TNF-α Concentrations Research Articles

Indoleamine 2, 3-dioxygenase Regulates the Differentiation of T Lymphocytes to Promote the Growth of Gastric Cancer Cells through the PI3K/Akt/mTOR Pathway.

To investigate the regulatory mechanism of indoleamine 2, 3-dioxygenase (IDO) in T lymphocyte differentiation and its role in promoting the growth of gastric cancer (GC) cells through the PI3K/Akt/mTOR pathway. GC cell lines (MFC and NCI-N87) and PBMC cells were co-cultured and IDO inhibitor 1-methyl-tryptophan (1-MT) was added. The proliferation was detected by CCK-8, the apoptosis was detected by flow cytometry, and the contents of TNF-α, IL-1β, IL-6, IL-8, and INF-γ were detected by ELISA. The expression levels of PI3K, p-PI3K, Akt, p-Akt, mTOR, and p-mTOR were tested using Western blot, and the proportion of CD4+/CD8+, CD4+CD25+Foxp3+Treg cells was detected by flow cytometry. C57BL/6 mice were used to establish the MFC GC mouse model and treated with 1-MT. The changes in body weight and tumor diameter were measured. Ki-67, CD4+, CD8+, and CD25+ expressions were detected by immunohistochemistry. IDO promoted the proliferation of MFC and NCI-N87 cells, inhibited apoptosis, and decreased the levels of TNF-α, IL-1β, IL-6, IL-8, and INF-γ in the supernatant after co-culture with BPMC. The expressions of p-AKT, p-mTOR, and p-PI3K increased after 1-MT treatment. The proportion of CD4+/CD8+ cells was increased and the proportion of Treg cells was decreased in PBMC cells after the addition of 1-MT. Overexpression of IDO suppressed T cells differentiation by inhibiting the PI3K/Akt/mTOR pathway. In vivo, 1-MT treatment reduced the tumor size and weight, increased CD4+ and CD8+ positive area proportion, and decreased Ki-67 and CD25+ positive area proportion. Co-culture of GC cells and immune cells promotes the proliferation of GC cells and inhibits apoptosis, which can be reversed by 1-MT. IDO may suppress the proliferation of T lymphocyte through inhibiting the PI3K/Akt/mTOR signaling pathway. This provides new evidence for the potential of exploiting IDO inhibitors for GC treatment.

Extracellular vesicles from mesenchymal stem cells improve neuroinflammation and neurotransmission in hippocampus and cognitive impairment in rats with mild liver damage and minimal hepatic encephalopathy

BackgroundPatients with steatotic liver disease may show mild cognitive impairment. Rats with mild liver damage reproduce this cognitive impairment, which is mediated by neuroinflammation that alters glutamate neurotransmission in the hippocampus. Treatment with extracellular vesicles (EV) from mesenchymal stem cells (MSC) reduces neuroinflammation and improves cognitive impairment in different animal models of neurological diseases. TGFβ in these EVs seems to be involved in its beneficial effects. The aim of this work was to assess if MSCs-EVs may improve cognitive impairment in rats with mild liver damage and to analyze the underlying mechanisms, assessing the effects on hippocampal neuroinflammation and neurotransmission. We also aimed to analyze the role of TGFβ in the in vivo effects of MSCs-EVs.MethodsMale Wistar rats with CCl4-induced mild liver damage were treated with EVs from unmodified MSC or with EVs derived from TGFβ—silenced MSCs and its effects on cognitive function and on neuroinflammation and altered neurotransmission in the hippocampus were analysed.ResultsUnmodified MSC-EVs reversed microglia activation and TNFα content, restoring membrane expression of NR2 subunit of NMDA receptor and improved object location memory. In contrast, EVs derived from TGFβ—silenced MSCs did not induce these effects but reversed astrocyte activation, IL-1β content and altered GluA2 AMPA receptor subunit membrane expression leading to improvement of learning and working memory in the radial maze.ConclusionsEVs from MSCs with TGFβ silenced induce different effects on behavior, neuroinflammation and neurotransmitter receptors alterations than unmodified MSC-EVs, indicating that the modification of TGFβ in the MSC-EVs has a notable effect on the consequences of the treatment. This work shows that treatment with MSC-EVs improves learning and memory in a model of mild liver damage and MHE in rats, suggesting that MSC-EVs may be a good therapeutic option to reverse cognitive impairment in patients with steatotic liver disease.

