Single-cell transcriptome profiling of sepsis identifies HLA-DR^(low)S100A^(high)monocytes with immunosuppressive function

Background Sustained yet intractable immunosuppression is commonly observed in septic patients,resulting in aggravated clinical outcomes.However,due to the substantial heterogeneity within septic patients,precise indicators in deciphering clinical trajectories and immunological alterations for septic patients remain largely lacking.Methods We adopted cross-species,single-cell RNA sequencing(scRNA-seq)analysis based on two published datasets containing circulating immune cell profile of septic patients as well as immune cell atlas of murine model of sepsis.Flow cytometry,laser scanning confocal microscopy(LSCM)imaging and Western blotting were applied to identify the presence of S100A9^(+)monocytes at protein level.To interrogate the immunosuppressive function of this subset,splenic monocytes isolated from septic wild-type or S100a9^(–/–)mice were co-cultured with naive CD4^(+)T cells,followed by proliferative assay.Pharmacological inhibition of S100A9 was implemented using Paquinimod via oral gavage.Results scRNA-seq analysis of human sepsis revealed substantial heterogeneity in monocyte compartments following the onset of sepsis,for which distinct monocyte subsets were enriched in disparate subclusters of septic patients.We identified a unique monocyte subset characterized by high expression of S100A family genes and low expression of human leukocyte antigen DR(HLA-DR),which were prominently enriched in septic patients and might exert immunosuppressive function.By combining single-cell transcriptomics of murine model of sepsis with in vivo experiments,we uncovered a similar subtype of monocyte significantly associated with late sepsis and immunocompromised status of septic mice,corresponding to HLA-DR^(low)S100A^(high)monocytes in human sepsis.Moreover,we found that S100A9^(+)monocytes exhibited profound immunosuppressive function on CD4^(+)T cell immune response and blockade of S100A9 using Paquinimod could partially reverse sepsis-induced immunosuppression.Conclusions This study identifies HLA-DR^(low)S100A^(high)monocytes correlated with immunosuppressive state upon septic challenge,inhibition of which can markedly mitigate sepsis-induced immune depression,thereby providing a novel therapeutic strategy for the management of sepsis.

The role of monocyte-lineage cells in human immunodeficiency virus persistence: mechanisms and progress

Human immunodeficiency virus type 1(HIV-1) persistence is a major barrier to the successful treatment and eradication of acquired immunodeficiency syndrome(AIDS).In addition to resting CD4+ T cells,a significant long-lived compartment of HIV-1 infection in vivo includes blood monocytes and tissue macrophages.Studying HIV-1 persistence in monocyte-lineage cells is critical because these cells are important HIV-1 target cells in vivo.Monocyte-lineage cells,including monocytes,dendritic cells(DCs) and macrophages,play a significant role in HIV-1 infection and transmission.These cells have been implicated as viral reservoirs that facilitate HIV-1 latency and persistence.A better understanding of HIV-1 interactions with monocyte-lineage cells can potentially aid in the development of new approaches for intervention.This minireview highlights the latest advances in understanding the role of monocyte-lineage cells in HIV-1 persistence and emphasizes new insights into the mechanisms underlying viral persistence.

Malarial pigment does not induce MMP-2 and TIMP-2 protein release by human monocytes

Dear editor, In the recent years growing evidence on the involvement of human matrix metalloproteinases(MMPs) and tissue inhibitors of metalloproteinases(TIMPs) in cerebral malaria (CM) has been reported[1]and a role for malarial pigment haemozoin(HZ) has been proposed[2,3].In a recent work my group showed that in human microvascular endothelial

Inhibition of matrix metalloproteinase-9 secretion by dimethyl sulfoxide and cyclic adenosine monophosphate in human monocytes

