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: Human reproduction (Oxford, England)

Article Title: Interleukin 1 regulates its own receptors in human endometrial cells via distinct mechanisms.

doi: 10.1093/humrep/dep192

Figure Lengend Snippet: Figure 1 Effect of IL1B on mbIL1R, mbIL1R2 and IL1RA expression in KLE cells. Confluent KLE cell cultures were treated with 0.1 ng/ml IL1B for varying periods of time (0–24 h) (A) or with different concentrations of IL1B (0, 0.1, 1 and 10 ng/ml) for 12 h (B). Cells were recovered to evaluate IL1R1, IL1R2 and IL1RA protein expression in total cell protein extracts by western blot. a-Tubulin was also probed on the same membranes to ensure equal protein loading. The intensity of mbIL1Rs, IL1RA and corresponding a-tubulin bands was evaluated by densitometric analysis (C and D). Values were normalized to a-tubulin band intensity and expressed as % of control (ratio of normalized mbIL1R1, mbIL1R2 or IL1RA band intensity detected following treatment with IL1B to that detected following incubation with the control culture medium alone for an equivalent period of time). *P , 0.05 and **P , 0.01 as compared with control. Data are from three different experiments. MM, minimal medium; mb, membrane-bound; s, soluble.

Article Snippet: IL1R2 ELISA is based on the use of a mouse monoclonal anti-human IL1R2 antibody for capture and a goat polyclonal anti-human IL1R2 antibody for detection (R&D systems).

Techniques: Expressing, Western Blot, Control, Incubation, Membrane

Journal: Human reproduction (Oxford, England)

Article Title: Interleukin 1 regulates its own receptors in human endometrial cells via distinct mechanisms.

doi: 10.1093/humrep/dep192

Figure Lengend Snippet: Figure 3 Immunocytofluorescence of IL1R1, IL1R2 and IL1RA in KLE cells. Cells cultured in chamber slides were incubated for 24 h with the culture medium alone or containing 0.1 ng/ml IL1B. Detection of IL1R1 (A and C), IL1R2 (E and G) and IL1RA (I and K) was performed by immunocytochemical staining using specific monoclonal mouse antibodies. Note the increase in IL1R1 (C), IL1R2 (G) and IL1RA (K) immunofluorescent signal in KLE cells exposed to IL1B by comparison with non-stimulated cells (A, E and I, respectively). No immunofluorescence was observed in the absence of primary antibodies (B, F and J) or the presence of mouse IgGs (D, H and L) (controls). Data are representative of four different experiments.

Article Snippet: IL1R2 ELISA is based on the use of a mouse monoclonal anti-human IL1R2 antibody for capture and a goat polyclonal anti-human IL1R2 antibody for detection (R&D systems).

Techniques: Cell Culture, Incubation, Staining, Comparison

Journal: Human reproduction (Oxford, England)

Article Title: Interleukin 1 regulates its own receptors in human endometrial cells via distinct mechanisms.

doi: 10.1093/humrep/dep192

Figure Lengend Snippet: Figure 4 Effect of IL1B on IL1R1, IL1R2 and IL1RA mRNA expression in KLE cells. Confluent KLE cell cultures were treated with 0.1 ng/ml IL1B for varying periods of time (0–24 h) (A, B and C, respectively) or with different con- centrations of IL1B (0, 0.1, 1 and 10 ng/ml) for 12 h (D, E and F, respectively). Total RNA was extracted and reverse transcribed, IL1R1, IL1R2, IL1RA and GAPDH cDNAs were amplified by Real-Time PCR as described in Materials and Methods and IL1R1, IL1R2 and IL1RA mRNA levels were normalized to GAPDH mRNA levels. Data were expressed as % of control (ratio of IL1R1, IL1R2 or IL1RA mRNA levels found in cells incubated with IL1 to those found in cells incubated with the control basal culture medium for an equivalent period of time). *P , 0.05, **P , 0.01 and ***P , 0.001 as compared with control. Data are from three different experiments.

Article Snippet: IL1R2 ELISA is based on the use of a mouse monoclonal anti-human IL1R2 antibody for capture and a goat polyclonal anti-human IL1R2 antibody for detection (R&D systems).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Incubation

Journal: Human reproduction (Oxford, England)

Article Title: Interleukin 1 regulates its own receptors in human endometrial cells via distinct mechanisms.

doi: 10.1093/humrep/dep192

Figure Lengend Snippet: Figure 5 Effect of IL1B on IL1R1, IL1R2 and IL1RA mRNA stability and gene expression in KLE cells. Confluent KLE cell cultures were stimulated with IL1B (1 ng/mll) for 12 h. Actinomycin D (10 mg/ml) was added to stop the de novo RNA synthesis, and cells were harvested after 0, 2, 4 and 8 h of incubation with actinomycin D. Total RNA was extracted and reverse transcribed. cDNA was analyzed by real-time PCR with specific primers for IL1R1, IL1R2, IL1RA and GAPDH. Levels of IL1R1, IL1R2 and IL1RA mRNA were normalized to those of GAPDH to assess the kinetics of IL1R1 (A), IL1R2 (B) and IL1RA (C) mRNA degradation. Data were expressed as % of control (ratio of IL1R1, IL1R2 or IL1RA mRNA levels found at different periods of time following the arrest of de novo mRNA transcription to that found at the time of arrest); data are from three different experiments. To evaluate IL1R1, IL1R2 and IL1RA transcriptional activation (D), confluent KLE cell cultures were stimulated with IL1B (1 ng/ml) for 6 h, cell nuclei were isolated and nuclear mRNA transcription was analyzed by nuclear run-on as described in Materials and Methods. DNA samples immobilized onto nylon membranes were as follows: lane 1, IL1R1 cDNA; lane 2, IL1R2 cDNA; lane 3, IL1RA; lane 4, 28S cDNA; and lane 5, pBluescript plasmid DNA. Radioactive transcripts were from KLE cells stimulated in MM or with 1 ng/ml IL1B. Data are repre- sentative of four different experiments.

