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: Inflammation and Regeneration

Article Title: Novel artificial nerve transplantation of human iPSC-derived neurite bundles enhanced nerve regeneration after peripheral nerve injury

doi: 10.1186/s41232-024-00319-4

Figure Lengend Snippet: Early-stage tissue evaluation after neurite bundle-derived artificial nerve transplantation. a , b Immunohistochemistry images with anti-NFH and anti-CD31 antibodies. Dotted arrows indicate the proximal side of the intact nerve areas, and solid arrows indicate the NFH- and CD31-positive regeneration tip in the reconstructed tissue. Scale bars = 1 mm. c , d Quantitative comparison of axonal extension distance was indicated by NFH-positive areas and that of vascular elongation distance was demonstrated by CD31-positive areas at 1 and 2 weeks after transplantation. Significant differences between groups elongation distances were observed in 2-week samples only. e , f Iba1 immunohistochemistry image of the proximal part of the regenerated tissue at 2 weeks after transplantation (e). Scale bars = 1 mm. Quantitative analysis of Iba1-positive areas demonstrated that a larger number of macrophages aggregated in the Auto, motor TP and sensory TP (f). ( n = 3). * p < 0.05, ** p < 0.01, N.S. = not significant. Data are represented as the mean ± SEM

Article Snippet: The primary antibodies used in this study were human polyclonal anti-pan-ELAVL (ELAV-like protein 2/3/4) antibody (1:2000, kindly provided from Prof. Robert Darnell, Rockefeller University), rabbit polyclonal anti-TrkA antibody (1:500, kindly provided from Prof. Louis F. Reichardt, UCSF), chicken polyclonal anti-TrkB antibody (1:500, kindly provided from Prof. Louis F. Reichardt, UCSF), goat polyclonal anti-TrkC antibody (AF373, 1:200, R&D systems), rabbit polyclonal anti-parvalbumin antibody (PV-28, 1:1000, Swant), rabbit polyclonal anti-CGRP antibody (BML-CA1134, 1:500, Enzo Life Sciences), Goat polyclonal anti-Choline acetyltransferase antibody (AB144P, 1:200, Chemicon), Mouse monoclonal anti-Islet1 and Islet2 antibody (39.4D5, 1:200, DSHB), rabbit polyclonal anti-Neurofilament heavy polypeptide antibody (ab8135, 1:500, Abcam), goat polyclonal anti-mouse and anti-rat CD31 (AF3628, 1:100, R&D), rabbit polyclonal anti-Iba1 antibody (GtX100042, 1:500, GeneTex), goat anti-GFP antibody (600–101-215, 1:1000, Rockland).

Techniques: Derivative Assay, Transplantation Assay, Immunohistochemistry, Comparison

Journal: medRxiv

Article Title: In severe COVID-19, SARS-CoV-2 induces a chronic, TGF-β-dominated adaptive immune response

doi: 10.1101/2020.09.04.20188169

Figure Lengend Snippet: Peripheral blood activated B cells (CD27 high CD38 high ) were isolated and sorted by FACS for single cell sequencing. a) Percentage of CD27 high CD38 high cells among live B cells. Each sample is represented by a circle. The line indicates the median. Unpaired two sided Mann-Whitney U test. b) UMAP representation of 72277 CD27 high CD38 high sorted B cells from 9 COVID-19 ICU patients and 3 healthy controls. Between 2416 and 11229 cells were recovered per sample. Transcriptionally similar clusters were identified using shared nearest neighbor (SNN) modularity optimization (left). The combination of CD19, MS4A1, CD27, CD38, IFIT1, MIKI67, IRF4 and PRDM1 expression were used for annotation of the different activation/differentiation stages (right). c) Heatmap of genes that resemble markers for the six different clusters. Depicted are genes with a p-value < 0.01 (Wilcoxon rank sum test) after bonferroni correction, and an average absolute fold-change > log2(1.3). Shown are z-scores of the average expression. d) UMAP representation of the expression levels of selected signature genes for activated/differentiated B cells. e) UMAP representation of analyzed cells from one healthy control and two ICU patients, representing early (first week, patient #1) and late (>7days, patient #5) phase after ICU admission. f) Percentage of cells belonging to a defined cluster among all sequenced B cells per donor. Each dot represents one time point from one single donor. Donors were grouped as “HC” for healthy controls (n = 3), “1st week” for patients within 7 days after ICU admission (n = 6) and “Late” for patients who have been admitted to the ICU for more than a week at the time of analysis (n = 8). The line indicates the median. Significance was determined by using a two-sided analysis of varience (ANOVA) folowed by Tukey’s multiple comparison test with corrected p values as indicated in the figure.

