D-Glucose but Not Insulin Reduces N-formyl-methionyl-leucyl-phenylalanine (fMet-Leu-Phe)-Induced Shape Changes in Suspended Human Neutrophils

The effects of glucose (5–25 mM) and insulin concentration (40–320 U/ml) on the cell shape of neutrophil granulocytes from healthy humans were studied. Both non-activated and N-formyl-methionyl-leucyl-phenylalanine (fMet-Leu-Phe)-activated neutrophils in suspension were used as a model for initial chemotactic activation of neutrophil locomotion. D-glucose, but not the non-metabolizable analogue 3-O-methyl-D-glucose, dose-dependently reduced the fMet-Leu-Phe-induced (10^–8M) neutrophil elongation. Insulin, either alone or in combination with 25 mM D-glucose, was without effect on the fMet-Leu-Phe-induced neutrophil elongation. Furthermore, the inhibitory effect of D-glucose was observed already after 1 min of exposure to D-glucose and fMet-Leu-Phe. D-glucose diminished the fraction of neutrophils with elongated locomotor shape by changing it into an irregular cell shape, suggesting that at least part of the D-glucose effect could be associated with mechanisms determining the typical locomotor shape. The present results suggest that D-glucose through its metabolism, but without the involvement of insulin, reduces chemotactically induced elongation to a locomotor neutrophil shape, and thus neutrophil motility, and that this effect of glucose appears prior to adhesion. This glucose-induced inhibition of the neutrophil chemotactic response may be involved in the neutrophil deficiency seen in diabetes mellitus.     

The oxidation of apocynin catalyzed by myeloperoxidase: proposal for NADPH oxidase inhibition

Apocynin has been used as an efficient inhibitor of the NADPH oxidase complex and its mechanism of inhibition is linked to prior activation through the action of peroxidases. Here we studied the oxidation of apocynin catalyzed by myeloperoxidase (MPO) and activated neutrophils. We found that apocynin is easily oxidized by MPO/H2O2 or activated neutrophils and has as products dimer and trimer derivatives. Since apocynin impedes the migration of the cytosolic component p47phox to the membrane and this effect could be related to its conjugation with essential thiol groups, we studied the reactivity of apocynin and its MPO-catalyzed oxidation products with glutathione (GSH). We found that apocynin and its oxidation products do not react with GSH. However, this thiol compound was efficiently oxidized by the apocynin radical during the MPO-catalyzed oxidation. We suggest that the reactivity of apocynin radical with thiol compounds could be involved in the inhibitory effect of this methoxy-catechol on NADPH oxidase complex.     

中性粒细胞-淋巴细胞比率与老年NSCLC患者临床病理及预后的相关性分析

目的:研究中性粒细胞-淋巴细胞比率(NLR)与老年非小细胞肺癌(NSCLC)患者临床病理及预后的相关性分析。方法:回顾性分析我院收治的68例老年NSCLC患者的临床病理资料。根据化疗前NLR分为低NLR组(2.95)、高NLR组(≥2.95)。比较两组患者临床病理特点及无病生存期(DFS),并分析预后的影响因素。结果:与低NLR组比较,高NLR组临床分期Ⅳ期、吸烟及淋巴转移的比例更大(P0.05),而两组间年龄、性别、病理类型及合并症比较差异均无统计学意义(P0.05)。低NLR组中位DFS为7.2个月(95%CI:5.9~8.4),显著高于高NLR组中位DFS 6.7个月(95%CI:5.4~7.9)(P0.05)。淋巴转移、NLR是老年NSCLC患者DFS的独立危险因素,而年龄、化疗次数则是独立保护因素(P0.05)。结论:NLR与老年NSCLC患者的临床分期和淋巴结转移有明显相关性,并可作为预后评估参考指标之一。     