Pericoronary adipose tissue radiomics features as imaging markers for coronary artery disease risk assessment: insights from gene expression analysis

AimsThis study aimed to explore the correlation between radiomics features of pericoronary adipose tissue (PCAT) and gene expression in patients with coronary artery disease (CAD), with the goal of identifying novel imaging biomarkers for evaluating CAD.MethodsBetween November 2021 and May 2022, data were collected from 60 patients diagnosed with CAD who underwent coronary artery bypass grafting (CABG) and coronary computed tomography angiography (CCTA). Samples of PCAT, three additional adipose tissue types, and peripheral venous blood were analysed. Radiomics features of PCAT were extracted. Gene expression in adipose tissues and serum was quantified via RT-qPCR, immunohistochemistry and ELISA. The correlations between the radiomics features and genes were analysed.ResultsGene expression analysis revealed significantly elevated levels of CD31, MCP-1, and leptin in PCAT compared with other adipose tissues, and the radiomics features of PCAT have a strong correction with the expression of CD31 and MCP-1. At the systemic level, serum analysis revealed increased concentrations of TNF-α, IL-6, CD31, COL1A1, and resistin, with notable decreases in ADP in CAD patients relative to controls. Notably, CD31, ADP, IL-6, and resistin were significantly corrlated with PCAT texture features, whereas TNF-α was correlated with first-order features.ConclusionsOur findings demonstrated a significant correlation between PCAT radiomics features and gene expression patterns in CAD patients. These features effectively reflect the pathological state of tissues and hold potential as innovative imaging biomarkers. By leveraging PCAT radiomics, clinicians may gain valuable insights for advanced evaluation and management of CAD in later stages.

Exercise preconditioning and resveratrol reduce the susceptibility of rats with obstructive jaundice to endotoxin and alleviate lung injury

IntroductionEndotoxemia is a common issue for patients with biliary obstruction. The lung is the most affected organ by endotoxins. Exercise training can alleviate lipopolysaccharide (LPS)-induced lung inflammation and resveratrol has biological effects similar to exercise. In this study, we evaluated the protective effects of exercise preconditioning, resveratrol, and their combination on LPS-induced lung injury and mortality in rats with obstructive jaundice.MethodsEndotoxemia was simulated in rats by common bile duct ligation (CBDL) and intraperitoneal injection of low-dose LPS. The treatment groups were pretreated with exercise and/or resveratrol to assess their effects on lung injury and mortality. Immunohistochemistry, immunofluorescence, and ELISA were subsequently used to evaluate the impact of exercise and/or resveratrol on inflammation in lung tissue and bronchoalveolar lavage fluid.ResultsWe found that even in the early stages, compared to sham-LPS rats, low-dose LPS induced excessive systemic inflammatory responses in CBDL rats, as evidenced by a significant increase in TNF-α and IL-6, severe lung inflammation, lung injury, and higher mortality rates, indicating that cholestasis increased rats’ susceptibility to endotoxins. Exercise training reduced neutrophil infiltration in the lungs of model rats and IL-6 levels in bronchoalveolar lavage fluid. Both exercise training and resveratrol exhibited synergistic effects in reducing macrophage accumulation in lung tissues, lowering TNF-α and IL-6 levels in the lungs, and decreasing TNF-α concentration in bronchoalveolar lavage fluid. Additionally, exercise and combined interventions both significantly increased the expression of IL-10. The interventions induced a marked improvement in lung tissue pathological damage and lung edema in model rats and prolonged the survival time of rats with obstructive jaundice.DiscussionThis study demonstrates that exercise preconditioning and/or resveratrol can significantly reduce rats’ susceptibility to endotoxins after CBDL and alleviate lung injuries through their anti-inflammatory effects, thereby decreasing the mortality risk.

Effect of milk thistle (Silybum marianum L.) oil on pro-inflammatory cytokines, acute phase proteins, rumen metagenomic profile, rumen fluid variables and performance in calves.

The aim of this study was to determine the effect of milk thistle (Silybum marianumL., SM) oil supplementation on proinflammatory cytokines, acute phase proteins, rumen metagenomic profile, rumen fluid variables and performance during the milk feeding period of Holstein calves. In the present study, 24 calves that consumed quality and sufficient amount of colostrum (≥50 mg/ml IgG) after birth were divided into three groups, with 8 animals in each group (4 males + 4 females). Individually fed calves were given added SM oil as 0 µL/day/calf (Control group, SM0), 100 µL/day/calf (SM100) and 200 µL/day/calf (SM200). The ration containing concentrated feed mix (90%) + wheat straw (10%) was offered to the calves. SM oil did not change the ammonia nitrogen and pH values of the rumen fluid of calf (P>0.05). The molarities of PA, IBA, IVA and BSCFA in the rumen fluid increased linearly with SM oil dose (P<0.05). The relative abundance of Firmicutes linearly increased and the relative abundance of Actinobacteriota decreased with the addition of SM oil (P<0.05). Relative abundances of Erysipelotrichaceae_UCG_002, Eubacterium_coprostanoligenes_group, Clostridia_UCG_014, Lachnospiraceae_Unknown_1, Lachnospiraceae_NK3A20_group, Shuttleworthia, Selenomonadaceae_Uncultured_1, Rikenellaceae_RC9_gut_group and Succinivibrionaceae_UCG_001 linearly increased with SM oil (P<0.05). Relative abundances of Methanobrevibacter, Acetitomaculum, Olsenella and Megasphaera in calf rumen fluid linearly decreased with SM oil (P<0.05). Concentrations of TNF-α, IFN-ɣ, and SAA of calf' serum at weaning stage linearly decreased with SM oil doses (P<0.05). Serum IgA concentration increased with 100 µL SM oil /day (P<0.05). As a result, the addition of SM oil to calves has the potential to reduce the immune suppression of calves during the milk feeding period and at weaning time, has a positive effect on the microbiome involved in starch and protein catabolism in the rumen fluid, and increases the fermentation end products (PA, IVA and BA). Milk thistle oil has an inhibitory effect on methanogenic archaea and can be used as an anti-methanogenic feed additive and will contribute to the effective use of feed energy.

