNA extraction solution (Servicebio G3013); isopropanol (Sinopharm Chemical Reagent Co., Ltd., 80109218); absolute ethanol (Sinopharm Chemical Reagent Co., Ltd., 10009218); RNA lysis solution (Servicebio G3029); SweScript All-in-One RT SuperMix for qPCR (Servicebio G3337); 2 × universal blue SYBR Green qPCR Master Mix (Servicebio G3326).

Techniques: Expressing, Control

Journal: Techniques in Coloproctology

Article Title: Comparison of the modeling effects between two hemorrhoid models

doi: 10.1007/s10151-026-03303-x

Figure Lengend Snippet: Western blotting for detecting protein expression of IL-2, IL-6, and TNFα

Article Snippet: The study’s experimental pharmaceuticals and reagents included croton oil (GlpBio 1), olive oil (Sinopharm Chemical Reagent Co., Ltd., 20230807); pyridine (Xilong Scientific Co., Ltd., 240507D1); ether (Tianjin Kemiou Chemical Reagent Co., Ltd., 20230919); 2% isoflurane (RWD Life Science Co., Ltd., 20240902); 4% paraformaldehyde (biosharp BL539A); hematoxylin eosin high definition constant staining kit (Servicebio G1076); bovine serum albumin BSA (Servicebio GC305010 ); normal rabbit serum (concentrated type) (Servicebio G1209); histochemical kit DAB chromogenic reagent (Servicebio G1212); immunohistochemical tumor necrosis factor alpha (TNFα) primary antibody (Servicebio AF7014); immunohistochemical interleukin (IL)-2 primary antibody (Servicebio AF5105); immunohistochemical IL-6 primary antibody (Servicebio DF6087); RIPA lysate (Servicebio G2002-100ML); WB TNFα primary antibody (Servicebio GB12188-100); WB IL-2 primary antibody (Servicebio GB11114-100); WB IL-6 primary antibody (Servicebio GB11117-100); HRP goat anti-rabbit secondary antibody (Servicebio GB23303); HRP goat anti-mouse secondary antibody (Servicebio GB23301); prestained protein marker VII (Servicebio G2087-250UL); RNA extraction solution (Servicebio G3013); isopropanol (Sinopharm Chemical Reagent Co., Ltd., 80109218); absolute ethanol (Sinopharm Chemical Reagent Co., Ltd., 10009218); RNA lysis solution (Servicebio G3029); SweScript All-in-One RT SuperMix for qPCR (Servicebio G3337); 2 × universal blue SYBR Green qPCR Master Mix (Servicebio G3326).

Techniques: Western Blot, Expressing

Journal: Techniques in Coloproctology

Article Title: Comparison of the modeling effects between two hemorrhoid models

doi: 10.1007/s10151-026-03303-x

Figure Lengend Snippet: IL-2, IL-6, and TNFα protein levels in anorectal tissue are significantly elevated in the model group compared to the control group (* P < 0.05). A IL-2, B IL-6, and C TNFα protein expression in anorectal tissue. * P < 0.05 vs control group

Article Snippet: The study’s experimental pharmaceuticals and reagents included croton oil (GlpBio 1), olive oil (Sinopharm Chemical Reagent Co., Ltd., 20230807); pyridine (Xilong Scientific Co., Ltd., 240507D1); ether (Tianjin Kemiou Chemical Reagent Co., Ltd., 20230919); 2% isoflurane (RWD Life Science Co., Ltd., 20240902); 4% paraformaldehyde (biosharp BL539A); hematoxylin eosin high definition constant staining kit (Servicebio G1076); bovine serum albumin BSA (Servicebio GC305010 ); normal rabbit serum (concentrated type) (Servicebio G1209); histochemical kit DAB chromogenic reagent (Servicebio G1212); immunohistochemical tumor necrosis factor alpha (TNFα) primary antibody (Servicebio AF7014); immunohistochemical interleukin (IL)-2 primary antibody (Servicebio AF5105); immunohistochemical IL-6 primary antibody (Servicebio DF6087); RIPA lysate (Servicebio G2002-100ML); WB TNFα primary antibody (Servicebio GB12188-100); WB IL-2 primary antibody (Servicebio GB11114-100); WB IL-6 primary antibody (Servicebio GB11117-100); HRP goat anti-rabbit secondary antibody (Servicebio GB23303); HRP goat anti-mouse secondary antibody (Servicebio GB23301); prestained protein marker VII (Servicebio G2087-250UL); RNA extraction solution (Servicebio G3013); isopropanol (Sinopharm Chemical Reagent Co., Ltd., 80109218); absolute ethanol (Sinopharm Chemical Reagent Co., Ltd., 10009218); RNA lysis solution (Servicebio G3029); SweScript All-in-One RT SuperMix for qPCR (Servicebio G3337); 2 × universal blue SYBR Green qPCR Master Mix (Servicebio G3326).

