中性粒细胞外陷阱在血栓形成中的作用及研究进展

血栓性疾病严重危害人群健康。中性粒细胞外陷阱作为中性粒细胞的固有免疫机制之一,在帮助机体抵御病原体之外还可通过其结构的支架作用和组分的凝血激活作用促进血栓形成,并在动静脉血栓、抗凝脂综合征以及肿瘤相关血栓的动物疾病模型、人类血栓样本和体外实验中得到了证实。主要介绍中性粒细胞外陷阱的形成机制,重点介绍中性粒细胞外陷阱与血栓形成的关联,以及在血栓性疾病和新型冠状病毒肺炎中作用,旨在为血栓防治相关研究提供参考。     

Neutrophil to lymphocyte and platelet to lymphocyte ratios in systemic lupus erythematosus: Relation with disease activity and lupus nephritis

ObjectivesTo investigate the role of neutrophil to lymphocyte ratio (NLR) and platelet to lymphocyte ratio (PLR) as activity markers in systemic lupus erythematosus (SLE) without nephritis and lupus nephritis (LN) patients.Patients and methodsThis study included 60 SLE patients with LN, 60 SLE patients without renal involvement and 30 healthy controls. We analyzed correlations between NLR and PLR and both disease activity and renal affection.ResultsThe NLR of SLE patients was much higher than those of the controls. Both ratios showed significantly increased values in SLE patients with active disease. NLR and PLR were positively correlated with SLEDAI, ESR, and CRP and negatively correlated with C4. SLE patients with LN had higher levels of NLR than those without nephritis. NLR showed positive correlations with BUN, serum urea, serum creatinine and 24 h urinary protein. We found NLR to be related to anti-ds-DNA level and renal biopsy classes. While PLR was related only to anti ds-DNA. The best NLR to predict SLE active disease was 2.2 and the best PLR cut-off value was 132.9.ConclusionNLR and PLR are useful inflammatory markers to evaluate disease activity in SLE patients. Also, NLR could reflect renal involvement in SLE patients and is associated with the different classes of its histological staging.     

Involvement of neutrophil gelatinase-associated lipocalin and osteopontin in renal tubular regeneration and interstitial fibrosis after cisplatin-induced renal failure

The kidney has a capacity to recover from ischemic or toxic insults that result in cell death, and timely tissue repair of affected renal tubules may arrest progression of injury, leading to regression of injury and paving the way for recovery. To investigate the roles of neutrophil gelatinase-associated lipocalin (NGAL/lcn2) and osteopontin (OPN/spp1) during renal regeneration, the expression patterns of NGAL and OPN in the cisplatin-induced rat renal failure model were examined. NGAL expression was increased from day 1 after injection; it was seen mainly in the completely regenerating proximal tubules of the cortico-medullary junction on days 3–35; however, the expression was not seen in abnormally dilated or atrophied renal tubules surrounded by fibrotic lesions. On the other hand, OPN expression was increased from day 5 and the increased expression developed exclusively in the abnormal renal tubules. NGAL expression level well correlated with the proliferating activity in the regenerating renal epithelial cells, whereas OPN significantly correlated with the α-smooth muscle actin-positive myofibroblast appearance, expression of transforming growth factor (TGF)-β1, and the number of CD68-positive macrophages. Interestingly, rat renal epithelial cell line (NRK-52E) treated with TGF-β1 decreased NGAL expression, but increased OPN expression in a dose-dependent manner. Because increases of TGF-β1, myofibroblasts and macrophages contribute to progressive interstitial renal fibrosis, OPN may be involved in the pathogenesis of fibrosis; on the contrary, NGAL may play a role in tubular regeneration after injury. Expression analysis of NGAL and OPN would be useful to investigate the tubule damage in renal-toxicity.     

不同剂量阿托伐他汀对经皮冠状动脉介入术后对比剂急性肾损伤的影响

目的 观察经皮冠状动脉介入术（PCI）前使用不同剂量阿托伐他汀对对比剂诱导的急性肾损伤的影响。方法 入选拟行冠状动脉造影检查及拟行PCI术的患者106例,随机分成2组：20 mg阿托伐他汀组及40 mg阿托伐他汀组;入院24 h内完成常规化验检查、心脏彩色超声检查及肾脏血管超声检查,术后48 h复查肾功能。所有患者手术前当日清晨、术后2 h及术后48 h均留取约5 mL中段尿,用胶乳增强免疫透射比浊法统一测定中性粒细胞明胶酶相关脂质运载蛋白（NGAL）。结果 PCI术后,40 mg阿托伐他汀组与20 mg阿托伐他汀组比较,血清肌酐（77.44±23.14 mmol/L比94.24±36.14 mmol/L,P＝0.014）降低,尿酸（313.05±110.84 μmol/L比354.00±100.66 μmol/L,P＝0.060）降低,肾小球滤过率估计值（92.24±24.74比75.31±31.34,P＝0.009）增高; PCI术后,40 mg阿托伐他汀组与20 mg阿托伐他汀组比较,2 h NGAL（33.13±20.44 μg/L比50.67±46.95 μg/L,P＝0.013）、48 h NGAL（27.56±18.64 μg/L比58.38±56.81 μg/L,P＝0.001）减低;应用对比剂后,20 mg阿托伐他汀组发生对比剂急性肾损伤11例,发生率为20.75%,而40 mg阿托伐他汀组发生对比剂急性肾损伤5例,发生率为9.43%,两组相比差别有统计学意义（P<0.05）。结论 应用对比剂前3天,每天服用40 mg阿托伐他汀较每天服用20 mg阿托伐他汀更能减少对比剂诱导的急性肾损伤的发生。     