红细胞分布宽度血小板比率和中性粒细胞淋巴细胞比率在急诊PCI术后冠脉无复流评估中的应用价值

目的:探讨红细胞分布宽度血小板比率(RDW)和中性粒细胞淋巴细胞比率(NLR)预测急诊经皮冠状动脉介入(PCI)术后冠脉无复流的应用价值。方法:选取2016年1月~2016年12月于我院行急诊PCI手术治疗的患者66例为研究对象,检测其治疗前的RDW和NLR,统计其急诊PCI术后冠脉无复流现象的发生情况,比较冠脉无复流和正常血流患者的RDW和NLR,分析患者RDW和NLR与其冠脉无复流的关系及其对冠脉无复流的预测价值。结果:66例患者急诊PCI术后冠脉无复流发生率为30.30%(20/66)。与正常血流患者比较,冠脉无复流患者RDW和NLR均升高(P0.05)。Spearman相关分析结果显示患者RDW和NLR与其冠脉无复流均呈正相关(r=0.826,0.878,P0.05)。进一步ROC曲线分析结果显示急诊PCI术前RDW和NLR联合预测其术后冠脉无复流的的ROC曲线下面积高于二者单独预测。结论:急诊PCI患者RDW和NLR与其术后冠脉无复流密切相关且两者联合预测患者术后冠脉无复流的价值良好,可能作为急诊PCI术后冠脉无复流评估的参考指标。     

Concomitant elevations of MMP‐9, NGAL,proMMP‐9/NGAL and neutrophil elastase in serum of smokers with chronic obstructive pulmonary disease

A growing body of evidence points towards smoking‐related phenotypic differences in chronic obstructive pulmonary disease (COPD). As COPD is associated with systemic inflammation, we determined whether smoking status is related to serum levels of matrix metalloproteinase‐9 (pro‐ and active MMP‐9), neutrophil gelatinase‐associated lipocalin (NGAL) and the proMMP‐9/NGAL complex in patients with COPD. Serum samples were collected in 100 stable‐phase COPD patients (82 smokers, 18 never‐smokers) and 28 healthy adults (21 smokers, 7 never‐smokers). Serum levels of studied factors were measured in ELISA. Our data provide the first evidence of simultaneously elevated serum levels of MMP‐9, NGAL and proMMP‐9/NGAL in COPD smokers. While the triad discriminated between smokers and non‐smokers in the COPD group, MMP‐9 and proMMP‐9/NGAL (but not NGAL) discriminated between smokers with and without COPD. Adjustment for age and smoking pack‐years did not alter the findings. Serum MMP‐9, NGAL and proMMP‐9/NGAL levels were not correlated with the GOLD stage or FEV1 decline. Furthermore, serum levels of neutrophil elastase (NE) and MMP‐3 (but not of IL‐6 and MMP‐12) were also higher in COPD smokers than in healthy smokers before and after adjustment for age and pack‐years. Among COPD smokers, levels of MMP‐9, NGAL and proMMP‐9/NGAL were positively correlated with NE (P < 0.0001) but not with the remaining factors. Gelatin zymography detected proMMP‐9 in serum samples of healthy and COPD smoking groups. Our results suggest that associated serum levels of proMMP‐9, NGAL, proMMP‐9/NGAL and NE may reflect the state of systemic inflammation in COPD related to cigarette smoking.     

In vivo study of toxoplasmic parasitemia using interferon-gamma-deficient mice: absolute cell number of leukocytes, parasite load and cell susceptibility

Toxoplasma gondii infection was induced in wild type (WT) C57BL/6 and BALB/c mice and same-background interferon-γ knockout (GKO) mice by peroral inoculation of Fukaya strain cysts. We studied parasitemia, absolute cell number of leukocytes, and cell susceptibility in peripheral blood leukocyte (PBL) subsets in vivo. Parasitemia was observed in both WT and GKO mice, although the pattern of the parasite load was totally different among them. After infection, the absolute number of neutrophils in peripheral blood increased in both GKO C57BL/6 and GKO BALB/c mice with statistical significance, while it rose up slightly in susceptible WT C57BL/6 mice, but declined moderately in resistant WT BALB/c mice. The absolute number of lymphocytes in both WT and GKO mice decreased significantly after infection. Although leukocyte number per μl decreased significantly in both strains of WT mice, it increased in GKO BALB/c mice. There were no differences in susceptibility of PBL subsets to T. gondii infection as assessed by quantitative competitive polymerase chain reaction in both WT and GKO mice. These results indicate that each of the PBL subsets was infected in vivo. The increase of neutrophils only among immunocompromised or susceptible strains suggests that the neutrophil may be involved in the lethal process of T. gondii infection. The lack of any difference in cell susceptibility per μg gDNA among the PBL subsets examined implies that the neutrophil may contribute to the whole body dissemination process of the parasite in vivo more than other PBL subsets through the increase in number.     

