Serum Levels Of Proinflammatory Cytokines Research Articles

Interleukin-33 Ameliorates Murine Systemic Lupus Erythematosus and Is Associated with Induction of M2 Macrophage Polarisation and Regulatory T Cells

The innate cytokine IL-33 is increasingly recognised to possess biological effects on various immune cells. We have previously demonstrated elevated serum level of soluble ST2 in patients with active systemic lupus erythematosus suggesting involvement of IL-33 and its receptor in the lupus pathogenesis. This study sought to examine the effect of exogenous IL-33 on disease activity of pre-disease lupus-prone mice and the underlying cellular mechanisms. Recombinant IL-33 was administered to MRL/lpr mice for 6 weeks, whereas control group received phosphate-buffered saline. IL-33-treated mice displayed less proteinuria, renal histological inflammatory changes, and had lower serum levels of pro-inflammatory cytokines including IL-6 and TNF-α. Renal tissue and splenic CD11b+ extracts showed features of M2 polarisation with elevated mRNA expression of Arg1, FIZZI, and reduced iNOS. These mice also had increased IL-13, ST2, Gata3, and Foxp3 mRNA expression in renal and splenic tissues. Kidneys of these mice displayed less CD11b+ infiltration, had downregulated MCP-1, and increased infiltration of Foxp3-expressing cells. Splenic CD4+ T cells showed increased ST2-expressing CD4+Foxp3+ population and reduced IFN-γ+ population. There were no differences in serum anti-dsDNA antibodies and renal C3 and IgG2a deposit in these mice. Exogenous IL-33 was found to ameliorate disease activity in lupus-prone mice with induction of M2 polarisation, Th2 response, and expansion of regulatory T cells. IL-33 likely orchestrated autoregulation of these cells through upregulation of ST2 expression.

Bone marrow stem cells incubated with ellipticine regenerate articular cartilage by attenuating inflammation and cartilage degradation in rabbit model.

Ellipticine (Ellip.) was recently reported to have beneficial effects on the differentiation of adipose-derived stem cells into mature chondrocyte-like cells. On the other hand, no practical results have been derived from the transplantation of bone marrow stem cells (BMSCs) in a rabbit osteoarthritis (OA) model. This study examined whether autologous BMSCs incubated with ellipticine (Ellip.+BMSCs) could regenerate articular cartilage in rabbit OA, a model similar to degenerative arthritis in human beings. A portion of rabbit articular cartilage was surgically removed, and Ellip.+BMSCs were transplanted into the lesion area. After two and four weeks of treatment, the serum levels of proinflammatory cytokines, i.e., tumor necrosis factor α (TNF-α) and prostaglandin E2 (PGE2), were analyzed, while macroscopic and micro-computed tomography (CT) evaluations were conducted to determine the intensity of cartilage degeneration. Furthermore, immuno-blotting was performed to evaluate the mitogen-activated protein kinases, PI3K/Akt, and nuclear factor-κB (NF-κB) signaling in rabbit OA models. Histological staining was used to confirm the change in the pattern of collagen and proteoglycan in the articular cartilage matrix. The transplantation of Ellip.+BMSCs elicited a chondroprotective effect by reducing the inflammatory factors (TNF-α, PGE2) in a time-dependent manner. Macroscopic observations, micro-CT, and histological staining revealed articular cartilage regeneration with the downregulation of matrix-metallo proteinases (MMPs), preventing articular cartilage degradation. Furthermore, histological observations confirmed a significant boost in the production of chondrocytes, collagen, and proteoglycan compared to the control group. Western blotting data revealed the downregulation of the p38, PI3K-Akt, and NF-κB inflammatory pathways to attenuate inflammation. The transplantation of Ellip.+BMSCs normalized the OA condition by boosting the recovery of degenerated articular cartilage and inhibiting the catabolic signaling pathway.

Lactoferrin alleviates Western diet-induced cognitive impairment through the microbiome-gut-brain axis

