哮喘大鼠嗜铬蛋白A表达的检测及意义

目的探讨嗜铬蛋白A在哮喘发病机制中的作用。方法通过哮喘大鼠模型,分别采用免疫组化法和实时荧光定量PCR法检测肺组织和血中性粒细胞(PMN)嗜铬蛋白A或mRNA的表达。结果哮喘组肺组织嗜铬蛋白A及mRNA的表达水平显著高于对照组,差异有统计学意义(P0.01);布地奈德组嗜铬蛋白A及mRNA的表达水平显著低于哮喘组,差异有统计学意义(P0.01)。哮喘组血PMN嗜铬蛋白A mRNA的表达量显著高于对照组,差异有统计学意义(P0.01),布地奈德组嗜铬蛋白A mRNA的表达量与哮喘组差异无统计学意义(P0.05)。结论哮喘大鼠嗜铬蛋白A的表达升高,嗜铬蛋白A可能参与了哮喘的炎症机制,其表达能被布地奈德所抑制。     

重症肺炎患儿中性粒细胞CD11b的表达意义

目的:探讨中性粒细胞CD11b在重症肺炎儿童中的表达意义。方法:采用流式细胞术检测48例重症肺炎患儿和46例普通肺炎患儿急性期、恢复期以及47例正常儿童血清中性粒细胞CD11b表达水平。结果:急性期重症肺炎组和普通肺炎组中性粒细胞CD11b表达均高于正常对照组,差异有统计学意义(P<0.05);急性期重症肺炎组中性粒细胞CD11b表达要高于普通肺炎组,差异有统计学意义(P<0.05)。恢复期重症肺炎组和普通肺炎组中性粒细胞CD11b表达均低于同组急性期,差异有统计学意义(P<0.05)。重症细菌性肺炎组、重症病毒性肺炎组、重症支原体肺炎组、重症病毒与细菌混合感染性肺炎组急性期中性粒细胞CD11b的表达水平比较,差异均无统计学意义(P>0.05)。重症社区获得性肺炎组和重症院内获得性肺炎组急性期中性粒细胞CD11b的表达水平比较,差异无统计学意义(P>0.05)。结论:中性粒细胞CD11b的表达可能在儿童重症肺炎的演变过程中起到重要的作用。     

中性粒细胞在子痫前期发病中的作用

目的:明确中性粒细胞在子痫前期患者中的状态,了解中性粒细胞与子痫前期发病的关系。方法:采用流式细胞术检测20例子痫前期患者和20例正常妊娠孕妇肘静脉血中性粒细胞黏附分子CD11b的表达,同时采用酶联免疫吸附实验检测两组孕妇肘静脉血浆中性粒细胞的趋化因子IL-8和中性粒细胞释放的弹性蛋白酶的浓度。结果:①子痫前期组CD11b的平均对数荧光强度指数、血浆IL-8和弹性蛋白酶的浓度〔分别为(19.519±9.281)、(23.665±6.017)、(148.475±97.227)pg/ml〕均高于正常妊娠组〔分别为(13.871±6.872)、(18.831±3.472)、(87.102±58.545)pg/ml〕;②子痫前期孕妇的中性粒细胞CD11b平均对数荧光强度指数、血浆IL-8和弹性蛋白酶的浓度与中性粒细胞绝对数,入院血压、新生儿的出生体重无相关关系。结论:子痫前期患者中性粒细胞CD11b的表达增强,趋化因子IL-8浓度及中性粒细胞释放的弹性蛋白酶浓度升高,表明中性粒细胞在子痫前期中活性增强,可能与内皮细胞的损伤有关,在子痫前期的发病中起重要作用。     

