膜结合型肿瘤坏死因子相关凋亡诱导配体修饰的人骨髓基质干细胞对脑胶质瘤细胞的体外靶向抗肿瘤作用

目的：探索经膜结合型肿瘤坏死因子相关凋亡诱导配体（TRAIL）修饰的人骨髓基质干细胞（hMSCs）对脑胶质瘤的靶向抗肿瘤作用。方法：通过体外Transwell系统研究hMSCs向胶质瘤的迁徙能力。采用经全长人TRAIL（hTRAIL）重组的腺相关病毒（rAAV）载体（rAAV hTRAIL）对骨髓基质干细胞（MSCs）进行修饰。将hTRAIL修饰的MSCs与人胶质瘤细胞（U87MG）在体外共同培养,分析其对脑胶质瘤的靶向抗肿瘤作用。结果：hMSCs向胶质瘤细胞的迁徙具有高度特异性。经TRAIL修饰的hMSCs不仅在MSCs表面表达全长TRAIL,且能在培养基中分泌可溶性TRAIL。经rAAV-hTRAIL转染后,hMSCs细胞凋亡作用无增加。将TRAIL修饰的hMSCs与U87MG细胞共培养,与采用可溶性TRAIL治疗后相比较,可明显提高U87MG细胞的凋亡率。结论：在体外实验中,hMSCs向胶质瘤细胞的迁徒具有高度特异性,且能诱导胶质瘤细胞凋亡,并提高对sTRAIL不敏感胶质瘤细胞的凋亡率。由此推测膜结合型TRAIL修饰的MSCs在肿瘤微环境中对脑胶质瘤具备靶向抗肿瘤效应。MSCs可能成为脑胶质瘤靶向治疗的有效载体。     

胶质瘤免疫抑制与治疗的研究进展

正胶质母细胞瘤(glioblastoma multiforme,GBM)具有明显的免疫抑制特性,因此,以恢复胶质瘤相关的免疫作用和促进肿瘤定向免疫的治疗方法在理论上具有可行性,并在啮齿类动物模型和肿瘤免疫治疗的大型临床试验中得到广泛研究。胶质瘤有明显的局部免疫反应,而免疫对中枢神经系统肿瘤的作用及其机制尚不明确。本文讨论胶质瘤的分子和细胞途径介导的免疫系统的抑制作用对胶质瘤免疫治疗     

靶向胶质瘤干细胞及其微环境的免疫治疗进展

胶质瘤是中枢神经系统最常见的原发性恶性肿瘤,胶质瘤干细胞及其微环境是支持胶质瘤发生、恶性进展的根本原因,将其作为包括免疫治疗在内的各项治疗策略的靶点有望提高疗效。单克隆抗体可通过特异性识别胶质瘤干细胞标志物,靶向并抑制其生物学活性;胶质瘤干细胞中肿瘤相关基因变异,使溶瘤病毒选择性复制,裂解胶质瘤干细胞,抑制肿瘤生长;过继转移的嵌合抗原受体T细胞经体外激活、回输后可直接靶向胶质瘤干细胞相关抗原,发挥抗肿瘤作用;树突状细胞疫苗通过胶质瘤干细胞mRNA致敏,发挥靶向激活抗胶质瘤干细胞免疫反应的作用。胶质瘤干细胞活化性配体表达水平升高且低表达主要组织相容性复合物Ⅰ类分子,针对此特点,自然杀伤细胞可以较好地发挥抗肿瘤作用。胶质瘤干细胞微环境中免疫检查点抑制在免疫逃逸过程中起关键作用,对这些免疫检查点进行干预成为肿瘤免疫治疗的重要靶点。靶向胶质瘤干细胞的免疫治疗策略虽在不同程度上显示出一定疗效,但远未成熟,尚待持续深入探究。     

新型弹性蛋白酶抑制剂——西维来司钠的神经保护作用

西维来司钠是新型人工合成的中性粒细胞弹性蛋白酶抑制剂,它在神经保护方面的研究是一个新的领域。研究表明,西维来司钠能够选择性的抑制中性粒细胞弹性蛋白酶(NE)的释放,抑制炎细胞浸润和中性粒细胞活化,减少炎性介质的释放,以各种途径参与颅脑以及脊髓损伤有的神经保护作用,具有良好的临床应用前景。本文将对其生物活性和神经保护作用方面的研究作一综述。     