Lactoferrin (Lf) has been shown to benefit cognitive function in several animal models. To elucidate the underlying mechanisms, male C57BL/6J mice were randomly divided into the control (CON), Western-style diets (WD), lactoferrin (Lf), and Lf + antibiotics (AB) groups. The Lf group was intragastrically administered with Lf, and the Lf + AB group additionally drank a solution with antibiotics. After 16 weeks of intervention, Lf improved the cognitive function as indicated by behavioral tests. Lf also increased the length and curvature of postsynaptic density and upregulated the related protein expression, suggesting improved hippocampal neurons and synapses. Lf suppressed microglia activation and proliferation as revealed by immunofluorescence analysis. Lf decreased the serum levels of pro-inflammatory cytokines and downregulated their protein expressions in the hippocampus region. Lf also inhibited the activation of NF-κB/NLRP3 inflammasomes in the hippocampus. Meanwhile, Lf upregulated the expression of tight junction proteins, and increased the abundance of Bacteroidetes at phylum and Roseburia at genus, which are beneficial for gut barrier and cognitive function. The antibiotics eliminated the effects of long-term Lf intervention on cognitive impairment in the Lf + AB group, suggesting that gut microbiota participated in Lf action. Short-term Lf intervention (2 weeks) prevented WD-induced gut microbiota alteration without inducing behavioral changes, supporting the timing sequence of gut microbiota to the brain. Thus, Lf intervention alleviated cognitive impairment by inhibiting microglial activation and neuroinflammation through the microbiome-gut-brain axis.

Highlights on molecular targets in the management of COVID-19: Possible role of pharmacogenomics

By the end of 2022, there had been a reduction in new cases and deaths caused by coronavirus disease 2019 (COVID-19). At the same time, new variants of the severe acute respiratory syndrome coronavirus 2 virus were being discovered. Critically ill patients with COVID-19 have been found to have high serum levels of proinflammatory cytokines, especially interleukin (IL)-6. COVID-19-related mortality has been attributed in most cases to the cytokine storm caused by increased levels of inflammatory cytokines. Dexamethasone in low doses and immunomodulators such as IL-6 inhibitors are recommended to overcome the cytokine storm. This current narrative review highlights the place of other therapeutic choices such as proteasome inhibitors, protease inhibitors and nuclear factor kappa B inhibitors in the treatment of patients with COVID-19.

PS-R05-3: SEVERE GLOMERULAR MICROANGIOPATHY IN A PATIENT WITH NEPHROTIC SYNDROME FOLLOWING TAKAYASU ARTERITIS

Introduction: Takayasu arteritis (TA) is a large vessel inflammation that predominantly involves aorta and its main arteries. TA-caused kidney injury is known to be mainly due to renal artery stenosis. However, glomerulonephritis including membranous proliferative glomerulonephritis, focal segmental sclerosis, or crescentic nephritis has been histologically identified in some TA cases. Case description: A 69-year-old female presented with leg edema and refractory hypertension with nephrotic range proteinuria (17.0 g/gCr). She exhibited > 10 mmHg blood pressure discrepancy between left and right upper limbs and severe aortic regurgitation. Lab test showed high level of C-reactive protein and high erythrocyte sedimentation rate. Contrast-enhanced computed tomography identified wall thickening of the left subclavian artery, which led to the diagnosis of TA. Renal biopsy demonstrated that double basement membrane, enlarged subendothelial space, and mesangiolysis with no immunoglobulin deposition. Although azilsartan 40 mg QD and nifedipine 30 mg BID had been prescribed, systolic blood pressure (sBP) was still > 160 mmHg. Thus, while oral prednisolone was started to suppress TA-induced inflammation, azilsartan was replaced with sacubitril valsartan, an angiotensin receptor-neprilysin inhibitor (ARNI), for treating refractory hypertension. After a few weeks, sBP was decreased to 130 mmHg with sacubitril valsartan 200 mg QD and proteinuria was dramatically improved to 0.5 g/gCr. Discussion: TA commonly involves the renal artery, leading to stenosis and subsequent ischemic nephropathy. However, no stenosis and occlusion were observed in renal artery of this case. In addition, small vessel inflammation in the kidney such as vessel wall necrosis and immune cells infiltration around arterioles was not found. Serum levels of pro-inflammatory cytokines including vascular endothelial growth factor and interleukin-6, a well-known contributors for progression of TA-related inflammation, were within the normal range. The results indicate that TA-caused hypertension, but not vessel inflammation induced by TA, seems to play a pathological role for development of glomerular microangiopathy in this case. A previous study showed that 70% of TA patients presenting with glomerular lesion has severe hypertension and intimal thickening in renal arterioles was observed more frequently in TA patients presenting with glomerular lesion when compared to those without glomerulonephritis, which supports our hypothesis. In the present case, ARNI significantly reduced sBP which might directly contribute to the improvement of proteinuria. ARNI can be potent to control hypertension more effectively than conventional hypertensive agents in TA patients.

