雷公藤红素对人中性粒细胞吞噬功能的实验研究

目的 研究雷公藤红素对人中性粒细胞mTOR mRNA表达、吞噬功能的影响,探索其对中性粒细胞相关免疫功能可能产生的影响.方法 分离健康人外周血中性粒细胞,不同浓度雷公藤红素孵育不同时间,RT_PCR检测mTORmRNA表达量,同时采用预染色金黄色葡萄球菌法评价中性粒细胞吞噬功能.结果 雷公藤红素能够下调mTOR表达,抑制粒细胞吞噬功能,与浓度呈负相关性.结论 雷公藤红素可能通过下调mTOR mRNA表达,影响中性粒细胞吞噬功能.     

瘢痕子宫产妇中性粒细胞功能变化研究

目的：探讨瘢痕子宫产妇中性粒细胞功能变化。方法选取2011年5月—2012年9月开滦总医院产科收治的瘢痕子宫产妇108例作为瘢痕子宫组,另选取同期开滦总医院产科收治的非瘢痕子宫产妇126例作为对照组。观察两组中性粒细胞趋化活性（中性粒细胞趋化距离、中性粒细胞趋化指数）及中性粒细胞吞噬功能（中性粒细胞吞噬百分率、中性粒细胞吞噬指数）。结果瘢痕子宫组中性粒细胞趋化距离长于对照组,中性粒细胞趋化指数高于对照组,差异有统计学意义（P﹤0.05）;两组中性粒细胞吞噬百分率比较,差异无统计学意义（P﹥0.05）,瘢痕子宫组中性粒细胞吞噬指数低于对照组,差异有统计学意义（ P﹤0.05）。结论瘢痕子宫的形成可能与中性粒细胞趋化活性升高、吞噬功能降低有关。     

2型糖尿病患者外周血白细胞计数与尿白蛋白相关性的分析

目的探讨2型糖尿病患者外周血白细胞计数与尿白蛋白相关性情况。方法选取笔者所在医院2型糖尿病患者90例作为观察组,同时选取同期健康体检者100例作为对照组。结果两组肝功、肾功能差异无统计学意义(P>0.05)。观察组血糖明显高于对照组,观察组白细胞总数、中性粒细胞、单核细胞、嗜酸性粒细胞、嗜碱性粒细胞和尿白蛋白均明显高于对照组,而淋巴细胞明显低于健康对照组,2型糖尿病患者尿白蛋白和白细胞总数、中性粒细胞、单核细胞、嗜酸性粒细胞、嗜碱性粒细胞呈明显正相关,而与淋巴细胞呈负相关,差异有统计学意义(P<0.05)。结论 2型糖尿病患者外周血白细胞计数和尿白蛋白具有明显的相关性,其可能和糖尿病的肾脏损伤密切相关。     

对冠心病患者外周血T淋巴细胞亚群及中性粒细胞功能的观察

免疫机理异常在冠心病发生发展中的作用已引起学者们的关注,但迄今对T淋巴细胞亚群在冠心病,尤其在陈旧性心梗中的改变,报道甚少,国内尚未见关于本病中性粒细胞功能改变的临床观察。我们用单克隆抗体免疫花环法(直接法)测定了本病患者外周血T淋巴细胞亚群,并同时测定了中性粒细胞的吞噬和杀伤功能,结果见表1及表2。     