中性粒细胞CD162、CXCR2、CXCR4表达与小儿肺炎支原体感染的关系

目的 探究中性粒细胞P-选择素糖蛋白配体1(PSGL-1/CD162)、趋化因子受体-2(CXCR2)、趋化因子受体-4(CXCR4)的表达与小儿肺炎支原体(MP)感染的关系。方法 选取2020年3月-2021年3月海南省妇女儿童医学中心接收的78例MP感染患儿为感染组,其中重症36例、轻症42例,以同期来院体检的健康儿童40名为对照组,流式细胞术和实时荧光定量聚合酶链式反应(qRT-PCR)检测中性粒细胞CD162、CXCR2、CXCR4的表达水平。结果 感染组患儿外周血中性粒细胞CD162、CXCR2、CXCR4的平均荧光强度(MFI)分别为(87.33±13.34)、(1.33±0.28)、(1.64±0.31),mRNA相对表达量分别为(2.36±1.26)、(3.65±0.64)、(4.33±1.18)均高于对照组(P<0.05);重症MP感染患儿高于轻症患儿(P<0.05)。受试者工作特征曲线(ROC)分析显示,中性粒细胞CD162、CXCR2和CXCR4的MFI水平联合评估MP重症感染的曲线下面积为0.881(95%CI:0.801～0.961),mRNA含量...     

剔除嗜中性粒细胞减少SD大鼠脊髓损伤诱导的急性肾损伤

目的评价嗜中性粒细胞的剔除减少SD大鼠脊髓损伤诱导肾功能损伤的作用。方法抗嗜中性粒细胞抗体处理SD大鼠24h后,对照组(A)仅切除椎板、抗血清未处理组(B)和抗血清处理组(C)行脊髓损伤术。术后24h,观察大鼠肾功能以及肾脏病变情况。结果术后24h,C组肌酐(Scr)、尿素氮(BUN)和丙二醛(MAD)水平显著降低,与B组比较,在统计学意义上有显著性差异(P<0.001),B组肾脏为典型大白肾,HE和PAS染色观察发现急性肾小管坏死症状,并表现炎症细胞浸润。结论剔除嗜中性粒细胞可以有效地减少大鼠脊髓损伤引起的肾组织病变从而有效地保护肾功能。     

中性粒细胞群落参数在术后并发感染时的变化

目的 分析中性粒细胞群落参数(VCS参数)在机体应激状态时的变化,探讨其诊断应激状态并发细菌感染的可能.方法 使用Coulter LH 750血液分析仪,检测41例经细菌培养确诊并发术后感染者和83例术后无感染症状者及212例正常对照者的外周血,记录白细胞计数(WBC)、中性粒细胞(NE)和中性粒细胞的各项VCS参数,并根据感染病原菌的革兰染色结果,将感染组分为革兰阴性菌感染组和革兰阳性菌感染组;然后再进行指标的组间比较分析.结果 无感染症状组WBC为(15.40±2.40)× 109/L,NE为(84.50±3.10)％,感染组WBC为(16.20±3.04)×109/L,NE为(85.60±3.40)％,两组差异无统计学意义;而中性粒细胞细胞平均体积(NEV)及其分布宽度(NVW)术后明显高于术前(P＜0.01);NEV、NVW在ROC曲线中的线下面积为0.800和0.777,要大于其他指标,在取临界值≥152.15和≥22.55时,其敏感性和特异性分别为70.73％、79.52％和90.24％、51.81％;在革兰阴性菌、革兰阳性菌感染组间,NEV和NEC差异有统计学意义(P＜0.05).结论 继发的细菌感染引起了应激状态下中性粒细胞VCS参数的显著性变化,NEV、NVW出现了显著性的增大,其对临床的诊治有一定帮助.     

重症监护病房患者中性粒细胞VCS参数检测的临床意义

目的研究重症监护病房患者感染后中性粒细胞细胞体积(V)、电导率(C)、激光散射(S)参数的变化。方法利用Beckman-Coulter LH750血球仪分别检测98例重症监护病房患者和40例对照组白细胞(WBC)、中性粒细胞(ANC)及其VCS参数:中性粒细胞平均体积(MNV)、中性粒细胞体积分布宽度(NDW)、中性粒细胞平均传导率(MNC)、中性粒细胞平均散射值(MNS),将98例患者根据微生物鉴定结果分为感染组和非感染组。结果感染组患者WBC(12.33±5.39)×109/L、ANC(10.04±5.17)×109/L及其VCS(MNV:157±18,NDW:24.38±4.18,MNC:136±12,MNS:137±11)参数与非感染组〔WBC:(8.71±4.22)×109/L,ANC:(6.79±4.50)×109/L,MNV:151±13,NDW:20.47±3.29,MNC:140±11,MNS:141±12〕、对照组〔WBC:(5.87±3.68)×109/L,ANC:(3.89±2.64)×109/L,MNV:144±8,NDW:18.61±1.39,MNC:144±8,MNS:145±9〕比较差异有统计学意义;非感染组患者WBC、ANC及其MNV、NDW参数与对照组比较差异有学意义;感染组患者经治疗后,细菌再培养阴性组VCS参数MNV(151±15,145±11)、NDW(22.48±3.65,19.31±3.29)与阳性组差异有统计学意义;MNV、NDW参数在感染诊断中要明显优于白细胞、中性粒细胞。结论中性粒细胞VCS参数是反应患者细菌感染的敏感指标。     