Article Snippet: IL1R2 ELISA is based on the use of a mouse monoclonal anti-human IL1R2 antibody for capture and a goat polyclonal anti-human IL1R2 antibody for detection (R&D systems).

Techniques: Gene Expression, Incubation, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Activation Assay, Isolation, Plasmid Preparation

Journal: Human reproduction (Oxford, England)

Article Title: Interleukin 1 regulates its own receptors in human endometrial cells via distinct mechanisms.

doi: 10.1093/humrep/dep192

Figure Lengend Snippet: Figure 6 Effect of IL1B on mbIL1R1, mbIL1R2 and IL1RA expression in endometrial epithelial cells. Confluent endometrial epithelial cell cultures were treated with different concentrations of IL1B (0, 0.1, 1 and 10 ng/ml) for 12 h. Cells were recov- ered to evaluate IL1R1, IL1R2 and IL1RA protein expression in total cell protein extracts by Western blot (A). a-Tubulin was probed on the same membranes to ensure equal protein loading. Culture supernatants were recovered to evaluate sIL1R2 release (C) and IL1RA secretion (D) by ELISA; data were from four different endometrial samples and expressed as % of control (ratio of sIL1R2 or IL1RA concentrations detected in the presence of IL1 to those detected in the control basal culture medium for an equivalent period of 12 h). *P , 0.05, as compared with control. The intensity of mbIL1Rs, IL1RA and corresponding a-tubulin bands was evaluated by densitometric analysis (B). Values were normalized to a-tubulin band intensity and expressed as % of control (ratio of normalized mbIL1R1, mbIL1R2 or IL1RA band intensity detected following treatment with IL1B to that detected following incubation with the control culture medium alone for an equivalent period of time). *P , 0.05 and **P , 0.01 as compared with control. Data were from three different endometrial samples. sIL1R1 and sIL1R2 release and IL1RA secretion from endometrial epithelial cell cultures was also analyzed by metabolic labeling with 35S-cysteine and immunoprecipitation using a goat anti-IL1R1, goat anti-IL1R2 or rabbit anti- IL1RA antibody (E). Immunoprecipitated proteins were then analyzed by SDS-PAGE as described in Materials and Methods (representative data from three different endometrial samples); mb, membrane-bound; s, soluble.

Article Snippet: IL1R2 ELISA is based on the use of a mouse monoclonal anti-human IL1R2 antibody for capture and a goat polyclonal anti-human IL1R2 antibody for detection (R&D systems).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Control, Incubation, Labeling, Immunoprecipitation, SDS Page, Membrane

Journal: Human reproduction (Oxford, England)

Article Title: Interleukin 1 regulates its own receptors in human endometrial cells via distinct mechanisms.

doi: 10.1093/humrep/dep192

Figure Lengend Snippet: Figure 7 Effect of IL1B on IL1R1, IL1R2 and IL1RA mRNA expression in endometrial epithelial cells. Confluent cultures were treated with 0.1 ng/ml IL1B with different concentrations of IL1B (0, 0.1, 1 and 10 ng/ml) for 12 h. Total RNA was extracted and reverse transcribed, IL1R1, IL1R2, IL1RA and GAPDH cDNAs were amplified by Real-Time PCR as described in Materials and Methods and IL1R1 (A), IL1R2 (B) and IL1RA (C) mRNA levels were normalized to GAPDH mRNA levels. Data were from three different endometrial samples and expressed as % of control (ratio of IL1R1, IL1R2 or IL1RA mRNA levels found in cells incubated with IL1 to those found in cells incubated with the control basal culture medium for an equivalent period of 12 h). *P , 0.05 and **P , 0.01 as compared with control.

Article Snippet: IL1R2 ELISA is based on the use of a mouse monoclonal anti-human IL1R2 antibody for capture and a goat polyclonal anti-human IL1R2 antibody for detection (R&D systems).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Incubation

Journal: Genes and immunity

Article Title: Despite IFN-lambda receptor expression, blood immune cells, but not keratinocytes or melanocytes, have an impaired response to type III interferons: implications for therapeutic applications of these cytokines.

doi: 10.1038/gene.2009.72

Figure Lengend Snippet: Figure 4 Peripheral blood mononuclear cells (PBMCs), keratino- cytes and HepG2 cells express interferon (IFN)-lR1 and IL-10R2 protein. Protein expressions of IFN-l receptor components and of the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were analyzed in the cell lysates of PBMCs from three donors as well as of keratinocytes and HepG2 cells using the western blot technique.

Article Snippet: For the detection of IFN-l receptor components, we incubated blots with anti-IFNlR1 pAbs (50 ng ml 1; Sigma-Aldrich), anti-IL-10R2 pAbs (600 ng ml 1; R&D Systems) and anti-GAPDH mAb (200 ng ml 1; clone 6C5; Millipore), followed by peroxidase-conjugated AffiniPure goat anti-rabbit IgG (H and L), rabbit anti-goat IgG (H and L) and goat antimouse IgG (H and L) (all from Dianova) incubation, respectively, and final ECL detection.

Techniques: Western Blot