青霉素G对人类单核细胞吞噬的金黄色葡萄球菌的作用

目前认为治疗细菌性感染,抗生素对吞噬细菌的作用非常重要。抗生素能否渗入细胞内杀伤细菌,不少学者已经用粒细胞作过研究,并证明了青霉素 G 在粒细胞内没有活性。但至今未见青霉素 G 对单核细胞吞噬的金黄色葡萄球菌的作用的报道。为此,作者进行了如下研究。作者从健康自愿者血液中分离单核细胞。用每毫升含0.5单位的肝素洗四次,再用HBSS 把单核细胞稀释成10~7浓度的悬液。金黄色葡萄球菌(42D 型)用含10%的AB 型血清悬液,在37℃孵育30分钟,而后将细菌在1500g 离心10分钟,用冰冷的 HBSS明胶洗两次后,用 HBSS 明胶把金黄色葡萄球菌稀释为每毫升含10~7的菌液。检测青霉素 G 对吞噬细胞吞噬金黄色葡萄球菌的作用。将每毫升含10~7预先处理的金黄色葡萄球菌和每毫升含10~7的单核细胞一起,37℃孵育3分钟后,在冰浴中冷却迅     

欧瑞血活素注射液对小鼠免疫功能的影响

目的:观察欧瑞血活素注射液对小鼠免疫功能的影响。方法:以腹腔注射方式给予昆明小鼠不同剂量的欧瑞血活素注射液30d ,并以生理盐水腹腔注射为阴性对照,干扰素腹腔注射为阳性对照。结果:欧瑞血活素注射液小、中剂量组均具有提高中性粒细胞的杀伤功能(P<0.05,P<0.01) ;同等剂量组还可以显著地提高小鼠溶血素水平(P<0.05) ;小、中、大剂量组均具有提高碳粒廓清指数和吞噬指数的作用、显著地增加脾脏及胸腺的重量,其中以中剂量组增强单核巨噬细胞的吞噬功能、大剂量组增加脾脏重量、中剂量组增加胸腺重量的作用最为显著。结论:小、中剂量欧瑞血活素注射液可以提高昆明小鼠的特异性免疫、非特异性免疫以及中性粒细胞的吞噬功能。     

白细胞介素-15对白色念珠菌播散的抑制作用

目的探讨白细胞介素 15 (IL 15 )对白色念珠菌播散的作用。方法以小鼠为动物模型 ,检测IL 15对中性粒细胞功能、白色念珠菌黏附组织细胞作用以及血管内白色念珠菌播散的影响。结果IL 15在体外可明显增强中性粒细胞的吞噬和杀菌活性 (P <0 .0 5 ) ,且呈浓度依赖性 ;在体内可明显降低白色念珠菌对细胞的黏附作用 ,从而阻止其散播。结论IL 15对白色念珠菌播散的抑制作用 ,可用于全身性白色念珠菌感染的治疗     

硝苯吡啶对冠心病患者中性粒细胞氧自由基的影响

以鲁米诺(Luminol)依赖的化学发光法(Chemilumineseence,CL)观察硝苯吡啶对冠心病患者中性粒细胞(Polymorphonuclear Leukocyte,PMN)氧化代谢功能的影响.研究对象包括冠心病未服用硝苯吡啶者24例;服用硝苯吡啶者26例,正常对照者30例。结果表明:冠心病服药组中性粒细胞化学发光强度明显受抑,与冠心病未服药组相比,PMN-CL的本底、峰值及吞噬指数明显减低,与正常对照组相比,PMN-CL的峰值及吞噬指数无差异;而未服药组PMN-CL的本底、峰值及吞噬指数明显高于正常对照组。     

脑卒中患者早期血白细胞检测的临床意义

为探讨脑卒中患者早期外周血白细胞(WBC)计数的变化及临床意义,我们对177例脑出血、260例脑梗死患者发病早期外周血WBC计数及中性粒细胞(PMN)结果与431例健康对照者进行回顾性分析,报告如下.     

