Journal: Cell

Article Title: Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches

doi: 10.1016/j.cell.2021.12.018

Figure Lengend Snippet:

Article Snippet: Anti-Human S100A8 (REA917) PE , Miltenyi Biotec , 130-115-253; RRID: AB_2726963.

Techniques: Purification, Recombinant, Staining, cDNA Synthesis, Gene Expression, Software, Microscopy

Journal: Respiratory research

Article Title: Deficiency of S100A8/A9 attenuates pulmonary microvascular leakage in septic mice.

doi: 10.1186/s12931-023-02594-0

Figure Lengend Snippet: Fig. 1 Elevated S100 calcium-binding protein (S100) A8/A9 levels are associated with pulmonary vascular hyperpermeability and disease severity in polymicrobial sepsis. (A) S100A8/A9, occludin, and VE-cadherin protein expression levels in the lungs of WT mice were assessed using Western blotting 12 h after CLP surgery. The statistical results of S100A8/A9, occludin, and VE-cadherin are illustrated in (B), (C), and (D) respectively (n = 5). (E) The body temperatures were assessed using a rectal thermometer 12 h after CLP/sham surgery (n = 6). (F) Survival rates among the WT and S100A9 KO mice after CLP surgery were compared using the Kaplan–Meier test (n = 15, three independent experiments). (G) Carotid pressures were measured 12 h after surgery and the related statistical results are presented in (H-I) (n = 5). S100, S100 calcium-binding protein; VE, vascular endothelial; WT, wide-type. KO, knockout. CLP, cecal ligation and puncture. VE-cad, VE-cadherin. The data are presented as the mean ± standard error of the mean. ns, no significant dif ference. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001

Article Snippet: Subsequently, the cells were treated with 2.0 μg/mL of human recombinant S100A8/A9 (Cat #HY-P71076, MedChemexpress, US) or PBS for 2 h, as reported previously [28].

Techniques: Binding Assay, Expressing, Western Blot, Knock-Out, Ligation

Journal: Respiratory research

Article Title: Deficiency of S100A8/A9 attenuates pulmonary microvascular leakage in septic mice.

doi: 10.1186/s12931-023-02594-0

Figure Lengend Snippet: Fig. 2 S100A9 knockout attenuates CLP-induced pulmonary inflammation, vascular leakage, and acute lung injury. (A) The S100A8/A9 protein expres sion in the lungs of mice was determined using Western blotting 12 h after CLP surgery (n = 6). The statistical results are presented in (B). (C-E) The TNF-α (C), IL-1β (D), and MCP-1 (E) mRNA levels in the lungs of mice were analysed using RT-PCR 12 h after CLP surgery (n = 6). (F) The levels of EB inundated the lung tissues in mice after CLP surgery (n = 6). The statistical results of lung EB absorbance at 610 nm are presented in (G). (H) The lung injury score was evaluated using a scoring system, as described in the Methods (n = 6). The statistical results of the histological score were exposed in (I). S100, S100 calcium-binding protein; CLP, cecal ligation and puncture; TNF, tumour necrosis factor; IL, interleukin; MCP-1, monocyte chemoattractant protein; mRNA, messenger ribonucleic acid; RT-PCR, reverse transcription polymerase chain reaction; EB, Evans blue. The data are presented as the mean ± standard error of the mean. ns, no significant difference. *P < 0.05; **P < 0.01; ****P < 0.0001

Article Snippet: Subsequently, the cells were treated with 2.0 μg/mL of human recombinant S100A8/A9 (Cat #HY-P71076, MedChemexpress, US) or PBS for 2 h, as reported previously [28].

Techniques: Knock-Out, Western Blot, Reverse Transcription Polymerase Chain Reaction, Binding Assay, Ligation, Reverse Transcription, Polymerase Chain Reaction

Journal: Respiratory research

Article Title: Deficiency of S100A8/A9 attenuates pulmonary microvascular leakage in septic mice.

