Investigation of K+ channel expression in human peripheral lymphocytes of healthy donors by means of flow cytometry

Evaluation of different types of K+ channel expression was performed in resting and PHA (phytohemagglutinine)-activated human peripheral lymphocytes (HPL) of healthy donors by means of flow cytometry. In resting peripheral lymphocytes, the application of kaliotoxin (a selective blocker for voltage-dependent K+ (K(V)) channels), K(V) resulted in pronounced depolarization of lymphocyte membrane potential, with further promotion in the presence of thapsigargin (compound discharging Ca(i) from endoplasmic reticulum). In activated HPL, the expression of various types of K+ channels was estimated utilizing cell-cycle analysis data. In contrast to the resting cells, kaliotoxin-induced depolarization of membrane potential in PHA-activated lymphocytes of the G0/G1 phase was not enhanced by thapsigargin and in PHA-activated lymphocytes of the S and G2/M phases we were able to observe repolarization of membrane potential after kaliotoxin-induced depolarization of membrane potential. Substitution of kaliotoxin for charybdotoxin (a non-selective drug blocking both K(V) and K(Ca) channels) abrogated the above effects in PHA-activated lymphocytes. Thus, K(V) channels are active in both resting and activated HPLs and K(Ca) channel expression occurs with cell-cycle progress on PHA-induced activation of peripheral lymphocytes.     

Phenotypic analysis of human peripheral blood lymphocytes by automatic sampling flow cytometry after stimulation with mitogens or allogeneic cells

Summary Human peripheral blood lymphocyte (PBL) phenotypes have been analyzed before and after stimulation with phytohemagglutinin (PHA), concanavalin A (ConA) and pokeweed mitogen (PWM) for 3 days and in mixed lymphocyte culture (MLC) for 7 days. PBL labeled with each of 10 fluorescent monoclonal antibodies were automatically sampled for flow cytometry from 96-well microtiter plates using a microsample delivery system. The reference phenotypic ranges were determined in fresh cells and control cultures. PHA was mostly mitogenic for T PBL bearing the CD3, CD5, CD7, CD8 and CD25 differentiation clusters, and a low density of CD1 and CD4 had a small effect on human natural killer cells (HNK) and also did not stimulate B (CD19) and HLA-DR⁺ PBL. There was an incomplete phenotypic overlapping between PHA- and ConA-stimulated cultures, ConA being more mitogenic for CD4 and less mitogenic for CD8 PBL. The mitogenic effect of PWM was evident on CD3, CD5, CD7, CD4, CD25 and CD8, but not on HNK, HLA-DR and CD19 B PBL, which presumably had already differentiated into antibody-secreting cells. After MLC stimulation all T, B and HNK PBL subsets tested were increased, but the cells bearing CD1, CD4, CD5, CD7, CD25, HNK, CD19 and HLA-DR had the greatest proliferation with respect to the unmixed control PBL. The present approach to the phenotyping of PBL subsets could offer more complete and accurate data for monitoring and follow-up of patients in transplantation and immunopathology hospital wards. This work was supported in part by grants from theConsiglio Nazionale delle Ricerche (CNR), Roma, Italy,Progetto Finalizzato ‘Oncologia’, theAssociazione Italiana per la Ricerca sul Cancro (AIRC), and theAssociazione per le Ricerche Biomediche, Verona, Italy.     