An in-vivo experimental evaluation of the efficacy of fish-derived antimicrobial peptides against multidrug-resistant Pseudomonas aeruginosa.

due to the fact that antimicrobial peptides antimicrobial peptides (AMPs) from climbing perch have not been fully explored for their antimicrobial potency, this investigation was undertaken to explore that possibility. antimicrobial peptides (AMPs) from the mucous secretion of climbing perch were obtained and an in-vivo analysis was conducted using mice. the results showed inhibitory effects on multidrug-resistant multidrug-resistant Pseudomonas aeruginosa with reduced mortality from 100% among the non-treated group to 25%. Similarly, the level of serum transaminase enzymes (AST and ALT), creatinine levels, and pro-inflammatory cytokines (TNF-α and IL-6) were all found to be higher in the non-treatment group compared to the AMP-treatment group. Also, extensive tissue damage in the lung, liver, and spleen of the non-treated control group mice was observed based on the histopathological lesions recorded. As expected, AMPs from climbing perch significantly alleviated multidrug-resistant P. aeruginosa infection in-vivo and produced enhanced therapeutic efficacy superior to the ciprofloxacin treatment. this study provides insight into the potential antimicrobial activity of fish innate immune system-derived peptides that could serve as a candidate for the substitute of antibiotics.

Clinical characteristics and manometric findings of esophageal achalasia—a systematic review regarding differences among three subtypes

Esophageal achalasia is classified into three subtypes according to manometric findings. Since several factors, including clinical characteristics and treatment response, have been reported to differ among the subtypes, the underlying pathogenesis may also differ. However, a comprehensive understanding regarding the differences is still lacking. We therefore performed a systematic review of the differences among the three subtypes of achalasia to clarify the current level of comprehension. In terms of clinical features, type III, which is the least frequently diagnosed of the three subtypes, showed the oldest age and most severe symptoms, such as chest pain. In contrast, type I showed a higher prevalence of lung complications, and type II showed weight loss more frequently than the other types. Histopathologically, type I showed a high loss of ganglion cells in esophagus, and on a molecular basis, type III had elevated serum pro-inflammatory cytokine levels. In addition to peristalsis and the lower esophageal sphincter (LES) function, the upper esophageal sphincter (UES) function of achalasia has attracted attention, as an impaired UES function is associated with severe aspiration pneumonia, a fatal complication of achalasia. Previous studies have indicated that type II shows a higher UES pressure than the other subtypes, while an earlier decline in the UES function has been confirmed in type I. Differences in the treatment response are also crucial for managing achalasia patients. A number of studies have reported better responses in type II cases and less favorable responses in type III cases to pneumatic dilatation. These differences help shed light on the pathogenesis of achalasia and support its clinical management according to the subtype.

Dietary supplementation with Clostridium butyricum and its ferment substance improves the egg quality and ovarian function in laying hens from 50 to 58 weeks of age.

The current study was conducted to explore the effects of dietary Clostridium butyricum (C.butyricum) and fermented calcium (Ca) butyrate produced by C.butyricum on the performance and egg quality of post-peak laying. A total of 384 50-week-old hens were fed a basal diet, the basal diet with 300 mg/kg of fermented Ca butyrate or 1 × 109 CFU/kg C.butyricum for 8weeks. Hens received a C.butyricum exhibited higher yolk properties, albumen height, and Haugh unit. A diet with fermented Ca butyrate or C.butyricum increased the egg mass and the pre-grade yellow follicle number. RNA-sequencing (RNA-seq) data showed that these observations were associated with cytokine-cytokine receptor interaction and intestinal immune status. Accordingly, when compared with the basal diet group, Ca butyrate and C.butyricum addition decreased serum pro-inflammatory cytokine levels and increased the concentration of immunoglobulin A, along with improved intestinal barrier. In addition, dietary C.butyricum inclusion induced a higher abundance of Ruminococcaceae and Lachnospiraceae at the family level. In summary, dietary fermented Ca butyrate or C.butyricum supplementation improved egg quality and ovarian function, which might be related to the enhanced intestinal barrier and immunity in post-peak laying hens.