脊柱术后感染病原菌及CD64和中性粒细胞百分比的早期预测价值

目的分析脊柱术后医院感染患者病原菌分布特征及中性粒细胞64(CD64)和中性粒细胞百分比(NEU%)表达情况。方法选取2017年6月-2019年6月天津市天津医院收治的脊柱术后医院感染患者51例为感染组,并选取同期于医院就诊的脊柱术后未发生感染患者51例为未感染组。分析感染组患者病原菌情况,检测患者NEU和CD64水平;ROC曲线分析CD64和NEU%指标水平预测感染的诊断价值。结果 51例感染患者共培养分离病原菌78株,其中革兰阴性菌42株(53.85%)、革兰阳性菌33株(42.31%)、真菌3株(3.85%),以铜绿假单胞菌及金黄色葡萄球菌为主。术后7 d,感染组体温、WBC、中性粒细胞计数、CRP、ESR分别为(38.47±0.52)℃、(8.34±2.17)×109/L、(5.98±1.94)×109/L、(54.52±19.93)mg/L、(51.74±21.88)mm/h均高于非感染组(P<0.05)。术后7 d,感染组CD64、NEU%分别为(8.87±2.13)%、(84.93±13.39)高于非感染组(P<0.05)。CD64指数与NEU%水平诊断ROC曲线下面积分别为0.922、0.826。结论脊柱术后医院感染以革兰阴性菌为主,CD64及NEU%水平有助于脊柱术后医院感染的早期诊断。     

布地奈德对哮喘大鼠肺组织病理改变及TRAF2表达的影响

【目的】 观察哮喘大鼠肺组织病理改变和TRAF2表达的水平,探讨布地奈德治疗哮喘的可能作用机制。 【方法】 采用大鼠哮喘模型,随机分成哮喘组、对照组和布地奈德组,HE染色观察肺组织炎性改变,免疫组织化学法检测肺组织TRAF2的表达。 【结果】 哮喘组大鼠的毛发、体重、活动度等一般情况改变和肺组织炎性改变较对照组有显著变化,布地奈德能明显减轻上述改变。哮喘组(0.317±0.041 OD值)支气管壁TRAF2的光密度值显著高于对照组(0.220±0.057 OD值)(P＜0.01);布地奈德组支气管壁(0.236±0.033 OD值)TRAF2的光密度值显著低于哮喘组(P＜0.01),与对照组相比差异无统计学意义(P>0.05),但肺组织中其表达水平却显著高于哮喘组。 【结论】 哮喘大鼠TRAF2的表达水平增强,它可能参与了哮喘的气道炎症过程;布地奈德能减轻气道炎症,其机制可能是部分通过TRAF2途经。     

支气管哮喘患儿中性粒细胞胞外诱捕网水平与患者免疫功能及临床预后的关系

目的探讨支气管哮喘患儿中性粒细胞胞外诱捕网水平与患者免疫功能及临床预后的关系。方法选取本院2017年9月-2018年9月支气管哮喘患者80例作为观察组,并以同期健康体检的30例儿童作为对照组,采用pico Green dsDNA荧光染色定量检测支气管哮喘患者NETs水平,采用T淋巴细胞亚群评价患者免疫功能,对比不同NETs水平患者免疫功能及临床特点,采用Logistic回归分析探讨小儿支气管哮喘急性发作的风险因素。结果观察组白细胞、中性粒细胞、单核细胞、Cf-DNA/NETs水平均高于对照组(P <0. 05),而淋巴细胞水平低于对照组,差异均有统计学意义(P <0. 05)。Logistic回归分析显示,哮喘家族史、呼吸道感染、Cf-DNA/NETs水平上升、有害气体接触、被动吸烟均是支气管哮喘急性发作的独立危险因素(P <0. 05)。结论中性粒细胞胞外诱捕网水平与支气管哮喘患者免疫功能及预后密切相关,值得临床重点关注。     

