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Background: Neutrophil extracellular traps (NETs) are produced by polymorphonuclear neutrophils (PMNs) stimulated by interleukin-17 (IL-17) and tumor necrosis factor α (TNF-α). However, the level and role of NETs in oral lichen planus (OLP) remain poorly understood. Objective: This study aimed to investigate the expression of NETs in OLP and explore the correlation between NETs and the levels of IL-17 and TNF-α. Methods: The expression and distribution of NET-related proteins in tissue samples from each group were assessed using hematoxylin-eosin (HE) staining and immunofluorescence (IF). Additionally, the expression of NET-related proteins in peripheral blood samples from each group was evaluated using cell IF technique and fluorescence spectrophotometry. The relative formation level of NETs in each group was determined by fluorescence spectrophotometry via plasma co-culture. Furthermore, the levels of inflammatory cytokines IL-17 and TNF-α in plasma and culture supernatant were measured using enzyme-linked immunosorbent assay (ELISA). Results: NET-related proteins were located in the subepithelial and lamina propria layers of OLP lesions. OLP had significantly higher expression of NET-related proteins in lesion tissues and peripheral blood compared to the healthy control (HC) group (p < 0.05). The rate of NETs formation in the erosive-stage OLP (EOLP) group was significantly higher than that in the HC group (p < 0.05), in contrast, no significant increase was observed in the non-erosive OLP (NEOLP) group (p > 0.05). Furthermore, the levels of IL-17 and TNF-α in the EOLP group were significantly elevated compared to those in the NEOLP group and HC group (p < 0.05), while the levels in the NEOLP group did not significantly differ from those in the HC group (p > 0.05). The rate of NETs formation showed a positive correlation with the levels of IL-17 and TNF-α in plasma. Conclusion: The expression of NET-related proteins was upregulated in OLP lesion tissues and peripheral blood. Elevated levels of IL-17 and TNF-α in peripheral blood plasma positively correlated with the rate of NETs formation, suggesting that IL-17 and TNF-α mediate the formation of NETs in OLP patients, and may thereby contribute to the development of OLP. [ABSTRACT FROM AUTHOR]

Background. Chronic endometritis (CE) is an inflammatory hysteropathy causing miscarriage and infertility. High invasiveness of the main method of CE diagnosis and vague clinical picture necessitate the need for the development of less invasive approaches to establish the presence of this disease.The aim of the study. To establish a significant association of the concentration of pro- and anti-inflammatory interleukins in the blood serum with the presence of chronic endometritis in premenopausal women without concomitant endocrine diseases.Materials and methods. This re-analysis of the data is based on the results of a cross-sectional study conducted between May 2017 and December 2019 which included 198 premenopausal women. In all participants, body weight and height were measured with the calculation of the body mass index, the concentration of Creactive protein, pro- and anti-inflammatory cytokines in the blood serum was determined, and a pipelle biopsy was performed to determine the CD138 expression in the endometrial stroma. Non-parametric methods, as well as ROC analysis, were used for statistical analysis.Results. 86 women were included in the re-analysis of the data, 37 of them had a confirmed diagnosis of chronic endometritis. Statistically significantly higher values of interleukin 1 (IL-1) concentration (p = 0.0028) and IL-1/tumor necrosis factor α ratio (p < 0.001) were determined in women with CE and normal body weight; threshold values of these parameters were ≥ 1.35 pg/ml (sensitivity 75 %, specificity 83 %; 95% confidence interval (95% CI): 0.88–2.15) and ≥ 1.03 (sensitivity 85 %, specificity 78 %; 95% CI: 0.81–1.27) respectively. Such a relationship was not revealed in women with overweight.Conclusions. The obtained results can be the basis for conducting a larger-scale study with determining the concentration of cytokines not only in the blood serum, but also in the endometrium of women with CE, which will allow the development of a minimally invasive method for determining the risk of the presence of chronic endometritis in premenopausal women.

Thrombosis is a key process that determines acute coronary syndrome and ischemic stroke and is the leading cause of morbidity and mortality in the world, together with cancer. Platelet adhesion and subsequent activation and aggregation are critical processes that cause thrombus formation after endothelial damage. To date, high hopes are associated with compounds of natural origin, which show anticoagulant action without undesirable effects and can be proposed as supportive therapies. We investigated the effect of the new combination of four natural compounds, escin–bromelain–ginkgo biloba–sage miltiorrhiza (EBGS), on the initial process of the coagulation cascade, which is the adhesion of platelets to activated vascular endothelium. Our results demonstrated that EBGS pretreatment of endothelial cells reduces platelet adhesion even in the presence of the monocyte–lymphocyte population. Our data indicate that EBGS exerts its effects by inhibiting the transcription of adhesion molecules, including P-selectin, platelet membrane glycoprotein GP1b, integrins αV and β3, and reducing the secretion of the pro-inflammatory cytokines interleukin 6, interleukin 8, and the metalloproteinases MMP-2 and MMP-9. Furthermore, we demonstrated that EBGS inhibited the expression of focal adhesion kinase (FAK), strictly involved in platelet adhesion, and whose activity is correlated with that of integrin β3. The results shown in this manuscript suggest a possible inhibitory role of the new combination EBGS in the reduction in platelet adhesion to activated endothelium, thus possibly preventing coagulation cascade initiation. [ABSTRACT FROM AUTHOR]

Background Neutrophil extracellular traps (NETs) are produced by polymorphonuclear neutrophils (PMNs) stimulated by interleukin-17 (IL-17) and tumor necrosis factor α (TNF-α). However, the level and role of NETs in oral lichen planus (OLP) remain poorly understood. Objective This study aimed to investigate the expression of NETs in OLP and explore the correlation between NETs and the levels of IL-17 and TNF-α. Methods The expression and distribution of NET-related proteins in tissue samples from each group were assessed using hematoxylin-eosin (HE) staining and immunofluorescence (IF). Additionally, the expression of NET-related proteins in peripheral blood samples from each group was evaluated using cell IF technique and fluorescence spectrophotometry. The relative formation level of NETs in each group was determined by fluorescence spectrophotometry via plasma co-culture. Furthermore, the levels of inflammatory cytokines IL-17 and TNF-α in plasma and culture supernatant were measured using enzyme-linked immunosorbent assay (ELISA). Results NET-related proteins were located in the subepithelial and lamina propria layers of OLP lesions. OLP had significantly higher expression of NET-related proteins in lesion tissues and peripheral blood compared to the healthy control (HC) group (p < 0.05). The rate of NETs formation in the erosive-stage OLP (EOLP) group was significantly higher than that in the HC group (p < 0.05), in contrast, no significant increase was observed in the non-erosive OLP (NEOLP) group (p > 0.05). Furthermore, the levels of IL-17 and TNF-α in the EOLP group were significantly elevated compared to those in the NEOLP group and HC group (p < 0.05), while the levels in the NEOLP group did not significantly differ from those in the HC group (p > 0.05). The rate of NETs formation showed a positive correlation with the levels of IL-17 and TNF-α in plasma. Conclusion The expression of NET-related proteins was upregulated in OLP lesion tissues and peripheral blood. Elevated levels of IL-17 and TNF-α in peripheral blood plasma positively correlated with the rate of NETs formation, suggesting that IL-17 and TNF-α mediate the formation of NETs in OLP patients, and may thereby contribute to the development of OLP.

Numerous evidences showed that human umbilical cord blood (UCB) mononuclear cells were a promising approach for the therapy of ischemic stroke(IS). The effect of stage-specific embryonic antigen 3 (SSEA3）positive subpopulation in UCB was not investigated in IS. In this study, we isolated SSEA3 positive cells from healthy UCB mononuclear cells, which comprised about 7.01% of the total UCB-mononuclear cells. Flow cytometry analysis revealed that SSEA3(+)UCB cells were almost positive for CD44 and CD45, and negative for CD73, CD90 and CD105. The expression of Oct3/4 in SSEA3(+)UCB cells were higher than that in UCB. SSEA3(+)UCB cells sorted by magnetic cell sorting were intravenously injected into the middle cerebral arterial occlusion(MCAO) rat model. Neurological score showed that SSEA3(+)UCB transplantation group exhibited significant improvements in the functional outcome of MCAO rats than UCB transplantation group. Nissl staining and microtubule association protein-2(MAP2) immunofluorescence staining showed that the SSEA3(+)UCB transplantation group decreased neuronal loss. SSEA3(+)UCB transplantation group reduced neuronal apoptosis, inhibited caspase3 expression, and decreased tumor necrosis factor α(TNF-α). These results indicate that SSEA3 positive cells are a novel subpopulation of UCB cells, which exhibit great potential for the treatment of ischemic stroke.

Severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) induced cytokine storm is the major cause of COVID-19 related deaths. Patients have been treated with drugs that work by inhibiting a specific protein partly responsible for the cytokines production. This approach provided very limited success, since there are multiple proteins involved in the complex cell signaling disease mechanisms. We targeted five proteins: Angiotensin II receptor type 1 (AT1R), A disintegrin and metalloprotease 17 (ADAM17), Nuclear Factor‑Kappa B (NF‑κB), Janus kinase 1 (JAK1) and Signal Transducer and Activator of Transcription 3 (STAT3), which are involved in the SARS‑CoV‑2 induced cytokine storm pathway. We developed machine-learning (ML) models for these five proteins, using known active inhibitors. After developing the model for each of these proteins, FDA-approved drugs were screened to find novel therapeutics for COVID‑19. We identified twenty drugs that are active for four proteins with predicted scores greater than 0.8 and eight drugs active for all five proteins with predicted scores over 0.85. Mitomycin C is the most active drug across all five proteins with an average prediction score of 0.886. For further validation of these results, we used the PyRx software to conduct protein-ligand docking experiments and calculated the binding affinity. The docking results support findings by the ML model. This research study predicted that several drugs can target multiple proteins simultaneously in cytokine storm-related pathway. These may be useful drugs to treat patients because these therapies can fight cytokine storm caused by the virus at multiple points of inhibition, leading to synergistically effective treatments.