肿瘤微环境中胶质瘤细胞和免疫细胞代谢对抗肿瘤免疫的调控作用

胶质瘤是中枢神经系统最为常见的恶性肿瘤,呈侵袭性生长,临床难以治愈。近10余年来,手术切除联合同步放化疗和免疫检查点抑制剂的治疗策略取得一定进展,但疗效仍不甚理想,可能与胶质瘤细胞免疫原性较低和肿瘤免疫抑制微环境有关。肿瘤微环境中免疫细胞与胶质瘤细胞及其他激增细胞共同竞争营养物质,竞争过程中产生的代谢产物又可影响免疫细胞分化和功能。本文拟对近年来有关胶质瘤代谢调控与肿瘤免疫抑制微环境的研究进展进行总结,以期为研究肿瘤发生发展机制和胶质瘤治疗提供新的思路。     

间充质干细胞载体基因治疗恶性胶质瘤研究进展

间充质干细胞(mesenchymal stem cells,MSCs)具有肿瘤趋向性生长的能力,并具有抑制肿瘤生长的特性,提示其可能成为治疗恶性胶质瘤的有效载体.MSCs可将特定的细胞因子运至恶性胶质瘤细胞从而发挥抗肿瘤作用,还可通过向恶性胶质瘤分泌抑瘤因子等机制发挥抗肿瘤作用,在恶性胶质瘤的基因治疗中有良好的应用前景.本文就MSCs在恶性胶质瘤基因治疗的研究进展作一综述.     

NLR与脑出血预后关系的研究进展

脑出血具有病残率高和病死率高的特点。炎性反应与脑出血病程密切相关，脑出血后白细胞释放炎性介质与细胞毒性介质，通过提高毛细血管通透性、促进细胞肿胀，损伤血脑屏障，进而增强病变周围水肿，影响脑出血的临床病程。因此，炎症标志物有助于判断脑出血发展过程及预后。在炎症反应中，中性粒细胞和淋巴细胞是两类主要参与者，中性粒细胞与淋巴细胞比值（NLR）则整合了这两类细胞的信息，是炎症反应的可靠指标，该比值在临床应用中存在一定的潜在价值。本文结合相关研究，重点阐述NLR在脑出血后发展过程中的相关作用机制及对预后评估的意义。     

树突状细胞活化的CTLs对神经胶质瘤的体外杀伤作用研究

目的研究负载肿瘤抗原的树突状细胞(DCs)活化的特异性细胞毒性T淋巴细胞(CTLs)对神经胶质瘤细胞的体外杀伤效应,探讨其用于临床治疗的可行性。方法体外原代培养胶质瘤细胞,冻融法获取胶质瘤细胞抗原,联合应用粒细胞/巨噬细胞集落刺激因子、白细胞介素-4和肿瘤坏死因子-α等,对人外周血单核细胞进行体外诱导来获取DCs并负载肿瘤抗原,激活自体T淋巴细胞,制备特异性CTLs,MTT法检测对胶质瘤细胞的体外杀伤效应。结果负载胶质瘤抗原的DCs激活的CTLs对胶质瘤的杀伤作用显著高于对K562细胞的杀伤作用(P<0.01),并且杀伤活性随着效靶比的增加而增加;负载胶质瘤抗原的DCs激活的CTLs对胶质瘤的杀伤活性明显高于未经胶质瘤抗原致敏的DCs刺激的CTLs对胶质瘤的杀伤作用(P<0.01)。结论联合应用细胞因子从人外周血中诱导出的DCs负载胶质瘤抗原后,激活的CTLs在体外对胶质瘤细胞能产生高效而特异的杀伤作用,为临床应用DCs瘤苗奠定基础。     

脑胶质瘤的基因治疗与病毒治疗研究进展

由于脑胶质瘤肿瘤干细胞亚群的耐药性及药物难以通过血-脑屏障和脑组织免疫抑制等特点,目前的常规治疗手段并未明显提升患者的生存时间及生活质量.因此,在分子生物学进步的支持下,对脑胶质瘤新疗法的探索仍在继续.基因治疗包括将治疗用的遗传物质传送到肿瘤细胞中,从而产生特定的抗肿瘤效应.此外,因病毒具有强烈的细胞毒性作用和易于修饰...     