多器官功能障碍评分系统:3个评分标准预测多器官功能障碍综合征结局关联性和准确性的比较与评估

目的研究高原急性呼吸窘迫综合征(HARDS)/多器官功能障碍综合征(MODS)各项诊断指标参数的变化特点,比较3个MODS评分标准预测结局的准确性。方法统一按通用的MODS诊断标准将540例ARDS/MODS患者按海拔高度分为平原对照组(CG,<430m,n=113)、中度高原1组(H1G,1517m,n=314)、中度高原2组(H2G,2261～2400m,n=78)和高原组(HG,2808～3400m,n=35)。4组分别用平原地区ARDS/MODS评分诊断标准(庐山会议评分标准和Marshall评分标准)以及兰州修订的HARDS/MODS评分标准(兰州标准),建立3个标准的数据统计模型,分别绘制受试者运行特征性曲线(ROC曲线),计算约登指数(Yoden)和最佳界值,验证3个标准在不同海拔高度预测ARDS/MODS结局的准确性;用向前逐步回归模式对影响MODS结局的多因素进行分析。结果用庐山、Marshall和兰州标准检验平原和高原不同海拔高度MODS总分的ROC曲线下面积,预测结局的敏感度、特异度及其最佳界值,结果显示,随海拔梯度上升,兰州标准明显优于庐山和Marshall标准,多元Logistic回归分析也以兰州标准的影响因素最大。结论1通用的ARDS/MODS诊断标准中某些参数界值可能不适合中度高原以上地区,建立HARDS/MODS标准是必要的,兰州标准随海拔梯度升高有进一步提高预测准确性的趋势。2海拔高度大于1500m以上地区     

Vascular endothelial growth factor receptor and coreceptor expression in human acute respiratory distress syndrome

Background

Acute respiratory distress syndrome (ARDS) is characterized by the development of noncardiogenic pulmonary edema, which has been related to the bioactivity of vascular endothelial growth factor (VEGF). Vascular endothelial growth factor receptors and coreceptors regulate this bioactivity. We hypothesized VEGF receptors 1 and 2 (VEGFR1, VEGFR2) and coreceptor neuropilin-1 (NRP-1) would be expressed in human lung tissue with a significant change in expression in ARDS lung.

Methods

Archival “normal” (no lung pathology and non-ARDS), “early” (within 48 hours), and “later” (after day 7) ARDS lung-tissue sections (n = 5) were immunostained for VEGFR1, VEGFR2, and NRP-1 from human subjects (n = 4). Staining was assessed densitometrically using Histometrix software.

Results

VEGFR1, VEGFR2, and NRP-1 were expressed on both sides of the alveolar-capillary membrane in both normal and ARDS human lung tissue. In later ARDS, there was a significant up-regulation of VEGFR1 and VEGFR2 versus normal and early ARDS (P < .0001). Neuropilin-1 was down-regulated in early ARDS versus normal lung (P < .05), with normalization in later ARDS (P < .001).

Conclusion

Differential temporal VEGFR1, VEGFR2, and NRP-1 up-regulation occurs in human ARDS, providing evidence of further functional regulation of VEGF bioactivity via VEGFR2 consistent with a protective role for VEGF in lung injury recovery. The mechanisms behind these observations remain to be clarified.     

控制性肺膨胀对急性呼吸窘迫综合征家兔肺外器官炎症反应的影响

目的 研究控制性肺膨胀(SI)对急性呼吸窘迫综合征(ARDS)家免肺外器官炎症反应的影响。方法 生理盐水肺泡灌洗法复制ARDS家兔模型。实验动物随机分为对照组、ARDS组、小潮气量(VT) 最佳呼气末正压组(LVBP组)和SI组。机械通气4 h后,逆转录-聚合酶链反应(RT-PCR)检测肝、肠组织肿瘤坏死因子-α(TNF-α)和自细胞介素-10(IL-10)mRNA表达;酶联免疫吸附法检测肝、肠组织TNF-α和IL-10浓度,同时测定髓过氧化物酶(MPO)和丙二醛(MDA)含量。结果 SI组肝、肠组织TNF-α mRNA表达明显低于ARDS组和LVBP组(P均<0.05);肝、肠组织IL-10 mRNA表达均显著低于ARDS组和LVBP组(P均<0.05)。各组间肝、肠组织TNF-α、IL-10、MPO和MDA含量比较无统计学差异。结论 SI可抑制ARDS家免肺外器官细胞因子mRNA的表达,下调肺外器官炎症反应。     