Fractionation and characterization of biologically-active polysaccharides from Artemisia tripartita

The leaves of Artemisia species have been traditionally used for prevention and treatment of a number of diseases. In this study, five polysaccharide fractions (designated A-I-A-V) were isolated from the leaves of Artemisia tripartita Rydb. by the sequential use of hot-water extraction, ethanol precipitation, ultra-filtration, and chromatography. The homogeneity and average molecular weight of each fraction were determined by high performance size-exclusion chromatography. Sugar composition analysis revealed that Artemisia polysaccharides consisted primarily of xylose, glucose, arabinose, galactose, and galactosamine. Moreover, all fractions contained at least 3.4% sulfate, and fractions A-II-A-V contained an arabinogalactan type II structure. All fractions exhibited macrophage-activating activity, enhancing production of intracellular reactive oxygen species and release of nitric oxide, interleukin 6, interleukin 10, tumor necrosis factor alpha, and monocyte chemotactic protein 1. In addition, all fractions exhibited scavenging activity for reactive oxygen species generated enzymatically or produced extracellularly by human neutrophils. Finally, fractions A-I and A-V exhibited complement-fixing activity. Taken together, our results provide a molecular basis to explain at least part of the beneficial therapeutic effects of Artemisia extracts, and suggest the possibility of using Artemisia polysaccharides as an immunotherapeutic adjuvant.     

Pre-treatment neutrophil to lymphocyte ratio is elevated in epithelial ovarian cancer and predicts survival after treatment

Purpose  Inflammatory cells can both suppress and stimulate tumor growth, and the influence of inflammatory cells on clinical outcome has been the focus of many studies. The purpose of this study was to evaluate the effectiveness of the neutrophil to lymphocyte ratio (NLR), a measure of the systemic inflammatory response, as an additional discriminative biomarker in epithelial ovarian cancer and to determine whether it predicts survival and recurrence. Methods  We studied 192 patients with epithelial ovarian cancer, 173 with benign ovarian tumors, 229 with benign gynecologic disease, and 405 healthy controls. Serum CA125 levels and leukocyte counts according to subtypes were recorded prior to treatment in all study subjects. In epithelial ovarian cancer, the diagnostic usefulness of NLR, in combination with CA125, was evaluated. The correlation between NLR and overall and disease-free survival was analyzed using both univariate and multivariate analyses adjusting for the known prognostic factors (age, stage, cell type, and grade). Results  Preoperative NLR in ovarian cancer subjects (mean 6.02) was significantly higher than that in benign ovarian tumor subjects (mean 2.57), benign gynecologic disease subjects (mean 2.55), and healthy controls (mean 1.98) (P < 0.001). The sensitivity and specificity of NLR in detecting ovarian cancer was 66.1% (95% CI, 59.52–72.68%) and 82.7% (95% CI, 79.02–86.38%), respectively (cutoff value: 2.60). In early stage ovarian cancer, CA125 was not elevated in 19 out of 49 patients. Seven (36.8%) of these 19 patients were NLR positive. On Cox multivariate analysis, NLR positive, stage III/IV, and older age were independent poor prognostic factors, and being NLR positive was the most powerful predictive variable (Hazard Ratio = 8.42 [95% CI: 1.09–64.84], P = 0.041). Conclusions  Our findings provide evidence for the association between NLR and epithelial ovarian cancer. Preoperative NLR, in combination with CA125, may represent a simple and cost-effective method of identifying ovarian cancers, and an elevated NLR may predict an adverse outcome in ovarian cancer.