首次诊断Graves病患者白细胞或中性粒细胞计数减少的相关因素分析

目的 分析首次诊断Graves病患者外周血液中白细胞或中性粒细胞计数减少的相关因素。方法 收集2020年3月～2022年11月在某院就诊的113例Graves病患者一般资料以及实验室检查,根据外周血液中白细胞或中性粒细胞计数是否减少,分为白细胞和中性粒细胞计数正常组（正常组）和白细胞或中性粒细胞计数减少组（减少组）。对首次诊断Graves病患者白细胞或中性粒细胞计数减少的相关因素进行分析。结果 统计分析结果显示,两组患者年龄、性别、病程差异无统计学意义（P> 0.05）,减少组患者游离三碘甲腺原氨酸（FT3）、游离甲状腺素（FT4）、促甲状腺激素受体抗体（TRAb）水平均高于正常组,差异有统计学意义（P <0.05）,两组患者促甲状腺激素（TSH）、甲状腺球蛋白抗体（TGAb）、甲状腺过氧化物酶抗体（TPOAb）水平差异无统计学意义（P> 0.05）。相关分析显示,FT3、FT4、TRAb水平与白细胞或中性粒细胞计数减少呈正相关（P <0.05）。Logistic回归分析显示,高水平TRAb(OR=1.071,95%CI:1.00～1.15)与首次诊断Grave...     

Anidulafungin: an echinocandin antifungal

Anidulafungin (LY-303366, V-echinocandin?, Vicuron Pharmaceuticals, Inc.) is a new echinocandin antifungal agent with broad spectrum activity against Candida and Aspergillus spp. Anidulafungin exhibits low toxicity, concentration-dependent fungicidal activity for Candida, and a prolonged post antifungal effect (> 12h). In vitro activity demonstrates excellent potency and spectrum versus azole-susceptible and -resistant Candida spp. and a low minimum effective concentration for Aspergillus spp. In vivo anidulafungin is fungicidal against Candida in neutropenic animal models of disseminated candidiasis. Against Candida anidulafungin exhibits concentration-dependent killing and clearance of residual fungal burden in target organs (liver, lung, spleen, kidney) and plasma/tissue concentrations exceed the minimum inhibitory and minimum fungicidal concentrations of the infecting organism throughout the dosing interval. Although the activity of anidulafungin in animal models of pulmonary or disseminated aspergillosis shows increased survival and improvement in the pulmonary infarct score, the effect on residual fungal burden and Aspergillus antigenemia determination does not indicate in vivo fungicidal activity. It seems that the major effect of anidulafungin and other echinocandins in vivo against Aspergillus spp. is the decrease in the angioinvasive potential of the organisms. Clinically, anidulafungin has been shown to be safe and effective in the treatment of oesophageal candidiasis and candidemia. Further clinical application of this new antifungal agent is warranted.     

Anidulafungin: an echinocandin antifungal

Anidulafungin (LY-303366, V-echinocandin trade mark, Vicuron Pharmaceuticals, Inc.) is a new echinocandin antifungal agent with broad spectrum activity against Candida and Aspergillus spp. Anidulafungin exhibits low toxicity, concentration-dependent fungicidal activity for Candida, and a prolonged post antifungal effect (> 12h). In vitro activity demonstrates excellent potency and spectrum versus azole-susceptible and -resistant Candida spp. and a low minimum effective concentration for Aspergillus spp. In vivo anidulafungin is fungicidal against Candida in neutropenic animal models of disseminated candidiasis. Against Candida anidulafungin exhibits concentration-dependent killing and clearance of residual fungal burden in target organs (liver, lung, spleen, kidney) and plasma/tissue concentrations exceed the minimum inhibitory and minimum fungicidal concentrations of the infecting organism throughout the dosing interval. Although the activity of anidulafungin in animal models of pulmonary or disseminated aspergillosis shows increased survival and improvement in the pulmonary infarct score, the effect on residual fungal burden and Aspergillus antigenemia determination does not indicate in vivo fungicidal activity. It seems that the major effect of anidulafungin and other echinocandins in vivo against Aspergillus spp. is the decrease in the angioinvasive potential of the organisms. Clinically, anidulafungin has been shown to be safe and effective in the treatment of oesophageal candidiasis and candidemia. Further clinical application of this new antifungal agent is warranted.     