肺保护性通气对急性呼吸窘迫综合征家兔肺外器官炎症反应的影响

目的　研究肺保护性通气对急性呼吸窘迫综合征 (ARDS)肺外器官炎症反应的影响。方法　用生理盐水肺泡灌洗法复制ARDS家兔模型 ,分为 6组 (1)正常对照组 ;(2 )ARDS模型组 (ARDS) ;(3)小潮气量 (VT) +最佳呼气末正压 (PEEP)组 (LVBP) ;(4)常规VT+最佳PEEP组 (NVBP) ;(5 )小VT+高PEEP组 (LVHP) ;(6 )高VT+零PEEP组 (HVZP)。后 4组机械通气 4h后 ,用逆转录PCR法 (RT PCR)检测动物肝肠组织肿瘤坏死因子 (TNF)α和白介素 (IL) 10mRNA表达 ,酶联免疫吸附法 (ELISA)测定肝肠组织TNF α及IL 10浓度。结果　LVBP组肝组织TNF α和IL 10mRNA表达分别为 (42± 9)和 (2 5±10 ) ,与ARDS模型组相比较 ,差异无显著性 ,分别为 [(37± 7)和 (2 5± 4 ) ],但显著低于NVBP [分别(5 6± 7)和 (36± 8) ]、LVHP [(5 3± 10 )和 (35± 5 ) ]和HVZP组 [(70± 10 )和 (46± 5 ) ]。LVHP组TNF α和IL 10mRNA表达明显高于LVBP组 ,但明显低于HVZP组。LVBP组肝组织TNF α和IL 10浓度与ARDS组无明显差异 ,但显著低于NVBP、LVHP和HVZP组。HVZP组肝组织TNF α和IL 10浓度最高 ,不但高于LVBP组 ,也高于LVHP组。LVBP组的肠组织的TNF αmRNA表达为 (38± 11) ,与ARDS组比较无显著差异 ,但显著低于NVBP (5 1± 9)、LVHP (5 0± 11)和HVZP组     

A Critical Appraisal of the Effects of Anesthetics on Immune-system Modulation in Critically Ill Patients With COVID-19

PurposeThe aim of the present article was to briefly summarize current knowledge about the immunomodulatory effects of general anesthetics and the possible clinical effects of this immunomodulation in patients with COVID-19.MethodsThe PubMed, Scopus, and Google Scholar databases were comprehensively searched for relevant studies.FindingsThe novel coronavirus causes a wide spectrum of clinical manifestations, with a large absolute number of patients experiencing severe pneumonia and rapid progression to acute respiratory distress syndrome and multiple organ failure. In these patients, the equilibrium of the inflammatory response is a major determinant of survival. The impact of anesthetics on immune-system modulation may vary and includes both pro-inflammatory and anti-inflammatory effects.ImplicationsInhibition of the development of severe inflammation and/or the enhancement of inflammation resolution by anesthetics may limit organ damage and improve outcomes in patients with COVID-19.     