乌司他丁对体外循环下中性粒细胞凋亡与呼吸爆发的影响

目的观察体外循环（CPB）心脏手术患者中性粒细胞（PMN）凋亡、呼吸爆发的变化及乌司他丁对其影响。方法选择在CPB下行瓣膜置换术患者62例,随机分成乌司他丁组（U组）和对照组（c组）,每组各31例。U组患者于麻醉诱导后给予乌司他丁,C组患者则给予等容积的0．9％NaCl溶液。分别于麻醉后手术前（T1）、CPB开始后30min（T2）、CPB停止后30min（T3）抽取动脉血,分离PMN,检测PMN凋亡率,呼吸爆发以及血浆中超氧化物歧化酶（SOD）和丙二醛（MDA）的水平。结果C组PMN凋亡率在T1为（66．57±5．93）％,T2为（55．37±3．51）％,T,为（48．92±4．21）％,T2、T1均较T1显著降低（P〈0．05）,并于L达最低值;U组PMN凋亡率在T。为（73．57±7．94）％,T2为（68．34±4．92）％,T3为（62．13±4．76）％,T2、L均较T1显著降低（P〈0．05）,并于T3达最低值;C组PMN凋亡率显著低于U组（P〈0．05）。两组PMN呼吸爆发均表现为CPB开始后逐渐升高,L达到峰值;U组T2（1105．94±84．15）MCF,T3（1156．52±93．20）MCF,与C组T2（1266．06±99．55）MCF,T3（1422．50±89．75）MCF比较明显降低（P〈0．05）。两组SOD均于手术开始后逐渐下降（P〈0．05）,C组T3SOD为（47．39±6．07）μU／L显著低于U组的（51．35±6．22）μU／L（P〈0．05）。两组MDA均于手术开始后逐渐升高（P〈0．05）,L达高峰,C组为（13．72±1．15）μmol／L,U组为（8．40±0．88）μmol／L,C组显著高于U组（P〈0．05）。结论CPB引起PMN凋亡率降低、凋亡延迟,呼吸爆发增强,乌司他丁能有效地促进过度激活的PMN凋亡、抑制PMN呼吸爆发,提高SOD,降低MDA,减轻CPB对机体的炎性反应及氧化损伤,对机体具有保护作用。     

乌司他丁对体外循环下中性粒细胞凋亡与呼吸爆发的影响

目的 观察体外循环(CPB)心脏手术患者中性粒细胞(PMN)凋亡、呼吸爆发的变化及乌司他丁对其影响.方法 选择在CPB下行瓣膜置换术患者62例,随机分成乌司他丁组(U组)和对照组(C组),每组各31例.U组患者于麻醉诱导后给予乌司他丁,C组患者则给予等容积的0.9%NaCI溶液.分别于麻醉后手术前(T1)、CPB开始后30 min(T2)、CPB停止后30 min(T3)抽取动脉血,分离PMN,检测PMN凋亡率,呼吸爆发以及血浆中超氧化物歧化酶(SOD)和丙二醛(MDA)的水平.结果 C组PMN凋亡率在T1为(66.57±5.93)%,T2为(55.37±3.51)%,T3为(48.92±4.21)%,T2、T3均较T1显著降低(P<0.05),并于T3达最低值;U组PMN凋亡率在T1为(73.57±7.94)%,T2为(68.34±4.92)%,T3为(62.13±4.76)%,T2、T3均较T1显著降低(P<0.05),并于T3达最低值;C组PMN凋亡率显著低于U组(P<0.05).两组PMN呼吸爆发均表现为CPB开始后逐渐升高,T3达到峰值;U组T2(1105.94±84.15)MCF,T3(1156.52±93.20)MCF,与C组T2(1266.06±99.55)MCF,T3(1422.50±89.75)MCF比较明显降低(P<0.05).两组SOD均于手术开始后逐渐下降(P<0.05),C组T3SOD为(47.39±6.07)μU/L显著低于U组的(51.35±6.22)μU/L(P<0.05).两组MDA均于手术开始后逐渐升高(P<0.05),T3达高峰,C组为(13.72±1.15)μmol/L,U组为(8.40±0.88)μmoI/L,C组显著高于U组(P<0.05).结论 CPB引起PMN凋亡率降低、凋亡延迟,呼吸爆发增强,乌司他丁能有效地促进过度激活的PMN凋亡、抑制PMN呼吸爆发,提高SOD,降低MDA,减轻CPB对机体的炎性反应及氧化损伤,对机体具有保护作用.     