光动力疗法与免疫疗法在肿瘤治疗中的联合应用

近年来．光动力疗法（photodynamic therapy．PDT）在胶质瘤治疗中日渐得到重视．并且随着对中枢神经系统和胶质瘤免疫学特性研究的深入．发现中枢神经系统并非以前认为的“免疫特赦器官”．免疫疗法大有可为。在研究PDT中人们发现，PDT治疗肿瘤时所引发的免疫反应对于控制肿瘤的复发起着至关重要的作用．而这种免疫反应机制复杂，几乎涉及免疫系统的各个方面。因此每一个方面都可能成为一个切人点．可以设计用相应的免疫疗法针对性地与PDT相结合．从而增强PDT激发的抗肿瘤免疫作用．减弱或抑制其引起的免疫抑制反应。     

Regional differences in the Purkinje cells settled pattern: a comparative autoradiographic study in control and homozygous weaver mice

Data accumulated over the last 10 years have led to the popular hypothesis that neutrophils and other inflammatory cells play a prominent role in the neuropathology of cerebral ischemia. This hypothesis was derived from a large number of studies involving three general observations: (1) leukocytes, particularly neutrophils, are present in ischemic tissue at the approximate time that substantial neuronal death occurs; (2) neutropenia is sometimes associated with reduced ischemic damage; and (3) treatments that prevent leukocyte vascular adhesion and extravasation into the brain parenchyma can be neuroprotective. This review reexamines the literature to ascertain its support for a pathogenic role for neutrophils in ischemia-induced neuronal loss. To accomplish this goal, we employed several logical theorems of "cause-effect" relationships, as they pertain to leukocytes and ischemic brain damage. Since the majority of studies focused on neutrophils as the most likely pathogenic inflammatory cell, this review necessarily does so here. We reasoned that if neutrophils play an important pathogenic (i.e., cause-effect) role in the neuronal damage that follows a stroke, then one should expect to find clear evidence that: (1) neutrophils invade the ischemic area prior to terminal stage infarction, (2) greater numbers of early appearing neutrophils are accompanied by evidence of greater neuronal loss, and (3) dose-related inhibition of neutrophil trafficking or activity produces a corresponding decrease in the degree of brain damage following ischemia. This review of the literature reveals that the existing evidence does not readily support any of these predictions and that, therefore, it consistently falls short of establishing a clear cause-effect relationship between leukocyte recruitment and the pathogenesis of ischemia. While the available evidence does not necessarily rule out a potential pathogenic role of neutrophils and other leukocytes, it nevertheless does expose serious weaknesses in the existing studies intended to support that hypothesis. For this reason we also offer suggestions for additional experiments and the inclusion of control groups that, in the future, might provide more effective or conclusive tests of the hypothesis.     

中性粒细胞极性化过程中定向和侧足形成的方法学模式

背景：中性粒细胞极性是其趋化和产生防御功能的基础,而极性化过程中定向能力和侧足形成是重要的环节,目前相关研究的报道还很少。 目的：观察均匀浓度、陡峭浓度梯度、平缓浓度梯度3种实验模式下细胞极性过程中片足的方向和侧足的形成情况,试图找出研究中性粒细胞极性的方法学模式。 方法：①采用密度梯度离心法,成功急性分离人中性粒细胞。②均匀浓度、陡峭浓度梯度和平缓浓度梯度甲酰甲硫氨酰-亮氨酰-苯丙氨酸对细胞进行趋化刺激。③倒置显微镜下观察细胞极性变化情况,判断细胞移动方向。④对细胞进行荧光双标,激光共聚焦显微镜下观察F-actin聚合情况。 结果与结论：①均匀浓度下,中性粒细胞极性化方向和移动轨迹随机分布。②陡峭梯度浓度下,41%的细胞呈现电极向心性分布。③平缓梯度下,68%的细胞表现为沿着甲酰甲硫氨酰-亮氨酰-苯丙氨酸浓度的分布,χ2检验提示平缓浓度梯度实验模式存在较好的定向性。④三者均有明显的侧足形成,以均匀浓度为高,这与定向性呈现为相反的结果。结果显示平缓浓度梯度实验模式在研究细胞定向机制方面相对较好,另一方面对细胞定向的干扰可能来源于细胞侧足的形成。     

