人血清及中性粒细胞的乳铁蛋白含量与中性粒细胞吞噬…

用ＥＬＩＳＡ法测定了人血清中及中性粒细胞（ＰＭＮ）中乳铁蛋白（ＬＦ）含量，并与ＰＭＮ的吞噬功能比较，证实血清中ＬＦ水平与ＰＭＮ吞噬功能的不相关，ＰＭＮ中ＬＦ含量与其吞噬，杀菌功能有密切的正相关关系。     

铁与中性粒细胞的杀菌功能

铁与中性粒细胞的杀菌功能王景文张之南李蓉生中性粒细胞（ＰＭＮ）在机体抗感染的防御反应过程中起重要作用。ＰＭＮ的杀菌功能依靠几个不同机制，大体说：一种是依靠ＰＭＮ产生活性氧的赖氧杀菌机制。ＰＭＮ吞噬异物后，发生急剧的反应称呼吸爆发，表现为耗氧量快速增加...     

正常及PNH患者中性粒细胞氧化杀菌功能与其表面FcγRⅢb(CD_(16b))关系的探讨

正常及ＰＮＨ患者中性粒细胞氧化杀菌功能与其表面ＦｃγＲⅢｂ（ＣＤ１６ｂ）关系的探讨傅新容崔毅许彩民潘华珍张之南中性粒细胞（ＰＭＮ）是机体防御系统的主要组成部分，能吞噬和杀灭多种致病原。致病原与血清中的特异性抗体（ＩｇＧ）结合后，通过ＩｇＧ的Ｆｃ部分与...     

脓毒血症患者中性粒细胞变形性检测结果分析

为了解中性粒细胞（ＰＭＮ）在脓毒血症中的作用，采用ＰＭＮ核孔膜滤过的方法，对１３例脓毒血症患者与５０例正常人的ＰＭＮ变形能力进行了测定，以ＰＭＮ滤过指数（ＦＩ）表示。结果：脓毒血症组患者ＰＭＮ的ＦＩ明显高于正常对照组（Ｐ＜０．０１）；其中死亡患者ＰＭＮ的ＦＩ则明显高于存活组（Ｐ＜０．０５）；存活组与死亡组ＦＩ的临界值为１３．３８，当ＦＩ＜１３．３８时，患者全部存活；当ＦＩ≥１３．３８时，病死率明显上升（Ｐ＜０．０１）。说明脓毒血症患者的ＰＭＮ变形能力明显下降，并与病情的转归有密切关系，当下降到一定程度时，其病死率明显升高。因此，ＰＭＮ变形性的检测可能成为预测脓毒血症患者预后的重要指标之一；同时又可为临床选择治疗方法提供参考依据     

围急性呼吸窘迫综合征期中性粒细胞化学发光的改变及其意义

目的：了解急性肺损伤形成和发展过程中中性粒细胞的功能状况。方法：采用液闪单光子辐射法检测全身炎性反应综合征（ＳＩＲＳ）急性呼吸窘迫综合征（ＡＲＤＳ）多脏器功能失常综合征（ＭＯＤＳ）患者中性粒细胞（ＰＭＮ）的化学发光。结果：处于ＳＩＲＳ期患者ＰＭＮ的基础发光和最大吞噬发光均有所增强；进入ＡＲＤＳ期的患者ＰＭＮ基础发光仍处高水平，但最大吞噬发光已有所下降；进一步发展至ＭＯＤＳ期的患者基础发光无明显下降，而最大吞噬发光已下降至明显低于静息ＰＭＮ水平。结论：机体全身炎性反应早期ＰＭＮ的激活水平增高，吞噬反应能力增强；而随病情进展及恶化，在ＰＭＮ的激活维持高水平的同时，其吞噬反应能力则逐渐下降乃至衰竭。     