体外膜肺对急性呼吸窘迫综合征幼猪肺生理和病理的影响

目的研究体外膜肺（ECMO）对健康及油酸致急性呼吸窘迫综合征幼猪呼吸力学、氧合、肺部炎症反应及病理形态的影响。方法21只健康幼猪随机分为4组：CON组（n=5）;ECMO组（n=6）：机械通气＋ECMO24h后观察6h;ARDS组（n=5）;AEC组（n=5）：成模后以机械通气＋ECMO24h,观察6h。在基础状态（Base）、成模时（Oh）、以后每小时监测血气、血流动力学和呼吸力学,检测肺湿干重比（W／D）,肺泡灌洗液（BALF）中总蛋白（TP）、总磷脂（TPL）、饱和磷脂（DSPC）、最小表面张力（γmin）,肺组织TNF-α、IL-1β、IL-6、IL-8、IL-10 mRNA表达,及肺病理学改变。计数资料比较用方差分析,等级资料多组间差别用Ktudksl-Wallis检验。P〈0．05视为差异具有统计学意义。结果（1）CON组和ECMO组全部存活,AEC组1只死亡,ARDS组全部于成模后24h内死亡。（2）ARDS组和AEC组成模后0I和Aa-DO2显著升高[OI：（7．5±1．0）vs（10．5±3．8）;AaDO2：（556．0±24．3）vs（558．0±18．3）],随后ARDS组OI和Aa-DO2进一步增高,而AEC组在ECMO启动后两者下降[8hOI：（18．2±3．6）vs（7．6±9．3）;Aa-DO2：（574．0±12．8）vs（329．0±279．0）]。（3）与CON组比较,ARDS组和AEC组TPL下降,γmin升高,差异具有统计学意义（P〈0．01）。与CON组和ECMO组比较,ARDS组IL-6和IL-8mRNA表达显著增高（P〈0．05）。（4）ECMO组有轻度肺病理损伤。与ARDS组相比,AEC组各种病变均减轻,肺泡扩张度改善。结论ECMO转流24h能改善ARDS幼猪氧合和气体交换,减轻肺部病理损伤,提高生存率;激活健康幼猪肺部炎症反应,引起轻度肺病理损伤。     

用流式细胞术研究血液病细胞死亡生物学

Darzynkiewicz等提出的细胞死亡生物学,意在研究细胞死亡前、中、后的形态、生化和分子生物学等的全过程以及细胞死后邻近组织的反应。经过半个多世纪对细胞增殖动力学的研究后,人们终于认识到细胞死亡与增殖同等重要。两者的不平衡是多种疾病的根源,因而细胞死亡的研究成为近年来生命科学的研究热点。细胞凋亡在骨髓增生异常综合征,慢性淋巴细胞白血病,慢性粒细胞白血病,以及急性白血病发生发展中的作用已获得初步证实。深入研究其机理有利于发展新的靶向治疗。为了准确判断细胞死亡类型,可靠的实验技术至关重要。流式细胞术(FCM)能多参数、快速、定性、定量测定群体细胞中单个细胞的生物学过程,例如用光散射可测定细胞形态,用膜联蛋白(Annexin V),碘化丙啶(PI)和吖啶橙(AO)等,可测定早期细胞膜结构与通透功能。若丹明123(rhodamine 123)是线粒体特异标志。DNA经内切酶降解后在FCM-DNA图中出现亚G_1峰。用各种抗基因蛋白单克隆抗体如Bcl-2,Bcl-XL,Bax,ICE,c-myc,Fas/FasL等可测知单个细胞中与细胞凋亡相关基因蛋白表达及其间关系。Tunnel法与PI双标志法是判断凋亡细胞发生在哪个周期时相的最可靠方法。本文简要介绍各种方法的原理与质量控制以及如何正确判断实验结果。     

急性肾损伤时水通道蛋白2及钠钾泵的表达

目的 探讨水通道蛋白2的表达及钠-钾-三磷酸腺苷酶在多器官功能障碍肾损伤发病机制中的作用,研究大鼠多器官功能障碍肾损伤与水通道蛋白2及钠-钾-三磷酸腺苷酶的表达.方法 选取健康大鼠72只,随机(随机数字法)分为对照组24只,脂多糖组48只,采用大鼠腹腔注射脂多糖5 mg/kg造成内毒素致多器官功能障碍动物模型,对照组仅作假手术处理.造模成功后6h、12h、24 h、2d、3d、5d分别处死动物,留取血液、尿量进行生化检测,并应用免疫组化法和RT-PCR技术检测肾组织内水通道蛋白2 mRNA及蛋白的表达水平.应用试剂盒测定钠-钾-三磷酸腺苷酶的含量及活性.结果 造模成功后致伤组大鼠尿量迅速减少,48 h后尿量增多.尿素氮、肌酐逐渐增高,48 h达高峰,此后逐渐下降.水通道蛋白2 mRNA及蛋白表达迅速减少,48 h降至最低,此后逐渐增多.钠-钾-三磷酸腺苷酶含量差异无统计学意义,但其活性在造模成功后明显降低,此后逐渐增高,但仍低于对照组.结论 大鼠多器官功能障碍综合征肾损伤模型中,水通道蛋白2是肾脏重吸收功能恢复的结构基础,钠-钾-三磷酸腺苷酶则直接参与或间接反映肾脏的能量代谢状态,只有在水通道蛋白2及能量代谢恢复后,大鼠的肾脏功能才能逐渐好转.     