Molecular and cellular features of femoral head avascular necrosis: in vivo study

Aim. To study the molecular and cellular features of femoral head avascular necrosis in the rat model.Materials and Methods. Femoral head avascular necrosis was surgically induced in 8 rats with the 4-week follow-up. Then, the animals have been euthanised, and we performed gross, radiological, and histological examination of avascular and intact contralateral femoral heads. Systemic inflammation was assessed using enzyme-linked immunosorbent assay, and pro-inflammatory cytokines (interleukin-1β, interleukin-6, and tumor necrosis factor α). The proteomic profile of healthy and necrotic femoral heads was interrogated using sodium dodecyl sulfate polyacrylamide gel electrophoresis and ultra-high performance liquid chromatography-tandem mass spectrometry with ion mobility (TimsToF Pro).Results. Aseptic necrosis of the femoral head was successfully induced in all rats. Serum levels of pro-inflammatory cytokines (interleukin-1β and interleukin-6) were higher in rats with femoral head avascular necrosis as compared with healthy rats. Among the major proteins revealed at proteomic profiling were those involved in maintaining bone tissue homeostasis, calcium phosphate metabolism, angiogenesis, hematopoiesis, cell-cell interactions, chaperones, cartilage matrix proteins, collagen synthesis, and lipid metabolism. In bones with avascular necrosis, we have also found proteins regulating the inflammatory response and oxidative stress. Sodium dodecyl sulfate polyacrylamide gel electrophoresis indicate that the development of avascular osteonecrosis was accompanied by an overexpression of oxidative stress proteins, anaerobic glycolysis, and non-specific inflammatory response along with the downregulation of molecules responsible for angiogenesis, chondrogenesis, calcium phosphate metabolism, collagen synthesis, and cartilage matrix.Conclusion. Femoral head avascular necrosis is accompanied by non-specific inflammation, oxidative stress, and lipid peroxidation all presumably developed because of hypoxia and together contributing to bone destruction.

Exosomal RBP4 potentiated hepatic lipid accumulation and inflammation in high-fat-diet-fed mice by promoting M1 polarization of Kupffer cells

Exosomes containing various biological cargoes have potential to be novel diagnostic biomarkers for metabolic diseases. In this study, retinol-binding protein 4 (RBP4) was found to be enriched in serum exosomes, and its increased levels could be considered as an independent risk factor for the pathogenesis of nonalcoholic fatty liver disease (NAFLD). Exosomal RBP4 (exo-RBP4), primarily derived from hepatocytes, significantly enhanced the M1-like polarization of Kupffer cells (KCs) via promoting the activation of NOX2 and NF-κB and reactive oxygen species (ROS) accumulation, resulting in the over-production of inflammatory cytokines including TNF-α. Subsequently, those excess cytokines remarkably increased the levels of intracellular free fatty acid uptake and lipogenesis-related genes (FAS and SREBP-1c) but decreased fatty acid degradation-related genes (CPT-1 and PPARα) in palmitic acid-treated LO2 cells. More notably, TNF-α significantly elevated RBP4 transcription by activating STAT3 in hepatocytes, playing a positive role in NAFLD development. Intravenous injection with RBP4 (50 μg/kg) potentiated hepatic lipid accumulation, M1-type KC proportion, and serum pro-inflammatory cytokine levels in the hepatic tissues of high-fat-diet-fed mice. Collectively, these data indicated that exo-RBP4 converted KCs to M1 subtype by mediating the NOX2/ROS/NF-κB pathway, subsequently promoting de novo lipogenesis in hepatocytes by TNF-α secretion to activate the JAK2/STAT3 signaling pathway. Therefore, this study uncovered a novel intercellular communication between the inflammatory microenvironment and lipid metabolism for fostering NAFLD progression and found the potential of exo-RBP4 as a novel diagnostic biomarker and therapeutic target for NAFLD.

Higher serum interleukin-12 levels are associated with the pathophysiology of major depressive disorder: A case-control study results.

Major depressive disorder (MDD) is the fourth biggest health-related concern that dramatically impacts individuals' mental and physical health. Alteration of serum proinflammatory cytokine levels may take part in the development and progression of MDD. We aimed to explore and compare the role of interleukin-12 (IL-12) in MDD patients and healthy controls (HCs) and its involvement with the disease severity. The present study included 85 patients and 87 age-sex matched HCs. A qualified psychiatrist utilized the diagnostic and statistical manual of mental disorders, fifth edition (DSM-5) criteria to diagnose patients and evaluate HCs. We applied the Ham-D rating scale to measure the severity of depression. Serum IL-12 levels were measured using ELISA kits. We observed a notable increase in the serum levels of IL-12 in MDD patients compared to HCs (164.27 ± 10.18 pg/ml and 82.55 ± 4.40 pg/ml; p < 0.001). Moreover, we noticed a positive correlation between serum IL-12 levels and Ham-D scores in MDD patients (r = 0.363; p = 0.001). Receiver operating characteristicanalysis showed a good predictive performance (AUC = 0.871; p < 0.001)at the cut-off point of 53.46 pg/ml for serum IL-12. The current study findings support that IL-12 levels are involved with the pathogenesis and inflammatory process in MDD. At the same time, this involvement may make this cytokine eligible for the risk evaluation of MDD. However, we recommend further interventional studies to explore more accurate associations between IL-12 and depressive disorder.