Techniques: Control, Expressing

Journal: Molecular and Cellular Biology

Article Title: Muscle-Specific Deletion of the Glut4 Glucose Transporter Alters Multiple Regulatory Steps in Glycogen Metabolism

doi: 10.1128/mcb.25.21.9713-9723.2005

Figure Lengend Snippet: FIG. 4. Basal IRS-1-associated PI3K (A), Akt (B), and GSK3 (C) activity levels in muscle from WT and muscle-G4KO mice. Mice were fasted overnight. (A) PI3K activities were measured in IRS-1 immunoprecipitates and were quantitated using a PhosphorImager. (B) Muscle lysates (500 g) were subjected to immunoprecipitation with an Akt antibody that recognizes both Akt1 and Akt2. The immune pellets were assayed for kinase activity using Crosstide as the substrate. (C) Muscle lysates (500 g) were subjected to immunoprecipitation with a GSK3-specific antibody. The immune pellets were assayed for kinase activity using phospho-glycogen synthase 1 as the substrate. Results are means SEM for four to six mice per group. *, P 0.05 versus WT.

Article Snippet: The membranes were blocked with 5% nonfat dry milk for 1 h at room temperature and incubated with the following antibodies in 1% nonfat dry milk overnight at 4°C: a polyclonal Glut4 antibody (gift from H. Haspel, Henry Ford Hospital, Detroit, MI), a polyclonal insulin receptor antibody (Santa Cruz Biotechnology, Santa Cruz, CA), a polyclonal IRS-1 antibody (gift from M. White, Joslin Diabetes Center), a polyclonal antibody against the p85 subunit or p110 subunit of PI3K (Upstate Biotechnology), a polyclonal antibody for Akt that recognizes both Akt1 and Akt2 (Upstate Biotechnology), a monoclonal antibody for GSK3 that recognizes both GSK3 and GSK3 (Upstate Biotechnology), a polyclonal glycogen synthase antibody (gift from J. Lawerence, University of Virginia), a polyclonal mitogen-activated protein kinase (MAPK) antibody (gift from J. Blenis, Harvard Medical School), a polyclonal PP1 antibody (Upstate Biotechnology), a polyclonal RGL/GM antibody (gift from A. DePaoli-Roach, Indiana University) a polyclonal PTG antibody (gift from M. Brady, University of Chicago), a polyclonal phosphoglycogen synthase (Ser641) antibody (Cell Signaling, Beverly, MA), or a polyclonal hexokinase II (Chemicon, Temecula, CA).

Techniques: Activity Assay, Immunoprecipitation

Journal: Molecular and Cellular Biology

Article Title: Muscle-Specific Deletion of the Glut4 Glucose Transporter Alters Multiple Regulatory Steps in Glycogen Metabolism