全身炎症反应综合征-急性肺损伤大鼠肺组织IL-4 mRNA表达及AP-1活性变化的研究

目的 复制脂多糖（LPS）致伤的全身炎症反应综合征（SIRS）-急性肺损伤（ALI）的大鼠模型。观察肺组织白介素-4（IL-4）mRNA表达量和激活蛋白-1（AP-1）活性的变化，探讨SIRS-ALI中抗炎机制及其调控的意义。方法 经Wistar大鼠尾静脉注射递增剂量LPS，复制SIRS-ALI大鼠模型；逆转录PCR法（RT-PCR）检测肺组织IL-4mRNA表达量；凝胶迁移率分析法（EMSAs)检测肺组织AP-1活性。结果 （1）LPS可以介导大鼠发生SIRS-ALI；（2）LPS≥6mg/kg可致SIRS-ALI失控，形成ARDS；（3）LPS可诱导肺组织IL-4mRNA表达量和AP-1活性升高；（4）LPS≥6mg/kg时，肺组织IL-4mRNA表达量和AP-1活性的升高幅度最大。结论 （1）LPS≥6mg/kg是大鼠SIRS-ALI发生失控的临界剂量；（2）SIRS-ALI失控伴有IL-4基因转录水平明显上调，可能与上游调控因子AP-1活性异常增强有关；（3）抗炎机制增强在SIRS-ALI发生，发展过程中发挥了重要作用。     

肺内源性与外源性急性呼吸窘迫综合征模型猪的肺复张性比较

目的应用分层电阻抗成像技术（EIT）比较肺内源性急性呼吸窘迫综合征（ARDSp）和肺外源性急性呼吸窘迫综合征（ARDSexp）模型猪的肺复张性。 方法健康巴马猪16只,随机分为ARDSp组和ARDSexp组,每组各8只。采用盐酸气管吸入的方法复制ARDSp模型,油酸静脉滴注的方法复制ARDSexp模型。模型复制成功后,采用肺保护性通气策略进行机械通气并进行一次肺复张,之后保持其他呼吸机参数不变,分别在高低呼气末正压（PEEP）25 cmH₂O和5 cmH₂O水平下通气1 h,通气结束后进行吸气末和呼气末暂停,记录高低PEEP水平间的复张容积、每个PEEP水平下的氧合情况及血流动力学参数。 结果ARDSexp组和ARDSp组模型的复张容积为分别为（1572.8±314.9）ml和（1089.9±224.2）ml,差异有统计学意义（P=0.003）。升高PEEP能够改善ARDSexp的氧合指数,由（214.3±128.6）mmHg提高到（447.5±96.0）mmHg,差异有统计学意义（P=0.004）,而ARDSp高低PEEP水平间氧合指数变化不显著,差异无统计学意义［（173.6±112.0）mmHg vs （178.4±128.2）mmHg,P=0.943］。 结论ARDSexp和ARDSp模型猪的复张性不同,相比ARDSp,ARDSexp的复张性更好。高PEEP水平可能能够改善ARDSexp的氧合,而对ARDSp的效应相对不显著。     