Hydrolysis of kininogens by degranulated human neutrophils and analysis of bradykinin as chemotactic factor for cells isolated from peripheral blood

Human neutrophils play a pivotal role in acute inflammation including the regulation of vascular permeability. We have examined the capacity of neutrophil enzymes to hydrolyse human kininogens in vitro and have also explored the potentiality of bradykinin to induce chemotactic migration on neutrophils isolated from peripheral blood. Isolated neutrophils were stimulated with either f-Met-Leu-Phe, thrombin or silica particles coated with human IgG. Neutrophil enzymes obtained by degranulation produced, after 45 min of incubation with high and low molecular weight kininogens, the complete transformation of both proteins in polypeptides ranging from 20 to less than 10 kDa in molecular mass. Supernatants obtained from nonstimulated neutrophils did not modify the molecular size of kininogens. The assay used to test the chemoattractant capacity of synthetic bradykinin on human neutrophils showed that this peptide has no chemotactic activity on cells isolated from healthy subjects. Our results show that stimulation of human neutrophils with opsonized silica, thrombin and the chemotactic peptide f-Met-Leu-Phe induces release of kininogen-hydrolyzing enzymes from these cells.     

Effects of KW-2228 on the function of polymorphonuclear neutrophils in rats for the phagocytosis and killing activity and the production of active oxygen]

KW-2228 is a novel derivative of a recombinant human granulocyte colony-stimulating factor (rhG-CSF) with a modification. KW-2228 has some excellent properties such as high specific activity in stimulating granulocyte colony-formation in vitro, great biological stability in plasma, good pharmacokinetic profile and high potency in granulopoiesis in normal mice in vivo. In the present study, we investigated the effect of KW-2228 on phagocytic capacity and killing action of polymorphonuclear neutrophils (PMNs) obtained from normal rats which had been treated with KW-2228 using various microorganisms, such as Candida albicans, Escherichia coli and Pseudomonas aeruginosa. We also investigated the effect of KW-2228 on the production of superoxide anion using a luminol-chemiluminescence method. KW-2228 enhanced the phagocytosis and killing activity of PMNs of rats against C. albicans and E. coli. PMNs treated with KW-2228 showed some bactericidal activity against P. aeruginosa. These data obtained with PMNs treated with KW-2228 show a correlation between the bacterial susceptibility to phagocytosis and killing action and the lunimol-dependent chemiluminescence response. These results suggest the significance and the efficacy of KW-2228 used in an additional therapy to that of antibiotics in the treatment of infections diseases. KW-2228 is currently in Phase III clinical trials in Japan.     

Phagocytosis and killing of bacteria and yeast by human milk cells after opsonisation in aqueous phase of milk.

Macrophages and neutrophils from human milk phagocytose and kill Staphylococcus aureus and Escherichia coli in vitro after opsonisation by the aqueous phase of milk as effectively as blood leucocytes in serum. They also phagocytose Candida albicans. The overgrowth of E coli resulting from the addition of iron to cultures of the organism in the aqueous phase of milk is not influenced by the presence of cells. We conclude that the phagocytosis and killing of bacteria by milk cells may contribute to the lower incidence of infection among breast-fed than artificially fed babies.     

骨髓增生异常综合征患者红细胞的天然免疫黏附功能及对自然杀伤细胞杀伤活性的影响

目的 探讨骨髓增生异常综合征患者红细胞天然免疫黏附功能及对自然杀伤细胞（NK细胞）杀伤活性的影响.方法 选择我院骨髓增生异常综合征患者14例为试验组,20例健康人作为对照组,将2组外周血红细胞用柠檬酸钠抗凝,检测红细胞在自身血浆条件下对K562细胞的免疫黏附并计算黏附率;并用四甲基偶氮唑盐微量酶反应比色（MTT）法测定红细胞对正常NK细胞杀伤K562细胞活性的影响,并与未加红细胞时进行比较.结果 试验组红细胞使K562细胞形成了＂玫瑰花＂样结合.对照组红细胞对K562细胞的免疫黏附结合率为（15.6±6.7）%,试验组红细胞对K562细胞的免疫黏附结合率为（8.5±7.0）%,2组比较差异有统计学意义（t=3.66,P＜0.01）.不加红细胞条件下,对照组外周静脉血NK细胞杀伤K562细胞活性杀伤率为64%～75%,加入红细胞后为79%～88%;试验组不加入红细胞为45%～56%,加入红细胞后为51%～58%.加入红细胞后,2组NK红细胞对K562细胞的杀伤率均增加（P＜0.01）,且对照组高于试验组（P＜0.01）.结论 红细胞可增加NK细胞对K562细胞的杀伤率;骨髓增生异常综合征患者的红细胞对K562细胞的免疫黏附能力下降,并可减低NK细胞对K562细胞的杀伤率.     