BACKGROUND Matrix metalloproteinases(MMPs),including MMP-9,are an integral part of the immune response and are upregulated in response to a variety of stimuli.New details continue to emerge concerning the mechanistic and regulatory pathways that mediate MMP-9 secretion.There is significant evidence for regulation of inflammation by dimethyl sulfoxide(DMSO)and 3',5'-cyclic adenosine monophosphate(cAMP),thus investigation of how these two molecules may regulate both MMP-9 and tumor necrosis factorα(TNFα)secretion by human monocytes was of high interest.The hypothesis tested in this study was that DMSO and cAMP regulate MMP-9 and TNFαsecretion by distinct mechanisms.AIM To investigate the regulation of lipopolysaccharide(LPS)-stimulated MMP-9 and tumor necrosis factorαsecretion in THP-1 human monocytes by dimethyl sulfoxide and cAMP.METHODS The paper describes a basic research study using THP-1 human monocyte cells.All experiments were conducted at the University of Missouri-St.Louis in the Department of Chemistry and Biochemistry.Human monocyte cells were grown,cultured,and prepared for experiments in the University of Missouri-St.Louis Cell Culture Facility as per accepted guidelines.Cells were treated with LPS for selected exposure times and the conditioned medium was collected for analysis of MMP-9 and TNFαproduction.Inhibitors including DMSO,cAMP regulators,and anti-TNFαantibody were added to the cells prior to LPS treatment.MMP-9 secretion was analyzed by gel electrophoresis/western blot and quantitated by ImageJ software.TNFαsecretion was analyzed by enzyme-linked immuno sorbent assay.All data is presented as the average and standard error for at least 3 trials.Statistical analysis was done using a two-tailed paired Student t-test.P values less than 0.05 were considered significant and designated as such in the Figures.LPS and cAMP regulators were from Sigma-Aldrich,MMP-9 standard and antibody and TNFαantibodies were from R&D Systems,and amyloid-βpeptide was from rPeptide.RESULTS In our investigation of MMP-9 secretion from THP-1 human monocytes,we made the following findings.Inclusion of DMSO in the cell treatment inhibited LPSinduced MMP-9,but not TNFα,secretion.Inclusion of DMSO in the cell treatment at different concentrations inhibited LPS-induced MMP-9 secretion in a dosedependent fashion.A cell-permeable cAMP analog,dibutyryl cAMP,inhibited both LPS-induced MMP-9 and TNFαsecretion.Pretreatment of the cells with the adenylyl cyclase activator forskolin inhibited LPS-induced MMP-9 and TNFαsecretion.Pretreatment of the cells with the general cAMP phosphodiesterase inhibitor IBMX reduced LPS-induced MMP-9 and TNFαin a dose-dependent fashion.Pre-treatment of monocytes with an anti-TNFαantibody blocked LPSinduced MMP-9 and TNFαsecretion.Amyloid-βpeptide induced MMP-9 secretion,which occurred much later than TNFαsecretion.The latter two findings strongly suggested an upstream role for TNFαin mediating LPS-stimulate MMP-9 secretion.CONCLUSION The cumulative data indicated that MMP-9 secretion was a distinct process from TNFαsecretion and occurred downstream.First,DMSO inhibited MMP-9,but not TNFα,suggesting that the MMP-9 secretion process was selectively altered.Second,cAMP inhibited both MMP-9 and TNFαwith a similar potency,but at different monocyte cell exposure time points.The pattern of cAMP inhibition for these two molecules suggested that MMP-9 secretion lies downstream of TNFαand that TNFαmay a key component of the pathway leading to MMP-9 secretion.This temporal relationship fit a model whereby early TNFαsecretion directly led to later MMP-9 secretion.Lastly,antibody-blocking of TNFαdiminished MMP-9 secretion,suggesting a direct link between TNFαsecretion and MMP-9 secretion.

Decreased human leukocyte antigen-D-related expression on CD14^(+)monocytes in patients with out-of-hospital cardiac arrest provided target temperature management therapy:a prospective observational study

Background:Post–cardiac arrest syndrome involves systemic inflammation,which causes subsequent neurological impairments.We investigated the influence of targeted temperature management(TTM)therapy in patients with out-of-hospital cardiac arrest(OHCA)after return of spontaneous circulation(ROSC)by observing the changes in circulating CD14^(+)monocytes and the expression of human leukocyte antigen D–related(HLA-DR)and programmed cell death ligand 1(PD-L1)in CD14^(+)monocytes.Methods:Adult patients admitted to the emergency department of Beijing Chao-Yang Hospital after OHCA between January 2017 and March 2018 were included in this study.Thirty control subjects,10 patients with OHCA,and 37 patients with OHCA who received 72 hours of TTM therapy were enrolled.Peripheral blood samples of patients in the OHCA and TTM groups were collected on Days 1 and 3(D1 and D3)after ROSC and evaluated for HLA-DR and PD-L1 expression on CD14^(+)monocytes using flow cytometry.Results:Compared with control subjects,the percentage of circulating CD14^(+)monocytes,HLA-DR+/CD14^(+)monocyte ratios,and mean fluorescence intensity were significantly decreased in patients with OHCA.After ROSC,HLA-DR expression in CD14^(+)monocytes in the TTM group was lower than that in patients with OHCA.However,there were no significant differences in the percentage of PD-L1+/CD14^(+)monocytes or the mean fluorescence intensity between patients with OHCA and healthy volunteers.Conclusion:After ROSC,circulating CD14^(+)monocytes and HLA-DR+/CD14^(+)monocyte ratios decreased significantly in patients with OHCA.Human leukocyte antigen D–related expression in CD14^(+)monocytes was lower in patients treated with TTM.