Numerous evidences showed that human umbilical cord blood (UCB) mononuclear cells were a promising approach for the therapy of ischemic stroke(IS). The effect of stage-specific embryonic antigen 3 (SSEA3）positive subpopulation in UCB was not investigated in IS. In this study, we isolated SSEA3 positive cells from healthy UCB mononuclear cells, which comprised about 7.01% of the total UCB-mononuclear cells. Flow cytometry analysis revealed that SSEA3(+)UCB cells were almost positive for CD44 and CD45, and negative for CD73, CD90 and CD105. The expression of Oct3/4 in SSEA3(+)UCB cells were higher than that in UCB. SSEA3(+)UCB cells sorted by magnetic cell sorting were intravenously injected into the middle cerebral arterial occlusion(MCAO) rat model. Neurological score showed that SSEA3(+)UCB transplantation group exhibited significant improvements in the functional outcome of MCAO rats than UCB transplantation group. Nissl staining and microtubule association protein-2(MAP2) immunofluorescence staining showed that the SSEA3(+)UCB transplantation group decreased neuronal loss. SSEA3(+)UCB transplantation group reduced neuronal apoptosis, inhibited caspase3 expression, and decreased tumor necrosis factor α(TNF-α). These results indicate that SSEA3 positive cells are a novel subpopulation of UCB cells, which exhibit great potential for the treatment of ischemic stroke. • SSEA3 positive subpopulation in umbilical cord blood were positive for CD44 and CD45. • SSEA3 + UCB alleviated cerebral infarction after ischemic stroke. • SSEA3 + UCB treatment decreased cell apoptosis in brain infarction. [ABSTRACT FROM AUTHOR]

Introduction: Uterine fibroids (UFs) are benign tumors of the female reproductive system originating from the smooth muscle of the uterus. Currently, progesterone is known to play a key role in the differentiation of the myometrial tissue to form UFs and their abnormal growth. The mechanism of action of progesterone in UF tumorigenesis involves its effect on increasing the concentrations and dysregulation of selected growth factors. Material and methods: A retrospective cohort study was performed to evaluate and compare tumor necrosis factor a (TNF-α), insulin-like growth factor 1 (IGF-1), plasminogen activator inhibitor-1 (PAI-1) serum concentrations in patients with UFs without prior hormonal treatment, patients with UFs treated with a 3-month standard ulipristal acetate (UPA - a type of selective progesterone receptor modulator) scheme (5 mg/day) and in control patients without UFs. A total of 120 patients were divided into 3 groups (controls, UFs with UPA treatment, UFs without UPA treatment). Results: There were no significant differences in TNF-α serum concentrations between patients with UFs who underwent UPA treatment and patients who did not. Serum concentrations of IGF-1 and PAI-1 did not show significant intergroup differences. Conclusions: No significant differences were found between TNF-α concentrations in the serum of patients with UFs treated with UPA, and patients without UPA treatment. In addition, our data analysis did not show significant differences in the concentrations of IGF-1 and PAI-1 between patients with UFs and the control group. Further studies on the dependence of specific symptoms on selected growth factors are mandatory. [ABSTRACT FROM AUTHOR]

BACKGROUND: Intervertebral disc degeneration is an important cause of low back pain. At present, there are many modeling methods for disc degeneration in China and abroad, but there is not a model for low back pain due to disc degeneration. OBJECTIVE: To compare the effect of mechanical puncture combined with tumor necrosis factor α and complete Freund’s adjuvant with a conventional disc mechanical puncture alone. METHODS: A total of 18 male adult Sprague-Dawley rats were randomly divided into 3 groups, with 6 animals in each group. No treatment was given in the blank group. Animal models of intervertebral disc degeneration were made in the L4-5 segments of rats in the control using conventional mechanical puncture. In the experimental group, on the basis of mechanical puncture, tumor necrosis factor α+complete Freund’s adjuvant was injected into the L4-5 intervertebral discs using a microinjector to establish a model of disc degeneration induced by mechanical puncture combined with inflammatory factors. Four weeks after surgery, the pain threshold of rats was measured by the hot plate method for assessing the perception of heat injury in rats with intervertebral disc degeneration. MRI examination was performed to observe the disc degeneration in each group. ELISA was used to detect the levels of serum tumor necrosis factor α, interleukin 1β, interleukin 6 and prostaglandin E2. Hematoxylin-eosin and Safranin O-fast green staining were used to observe the morphological changes of the disc. RESULTS AND CONCLUSION: In terms of pain, the behavioral pain threshold of the experimental group was continuously decreased, and the levels of serum inflammatory factors were significantly higher compared with the control group. In terms of morphology, the MRI results showed that the L4-5 nucleus pulposus signal completely disappeared in the experimental group. Histopathological results showed that in the control group, the nucleus pulposus was intact, more notochord cells were visible, and some fiber rings were ruptured, while in the experimental group, there are fewer notochord cells and the structure of the nucleus pulposus and fibrous ring is disturbed, with the boundary disappearing. To conclude, mechanical puncture combined with tumor necrosis factor alpha and complete Freund’s adjuvant can successfully establish a discogenic low back pain model in rats. This operation is simple and economical to achieve obvious disc degeneration and low back pain, with greatly shortened molding cycle. This model can be used as a reference for studying discogenic low back pain models. [ABSTRACT FROM AUTHOR]

Pulmonary thromboembolism caused by thrombi blocking major pulmonary artery and its branches, is a frequently encountered phenomenon and an important cause of high morbidity and mortality in lung diseases and may develop into persistent pulmonary hypertension (PH). Nuclear factor‐κB (NF‐κB) signaling pathway had been reported participated in the formation and development of PH by promoting inflammatory response. The aim of this study was to investigate the effects of NF‐κB activation on the serum levels of tumor necrosis factor α (TNF‐α) and interleukin‐1β (IL‐1β) in acute pulmonary microthromboembolism (APMTE) rats. Rats were randomized into five groups. APMTE group received jugular vein injection of autologous thrombus, while control group rats received normal saline injection. Pulmonary hemodynamic parameters were measured through ECHO‐guided transthoracic puncture. Pulmonary vascular morphological changes were analyzed by HE. The expression changes of NF‐κB and serum TNF‐α、IL‐1β levels were detected by enzyme‐linked immunosorbent assay. Protein expression of the MAPK/NF‐κB signaling pathway including p‐IκBα, p‐p38 MAPK, p‐NF‐κB p65, IκBα, p38 MAPK, and NF‐κB p65 was determined using western blot analysis. Compared with control group, the expression of NF‐κB in lung tissue and the levels of serum TNF‐α and IL‐1β rats were higher, a significant reduction in IκBα and elevation in the phosphorylation of IκBα, p38 MAPK, and NF‐κB p65 were found in APMTE group rats. And UK administration reversed the APMTE‐induced increase in TNF‐α, IL‐1β, p‐IκBα, p‐MAPK, and p‐NF‐κB protein. Furthermore, the levels of NF‐κB, TNF‐α, and IL‐1β were positively correlated with mean pulmonary artery. And the levels of TNF‐α and IL‐1β were positively correlated with NF‐κB. These findings suggest that the activation of MAPK/NF‐κB pathway as a critical driver of increasing TNF‐α and IL‐1β level in APMTE rats and UK exerted protective effects against APMTE‐induced PH may be related to the downregulation of the MAPK/NF‐κB signaling pathway. [ABSTRACT FROM AUTHOR]

Purpose: At present, therapeutic interventions to treat acute lung injury (ALI) remain largely limited to lung-protective strategies, as no real molecular-driven therapeutic intervention has yet become available. The administration of bacterial lipopolysaccharides (LPS) is known as an inflammatory activator, representing a frequently used model of ALI. This study investigated the biological function of normoxic (21% O2) vs. hypoxic conditions (5% O2) obtained from human Wharton's Jelly mesenchymal stem cells (hWJ-MSCs) and discovered that exosomes have the ability to suppress inflammatory responses by specifically targeting TNF-α, IL-1β, IL-6. and identify the toll-like receptor 4 (TLR4) NF-κβ gene expression. Methods: Primer culture hWJ-MSCs characterization with trilineage differentiation and CD markers was conducted. To obtain exosomes, hWJ-MSCs were stimulated with two different oxygen levels: 21% (nor-exo) and 5% (hypo-exo). Then, the L2 cell line was induced with LPS 1 µg/mL. Inflamed-L2 was treated with nor-exo, hypo-exo, and dexamethasone as a positive control. The RNA extracted from treated L2 cells was utilized to examine the gene expression profiles of TLR4 and NF-κβ, and the medium was used to measure tumor necrosis factor α (TNF-α), interleukin (IL)-1β, and IL-6 levels using ELISA. Lastly, proteomic analysis of the exosome using LC/MS-MS was conducted. Results: Nor-exo and hypo-exo can be characterized and can produce higher yields exosomes under hypoxic conditions. The expression of TLR4 and NF-κβ genes and the proinflammatory levels such as IL-6, IL-1β, and TNF-α levels in nor-exo and hypo-exo treatments decreased. Conclusion: Nor-exo and hypo-exo derived from hWJ-MSCs were proven to have anti-inflammatory activities. [ABSTRACT FROM AUTHOR]