Elastase expression by infiltrating neutrophils in gliomas

Abstract

Polymorphonuclear neutrophil elastase is a neutral protease released by activated polymorphonuclear neutrophils and plays a crucial role in maintaining host defense. However, under certain conditions, this enzyme damages normal tissue and facilitates infiltration of tumor cells. In this study, surgical specimens were obtained from the tumor core and infiltrating margin of the glioma of 72 patients with astrocytoma of varying degrees of malignancy, and the specimens were tested for the presence of elastase by immunohistochemical analysis. Polymorphonuclear neutrophil elastase was not present in the tumor core of any of the 12 cases. Elastase was expressed in areas of tumor infiltration of the brain in all four glioblastoma cases, three of the four anaplastic astrocytoma cases, and none of the four low-grade astrocytoma cases. There was a higher percentage of elastase-positive polymorphonuclear neutrophils in the infiltrating margin of tumors with greater degree of malignancy. Polymorphonuclear neutrophils are recruited to malignant gliomas, and the elastase released by these cells aids in the infiltration of gliomas [Neurol Res 2000; 22: 465-468]     

Association of uric acid with traditional inflammatory factors in stroke

Uric acid (UA) plays an important role in the oxidant stress that causes inflammation. We assessed the association between UA and neutrophil ratio, white blood cell (WBC) count and blood lipid in 524 patients admitted with stroke. Stroke patients with a neutrophil ratio >70% displayed significantly lower UA levels than those with a neutrophil ratio ≤70% (p < 0.05). According to UA quartiles, neutrophil ratio, WBC count, and high-density lipoprotein cholesterol in the UA grade 1 group (≤214.10 μmol/L) were significantly increased over those in other UA grade groups. The results of stepwise regression analysis found that UA levels were inversely associated with neutrophil ratios (B ± SE = ?1.11 ± 0.35), high-density lipoprotein cholesterol (B ± SE = ?46.18 ± 14.17), total cholesterol (B ± SE = 9.82 ± 3.66), blood urea nitrogen (B ± SE = 6.30 ± 1.73), and creatinine (B ± SE = 0.63 ± 0.10). There is a correlationship between lower serum uric acid with neutrophil ratios in inflammation associated with stroke and the reasons need to be investigated further.     

Clinical utility of neutrophil–lymphocyte ratio in the diagnosis of neuroleptic malignant syndrome

Abstract

Background: Neuroleptic malignant syndrome (NMS) is a life-threatening side effect of antipsychotic medication. In this study, we aimed to investigate the hypothesis of inflammation via neutrophil–lymphocyte ratio (NLR) in the etiology of NMS.

Methods: In this retrospective case-control study, data were collected using digital database of Bak?rköy Mental Health Research and Training State Hospital by screening NMS diagnosis according to ‘International Classification of Diseases (ICD-10) code: G21.0’ between the years of 2007 and 2017. We included 32 hospitalizations with the diagnosis of NMS and 31 other acute psychiatric hospitalizations without NMS of same patients. NLR was calculated as proportion of absolute neutrophil count to absolute lymphocyte count. Significance level was accepted as p?<?.05.

Results: The mean NLR value of NMS group was 9.55?±?5.13 and control group was 2.06?±?0.71 (p?<?.001). According to ROC analysis in our study group, we found a mean NLR cutoff value ≥4 and lymphocyte percent cutoff of ≤18.4% have the probability of correctly identifying patients with NMS with the 100% sensitivity and 100% specificity.

Conclusions: In this retrospective study, we considered that higher NLR value in NMS episode might be a resemblance of systemic inflammatory state. In addition, our results suggest that both NLR and lymphocyte percentage may be alternative minor criteria which are more sensitive and specific than leukocyte levels and CPK.     