Onset of Coronary Heart Disease is Associated with HCMV Infection and Increased CD14^+CD16^+Monocytes in a Population of Weifang,China

Objective To investigate the relationship between human cytomegalovirus(HCMV)infection and peripheral blood CD14+CD16+monocytes in the pathogenesis of coronary heart disease(CHD),and to elucidate the mechanism of pathogenesis in CHD by analyzing the correlation between infection,inflammation,and CHD,to provide a basis for the prevention,evaluation,and treatment of the disease.Methods In total,192 patients with CHD were divided into three groups:latent CHD,angina pectoris,and myocardial infarction.HCMV-IgM and-IgG antibodies were assessed using ELISA;CD14+CD16+monocytes were counted using a five-type automated hematology analyzer;mononuclear cells were assessed using fluorescence-activated cell sorting;and an automatic biochemical analyzer was used to measure the levels of triglyceride,cholesterol,high-and low-density lipoprotein cholesterols,lipoprotein,hs-CRp and Hcy.Results The positive rates of HCMV-IgM and-IgG were significantly higher in the CHD groups than in the control group.HCMV infection affects lipid metabolism to promote immune and inflammatory responses.Conclusion HCMV infection has a specific correlation with the occurrence and development of CHD.The expression of CD14+CD16+mononuclear cells in the CHD group was increased accordingly and correlated with acute HCMV infection.Thus,HCMV antibody as well as peripheral blood CD14+CD16+mononuclear cells can be used to monitor the occurrence and development of CHD.

Caspase-1 activation and mature interleukin-1β release are uncoupled events in monocytes

AIM:To investigate whether caspase-1 activation/intracellular processing of pro-interleukin-1β(pro-IL-1β) and extracellular release of mature IL-1β from activated monocytes are separable events.METHODS:All experiments were performed on fresh or overnight cultured human peripheral blood monocytes(PBMCs) that were isolated from healthy donors.PBMCs were activated by lipopolysaccharide(LPS) stimulation before being treated with Adenosine triphosphate(ATP,1 mmol/L),human α-defensin-5(HD-5,50 μg/mL),and/or nigericin(Nig,30 μmol/L).For each experiment,the culture supernatants were collected separately from the cells.Cell lysates and supernatants were both subject to immunoprecipitation with anti-IL1β antibodies followed by western blot analysis with anti-caspase-1 and anti-IL-1β antibodies.RESULTS:We found that pro-IL-1β was processed to mature IL-1β in LPS-activated fresh and overnight cultured human monocytes in response to ATP stimulation.In the presence of HD-5,this release of IL-1β,but not the processing of pro-IL-1β to IL-1β,was completely inhibited.Similarly,in the presence of HD-5,the release of IL-1β,but not the processing of IL-1β,was significantly inhibited from LPS-activated monocytes stimulated with Nig.Finally,we treated LPS-activated monocytes with ATP and Nig and collected the supernatants.We found that both ATP and Nig stimulation could activate and release cleaved caspase-1 from the monocytes.Interestingly,and contrary to IL-1β processing and release,caspase-1 cleavage and release was not blocked by HD-5.All images are representative of three independent experiments.CONCLUSION:These data suggest that caspase-1 activation/processing of pro-IL-1β by caspase-1 and the release of mature IL-1β from human monocytes are distinct and separable events.

