Inflammatory Cytokine Profile Research Articles

Immune and inflammatory cytokines profile in Iraqi patients with acute and chronic myeloid leukemia

Introduction and Aim: Evaluation of the profile of immunological and inflammatory cytokines, IL-6, IL-2 and TNF? in acute myeloid leukemia (AML) and chronic myeloid leukemia (CML) patients compared their level with healthy controls, and assessment of the association of these cytokines with clinical and hematological variables. Patients and Methods: Thirty AML, thirty CML patients, and thirty healthy control individuals were recruited for this study from the National Hematology Center for Research and Treatment in Baghdad for the period of October 2020 to March 2021. Leukemic patients were under chemotherapy. Complete blood picture routine test and cytokines profile test were done. IL-6, IL-2 and TNF-? were quantitatively measured using enzymes linked immunosorbent assay (ELISA) kits. Results: Hematological myeloid cancers affect more males than female, in AML male/female ratio was 17/13 while in CML was 16/14. The range of duration of the disease for both AML and CML was 1-3 years. in both AML group the level of IL-6 59.66 ± 31.99 pg/mL and in CML group 57.07± 16.52 pg/mL were both significantly higher P value < 0.001 than that of control group 37.95± 6.10 pg/mL. TNF? in AML group was significantly higher P value < 0.001 than that of control group. A high significant difference in serum level of IL-2 P value <0.0001 in AML and CML compared to controls. Conclusion: IL-6, TNF? and IL-2 levels were higher in myeloid malignancy groups than controls. Serum cytokines shown to be positively associated with each other and could signify to poor outcome of the myeloid malignancy patients.

Inflammatory Cytokine Profiles Do Not Differ Between Patients With Idiopathic Cytopenias of Undetermined Significance and Myelodysplastic Syndromes.

Immune dysregulation has been highlighted as a key player in the pathogenesis of myelodysplastic syndromes (MDS), but little is known about cytokine profiles in patients with unexplained cytopenia with or without mutations in MDS-associated genes (clonal cytopenias of undetermined significance [CCUS] and idiopathic cytopenias of undetermined significance [ICUS], respectively), which often precede MDS. Here, we study the cytokine profiles in 111 patients with ICUS (N = 41), CCUS (N = 30), lower-risk MDS (LR-MDS; N = 22) and higher-risk MDS (HR-MDS; N = 18), and in healthy elderly controls (N = 21). Twenty cytokines were examined in blood plasma at time of diagnosis using Luminex assays and enzyme linked immunosorbent assays. The cytokine levels were compared between patient groups, and in patients versus controls. Associations between cytokines and MDS-associated mutations were evaluated. An aberrant cytokine profile was observed in all patient groups relative to healthy elderly controls. Patients had significantly higher levels of IL-6 (P< 0 .001), tumor necrosis factor α (P < 0.001), IL-10 (P < 0.001), and C-X-C motif chemokine 10 (P < 0.001) and lower levels of transforming growth factor beta 1 (P < 0.001), CCL5/regulated on activation normal T-cell expressed and secreted (P < 0.001), and S100A4 (P < 0.001) compared with healthy controls. Survival was significantly shorter in CCUS and MDS patients with a high systemic inflammatory cytokine load (median overall survival [OS] 21 months) compared with those with low-moderate systemic inflammatory cytokine load (median OS 64 months; P < 0.0001). These data suggest that patients with ICUS and CCUS have cytokine levels as abnormal as in LR-MDS. Indeed, high cytokine levels are present before MDS is diagnosed and cytokine levels are elevated irrespective of the presence or size of the myeloid clones. Cytokines may have a prognostic impact at a very early premalignant stage of myeloid disorders.

Determination of CD91 intracellular signaling pathway following HSP stimulation

Abstract Cancer immunosurveillance is dependent on the heat shock protein (HSP) receptor, CD91, which is expressed on professional antigen-presenting cells (APCs). While CD91-facilitated cross-presentation of HSP-chaperoned peptides has been well studied, CD91-mediated signaling is not completely defined. The binding of CD91 by immunogenic HSPs leads to production of unique inflammatory cytokine profiles by APCs. This requires the phosphorylation of tyrosine residues on intracellular domains of CD91 and activation of NF-κB. However, adaptor proteins associated with CD91 and molecules involved in signal transmission have not been characterized and their role in cancer immunosurveillance remains untested. Our general strategy to identify adaptor proteins involves crosslinking and co-immunoprecipitation of CD91 and its associated signaling complex following stimulation of CD91 with the immunogenic HSPs gp96, calreticulin, and hsp70. Preliminary studies with gp96 have identified Shc as an adaptor protein and AXL receptor tyrosine kinase and tyrosine-protein kinase Fgr as membrane associated kinases. The impact on their inhibition on cytokine release by APCs is being tested. We have also developed a novel mouse expressing mutant, non-signaling CD91, and its impact on cancer immunosurveillance will be determined. Investigation of these complexes will aid in understanding the signaling pathway that activates production of cytokine profiles within APCs. This project will allow us understand the pathways to priming immune responses responsible for cancer immunosurveillance and also to develop better therapeutic strategies for cancer and autoimmune diseases. Supported by grants from NIH (RO1 CA233803)