Anti-Osteoarthritic Effects of Cartilage-Derived Extracellular Matrix in a Rat Osteoarthritis Model.

The extracellular matrix (ECM) has many functions, such as segregating tissues, providing support, and regulating intercellular communication. Cartilage-derived ECM (CECM) can be prepared via consecutive processes of chemical decellularization and enzyme treatment. The purpose of this study was to improve and treat osteoarthritis (OA) using porcine knee articular CECM. We assessed the rheological characteristics and pH of CECM solutions. Furthermore, we determined the effects of CECM on cell proliferation and cytotoxicity in the chondrocytes of New Zealand rabbits. The inhibitory effect of CECM on tumor necrosis factor (TNF)-α-induced cellular apoptosis was assessed using New Zealand rabbit chondrocytes and human synoviocytes. Finally, we examined the in vivo effects of CECM on inflammation control and cartilage degradation in an experimental OA-induced rat model. The rat model of OA was established by injecting monosodium iodoacetate into the intra-articular knee joint. The rats were then injected with CECM solution. Inflammation control and cartilage degradation were assessed by measuring the serum levels of proinflammatory cytokines and C-telopeptide of type II collagen and performing a histomorphological analysis. CECM was found to be biocompatible and non-immunogenic, and could improve cell proliferation without inducing a toxic reaction. CECM significantly reduced cellular apoptosis due to TNF-α, significantly improved the survival of cells in inflammatory environments, and exerted anti-inflammatory effects. Our findings suggest that CECM is an appropriate injectable material that mediates OA-induced inflammation.

Caffeic acid protects against atherosclerotic lesions and cognitive decline in ApoE−/− mice

Caffeic acid has been indicated to benefit cholesterol balance, but the effect of pure caffeic acid on atherosclerosis in vivo has not been tested. Given that atherosclerosis and Alzheimer's disease share common features including distracted lipid balance and chronic inflammation, the concurrent effects of caffeic acid on atherosclerotic lesions and cognitive decline were explored here by using the ApoE−/− mice model. A two months' administration of 20 mg/kg caffeic acid or saline was given once two days intraperitoneally to 5-month-old female ApoE−/− mice. We found that the caffeic acid treatment reduced the atherosclerotic lesions in the whole aorta and aortic sinus of the resulting 7-month-old ApoE−/− mice by roughly 50%, compared with the saline control. Meanwhile, the cognitive decline of treated mice were significantly alleviated, as measured by Y-maze and Morris water maze tasks. A reduced accumulation of β-amyloid in the hippocampus was also observed. These effects were associated with elevated serum HDL-c concentration, upregulated ABCA1 and ABCG1 mRNA levels, as well as decrease local inflammation and reduced levels of serum pro-inflammatory cytokines including TNF-α, IL-6 and MCP-1. These obtained results suggested the preventive and therapeutic potential of caffeic acid against atherosclerosis and Alzheimer's disease during aging.

Rice Germ Ameliorated Chronic Unpredictable Mild Stress-Induced Depressive-like Behavior by Reducing Neuroinflammation.

Stress-induced neuroinflammation is widely regarded as one of the primary causes of depression. Gamma-aminobutyric acid (GABA)-enriched foods relieve stress and reduce inflammatory reactions. This study aimed to evaluate whether rice germ with 30% GABA (RG) reduced neuroinflammation in mice exposed to chronic unpredictable mild stress (CUMS). CUMS mice were administered 40, 90, and 140 mg/kg of RG. CUMS increased serum and hypothalamic pro-inflammatory cytokine (TNF-α and IL-6) levels, which were decreased by RG. In the hypothalamus, CUMS elevated M1-type microglia markers of CD86 and NF-κB, whereas RG lowered these levels. The expression levels of NLRP3 inflammasome complex (NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain, and caspase-1), IL-1β, and IL-18 were increased in the hypothalamus of CUMS mice and decreased by RG. RG attenuated depressive-like behaviors in CUMS mice, as measured by the forced swim test and tail suspension test. In conclusion, RG decreased hypothalamic inflammation-related signals, such as TNF-α, IL-6, M1 polarization, NF-κB, NLRP3 inflammasome complex, caspase-1, IL-1β, and IL-18, to diminish depressive-like behavior.