The purpose of this study was to compare the inflammatory biomarkers in postmenopausal women with osteoporosis and those with normal bone mineral density (BMD).A total of 850 postmenopausal women aged 50 to 65 were randomly selected for participation in this cross-sectional investigation. 100 women displayed normal BMD, while 101 were diagnosed with osteoporosis, as determined by dual-energy X-ray absorptiometry. Biochemical techniques were used to quantify tumor necrosis factor α (TNF-α) levels, high-sensitivity C-reactive protein (hs-CRP), and interleukin-6. The area under the curve (AUC) for the diagnosis of osteoporosis was calculated using receiver-operator characteristic (ROC) curves.A significant difference was observed between the two groups in terms of age, menopause age, education level, and BMI (p

Introduction: Alzheimerʼs disease (AD) has been identified as a progressive memory and cognitive impairment. Some Salvia species are suggested by certain studies for the management of mild to moderate AD. We aimed to evaluate the anti-inflammatory, antioxidant, and anti-apoptotic effects of S. hydrangea on amyloid beta-injected rats. Materials and Methods: Rats were pretreated with S. hydrangea for 10 days before amyloid beta (Aβ) injection. Western blotting techniques were used to evaluate protein level of γ-glutamyl cysteine synthetase (γ-GCS), Tumor Necrosis Factor α (TNF-α) and Interleukin-6 (IL-6) in two brain regions: hippocampus and frontal cortex. Results: Current data show that S. hydrangea extract increased γ-GCS protein levels in amyloid beta injected rats, and pretreatment with S. hydrangea increased it further. Besides, S. hydrangea decreased protein levels of TNF-α and IL-6 in amyloid beta injected rats. Conclusion: Based on the decreased levels of IL-6, TNF-α, and the increased levels of γ-GCS, it is suggested that the use of S. hydrangea could be protective in neurodegenerative diseases.

Background: To study the correlation between the expression of immune factors and the prognosis of surgical treatment of plasma cell mastitis (PCM) and to analyze its predictive value for the prognosis of patients. Methods: 89 female patients with PCM treated in our hospital from June 2020 to September 2022 were divided into good prognosis group (69 cases) and poor prognosis group (20 cases). Logistic regression was used to analyze the influencing factors of poor prognosis of surgical treatment for PCM, and to explore the correlation between these immune factors and the prognosis of surgical treatment for PCM. Draw the receiver operating characteristic (ROC) curve to analyze the predictive value of the above indexes for the prognosis of patients with PCM. Results: After 4 weeks of treatment, the levels of tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) were significantly lower than before treatment (p < 0.05), and the level of interleukin 10 (IL-10) was significantly higher than before treatment (p < 0.05). At the time of admission, there was no significant difference in the clinical data and the levels of TNF-α, IL-6 and IL-10 between the two groups (p > 0.05). After 4 weeks of treatment, the indexes of TNF-α and IL-6 in the poor prognosis group were higher than those in the good prognosis group (p < 0.05). Logistic regression analysis showed that the levels of TNF-α (odds ratio (OR) = 1.551, 95% confidence interval (95% CI): 1.276–1.886) and IL-6 (OR = 1.082, 95% CI: 1.046–1.119) were increased, which were risk factors for the prognosis of PCM (OR >1). Correlation analysis showed that TNF-α and IL-6 were negatively correlated with the prognosis of PCM, while IL-10 was positively correlated with the prognosis of PCM. ROC curve analysis showed that the areas under the curve for TNF-α, IL-6 and IL-10 to predict the prognosis of surgical treatment of PCM were 0.896, 0.931 and 0.709 respectively. Conclusions: The expression of immune factors such as TNF-α, IL-6 and IL-10 is closely related to the prognosis of surgical treatment of PCM, which has high predictive value for its prognosis.

The current gold standard to treat large cartilage defects is autologous chondrocyte transplantation (ACT). As a new surgical method of cartilage regeneration, minced cartilage implantation (MCI) is increasingly coming into focus. The aim of this study is to investigate the influence of chondrogenesis between isolated and cultured chondrocytes compared to cartilage chips in a standardized inflammation model with the proinflammatory cytokine TNFα. Articular chondrocytes from bovine cartilage were cultured according to the ACT method to passage 3 and transferred to spheroid culture. At the same time, cartilage was fragmented (<1 mm3) to produce cartilage chips. TNFα (20 ng/mL) was supplemented to simulate an inflammatory process. TNFα had a stronger influence on the passaged chondrocytes compared to the non-passaged ones, affecting gene expression profiles differently between isolated chondrocytes and cartilage chips. MCI showed less susceptibility to TNFα, with reduced IL-6 release and less impact on inflammation markers. Biochemical and histological analyses supported these findings, showing a greater negative influence of TNFα on the passaged pellet cultures compared to the unpassaged cells and MCI constructs. This study demonstrated the negative influence of TNFα on chondrogenesis in a chondrocyte spheroid culture and cartilage fragment model. Passaged chondrocytes are more sensitive to cytokine influences compared to non-passaged cells and chondrons. This suggests that MCI may have superior regeneration potential in osteoarthritic conditions compared to ACT. Further investigations are necessary for the translation of these findings into clinical practice. [ABSTRACT FROM AUTHOR]

Background: Our previous studies demonstrated that sour cherry anthocyanins (AC) reduce the salivary count of Streptococcus mutans and inhibit salivary amylase activity within 30 minutes after chewing AC gum. AC gum and changing toothbrushes after scaling reduced the Gram-negative species in the unstimulated salivary microbiota. The present study examined the effect of AC gums on salivary factors, including changes in microbiome. Methods: The study was conducted over three weeks with two groups; young adults (18–30) and adults (30–45). Ten participants changed their toothbrushes, while the other 10 participants did not change after the control period. After scaling, all participants received three doses of AC gum daily. The salivary mRNA and protein levels of cytokines, mucins, melatonin, and the microbiota of unstimulated and stimulated saliva were determined by polymerase chain reaction, enzyme-linked immunosorbent assay, and 16S rRNA gene sequencing. Results: Significantly higher levels of tumor necrosis factor α (TNFα), interleukin-1β (IL-1β), mucin5B (MUC5B), mucin7 (MUC7), and melatonin were detected in stimulated saliva. Correlation analysis of these factors with the microbiota showed positive correlations with the genera Lachnospiraceae, Eikenella, Saccharibacteria_(TM7), Streptococcus, Prevotella, and Haemophilus. Conclusions: AC chewing gum has a beneficial effect on the composition of the oral microbiome, and toothbrush replacement leads to changes in the levels of salivary pro-inflammatory cytokines. [ABSTRACT FROM AUTHOR]

Background: Recent epidemiological studies suggested correlation between gastric cancer (GC) and periodontal disease. Aims: We aim to clarify involvement of lipopolysaccharide of Porphyromonas gingivalis (Pg.), one of the red complex periodontal pathogens, in the GC development. Methods: To evaluate barrier function of background mucosa against the stimulations, we applied biopsy samples from 76 patients with GC using a Ussing chamber system (UCs). K19-Wnt1/C2mE transgenic (Gan) mice and human GC cell-lines ± THP1-derived macrophage was applied to investigate the role of Pg. lipopolysaccharide in inflammation-associated carcinogenesis. Results: In the UCs, Pg. lipopolysaccharide reduced the impedance of metaplastic and inflamed mucosa with increases in mRNA expression of toll-like receptor (TLR) 2, tumor necrosis factor (TNF) α, and apoptotic markers. In vitro, Pg. lipopolysaccharide promoted reactive oxidative stress (ROS)-related apoptosis as well as activated TLR2-β-catenin-signaling on MKN7, and it increased the TNFα production on macrophages, respectively. TNFα alone activated TLR2-β-catenin-signaling in MKN7, while it further increased ROS and TNFα in macrophages. Under coculture with macrophages isolated after stimulation with Pg. lipopolysaccharide, β-catenin-signaling in MKN7 was activated with an increase in supernatant TNFα concentration, both of which were decreased by adding a TNFα neutralization antibody into the supernatant. In Gan mice with 15-week oral administration of Pg. lipopolysaccharide, tumor enlargement with β-catenin-signaling activation were observed with an increase in TNFα with macrophage infiltration. Conclusions: Local exposure of Pg. lipopolysaccharide may increase ROS on premalignant gastric mucosa to induce apoptosis-associated barrier dysfunction and to secrete TNFα from activated macrophages, and both stimulation of Pg. lipopolysaccharide and TNFα might activate TLR2-β-catenin-signaling in GC. [ABSTRACT FROM AUTHOR]

Abstract Pulmonary thromboembolism caused by thrombi blocking major pulmonary artery and its branches, is a frequently encountered phenomenon and an important cause of high morbidity and mortality in lung diseases and may develop into persistent pulmonary hypertension (PH). Nuclear factor‐κB (NF‐κB) signaling pathway had been reported participated in the formation and development of PH by promoting inflammatory response. The aim of this study was to investigate the effects of NF‐κB activation on the serum levels of tumor necrosis factor α (TNF‐α) and interleukin‐1β (IL‐1β) in acute pulmonary microthromboembolism (APMTE) rats. Rats were randomized into five groups. APMTE group received jugular vein injection of autologous thrombus, while control group rats received normal saline injection. Pulmonary hemodynamic parameters were measured through ECHO‐guided transthoracic puncture. Pulmonary vascular morphological changes were analyzed by HE. The expression changes of NF‐κB and serum TNF‐α、IL‐1β levels were detected by enzyme‐linked immunosorbent assay. Protein expression of the MAPK/NF‐κB signaling pathway including p‐IκBα, p‐p38 MAPK, p‐NF‐κB p65, IκBα, p38 MAPK, and NF‐κB p65 was determined using western blot analysis. Compared with control group, the expression of NF‐κB in lung tissue and the levels of serum TNF‐α and IL‐1β rats were higher, a significant reduction in IκBα and elevation in the phosphorylation of IκBα, p38 MAPK, and NF‐κB p65 were found in APMTE group rats. And UK administration reversed the APMTE‐induced increase in TNF‐α, IL‐1β, p‐IκBα, p‐MAPK, and p‐NF‐κB protein. Furthermore, the levels of NF‐κB, TNF‐α, and IL‐1β were positively correlated with mean pulmonary artery. And the levels of TNF‐α and IL‐1β were positively correlated with NF‐κB. These findings suggest that the activation of MAPK/NF‐κB pathway as a critical driver of increasing TNF‐α and IL‐1β level in APMTE rats and UK exerted protective effects against APMTE‐induced PH may be related to the downregulation of the MAPK/NF‐κB signaling pathway.