HCV-specific cytokine induction in monocytes of patients with different outcomes of hepatitis C

AIM:Cytokine release by macrophages critically determines the type of immune response to an antigen.Therefore,we studied hepatitis C virus(HCV)-specific induction of interleukins-1β,-10,-12(IL-1β,IL-10,IL-12),and tumor necrosis factor-α(TNF-α)in monocytes. METHODS:Intracallular cytokine expression was studied by flow cytometry in 23 patients with chronic hepatitis C,14 anti-HCV seropositives without viremia and 11 controls after stimulation of peripheral blood mononuclear calls with recombinant core,NS3,NS4,NSSa and NSSb proteins. RESULTS:Patients with HCV viremia revealed greater spontaneous expression of IL-1β,TNF-α,and IL-10. Furthermore,greater than twofold higher IL-10 expression was induced by the HCV antigens in chronic hepatitis C than in the other two groups(P<0.05).In contrast,neither IL- 12 nor TNF-α was induced preferentially. CONCLUSION:In chronic hepatitis C antigen-specific cytokine induction in monocytes is apparently shifted towards predominant IL-10 induction-not counterbalanced by antiviral type 1 cytokines.This may contribute to persistent viral replication.

Monocyte chemotactic protein-1 and soluble adhesion molecules as possible prognostic markers of the efficacy of antiviral treatment in chronic hepatitis C

AIM:To explain the role of Monocyte chemotactic protein-1 (MCP-1) and soluble adhesion molecules in chronic hepatitis C during the treatment of interferon alpha (IFNα) 2 b and ribavirin (RBV). METHODS:Concentrations of MCP-1,soluble adhesion molecules intercellular adhesion molecule-1 (sICAM-1),sP- selectin,interleukin (IL) 6,and IL10 in serum were estimated in the group of 40 patients with chronic hepatitis C treated with IFNalpha2 b and RBV in 0,16,32,48 wk of the therapy, RESULTS:In chronic hepatitis C,before and during the treatment,the serum levels of MCP-1 and sP-selectin in responders were similar to those of healthy subjects.In non- responders (NR),MCP-1 increased in the course of IFNc^+RBV treatment,differences were statistically significant as compared to responders.MCP-1 correlated statistically with the activity of periportal inflammation (r=0.35,P<0.05) but not with staging of liver fibrosis,sICAM-1 positively correlated with inflammatory activity and fibrosis in NR.sP-selectin did not correlate with histological findings in the liver.The MCP-1 correlated with the soluble form of sP-selectin concentrations (r= 6,P<0.001) and with IL-10 level in NR (r=0.4,P<0.05).There was no correlation observed between the concentration of MCP-1 and sICAM-1,IL-6 during the treatment. CONCLUSION:MCP-1 concentration may be a prognostic marker of the efficacy of IFN+RBV therapy in patients with chronic hepatitis C.

The First Reported Case of Human Monocytic Ehrlichiosis (HME) in Jordan

Human Ehrlichiosis infrequently occurs and can be missed, but attention to history and a meticulous physical examination would raise the index for suspicion and is documented with proper investigations. We report the first case of human monocytic Ehrlichiosis (HME) in a young female patient who lives in the Suburb city of Madaba, Jordan. She presented with fever, severe headache, skin rash, and confusion. She rapidly deteriorated and was admitted to our hospital. She had arrhythmias, convulsions, lapsed into a coma and respiratory failure and needed non-invasive ventilation. In addition to her clinical and epidemiological characteristics, the diagnosis was confirmed by the buffy coat. She had a swift response to oral doxycycline and was discharged home.

