慢性鼻窦炎伴鼻息肉患者嗅觉障碍的影响因素分析

目的分析慢性鼻窦炎伴鼻息肉(chronic rhinosinusitis with nasal polyps,CRSwNP)患者嗅觉障碍的影响因素。方法回顾性分析2014—2018年就诊于北京安贞医院行内镜鼻窦手术治疗的CRSwNP患者88例,男性22例,女性66例,年龄(48.1±11.3)岁(±s,下同)。所有入选患者均于术前行Sniffin′Sticks嗅觉测试、Lund-Mackay评分及改良鼻窦CT嗅区评分、鼻阻力及声反射检查、血常规及血生化等实验室检测、血清特异性IgE检测;术中取鼻息肉组织进行嗜酸粒细胞计数。根据Sniffin′Sticks嗅觉测试结果将患者分成嗅觉功能正常组和嗅觉功能障碍组,两组之间进行临床基线资料比较,根据单因素分析结果,结合临床有意义的指标进一步行多因素Logistic回归模型分析,并初步建立CRSwNP嗅觉障碍的预测模型。设P<0.05为差异有统计学意义。结果88例CRSwNP患者中,嗅觉正常32例(36.4%),嗅觉障碍56例(63.6%),其中嗅觉下降40例(45.5%)、失嗅16例(18.2%)。单因素分析发现,两组间组织嗜酸粒细胞数、血嗜酸粒细胞百分比、血尿素的差异存在统计学意义[12.7[2.0,52.3]个/高倍视野(M[P25,P75],下同)比38.6[16.2,87.0]个/高倍视野、2.75[1.60,4.80]%比4.35[2.50,6.60]%、(5.56±1.15)mmol/L比(4.98±1.33)mmol/L,P值均<0.05];改良鼻窦CT嗅区评分、除窦口鼻道复合体评分外的Lund-Mackay评分的差异均存在统计学意义(P值均<0.05)。多因素Logistic回归模型分析发现,改良鼻窦CT双侧嗅区总分和血尿素的差异具有统计学意义,其中双侧嗅区总分是嗅觉功能的危险因素(OR=2.108,95%CI:1.407~3.159,P<0.001);一定浓度的血尿素是嗅觉功能的保护因素(OR=0.461,95%CI:0.240~0.884,P=0.020)。进一步研究发现,由组织嗜酸粒细胞计数、血嗜酸粒细胞百分比、改良鼻窦CT双侧嗅区总分、总吸气、血尿素组成的预测模型受试者工作特征(ROC)曲线下面积(AUC)的值为0.888(P<0.01),对CRSwNP嗅觉障碍预测效果较好。结论改良鼻窦CT嗅区评分与CRSwNP患者的嗅觉障碍密切相关,一定程度的血尿素升高可能对CRSwNP患者的嗅觉功能有保护作用。     

Interleukin-3, -5, and Granulocyte Macrophage Colony-Stimulating Factor Induce Adhesion and Chemotaxis of Human Eosinophils via p38 Mitogen-Activated Protein Kinase and Nuclear Factor κB

Hematopoietic cytokines such as interleukin (IL)-3, IL-5, and granulocyte macrophage colony-stimulating factor (GM-CSF) play a fundamental role in eosinophil functions in allergic asthma. The intracellular signal transduction mechanisms of these cytokines regulating the activation of eosinophils have been potential therapeutic targets. We investigated the roles of p38 mitogen-activated protein kinase (MAPK) and nuclear factor kappa-B (NF-κB) in IL-3, IL-5, and GM-CSF-induced adhesion, morphological changes, and subsequence transmigration of human eosinophils. IL-3, IL-5, and GM-CSF could augment the phosphorylation of p38 MAPK and nucleus translocation of NF-κB in eosinophils. cDNA expression arrays demonstrated that the gene expression levels of several adhesion molecules including intercellular adhesion molecule-1 (ICAM-1), α6, β2 integrin (CD18), and CD44 were upregulated by these cytokines. Results from functional assays showed that adhesion of eosinophils onto airway epithelial cells was enhanced after IL-3 and IL-5 but not GM-CSF stimulation. These cytokines could markedly induce shape change and augment the transmigration of eosinophils. Moreover, administration of either p38 MAPK inhibitor, SB 203580, or proteasome inhibitor, N-cbz-Leu-Leu-leucinal (MG-132), could inhibit the cytokine-induced adhesion, shape change, and transmigration of eosinophils. Together, our findings suggest that IL-3, IL-5, and GM-CSF regulated the adhesion and chemotaxis of human eosinophils through shared signaling pathways involving both p38 MAPK and NF-κB. Our results therefore shed light on the further development of more effective agents for allergic and inflammatory diseases.     