Inflammatory cytokines analysis in C57BL/6 mouse skin after dermal exposure with chemical threat agent phosgene oxime

Phosgene Oxime (dichloroform oxime; CX), an urticant categorized as a vesicating agent, is a potential chemical threat agent. Its exposure causes rapid and painful dermal injury and systemic toxic effects. CX exposure can result in enzyme inactivation, corrosive injury, and cell death with rapid tissue destruction, and induce the recruitment of immune cells. However, the mechanism of CX‐induced skin toxicity and inflammation is not known. Studies in our lab have shown that CX exposure causes mast cell degranulation and release of inflammatory mediators like COX‐2, MMP‐9, myeloperoxidase, and an inflammatory response in the skin. In the present study, we analyzed the inflammatory cytokine profile in male and female C57BL/6 mouse skin following dermal CX exposure (neat CX for either 0.5 or 1.0 min using two 12 mm vapor caps on the dorsal skin at MRIGlobal). Cytokine array analyses showed an increase in the pro‐inflammatory cytokines IL‐6 (Interleukin 6) and TNF‐α (Tumor Necrosis Factor Alpha) in both male and female mice at 24h post CX‐exposure. KC (neutrophil chemoattractant), MCP‐1 (Monocyte chemoattractant), MCSF (Macrophage colony‐stimulating factor) and RANTES (T cell, monocyte attractant) increased in both sexes at day 3 post CX‐exposure, showing involvement of immune cells in the CX‐induced injury. IL‐13, an allergic inflammation and several diseases’ mediator was found to be increased at later time point in female mice and at 2 hours after 1.0 min CX exposure in male mice. IL‐4 and IL‐5 levels decreased at 24 h but were increased at day 3 post 0.5 min CX exposure. Altogether, our results show immune cell infiltration with changes in inflammatory cytokines. Similar Inflammatory cytokine changes from CX exposure were observed in male and female mice; however, slightly different patterns were evident in some cytokines in female mice that related to the observed difference in the inflammatory response and somewhat faster dermal wound healing. Further molecular analysis is underway to determine the signaling pathways involved in CX‐induced dermal inflammation and toxicity for identifying the therapeutic targets.

Differences of inflammatory cytokine profile in patients with vulnerable plaque: A coronary CTA study

Background and aimsVarious pro- and anti-inflammatory biomarkers are involved in the process of atherosclerosis. We analyzed the association of different biomarkers with coronary plaque volume and vulnerable plaque subcomponents. MethodsIn 301 patients undergoing coronary CT angiography (CTA), total coronary plaque volume (TPV) and subcomponents including non-calcified plaque volume (NCPV) and vulnerable plaque burden were quantified using semi-automated software. Serum was analyzed for various cytokines. ResultsOut of 301 patients, 207 (69%) were male. The mean age was 59 ± 10 years. Patients were divided using the median of TPV, NCPV and vulnerable plaque burden. In univariable analysis, patients with high TPV, high NCPV and high vulnerable plaque burden showed significant higher serum levels for IFNƔ, IL-1a, −2, −4, −10 and −17 and significant lower levels for IL-8 and MCP-1 (all p < 0.05). Multivariable analysis showed positive associations between high vulnerable plaque burden, IL-1a (OR 2.60, p = 0.001) and Eotaxin (OR 1.89, p = 0.020), and inverse association to MCP-1 (OR 0.33, p < 0.001), independent of age, gender and CVRF.In exploratory subanalyses, patients with presence of atherosclerosis (n = 247; 82%) showed significantly higher levels of IL-17 in all subgroups with high vulnerable plaque burden, irrespective of overall plaque volume (all p < 0.001). ConclusionsThe cytokine profile significantly differs between patients with high and low coronary plaque volume. IL-1a and IL-17 seem to play a major proatherogenic role in vulnerable plaque formation, whereas MCP-1 paradoxically portends protective effects. Longitudinal studies with serial cytokine testing are needed to identify potential targets for therapeutic interventions.