脓毒症合并2型糖尿病患者单核细胞人白细胞DR抗原的表达水平分析

目的观察2型糖尿病(T2DM)对脓毒症患者外周血单核细胞表面人白细胞DR抗原(HLA-DR)表达水平的影响。方法选取首都医科大学附属北京朝阳医院2021年3月至2022年2月收治的65例脓毒症合并T2DM(脓毒症合并T2DM组)、67例脓毒症未合并T2DM(脓毒症组)、60例T2DM患者(T2DM组)作为研究对象,以45例同期本院健康体检志愿者作为健康对照组。收集受试者性别、年龄、既往史、糖化血红蛋白(HbAlc)、入院时血糖、白细胞计数(WBC)淋巴细胞计数(LYM)超敏C-反应蛋白(hs-CRP)、主要感染部位,入院24h内进行序贯器官衰竭评分(SOFA)。采用流式细胞术检测外周血HLA-DR+CD14^(+)细胞表达百分比及平均荧光强度(MFI);并随访28d,观察T2DM对脓毒症患者28d预后的影响,比较不同预后患者HLA-DR^(+)CD14^(+)水平的差异。结果T2DM组HLA-DR^(+)CD14^(+)细胞表达百分比和MFI均明显低于健康对照组[HLA-DR^(+)CD14^(+)细胞表达百分比:87.72%(76.18%,93.64%)比94.86%(92.91%,95.70%),HLA-DR^(+)CD14^(+)MFI:10.80(8.45,14.45)比12.40(1.45,15.28)],脓毒症合并T2DM组和脓毒症组HLA-DRCD14^(+)细胞表达百分比和MFI均较健康对照组及T2DM组进一步降低[HLA-DR^(+)CD14^(+)细胞表达百分比:70.78%(42.22%,84.73%)、68.95%(44.95%,87.00%)比94.86%(92.91%,95.70%)、87.72%(76.18%,93.64%),HLA-DR^(+)CD14^(+)MFI:5.50(3.81,9.20)、5.29(3.35,9.59)比12.40(1.45,15.28)、10.80(8.45,14.45),均P<0.05]。脓毒症组和毒症合并T2DM组两组HLA-DR^(+)CD14^(+)细胞表达百分比和MFI、SOFA评分、28d病死率比较差异均无统计学意义(均P>0.05)。脓毒症组和脓毒症合并T2DM组死亡者的年龄明显大于生存者[岁:脓毒症组为68(60,74)比61(52,69),脓毒症合并T2DM组为66(64,73)比60(53,68),均P<0.05],SOFA评分明显高于生存者[分:脓毒症组为14(11,16)比8(5,11),脓毒症合并T2DM组为12(9,16)比8(6,11),均P<0.05],死亡组的HLA-DR^(+)CD14^(+)百分比和HLA-DR^(+)CD14^(+)MFI均明显低于生存组[HLA-DR^(+)CD14^(+)细胞表达百分比:脓毒症组为44.94%(28.01%,64.45%)比77.14%(47.41%,88.35%),脓毒症合并T2DM组为40.68%(34.83%,66.64%)比73.46%(58.44%,85.31%);HLA-DR^(+)CD14^(+)MFI:脓毒症组为3.92(2.30,5.44)比7.07(3.39,10.55),脓毒症合并T2DM组为3.90(3.34,6.04)比6.81(4.41,9.32),均P<0.05]。相关性分析显示:T2DM组、脓毒症组和脓毒症合并T2DM组HLA-DR^(+)CD14^(+)细胞表达百分比与hs-CRP均呈负相关(r值分别为-0.448、-0.628、-0.457,均P<0.001),MFI与hs-CRP亦呈负相关(r值分别为-0.289、-0.540、-0.323,均P<0.05)。脓毒症组和脓毒症合并T2DM组的HLA-DR^(+)CD14^(+)百分比与SOFA评分均呈负相关(r值分别为-0.520、-0.558,均P<0.001),MFI与SOFA评分也均呈负相关(r值分别为-0.327、-0.482,均P<0.01)。结论仑HLA-DR^(+)CD14^(+)水平在T2DM和脓毒症时均下降,但当脓毒症合并T2DM时,HLA-DR+CD14^(+)水平未发现进一步下降。

乌司他丁辅助治疗的脓毒症患者HLE、mCD14水平变化及早期检测HLE、mCD14水平对治疗反应性的评估价值

目的:探讨乌司他丁辅助治疗脓毒症时,患者血人白细胞弹性蛋白酶(HLE)、单核细胞表面脂多糖受体(mCD14)水平变化,以及早期检测血HLE、mCD14水平对治疗反应性的评估价值。方法:选取2020—2022年该院脓毒症患者196例,均在经典常规治疗方案基础上加用乌司他丁治疗,根据治疗效果分为反应良好组(38例)、反应差组(158例)。治疗前后检测两组患者血HLE、mCD14水平。通过电子病历系统收集患者资料,根据最邻近匹配法对两组进行1∶1匹配。分析乌司他丁辅助治疗脓毒症患者的治疗反应性影响因素并建立预测模型,通过受试者工作特征曲线(ROC曲线)分析早期检测血HLE、mCD14水平对治疗反应性的评估价值。结果:两组患者治疗后的血HLE水平低于治疗前,mCD14水平高于治疗前,差异均有统计学意义(P<0.05)。反应差组患者倾向性评分匹配后的严重程度和治疗前后降钙素原(PCT)水平、C反应蛋白(CRP)水平、HLE水平高于反应良好组,治疗前后mCD14水平低于反应良好组,差异均有统计学意义(P<0.05)。对于乌司他丁辅助治疗脓毒症患者,严重程度和治疗前PCT、CRP、HLE水平是治疗反应性的危险因素,治疗前mCD14水平是治疗反应性的保护因素(P<0.05);建立联合预测模型,P=1/[1+1/exp(0.826×严重程度+0.941×治疗前PCT水平+0.694×治疗前CRP水平+1.112×治疗前HLE水平-0.963×治疗前mCD14水平-3.968)],有较好的校准能力;治疗前HLE、mCD14水平预测治疗反应性的药时曲线下面积(AUC)分别为0.777、0.779,而联合预测模型预测治疗反应性的AUC为0.947,高于HLE、mCD14单独评估的AUC,差异有统计学意义(P<0.05)。结论:采用乌司他丁辅助治疗脓毒症,患者血HLE水平降低,mCD14水平升高;血HLE水平降低、mCD14水平升高与治疗反应性密切相关,联合检测可进一步提升预测价值。