BACKGROUND: Previous studies had shown that hair follicle epidermal neural crest stem cells have the great potential in facilitating peripheral nerve rehabilitation, but the role in regulating inflammation after facial nerve injury had rarely been reported. OBJECTIVE: To investigate whether epidermal neural crest stem cells can modify the expression of pro-inflammatory factor tumor necrosis factor α and anti-inflammatory factor interleukin-4 during facial nerve regeneration to promote the functional and morphological recovery of the facial nerve. METHODS: Epidermal neural crest stem cells were extracted from one 4-day-old SD rat, cultured and identified. Fifty-four adult SD rats were used to construct the bridge model of facial nerve trunk defect with the autologous venous catheter. Models were randomly and equally divided into the normal saline group, Dulbecco's modified eagle medium group and epidermal neural crest stem cell group. Tumor necrosis factor α and interleukin-4 expression levels were detected at postoperative 4 to 14 days using western blotting, immunohistochemistry, and immunofluorescence. The facial nerve function of rats was scored 12 weeks after the operation and the morphology of the facial nerve was observed by hematoxylin-eosin staining. RESULTS AND CONCLUSION: (1) Double positive expression of Nestin and p75NTR could be observed in epidermal neural crest stem cells and the cell purity was over 90%. (2) Compared with the other two groups, tumor necrosis factor α expression in the epidermal neural crest stem cell group turned weaker 7 days after surgery (P < 0.05), and interleukin-4 expression in the epidermal neural crest stem cell group was enhanced on postoperative 3, 7 and 14 days, respectively (P < 0.05). (3) The recovery score of facial nerve function in the epidermal neural crest stem cell group was significantly better than that in the other two groups (P < 0.05). (4) There were abundant nutrient vessels and regenerative axons in the graft segment. Compared with the other two groups, the arrangement of nerve fibers was more orderly and the wrapped myelin sheath was thicker in the graft and distal segment in the epidermal neural crest stem cell group. (5) The data inferred that epidermal neural crest stem cells regulate the local expression of tumor necrosis factor α and interleukin-4 at the early stage of facial nerve injury, inhibit the local inflammatory reaction, and improve facial nerve function and morphology of the regenerated nerve. [ABSTRACT FROM AUTHOR]

ABSTRACT: Heat stress negatively affects the metabolism and physiology of the bovine gut. However, it is not known whether heat stress induces an inflammatory response in mesenteric lymph nodes (MLN), the primary origin of gut immune cells, and thus contributes to inflammatory processes in the circulation. Therefore, our objective was to elucidate the effects of chronic heat stress on the systemic activation of acute-phase response in blood, proinflammatory cytokine production in peripheral blood mononuclear cells (PBMC), and the activation of the toll-like receptor signaling (TLR) 2/4 pathway in MLN leucocytes and their chemokines and chemokine receptor profiles in Holstein cows. Primiparous Holstein cows (n = 30; 169 ± 9 d in milk) were exposed to a temperature-humidity index (THI) of 60 [16°C, 63% relative humidity (RH)] for 6 d. Thereafter, cows were evenly assigned to 3 groups: heat-stressed (HS; 28°C, 50% RH, THI = 76), control (CON; 16°C, 69% RH, THI = 60), or pair-feeding (PF; 16°C, 69% RH, THI = 60) for 7 d. On d 6, PBMC were isolated and on d 7 MLN. Plasma haptoglobin, TNFα, and IFNγ concentrations increased more in HS than CON cows. Concomitantly, TNFA mRNA abundance was higher in PBMC and MLN leucocytes of HS than PF cows, whereas IFNG mRNA abundance tended to be higher in MLN leucocytes of HS than PF cows, but not for chemokines (CCL20, CCL25) or chemokine receptors (ITGB7, CCR6, CCR7, CCR9). Furthermore, the TLR2 protein expression tended to be more abundant in MLN leucocytes of HS than PF cows. These results suggest that heat stress induced an adaptive immune response in blood, PBMC, and MLN leukocytes involving the acute-phase protein haptoglobin, proinflammatory cytokine production, and TLR2 signaling in MLN leucocytes. However, chemokines regulating the leucocyte trafficking between MLN and gut seem not to be involved in the adaptive immune response to heat stress.

Hypoxia can induce pulmonary edema (PE) and inflammation. Furthermore, hypoxia depresses left ventricular (LV) inotropy despite sympathetic activation. To study the role of hypoxic sympathetic activation, we investigated the effects of hypoxia with and without adrenergic blockade (AB) on cardiovascular dysfunction and lung injury, i.e., pulmonary edema, congestion, inflammation, and nitrosative stress. Eighty-six female rats were exposed for 72 h to normoxia or normobaric hypoxia and received infusions with NaCl, prazosin, propranolol, or prazosin–propranolol combination. We evaluated hemodynamic function and performed histological and immunohistochemical analyses of the lung. Hypoxia significantly depressed LV but not right ventricular (RV) inotropic and lusitropic functions. AB significantly decreased LV function in both normoxia and hypoxia. AB effects on RV were weaker. Hypoxic rats showed signs of moderate PE and inflammation. This was accompanied by elevated levels of tumor necrosis factor α (TNFα) and nitrotyrosine, a marker of nitrosative stress in the lungs. In hypoxia, all types of AB markedly reduced both TNFα and nitrotyrosine. However, AB did not attenuate PE. The results suggest that hypoxia-induced sympathetic activation contributes to inflammation and nitrosative stress in the lungs but not to PE. We suggest that AB in hypoxia aggravates hypoxia-induced inotropic LV dysfunction and backlog into the pulmonary circulation, thus promoting PE.

Introduction: Multiple sclerosis (MS) is the most common demyelinating disease of the central nervous system. However, the limitations of available therapeutic strategies are frustrating, both in terms of their low efficacy and multiple side effects. Previous studies showed that natural compounds such as Chalcones possess neuroprotective effects on neurodegenerative disorders. However, few studies have so far been published on the potential effects of Chalcones on treating demyelinating disease. The present study was designed to investigate the effects of Chalcones from Ashitaba (ChA) on cuprizone‐induced noxious changes in the C57BL6 mice model of MS. Methods: The mice received normal diets (Control group: CNT), or Cuprizone‐supplemented diets either without ChA (Cuprizone group: CPZ) or with low or high (300, 600 mg/kg/day) doses of ChA (ChA‐treated groups: CPZ+ChA300/600). Brain‐derived neurotrophic factor (BDNF) and tumor necrosis factor alpha (TNFα) levels, demyelination scores in the corpus callosum (CC), and cognitive impairment were evaluated using the enzyme‐linked immunosorbent assay, histological, and Y‐maze tests, respectively. Results: The findings showed that ChA Co‐treatment significantly reduced the extent of demyelination in the CC and the serum and brain levels of TNFα in the ChA‐treated groups compared to the CPZ group. Besides, treatment with a higher dose of ChA significantly improved the behavioral responses and BDNF levels in the serum and brain of the CPZ+ChA600 group when compared with the CPZ group. Conclusion: The present study provided evidence for the neuroprotective effects of ChA on cuprizone‐induced demyelination and behavioral dysfunction in C57BL/6 mice, possibly by modulating TNFα secretion and BDNF expression. [ABSTRACT FROM AUTHOR]

Nowadays energy drinks (EDs) are widely used among teenagers. One of the most popular EDs in Egypt is Red BullR (RB). The aim of this study was to evaluate the chronic effects of RB on gastric acidity, oxidative stress, and inflammation in pylorus-ligated rats. Thirty-two adult male albino rats were divided into 4 groups; Control animals were given distilled water instead of a beverage. RB was given in doses of 5, 7.5, and 10 ml/kg, p.o., twice daily for 28 days in pylorus-ligated rats. The effects of different doses of RB were investigated on body weight, gastric acidity, and volume and pH of gastric secretions. Biochemical indices of oxidative status including malondialdehyde (MDA), glutathione (GSH), and nitric oxide (NO) were also measured. In addition, interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α) were evaluated. Pretreatment with RB significantly increased body weight, gastric juice volume, acidity, and decreased pH in a dose-dependent manner. Moreover, oral administration of RB resulted in a significant increase in levels of MDA, NO, IL-6, and TNF-α along with decreasing GSH in stomach homogenate. The stomach of rats treated with RB showed sporadic focal records of apoptotic bodies in lining mucosal cells. Moreover; higher records of mucosal/submucosal inflammatory cell infiltrate were observed with more severe submucosal edema. Chronic consumption of RB increased gastric acidity, oxidative stress, and inflammation along with decreasing mucus secretion in rat stomach. [ABSTRACT FROM AUTHOR]