CORRELATION BETWEEN INFLAMATORY CYTOKINES AND ANTHROPOLOGICAL PARAMETERS IN THE AGED POPULATION

Cytokines play an important role in regulating the immune response in the elderly population, and various studies indicate an inability to control systemic inflammation in the elderly, which may be a sign of ‘unsuccessful aging’. Thus, the evaluation of inflammatory cytokine profile in a healthy elderly population is a prerequisite for assessing the risk of developing the disease. The aim of the present study was to evaluate the level of cytokines - IL-6 and IL-18 in the healthy elderly population and to reveal their correlation with anthropological parameters. The results of our study shown that IL-18 levels were statistically significantly correlated with height in men and systolic blood pressure in women. IL-18 levels in the general population are reliably and strongly associated with IL-6 (r = 0.475; p = 0.001). Thus, based on our findings, evaluation of IL-6 and IL-18 concentrations can be used as biomarkers of “inflamaging”.

Proteomic analysis of the periodontal pathogen Prevotella intermedia secretomes in biofilm and planktonic lifestyles

Prevotella intermedia is an important species associated with periodontitis. Despite the remarkable clinical significance, little is known about the molecular basis for its virulence. The aim of this study was to characterize the secretome of P. intermedia in biofilm and planktonic life mode. The biofilm secretome showed 109 proteins while the planktonic secretome showed 136 proteins. The biofilm and the planktonic secretomes contained 17 and 33 signal-peptide bearing proteins, 13 and 18 lipoproteins, respectively. Superoxide reductase, sensor histidine kinase, C40 family peptidase, elongation factor Tu, threonine synthase etc. were unique to biofilm. Of the ~ 30 proteins with predicted virulence potential from biofilm and planktonic secretomes, only 6 were common between the two groups, implying large differences between biofilm and planktonic modes of P. intermedia. From Gene Ontology biofilm secretome displayed a markedly higher percent proteins compared to planktonic secretome in terms of cellular amino acid metabolic process, nitrogen compound metabolic process etc. Inflammatory cytokine profile analysis revealed that only the biofilm secretome, not the planktonic one, induced important cytokines such as MIP-1α/MIP-1β, IL-1β, and IL-8. In conclusion, the revealed differences in the protein profiles of P. intermedia biofilm and planktonic secretomes may trigger further questions about molecular mechanisms how this species exerts its virulence potential in the oral cavity.

Impact of Vitamin C on Gene Expression Profile of Inflammatory and Anti-Inflammatory Cytokines in the Male Partners of Couples with Recurrent Pregnancy Loss

Immune system disorders and increased inflammation in the male reproductive system can lead to fetal risk in the early stages of development and implantation. Antioxidants such as vitamin C can play a protective role against sperm inflammatory reactions. This study aimed to evaluate the effect of vitamin C on the expression of inflammatory and anti-inflammatory cytokine genes in the male partners of couples with recurrent pregnancy loss. In this randomized clinical trial, twenty male partners of couples with RPL were examined for sperm parameters and expression profile of some inflammatory and anti-inflammatory cytokine genes before and after treatment with vitamin C. There was a statistically significant higher rate of normal morphology and sperm concentration in each patient before and after treatment with vitamin C (p ≤ 0.05). The mRNA levels of interleukin 6 and tumor necrosis factor-alpha were significantly decreased in the sperm of patients after treatment with vitamin C compared to before treatment. In contrast, the gene expression levels of interleukin 4 and transforming growth factor-beta showed a significant increase in the sperm of patients after treatment with vitamin C. Oral daily administration of vitamin C may be effective in the fertility potential of male partners of couples with RPL not only through the improvement of the sperm parameters but also by modulating the expression profile of inflammatory and anti-inflammatory genes. Further studies on protein levels are needed to clarify the role of TNF-⍺ and IFN-γ as a prognostic value in evaluating the recurrent abortion risk in infertile male partners. This trial is registered with IRCT20180312039059N1.

Age-Dependent Reduction in Asthmatic Pathology through Reprogramming of Postviral Inflammatory Responses.