康柏西普注射液对糖尿病视网膜病变患者的治疗效果

目的探讨康柏西普注射液对糖尿病视网膜病变(DR)患者血清人单核细胞趋化蛋白-1(MCP-1)、胎盘生长因子(PIGF)水平的影响。方法前瞻性研究,选取2020年5月至2022年8月西安市第一医院收治的193例DR患者作为研究对象,所有患者均采用康柏西普玻璃腔内注射液治疗。治疗1个月后,评估临床治疗效果,并分为疗效不佳组(27例)和疗效良好组(166例)。疗效不佳组男15例、女12例,年龄(53.14±9.45)岁,糖尿病病程(12.63±2.89)年;疗效良好组男86例、女80例,年龄(51.86±9.72)岁,糖尿病病程(10.82±3.51)年。对比两组患者临床资料及血清MCP-1、PIGF水平,采用logistic回归分析康柏西普注射液对DR患者治疗效果的危险因素,以及血清MCP-1、PIGF水平对DR患者治疗效果的预测效能。采用t检验、χ^(2)检验。结果疗效不佳组糖尿病病程、糖化血红蛋白(HbA1c)、血管内皮生长因子(VEGF)、MCP-1、PIGF水平均高于疗效良好组[(12.63±2.89)年比(10.82±3.51)年、(9.03±0.81)%比(8.52±0.94)%、(102.47±21.57)ng/L比(89.65±20.18)ng/L、(906.49±83.52)ng/L比(684.53±70.42)ng/L、(75.83±9.25)ng/L比(62.01±8.43)ng/L],差异均有统计学意义(t=2.541、2.662、3.032、14.786、7.793,均P<0.05)。logistic多因素回归分析显示,HbA1C、VEGF、MCP-1及PIGF水平均是影响DR患者康柏西普注射液治疗效果的危险因素(均P<0.05)。受试者操作特征曲线(ROC)分析结果显示,血清MCP-1、PIGF及二者联合对DR患者康柏西普注射液治疗效果预测的曲线下面积(AUC)分别为0.833、0.811、0.909。结论康柏西普注射液治疗可降低DR患者血清MCP-1、PIGF水平,且二者联合治疗效果的预测效能较高。

炎症相关标志物对初诊结肠癌疾病严重程度的预测作用

目的探讨炎症相关标志物对初诊结肠癌严重程度的预测作用。方法回顾2020年1月29日至2020年12月13日于山西省人民医院肛门结直肠中心住院并行手术治疗明确为结肠癌的患者,共收集212例患者的资料,寻找炎症相关标志物:外周血单核细胞/淋巴细胞比值(MLR)、外周血中性粒细胞/淋巴细胞比值(NLR)、和外周血血小板/淋巴细胞比值(PLR)与肿物大小、淋巴结转移、肝转移的关系、美国癌症联合会(AJCC)分期,以及与肿瘤标志物:癌胚抗原(CEA)、糖类抗原199(CA199)、糖类抗原CA125水平之间的关系。结果①初诊的212例结肠癌患者中男性、年龄>60岁占一半以上,肿物>5 cm患者炎症相关标志物MLR、NLR和PLR明显升高(P值均<0.05)。存在淋巴结转移患者炎症相关标志物MLR、NLR和PLR升高(P值均<0.05)。MLR、NLR和PLR数值在AJCC分期高的组高于AJCC分期组的低(P值均<0.05)。②炎症相关标志物(MLR、NLR和PLR)与肿瘤标志物具有相关性,MLR与CEA、CA199、CA125水平相关(P值均<0.05)。NLR与CEA、CA199、CA125水平相关(P值均<0.05)。PLR与CEA、CA199、CA125水平亦相关(P值均<0.05)。结论炎症标志物MLR、NLR和PLR组合可作为新诊断的结肠癌者肿瘤特征的工具。