Neutropenia fails to prevent the acute phase stimulation of fibrinogen synthesis

This study evaluates the role of neutrophil granulocytes in mediating acute phase stimulation of fibrinogen synthesis. Turpentine was administered to neutropenic and non-neutropenic rats and fibrinogen synthetic rates measured in an isolated liver perfusion system. Using the (¹⁴C) carbonate technique for the measurement of the absolute synthetic rates of liver produced plasma proteins it was observed that the rates of fibrinogen synthesis of the neutropenic and non-neutropenic rats were significantly greater (p < 0.01) than those of normal control animals, but were not significantly different from each other. These results suggest that the neutrophil granulocyte may not be of major importance in mediating acute phase stimulation of fibrinogen synthesis.     

Distinct pattern of microglial response, cyclooxygenase-2, and inducible nitric oxide synthase expression in the aged rat brain after excitotoxic damage

Microglial and inflammatory responses to acute damage in aging are still poorly understood, although the aged brain responds differently to injury, showing poor lesion outcome. In this study, excitotoxicity was induced by intrastriatal injection of N-methyl-D-aspartate in adult (3-4 months) and aged (22-24 months) rats. Cryostat brain sections were processed for the analysis of microglial response by lectin histochemistry and cyclooxygenase 2 (COX2) and inducible nitric oxide synthase (iNOS) expression by immunohistochemistry and confocal analysis. Aged injured animals showed more widespread area of microglial response at 12 hr postlesion (hpl) and greater microglia/macrophage density at 3 days postlesion (dpl). However, aged reactive microglia showed prevalence of ramified morphologies and fewer amoeboid/round forms. Aged injured animals presented a diminished area of COX2 expression, but a significantly larger density of COX2(+) cells, with higher numbers of COX2(+) neurons during the first 24 hpl and COX2(+) microglia/macrophages later. In contrast, the amount of COX2(+) neutrophils was diminished in the aged. iNOS was more rapidly induced in the aged injured striatum, with higher cell density at 12 hpl, when expression was mainly neuronal. From 1 dpl, both the iNOS(+) area and the density of iNOS(+) cells were reduced in the aged, with lower numbers of iNOS(+) neurons, microglia/macrophages, neutrophils, and astrocytes. In conclusion, excitotoxic damage in aging induces a distinct pattern of microglia/macrophage response and expression of inflammatory enzymes, which may account for the changes in lesion outcome in the aged, and highlight the importance of using aged animals for the study of acute age-related insults.     

大鼠局灶性脑缺血细胞因子介导的中性粒细胞化学吸附剂的动态观察

目的;探索细胞因子介导的中性粒细胞化学吸附剂（CINC）在脑缺血损伤中的作用。方法：用线栓法制备大鼠局灶性脑缺血模型,观察不同脑缺血时程血浆（sera）和脑组织内CINC浓度与髓质过氧化物酶（MPO）活性。结果：缺血组,血浆中CINC浓度的高峰在缺血6小时,脑组织内CINC浓度和髓质过氧化物酶（MPO）活性均呈低水平,其中CINC浓度高峰在缺血12小时,与对照组比较,差异均有极显著性意义（P＜0．01）;血浆中MPO活性随着缺血时间延长而逐渐升高,缺血6小时以上各组与对照组比较差异均有极显著性意义（P＜0．01）;脑组织内各组MPO活性差异无显著性意义（P＜0．05）。结论：缺血时,脑组织内表达的CINC可能是启动缺血性脑损伤的因素之一     

Targeting neutrophils as a novel therapeutic strategy after stroke

Stroke is followed by an intricate immune interaction involving the engagement of multiple immune cells, including neutrophils. As one of the first responders recruited to the brain, the crucial roles of neutrophils in the ischemic brain damage are receiving increasing attention in recent years. Notably, neutrophils are not homogenous, and yet there is still a lack of full knowledge about the extent and impact of neutrophil heterogeneity. The biological understanding of the neutrophil response to both innate and pathological conditions is rapidly evolving as single-cell-RNA sequencing uncovers overall neutrophil profiling across maturation and differentiation contexts. In this review, we scrutinize the latest research that points to the multifaceted role of neutrophils in different conditions and summarize the regulatory signals that may determine neutrophil diversity. In addition, we list several potential targets or therapeutic strategies targeting neutrophils to limit brain damage following ischemic stroke.