银黄口服液联合头孢唑林钠治疗老年急性上呼吸道感染的临床研究

目的评价银黄口服液联合头孢唑林钠治疗老年急性上呼吸道感染的临床疗效。方法选取2016年9月—2017年9月四川大学华西医院收治的161例老年急性上呼吸道感染患者,根据用药差别分成对照组(80例)和治疗组(81例)。对照组患者静脉滴注注射用头孢唑林钠,1.0 g注射用头孢唑林钠溶于250 mL生理盐水,2次/d。治疗组患者在对照组治疗基础上口服银黄口服液,10 mL/次,3次/d。两组患者均连续治疗7 d。观察两组的临床疗效,比较两组患者临床症状消失时间、症状评分、血清鳞状细胞癌抗原(SCC)、超敏C反应蛋白(hs-CRP)及白细胞百分比的情况。结果治疗后,对照组和治疗组的总有效率分别是86.25%、96.30%,两组比较差异具有统计学意义(P0.05)。治疗后,两组患者血清SCC、hs-CRP及淋巴细胞百分比水平均较治疗前明显降低,同组治疗前后比较差异有统计学意义(P0.05);治疗后治疗组SCC、hs-CRP及淋巴细胞百分比显著低于对照组,两组比较差异具有统计学意义(P0.05)。治疗后,治疗组患者退热时间、咽喉肿胀消退时间均明显短于对照组,两组比较差异具有统计学意义(P0.05)。治疗后,两组患者症状积分均较治疗前显著降低,同组治疗前后比较差异有统计学意义(P0.05);治疗后治疗组症状积分显著低于对照组,两组比较差异具有统计学意义(P0.05)。结论银黄口服液联合头孢唑林钠治疗老年急性上呼吸道感染具有较好的临床疗效,能显著改善患者临床症状,减轻患者炎症程度,具有一定的临床推广应用价值。     

Target-mediated clearance and bio-distribution of a monoclonal antibody against the Kunitz–type protease inhibitor 2 domain of Tissue Factor Pathway Inhibitor

Introduction

A humanised monoclonal antibody, concizumab, that binds with high affinity to the Kunitz-type protease inhibitor (KPI) 2 domain of human tissue factor pathway inhibitor (TFPI) is in clinical development. It promotes coagulation by neutralising the inhibitory function of TFPI and may provide a subcutaneous prophylaxis option for patients with haemophilia. We aimed to study biodistribution and pharmacokinetics (PK) of concizumab.

Materials and Methods

Blockage of cellular TFPI by concizumab was measured by tissue factor/Factor VIIa-mediated Factor X activation on human EA.hy926 cells. Biodistribution of concizumab was analysed in rabbits by immunohistology, and the PK was measured in rabbits and rats.

Results and Conclusions

Concizumab bound to cell surface TFPI on EA.hy926 cells and neutralised TFPI inhibition of Factor X activation. The antibody cross-reacted with rabbit TFPI, but not with rat TFPI, allowing for comparative PK studies. PK data in rats described a log-linear profile typical for a non-binding antibody, whereas PK data in rabbits revealed a non-linear, dose-dependent profile, consistent with a target-mediated clearance mechanism. Immunohistology in rabbits during target-saturation showed localisation of the antibody on the endothelium of the microvasculature in several organs. We observed a marked co-localisation with endogenous rabbit TFPI, but a negligible sub-endothelial build-up. Concizumab binds and neutralises the inhibitory effect of cell surface-bound TFPI. The PK profile observed in rabbits is consistent with a TFPI-mediated drug disposition. Double immunofluorescence shows co-localisation of the antibody with TFPI on the endothelium of the microvasculature and points to this TFPI as a putative target involved in the clearance mechanism.     