Adipose-derived mesenchymal stem cells are increasingly being used in regenerative medicine as cell therapy targets, including in the treatment of burns and ulcers. The regenerative potential of AD-MSCs and some of their immunological properties are known from in vitro studies; however, in clinical applications, cells are used in non-ideal conditions and can behave differently in inflammatory environments, affecting the efficacy and outcome of therapy. Our aim was to investigate and map the pathways that the inflammatory microenvironment can induce in these cells. High-throughput gene expression assays were performed on AD-MSCs activated with LPS and TNFα. Analysis of RNA-Seq data showed that control, LPS-treated and TNFα-treated samples exhibited distinct gene expression patterns. LPS treatment increased the expression of 926 genes and decreased the expression of 770 genes involved in cell division, DNA repair, the cell cycle, and several metabolic processes. TNFα treatment increased the expression of 174 genes and decreased the expression of 383 genes, which are related to cell division, the immune response, cell proliferation, and differentiation. We also map the biological pathways by further investigating the most altered genes using the Gene Ontology and KEGG databases. Secreted cytokines, which are important in the immunological response, were also examined at the protein level, and a functional assay was performed to assess wound healing. Activated AD-MSC increased the secretion of IL-6, IL-8 and CXCL-10, and also the closure of wounds. AD-MSCs presented accelerated wound healing under inflammation conditions, suggesting that we could use this cell in clinical application. [ABSTRACT FROM AUTHOR]

ERCC1 (excision repair cross complementing-group 1) is a mammalian endonuclease that incises the damaged strand of DNA during nucleotide excision repair and interstrand cross-link repair. Ercc1-/Δ mice, carrying one null and one hypomorphic Ercc1 allele, have been widely used to study aging due to accelerated aging phenotypes in numerous organs and their shortened lifespan. Ercc1-/Δ mice display combined features of human progeroid and cancer-prone syndromes. Although several studies report cellular senescence and apoptosis associated with the premature aging of Ercc1-/Δ mice, the link between these two processes and their physiological relevance in the phenotypes of Ercc1-/Δ mice are incompletely understood. Here, we show that ERCC1 depletion, both in cultured human fibroblasts and the skin of Ercc1-/Δ mice, initially induces cellular senescence and, importantly, increased expression of several SASP (senescence-associated secretory phenotype) factors. Cellular senescence induced by ERCC1 deficiency was dependent on activity of the p53 tumor-suppressor protein. In turn, TNFα secreted by senescent cells induced apoptosis, not only in neighboring ERCC1-deficient nonsenescent cells, but also cell autonomously in the senescent cells themselves. In addition, expression of the stem cell markers p63 and Lgr6 was significantly decreased in Ercc1-/Δ mouse skin, where the apoptotic cells are localized, compared to age-matched wild-type skin, possibly due to the apoptosis of stem cells. These data suggest that ERCC1-depleted cells become susceptible to apoptosis via TNFα secreted from neighboring senescent cells. We speculate that parts of the premature aging phenotypes and shortened health- or lifespan may be due to stem cell depletion through apoptosis promoted by senescent cells.

Background The incidence of gestational diabetes mellitus (GDM) remains high, and it is prevalent all over the world. The incidence of GDM in China is up to 14.8%. Intrauterine hyperglycemia during pregnancy is a severe threat to maternal and neonatal health. Objective The aim of this study was to evaluate the effect of mindfulness-based complementary and alternative therapy for GDM. Methods This study selected 64 pregnant women with GDM undergoing routine outpatient prenatal examination in Changsha Central Hospital by using convenience sampling from March to December 2021. These patients were randomly assigned into the intervention group (n=31) and the control group (n=33) . The control group received routine psychologicalgcare, nutritional guidance and exercise guidance. The intervention group received 8 weeks of mindfulness-based complementary and alternative therapy on the basis of routine psychological care. This study collected the baseline characteristics and compared the blood sugar and inflammatory biomarkers and adipokines, followed-up neonatal outcomes (birth weight, random blood sugar) . Results The fasting blood sugar level, 1-hour blood sugar, 2-hour blood sugar and the expression of interleukin-6, interleukin-8, tumor necrosis factor-α and vaspin were lower than those of the control group (P

Rheumatoid arthritis (RA) is an autoimmune disease characterized by severe synovial inflammation and cartilage damage. Despite great progress in RA therapy, there still lacks the drugs to completely cure RA patients. Herein, we propose a reprogrammed neutrophil cytopharmaceuticals loading with TNFα-targeting-siRNA (siTNFα) as an alternative anti-inflammatory approach for RA treatment. The loaded siTNFα act as not only the gene therapeutics to inhibit TNFα production by macrophages in inflamed synovium, but also the editors to reprogram neutrophils to anti-inflammatory phenotypes. Leveraging the active tendency of neutrophils to inflammation, the reprogrammed siTNFα/neutrophil cytopharmaceuticals (siTNFα/TP/NEs) can rapidly migrate to the inflamed synovium, transfer the loaded siTNFα to macrophages followed by the significant reduction of TNFα expression, and circumvent the pro-inflammatory activity of neutrophils, thus leading to the alleviated synovial inflammation and improved cartilage protection. Our work provides a promising cytopharmaceutical for RA treatment, and puts forward a living neutrophil-based gene delivery platform.

There are controversial results regarding the value of serum IL-8 and TNFα in patients with non-specific low back pain. This study aimed to compare pro-inflammatory cytokines between patients with non-specific back pain and pain-free controls. We conducted a case–control study including 106 participants: 46 patients with chronic non-specific low back pain (G1) and 60 pain-free controls (G0). The interleukin (IL-)6, IL-8, IL-17, IL-23, IL-22, and Tumor necrosis factor α (TNFα) were measured. We collected demographic and clinical data, including age, gender, low back pain duration and radicular pain. The pain degree was assessed using the Visual Analogic Scale. The mean age was 43.17 ± 8.7 years in G1. Radicular pain was found in 37 cases with a Visual Analogic Scale of 3.03 ± 2.5 mm. The magnetic resonance imaging was performed in (G1), showing disk herniation and degenerative disk disease in 54.3% (n = 25) and 45.7% of cases (n = 21), respectively. The IL-8 was higher in G1 (18.84 ± 44.64 versus 4.34 ± 1.23 pg/mL, p :0.033). IL-8 levels correlated with TNFα (0.942, p < 10–3), IL-6 (0.490, p = 0.011) and Visual Analogic Scale Radicular-pain (r :0.297, p :0.047). IL-17 was higher in patients with restricted lumbar spine mobility (9.64 ± 20.77 versus 1.19 ± 2.54 pg/mL, p :0.014). Our results provide evidence that IL-8 and TNFα play a role in low back pain and radicular pain due to disk degeneration or herniation. These findings could potentially be used by future studies to develop new non-specific low back pain therapeutic strategies. [ABSTRACT FROM AUTHOR]

Simple Summary: Hormone receptor-positive (HR+)/HER2− breast cancer is driven by extracellular cues within the tumor microenvironment (TME) including hormonal, inflammatory and growth-stimulating signals. Our past findings indicate that a "TME Stimulation" jointly addressing these three arms induces pro-metastatic traits in HR+/HER2− breast cancer cells, primarily with the enrichment of cancer stem cells (CSCs), driving metastasis in vivo. Here, we reveal intricate roles for STAT3 as a negative and positive regulator of TME Stimulation-induced pro-metastatic effects in HR+/HER2− cells. Of note, the two transcription factors STAT3 and p65 acted in cooperativity to limit CSC enrichment, and their down-regulation has led to enriched levels of chemotherapy-resistant CSCs. Moreover, STAT3 and p65 activation were inversely connected to a CSC signature and positively associated with improved survival in patient datasets. These findings suggest that we need to carefully consider the roles of STAT3 and p65 roles in regulating TME activities in malignant diseases, in efforts to identify novel targets for therapeutic intervention. Hormone receptor-positive and HER2-negative (HR+/HER2−; luminal A) tumors are prevalent in breast cancer. Our past studies demonstrated that "TME Stimulation" (estrogen + TNFα + EGF, representing three arms of the tumor microenvironment, TME) has enriched metastasis-forming cancer stem cells (CSCs) in HR+/HER2− human breast cancer cells. Here, following information obtained by RNAseq analyses of TME-stimulated CSCs and Non-CSCs, we found that TME Stimulation has induced the activation of S727-STAT3, Y705-STAT3, STAT1 and p65. Upon TME Stimulation, stattic (STAT3 inhibitor) usage demonstrated that Y705-STAT3 activation negatively controlled CSC enrichment and epithelial-to-mesenchymal transition (EMT) traits, while inducing CXCL8 (IL-8) and PD-L1 expression. However, STAT3 knock-down (siSTAT3) had no effect on these functions; in terms of CSC enrichment, p65 had down-regulatory roles that compensated for the loss of an entire STAT3 protein. Y705-STAT3 and p65 acted additively in reducing CSC enrichment, and Y705A-STAT3 variant + sip65 has enriched chemo-resistant CSCs. Clinical data analyses revealed an inverse correlation between Y705-STAT3 + p65 phosphorylation and CSC signature in luminal A patients, and connection to improved disease course. Overall, we find regulatory roles for Y705-STAT3 and p65 in TME-stimulated HR+/HER2− tumors, with the ability to limit CSC enrichment. These findings raise concerns about using inhibitors of STAT3 and p65 as therapeutic strategies in the clinic. [ABSTRACT FROM AUTHOR]