脓毒血症患者中性粒细胞为形性检测结果分析

为了解中性粒细胞在脓毒血症中的作用，采用ＰＭＮ核孔膜滤过的方法，对１３例脓毒血症患者与５０全正常人的ＰＭＮ变形能力进行了测定，以ＰＭＮ滤守指数表示。结果；脓毒血症组患者ＰＭ怕ＦＩ明显高于正常对照组；其中死亡患者ＰＭＮ的ＦＩ则明显高于存活组；存活组与死亡组ＦＩ的临界值为１３．３８，当ＦＩ〈１３．３８时，患者全部存活，当ＦＩ≥１３．３８％时，病死率明显上升。     

系统性红斑狼疮中抗中性粒细胞胞浆抗体的检测

应用间接免疫荧光技术（ＩＩＦ），对系统性红斑狼疮（ＳＬＥ）患者进行了抗中性粒细胞胞浆抗体（ＡＮＣＡ）的检测，并以３５例正常体检者为对照。结果表明在１３１例ＳＬＥ患者中，ＡＮＣＡ阳性为２２例，阳性率为１６．５％，其中Ｃ－ＡＮＣＡ阳性１６例占１２．２％，Ｐ－ＡＮＣＡ阳性６例占４．６％，对ＳＬＥ患者进行ＡＮＣＡ的检测有助于疾病的诊断。     

肺原性心脏病急性加重期一氧化氮及肿瘤坏死因子的研究

目的：探讨一氧化氮（ＮＯ）和肿瘤坏死因子（ＴＮＦ）在肺原性心脏病（肺心病）发病中的作用和意义。方法：分别用镀铜镉还原法和放射免疫分析法测定３０例肺心病急性期患者治疗前后血清ＮＯ及ＴＮＦ含量。结果：肺心病急性期患者血清ＮＯ和ＴＮＦ水平均较缓解期（治疗后）及正常人组显著增高（ＮＯ－２／ＮＯ－３分别为：１３８．５±５６．７μｍｏｌ／Ｌ与７７．４±４８．６μｍｏｌ／Ｌ及３４．６±１７．８μｍｏｌ／Ｌ之比）；合并呼衰组血清ＮＯ和ＴＮＦ水平较非呼衰组均明显增高（ＮＯ－２／ＮＯ－３为１８７．６±７３．５μｍｏｌ／Ｌ比１２２．４±４２．６μｍｏｌ／Ｌ），心衰组患者血清ＮＯ和ＴＮＦ水平较无心衰组亦显著增高（ＮＯ－２／ＮＯ－３为１９８．４±６７．４μｍｏｌ／Ｌ与１４２．６±５７．８μｍｏｌ／Ｌ比；血清ＴＮＦ含量与ＮＯ含量呈显著正相关（ｒ＝０．５７，Ｐ＜０．００５），动脉血氧分压与ＴＮＦ水平呈明显负相关（ｒ＝０．６４，Ｐ＜０．００１）。结论：ＮＯ和ＴＮＦ是参与肺心病发病过程的重要因子，监测血清ＮＯ及ＴＮＦ水平可作为反映肺心病病情变化，指导临床治疗的参考指标。     

哮喘患儿白细胞介素4、γ干扰素水平与IgE的相关性研究

目的了解白细胞介素４（ＩＬ－４）、γ干扰素（ＩＦＮγ）对哮喘患儿ＩｇＥ生成的调节作用。方法应用酶联免疫吸附试验，检测了２２例过敏性哮喘患儿外周血单个核细胞（ＰＢＭＣ）培养上清液中ＩＬ－４和ＩＦＮγ的含量以及血清中ＩＬ－４、ＩＦＮγ和ＩｇＥ的水平，并进行了相关性研究。结果哮喘组患儿ＰＢＭＣ培养上清液中ＩＬ－４与ＩＦＮγ含量分别为２６７．０±１８８．７ｎｇ／Ｌ和０．９７±０．５１μｇ／Ｌ，与对照组（９２．７±１４．７ｎｇ／Ｌ和１．７５±０．８８μｇ／Ｌ）比较，差异有非常显著性（Ｐ＜０．００１，Ｐ＜０．０１）。哮喘组血清中ＩＬ－４和ＩｇＥ含量分别为９０．５±５２．６ｎｇ／Ｌ和９１６．０±５２３．２ｋＵ／Ｌ，而对照组分别为３２．２±２３．０ｎｇ／Ｌ和１８６．６±１２７．７ｋＵ／Ｌ（Ｐ＜０．０１，Ｐ＜０．００１）。哮喘患儿血清中ＩＬ－４与ＩｇＥ浓度呈正相关（ｒ＝０．６１），而ＩＦＮγ与ＩｇＥ呈负相关（ｒ＝－０．４９）。结论提示ＩＬ－４在哮喘患儿ＩｇＥ的产生中起促进作用，而ＩＦＮγ则起抑制作用。     