Better cardiometabolic/inflammatory profile is associated with differences in the supraclavicular adipose tissue activity of individuals with T2DM.

Brown adipose tissue (BAT), located in the supraclavicular region, has been associated with a better cardiometabolic profile and reduced risk of developing non-communicable chronic diseases (NCD), in addition to being associated with a healthier phenotype in obesity. However, it is unknown whether greater supraclavicular adipose tissue activity could be associated with a healthier metabolic profile in people already diagnosed with type 2 diabetes (T2DM). Thus, the present work evaluated if supraclavicular adipose tissue activity is associated with metabolic and molecular markers in individuals with T2DM. Based on a cluster study, individuals with T2DM were divided into groups according to high or low-standard uptake value (SUV) evaluated in the supraclavicular adipose tissue area by [18F]-fluorodeoxyglucose and positron emission tomography-computed tomography (18F-FDG-PET/CT) after mild cold exposure). Functional, biochemical, inflammatory, and molecular markers were measured. When we evaluated the whole sample, women showed higher SUV, which favored a difference between groups in sex-related markers. On the other hand, volunteers in the high-SUV group showed lower BMI, monocytes count, triglycerides/glucose index (TYG-index) and z score of metabolic syndrome (MS) values, as well as lower triglycerides, and VLDL concentrations. Moreover, they also had enhanced expression of thermogenic genes in subcutaneous fat. When analyzing only women, the differences in markers associated with sex disappear, and a lower count of leukocytes, platelets, along with lower TYG-index, z score of MS values, and triglycerides, VLDL, LDL, and TNFα concentrations were observed in women with the high SUV. In addition, higher expression of thermogenic genes and BECN1 were detected. Higher supraclavicular adipose tissue SUV in individuals with T2DM is associated with a better cardiometabolic/inflammatory profile and expression of thermogenic genes. UTN: U1111-1202-1476 - 08/20/2020.

Oxyresveratrol and/or Dapagliflozin Attenuate Doxorubicin-Induced Nephrotoxicity via Modulation of PPAR-γ/Nrf-2/HO-1, NF-κB/TNF-α/Keap-1, and Bcl-2/Caspase-3/ATG-5 signaling pathways in rats.

Among the most undesirable effects that lead to the restriction of doxorubicin (DOX) use in chemotherapy is kidney damage. This research aimed to assess the possible defenses against DOX-induced nephrotoxicity offered by oxyresveratrol (ORES) and/or dapagliflozin (DAPA). Five groups of eight male Swiss albino rats each were created from a total of sixty-four. One intravenous injection of DOX (10mg/kg) was given into the tail vein on the fourteenth day of the experiment; in the meantime, ORES (80mg/kg) and DAPA (10mg/kg) were given orally 14 days prior to the DOX injection and 2 days following the DOX injection. In rats given DOX, ORES and/or DAPA both successfully reduced the kidney weight, kidney/bodyweight ratio, and blood levels of creatinine, uric acid, and urea. They also increased final body weight and albumin serum levels. Additionally, lower serum concentrations of TNF-α and IL-6 were noted, along with a lower kidney content of caspase-3. Furthermore, the expression of the Bcl-2 gene was upregulated, as were the Nrf-2, PPAR-γ, and HO-1 proteins, and there was a downregulation of the ATG-5, Keap-1, and NF-κB renal gene expression. These findings support a decrease in oxidative stress and relief of histopathological alterations. The current study's findings suggest that ORES and/or DAPA pretreatment could be a viable therapeutic approach to ameliorate DOX-induced nephrotoxicity.

Differential modulation of inflammatory cytokines by recombinant IL-10 in IL-1β and TNF-α ̶ stimulated equine chondrocytes and synoviocytes: impact of washing and timing on cytokine responses

Osteoarthritis (OA) remains a challenging joint disorder necessitating effective anti-inflammatory interventions. In this study, our primary objective was to establish an in vitro protocol that replicates the clinical investigation of anti-inflammatory drugs intended for OA management. Focusing on recombinant IL-10 (r.IL-10) as a potential anti-inflammatory treatment, we designed and implemented two distinct protocols to evaluate the efficacy of r.IL-10 in modulating chondrocyte and synoviocyte inflammation.The experimental design involved sequential stimulation with IL-1β and TNF-α for 24 h, followed by washing (model 1) or not washing (model 2) the cells before r.IL-10 treatment. Samples were collected after 6–24 h of treatment. Cellular responses were evaluated by quantifying gene expression and synthesis of key inflammatory cytokines and proteases.The expression and synthesis of inflammatory cytokines and proteases was significantly affected by washing and treatment time. The expression of IL-1β, TNF-α, IL-8, MMP-13, and ADAMTS5 were effectively reduced in r.IL-10-treated chondrocytes and synoviocytes in model 2 after 24 h, particularly at concentrations of 10 and 20 ng/mL. r.IL-10 treatment significantly increased IL-6 gene expression in chondrocytes at all time points. However, in synoviocytes, IL-6 expression was significantly lower in model 2 after 24 h of r.IL-10 treatment. r.IL-10 treatment significantly decreased IL-1β and TNF-α content in synoviocyte supernatants, particularly in model 2 at concentrations of 10 and 20 ng/mL after 6 and 24 h. r.IL-10 treatment in chondrocytes led to a significant decrease in IL-1β supernatant concentrations in model 2 after 24 h only.This study demonstrated that r.IL-10 treatment effectively reduces key inflammatory markers and matrix metalloproteinase activity in both chondrocytes and synoviocytes, particularly in model 2 where cells were not washed prior to treatment. These findings highlight r.IL-10’s potential as a robust anti-inflammatory agent for OA management and suggest its critical role in developing effective therapeutic strategies for OA.