Asthma is a chronic disease of childhood, but for unknown reasons, disease activity sometimes subsides as children mature. In this study, we present clinical and animal model evidence suggesting that the age dependency of childhood asthma stems from an evolving host response to respiratory viral infection. Using clinical data, we show that societal suppression of respiratory virus transmission during coronavirus disease 2019 lockdown disrupted the traditional age gradient in pediatric asthma exacerbations, connecting the phenomenon of asthma remission to virus exposure. In mice, we show that asthmatic lung pathology triggered by Sendai virus (SeV) or influenza A virus is highly age-sensitive: robust in juvenile mice (4-6 wk old) but attenuated in mature mice (>3 mo old). Interestingly, allergen induction of the same asthmatic traits was less dependent on chronological age than viruses. Age-specific responses to SeV included a juvenile bias toward type 2 airway inflammation that emerged early in infection, whereas mature mice exhibited a more restricted bronchiolar distribution of infection that produced a distinct type 2 low inflammatory cytokine profile. In the basal state, aging produced changes to lung leukocyte burden, including the number and transcriptional landscape of alveolar macrophages (AMs). Importantly, depleting AMs in mature mice restored post-SeV pathology to juvenile levels. Thus, aging influences chronic outcomes of respiratory viral infection through regulation of the AM compartment and type 2 inflammatory responses to viruses. Our data provide insight into how asthma remission might develop in children.

Liver transplantation for Hepatocellular Carcinoma: A prognostic model incorporating pretransplant inflammatory cytokines

PurposeLiver transplantation (LT) remains an optimal treatment for selected hepatocellular carcinoma (HCC). Cytokines can be obtained by minimally invasive techniques and are associated with the development of HCC. The purpose of our investigation was to explore the prognostic value of pretransplant serum inflammatory cytokine profiles in HCC patients for LT. MethodsWe detected forty inflammatory cytokines in pre-LT serum from 42 HCC patients by using an inflammation-related antibody array. A pretransplant serum inflammatory cytokine-associated risk assessment model (pre-SCRAM) was developed and was validated in an external cohort of 213 HCC patients who underwent LT and were then followed prospectively. ResultsThe pre-LT factors independently associated with recurrence-free survival (RFS) were as follows: B-lymphocyte chemoattractant (BLC), interleukin (IL)-12p40 and maximum tumor diameter. High IL-12P40 level was associated with a significantly smaller maximum tumor diameter (p = 0.021), decreased proportion of nodules ≥ 3 (p = 0.001), lower platelet counts (p = 0.011) and lower portal vein tumor thrombus (p = 0.031). Conversely, recipients with poor BLC level had higher alpha-fetoprotein (AFP) levels (p = 0.016). Kaplan–Meier analyses revealed that high pre-LT BLC or IL-12p40 level was associated with superior RFS. The pre-SCRAM stratified recipients into three risk groups: high risk, intermediate risk and low risk. In the validation cohort, for patients in the high, intermediate, and low risk groups, the 3-year RFS rates were 29.3%, 58.7%, and 82.2%, respectively, the 3-year HCC-specific survival rates were 54.5%, 73.8%, and 86%, respectively, and the 3-year overall survival rates were 44.4%, 60.9%, and 79.9%, respectively. The pre-SCRAM model performed well and remained significant in optimizing the risk stratification of recurrence in patients beyond the Milan criteria or the AFP model. ConclusionPretransplant cytokine profiles can provide powerful prognostic information in the setting of LT for HCC. A pre-LT risk model incorporating cytokines showed excellent efficiency in recurrence prediction for HCC patients, which could ultimately stratify the prognosis in patients beyond the Milan criteria or the AFP model.

Investigating the inflammatory status in the patients candidate for second angiography after coronary stent implantation

Background Inflammation has been shown to be an important feature of atherosclerosis. We aimed to assess a profile of inflammatory cytokines and growth factors in patients with established coronary artery disease (CAD), 12 months after stent implantation. Methods A total of 193 patients with CAD, who were candidates for angiography, 12 months after stent implantation (cases), were compared with 107 patients with CAD, who were candidates for their first angiography (controls). Fasting blood glucose (FBG), triglycerides (TGs), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and high-sensitive C-reactive protein (hs-CRP) were measured using routine methods. The serum concentrations of IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, TNF-α, IFN-γ, MCP-1, EGF and VEGF were determined using competitive chemiluminescence immunoassays. Results Serum levels of FBG (p = .002), TG (p = .029) and hs-CRP (p = .005) were significantly lower in cases than controls. The cytokines and growth factor profiles in cases were significantly different from controls. After multivariate analysis, serum levels of IL-2 (p < .001), IL-4 (p = .028) were significantly lower in cases compared with the controls while serum levels of IL-8, TNF-α, MCP-1, EGF and VEGF were significantly higher in the cases (p < .001). Conclusions In patients with CAD and higher consumption of drug used (statins, aspirin and glucose lowering agents) to mitigate the risk of a secondary event, the level of hs-CRP one year after stent implantation decreased despite of significant higher serum levels of pro- and anti-inflammatory cytokines and growth factors.