兔烧伤后血液中性粒细胞粘附及磷脂酶A2改变的研究

揭示烧伤早期机体细胞分子水平的病理变化，为严重烧伤患者的早期救治提供参考，采用兔３０％体表面积烧伤，在伤后即刻，２，４，８，１６，２４和４８小时取血分离中性粒细胞（ＰＭＮ），用ＥＬＩＳＡ法测定不同时间内血中ＰＭＮ粘附蛋白与粘附率的变化及其关系；同时用放免法测定血浆中磷脂酶Ａ２（ＰＬＡ２）的改变，寻找其与ＰＭＮ粘附的内在联系。结果：烧伤后ＰＭＮ粘附蛋白及粘附率均明显高于烧伤前，尤以烧伤后８小时为最显著，然后逐渐下降，至４８小时后ＰＭＮ粘附蛋白和粘附率恢复至烧伤前水平，而兔烧伤后ＰＭＮ粘附蛋白与粘附率两者之间呈明显的正相关（ｒ＝０．９０），说明烧伤后ＰＭＮ粘附蛋白水平的增高是ＰＭＮ粘附率增强的主要原因之一；烧伤后血浆中ＰＬＡ２也开始升高，２小时为最明显，推测烧伤后ＰＬＡ２的变化可能也是ＰＭＮ粘附性改变的原因之一。研究结果为临床烧伤患者病程发展的预测和早期治疗提供理论依据     

Quality control in neutrophil granulocyte (PMN) concentrates by flow cytometry.

BACKGROUND: In peripheral blood, chemotaxis, phagocytosis, and oxidative burst of polymorphonuclear cells (PMNs) can be assessed by flow cytometry, whereas function tests, i.e., quality control in PMN concentrates designed for neutropenia therapy, are lacking. METHODS: PMN concentrates (n=6) harvested from healthy donors who had been premedicated with granulocyte colony-stimulating factor (G-CSF) and dexamethasone were stored undiluted (control, C; n=6) and diluted 1:4 (D; n=6) with autologous plasma for 72 h. Commercial flow cytometry function tests were performed to quantify changes in chemotaxis, phagocytosis, and oxidative burst of PMNs over time. RESULTS: Median levels of phagocytosis and oxidative burst levelled at 86% (82-94) and 98% (83-100) in C on the day of apheresis, respectively, but deteriorated to 15% (0-24) and 0% within 72 h; in D these parameters remained close to 90%. Median levels of chemotaxis were comparable in C (69%, 65-74) and D (74%, 70-84) at baseline. No migration was detected in C after 72 h; however, D retained approximately 63% (13-76) migration capacity. CONCLUSION: Quality control in PMN concentrates is practical using flow cytometry and commercial test kits. While phagocytosis and oxidative burst may be maintained for 72 h in vitro, chemotaxis of apheresed PMNs is already reduced on the day of apheresis.     