Probiotic Bacillus cereus regulates metabolic disorders and activates the cholic acid-FXR axis to alleviate DSS-induced colitis

Inflammatory bowel disease is characterized by severe imbalance of intestinal flora and metabolic disorders. Recent studies have demonstrated that probiotics can effectively alleviate inflammatory bowel disease by restoring the intestinal flora structure and modulating the immune response. However, the role of probiotics in regulating intestinal metabolism disorders is still unclear. This study explores the role of probiotic B. cereus in alleviating DSS-induced colitis. The findings indicated probiotic B. cereus treatment mitigated tissue damage and apoptosis during inflammation. Metabolome and transcriptome analysis revealed B. cereus activated the cholic acid-FXR axis by increasing cholic acid levels, which promoted the gene expression level of NF-κB inhibitor α, reduced the IL-1β, IL-6, IL-18 and TNF-α concentrations. Furthermore, it effectively mitigated the DSS-induced disruption of bile acid metabolism, arginine metabolism, and linoleic acid metabolism. This study explores the effect and mechanisms of probiotic B. cereus on alleviating DSS-induced colitis. It aims to provide a theoretical basis for microbial therapy in inflammatory bowel disease. SignificanceThis study used metabolome and transcriptome to reveal the roles and mechanisms, which probiotic Bacillus cereus modulates metabolic disorders and alleviate DSS-induced colitis. We identified the cholic acid-FXR axis as an important target for alleviating DSS-induced colitis. These findings provide new insights into microbial treatment strategies for IBD.

The Role of Chitinase 3-Like-1 (YKL-40) and Proinflammatory Biomarkers in the Pathogenesis of Pediatric Tick-Borne Encephalitis in a Polish Cohort.

Chitinase 3-like-1 (CHI3L1), also known as YKL-40, is a potential biomarker for neuroinflammatory conditions. It is upregulated in Alzheimer's disease, multiple sclerosis, and traumatic brain injury. However, its involvement in pediatric tick-borne encephalitis (TBE) has not been addressed yet. This study aimed to evaluate CHI3L1 and its relationship with other inflammatory cytokines, blood-brain barrier (BBB) integrity, immune response, and disease severity in pediatric patients with TBE. A total of 22 pediatric TBE patients hospitalized in Bialystok, Poland were included in this study. Participants were categorized as having meningoencephalitis (n=6) or meningitis (n=16). The integrity of the brain-blood barrier (BBB) was assessed using the albumin quotient (albQ). Biomarker indices were calculated to account for variations in BBB permeability. The concentrations of CHI3L1, CCL2, chemerin, CXCL2, IFN-γ, IL-1-β, IL-4, IL-6, IL-13, and TNF-α in both serum and CSF, were measured using the Luminex Multiplex Assay od admission and two weeks later when symptoms resolved. CSF and serum concentrations of CHI3L1 did not differ between the encephalitis and meningitis cases. After adjusting for BBB permeability, the CHI3L1 index was 2.4-fold lower in patients with encephalitis than in those with meningitis (P=0.008). There was a post-treatment reduction of CHI3L1, IL-6, and TNF-α CSF concentrations. We also found and improvement in BBB permeability in younger children but in older albQ remained abnormal. Correlation analysis revealed associations between CHI3L1 levels and pro-inflammatory markers, notably chemerin, IL-6, and TNF-α, across both clinical groups. Our findings suggest that CHI3L1 CSF levels reflect the inflammatory activity in pediatric TBE and may help to differentiate between meningoencephalitis and meningitis. The observed interactions between CHI3L1 and other cytokines underscore its potential involvement in inflammatory response to the virus. The prolonged disruption in BBB integrity in older children might reflect age-dependent differences in the severity of TBE. These insights advance our understanding of TBE pathogenesis in children and support further investigation of CHI3L1 as a biomarker for TBE diagnosis and management.

The impact of N-acetylcysteine on lactate, biomarkers of oxidative stress, immune response, and muscle damage: A systematic review and meta-analysis.