Heat stress negatively affects the metabolism and physiology of the bovine gut. However, it is not known whether heat stress induces an inflammatory response in mesenteric lymph nodes (MLN), the primary origin of gut immune cells, and thus contributes to inflammatory processes in the circulation. Therefore, our objective was to elucidate the effects of chronic heat stress on the systemic activation of acute-phase response in blood, proinflammatory cytokine production in peripheral blood mononuclear cells (PBMC), and the activation of the toll-like receptor signaling (TLR) 2/4 pathway in MLN leucocytes and their chemokines and chemokine receptor profiles in Holstein cows. Primiparous Holstein cows (n = 30; 169 ± 9 d in milk) were exposed to a temperature-humidity index (THI) of 60 [16°C, 63% relative humidity (RH)] for 6 d. Thereafter, cows were evenly assigned to 3 groups: heat-stressed (HS; 28°C, 50% RH, THI = 76), control (CON; 16°C, 69% RH, THI = 60), or pair-feeding (PF; 16°C, 69% RH, THI = 60) for 7 d. On d 6, PBMC were isolated and on d 7 MLN. Plasma haptoglobin, TNFα, and IFNγ concentrations increased more in HS than CON cows. Concomitantly, TNFA mRNA abundance was higher in PBMC and MLN leucocytes of HS than PF cows, whereas IFNG mRNA abundance tended to be higher in MLN leucocytes of HS than PF cows, but not for chemokines (CCL20, CCL25) or chemokine receptors (ITGB7, CCR6, CCR7, CCR9). Furthermore, the TLR2 protein expression tended to be more abundant in MLN leucocytes of HS than PF cows. These results suggest that heat stress induced an adaptive immune response in blood, PBMC, and MLN leukocytes involving the acute-phase protein haptoglobin, proinflammatory cytokine production, and TLR2 signaling in MLN leucocytes. However, chemokines regulating the leucocyte trafficking between MLN and gut seem not to be involved in the adaptive immune response to heat stress. [ABSTRACT FROM AUTHOR]

We prepared three-dimensional (3-D) organoids of human stomach cancers and examined the correlation between the tumorigenicity and cytotoxicity of Helicobacter pylori (H. pylori). In addition, the effects of hepatoma-derived growth factor (HDGF) and tumor necrosis factor (TNFα) on the growth and invasion activity of H. pylori-infected gastric cancer organoids were examined. Cytotoxin-associated gene A (CagA)-green fluorescence protein (GFP)-labeled H. pylori was used to trace the infection in gastric organoids. The cytotoxicity of Cag encoded toxins from different species of H. pylori did not affect the proliferation of each H. pylori-infected cancer organoid. To clarify the role of HDGF and TNFα secreted from H. pylori-infected cancer organoids, we prepared recombinant HDGF and TNFα and measured the cytotoxicity and invasion of gastric cancer organoids. HDGF controlled the growth of each organoid in a species-specific manner of H. pylori, but TNFα decreased the cell viability in H. pylori-infected cancer organoids. Furthermore, HDGF controlled the invasion activity of H. pylori-infected cancer organoid in a species-dependent manner. However, TNFα decreased the invasion activities of most organoids. We found different signaling of cytotoxicity and invasion of human gastric organoids in response to HDGF and TNFα during infection by H. pylori. Recombinant HDGF and TNFα inhibited the development and invasion of H. pylori-infected gastric cancer differently. Thus, we propose that HDGF and TNFα are independent signals for development of H. pylori-infected gastric cancer. The signaling of growth factors in 3-D organoid culture systems is different from those in two-dimensional cancer cells. [ABSTRACT FROM AUTHOR]

The aim of the present study was to determine the effects of carboxymethyl chitosan (CMC) on titanium particles-induced oxidative stress in mouse RAW264.7 macrophages. The mouse RAW264.7 macrophages were divided into four groups: (i) the control group; (ii) the CMC group received stimulation of CMC for 4 h; (iii) the titanium particles group received stimulation of titanium particles for 12 h; and (iv) the CMC group received pre-stimulation of CMC hydrogels for 4 h followed by treatment of titanium particles for 12 h. Afterwards, reactive oxygen species (ROS) level in the cells was measured by flow cytometry. A spectrophotometer was used to measure the activities of oxidases and antioxidant enzymes. Fluorescence quantitative PCR was performed to analyze mRNA levels of enzymes and tumor necrosis factor α (TNF-α). ELISA was used to detect the mass concentration of TNF-α after indicated treatment. CMC effectively suppressed titanium particles-induced oxidative stress in RAW264.7 cells, as evidenced by the decrease in intracellular ROS level, the transcription of oxidases, and TNF-α concentration as well as the increase in the transcription of antioxidant enzymes. CMC exerts a protective impact against wear particles-induced oxidative stress and reduces the release of TNF-α in RAW264.7 cells. [ABSTRACT FROM AUTHOR]

Abstract Introduction Multiple sclerosis (MS) is the most common demyelinating disease of the central nervous system. However, the limitations of available therapeutic strategies are frustrating, both in terms of their low efficacy and multiple side effects. Previous studies showed that natural compounds such as Chalcones possess neuroprotective effects on neurodegenerative disorders. However, few studies have so far been published on the potential effects of Chalcones on treating demyelinating disease. The present study was designed to investigate the effects of Chalcones from Ashitaba (ChA) on cuprizone‐induced noxious changes in the C57BL6 mice model of MS. Methods The mice received normal diets (Control group: CNT), or Cuprizone‐supplemented diets either without ChA (Cuprizone group: CPZ) or with low or high (300, 600 mg/kg/day) doses of ChA (ChA‐treated groups: CPZ+ChA300/600). Brain‐derived neurotrophic factor (BDNF) and tumor necrosis factor alpha (TNFα) levels, demyelination scores in the corpus callosum (CC), and cognitive impairment were evaluated using the enzyme‐linked immunosorbent assay, histological, and Y‐maze tests, respectively. Results The findings showed that ChA Co‐treatment significantly reduced the extent of demyelination in the CC and the serum and brain levels of TNFα in the ChA‐treated groups compared to the CPZ group. Besides, treatment with a higher dose of ChA significantly improved the behavioral responses and BDNF levels in the serum and brain of the CPZ+ChA600 group when compared with the CPZ group. Conclusion The present study provided evidence for the neuroprotective effects of ChA on cuprizone‐induced demyelination and behavioral dysfunction in C57BL/6 mice, possibly by modulating TNFα secretion and BDNF expression.

The current gold standard to treat large cartilage defects is autologous chondrocyte transplantation (ACT). As a new surgical method of cartilage regeneration, minced cartilage implantation (MCI) is increasingly coming into focus. The aim of this study is to investigate the influence of chondrogenesis between isolated and cultured chondrocytes compared to cartilage chips in a standardized inflammation model with the proinflammatory cytokine TNFα. Articular chondrocytes from bovine cartilage were cultured according to the ACT method to passage 3 and transferred to spheroid culture. At the same time, cartilage was fragmented (3) to produce cartilage chips. TNFα (20 ng/mL) was supplemented to simulate an inflammatory process. TNFα had a stronger influence on the passaged chondrocytes compared to the non-passaged ones, affecting gene expression profiles differently between isolated chondrocytes and cartilage chips. MCI showed less susceptibility to TNFα, with reduced IL-6 release and less impact on inflammation markers. Biochemical and histological analyses supported these findings, showing a greater negative influence of TNFα on the passaged pellet cultures compared to the unpassaged cells and MCI constructs. This study demonstrated the negative influence of TNFα on chondrogenesis in a chondrocyte spheroid culture and cartilage fragment model. Passaged chondrocytes are more sensitive to cytokine influences compared to non-passaged cells and chondrons. This suggests that MCI may have superior regeneration potential in osteoarthritic conditions compared to ACT. Further investigations are necessary for the translation of these findings into clinical practice.

Background: Our previous studies demonstrated that sour cherry anthocyanins (AC) reduce the salivary count of Streptococcus mutans and inhibit salivary amylase activity within 30 minutes after chewing AC gum. AC gum and changing toothbrushes after scaling reduced the Gram-negative species in the unstimulated salivary microbiota. The present study examined the effect of AC gums on salivary factors, including changes in microbiome. Methods: The study was conducted over three weeks with two groups; young adults (18–30) and adults (30–45). Ten participants changed their toothbrushes, while the other 10 participants did not change after the control period. After scaling, all participants received three doses of AC gum daily. The salivary mRNA and protein levels of cytokines, mucins, melatonin, and the microbiota of unstimulated and stimulated saliva were determined by polymerase chain reaction, enzyme-linked immunosorbent assay, and 16S rRNA gene sequencing. Results: Significantly higher levels of tumor necrosis factor α (TNFα), interleukin-1β (IL-1β), mucin5B (MUC5B), mucin7 (MUC7), and melatonin were detected in stimulated saliva. Correlation analysis of these factors with the microbiota showed positive correlations with the genera Lachnospiraceae, Eikenella, Saccharibacteria_(TM7), Streptococcus, Prevotella, and Haemophilus. Conclusions: AC chewing gum has a beneficial effect on the composition of the oral microbiome, and toothbrush replacement leads to changes in the levels of salivary pro-inflammatory cytokines.

Abstract Background Rheumatoid arthritis (RA) patients present with abnormal methylation patterns in their fibroblast-like synoviocytes (FLS). Given that DNA demethylation is critical for producing DNA methylation patterns, we hypothesized that DNA demethylation may facilitate RA progression. Therefore, we designed this study to examine the role of DNA dioxygenase family, Ten-Eleven translocation (TET1/2/3), in the pathological process of RA. Methods Synovial tissues and FLS were obtained from patients with RA and Osteoarthritis. K/BxN serum-induced arthritis was induced in Wild-type (WT) and TET3 heterozygous-deficient (TET3 +/− ) C57BL/6 mice. Results We found that both TET3 and 5-hydroxymethylcytosine (5hmC) were upregulated in synovitis tissues from RA patients and confirmed this upregulation in the cultured FLS derived from synovitis tissues. Tumor necrosis factor α (TNFα) upregulated TET3 and 5hmC levels in cultured FLS, and the stimulated FLS exhibited high cell mobility with increased transcription of cellular migration-related factors such as C-X-C motif chemokine ligand 8 (CXCL8) and C-C motif chemokine ligand 2 (CCL2) in a TET3-dependent manner. In addition, TET3 haploinsufficiency lowered RA progression in a mouse model of serum-induced arthritis. Conclusions Based on these findings, we can assume that TET3-mediated DNA demethylation acts as an epigenetic regulator of RA progression.