doi: 10.1186/s12931-023-02594-0

Figure Lengend Snippet: Fig. 5 S100A8/A9 promotes endothelial barrier dysfunction in HUVECs. (A) Western blot analysis of occludin, VE-cadherin, p-ERK, p-p38, Bax, and Bcl-2 expressions in HUVECs 2 h after rhS100A8/A9 administration (n = 6). The statistical results are presented in (B-F). S100, S100 calcium-binding protein; HUVECs, human umbilical vein endothelial cells; VE, vascular endothelial; p-ERK, phosphorylated extracellular signal-regulated kinase; p-p38, phosphor ylated-p38; B-cell leukaemia/lymphoma 2 protein (Bcl-2)-associated X protein; rhS100A8/A9, recombinant S100A8/A9. The data are presented as the mean ± standard error of the mean. *P < 0.05; ***P <0.001; ****P < 0.0001

Article Snippet: Subsequently, the cells were treated with 2.0 μg/mL of human recombinant S100A8/A9 (Cat #HY-P71076, MedChemexpress, US) or PBS for 2 h, as reported previously [28].

Techniques: Western Blot, Binding Assay, Recombinant

Journal: International Journal of Molecular Sciences

Article Title: Hypoxia-Induced S100A8 Expression Activates Microglial Inflammation and Promotes Neuronal Apoptosis

doi: 10.3390/ijms22031205

Figure Lengend Snippet: Hypoxia increased the production of S100 calcium-binding protein A8 (S100A8) in neuron and microglia and induced the release of S100A8 in SH-SY5Y cells. ( A , B ) S100A8 expression (red) were detected by immunocytochemical analysis in primary cultured neurons (NeuN, neuron marker) and cultured mixed glia (Iba1, microglial marker and GFAP, astrocyte marker) exposed to hypoxic conditions for 48 h. Scheme 25 μm. S100A8 expression was detected by western blot analysis in ( C , D ) SH-SY5Y cells and ( E , F ) BV-2 cells exposed to hypoxic conditions for 48 h. ( G , H ) S100A8 protein expression in BV-2 cells were confirmed by immunocytochemistry and ( I ) S100A8 release in SH-SY5Y was measured by enzyme-linked immunosorbent assay (ELISA) at 48 h after hypoxia. Values of * p < 0.05, ** p < 0.01, *** p < 0.001 versus control were considered as statistically significant.

Article Snippet: S100A8, TNF-α, IL-6, IL-1β and PGE2 were quantitatively measured by an enzyme-linked immunosorbent assay (ELISA) using the human S100A8 Duoset ELISA kits, the mouse TNF-α, IL-6 and IL-1β DuoSet ELISA kits, and the PGE2 parameter assay kit (R&D systems, Minneapolis, MN, USA), according to the manufacturer’s instructions.

Techniques: Binding Assay, Expressing, Cell Culture, Marker, Western Blot, Immunocytochemistry, Enzyme-linked Immunosorbent Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Hypoxia-Induced S100A8 Expression Activates Microglial Inflammation and Promotes Neuronal Apoptosis

doi: 10.3390/ijms22031205

Figure Lengend Snippet: S100A8 induces pro-inflammatory cytokines and inflammation in BV-2 cells. BV-2 cells were stimulated with S100A8 (10 μg/mL) for 24 h. ( A ) The supernatant was collected and TNF-α and interleukin-6 (IL-6) analyzed by ELISA. ( B ) The protein and mRNA were extracted, and the expression levels of IL-1β were assessed by ELISA and RT-qPCR. ( C – E ) The protein was extracted, separated on 10% SDS-acrylamide gels (15 μg/lane) and transferred to nitrocellulose membrane. The protein expression level was detected by western blotting with anti-ERK1/2, anti-phospho-ERK1/2 (p-ERK1/2), anti-JNK and anti-p-JNK. ( F ) Cells were pre-treated with ERK inhibitor (PD98059, 20 μM), JNK inhibitor (SP600125, 10 μM) or the equivalent volume of DMSO for 1 h, then stimulated for 24 h with LPS or S100A8 for ELISA of TNF-α, IL-6. Data from three independent experiments are presented as the means ± S.D. Values of * p < 0.05, *** p < 0.001 versus control; ### p < 0.001 versus S100A8-treated sample were considered as statistically significant.