A regulatory role for Src homology 2 domain-containing inositol 5'-phosphatase (SHIP) in phagocytosis mediated by Fc gamma receptors and complement receptor 3 (alpha(M)beta(2); CD11b/CD18)

The Src homology 2 domain-containing inositol 5'-phosphatase (SHIP) is recruited to immunoreceptor tyrosine-based inhibition motif (ITIM)-containing proteins, thereby suppressing phosphatidylinositol 3-kinase (PI 3-kinase)-dependent pathways. The role of SHIP in phagocytosis, a PI 3-kinase-dependent pathway, is unknown. Overexpression of SHIP in macrophages led to an inhibition of phagocytosis mediated by receptors for the Fc portion of IgG (Fc gamma Rs). In contrast, macrophages expressing catalytically inactive SHIP or lacking SHIP expression demonstrated enhanced phagocytosis. To determine whether SHIP regulates phagocytosis mediated by receptors that are not known to recruit ITIMs, we determined the effect of SHIP expression on complement receptor 3 (CR3; CD11b/CD18; alpha(M)beta(2))-dependent phagocytosis. Macrophages overexpressing SHIP demonstrated impaired CR3-mediated phagocytosis, whereas macrophages expressing catalytically inactive SHIP demonstrated enhanced phagocytosis. CR3-mediated phagocytosis in macrophages derived from SHIP(-/-) mice was up to 2.5 times as efficient as that observed in macrophages derived from littermate controls. SHIP was localized to Fc gamma R- and CR3-containing phagocytic cups and was recruited to the cytoskeleton upon clustering of CR3. In a transfected COS cell model of activation-independent CR3-mediated phagocytosis, catalytically active but not inactive SHIP also inhibited phagocytosis. We conclude that PI 3-kinase(s) and SHIP regulate multiple forms of phagocytosis and that endogenous SHIP plays a role in modulating beta(2) integrin outside-in signaling.     

Flow cytometric and optical microscopic evaluation of poly(D, L-lactide-co-glycolide) microspheres phagocytosis by pig alveolar macrophages.

The phagocytosis of fluorescent poly(D,L-lactide-co-glycolide) microspheres by fresh and frozen pig alveolar macrophages was investigated by optical microscopy on adherent cell culture and by flow cytometry with cell suspension. The kinetic of phagocytosis was studied on a 360 min period as a function of the ratio between microspheres and macrophages (MS:AM ratio from 1:1 to 10:1). No difference of phagocytosis between fresh and frozen macrophages was observed whatever the MS:AM ratio following flow cytometric evaluation while a significant phagocytosis pattern was noticed following optical microscopic evaluation for the highest ratio. The intensity of phagocytosis was dependent on the duration of incubation and dependent, but not proportionally, to the MS:AM ratio showing that the highest efficiency was obtained with the MS:AM ratio of 1:1. Flow cytometry analysis has shown a correlation between cell population and fluorescent events suggesting that phagocytosis of nonfluorescent antigen-loaded particles with different characteristics could be investigated.     

High-affinity, human antibody-like antibody fragment (single-chain variable fragment) neutralizing the lethal factor (LF) of Bacillus anthracis by inhibiting protective antigen-LF complex formation

The anthrax lethal toxin (LT) consists of two subunits, the protective antigen (PA) and the lethal factor (LF), and is essential for anthrax pathogenesis. Several recombinant antibodies directed against PA and intended for medical use have been obtained, but none against LF, despite the recommendations of anthrax experts. Here we describe an anti-LF single-chain variable fragment (scFv) that originated from an immunized macaque (Macaca fascicularis) and was obtained by phage display. Panning of the library of 1.8 x 10(8) clones allowed the isolation of 2LF, a high-affinity (equilibrium dissociation constant, 1.02 nM) scFv, which is highly neutralizing in the standardized in vitro assay (50% inhibitory concentration, 1.20 +/- 0.06 nM) and in an in vivo assay. The scFv neutralizes anthrax LT by inhibiting the formation of the LF-PA complex. The genes encoding 2LF are very similar to those of human immunoglobulin germ line genes, sharing substantial (84.2%) identity with their most similar, germinally encoded counterparts; this feature favors medical applications. These results, and others formerly published, demonstrate that our approach can generate antibody fragments suitable for prophylaxis and therapeutics.     