The expression analysis of IL-6, IL-18, IL-21, IL-23, and TGF-β mRNA in the nasal mucosa of patients with Allergic rhinitis.

The profile of inflammatory and suppressing cytokines is important to contribute to the disruption of TH1/TH2 balance in Allergic rhinitis (AR). This study aimed to assess the expression levels of IL-6, IL-18, IL-21, IL-23, and TGF-β in nasal biopsies in AR patients and evaluate its correlation with the severity of AR. The study included 30 patients with mild persistent allergic rhinitis (MPAR), patients with moderate-to-severe (M/S) PAR, and 30 healthy individuals. The biopsies of nasal inferior turbinate mucosa were collected from each participant. The expression of IL-6, IL-18, IL-21, IL-23, and TGF-β was evaluated by the quantitative real-time polymerase chain reaction. The degree of eosinophil infiltration into the nasal mucosa, blood eosinophils, and total serum IgE level were also measured. The expression of IL-6, IL-18, and IL-23 in patients with AR significantly increased compared to the control group. Conversely, the gene expression of the TGF-β declined in the M/S PAR group rather than the AR- group. The data did not show a significant difference in the expression of the IL-21 gene between AR+ and AR- groups. We suggested that inflammatory cytokines including IL-6, IL-18, and IL-23 may be involved in the severity of AR and associated with markers of inflammation.

Abstract 151: Association of vegetarian and non-vegetarian diet with inflammatory biomarkers in healthy North Indian population

Background: Low-grade chronic inflammation is a known risk factor for the development of age-related diseases such as CVD, diabetes and some cancers, these findings have stimulated research into the influence of nutrition and dietary patterns on inflammatory indices.Aims and Objective: To assess the effect of plant based vs. animal origin diets on serum markers of inflammation in normal healthy individuals.Results: 275 non-vegetarians and 353 vegetarians were included in the study. Weight and BMI was significantly higher in the vegetarians. Both fasting blood glucose and Insulin level were significantly higher in vegetarians, while total cholesterol, HDL, triglyceride and LDL was significantly higher in the non-vegetarians. Serum ALT and ALP was significantly higher and total protein and albumin was lower in vegetarian group. The serum level of IL-6, IL-1, resistin and CRP was significantly higher in vegetarian, while the levels of IL-10, adiponectin were significantly raised in non-vegetarian group. BMI had significant positive correlation with TNFα, IL-6, Resistin, hsCRP and IL-1 β, protein intake had significant negative correlation with TNFα, IL-6, and hsCRP.Conclusion: Vegetarian diet in Indians is associated with higher BMI, blood glucose and insulin level. Non-vegetarian diet is associated with a more favorable inflammatory cytokine profile.

Diagnosis of sepsis with inflammatory biomarkers, cytokines, endothelial functional markers from SIRS patients.

BackgroundSepsis is a life-threatening illness with a challenging diagnosis. Rapid detection is the key to successful treatment of sepsis. To investigate diagnostic value, the plasma protein profiles of inflammatory biomarkers, cytokines, and endothelial functional markers were compared between healthy controls, SIRS, and septic patients.MethodsThe plasma protein profiles were performed by Luminex Assay in a cohort of 50 SIRS patients, 82 septic patients and 25 healthy controls. Fourteen plasma proteins were analyzed in the same cohort: IL-1β, IL-6, IL-8, IL-10, CCL-2, VEGF, VEGF-C, VEGFR2, CD62E, CD62P, MFG-E8, ICAM-1, TFPI, Urokinase.ResultIL-2R, IL-6, IL-8, IL-10, CCL-2, ICAM-1, and Urokinase were significantly higher in sepsis patients than SIRS patients. VEGF, IL-1β, CD62E, CD62P, MFG-E8, and TFPI have no statistical difference. VEGF-C, VEGFR2 were significantly different in SIRS patients than sepsis patients. Urokinase, ICAM-1, and VEGFR2 were significantly different between sepsis group and SIRS group. The AUCs of Urokinase, ICAM-1, and VEGFR2 and the combination for the diagnosis of sepsis were 0.650, 0.688, 0.643, and 0.741, respectively.ConclusionsMost patients have the higher level of several cytokines and developed endothelial cell injury in the initial phase of sepsis, Urokinase, ICAM-1, and VEGFR2 may be useful to evaluate severity and prognosis of sepsis patients.