N-acetylcysteine (NAC) is a compound whose mechanism of action is intricately linked to the provision of cysteine for glutathione synthesis. It has been used in medicine and has also made significant inroads into sports, as it can modify the levels of several biomarkers, including those of oxidative processes, inflammation and muscle damage after exercise. Because the effectiveness of NAC supplementation is unclear, the primary objective of the present study was to perform a meta-analysis elucidating how NAC supplementation alters the concentrations of GSH (glutathione), GSSG (glutathione disulfide), TBARS (thiobarbituric acid reactive substances), IL-6 (interleukin 6), TNF-α (tumour necrosis factor alpha), CK (creatine kinase), lactate, and muscle soreness after physical exertion. Suitable studies were searched for from February to September 2023, and the results of those included (n = 20) indicate that NAC supplementation significantly diminishes both muscle soreness (p = 0.03; the mean difference (MD) of NAC's effect was -0.43 with a 95% confidence interval (CI), -0.81, -0.04) and lactate concentrations after exercise (p = 0.03; the MD -0.56 mmol/L; 95% CI, -1.07, -0.06). A substantial decrease was observed in concentrations of IL-6 (p = 0.03; the standardized MD (SMD) was -1.71; 95% CI, -3.26, -0.16) and TBARS (p = 0.02; SMD was -1.03, 95% CI, -1.90, -0.15). Furthermore, an elevation in GSH concentration was observed following supplementation. However, we saw no significant effect of NAC on TNF-α, CK or GSSG concentrations. NAC supplementation holds promise for attenuating muscle soreness, lactate, TBARS and IL-6 concentrations and increasing GSH level following physical exertion.

Exploring the Effect and Mechanism of DaYuan Yin Against Acute Lung Injury by Network Pharmacology, Molecular Docking, and Experimental Validation.

DaYuan Yin (DYY), a traditional Chinese medicine for lung diseases, requires further study to understand how it improves acute lung injury (ALI). This study seeks to elucidate the material basis and molecular mechanisms underlying the treatment of ALI with DYY through network pharmacology, molecular docking, and experimental validation. DYY's active components and targets were identified using TCMSP and UHPLC-MS/MS, and a herb-component-target network was created with Cytoscape 3.7.2. ALI target genes were sourced from GeneCards, DisGeNET, and DrugBank. A PPI network was built, with core targets analyzed through GO and KEGG enrichment via Metscape. The therapeutic effects and mechanisms of DYY on LPS-induced ALI in rats were explored, and molecular docking evaluated the interactions between Nrf2, HO-1, TLR4, and the components. The study identified 95 active compounds, 234 therapeutic targets, and 2529 ALI-related genes, with 111 shared targets between DYY and ALI. KEGG analysis indicates that the PI3K-AKT, MAPK, and oxidative stress pathways are associated with DYY's anti-ALI effects. Network pharmacology and UHPLC-MS/MS analysis revealed active ingredients like quercetin, Magnolol, and Wogonin. Compared with the model group, DYY reduced the lung dry-wet ratio (W/D) of ALI rats from (5.31 ± 0.51) to (4.47 ± 0.73)(P < 0.05). Meanwhile, the contents of IL-6 and TNF-α in bronchoalveolar lavage fluid (BALF) and MDA, NO and ROS in lung tissue were also significantly decreased. Notably, DYY enhances UCP2 mRNA expression, boosts Nrf2 and HO-1 expression, and inhibits TLR4-mediated pro-inflammatory mediators. Molecular docking analysis showed that the main components of DYY had strong binding ability with HO-1. DYY can alleviate inflammation, oxidative stress, and ALI-related changes by targeting the Nrf2/HO-1 mediated TLR4 pathway, providing insights for developing effective ALI treatments.

1,25-dihydroxyvitamin D3 regulates enteroglial bioactivity through butyric acid pathway in a high-fat diet mouse model.

1,25-dihydroxyvitamin D3 (1,25(OH)2D3), affects enteric glial cells (EGCs) activity, but the mechanism is still unknown. The current study aimed to explore whether 1,25(OH)2D3 could regulate EGCs activity via butyrate pathway in a high-fat diet model. Male C57BL/6J mice were fed with standard diet (SDD), or vitamin-D-deficient diet (VDD), or high-fat diet (HFD), or HFD plus sodium butyrate (SBR), or HFD plus 1,25(OH)2D3, or HFD plus S100B inhibitor ONO-2506 in vivo. CRL-2690 and Caco-2 cells were treated with palmitic acid (PA) and oleic acid (OA) complex, or S100B, or S100B plus butyric acid (BA) in vitro. 25(OH)D3, 1,25(OH)2D3, TNF-α and S100B concentrations were assayed by enzyme-linked immuno- sorbent assay (ELISA). Colonic mucosal permeability was measured by using FITC-dextran 4kDa. Colonic butyrate was detected using high-performance liquid chromatography (HPLC). The results showed HFD decreased serum 25(OH)D3 and 1,25(OH)2D3 concentrations and colonic butyrate generation. 1,25(OH)2D3 supplementation raised butyrate production in the colon. 1,25(OH)2D3 and sodium butyrate supplementation inhibited EGCs to produce S100B and reduced colonic permeability to FITC-dextran. Inhibition of S100B pathway by ONO- 2506 decreased colonic hyperpermeability. In vitro experiments showed butyrate treatment not only reduced S100B and TNF-α secretion from PA/OA-treated CRL-2690 cells, but also decreased the permeability of S100B-treated Caco-2 cells. Collectively, 1,25(OH)2D3 elicited butyrate to suppress EGCs activation, which helped to prevent intestinal barrier injury.