Empirical mode decomposition analysis of HRV data from patients undergoing local anaesthesia (brachial plexus block)

Spectral analysis of heart rate variability (HRV) is used for the assessment of cardiovascular autonomic control. In this study, a data-driven adaptive technique called empirical mode decomposition (EMD) and the associated Hilbert spectrum has been used to evaluate the effect of local anaesthesia on HRV parameters in a group of 14 patients undergoing axillary brachial plexus block. The normalized amplitude Hilbert spectrum was used to calculate the error index associated with the instantaneous frequency. The amplitude and the frequency values were corrected in the region where the error was higher than twice standard deviation. The intrinsic mode function (IMF) components were assigned to the LF and the HF part of the signal by making use of the centre frequency and the standard deviation spectral extension estimated from the marginal spectrum of the IMF components. The optimal range of the stopping criterion was found to be between 4 and 9 for the HRV data. The statistical analysis showed that the LF/HF ratio decreased within an hour of the application of the brachial plexus block compared to the values at the start of the procedure. These changes were observed in 13 of the 14 patients included in this study.     

Effect of spontaneous saliva swallowing on short‐term heart rate variability (HRV) and reliability of HRV analysis

The effects of effortful swallowing and solid meal ingestions on heart rate variability (HRV) have been examined previously. The effects of spontaneous saliva swallowing on short‐term HRV and reliability of HRV analysis have not been studied before. The effect of saliva swallowing on HRV analyses parameters [meanRRI, SDNN (standard deviation of normal‐to‐normal), LF (low frequency), HF (high frequency) powers, LH/HF] and the reliability of LF and HF powers were investigated by frequency, time–frequency and intraclass correlation coefficient (ICC) analyses. Electrocardiogram and swallowing signal that obtained from an electronic stethoscope placed on the necks of subjects were recorded simultaneously from 30 healthy and young volunteers in sitting position during 15 min. Spontaneous swallowing has been shown to significantly alter some HRV parameters (SDNN, LF power and LF/HF ratio). Time‐frequency analysis results showed that the contribution of saliva swallowing to LF (1–58%) and HF (2–42%) powers could change significantly depending on the number of swallowing. The ICC of the LF and HF powers for the successive 5‐min signal segments were found 0·89, 0·92, respectively. These values decreased to 0·73 and 0·90 in the subjects with more swallowing rate. When the analyses were made for 2‐min signal periods, these values decreased to 0·63 and 0·67. We concluded that spontaneous saliva swallowing can change HRV parameters. We have also seen that changes in swallowing rate and use of short signal segments may reduce the reliability of HRV analyses.     

Leukocytic thermostable alpha-glycoprotein and lactoferrin from granulocyte cytoplasm (immunocytochemical study)

The immunofluorescence techniques with monospecific antibodies to lactoferrin (LF) and leukocytic thermostable alpha-glycoprotein (LT alpha-G) has demonstrated that both the proteins are specific components of the peripheral blood granulocytes of normal subjects. LF and LT alpha-G are detectable in the cytoplasmic granules of polynuclear leukocytes but not on the external surface of these cells' membranes. Differences between the direct and indirect immunofluorescence techniques, used for the detection of the granulocytic LF and LT alpha-G are discussed. Examinations of the blood smears from patients, operated on under artificial circulation, have shown that the emergence of granulocytic protein aggregates between the cells and LF and LT alpha-G sorption on the red cell surface indicate a disease.     