doi: 10.1128/mcb.25.21.9713-9723.2005

Figure Lengend Snippet: FIG. 5. Insulin-stimulated IRS-1-associated PI3K (A), Akt (B), GSK3 (C), and glycogen synthase (D) activity levels in muscle from WT and muscle-G4KO mice. After an overnight fast, mice were injected i.v. with saline (, white bars) or 10 U/kg insulin (, black bars). Three min later, muscle was removed. (A) PI3K activities were measured in IRS-1 immunoprecipitates and were quantitated using a PhosphorImager. (B) Muscle lysates (500 g) were subjected to immunoprecipitation with an Akt antibody that recognizes both Akt1 and Akt2. The immune pellets were assayed for kinase activity using Crosstide as the substrate. (C) Muscle lysates (500 g) were subjected to immunoprecipitation with a GSK3- specific antibody. The immune pellets were assayed for kinase activity using phospho-glycan synthase 1 as the substrate. (D) The ratio of glycogen synthase (GS) activity represents the activity measured in the absence divided by that in the presence of glucose-6-phosphate. Results are means SEM for four to six mice per group. *, P 0.05 versus insulin-stimulated WT.

Article Snippet: The membranes were blocked with 5% nonfat dry milk for 1 h at room temperature and incubated with the following antibodies in 1% nonfat dry milk overnight at 4°C: a polyclonal Glut4 antibody (gift from H. Haspel, Henry Ford Hospital, Detroit, MI), a polyclonal insulin receptor antibody (Santa Cruz Biotechnology, Santa Cruz, CA), a polyclonal IRS-1 antibody (gift from M. White, Joslin Diabetes Center), a polyclonal antibody against the p85 subunit or p110 subunit of PI3K (Upstate Biotechnology), a polyclonal antibody for Akt that recognizes both Akt1 and Akt2 (Upstate Biotechnology), a monoclonal antibody for GSK3 that recognizes both GSK3 and GSK3 (Upstate Biotechnology), a polyclonal glycogen synthase antibody (gift from J. Lawerence, University of Virginia), a polyclonal mitogen-activated protein kinase (MAPK) antibody (gift from J. Blenis, Harvard Medical School), a polyclonal PP1 antibody (Upstate Biotechnology), a polyclonal RGL/GM antibody (gift from A. DePaoli-Roach, Indiana University) a polyclonal PTG antibody (gift from M. Brady, University of Chicago), a polyclonal phosphoglycogen synthase (Ser641) antibody (Cell Signaling, Beverly, MA), or a polyclonal hexokinase II (Chemicon, Temecula, CA).

Techniques: Activity Assay, Injection, Saline, Immunoprecipitation, Glycoproteomics

Journal: Molecular and Cellular Biology

Article Title: Muscle-Specific Deletion of the Glut4 Glucose Transporter Alters Multiple Regulatory Steps in Glycogen Metabolism

doi: 10.1128/mcb.25.21.9713-9723.2005

Figure Lengend Snippet: FIG. 9. Signaling protein levels in muscle from WT and muscle- G4KO mice. Mice were fasted overnight. Proteins in muscle lysates (25 to 50 g) were separated by SDS-PAGE on 6, 8, or 10% gels and transferred to nitrocellulose membranes. Insulin receptor (IR), IRS-1, p110, p85, Akt, GSK3/, and MAPK were visualized by immuno- blotting with specific antibodies. This blot is representative of three blots on six to nine mice per group.

Article Snippet: The membranes were blocked with 5% nonfat dry milk for 1 h at room temperature and incubated with the following antibodies in 1% nonfat dry milk overnight at 4°C: a polyclonal Glut4 antibody (gift from H. Haspel, Henry Ford Hospital, Detroit, MI), a polyclonal insulin receptor antibody (Santa Cruz Biotechnology, Santa Cruz, CA), a polyclonal IRS-1 antibody (gift from M. White, Joslin Diabetes Center), a polyclonal antibody against the p85 subunit or p110 subunit of PI3K (Upstate Biotechnology), a polyclonal antibody for Akt that recognizes both Akt1 and Akt2 (Upstate Biotechnology), a monoclonal antibody for GSK3 that recognizes both GSK3 and GSK3 (Upstate Biotechnology), a polyclonal glycogen synthase antibody (gift from J. Lawerence, University of Virginia), a polyclonal mitogen-activated protein kinase (MAPK) antibody (gift from J. Blenis, Harvard Medical School), a polyclonal PP1 antibody (Upstate Biotechnology), a polyclonal RGL/GM antibody (gift from A. DePaoli-Roach, Indiana University) a polyclonal PTG antibody (gift from M. Brady, University of Chicago), a polyclonal phosphoglycogen synthase (Ser641) antibody (Cell Signaling, Beverly, MA), or a polyclonal hexokinase II (Chemicon, Temecula, CA).