IFIT2-depleted metastatic oral squamous cell carcinoma cells induce muscle atrophy and cancer cachexia in mice.

BackgroundInterferon‐induced protein with tetratricopeptide repeat 2 (IFIT2) is a reported metastasis suppressor in oral squamous cell carcinoma (OSCC). Metastases and cachexia may coexist. The effect of cancer metastasis on cancer cachexia is largely unknown. We aimed to address this gap in knowledge by characterizing the cachectic phenotype of an IFIT2‐depleted metastatic OSCC mouse model.MethodsGenetically engineered and xenograft tumour models were used to explore the effect of IFIT2‐depleted metastatic OSCC on cancer cachexia. Muscle and organ weight changes, tumour burden, inflammatory cytokine profiles, body composition, food intake, serum albumin and C‐reactive protein (CRP) levels, and survival were assessed. The activation of the IL6/p38 pathway in atrophied muscle was measured.ResultsIFIT2‐depleted metastatic tumours caused marked body weight loss (−18.2% vs. initial body weight, P < 0.001) and a poor survival rate (P < 0.01). Skeletal muscles were markedly smaller in IFIT2‐depleted metastatic tumour‐bearing mice (quadriceps: −28.7%, gastrocnemius: −29.4%, and tibialis: −24.3%, all P < 0.001). Tumour‐derived circulating granulocyte‐macrophage colony‐stimulating factor (+772.2‐fold, P < 0.05), GROα (+1283.7‐fold, P < 0.05), IL6 (+245.8‐fold, P < 0.001), IL8 (+616.9‐fold, P < 0.001), IL18 (+24‐fold, P < 0.05), IP10 (+18.8‐fold, P < 0.001), CCL2 (+439.2‐fold, P < 0.001), CCL22 (+9.1‐fold, P < 0.01) and tumour necrosis factor α (+196.8‐fold, P < 0.05) were elevated in IFIT2‐depleted metastatic tumour‐bearing mice. Murine granulocyte colony‐stimulating factor (+61.4‐fold, P < 0.001) and IL6 (+110.9‐fold, P < 0.01) levels were significantly increased in IFIT2‐depleted metastatic tumour‐bearing mice. Serum CRP level (+82.1%, P < 0.05) was significantly increased in cachectic shIFIT2 mice. Serum albumin level (−26.7%, P < 0.01) was significantly decreased in cachectic shIFIT2 mice. An assessment of body composition revealed decreased fat (−81%, P < 0.001) and lean tissue (−21.7%, P < 0.01), which was consistent with the reduced food intake (−19.3%, P < 0.05). Muscle loss was accompanied by a smaller muscle cross‐sectional area (−23.3%, P < 0.05). Muscle atrophy of cachectic IFIT2‐depleted metastatic tumour‐bearing mice (i.v.‐shIFIT2 group) was associated with elevated IL6 (+2.7‐fold, P < 0.05), phospho‐p38 (+2.8‐fold, P < 0.05), and atrogin‐1 levels (+2.3‐fold, P < 0.05) in the skeletal muscle. Neutralization of IL6 rescued shIFIT2 conditioned medium‐induced myotube atrophy (+24.6%, P < 0.01).ConclusionsOur results suggest that the development of shIFIT2 metastatic OSCC lesions promotes IL6 production and is accompanied by the loss of fat and lean tissue, anorexia, and muscle atrophy. This model is appropriate for the study of OSCC cachexia, especially in linking metastasis with cachexia.

An Objective Diagnosis Model with Integrated Metabolic and Immunity Parameters for Phlegm-Dampness Constitution.