Angiogenesis, a process characterized by the formation of new blood vessels from pre‐existing ones, is a crucial step in tumor growth and dissemination. Given the ability of tumors to interfere with multiple or different molecular pathways to promote angiogenesis, there is an increasing need to therapeutically block tumor progression by targeting multiple antiangiogenic pathways. Natural polyphenols present health‐protective properties, which are likely attributed to their ability to activate multiple pathways involved in inflammation, carcinogenesis, and angiogenesis. Recently, increased attention has been addressed to the ability of flavonoids, the most abundant polyphenols in the diet, to prevent cancer by suppressing angiogenesis. Here we investigate the mechanisms by which xanthohumol (the major prenylated flavonoid of the hop plant Humulus lupulus L.) and nobiletin (flavonoid from red‐orange Citrus sinensis) can modulate the effects of Tumor Necrosis Factor‐α (TNF‐α) on human umbilical vein endothelial cells (HUVEC). The results reported in this paper show that xanthohumol and nobiletin pretreatment of HUVEC inhibits the effects induced by TNF‐α on cell migration, invasion capability, and colon cancer cell adhesion on the endothelial monolayer. Moreover, the pretreatment reduces metalloproteinases and adhesion molecules' expression. Finally, our results highlight that xanthohumol and nobiletin can counteract the effects of TNF‐α on angiogenesis and invasiveness, mainly through Vascular Endothelial Growth Factor and NF‐κB pathways. Since angiogenesis plays an important pathological role in the progression of several diseases, our findings may provide clues for developing xanthohumol and nobiletin as therapeutic agents against angiogenesis‐associated diseases. [ABSTRACT FROM AUTHOR]

Objective To investigate the efficacy of modified Xiaochaihu decoction combined with oseltamivir phosphate granules in the treatment of influenza in children. Methods A total of 130 influenza children admitted to this hospital from May 2020 to May 2021 were selected as the study objects, and were divided into the control group and the treatment group according to random number table method, with 65 cases in each group. The control group was treated with oseltamivir phosphate granules, and the treatment group was treated with modified Xiaochaihu decoction on the basis of the control group. The two groups were treated continuously for one week. The disappearance time of symptoms and signs, blood routine indexes, clinical efficacy, serum levels of interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α), and adverse reaction were compared between the two groups. Results The disappearance time of fever, cough, sore throat and fatigue in the treatment group was significantly shorter than that in the control group, and the differences was statistically significant(P<0. 01). After treatment, the white blood cell count and lymphocyte count of the two groups were significantly reduced, and the treatment group was significantly less than the control group, the differences were statistically significant(P<0. 01). The total effective rate of the treatment group was significantly higher than that of the control group, the difference was statistically significant(P<0. 05). After treatment, the serum levels of IL-6 and TNF-α in the two groups were significantly decreased, and the treatment group was significantly lower than the control group, the differences were statistically significant(P<0. 01). The incidence of adverse reaction in the treatment group was significantly lower than that in the control group, and the difference was statistically significant(P<0. 05). Conclusion The efficacy of modified Xiaochaihu decoction combined with oseltamivir phosphate granules in treating influenza children is significant. It can promote the improvement of the symptoms and signs in children with influenza, and inhibit inflammation. [ABSTRACT FROM AUTHOR]

Rheumatoid arthritis (RA) is an autoimmune disease characterized by severe synovial inflammation and cartilage damage. Despite great progress in RA therapy, there still lacks the drugs to completely cure RA patients. Herein, we propose a reprogrammed neutrophil cytopharmaceuticals loading with TNF α -targeting-siRNA (siTNF α) as an alternative anti-inflammatory approach for RA treatment. The loaded siTNF α act as not only the gene therapeutics to inhibit TNF α production by macrophages in inflamed synovium, but also the editors to reprogram neutrophils to anti-inflammatory phenotypes. Leveraging the active tendency of neutrophils to inflammation, the reprogrammed siTNF α /neutrophil cytopharmaceuticals (siTNF α /TP/NEs) can rapidly migrate to the inflamed synovium, transfer the loaded siTNF α to macrophages followed by the significant reduction of TNF α expression, and circumvent the pro-inflammatory activity of neutrophils, thus leading to the alleviated synovial inflammation and improved cartilage protection. Our work provides a promising cytopharmaceutical for RA treatment, and puts forward a living neutrophil-based gene delivery platform. Reprogrammed siRNA/neutrophil cytopharmaceuticals actively targeted to inflamed joints, effectively inhibited the TNF α production, thus leading to the alleviated synovial inflammation and improved cartilage protection, providing promising cytopharmaceuticals for rheumatoid arthritis (RA) treatment. [Display omitted] [ABSTRACT FROM AUTHOR]

Inflammatory bowel disease, Metabolic bone disease, Hypophosphatemia, Oncogenic osteomalacia, Tumor necrosis factor , Fibroblast growth factor 23 Although FGF-23 levels decreased during remission, they remained higher than control patients without IBD [[6]], suggesting that IBD itself may portend a worsened prognosis in patients who simultaneously develop FGF-23 secreting tumors. 2 Histology showing proliferation of spindled to ovoid and epithelioid cells in sheets, suggestive of phosphaturic mesenchymal tumor Discussion Tumor-induced osteomalacia (TIO), also known as oncogenic osteomalacia, is a rare paraneoplastic syndrome characterized by chronic normocalcemic hypophosphatemia and impaired vitamin D metabolism [[1]]. Keywords: Inflammatory bowel disease; Metabolic bone disease; Hypophosphatemia; Fibroblast growth factor 23; Oncogenic osteomalacia; Tumor necrosis factor EN Inflammatory bowel disease Metabolic bone disease Hypophosphatemia Fibroblast growth factor 23 Oncogenic osteomalacia Tumor necrosis factor 54 57 4 01/31/23 20230101 NES 230101 Case Presentation A 34-year-old male was hospitalized with severe abdominal pain, non-bloody diarrhea and an intra-abdominal abscess adjacent to the terminal ileum that did not resolve with antibiotic therapy. [Extracted from the article]

Neuronal apoptosis is considered to be a critical cause of Alzheimer's disease (AD). Recently, meloxicam has shown neuroprotective effects; however, the inherent mechanisms are highly overlooked. Using APP/PS1 transgenic (Tg) mice as in vivo animal models, we found that meloxicam inhibits apoptosis in neurons by deactivating tumor necrosis factor receptor superfamily member 25 (TNFRSF25), leading to the suppression of the expression of fas-associated protein with death domain (FADD) and the cleavage of DNA fragmentation factor subunit α (DFFA) and cysteine aspartic acid protease-3 (caspase 3) via β-amyloid protein (Aβ)-depressing mechanisms. Moreover, the meloxicam treatment blocked the effects of β-amyloid protein oligomers (Aβo) on stimulating the synthesis of tumor necrosis factor α (TNF-α) and TNF-like ligand 1A (TL1A) in neuroblastoma (N) 2a cells. TNF-α and TL1A induce apoptosis in neurons via TNFR- and TNFRSF25-dependent caspase 3-activating mechanisms, respectively. Knocking down the expression of TNFRSF25 blocked the effects of TL1A on inducing apoptosis in neurons by deactivating the signaling cascades of FADD, caspase 3, and DFFA. Consistently, TNFRSF25 shRNA blocked the effects of Aβo on inducing neuronal apoptosis, which was corroborated by the efficacy of meloxicam in inhibiting Aβo-induced neuronal apoptosis. By ameliorating neuronal apoptosis, meloxicam improved memory loss in APP/PS1 Tg mice. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to assess the anti-rheumatic potential of Dodonaea viscosa and to evaluate its bioactive small molecules for their beneficial effects in the management of rheumatoid arthritis.Methods:In vitro bioactivity assays were performed to assess the healing potential of D. viscosa and statistical analysis was performed by using the linear regression technique. In silico analysis was performed to identify the key inhibitors of the disease to target TNF-α. The plant extract was prepared using ethanol solvent via the Soxhlet method. Phytochemical and bioactivity testing was performed. Gas chromatography–mass spectrometry (GC-MS) analysis was conducted for bioactive plant compounds. Disease-specific target was shortlisted by HUB gene analysis. Molecular docking and molecular dynamic simulations were run for validation of the results.Results: Phytochemical studies verified the presence of phenols, flavonoids, steroids, sterols, saponins, coumarins, tannins, and terpenoids. The significant antioxidant potential of plant extract was evaluated by the DPPH and Ferric Reducing Antioxidant Power (FRAP) assays, while the anti-inflammatory potential was evaluated by the protein denaturation and Human Red Blood Cell (HRBC) membrane stabilization assays. In silico studies revealed that nine of the 480 compounds found in D. viscosa (ethanol extract) had drug-like properties. Tumor necrosis factor alpha (TNF-α) was selected as a key disease gene through HUB gene analysis. Results of molecular docking and MD simulation analysis demonstrated that 4-(1-hydroxy-3-oxo-1H-isoindol-2-yl) benzoic acid (PubChemID 18873897), had the best binding affinity with TNF-α amongst all nine compounds.Conclusion: 4-(1-hydroxy-3-oxo-1H-isoindol-2-yl) benzoic acid (PubChemID 18873897), have the potential to be a good small molecule inhibitor of TNF-α against rheumatoid arthritis.