The effect of pharmacological PI3Kγ inhibitor on eotaxin-induced human eosinophil functions

BackgroundAsthma is characterized by chronic inflammation caused by activation of immune cells including Th2 lymphocytes and eosinophils. Phosphoinositide 3-kinase (PI3K) γ deficient asthmatic mice did not develop lung eosinophilia, although the detailed mechanisms are not well known. A CC chemokine eotaxin (CCL11) plays a prominent role in developing eosinophilic inflammation through CCR3. In this study, we tested the roles of PI3Kγ in eotaxin-induced eosinophil functions using a pharmacological inhibitor.MethodHuman peripheral blood eosinophils were isolated by CD16-negative selection method. The effect of AS605240, synthetic PI3Kγ inhibitor on eotaxin-induced adhesion, chemotaxis, and degranulation were studied using intracellular adhesion molecule-1 (ICAM-1)-coated plates, Boyden chamber system, ELISA for eosinophil-derived neurotoxin (EDN) levels in the culture supernatant, respectively. CCR3 expression levels and extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation were assessed by flowcytometry. Involvement of PI3Kγ in spontaneous apoptosis was studied using flowcytometry.ResultsAlthough AS605240 did not affect the eosinophil spontaneous apoptosis, eotaxin-induced chemotaxis, adhesion to ICAM-1 coated plate, and EDN release were inhibited by AS605240. AS605240 also inhibited the eotaxin-induced ERK1/2 phosphorylation without down-regulation of surface CCR3 expression.ConclusionThese results indicate that PI3Kγ inhibitor attenuates eotaxin-induced eosinophil functions by suppressing the downstream signaling of CCR3 without significant cytotoxicity. PI3Kγ plays an important role in the development of eosinophilic inflammation and blockade of PI3Kγ might be a therapeutic strategy for treatment of eosinophil-related diseases including asthma.     

Eosinophil associated genes in the inflammatory bowel disease 4 region: Correlation to inflammatory bowel disease revealed

AIM: To study the association between inflammatory bowel disease (IBD) and genetic variations in eosinophil protein X (EPX) and eosinophil cationic protein (ECP).METHODS: DNA was extracted from ethylene diamine tetraacetic acid blood of 587 patients with Crohn’s disease (CD), 592 with ulcerative colitis (UC) and 300 healthy subjects. The EPX405 (G > C, rs2013109), ECP434 (G > C, rs2073342) and ECP562 (G > C, rs2233860) gene polymorphisms were analysed, by the 5’-nuclease allelic discrimination assay. For determination of intracellular content of EPX and ECP in granulocytes, 39 blood samples was collected and extracted with a buffer containing cetyltrimethylammonium bromide. The intracellular content of EPX was analysed using an enzyme-linked immunosorbent assay. The intracellular content of ECP was analysed with the UniCAP^® system as described by the manufacturer. Statistical tests for calculations of results were χ² test, Fisher’s exact test, ANOVA, Student-Newman-Keuls test, and Kaplan-Meier survival curve with Log-rank test for trend, the probability values of P < 0.05 were considered statistically significant.RESULTS: The genotype frequency for males with UC and with an age of disease onset of ≥ 45 years (n = 57) was for ECP434 and ECP562, GG = 37%, GC = 60%, CC = 4% and GG = 51%, GC = 49%, CC = 0% respectively. This was significantly different from the healthy subject’s genotype frequencies of ECP434 (GG = 57%, GC = 38%, CC = 5%; P = 0.010) and ECP562 (GG = 68%, GC = 29%,CC = 3%; P = 0.009). The genotype frequencies for females, with an age of disease onset of ≥ 45 years with CD (n = 62), was for the ECP434 and ECP562 genotypes GG = 37%, GC = 52%, CC = 11% and GG = 48%, GC = 47% and CC = 5% respectively. This was also statistically different from healthy controls for both ECP434 (P = 0.010) and ECP562 (P = 0.013). The intracellular protein concentration of EPX and ECP was calculated in μg/10⁶ eosinophils and then correlated to the EPX 405 genotypes. The protein content of EPX was highest in the patients with the CC genotype of EPX405 (GG = 4.65, GC = 5.93, and CC = 6.57) and for ECP in the patients with the GG genotype of EPX405 (GG = 2.70, GC = 2.47 and CC = 1.90). ANOVA test demonstrated a difference in intracellular protein content for EPX (P = 0.009) and ECP (P = 0.022). The age of disease onset was linked to haplotypes of the EPX405, ECP434 and ECP562 genotypes. Kaplan Maier curve showed a difference between haplotype distributions for the females with CD (P = 0.003). The highest age of disease onset was seen in females with the EPX405CC, ECP434GC, ECP562CC haplotype (34 years) and the lowest in females with the EPX405GC, ECP434GC, ECP562GG haplotype (21 years). For males with UC there was also a difference between the highest and lowest age of the disease onset (EPX405CC, ECP434CC, ECP562CC, mean 24 years vs EPX405GC, ECP434GC, ECP562GG, mean 34 years, P = 0.0009). The relative risk for UC patients with ECP434 or ECP562-GC/CC genotypes to develop dysplasia/cancer was 2.5 (95%CI: 1.2-5.4, P = 0.01) and 2.5 (95%CI: 1.1-5.4, P = 0.02) respectively, compared to patients carrying the GG-genotypes.CONCLUSION: Polymorphisms of EPX and ECP are associated to IBD in an age and gender dependent manner, suggesting an essential role of eosinophils in the pathophysiology of IBD.     