The lowest VE/VCO2 ratio best identifies chronic thromboembolic pulmonary hypertension

Introduction

The natural history of acute pulmonary embolism (PE) under treatment is about a gradual resolution of the thrombi, and uncommonly, the development of chronic thromboembolic pulmonary hypertension (CTEPH). We hypothesized that ventilatory efficiency parameters during cardiopulmonary exercise testing (CPET) may be able to monitor the process and predict CTEPH.

Methods

15 patients rehabilitated from acute PE (total resolution of thrombi), 44 patients with chronic PE (with residual thrombi), 66 patients with CTEPH, and 36 sedentary healthy controls performed incremental CPET.

Results

The lowest VE/VCO₂ was higher in CTEPH patients than that in chronic PE and rehabilitated patients (43.4 L/min vs 29.9 L/min vs 27.1 L/min, p < 0.005). The VE/VCO₂ slope (48.4 L/min/L/min vs 29.9 L/min/L/min vs 28.0 L/min/L/min, p < 0.005) and oxygen uptake efficiency plateau (OUEP) (37.1 L/min vs 27.0 L/min vs 25.2 L/min, p < 0.005) had the similar changes. In logistic regression analysis, the lowest VE/VCO₂ ≥ 34.35 L/min was the best predictor of CTEPH (OR 159.0, 95% CI 36.0-702.3, p < 0.001). The lowest VE/VCO₂ was higher in chronic PE patients compared with the controls (29.9 L/min vs 26.5 L/min, p < 0.05), but there was no difference between the rehabilitated patients and the controls. In multiple linear regression analysis, the percentage of vascular obstruction by ventilation-perfusion lung scanning (PVO) was the most significant independent predictor for indices of ventilatory efficiency in chronic PE and rehabilitated patients.

Conclusions

CTEPH is associated with weakened ventilatory efficiency. The lowest VE/VCO2 ratio has the best capability to predict CTEPH. Ventilatory inefficiency improves along with recovery of acute PE.     

Rac1在小鼠中性粒细胞抗白假丝酵母菌感染中的作用

目的评价Rac1基因在小鼠抗白假丝酵母菌感染中的作用。方法 (1)β-actin作为内参,Western blot半定量法检测两种小鼠中性粒细胞Rac1蛋白表达量;(2)趋化实验用Zigmond法经甲酰甲硫氨酰-亮氨酰-苯丙氨酸(fMLP)诱导中性粒细胞移动,延时显微镜观察中性粒细胞移动并拍照,细胞追踪软件分析小鼠中性粒细胞趋化功能;(3)fMLP诱导小鼠中性粒细胞过氧化物的产生应用异氨基苯二酰肼化学发光分析法通过微孔板发光检测仪检测过氧化物酶含量,采用细胞色素c还原法分析豆蔻酸-佛波醇-乙酸酯(PMA)引发中性粒细胞产生的过氧化物。结果假丝酵母菌耐受BALB/c小鼠中性粒细胞Rac1蛋白表达量高于假丝酵母菌易感CBA/CaH小鼠;BALB/c小鼠中性粒细胞趋化运动功能较CBA/CaH小鼠强(P<0.05);经fMLP和PMA诱导后,BALB/c小鼠中性粒细胞过氧化物的产生量高于CBA/CaH小鼠(P<0.05)。结论在小鼠中性粒细胞抗白假丝酵母菌感染过程中,Rac1能增强小鼠中性粒细胞的杀伤能力。     

牙周炎与慢性阻塞性肺疾病关系的流行病学研究

目的通过流行病学调查及临床检查,探讨牙周炎与慢性阻塞性肺疾病（COPD）之间的相关性。方法收集中国医科大学附属第一医院、盛京医院、第四医院和沈阳市第八人民医院2008年10月至2009年4月确诊为COPD的患者266例,其中稳定期COPD患者160例,急性加重期COPD（AE—COPD）患者106例,检查并记录所有患者6颗指数牙（61 6、6 16）的简化1：7腔卫生指数（OHI—S）、龈沟出血指数（SBI）、牙周探诊深度（PD）、临床附着丧失（CAL）,同时对所有患者进行肺功能指标的检查,并进行口腔问卷调查。结果（1）AE—COPD组的吸烟率为47．2％,明显高于稳定期COPD组（P〈0．01）;（2）AE—COPD组的SBI、PD及CAL明显高于稳定期COPD组（P〈0．01）;（3）AE—COPD组的OHI—S、SBI、PD及CAL均与FEVl％呈负相关（r分别为-0．309、-0．333、-0．395、-0．702,均P〈0．01）,其中CAL与FEVI％的相关性最强;稳定期COPD组CAL与FEVI％呈负相关（r=-0．657,P〈0．01）。结论（1）吸烟作为牙周炎和COPD的共同危险因素,在牙周炎和COPD的发生和发展中发挥重要作用;（2）AE—COPD患者的牙周炎症程度均随着肺功能的减弱而加重,提示牙周炎与COPD之间存在一定相关性。