Neutrophil priming by granulocyte colony stimulating factor and its modulation by protein kinase inhibitors.

Upon stimulation by various ligands, freshly isolated human peripheral neutrophils (PMN) respond in a variety of ways, such as superoxide (O2-.) generation, phagocytosis enzyme release, migration etc. Chemotactic peptide formylmethionyl-leucyl-phenylalanine (FMLP) and opsonized zymosan activate neutrophils by a receptor-mediated mechanism, while phorbol myristate acetate and dioctanoylglycerol activate the cells by a mechanism involving Ca(2+)-and phospholipid-dependent protein kinase (PKC). Receptor-mediated but not PKC-mediated O2-. generation in PMN was enhanced by the priming of recombinant human granulocyte colony stimulating factor (G-CSF). FMLP-dependent luminol chemiluminescence was also enhanced by G-CSF. However, no appreciable enhancement was observed in FMLP-induced intracellular calcium ion concentration ([Ca2+]i). Enhancement of FMLP-induced generation of O2-. by G-CSF was inhibited by genistein or alpha-cyano-3-ethoxy-4-hydroxy-5-phenylthiomethylcinnamamide (ST 638), inhibitors of tyrosine kinase (TK), and was stimulated by staurosporine and 1-(5-isoquinolinesulfonyl)-3-methyl-piperazine (H-7), inhibitors of PKC. The ED50 values of genistein and ST 638 for the inhibition of the FMLP-induced O2-. generation from G-CSF were 0.5 and 5 microM, respectively. In contrast, O2-. generation by PKC activation without G-CSF priming was inhibited by stauroporine and H-7, but was stimulated by genistein and ST 638. These results suggested that the enhancing effect of G-CSF on receptor-mediated generation of the O2-. might be regulated by protein kinases, such as TK and PKC, and that the TK inhibitor selectively inhibited the G-CSF-primed receptor-mediated O2-. generation of neutrophils.     

Superoxide scavenging activity of leukocytes in rheumatoid arthritis and Beh?et's diseases

Superoxide radical anion (O2-) has been suggested to mediate tissue damage associated with chronic inflammatory disorders such as rheumatoid arthritis (RA) and Beh?et's disease (BD). Despite that protection from O2- is provided by superoxide dismutase (SOD), there are controversial results about SOD-like activity of polymorhonuclear cells (PMN) in RA and there are no reports about it in BD. In addition, colchicine, a potent inhibitor of phagocytosis, has been used for the treatment of BD, no report about its effect on SOD-like activity of PMN is available. The present study investigates the superoxide scavenging activity (SSA) of PMN and mononuclear cells (MNC) in both RA and BD, and it examines the effects of colchicine on SSA of PMN in BD using the electron paramagnetic resonance/spin trapping method. The SSA of PMN, but not MNC, was significantly lower in patients with either RA or BD, as compared with that in healthy controls. The SSA of PMN in both RA and BD showed a strong negative correlation with that in healthy controls. The SSA of PMN in both RA and BD showed a strong negative correlation with erythrocyte sedimentation rates and C-reactive protein levels. Colchicine treatment increased the SSA of PMN toward normal level in BD patients, and prevented the decrease of SSA in PMN obtained from healthy adults after the stimulation with opsonized zymosan in vitro. Our results suggest that the decrease of SSA in PMN is not disease specific and it is probably the results of the increased amount of O2- generated by these cells.(ABSTRACT TRUNCATED AT 250 WORDS)     