Immunomodulatory effects of Premna tomentosa (L. Verbenaceae) extract in J 779 macrophage cell cultures under chromate (VI)-induced immunosuppression

OBJECTIVE: In the present study, the immunomodulatory effects of Premna tomentosa extract against chromate (VI)-induced toxicity was assessed in J 779 macrophage cell line. DESIGN: The cells were analyzed for cytotoxicity, phagocytosis, oxidant burst, antioxidant status, and cell proliferation. RESULT: Chromate treatment resulted in a significant increase in cytotoxicity and free radical production. Furthermore, there is a significant decrease in reduced glutathione (GSH) levels and glutathione peroxidase activity (GPx). There was an appreciable decrease in cell proliferation and phagocytosis by macrophages in the presence of chromate. However, pretreatment of the cells with P. tomentosa extract (500 microg concentration), 30 minutes prior to chromate (VI) treatment resulted in a significant inhibition of chromate-induced cytotoxicity and reactive oxygen species production. The extract also restored the antioxidant status, cell proliferation, and phagocytosis similar to that of control cells. CONCLUSION: The results confirm the cytoprotective and immunomodulatory effects of the leaves of P. tomentosa and its possible usage in immunosuppressed conditions.     

The effect of an NADH oxidase inhibitor (hydrocortisone) on polymorphonuclear leukocyte bactericidal activity

Polymorphonuclear neutrophils (PMN) from patients with chronic granulomatous disease of childhood have impaired bactericidal activity and are deficient in diphosphopyridine nucleotide, reduced form of, (NADH) oxidase. Since hydrocortisone had been shown to inhibit NADH oxidation, experiments were undertaken to determine the effect of hydrocortisone on several parameters of human PMN function.The phagocytic and bactericidal capacity of PMN with or without hydrocortisone (2.1 mM) was determined by quantitation of cell-free, cell-associated, and total bacteria. Phagocytosis of Staphylococcus aureus and several gram-negative rods was unimpaired by the presence of hydrocortisone in the media. In contrast, killing of bacteria was markedly impaired by hydrocortisone. After 30 min of incubation, there were 20-400 times as many bacteria surviving in hydrocortisone-treated PMN as in simultaneously run controls without hydrocortisone.The defect of intracellular killing noted in the presence of hydrocortisone was not related to impaired degranulation. Quantitative kinetic studies of degranulation revealed no difference in the release of granule associated acid phosphatase in hydrocortisone-treated and control PMN after phagocytosis. Electron microscopy of PMN also indicated that the presence of hydrocortisone had no effect on the extent of degranulation after phagocytosis. These observations were confirmed by studies using histochemical techniques to detect lysosomal enzymes.After phagocytosis, hydrocortisone-treated PMN demonstrated less NADH oxidase activity, oxygen consumption, and hydrogen peroxide production than postphagocytic control PMN. In addition, Nitro blue tetrazolium dye reduction was diminished in hydrocortisone-treated PMN.Thus, impairment of NADH oxidase activity in normal human PMN by hydrocortisone results in reduced intracellular killing of bacteria, diminished postphagocytic oxygen consumption, decreased ability to reduce Nitro blue tetrazolium, and decreased hydrogen peroxide production. These abnormalities are similar to those seen in the PMN of patients with chronic granulomatous disease of childhood.     

房室传导间期(A-V)的自动测量及频谱特征

目的 探讨房室传导间期(A-V)的自动测量,频谱特点及心率对其影响。方法 实验是在去神经的猫上进行,由右颈总动脉插入主动脉根部,测出希氏束(His)电图,通过模板匹配的方法自动检测His电图中心房(A)、希氏(H)波及心室(V)波,并将AA、AV间期数据通过快速傅立叶(FFT)转换,获得其频谱特征。再通过程序控制心房起搏观察了AA间期变化对AV频谱的影响。结果His束中A、H、V波可以自动重复检测,AV频谱与AA频谱均相似,但密度(PSD)较AA小,经标准化后,两者的高频(HF),低频(LF)及高频/低频(HF/LF)相同(P>0.05),通过心房起搏表明AA间期与AV间期呈非线形关系,AV频谱中HF与AA间期的变化率呈负相关(R=-0.97,P<0.01),LF与AA间期的变化率呈正相关关系(R=0.96,P<0.01)。结论心脏His束自动测量揭示了房室传导的频域特征以及心率对AV频谱各成份的相关关系,为研究心脏房室传导障碍性疾病以及心率对心电频谱的影响提供了一个新的方法。