动态通气参数对急性呼吸窘迫综合征犬肺外炎症反应和肺外脏器功能的影响

目的 观察机械通气动态通气参数对急性呼吸窘迫综合征(ARDS)犬肺外炎症介质水平及肺外脏器功能的影响.方法 36条健康杂种犬,按照随机数字表法分为正常对照组(N组)、ARDS模型组(M组)及机械通气A-D组6组.采用气管内盐酸吸入法建立ARDS模型,按下述方案行机械通气.A组(LVLYHR):小潮气量、低吸气流速、高通气频率;B组(HVHFHR):大潮气量、高吸气流速、高通气频率;C组(HVHFLR):大潮气量、高吸气流速、低通气频率;D组(HVIYLR):大潮气量、低吸气流速、低通气频率.分组机械通气后4 h后处死动物,留取血清用放射性免疫吸附法行IL-8和TNF-α检测;留取肝肾组织行病理学检查.数据处理运用方差分析,以P<0.05为差异具有统计学意义.结果 B组、C组血中IL-8和TNF-α含量显著高于M组、A组、D组(P<0.05),A组、D组和M组差异无统计学意义(P>0.05).N组与其他相比差异具有统计学意义(P<0.05).B组肝肾组织病理学改变最为严重,C组较B组稍有减轻.A组和D组的病理学改变较B、C组明显减轻,A组和M组改变接近.结论 大潮气量、高吸气流速、高通气频率机械通气可以使血清炎症介质水平升高,加重肺外器官的炎症反应;降低通气频率及吸气流速,对防治多器官功能衰竭有重要意义.     

白细胞介素-10对急性肺损伤炎症/抗炎介质表达的影响

目的探讨白细胞介素-10(IL—10)对急性肺损伤(ALI)大鼠炎症介质／抗炎介质表达的影响。方法向气道内滴注内毒素(LPS，10mg／kg)建立大鼠ALI模型。54只雄性SD大鼠随机分为对照组、LPS损伤组、LPS加IL-10组，每组18只，各组又分为2、6和24h3个亚组，每个亚组各6只。按各时间点观察大鼠动脉血氧分压(PaO2)、支气管肺泡灌洗液(BALF)中细胞总数及分类计数、肺系数、BALF总蛋白水平及肺病理，同时用逆转录-聚合酶链反应(RT—PCR)方法检测肺组织中炎症介质／抗炎介质的表达。结果①LPS损伤组大鼠PaO2呈进行性降低；肺系数、BALF总蛋白水平及BALF中细胞总数均明显增加，分类以中性粒细胞为主；肺病理示肺内中性粒细胞大量浸润，伴出血、透明膜形成。LPS加IL-10组的各项指标均较LPS损伤组减轻。②LPS损伤组肺组织肿瘤坏死因子-α(TNF—α)mRNA表达于2h达高峰，随后迅速下降；白细胞介素-1β(IL—1β)mRNA表达于2h显著升高，6h达高峰，随后迅速下降；IL-1受体拮抗剂(IL—1ra)mRNA表达6h开始升高，且为峰值。24h仍高于对照组。LPS加IL-10组肺组织TNF—αmRNA、IL-1βmRNA表达受抑，而IL—1ra mRNA表达不受影响。结论①ALI早期TNF—αmRNA、IL-1βmRNA表达明显增加，而IL—1ra mRNA表达滞后，提示在无外来干预情况下，ALI早期存在炎症介质／抗炎介质的失衡。②IL-10可明显抑制炎症介质表达，不影响抗炎介质表达，有利于重建炎症介质／抗炎介质平衡，减轻LPS所致ALI。     

Activation of Ca2+-dependent K+ channels in human B lymphocytes by anti-immunoglobulin.