Galectin-9-like from Angiostrongylus cantonensis young adult worms modulates eosinophil chemotaxis in vitro

Background/purposeEosinophils are recruited to the brain of mice after infection with Angiostrongylus canonensis. Several factors produced by infected mice are well known playing the role to chemoattract eosinophils from the blood into the brain. The purpose of this study is to investigate whether Angiostronylus cantonensis young-adult worms (AcYA) have components which have eosinophilic chemotactic activity.MethodsEosinophil chemotactic activity of AcYA was tested by Boyden blind-well chamber technique. The components of AcYA were analysed by SDS-PAGE and Mass spectrometry. Furthermore, galectin-9 in the cerebrospinal fluid (CSF) of infected mice and galectin-9-like in AcYA were measured by ELISA technic and also were recognized by western blot analysis respectively.ResultsExcretory-secretory products of AcYA did not show eosinophil chemotactic activity. However, the extracts of AcYA showed protein concentration-dependent eosinophil chemotactic activity and reached the peak at the 24 μg/ml. The eosinophil chemotactic activity was significantly reduced by lactose. The components of AcYA at molecular weights of approximatively 15 kDa and 35 kDa showed several galectins component in Mass spectrometric analysis. Furthermore, galectin-9-like in AcYA was recognized by ELISA and western blot analysis. In parallel with increase of galectin-9 in the CSF, eosinophils were also significantly increased in mouse after infected with A. cantonensis.ConclusionGalectin-9-like in AcYA and galectin-9 in mouse CSF were confirmed demonstrating eosinophil chemotactic activity both in vitro study and in the infection of mouse in this study.     

Evaluation of plasma eosinophil count and mean platelet volume in patients with coronary slow flow

OBJECTIVE:

The pathophysiology of coronary slow flow has not been clearly defined, although multiple abnormalities including arteritis, endothelial dysfunction, and atherothrombosis, have been reported. It is known that eosinophils play an important role in inflammation, endothelial dysfunction, and thrombosis. We aimed to compare the eosinophil counts of coronary slow flow patients versus healthy controls.

METHODS:

This study included 50 coronary slow flow patients (19 males, mean age 65.6±13.7 years) and 30 healthy controls (10 males, mean age 57.86±11.6 years). These participants were evaluated using concurrent routine biochemical tests as well as neutrophil, lymphocyte, and eosinophil counts and mean platelet volume (MPV), which were obtained from the whole blood count. These parameters were compared between groups.

RESULTS:

The baseline characteristics of the study groups were comparable. The coronary slow flow patients had a higher mean platelet volume and eosinophil count than the control group (8.38±0.86 vs 6.28±1.6 fL and 0.31±0.42 vs 0.09±0.05; p<0.001 and 0.008, respectively).

CONCLUSION:

Our study demonstrated a relationship between eosinophil count and MPV in patients with coronary slow flow.     

哮喘致死患者肺组织EG1和EG2的检测及分析

目的　为了研究哮喘发作致死患者肺组织嗜酸性粒细胞浸润情况。方法　研究分成 3组 ,A组为死于哮喘发作 ,B组有哮喘病史但死于非哮喘发作 ,C组无哮喘病史。应用免疫组化技术 ,检测不同死因肺组织单克隆抗体EG1 和EG2 的阳性细胞。结果　 ( 1)哮喘发作致死组的肺组织EG1 和EG2 的阳性细胞明显高于其他两组 ,而哮喘患者非哮喘发作致死组的肺组织EG1 和EG2 的阳性细胞与无哮喘病史组比较无显著差异。 ( 2 )在哮喘发作致死组的支气管与肺实质EG1 阳性细胞数无明显差异 ,但在肺实质的EG2 的阳性细胞明显高于较大的支气管。结论　在哮喘发作致死患者气道和肺组织有大量的嗜酸性粒细胞浸润 ,且以小支气管与肺实质浸润为主     

变应性皮炎患者血清中可溶性E-选择素及总IgE检测

目的探讨变应性皮炎血清标志物间的相互关系.方法应用酶联免疫吸附测定法检测了20例变应性皮炎、18例荨麻疹患者及20例正常人对照血清中可溶性E-选择素,同时应用不同方法测定了其血清总免疫球蛋白E及嗜酸细胞阳离子蛋白.结果变应性皮炎患者血清中可溶性E-选择素及总免疫球蛋白E水平非常显著高于对照组(P＜0.001),且可溶性E-选择素与总免疫球蛋白E间具有显著相关性(γ=0.068,P=0.003);变应性皮炎患者血清中嗜酸细胞阳离子蛋白也有升高.上述血清标志物水平在治疗后降低.结论可溶性E-选择素可能是变应性皮炎严重性及活动性的良好标志物.