Article Snippet: S100A8, TNF-α, IL-6, IL-1β and PGE2 were quantitatively measured by an enzyme-linked immunosorbent assay (ELISA) using the human S100A8 Duoset ELISA kits, the mouse TNF-α, IL-6 and IL-1β DuoSet ELISA kits, and the PGE2 parameter assay kit (R&D systems, Minneapolis, MN, USA), according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Membrane, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Hypoxia-Induced S100A8 Expression Activates Microglial Inflammation and Promotes Neuronal Apoptosis

doi: 10.3390/ijms22031205

Figure Lengend Snippet: S100A8 induces inflammasome priming by toll-like receptor (TLR)-4 receptors associated with ERK and JNK pathway in BV-2 cells. BV-2 cells were incubated for 24 h with LPS (1 μg/mL) or S100A8 (10 μg/mL) followed by Adenosine 5′-triphosphate disodium salt hydrate (ATP) (1 mM) for 1 h. ( A , B ) The NLRP3, ASC, and ( C , D ) cleaved caspase-1 were detected by western blotting. β-actin was used as an internal control. ( E , F ) BV-2 cells were lysed to whole lysates and IκB-α phosphorylation was analyzed by western blotting. ( G , H ) The translocation of nuclear factor- κB (NF-κB) was also detected by western blotting. BV-2 cells were lysed to cytosolic extracts and nucleic extracts. Lamin-B1 was used as internal controls. ( I , J ) BV-2 microglial cells were pre-treated with PD98059 (ERK inhibitor, 20 μM), SP600125 (JNK inhibitor, 10 μM), TAK-202 (TLR4 inhibitor, 10 μg/mL) or an equivalent volume of DMSO and stimulated for 24 h with LPS or S100A8. Cells harvested and lysed in RIPA buffer for western blotting of NLRP3. Results are from one experiment that is representative of at least three others. Data from three independent experiments are presented as the means ± S.D. Values of * p < 0.05, ** p < 0.01 versus control; # p < 0.05, ## p < 0.01 versus S100A8-treated sample were considered as statistically significant.

Article Snippet: S100A8, TNF-α, IL-6, IL-1β and PGE2 were quantitatively measured by an enzyme-linked immunosorbent assay (ELISA) using the human S100A8 Duoset ELISA kits, the mouse TNF-α, IL-6 and IL-1β DuoSet ELISA kits, and the PGE2 parameter assay kit (R&D systems, Minneapolis, MN, USA), according to the manufacturer’s instructions.

Techniques: Incubation, Western Blot, Control, Phospho-proteomics, Translocation Assay

Journal: International Journal of Molecular Sciences

Article Title: Hypoxia-Induced S100A8 Expression Activates Microglial Inflammation and Promotes Neuronal Apoptosis

doi: 10.3390/ijms22031205

Figure Lengend Snippet: S100A8 derived from neuronal cells induces NLRP3 inflammasome priming in microglia under hypoxic conditions. BV-2 cells were pre-treated with TAK-202 (TLR4 inhibitor, 10 μg/mL) for 1 h, then stimulated for 48 h in hypoxic condition with SH-SY5Y cells indirectly co-cultured in 0.4 μm pore transwell. ( A ) The protein expression level was detected by western blotting with NLRP3. β-actin was used as an internal control. ( B ) Quantitative analysis of NLRP3 levels. Data from three independent experiments are presented as the means ± S.D. Values of ** p < 0.01 versus control; # p < 0.05 versus co-cultured sample were considered as statistically significant.

Article Snippet: S100A8, TNF-α, IL-6, IL-1β and PGE2 were quantitatively measured by an enzyme-linked immunosorbent assay (ELISA) using the human S100A8 Duoset ELISA kits, the mouse TNF-α, IL-6 and IL-1β DuoSet ELISA kits, and the PGE2 parameter assay kit (R&D systems, Minneapolis, MN, USA), according to the manufacturer’s instructions.

Techniques: Derivative Assay, Cell Culture, Expressing, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Hypoxia-Induced S100A8 Expression Activates Microglial Inflammation and Promotes Neuronal Apoptosis

doi: 10.3390/ijms22031205

Figure Lengend Snippet: The expression of S100A8 in microglial cell induces apoptosis of neuronal cells in hypoxic condition. ( A , B ) SH-SY5Y cells incubated without or with S100A8 KD BV-2 cells for 48 h in hypoxic condition. Cleaved caspase-3 immunofluorescence images and were detected and quantitative analysis of the number of cleaved-caspase3-positive cells are shown in lower panel. ( C , D ) Representative Annexin-V/PI images were detected by flow cytometry. Quantitative analysis of the apoptotic rate of SH-SY5Y cells are shown in lower panel. ( E , F ) Primary neuron-glial mixed cells were transfected with S100A8 shRNA vector for 24 h followed by 48 h in hypoxic condition. Cells were harvested, and the expression protein levels of S100A8 and cleaved caspase-3 were analyzed by Western blotting. Data from three independent experiments are presented as the means ± S.D. Values of *** p < 0.001 versus control; # p < 0.05, ### p < 0.001 versus hypoxia-exposed sample were considered as statistically significant.