Many mammalian cell types exhibit Ca2+-dependent K+ channels, and activation of these channels by increasing intracellular calcium generally leads to a hyperpolarization of the plasma membrane. Their presence in B lymphocytes is as yet uncertain. Crosslinking Ig on the surface of B lymphocytes is known to increase the level of free cytoplasmic calcium ([Ca2+]i). However, rather than hyperpolarization, a depolarization has been reported to occur after treatment of B lymphocytes with anti-Ig. To determine if Ca2+-dependent K+ channels are present in B lymphocytes, and to examine the relationship between intracellular free calcium and membrane potential, we monitored [Ca2+]i by means of indo-1 and transmembrane potential using bis(1,3-diethylthiobarbituric)trimethine oxonol in human tonsillar B cells activated by anti-IgM. Treatment with anti-IgM induced a biphasic increase in [Ca2+]i and a simultaneous hyperpolarization. A similar hyperpolarization was induced by ionomycin, a Ca2+ ionophore. Delaying the development of the [Ca2+]i response by increasing the cytoplasmic Ca2+-buffering power delayed the hyperpolarization. Conversely, eliminating the sustained phase of the [Ca2+]i response by omission of external Ca2+ abolished the prolonged hyperpolarization. In fact, a sizable Na+-dependent depolarization was unmasked. This study demonstrates that in human B lymphocytes, Ca2+-dependent K+ channels can be activated by crosslinking of surface IgM. Moreover, it is likely that, by analogy with voltage-sensitive Ca2+ channels, Na+ can permeate through these ligand-gated Ca2+ "channels" in the absence of extracellular Ca2+.     

传染性非典型肺炎患者外周血CD4+T细胞 CD38的表达及意义

目的 探讨传染性非典型肺炎（急性重症呼吸综合征，SARS）患者外周血CD4＋T细胞CD38的表达的变化及其与病程和病情严重程度的关系。方法 用双色流式细胞术测定157例SARS临床诊断病例、37例同期发生的肺炎支原体感染患者及38名健康成人外周血淋巴细胞中CD4＋T细胞的百分率（CD4％）、CD4＋T细胞CD38阳性率（CD38／CD4％）及CD38平均荧光强度（CD38MF）进行检测。结果 SARS病例CD4％与健康对照组相比均无统计学意义，但其CD38／CD4％及CD4＋T细胞CD38 MF显著低于健康对照组（P〈0．05，P〈0．05）；SARS重症患者CD38／CD4％及CD38MF显著低于轻症患，有统计学意义（P〈0．01，P〈0．001）；SARS轻症患者CD4％、CD38／CD4％及CD38MF与健康对照组相比无统计学意义；SARS临床诊断病例CD4％显著高于肺炎支原体感染者，有统计学意义（P〈0．01），而CD38／CD4％及CD38MF显著低于肺炎支原体感染，有统计学意义（P〈0．01，P〈0．01）。结论 SARS患者CD4＋T细胞CD38的表达降低，降低的水平与SARS病情严重程度相关。CD4＋T细胞CD38的表达在SARS诊断病例及肺炎支原体感染中的变化存在统计学意义，CD4＋T细胞CD38的表达的检测有助于SAILS与肺炎支原体感染的鉴别。     

ARDS患者肺组织Bax和Bcl-2表达的变化及意义

目的　探讨Bax和Bcl- 2表达的变化在ARDS发病中的作用 ,试图为进一步阐明ARDS的发病机制和在细胞凋亡水平防治本病提供理论依据。方法　ARDS组 9例 ,对照组 5例 ,采用TUNEL法以及免疫组化技术观察ARDS患者肺组织细胞凋亡及Bax和Bcl- 2表达的变化。结果　ARDS急性期患者肺组织细胞凋亡率较对照组明显增加 ,且主要表现为肺泡上皮和肺血管内皮细胞凋亡增加 ;Bcl- 2表达ARDS组和对照组无显著差异 ,Bax在ARDS急性期患者肺组织表达明显上调 ,并且Bax表达的增加与肺组织细胞凋亡增加呈平行关系。结论　肺泡上皮和肺血管内皮细胞凋亡改变和Bax/Bcl- 2表达的变化可能参与临床ARDS急性期的发病。     

Sequential production of leukaemia inhibitory factor by blood cell culture in patients with ARDS