Article Snippet: Fluorescence-coupled antibodies used: IgA2-PE (Clone REA995, Miltenyi, Cat. No.130-117-763, dilution 1:50); CD27-PE (Clone REA499, Miltenyi, Cat. No. 130-114-166, dilution 1:50); CD38-PE (Clone IB6, Miltenyi, Cat. No. 130-113-427, dilution 1:50) and IgA-PE (Clone IS11–8E10, Miltenyi, Cat. No. 130-114-002, dilution 1:50).

Techniques: Isolation, Sequencing, MANN-WHITNEY, Expressing, Activation Assay, Control, Comparison

Journal: medRxiv

Article Title: In severe COVID-19, SARS-CoV-2 induces a chronic, TGF-β-dominated adaptive immune response

doi: 10.1101/2020.09.04.20188169

Figure Lengend Snippet: a) 5-marker MELC panel of SARS-CoV2-positive and control lungs (SARS-CoV2-negative). Respective patient characteristics are given in supplementary Fig. 4a. Each image of each patient depicts the same field of view of the same section, sequentially stained with the fluorescence-labelled antibodies indicated and the nuclear stain DAPI. Magenta arrows indicate IgA2+IgA+CD27 + CD38 + cells (containing a nucleus). Images contain 2048 × 2048 pixels and are generated using an inverted wide-field fluorescence microscope with a 20x objective, a lateral resolution of 325 nm and an axial resolution above 5 µm. Scale bar: 100 µm. b) Absolute numbers of IgA2+IgA+CD27 + CD38 + cells per field of view in all MELC runs acquired (two runs per patient except for COVID-19_A with four runs and Control_B with one single run; see and supplementary Fig. 4b). Each field of view is represented by a circle. The line indicates the median. Unpaired two sided Mann-Whitney U test. c) Region of interest of one exemplary control and COVID-19 lung (as in a)), showing an overlay of the indicated markers. White arrows point out IgA2+CD27 + CD38 + cells. Scale bar: 20 µm. (d-e) Bronchoalveolar lavage (BAL) cells for single cell sequencing were enriched for CD45 + cells via MACS and live cells were further sorted using FACS (see supplementary Fig. 4c). d) UMAP of 5459 cells representing clusters containing T cells, B cells and CD14 -expressing cells from patient #1 on day 59 following ICU admission. B and T cell cluster identification based on BCR/TCR, CD19, CD3E, CD4 and CD8A expression (see supplementary Fig. 4d). UMAP representation of expression of TGFB1, IL21, CD40LG and IFNG . e) UMAP coordinates and clustering was computed for 433 and 2723 cells from patient #9 on days 31 and 46 following ICU admission, respectively. UMAP representation of expression of TGFB1, IL21, CD40LG and IFNG at the two different time points is shown side by side.

Article Snippet: Fluorescence-coupled antibodies used: IgA2-PE (Clone REA995, Miltenyi, Cat. No.130-117-763, dilution 1:50); CD27-PE (Clone REA499, Miltenyi, Cat. No. 130-114-166, dilution 1:50); CD38-PE (Clone IB6, Miltenyi, Cat. No. 130-113-427, dilution 1:50) and IgA-PE (Clone IS11–8E10, Miltenyi, Cat. No. 130-114-002, dilution 1:50).

Techniques: Marker, Control, Staining, Fluorescence, Generated, Microscopy, MANN-WHITNEY, Sequencing, Expressing