Heat-Killed Bifidobacterium bifidum B1628 May Alleviate Dextran Sulfate Sodium-Induced Colitis in Mice, and the Anti-Inflammatory Effect Is Associated with Gut Microbiota Modulation.

Inflammatory bowel disease (IBD) is a chronic inflammatory disease associated with gut dysbiosis. This study aimed to investigate the effects of heat-killed Bifidobacterium bifidum B1628 (HB1628) in dextran sulfate sodium (DSS)-induced colitis in mice. The following three mouse groups were included (n = eight per group): NC (normal control), DSS (colitis), and HB1628 (colitis and postbiotic). The mice in the DSS group showed significant weight loss and histological damage, developed bloody diarrhea, scored high in the disease activity index (DAI), and exhibited increases in pro-inflammatory cytokines (interleukin [IL]-1β, IL-6, and tumor necrosis factor [TNF]-α) and decreases in an anti-inflammatory cytokine (IL-13) in the serum. These changes were accompanied by gut microbiota modulation in colitis mice (decreases in Rikenellaceae and Eubacterium; increases in Peptostreptococcaceae, Bacteroides vulgatus, and Parasutterella excrementihominis). The HB1628 group had lower DAIs, histology scores, and serum levels of pro-inflammatory cytokines (IL-1β and TNF-α), but higher levels of an anti-inflammatory cytokine (IL-13), compared with the DSS group, suggesting a less severe inflammatory state after the HB1628 intervention. Additionally, HB1628 improved DSS-induced gut dysbiosis, which is evidenced by increases in intestinal beneficial bacteria, such as Lactobacillus, and decreases in known unfavorable taxa in IBD, e.g., Porphyromonadaceae, Subdoligranulum, Lachnospiraceae bacterium 3_1_46FAA, and Alistipes indistinctus. Functional metagenomics revealed three significantly enriched metabolic pathways in the HB1628 group (namely, the aerobic respiration I [cytochrome c] pathway and the superpathways of L-phenylalanine biosynthesis and L-tryptophan biosynthesis, respectively). In conclusion, our results showed that HB1628 effectively improved the inflammation state and tissue damage in DSS-induced colitis mice, and the symptom relief effect was accompanied by obvious gut microbiota remodulation.

Associations of Clonal Hematopoiesis with Recurrent Vascular Events and Death in Patients with Incident Ischemic Stroke

Clonal hematopoiesis (CH), defined by the presence of somatic mutations in hematopoietic stem cells of individuals without hematologic disease, is common among older people and associated with an increased risk of atherosclerosis, inflammation, and shorter overall survival. The mechanism leading to cardiovascular disease is best understood for TET2, one of the most frequently mutated genes, for which accelerated development of atherosclerosis driven by an altered function of the NLRP3/IL-1β-inflammasome in mutated macrophages was reported in preclinical models. CH has recently been associated with a higher risk of stroke in large patient series, however its role with respect to secondary vascular risk and death after ischemic stroke remains unknown. To fill this gap, we investigated CH in peripheral blood DNA from 585 patients with first-ever ischemic stroke from the Prospective Cohort with Incident Stroke-Berlin study (PROSCIS-B) using error-corrected targeted sequencing of 45 genes recurrently mutated in CH. The primary composite endpoint (CEP) consisted of recurrent stroke, myocardial infarction, and all-cause mortality. We identified 355 somatic mutations with a variant allele frequency (VAF) ≥ 1% in 240/585 patients (41%). While 158 patients harbored a single mutation, 82 patients harbored multiple (up to five) mutations. The most frequently mutated genes were DNMT3A, TET2, ASXL1 and PPM1D. In terms of the different etiological subtypes of ischemic stroke, CH was associated with large-artery atherosclerosis stroke (odds ratio 1.60, 95%-CI 1.07 - 2.44, P = 0.024) in logistic regression after correcting for age, sex and cardiovascular risk factors. White matter lesion load as quantified by the Wahlund score was significantly higher in CH-positive patients in baseline MRI compared to CH-negative patients (median Wahlund score 7, IQR 4 - 11 vs. 4, IQR 2 - 8, P < 0.001). To determine whether CH was associated with systemic inflammation, we measured serum levels of pro-inflammatory cytokines shortly after stroke. CH-positive patients showed increased serum levels of IL-6, hsCRP, and VCAM-1 as compared to CH-negative patients (Fig. 1), supporting the current concept that CH mutations alter the inflammatory properties of myeloid cells. With a median follow-up time of 36.2 months, primary endpoint data were available for all 585 patients with a total of 98 events. In univariable Kaplan-Meier analysis, CH was associated with a higher risk for recurrent vascular events and death (hazard ratio [HR] 1.55, 95%-CI 1.05 - 2.31, P = 0.02), which was more pronounced in patients with larger clones (Fig. 2). This led us to consider CH clone size as estimated by the VAF as a continuous parameter with respect to the primary endpoint. In fact, clone size remained an independent risk factor for the CEP in multivariable Cox regression (HR 1.21, 95%-CI 1.00 - 1.46, P = 0.046) after adjustment for age, sex, stroke severity, cardiovascular risk factors, atrial fibrillation, and peripheral artery disease. On single gene level, this association was mainly driven by TET2-mutated CH (HR 2.18, 95%-CI 1.11 - 4.26, P = 0.023) and PPM1D-mutated CH (HR 7.00, 95%-CI 2.27 - 21.73, P < 0.001). Our data indicate that CH is associated with large-artery atherosclerosis and an increased risk of recurrent vascular events and death in patients with ischemic stroke, in particular in patients with larger, and TET2- or PPM1D-mutated clones. The CH mutation profile is accompanied by a systemic pro-inflammatory profile, potentially opening new avenues for preventive precision medicine approaches to resolve the self-perpetuating cycle of inflammation and clonal expansion. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