Article Snippet: S100A8, TNF-α, IL-6, IL-1β and PGE2 were quantitatively measured by an enzyme-linked immunosorbent assay (ELISA) using the human S100A8 Duoset ELISA kits, the mouse TNF-α, IL-6 and IL-1β DuoSet ELISA kits, and the PGE2 parameter assay kit (R&D systems, Minneapolis, MN, USA), according to the manufacturer’s instructions.

Techniques: Expressing, Incubation, Immunofluorescence, Flow Cytometry, Transfection, shRNA, Plasmid Preparation, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Hypoxia-Induced S100A8 Expression Activates Microglial Inflammation and Promotes Neuronal Apoptosis

doi: 10.3390/ijms22031205

Figure Lengend Snippet: The expression of S100A8 in microglial cell induces the Cyclooxygenase-2 (COX-2)/prostaglandin E2 (PGE 2) pathway. BV-2 cells were transfected with S100A8 shRNA or Scramble vector. After 24 h, cells were incubated in hypoxic condition for 48 h. ( A ) The mRNA and ( B ) the protein levels of S100A8 and COX-2 were detected by real-time PCR and western blotting. ( C ) Secretion of PGE 2 level analyzed by ELISA. Data from three independent experiments are presented as the means ± S.D. Values of * p < 0.05, ** p < 0.01 versus control; # p < 0.05, ### p < 0.001 versus hypoxia-exposed sample were considered as statistically significant.

Article Snippet: S100A8, TNF-α, IL-6, IL-1β and PGE2 were quantitatively measured by an enzyme-linked immunosorbent assay (ELISA) using the human S100A8 Duoset ELISA kits, the mouse TNF-α, IL-6 and IL-1β DuoSet ELISA kits, and the PGE2 parameter assay kit (R&D systems, Minneapolis, MN, USA), according to the manufacturer’s instructions.

Techniques: Expressing, Transfection, shRNA, Plasmid Preparation, Incubation, Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Elevated Levels of Neutrophil Activated Proteins, Alpha-Defensins (DEFA1), Calprotectin (S100A8/A9) and Myeloperoxidase (MPO) Are Associated With Disease Severity in COVID-19 Patients

doi: 10.3389/fcimb.2021.751232

Figure Lengend Snippet: Details of the COVID-19 patients examined at presentation.

Article Snippet: Levels of alpha defensins, DEFA1 and S100A8/A9 in serum and plasma were quantified using commercially available ELISA kits, Neutrophil DEFA1 (USCN, Wuhan) & quantikine ELISA for human S100A8/A9 Heterodimer (R & D systems, Minneapolis) as per manufacturer’s protocols.

Techniques:

Journal: Innate immunity

Article Title: CXCL4-induced plaque macrophages can be specifically identified by co-expression of MMP7+S100A8+ in vitro and in vivo.

doi: 10.1177/1753425914526461

Figure Lengend Snippet: Figure 2. MMP7 and S100A8 mRNA expression in CXCL4-induced macrophages. (A, B) Gene expression of S100A8 or MMP7 in monocyte-derived macrophages induced over 6 d with M-CSF (100 ng/ml, M0) or CXCL4 (1 mM, M4) as determined by Affymetrix gene array.17 ***P < 0.001 by local pooled error testing. (C, D) MMP7 or S100A8 gene expression in monocyte-derived macrophages after 6 d in culture with M-CSF (100 ng/ml) or 3 d with M-CSF (100 ng/ml), and additional 3 d with indicated concentrations of CXCL4 as determined by quantitative RT-PCR; n ¼ 3–5, **P < 0.01, *P < 0.05 versus M-CSF. (E, F) MMP7 or S100A8 gene expression in monocyte-derived macrophages after 6 d in culture with M-CSF (100 ng/ml), M-CSF plus heparin (2 U/ml), CXCL4 (1 mM) or CXCL plus heparin (2 U/ml), as determined by quantitative RT-PCR; n ¼ 4–9, **P < 0.01, *P < 0.05 versus CXCL4 plus heparin.