Preparation of combined colchicine with loxoprofen sodium loaded in dissolvable microneedles and its anti-gouty arthritis effect

Both colchicine (Col) and loxoprofen sodium (LS) are first-line drugs for gouty arthritis (GA). Dissolvable microneedles (DMNs) represent an innovative transdermal drug delivery system (TDDS) that maximizes the therapeutic effect of drugs. In this paper, a novel TDDS of combined Col with LS loaded in DMNs (Col-LS-DMNS) was prepared to treat GA. To assess the synergistic effects of Col and LS, cell viability assays were conducted using monosodium urate (MSU)-induced HaCaT cells. The optimization of the Col-LS-DMNs formulation was carried out through sealing membrane puncture tests, single-factor experiments, and Box-Behnken response surface methodology. The quality of Col-LS-DMNs was assessed using high-performance liquid chromatography (HPLC), scanning electron microscopy (SEM), sealing membrane puncture tests, in vitro mouse skin puncture experiments, and Franz diffusion cell assays. The therapeutic effect of Col-LS DMNs on GA through animal experiments. Results demonstrated that Col and LS exhibit a synergistic effect, with optimal mass ratio of 1:1. The optimal formulation included PVP K30 and CMC Na at a mass ratio of 10:1, with 63% water content and a mixed dosage of Col and LS of 42 mg. The drug content in Col-LS-DMNs was (19.11±0.65) μg of Col and (20.69±0.89) μg of LS per tablet. SEM imaging revealed that the needle tips were conical. Additionally, Col-LS-DMNs exhibited favorable mechanical properties and caused minimal skin damage. After 24 hours, the cumulative permeation amount was (21.55±19.49) μg/cm2. Col-LS-DMNs significantly reduced MSU-induced ankle swelling and decreased serum concentrations of TNF-α, IL-6, and IL-1β in rats. In conclusion, Col-LS-DMNs represent a promising new treatment modality for GA.

Clinical Study of Psycho-cardiology Combined with FNS in the Treatment of SCAD with Depression

To investigate the effects of psycho-cardiology combined with fastigial nucleus stimulation (FNS) on patients with stable coronary artery disease (SCAD) complicated with depression, we observed the changes of concentrations of N-terminal pro-B type natriuretic peptide (NT-proBNP), tumor necrosis factor-α(TNF-α), 5-hydroxytryptamine (5-HT), dopamine (DA) and depressive symptoms in these patients. One hundred and fifty patients, who were hospitalized the Department of Cardiology at the Third People&amp;apos;s Hospital of Mianyang from February 2020 to October 2021 with a confirmed diagnosis of SCAD combined with depression, were selected as study subjects. Patients were randomly divided into a control group, a psycho-cardiology treatment group and a combined treatment group. The control group was given conventional treatment of SCAD and pseudo-stimulation of the fastigial nucleus, the psycho-cardiology treatment group was given conventional treatment of SCAD, psychological interventions and pseudo-stimulation of the fastigial nucleus, and the combined treatment group was given conventional treatment of SCAD, psychological intervention and FNS. The changes of concentrations of TNF-α, NT-proBNP, 5-HT and DA were observed in the three groups of patients, and the changes of the state of depression were assessed by Hamilton Depression Scale (HAMD). Serum NT-proBNP and TNF-α concentrations decreased significantly after treatment in the three groups. Especially, the decrease in the combined treatment group was the most significant(&amp;lt;i&amp;gt;p&amp;lt;0.05&amp;lt;/i&amp;gt;). Serum 5-HT and DA concentrations were not statistically different in the three groups before and after treatment (&amp;lt;i&amp;gt;p&amp;gt;0.05&amp;lt;/i&amp;gt;). However, they were significantly higher in the psycho-cardiology treatment group and the combined treatment group(&amp;lt;i&amp;gt;p&amp;lt;0.05&amp;lt;/i&amp;gt;) and the increase in the combined treatment group was more significant than in the psycho-cardiology treatment group. In the control group, the difference of HAMD scores was not statistically significant before and after treatment (&amp;lt;i&amp;gt;p&amp;gt;0.05&amp;lt;/i&amp;gt;), in the psycho-cardiology treatment group and the combined treatment group, the scores decreased significantly after treatment compared to before treatment (&amp;lt;i&amp;gt;p&amp;lt;0.05&amp;lt;/i&amp;gt;). Furthermore, compared to the psycho-cardiology treatment group, the decrease was more significant in the combined treatment group (&amp;lt;i&amp;gt;p&amp;lt;0.05&amp;lt;/i&amp;gt;). Our data suggested that Psycho-cardiology medicine combined with FNS can effectively reduce the level of serum NT-proBNP and TNF-α, increase the serum concentrations of 5-HT and DA and improve the symptoms of depression in patients with SCAD complicated with depression.