重症肺炎患者外周血CD64指数、NLR及CD14^(+)单核细胞HLA-DR的检测意义分析

目的:探讨重症肺炎患者外周血CD64指数、中性粒细胞/淋巴细胞比值(NLR)及CD14^(+)单核细胞人类白细胞DR抗原(HLA-DR)的检测意义。方法:收集2022年1月—2024年3月抚州市第一人民医院收治的80例重症肺炎患者的临床资料进行研究。根据28 d内患者预后情况分组,将死亡患者作为死亡组(n=32),存活患者作为存活组(n=48)。比较两组一般资料及CD64指数、NLR、CD14^(+)单核细胞HLA-DR水平,分析CD64指数、NLR、CD14^(+)单核细胞HLA-DR单独及联合预测重症肺炎预后的价值,以及重症肺炎患者病死的危险因素。结果:两组年龄、入院时急性生理学和慢性健康状况评价Ⅱ(APACHEⅡ)比较,差异均有统计学意义(P<0.05)。死亡组CD64指数、NLR均高于存活组,CD14^(+)单核细胞HLA-DR水平低于存活组(P<0.05)。受试者操作特征(ROC)曲线结果显示,CD64指数、NLR、CD14^(+)单核细胞HLA-DR单独及联合预测重症肺炎预后不良的曲线下面积(AUC)分别为0.812、0.769、0.728、0.893,均具有一定预测价值(P<0.05)。logistic回归分析显示,CD64指数>9.15、NLR>12.19、CD14^(+)单核细胞HLA-DR≤44.60%、年龄≥65岁、入院时APACHEⅡ≥21.69分是重症肺炎患者病死的独立危险因素(P<0.05)。结论:CD14^(+)单核细胞HLA-DR水平偏低,CD64指数、NLR偏高提示重症肺炎患者预后不佳,三者联合检测可为临床预测重症肺炎患者预后提供参考。

cFLIPS regulates alternative NLRP3 inflammasome activation in human monocytes

The innate immune responses,including inflammasome activation,are paramount for host defense against pathogen infection.In contrast to canonical and noncanonical inflammasome activation,in this study,heat-killed gram-negative bacteria(HK bacteria)were identified as single-step stimulators of the NLRP3 inflammasome in human monocytes,and they caused a moderate amount of IL-1βto be released from cells.Time course experiments showed that this alternative inflammasome response was finished within a few hours.Further analysis showed that the intrinsically limited NLRP3 inflammasome activation response was due to the negative regulation of caspase-8 by the short isoform of cFLIP(cFLIPs),which was activated by NF-κB.In contrast,overexpressed cFLIPS,but not overexpressed cFLIPL,inhibited the activation of caspase-8 and the release of IL-1βin response to HK bacteria infection in human monocytes.Furthermore,we demonstrated that TAK1 activity mediated the expression of cFLIPs and was upstream and essential for the caspase-8 cleavage induced by HK bacteria in human monocytes.The functional specificity of cFLIPs and TAK1 revealed unique responses of human monocytes to a noninvasive pathogen,providing novel insights into an alternative regulatory pathway of NLRP3 inflammasome activation.

Bioinformatic prediction and functional characterization of human KIAA0100 gene

Our previous study demonstrated that human KIAA0100 gene was a novel acute monocytic leukemia-associated antigen (MLAA) gene. But the functional characterization of human KIAA0100 gene has remained unknown to date. Here, firstly, bioinformatic prediction of human KIAA0100 gene was carried out using online softwares; Secondly, Human KIAA0100 gene expression was downregulated by the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) 9 system in U937 cells. Cell proliferation and apoptosis were next evaluated in KIAA0100-knockdown U937 cells. The bioinformatic prediction showed that human KIAA0100 gene was located on 17q11.2, and human KIAA0100 protein was located in the secretory pathway. Besides, human KIAA0100 protein contained a signalpeptide, a transmembrane region, three types of secondary structures (alpha helix, extended strand, and random coil) , and four domains from mitochondrial protein 27 (FMP27). The observation on functional characterization of human KIAA0100 gene revealed that its downregulation inhibited cell proliferation, and promoted cell apoptosis in U937 cells. To summarize, these results suggest human KIAA0100 gene possibly comes within mitochondrial genome; moreover, it is a novel anti-apoptotic factor related to carcinogenesis or progression in acute monocytic leukemia, and may be a potential target for immunotherapy against acute monocytic leukemia.