Diurnal variation in neutrophil function

Neutrophil functions, including chemotaxis, reactive oxygen species (ROS)-producing capacity of neutrophils, and serum opsonic activity were investigated in 9 young healthy male volunteers. Venous blood of these volunteers was obtained under standardized conditions at 4-h intervals over a 24-h span. Neutrophil chemotaxis was evaluated by a modified Boyden technique, ROS-producing capacity of neutrophils and serum opsonic activity were measured by a simultaneous multiple measurement system based on luminol-dependent chemiluminescence and indicated by peak height and peak time. ROS-producing capacity of neutrophils and serum opsonic activity were activated in the daytime, and decreased from night to morning. There were negative correlations between the peak time of the luminol-dependent chemiluminescent response, neutrophil number (p<0.01) and segmented neutrophil number (p>0.01). On the other hand, no significant correlations were noted between serum opsonic activity and IgG, IgA, IgM, C3 or C4. In contrast, the peaks of neutrophil chemotaxis were at the wake-up time (6:00a.m.) and in the evening (6:00p.m.). This study indicates that diurnal variation of neutrophil function exists.     

中性粒细胞胞外诱捕网蛋白质的免疫原性

中性粒细胞胞外诱捕网(NETs)由DNA和抗菌蛋白质组成。在活细胞内,这些组分分布于亚细胞并发挥不同的功能。但是,在NETosis期,这些组分重新分布并从中性粒细胞内挤压出来。NETosis过分强盛和NETs清除受损,与自身免疫性疾病的器官损害有关,如系统性红斑狼疮(SLE)、类风湿性关节炎(RA)、Felty's综合征(FS)和小血管炎(SVV)等。NETs可能是体内新抗原的重要来源,蛋白质翻译后的修饰与水解,或药物诱导NETs构象异常能促进自身免疫性疾病患者产生自身抗体。总之,NETs可能提供了一个独特的具有破坏正常免疫耐受,引起自身免疫性的微环境。     

抗中性粒细胞胞外诱捕网的细菌毒力因子

中性粒细胞胞外诱捕网(Neutrophil extracellular traps,NETs)作为人体先天免疫系统的组成部分,具有捕杀病原体的作用。最近已经明确,几种细菌毒力因子有利于细菌逃避NETs的作用。本文讨论5种毒力因子,包括核酸酶、多糖荚膜、丙氨酰脂磷壁酸、唾液酸化聚糖分子模拟物、β-羟基丁酸。     

Antimalarial drugs and human neutrophil oxidative metabolism

The effect of several commonly used antimalarial drugs on human peripheral blood neutrophil oxidative metabolism was studied. The following drugs were tested: chloroquine diphosphate, quinine HCl, mefloquine, proguanil HCl, cycloguanil, pyrimethamine, sulphadoxine, and tetracycline HCl. It was found that none of the antimalarial drugs examined, at clinically obtainable concentrations, had any inhibitory effect on neutrophil oxygen consumption, glucose oxidation, superoxide production, NBT reduction, and chemiluminescence. However, at higher concentrations chloroquine, quinine, mefloquine, and proguanil inhibited neutrophil oxidative burst. There was a slight enhancing effect on neutrophil oxidative metabolism by pyrimethamine, combination of pyrimethamine-sulphadoxine, cycloguanil and tetracycline at concentrations lower than the clinical levels.     