Article Snippet: ELISA kits to measure secretory forms of MMP7 and S100A8 were purchased from Raybiotech (Norcross, at TEXAS SOUTHERN UNIVERSITY on December 17, 2014ini.sagepub.comDownloaded from GA, USA).

Techniques: Expressing, Gene Expression, Derivative Assay, Quantitative RT-PCR

Journal: Innate immunity

Article Title: CXCL4-induced plaque macrophages can be specifically identified by co-expression of MMP7+S100A8+ in vitro and in vivo.

doi: 10.1177/1753425914526461

Figure Lengend Snippet: Figure 3. MMP7 and S100A8 are specific markers for M4 macrophages. (A, B) Radar plots indicating the ratio of S100A8 or MMP7 gene expression in M1-, M2- and M4-polarized macrophages versus unpolarized macrophages derived from the same donor, as determined by Affymetrix gene array.6,17 (C, D) Radar plots indicating the ratio of S100A8 or MMP7 gene expression in M1-, M2- and M4-polarized macrophages versus unpolarized macrophages derived from the same donor as determined by quantitative RT-PCR. (E, F) Radar plots indicating the gene expression of typical marker genes for M1 or M2 macrophage polarization in M1-, M2- and M4-polarized macrophages. In all cases, gene expression was determined by real time RT-PCR and normalized unpolarized macrophages derived from the same donor.

Article Snippet: ELISA kits to measure secretory forms of MMP7 and S100A8 were purchased from Raybiotech (Norcross, at TEXAS SOUTHERN UNIVERSITY on December 17, 2014ini.sagepub.comDownloaded from GA, USA).

Techniques: Gene Expression, Derivative Assay, Quantitative RT-PCR, Marker

Journal: Innate immunity

Article Title: CXCL4-induced plaque macrophages can be specifically identified by co-expression of MMP7+S100A8+ in vitro and in vivo.

doi: 10.1177/1753425914526461

Figure Lengend Snippet: Figure 4. In vitro validation of MMP7, S100A8 as M4 macrophage markers. (A) Representative histograms of intracellular meas- urement of MMP7 protein in monocyte-derived macrophages differentiated with M-CSF or CXCL4. Bar graph indicating relative fluorescence in M-CSF- or CXCL4-induced macrophages, as determined by flow cytometry; n ¼ 6, *P < 0.05 versus M-CSF. (B) Soluble MMP7 in supernatants of monocyte-derived macrophages differentiated with M-CSF (100 ng/ml) or CXCL4 (1 mM) as determined by ELISA; n ¼ 4, P ¼ not significant. (C) Representative images of S100A8 immunostaining in M-CSF- or CXCL4-induced macrophages. (D) Soluble S100A8 in supernatants of monocyte-derived macrophages differentiated with M-CSF (100 ng/ml) or CXCL4 (1 mM) as determined by ELISA; n ¼ 4, *P < 0.05.

Article Snippet: ELISA kits to measure secretory forms of MMP7 and S100A8 were purchased from Raybiotech (Norcross, at TEXAS SOUTHERN UNIVERSITY on December 17, 2014ini.sagepub.comDownloaded from GA, USA).

Techniques: In Vitro, Biomarker Discovery, Derivative Assay, Fluorescence, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Immunostaining

Journal: Innate immunity

Article Title: CXCL4-induced plaque macrophages can be specifically identified by co-expression of MMP7+S100A8+ in vitro and in vivo.

doi: 10.1177/1753425914526461

Figure Lengend Snippet: Figure 5. In vivo evidence for the presence of M4 macrophages in atherosclerotic lesions. Human coronary arteries from patients with coronary artery disease were stained post mortem with (A) hematoxylin and eosin staining or (B) with Abs against CD68 (FITC) and MMP7 (left) or S100A8 (right, both Texas red, magnification 40). (C) Human coronary arteries from patients with coronary artery disease were stained post mortem with Abs against MMP7 (FITC) and S100A8 (Texas red, magnification 40). White boxes are magnified on the right giving examples of (a.) MMP7S100A8+ or (b.) MMP7+S100A8 macrophages. *Vessel lumen.