Atractylenolide III ameliorates DSS-induced colitis by improving intestinal epithelial barrier via suppressing the NF-κB-Mediated MLCK-pMLC signaling pathway

This study is to demonstrate the protection of atractylenolide III (AT III) on intestinal barrier dysfunction in ulcerative colitis (UC). UC model was established by 3% dextran sulfate sodium (DSS), and TNF-α was used to induce dysfunction in the intestinal epithelial barrier. TEER, FD-4 transmembrane flux and DAI were measured. Histopathological changes was identified by H&E staining, TJ structure changes were observed by TEM, IL-1β and TNF-α contents were measured by ELISA, bacterial translocation was investigated by FISH. The expressions of ZO-1, occludin, and the proterins in the MLCK/p-MLC and NF-κB pathways were analyzed by Western blotting or immunofluorescence. The results indicated that AT III alleviate the symptoms of DSS-induced colitis, reduce the disruption of intestinal epithelial barrier, and decrease FD4. Moreover, AT III inhibited the destruction of intestinal epithelial TJ structure and bacterial translocation in UC mice. AT III reversed the high levels of IL-1β and TNF-α, the decrease of occludin, ZO-1 expressions. Furthermore, AT III showed similar effects to PDTC (pyrrolidinedithiocarbamate) in ameliorating the disruption of the TNF-α-induced TEER and FD-4 disruption, MLCK protein expression, and MLC2 phosphorylation. In conclusion, AT III mitigates the dysfunction of intestinal epithelial barrier in UC through the NF-κB-mediated MLCK/p-MLC signaling pathway.

Prognostic Value of Baseline Serum Pro-Inflammatory Cytokines in Severe Multisystem Inflammatory Syndrome in Children.

Background: Severe clinical manifestations of multisystem inflammatory syndrome in children (MIS-C) are associated with the dysregulation of immune response following SARS-CoV-2 infection. Therefore, we analyzed the levels of 10 selected cytokines at admission to estimate disease severity and to predict the length of hospitalization. In remission samples, these mediators were followed after intravenous immunoglobulin (IVIG) treatment before discharge. Methods: Thirty-five MIS-C patients at the age of 8.4 ± 4.1 years and 11 clinical controls were included. Acute MIS-C patients were divided into two severity subgroups based on their clinical score determined by the WHO criteria. Serum concentrations of IFN-γ, IL-1α, IL-1RA, IL-8, IL-10, IL-17A, IL-18, IP-10, MCP-1, and TNF-α were measured by MILLIPLEX® Human Cytokine/Chemokine panel, while ACE2 activity was determined by a fluorescent kinetic assay. These results were correlated with routinely determined laboratory parameters and clinical characteristics. Results: MIS-C patients demonstrated significantly elevated baseline levels of most of these cytokines compared to controls. Even higher concentrations of IL-18, TNF-α and ferritin with reduced lymphocyte count were found in severe subjects with elevated clinical scores of 4-5 compared to moderate cases with a clinical score of 1-3. Furthermore, the development of cardiovascular dysfunction and prolonged hospitalization (≥8 days) were related to augmented ACE2 and IL-6 levels. IL-18, IL-1RA, IL-10 and TNF-α were diminished in response to IVIG treatment in remission samples. Finally, pre-treatment IL-18 (≥516.8 pg/mL) and TNF-α (≥74.2 pg/mL) effectively differentiated disease severity in MIS-C with AUC values of 0.770 and 0.750, respectively. Conclusions: IL-18 and TNF-α have a prognostic value in disease severity at admission and are capable of monitoring the efficacy of IVIG treatment in MIS-C.

Acupoint injection inhibiting abnormal secretion of mucin and inflammatory reaction of nasal mucosa in rats with allergic rhinitis through the p38 mitogen-activated protein kinase pathway

To observe the effects of acupoint injection on the expression of p38 mitogen-activated protein kinase (MAPK), phosphorylated (p)-p38 MAPK, mucin 5 subtype AC (MUC5AC) in the nasal mucosa, and serum inflammatory factors of rats with allergic rhinitis, so as to explore the mechanism of acupoint injection in improving inflammatory reactions in the nasal mucosa of rats with allergic rhinitis. SD rats were randomly divided into the normal group, model group, acupoint injection group, and non-acupoint group, with 7 rats in each group. The allergic rhinitis rat model was established using ovalbumin sensitization. Intervention was performed by injecting dexamethasone and 1% lidocaine mixture (0.05 mL) into the bilateral "Yingxiang"(LI20) and "Yintang"(GV24+) acupoints, with injection at non-acupoints for non-acupoint group as the control, once every 3 days for a total of 4 times. Allergic symptoms in the rat nose were evaluated using a cumulative scoring method；histopathological changes in the nasal mucosa were observed after HE staining；serum histamine (HA), interleukin (IL)-6, IL-8, and tumor necrosis factor-alpha (TNF-α) contents were detected using ELISA；real-time fluorescent quantitative PCR was used to detect the expression of MUC5AC mRNA in the nasal mucosa；Western blot was used to detect the expression of p38 MAPK, p-p38 MAPK, and MUC5AC protein in the nasal mucosa. Compared with the normal group, nasal allergic symptom score, serum HA, IL-6, IL-8, and TNF-α contents, nasal mucosal p38 MAPK, p-p38 MAPK proteins expression, MUC5AC mRNA and protein expression were significantly increased(P<0.01) in the model group. Compared with the model group and non-acupoint group, nasal allergic symptom score, serum HA, IL-6, IL-8, and TNF-α contents, nasal mucosal p38 MAPK and p-p38 MAPK proteins expression, MUC5AC mRNA and protein expression were significantly reduced (P<0.01, P<0.05) in the acupoint injection group. Acupoint injection can reduce nasal allergic symptom, and MUC5AC secretion in rats with allergic rhinitis, alleviate nasal mucosal inflammatory reactions, and its mechanism of action may be related to the inhibition of p38 MAPK phosphorylation, thereby reducing the release of inflammatory factors.