Intravenous fat emulsion acutely suppresses neutrophil chemiluminescence

The immediate effect of intravenous fat emulsion on neutrophil oxidant release was studied. Opsonized nonencapsulated S. aureus was used to stimulate neutrophil activity. Luminol enhanced chemiluminescence was followed over 15 min and recorded as peak output (P; mV), integral under the curve (I; V-sec) and rate of increase (R; mV/sec). Eighteen chronically ill patients receiving glucose based total parenteral nutrition were studied before and after a 4- to 6-hr test infusion of 500 ml of 10% fat emulsion. P decreased from 719 +/- 46 to 461 +/- 42 mV (p less than 0.001), I decreased from 169 +/- 17 to 111 +/- 12 V-sec (p less than 0.001) and R decreased from 6.9 +/- 1.0 to 4.0 +/- 0.6 mV/sec (p less than 0.001). Preincubation of normal whole blood with fat emulsion in vitro did not adversely affect chemiluminescence (11 studies), nor did incubation of normal neutrophils with patient postinfusion plasma (10 studies). We conclude that fat emulsion infusion acutely suppresses neutrophil chemiluminescence. The suppression is not a direct effect of the fat emulsion per se and is not due to inhibitory substances in the plasma following infusion.     

Evaluation of peripheral blood neutrophil leucocytes in lead-exposed workers

Background: Inorganic lead can interfere with humoral and especially cell-mediated immunity even at frequently occurring (<50 μg/dl) blood lead (Pb-B) levels. Occupational exposure to lead causes a primary impairment of the chemotactic and phagocytic activities of neutrophil leucocytes. Objective: To verify whether, after taking into account the main confounding factors, occupational lead exposure is shown to induce changes in the number of blood neutrophil leucocytes, and to assess a possible dose-response relationship between Pb-B and the circulating neutrophil count in exposed workers. Subjects and methods: The study included 68 male lead-exposed (E) workers and 59 male workers in a food plant, recruited as controls (NE). A standardized questionnaire probing work, social, familial and personal medical history was administered to all the subjects. Blood and urine samples were collected to determine the dose and effect biological indices of lead and the total white blood cell and neutrophil counts. Results: Pb-B levels were significantly higher in E (geometric mean (GM): 20.5 μg/dl; 3.2–120 μg/dl) than in NE workers (GM: 3.5 μg/dl; 1–11 μg/dl). The mean absolute neutrophil count (ANC) was significantly higher in E workers with respect to NE workers. ANC correlated significantly with the biological lead dose and effect indices. Moreover, there was a dose-dependent increase of ANC with increasing Pb-B levels. The linear relationship between ANC and Pb-B was confirmed even after correction for age, body mass index and smoking habit. We also found an interaction between Pb-B level and smoking habit in increasing the number of blood neutrophils in lead-exposed workers. Conclusion: Our study is the first to describe a dose-dependent effect of lead on ANC in exposed workers. Our results underline the importance of promoting a further reduction of occupational lead exposure levels, adopting adequate individual protection means, as well as conducting medical campaigns against smoking, at the workplace.     

Chronic effect of training on neutrophil functions in humans

We performed this study to evaluate the chronic effect of training on neutrophil functions in humans. Twenty-six university students (14 males and 12 females) with a mean age of 19.1±0.8 years were divided into the athlete group and the control group based on answers given to a written questionnaire. The capacity of circulating neutrophils to ingest bacteria (phagocytosis) and to produce superoxide (nitroblue tetrazolium reduction) was measured under resting conditions in the absence of training activities. In addition, we measured hematological and serum biochemical parameters.The present analyses revealed that the frequency of subjective symptoms concerning susceptibility to infections (sum of males and females) was higher in the athlete group than in the control group (p<0.05). In the investigation of males, hemoglobin and serum protein levels of the athlete group were lower than that of the control group (p<0.01, p<0.05, respectively). Although total and differential leukocyte counts, and phagocytic activity of neutrophils were not significantly different between the groups, superoxide productivity of neutrophils in the male athlete group was higher than that in the control group (p<0.05).Since significant differences were observed in hemoglobin and serum protein levels in the male athletes, the training is considered to have been intense. An increase was noted in the neutrophil superoxide production along with these changes, but no significant difference was observed in the phagocytic activity. Therefore, there is the possibility of superoxide overproduction, which may lead to tissue damage.