Article Snippet: ELISA kits to measure secretory forms of MMP7 and S100A8 were purchased from Raybiotech (Norcross, at TEXAS SOUTHERN UNIVERSITY on December 17, 2014ini.sagepub.comDownloaded from GA, USA).

Techniques: In Vivo, Staining

Journal: Frontiers in immunology

Article Title: Extracellular proteins as potential biomarkers in Sepsis-related cerebral injury.

doi: 10.3389/fimmu.2023.1128476

Figure Lengend Snippet: FIGURE 6 Levels of IL-6, CSF3, MMP8, and S100A8 and ROC curves of S100A8 and MMP8 in the peripheral blood of SAE patients. (A) When compared to the non-SAE (n=10) group, the SAE patients (n=21) had higher serum levels of MMP8 and S100A8 (both p<0.0). (B) High diagnostic accuracy was shown for the identification of SAE by both S100A8 (AUC = 0.962, 95% CI = 0.9001-1.000) and MMP8 (AUC = 0.7905, 95% CI = 0.6245-0.9564), with S100A8 having the highest accuracy. (C, D) Correlation analysis of the four extracellular molecules with clinical indicators and prognostic markers. S100A8, MMP8, CSF3, and IL-6 protein levels in the peripheral blood of sepsis patients were significantly linked with GCS scores (all p<0.05). S100A8 levels and 28-day mortality were strongly associated (r=0.634, p<0.01), and IL-6 levels and duration of mechanical breathing were positively correlated (r=0.360, p=0.046). *p <0.05, **p <0.01, ****p <0.0001; ns, no significance.

Article Snippet: The levels of IL6, MMP8, CSF3, and S100A8 were determined by the following commercial ELISA kits, according to manufacturer’s recommendations: MMP8 (Cat.#CSB-E04680h, Cusabio Biotech Co. Ltd, Wuhan, China), S100A8 (Cat. #CSB-E11833h, Cusabio Biotech Co. Ltd, Wuhan, China), IL-6 (Cat. #CSB-E04638h, Cusabio Biotech Co. Ltd, Wuhan, China), and CSF3 (Cat. #EK0360, Boster Biotech Co. Ltd, USA).

Techniques: Diagnostic Assay

Journal: Frontiers in immunology

Article Title: Extracellular proteins as potential biomarkers in Sepsis-related cerebral injury.

doi: 10.3389/fimmu.2023.1128476

Figure Lengend Snippet: FIGURE 7 Levels of MMP8, and S100A8 in the peripheral blood and CSF of sepsis rats. (A)Timeline of experiment procedures. (B) The recognition index of novel items in the CLP model rats (n =11) was lower than that of the Sham group rats (p < 0.05 vs Sham, n = 8). (C) In the Barnes maze experiment, the CLP model rats (n =11) made significantly more mistakes before finding the target hole on postoperative day 9 and 10 compared to the Sham group rats (p < 0.01, p < 0.05 vs Sham, n = 8). (D) The CLP rats (n=14) exhibited significantly elevated serum levels of MMP8 (p<0.05) and CSF levels (p=0.0506) in comparison to the SHAM group (n=12). (E) When compared to the SHAM (n=14) group, the CLP rats (n=12) had higher serum levels of S100A8(p<0.01) and CSF levels (all p<0.0001). CLP, cecal ligation and puncture; NS., no significance; NOR, novel object recognition. *p <0.05; ****p <0.0001.

Article Snippet: The levels of IL6, MMP8, CSF3, and S100A8 were determined by the following commercial ELISA kits, according to manufacturer’s recommendations: MMP8 (Cat.#CSB-E04680h, Cusabio Biotech Co. Ltd, Wuhan, China), S100A8 (Cat. #CSB-E11833h, Cusabio Biotech Co. Ltd, Wuhan, China), IL-6 (Cat. #CSB-E04638h, Cusabio Biotech Co. Ltd, Wuhan, China), and CSF3 (Cat. #EK0360, Boster Biotech Co. Ltd, USA).

Techniques: Comparison, Ligation