Excessive release of inflammatory cytokines has been considered as a major cause of chronic inflammation, resulting in intestinal barrier disruption that leads to inflammatory bowel disease (IBD). Tumor necrosis factor α (TNFα) is one of the well-known inflammatory cytokines that activates formation of NLRP3 inflammasome, thus resulting in excessive secretion of inflammatory cytokines causing IBD. Although immunoproteasome inhibitors have been reported to inhibit inflammatory cytokine release, immunoproteasome inhibition has not yet been addressed for attenuation of NLRP3 inflammasome activity in intestinal epithelial cell. Here, we observed that NLRP3 inflammasome assembly was attenuated by peptide epoxyketone YU102, a LMP2 subunit immunoproteasome inhibitor, in intestinal epithelial cell. YU102 also inhibited maturation of active caspase-1 and secretion of IL-1β, which are subsequent inflammatory cascade after the formation of NLRP3 inflammasome. Progression of epithelial-mesenchymal transition and increase of cellular permeability, which were induced by TNFα, were also suppressed through inhibition of immunoproteasome. Furthermore, we found that YU102 does not inhibit degradation of IкBα and its following NF-кB activation that leads to transcription of NLRP3. These findings suggest that inhibition of immunoproteasome with YU102 offers a potential therapeutic premise for prevention of TNFα-induced chronic inflammation through attenuation of NLRP3 inflammasome assembly. • TNFα induces NLRP3 inflammasome and immunoproteasome in intestinal epithelial cells. • Immunoproteasome inhibition with YU102 attenuates assembly of NLRP3 inflammasome. • Immunoproteasome inhibition suppresses progression of EMT and cellular permeability. • Immunoproteasome does not entail NF-κB pathway for NLRP3 inflammasome assembly. [ABSTRACT FROM AUTHOR]

Rosai-Dorfman disease (RDD) is a rare histiocytic proliferative disorder characterized by lymphadenopathy and extra-nodal manifestations. Some patients with RDD require systemic treatment, but there is no consensus on the treatment strategy owing to its extreme rarity. Overexpression of tumor necrosis factor α (TNF-α) has been reported in lesions of patients with RDD and is thought to be involved in its pathogenesis. We herein report the first case of RDD with cutaneous involvement and arthritis that was successfully treated with methotrexate and infliximab. This case highlights the potential efficacy of anti-TNF-α therapy for RDD, offering a novel treatment option for this rare condition.

Adipose-derived mesenchymal stem cells are increasingly being used in regenerative medicine as cell therapy targets, including in the treatment of burns and ulcers. The regenerative potential of AD-MSCs and some of their immunological properties are known from in vitro studies; however, in clinical applications, cells are used in non-ideal conditions and can behave differently in inflammatory environments, affecting the efficacy and outcome of therapy. Our aim was to investigate and map the pathways that the inflammatory microenvironment can induce in these cells. High-throughput gene expression assays were performed on AD-MSCs activated with LPS and TNFα. Analysis of RNA-Seq data showed that control, LPS-treated and TNFα-treated samples exhibited distinct gene expression patterns. LPS treatment increased the expression of 926 genes and decreased the expression of 770 genes involved in cell division, DNA repair, the cell cycle, and several metabolic processes. TNFα treatment increased the expression of 174 genes and decreased the expression of 383 genes, which are related to cell division, the immune response, cell proliferation, and differentiation. We also map the biological pathways by further investigating the most altered genes using the Gene Ontology and KEGG databases. Secreted cytokines, which are important in the immunological response, were also examined at the protein level, and a functional assay was performed to assess wound healing. Activated AD-MSC increased the secretion of IL-6, IL-8 and CXCL-10, and also the closure of wounds. AD-MSCs presented accelerated wound healing under inflammation conditions, suggesting that we could use this cell in clinical application.

Background: The hypothalamic-pituitary-adrenal (HPA) axis, inflammatory processes and neurotrophic factor systems are involved in pathogenesis of both epilepsy and depressive disorders. The study aimed to explore these systems in patients with focal epilepsy (PWE, n = 76), epilepsy and comorbid depression (PWCED n = 48), and major depressive disorder (PWMDD, n = 62) compared with healthy controls (HC, n = 78). Methods: Parameters of the HPA axis, neurotrophic factors, and TNF-α were measured in blood serum along with the hemogram. Results: Serum cortisol level was augmented in PWE, PWCED, and PWMDD compared with HC and was higher in PWMDD than in PWE. Serum cortisol negatively correlated with Mini–Mental State Examination (MMSE) score in PWE, and positively with depression inventory–II (BDI-II) score in PWMDD. Only PWMDD demonstrated elevated plasma ACTH. Serum TNF-α, lymphocytes, and eosinophils were augmented in PWMDD; monocytes elevated in PWE and PWCED, while neutrophils were reduced in PWE and PWMDD. Serum BDNF was decreased in PWE and PWCED, CNTF was elevated in all groups of patients. In PWE, none of above indices depended on epilepsy etiology. Conclusions: The results confirm the involvement of HPA axis and inflammatory processes in pathogenesis of epilepsy and depression and provide new insights in mechanisms of epilepsy and depression comorbidity. [ABSTRACT FROM AUTHOR]

Both hypernatremia and an abnormal immune response may increase hospital mortality in patients with sepsis. This study examined the association of hypernatremia with abnormal immune response and mortality in 520 adult patients with sepsis in an intensive care unit (ICU). We compared the mortality and ex vivo lipopolysaccharide (LPS)-induced inflammatory response differences among patients with hyponatremia, eunatremia, and hypernatremia, as well as between patients with acquired hypernatremia on ICU day 3 and those with sustained eunatremia over first three ICU days. Compared with eunatremia or hyponatremia, hypernatremia led to higher 7 day, 14 day, 28 day, and hospital mortality rates (p = 0.030, 0.009, 0.010, and 0.033, respectively). Compared with sustained eunatremia, acquired hypernatremia led to higher 7, 14, and 28 day mortality rates (p = 0.019, 0.042, and 0.028, respectively). The acquired hypernatremia group nonsignificantly trended toward increased hospital mortality (p = 0.056). Day 1 granulocyte colony-stimulating factor (G-CSF) and tumor necrosis factor (TNF) α levels were relatively low in patients with hypernatremia (p = 0.020 and 0.010, respectively) but relatively high in patients with acquired hypernatremia (p = 0.049 and 0.009, respectively). Thus, in ICU-admitted septic patients, hypernatremia on admission and in ICU-acquired hypernatremia were both associated with higher mortality. The higher mortality in patients with hypernatremia on admission was possibly related to the downregulation of G-CSF and TNF-α secretion after endotoxin stimulation. Compared to sustained eunatremia, acquired hypernatremia showed immunoparalysis at first and then hyperinflammation on day 3. [ABSTRACT FROM AUTHOR]

Early pregnancy loss (EPL) is a relatively common pathology of which almost 50% of cases remain idiopathic. In the search for novel biomarkers, differential scanning calorimetry (DSC) is intensively used to characterize the thermodynamic behavior of blood plasma/serum proteome in health and disease. Herein, for the first time, we investigate the DSC denaturation profiles of blood plasma derived from patients suffering EPL compared to healthy pregnant and non-pregnant women. Data analysis reveals that 58% of the EPL thermograms differ significantly from those of healthy pregnant women. Thermal stabilization of a fraction of albumin-assigned transition with concomitant suppression of the major and enhancement of the globulin-assigned transition are characteristic features of EPL calorimetric profiles that could be used as a new indicator of a risk pregnancy. The presented results suggest an altered composition or intermolecular interactions of the plasma proteome of women with EPL. In addition, the alterations of the EPL thermograms correlate with the increased blood levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) and a higher prevalence of the polymorphism in the plasminogen activator inhibitor type-1 (PAI-1) gene, suggesting an expression of an overall enhanced immune response. The concomitant changes in plasma thermograms confirm the potential of the DSC approach for distinguishing changes in the pathological state of the blood plasma proteome. [ABSTRACT FROM AUTHOR]

Tumor necrosis factor α (TNF‐α) inhibitors have shown great success in the treatment of autoimmune diseases. However, to date, approved drugs targeting TNF‐α are restricted to biological macromolecules, largely due to the difficulties in using small molecules for pharmaceutical intervention of protein–protein interactions. Herein the power of a natural product‐enriched DNA‐encoded library (nDEL) is exploited to identify small molecules that interfere with the protein–protein interaction between TNF‐α and the cognate receptor. Initially, to select molecules capable of binding to TNF‐α , "late‐stage" DNA modification method is applied to construct an nDEL library consisted of 400 sterically diverse natural products and pharmaceutically active chemicals. Several natural products, including kaempferol, identified not only show direct interaction with TNF‐α, but also lead to the blockage of TNF‐α/TNFR1 interaction. Significantly, kaempferol attenuates the TNF‐α signaling in cells and reduces the 12‐O‐tetradecanoylphorbol‐13‐acetateinduced ear inflammation in mice. Structure‐activity‐relationship analyses demonstrate the importance of substitution groups at C‐3, C‐7, and C‐4' of kaempferol. The nDEL hit, kaempferol, represents a novel chemical scaffold capable of specifically recognizing TNF‐α and blocking its signal transduction, a promising starting point for the development of a small molecule TNF‐α inhibitor for use in the clinical setting. [ABSTRACT FROM AUTHOR]