Fungicidal activity of human lactoferrin-derived peptides based on the antimicrobial αβ region

Owing to the increasing number of infections in hospitalised patients caused by resistant strains of fungi, there is a need to develop new therapeutic agents for these infections. Naturally occurring antimicrobial peptides may constitute models for developing such agents. A modified peptide sequence (CFQWKRAMRKVR; HLopt2) based on amino acid residues 20-31 of the N-terminal end of human lactoferrin (hLF) as well as a double-sized human lactoferricin-like peptide (amino acid residues 16-40; HLBD1) were investigated for their antifungal activities in vitro and in vivo. By in vitro assay, HLopt2 was fungicidal at concentrations of 12.5-25 μg/mL against Cryptococcus neoformans, Candida albicans, Candida krusei, Candida kefyr and Candida parapsilosis, but not against Candida glabrata. HLopt2 was demonstrated to have ≥ 16-fold greater killing activity than HLBD1. By inducing some helical formation caused by lactam bridges or by extending the assay time (from 2h to 20 h), HLBD1 became almost comparable with HLopt2 in its fungicidal activity. Killing of C. albicans yeast cells by HLopt2 was rapid and was accompanied by cytoplasmic and mitochondrial membrane permeabilisation as well as formation of deep pits on the yeast cell surface. In a murine C. albicans skin infection model, atopic treatment with the peptides resulted in significantly reduced yields of Candida from the infected skin areas. The antifungal activities of HLopt2 in vitro and in vivo suggest possible potential as a therapeutic agent against most Candida spp. and C. neoformans. The greatly improved antifungal effect of the lactam-modified HLBD1 indicates the importance of amphipathic helix formation for lethal activity.     

Effect of butylated hydroxyanisole and some of its derivatives on human neutrophil oxidative burst: chemiluminescence evaluation

An acute inflammatory response begins during the reperfusion phase following an ischemic insult in which polymorphonuclear neutrophils (PMNs) play an important role and the release of reactive oxygen species (ROS) causes further damage and a reduction in endogenous antioxidant storage. The ability of butylated hydroxyanisole (BHA) and some phenolic, aliphatic and aromatic BHA derivatives to reduce the human PMN oxidative burst evoked by particulate (Candida albicans and zymosan) or soluble stimulants [N-formyl-methionyl-leucyl-phenylalanine (fMLP) and phorbol myristate acetate (PMA)] was investigated using luminol-amplified chemiluminescence. BHA and the derivative dt-BHA [3,5-di-t-butyl-4-hydroxyanisole] significantly reduced the PMN oxidative burst at concentrations ranging from 5 x 10(-6) to 5 x 10(-5) mol/l for C. albicans stimulation, while for zymosan stimulation, reduction was seen at concentrations ranging from 5 x 10(-6) to 5 x 10(-5) mol/l for BHA, and at concentrations ranging from 5 x 10(-7) to 5 x 10(-5) mol/l for dt-BHA, with dt-BHA being the most active. Another BHA derivative, Bu GAM 1, was active at 5 x 10(-5) mol/l for C. albicans and at 5 x 10(-6) to 5 x 10(-5) mol/l for zymosan. The findings obtained with fMLP and PMA were very similar to those previously reported. ROS release is related to PMN killing activity, but the inhibition of the PMN oxidative burst induced by BHA and BHA derivatives did not significantly modify PMN phagocytosis or killing. It has recently been observed that dt-BHA has a spasmolytic action by inhibiting the influx of Ca(2+) into cells through L-type Ca(2+) channels, which means that a single molecule is capable of counteracting two major steps in the sequence of events triggered by ischemia-reperfusion injury, i.e. free radical release and Ca(2+) overload.