The level pro-inflammatory and anti-inflammatory biomarkers in patients with chronic mechanical low back pain under pulse radiofrequency therapy

Low back pain is a frequent and recurrent condition, often with a non-specific cause. Conventional treatment methods are generally insufficient in the treatment of chronic low back pain. The aim of the study was to estimate the level of IFN, IL-1, IL-6 (proinflammatory), IL-10, IL-4 (anti-inflammatory) and VEGF proteins in the serum of patients with chronic mechanical low back pain under Pulse radiofrequency (PRF) therapy. The study was carried out on 40 patients 20-60 years old, diagnosed with chronic low back pain for at least 4 months, primary complaint on lumbosacral low back pain, pain intensity VAS (visual analog scale) score of 5 and above, not responding well to conservative treatment (analgesic drugs, physiotherapy, etc.). Therapeutic Radiofrequency applications were carried out with an RF generator (RFG 3C Plus, Radionics). Blood samples were taken 1 day before interventional treatment (control), then 1 day (group1) and 15 days (group 2) after. The serum level of IFN, IL-1, IL-6, IL-10, IL-4 and VEGF l was analyzed with ELISA test. It was shown that as a result of PRF treatment the level of IL-1 was decreased while the levels of IL-4 and IL-6 were increased. It was concluded that the increase in serum levels of proinflammatory cytokines may be correlated with the severity of pain and that the increase in the level of anti-inflammatory cytokines reduces pain by reducing inflammation. Keywords: chronic low back pain, cytokines, radiofrequency therapy

Toll-Like Receptor 9 Signaling Pathway Contributes to Intestinal Mucosal Barrier Injury in Mice With Severe Acute Pancreatitis.

The purpose of this study was to investigate the role and mechanism of toll-like receptor 9 (TLR9) in intestinal mucosal barrier injury in mice with severe acute pancreatitis (SAP). The mice were randomly divided into 3 groups: control group, SAP group, and TLR9 antagonist-treated group. The expression of tumor necrosis factor-α, interleukin-1β, interleukin-6, diamine oxidase, and endotoxin core antibodies were detected by enzyme-linked immunosorbent assay. The protein expression of zonula occluden-1 (ZO)-1, occludin, TLR9, myeloid differentiation factor 88 (MyD88), tumor necrosis factor receptor-associated factor 6 (TRAF6), p-nuclear factor (NF)-κB p65, and NF-κB p65 were detected by Western blot. TdT-mediated dUTP nick-end labeling staining was used for detecting intestinal epithelial cell apoptosis. The expression of TLR9 and its related pathway proteins MyD88, TRAF6, and p-NF-κB p65 in the intestinal tract of SAP mice were significantly increased compared with that of control mice. Inhibition of the TLR9 expression could reduce the level of serum proinflammatory cytokines, reduce the apoptosis of intestinal epithelial cells, improve intestinal permeability, and ultimately reduce the damage of intestinal mucosal barrier function in SAP. Toll-like receptor 9/MyD88/TRAF6/NF-κB signaling pathway plays an important role in intestinal mucosal barrier injury of SAP.