Background According to Chinese constitutional theory, people can be divided into nine constitutions, which represent distinctive vulnerability to different diseases such as metabolic syndrome, atherosclerosis, and immunity-related disease, and so forth in modern medicine, phlegm-dampness constitution (PDC) is one of the nine constitutions, which is susceptible to metabolic syndrome (MS) and atherosclerosis that associate with lipid metabolism and immunity dysregulation closely. Objectives In this study, we aimed to investigate the metabolic and immunity profiles of phlegm-damp constitution (PDC), including metabolites, lymphocytes distribution, and inflammatory cytokines. Methods A total of 74 patients with PDC and 66 individuals with gentle constitution (GC) were enrolled in this study. We utilized biochemical methods to detect metabolic parameters, flow cytometry to survey T/B/NK/NKT lymphocyte subgroups distribution, and ELISA to assay inflammatory cytokines. Results The subjects with PDC had higher GLU, AI TC, TG, and LDL-C and lower HDL-C levels. The immunity profile indicated that PDC subjects had higher percentage of WBCs, neutrophils, lymphocytes, B cells, and natural killer T cells compared with subjects with GC (P < 0.05). Serum levels of IL-10 decreased significantly in the subjects with phlegm-damp constitution, whereas IL-12 levels increased dramatically in the PDC group compared with the GC group (both P < 0.05). Additionally, logistic regression identified four independent variables (GLU, TG, LDL-C, and lymphocytes) that were highly correlated with PDC (P < 0.05). The area under the curve of the receiver operating characteristic curve was 0.878, which indicated the data were reliable to distinguish the subjects with PDC from the ones with GC. Conclusion Phlegm-damp constitution was prone to hyperglycemia and hyperlipidemia syndrome, promoting the occurrence and progression of metabolic-related diseases. Interestingly, proinflammatory cells and cytokines were activated in the PDC group as well. Our findings could offer a profile of early screening indicators to identify high-risk patients of metabolic- and immunity-related diseases from Chinese constitution.

2-Benzyllawsone protects against polymicrobial sepsis and vascular hyporeactivity in swiss albino mice

BackgroundNovel naphthoquinone, 2-benzyllawsone (LT-9) was evaluated against vascular hyporeactivity and sepsis in cecal ligation and puncture (CLP) model in mice in view of its preliminary antibacterial and anti-inflammatory properties and to explore whether pretreatment with the molecule could restore vascular tone and contractile response to norepinephrine. MethodsEvaluation of LT-9 against vascular hyporeactivity, hypotension, and sepsis-related inflammation and infection was carried out in the CLP model in Swiss albino mice and aortic smooth muscle cells in vitro. ResultsLT-9 showed potent reversal of the vascular hyporeactivity in CLP mice aorta. The increased contraction response to norepinephrine in CLP mouse aorta by LT-9 was mediated by opening of L-type voltage-dependent calcium channels (VDCC) verified by ex vivo experiment where LT-9 enhanced contraction response to CaCl2 in the aorta while abolishing the contraction response of known VDCC opener Bay K8644. LT-9 in aortic smooth muscle cells showed Fluo-4 mediated increase in calcium fluorescence. Oral administration of LT-9 at 50 and 100 mg kg−1 day−1 for 15 days significantly enhanced the mean survival time, improved hemodynamic and Electrocardiogram (ECG) profile, and aortic tissue reactivity in CLP mice. Further, LT-9 significantly reversed the perturbation of the expression profile of inflammatory cytokines, reduced the splenic microbial load, and was well tolerated in oral toxicity. ConclusionsLT-9 showed potent biological activity against sepsis and was found to be well tolerated in the toxicity study in Swiss albino mice and showed promise for the benzyllawsone class of molecules against sepsis for the development of novel pharmacophore.

Qiguiyin Decoction Improves Multidrug-Resistant Pseudomonas aeruginosa Infection in Rats by Regulating Inflammatory Cytokines and the TLR4/MyD88/NF-κB Signaling Pathway.

Pseudomonas aeruginosa (PA), a Gram-negative bacterium, has a high detection rate in hospital-acquired infections. Recently, the frequent appearance of multidrug-resistant (MDR) PA strain with high morbidity and mortality rates has aggravated the difficulty in treating infectious diseases. Due to its multiple resistance mechanisms, the commonly used antibiotics have gradually become less effective. Qiguiyin decoction (QGYD) is a clinically experienced prescription of Chinese herbal medicine, and its combined application with antibiotics has been confirmed to be effective in the clinical treatment of MDR PA infection, which could be a promising strategy for the treatment of drug-resistant bacterial infections. However, the mechanism of QGYD restoring antibiotics susceptibility to MDR PA remains unclear. In the present study, we investigated the effects of QGYD and levofloxacin (LEV) singly or in combination on MDR PA-induced pneumonia rat models. Further analysis was carried out in the serum differential expression profiles of inflammatory cytokines by cytokine antibody array. Besides, the lung TLR4/MyD88/NF-κB signaling pathway was detected by RT-qPCR. Our results showed that based on the treatment of MDR PA-infected rat model with LEV, the combination of QGYD improved the general state and immune organ index. Furthermore, it moderately increased the expressions of proinflammatory cytokines including IL-1β, IL-6, and TNF-α in the early stage of infection and decreased their release rapidly in the later stage, while regulated the same phase change of anti-inflammatory cytokine IL-10. In addition, the adhesion molecule ICAM-1 was significantly downregulated after QGYD combined with LEV treatment. Moreover, the mRNA expressions of TLR4, MyD88, NF-κB, and ICAM-1 were significantly downregulated. These results indicated that the mechanism of QGYD restoring LEV susceptibility to MDR PA was related to its regulation of inflammatory cytokines and the TLR4/MyD88/NF-κB signaling pathway, which provides theoretical support for the clinical application of QGYD combined with LEV therapy to MDR PA infection.