Efficacy of recombinant human osteoprotegerin combined with tinidazole in the treatment of periodontitis mice and its correlation with serum RANKL and MCP-1 levels

Objective: To investigate the effect of recombinant human osteoprotegerin combined with tinidazole on mice with periodontitis and the effect on serum RANKL and MCP-1 levels. Methods: 80 SPF-cleaned mice were randomly divided into 4 groups, 20 each, model group, tinidazole group and recombinant human osteoprotegerin group were modeled by Kimura et al., and tinidazole group received tinidazole. After intragastric administration, the recombinant human osteoprotegerin group was injected with recombinant human osteoprotegerin in the periodontal pocket according to the tinidazole group. The periodontal changes of the four groups of mice were observed and recorded, and the gingival rating was performed. Epithelial tissue morphology was observed by hematoxylin-eosin (HE) staining. Serum levels of IL-4, IL-6, RANKL and MCP-1 were measured by enzyme-linked immunosorbent assay. Results:After the intervention, the model group developed severe inflammatory reactions, including redness, hemorrhage, and deep periodontal pockets. The teeth were significantly loosened. The mice in the tinidazole group and the recombinant human osteoprotegerin group recovered substantially, and the gingival rating of the recombinant human osteoprotegerin group was better than that. The tinidazole group and the model group (P<0.05). The results of HE staining showed that the model group had edema, vasodilation and a large amount of inflammatory infiltration. The epithelial structure of the mice in the tinidazole group and the recombinant human osteoprotegerin group was intact and arranged closely and orderly. After intervention, the IL-4 in the tinidazole group and the recombinant human osteoprotegerin group was significantly higher than the model group and IL-6 was significantly lower than the model group (P<0.05), and the recombinant human osteoprotegerin group IL-4 was significantly higher after the intervention. IL-6 was significantly lower in the tinidazole group than in the tinidazole group (P<0.05). After the intervention, the tinidazole group and the recombinant human osteoprotegerin group were significantly reduced, and the recombinant human osteoprotegerin group RAKNL and MCP-1 were significantly lower than the model group (P>0.05). Conclusion: Recombinant human osteoprotegerin combined with tinidazole has a better therapeutic effect on gums and teeth in mice with periodontitis, and can lower the levels of RAKNL and MCP-1 in serum, inhibit bone resorption and protect teeth.

Microarray Analysis of the Effects of Amelogenin on U937 Monocytic Cells

Periodontal diseases are chronic inflammation caused by particular types of bacteria and have been recognized as a cause of tooth loss in adults. These bacteria which invade periodontal tissue are phagocytosed mainly by monocytes and macrophages in this immune response, and will be presented to lymphocytes. Recently, therapies for regenerating periodontal tissues have been used extensively to treat periodontal disease, and in particular, enamel matrix derivative (EMD) is commonly used for such therapies in Japan. Amelogenin is a type of the extracellular matrix protein that accounts for 90% of the constituents of EMD. In this study, we carried out a detailed microarray analysis in order to evaluate a gene group involved in amelogenin stimuli in the human monocytic cell line U-937. Microarray analysis revealed that statistically significant changes were apparent in 273 genes (163 up-regulated and 110 down-regulated) subsequent to 4 h of amelogenin stimulation. The most highly enriched categories included “cell cycle”, “DNA replication”, and “DNA repair” in up-regulated annotation terms. On the other hand, “type I diabetes mellitus”, “allograft rejection”, and “graft versus host disease” were observed in down-regulated annotation terms. Specifically, the gene expression of major to compatibility complex (MHC) class I/II and CD80/86 was impaired in U937 cells after stimulation with amelogenin. In addition, the results of heat-map showed that the gene expression of inflammatory cytokine such as tumor necrosis factor (TFN), interleukin-18 (IL-18), and CXCL16 was markedly decreased after stimulation of monocytes with amelogenin. In conclusion, the findings of our study showed that by inducing monocyte growth through the suppression of the antigen-presenting ability of U937 cells, amelogenin may affect the immune responses of periodontal tissues originating from monocytes. Examining the effects of amelogenin on the transformation of macrophages differentiating from monocytes may establish a molecular basis for the anti-inflammatory effect of amelogenin in periodontal tissues.