S1396 Response-Guided Ascites Mobilization in Patients of Acute on Chronic Liver Failure (ACLF) With Acute Kidney Injury Treated With Slow Albumin, Furosemide and Vasoconstrictor Therapy (SAFI+T) versus Standard Treatment

Introduction: Acute-on-chronic liver failure (ACLF) with increasing organ failure is associated with poor outcome. Severely deranged systemic hemodynamic and decreased effective arterial blood volume contributes to tissue damage and organ failure. Response guided therapy with albumin, vasoconstrictors and furosemide may help overcome effective hypovolemia improve diuresis and impact survival. Methods: In observation cohort, 56 patients with ACLF (CANONIC criteria) complicated with AKI ( S.Cr ³ 1.5, >50% increased S.cr from baseline), ascites (≥ Grade II) were enrolled. 30 patients (GROUP I) received response guided(Urine sodium >80mmol/day) slow albumin-furosemide infusion ± terlipressin (SAFI±T), while 26 patients (GROUP II) received standard medical therapy. 28-day survival, ascites mobilization (nil or grade 1), reversal of AKI and adverse events were noted. Laboratory evidences for improvement in various pathophysiological alterations; endotoxemia, cytokine storm, neutrophil dysfunction and secondary infections, following SAFI±T were evaluated. Results: Alcohol and sepsis were the commonest etiologies for chronic and acute insult respectively. Ascites was completely mobilized in 18/30 (60%) patients in Group I (SAFI±T) and 7/26 (26.9%) in Group II (p< 0.05). At 28 days the survival analysis showed, Group I 23/30 (76.4%) and Group II 14/26 (53.8%) (P=< 0.05). There was a lower incidence of hospital acquired infection in Group I of 6.7% vs 38.4% in Group II (p=0.05) and the same was reflected in serum endotoxin and pro-inflammatory cytokine levels; IL-6 and IL-1b showed significant reduction in Group I. Serum endotoxin and TNF-a level increased in Group II despite SMT while it showed significant fall in SAFI+T arm post treatment. Renal artery resistive index was significantly lower after SAFI(T) therapy (0.84 to 0.72) (p< 0.05) as similar trend emulated in UNa level. Conclusion: Patients with ACLF treated with SAFI+T protocol had higher natriuresis, enhanced urine output, complete ascites removal, and improved renal function with a faster mean duration of treatment and better 28-day survival. An in-hospital infection rate was lower. Ascites mobilization was associated with improvement in gut permeability, endotoxemia, pro-inflammatory cytokines, and NETosis in the SAFI+T cohort, showing an association between ascites mobilization and improved pathophysiologic dysfunctions.

Systemic inflammatory response in rats with explosive wound

Aim. To evaluate the temporal changes in circulating cytokines rallying systemic inflammatory response after a blast injury.Material and Methods. Experiments were performed on 60 male Wistar rats (50 rats with blast-related soft tissue injury of the thigh and 10 intact rats). Blast injury was modeled under general anaesthesia using grater firecrackers stuffed with a pyrotechnic mixture. After 3, 7, 14 and 28 days post injury, we carried out a gross examination of the wound and its drainage, measured wound area, assessed local oedema severity, and evaluated serum potassium, myoglobin TNF-α, IL-1β, IL-6, and IL-10.Results. Three to seven days post injury, therewas abundant serous and purulent exudate and pronounced local edema without considerable changes in the wound area. Serum myoglobin and potassium were raised 2.2-2.6-fold and 1.3-fold, respectively (p &lt; 0.05); in addition, we documented the elevation of a major pro-inflammatory cytokine IL-1β. Fourteen days post injury, wound area reduced by 24%, (p &lt; 0.05) along with the decreased wound drainage and local edema of the injured area. Although the levels of serum myoglobin and pro-inflammatory cytokines (in particularIL-6) lowered, they were still significantly higher as compared with the intact rats. Twenty eight days post injury, fibrosis has been completed, epithelialization occurred and only a small scab remained on the wound surface. Yet, serum IL-1β still was 1.3-fold higher than in the intact animals (p &lt; 0.05), albeit IL-6 and TNF-α returned to the reference values. The concentration of anti-inflammatory cytokine IL-10 reached its maximum values relative to the control (3.9-fold, p &lt; 0.05) to the 14th day post injury that was followed by a considerable reduction (though not reaching the reference values) by the period of scar formation.Conclusion. Blast injury in rats is accompanied by a dysregulated balance between pro-inflammatory and anti-inflammatory cytokines throughout all regeneration phases.