Techniques: SDS Page

Journal: Nature communications

Article Title: Primate-Specific miR-576-3p Sets Host Defense Signaling Threshold

doi: 10.1038/ncomms5963

Figure Lengend Snippet: a , Schematic showing the region of the 3’ UTR of STING that is recognized by the miR-576-3p seed sequence. b , HBEC were transfected with control or miR-576-3p (M-576-3p) mimic for 72 h. Cells were then mock infected or infected with VSV-GFP at an MOI 3 for 3 h. Total RNA was harvested from cells and STING mRNA levels were determined by qPCR and normalized to levels of β-actin. c , HBEC were transfected with miRNA mimic (M-576-3p) or inhibitor (I-576-3p) as in b and mock-infected or infected with VSV-GFP at MOI 0.1 for 18 h. Cell lysates were harvested and subjected to western blot analysis with anti-STING antibodies. β-actin serves as loading control. d,e , HBEC were transfected with 1 µg/ml of poly (I:C) for 6 h ( d ) or treated with 100 U/ml of IFNβ for 18 h ( e ) and levels of pri-mir-576 and SEC24B mRNA were analyzed by qPCR. f,g , HBEC were transfected with poly (I:C) as in d after siRNA knockdown of NFκB (P65) or IRF3. Relative pri-mir-576 ( f ) or SEC24B mRNA ( g ) levels were measured by qPCR. h , HBEC expressing control or miR-576-3p mimic (M-576-3p) or an siRNA targeting STING were transfected with poly (I:C) and levels of IFNβ mRNA were measured by qPCR. i , HBEC were transfected with control or miR-576-3p mimic (M-576-3p) and pre-treated with 100 U/ml of IFNβ prior to VSV infection. Cell viability was determined by measuring ATP levels of mock or infected cells. j , HBEC expressing control or miR-576-3p inhibitor (I-576-3p) were transfected with poly (I:C) and levels of IFNβ were measured by qPCR. Data are representative of three independent experiments. Unpaired two tailed t-test was used and error bars represent SD. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: Recombinant human IFNβ was obtained from R&D systems.

Techniques: Sequencing, Transfection, Control, Infection, Western Blot, Knockdown, Expressing, Two Tailed Test

Journal: Nature communications

Article Title: Primate-Specific miR-576-3p Sets Host Defense Signaling Threshold

doi: 10.1038/ncomms5963

Figure Lengend Snippet: a , HBEC were transfected with 1 µg/ml of poly (I:C) for 6 h or treated with 100 U/ml of IFNβ for 18 h and levels of mature miR-576-3p were determined by qPCR. b , HBEC were mock infected or infected with HSV-1 at MOI 10 for 3 h and levels of miR-576-3p were measured by qPCR. Unpaired two tailed t-test was used and error bars represent SD. *p<0.05, **p<0.01. ns, non significant with respect to control.

Article Snippet: Recombinant human IFNβ was obtained from R&D systems.

Techniques: Transfection, Infection, Two Tailed Test, Control

Journal: Nature communications

Article Title: Primate-Specific miR-576-3p Sets Host Defense Signaling Threshold

doi: 10.1038/ncomms5963

Figure Lengend Snippet: a , Model illustrates regulation of IFN expression by miR-576-3p as a feedback mechanism. Targets of miR-576-3p are indicated by inhibitory red lines. b , Public available microRNA array datasets (GSE37425 and GSE37426) of synovial or renal tissues from RA or SLE patients, respectively, along with controls were obtained from the GEO database . Normalized expression values of miR-576-3p from each sample were used to calculate the relative levels and p-values of miR-576-3p for SLE or RA patients as compared to controls. Unpaired two tailed t-test was used and error bars represent SD. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: Recombinant human IFNβ was obtained from R&D systems.

Techniques: Expressing, Two Tailed Test