Influence of adjunctive non-surgical peri‑implant therapy on clinical and salivary cytokine profile in obese patients

AimThe present study aimed to evaluate the impact of Photodynamic therapy (PDT) and Er,Cr:YSGG laser (ECL) adjunct to nonsurgical mechanical debridement (NSMD) in comparison to conventional NSMD in overweight individuals with peri‑implant disease on peri‑implant inflammatory parameters and cytokine levels of IL-6 and TNF-α in Peri-implant crevicular fluid (PICF) Materials and MethodsRecruitment of volunteers was performed from the outpatient department according to predefined inclusion and exclusion criteria. Group 1 consisted of obese participants who underwent peri‑implant mechanical debridement (MD); group 2, obese participants who experienced MD adjunct to ECL, and in group 3, obese participants were treated with MD and adjunct PDT. Clinical periodontal parameters, plaque index (PI), bleeding index (BI) and probing depth (PD) along with crestal bone loss (CBL) were assessed at baseline, 3 months, and 6 months. PICF was collected to evaluate the levels of IL-6 and TNF-α. Kruskal–Wallis and Bonferroni post hoc test were employed for the comparison of peri‑implant inflammatory parameters and PICF cytokine profiles among the study groups. ResultsClinical peri‑implant parameters (PI, BI and PD) at baseline among subjects in group 1, group 2 and group 3 were comparable (p > 0.05). At 3 months follow up, mean scores of peri‑implant PI, BI, and PD (P <0.05) in group 1 (control) were significantly higher compared to group 2 (ECL) and 3 (PDT) (p < 0.05). At 6 months follow-up, PI and BI mean scores among groups 1, 2, and 3 were comparable (p>0.05). At 6 months participants in groups 2 and 3 noted significant difference (p<0.05) compared to baseline with no difference noted in the control group (p>0.05). ConclusionPhotodynamic therapy and Er, Cr: YSGG adjunct to NSMD demonstrated significant improvement in peri‑implant inflammatory parameters in obese individuals. Multicentric clinical trials are suggested to extrapolate the findings of the present study.

Identification of SARS-CoV-2 Variants and Their Clinical Significance in Hefei, China.

The ongoing coronavirus disease 2019 (COVID-19) pandemic represents one of the most exigent threats of our lifetime to global public health and economy. As part of the pandemic, from January 10 to March 10, 2020, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) began to spread in Hefei (Anhui Province, China) with a total of 174 confirmed cases of COVID-19. During this period, we were able to gather critical information on the transmission and evolution of pathogens through genomic surveillance. Particularly, the objective of our study was to track putative variants of SARS-CoV-2 circulating in Hefei for the first time and contribute to the global effort toward elucidating the molecular epidemic profile of the virus. Patients who showed symptoms of COVID-19 were routinely tested for SARS-CoV-2 infections via RT-PCR at the First Affiliated Hospital of Anhui Medical University. Whole-genome sequencing was performed on 97 clinical samples collected from 29 confirmed COVID-19 patients. As a result, we identified a local novel single-nucleotide polymorphism site (10,380) harboring a G → T mutation (Gly → Val) in Hefei. Further phylogenetic network analysis with all the sequences of SARS-CoV-2 deposited in GenBank collected in East and Southeast Asia revealed a local subtype of S-type SARS-CoV-2 (a1) harboring a C → T synonymous mutation (Leu) at position 18,060 of ORF1b, likely representing a local SARS-CoV-2 mutation site that is obviously concentrated in Hefei and the Yangtze River Delta region. Moreover, clinical investigation on the inflammatory cytokine profile of the patients suggested that mutations at positions 18,060 (the shared variable site of subtype a1) and 28,253(harboring a C → T synonymous mutation, Phe) were associated with milder immune responses in the patients.