Effects of electroacupuncture on expression of miR-218 and HMGB1/TLR4 signaling pathway in rats with focal cerebral ischemia

To observe the effect of electroacupuncture (EA) on high mobility group box 1 (HMGB1) / toll-like receptor 4 (TLR4) signaling pathway and inflammatory injury in rats with focal cerebral ischemia (FCI), so as to explore its mechanisms underlying amelioration of ischemic stroke. Male SD rats were randomly divided into sham operation (n=18), model (n=18) and EA (n=18) groups. The FCI model was established according to Longa's method. In the EA group, 24 h after modeling, EA stimulation was applied to "Baihui"(GV20), "Fengfu"(GV16), and "Neiguan" (PC6) and "Xinshu" (BL15) on the left side for 20 min, once a day for 1 week. The neurological deficit severity was evaluated by the modified neurological severity score (mNSS), and the morphological changes of neurons were observed after H.E. staining. The expression of HMGB1 and TLR4 proteins were detected by immunofluorescence staining. The contents of tumor necrosis factor(TNF)-α and interleukin(IL)-6 in the ischemic area tissue of the brain were detected using ELISA. The mRNA expressions of HMGB1 and TLR4 as well as the expression of miR-218 in the ischemic cortex were detected by PCR. Compared with the sham operation group, the mNSS at 2 h, 48 h and 7 day (d), IL-6 and TNF-α contents, HMGB1 and TLR4 immunoactivity and mRNA expression at 48 h and 7 d were significantly increased (P<0.01), while miR-218 expression at 48 h and 7 d was obviously down-regulated (P<0.01) in the model group. In comparison with the model group, the mNSS at 7 d, IL-6 and TNF-α contents, HMGB1 and TLR4 immunoactivity and mRNA expression at 7 d not at 48 h were significantly decreased (P<0.01, P<0.05), while the expression of miR-218 at 7 d was apparently up-regulated (P<0.05) in the EA group. H.E. staining showed that there were signs of neuronal necrosis including blurred outline of the cells, disordered internal structure, dense cytoplasm, shrunk nuclei, etc. in the ischemic area of the brain at 48 h and 7 d in the model group, which was evidently milder on day 7 in the EA group. EA can reduce the inflammatory injury of brain tissue and improve the neurological impairment in rats with FCI, which may be associated with its effects in inhibiting HMGB1/TLR4 signaling and down-regulating the level of pro-inflammatory cytokines.

Molecular Changes in Aqueous Humor Associated with Inflammation Following Cataract Surgery in Patients with Fuchs' Endothelial Corneal Dystrophy.

To evaluate the anterior chamber (AC) inflammation in the early postoperative period after cataract surgery and before Descemet membrane endothelial keratoplasty (DMEK) by quantifying oxidative stress and inflammatory mediators in aqueous humor of patients with Fuchs' endothelial corneal dystrophy (FECD). In this prospective single-center study, 15 patients with FECD underwent cataract surgery and DMEK in a two-stage procedure.Aqueous humor was collected from the AC at the beginning of cataract surgery and 3months later at the beginning of DMEK. In the control group, which consisted of 15 age-matched phakic patients without FECD, aqueous humor was only collected at the beginning of cataract surgery. Mediators of postoperative inflammation including TNF-α, VEGF, IL-2, IL-1 β, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12, GM-CSF, IFN-γ, CXCL5/ENA-78, FGF-basic, G-CSF, IL-1-α, IL-1-ra, IL-17, CCL2/MCP-1, CCL3/MIP-1a, CCL4/MIP-1b, TPO, TGF-β-1, TGF-β-2, and TGF-β-3 concentrations were measured using a Multiplex-Array-System. The concentration of TNF-α (p = 0.021), IL-6 (p = 0.005), IL-8 (p = 0.001), CXCL5/ENA78 (p = 0.002), CCL2/MCP-1 (p = 0.001) and CCL4/MIP-1b (p = 0.037) were significantly higher 3months after cataract surgery at the beginning of DMEK compared to control group at beginning of cataract surgery. The levels of IL-2, IL-5, IL-8, IL-10, and IL-1-α were significantly higher in phakic eyes in the control group (p < 0.05) before cataract surgery. The present study indicates significantly increased proinflammatory cytokines 3months after cataract surgery in eyes with FECD. Our findings suggest postoperative inflammation in the AC up to 3months after cataract surgery. Therefore, it may be reasonable to combine cataract surgery with DMEK in cataract patients with FECD.