Objective: Leukaemia inhibitory factor (LIF) is a polyfunctional cytokine integrated in cytokine networks and its concentration has been shown to be elevated in bronchoalveolar lavage fluid of patients with the acute respiratory distress syndrome (ARDS). The aim of our study was to evaluate the production of LIF by culturing blood cells from patients with ARDS. Patients: 8 patients with ARDS, 8 patients with pneumonia and 5 healthy subjects. Measurements and results: The blood samples were taken on day 1 after onset of ARDS. LIF was measured, in the cell-free supernatant, with an enzyme-linked immunosorbent assay after 24 h, 48 h and 72 h of blood cell culture. LIF was detectable in some patients in the ARDS group: at i) at 24 h and 48 h: in 2 patients ii) at 72 h in 4/5 patients (140 ± 231 pg/ml). Only in the 4 patients in whom LIF was measured at 72 h was ARDS associated with the multiple organ dysfunction syndrome. Furthermore, among the 5 patients with ARDS who subsequently died, 4 had a detectable LIF. Conclusions: We have observed that LIF was produced only in ARDS, but not in all patients. The production of LIF seems to be a good indicator of the severity of ARDS. These preliminary results must be confirmed by a larger study. Received: 31 July 1997 Accepted: 13 January 1998     

Outcome value of Clara cell protein in serum of patients with acute respiratory distress syndrome

Objective Injury to the alveolocapillary barrier characterizes ALI/ARDS; therefore determining levels of lung epithelium-specific small proteins in serum may help predict clinical outcomes. We examined whether serum Clara cell protein (CC-16) concentration is correlated with the outcome, mechanical ventilation duration, and incidence of nonpulmonary organ failure.Design Prospective multicenter observational study conducted by the Quebec Critical Care Network.Measurements Seventy-eight adult ARDS patients requiring mechanical ventilation were enrolled and 28-day mortality was the primary outcome. Ventilatory parameters were computed and blood was sampled daily. Clinical information collected included cause of death, duration of mechanical ventilation, number of ventilator-free days, and organ failures.Results Median serum levels of CC-16 were significantly higher in nonsurvivors than survivors on days 0–2 (19.93 μg/l, IQR 11.8–44.32, vs. 8.9, 5.66–26.38) and sustained up to day 14. CC-16 levels were correlated positively with the number of failing organs (ρ = 0.3623) and requirement for prolonged mechanical ventilation. Predictors of patient mortality included age, arterial carbon dioxide partial pressure, CC-16, and APACHE II score (odds ratios 1.35, 1.52, 1.37, 1.159, respectively).Conclusions Higher initial CC-16 serum level is associated with increased risk of death, fewer ventilator-free days, and increased frequency of nonpulmonary multiple organ failure. CC-16 is a valuable biomarker of ARDS that may help predict outcome among ARDS patients with high-risk mortality.Financial support was provided by the Réseau en Santé Respiratoire du FRSQ (O.L.). O.L. and G.B. are research scholars, and E.R. is a national scholar of the FRSQ.This article is discussed in the editorial available at:     

血栓素B2及血浆内皮素-1在急性呼吸窘迫综合征中的表达及临床意义

目的研究血栓素B2(TXB2)及血浆内皮素-1(ET-1)在急性呼吸窘迫综合征(ARDS)患者中表达差异。方法选自我院收入的ARDS患者共45例,另选30例健康人群作为健康对照,ARDS组患者分别在确诊后1d、4d和7d清晨三个不同时点各抽取空腹静脉血5ml,健康对照组同样于清晨空腹抽静脉血5ml,采用放射免疫法测定血浆中的TXB2及血浆ET-1,所测结果两组进行比较。结果确诊后1d、4d和7d的血浆ET-1表达水平显著高于健康对照组(P<0.05)。ARDS组患者的TXB2于确诊后1d、4d和7d表达水平显著高于健康对照组(P<0.05)。确诊后7dTXB2表达水平达到最高值,显著高于1d和4d两个时点(P<0.05)。结论 ARDS患者血浆中TXB2及血浆ET-1水平明显增高,TXB2及血浆ET-1可能参与了ARDS发生的病理生理过程,其含量变化与疾病的发生发展可能存在关系,为诊断ARDS提供了一个有价值的指标。