男性高血压患者血尿酸与勃起功能障碍及血管内皮损伤的关系#br#

摘要：目的　探讨男性高血压患者血清尿酸（UA）水平变化及其与勃起功能障碍（ED）和血管内皮损伤指标一氧化氮（NO）、内皮型一氧化氮合酶（eNOS）和内皮微粒（EMP）的关系。方法　选择新诊断男性高血压患者为研究对象,采用ED国际指数问卷表-5（IIEF-5）评估ED及其严重程度,100例伴ED患者（轻度31例、中度34例、重度35例）为ED组,100例年龄配对无ED的患者为对照组,尿酸酶法测定血清UA,酶联免疫法测定NO、eNOS,流式细胞技术测定血浆EMP水平。结果　重度ED组UA和EMP高于其他各组;而NO和eNOS低于其他各组,中度ED组UA和EMP高于轻度组及对照组;而NO和eNOS低于轻度组及对照组,轻度组UA和EMP高于对照组;而NO和eNOS低于对照组（P＜0.05）。相关分析表明高血压患者UA与NO、eNOS呈负相关,与EMP呈正相关（r分别为-0.589、-0.693和0.717,均P＜0.01）。多元逐步回归分析结果显示高血压患者UA、低密度脂蛋白胆固醇（LDL-C）、收缩压、糖尿病为EMP的影响因素,多元Logistic回归分析显示UA、舒张压、BMI及LDL-C为高血压患者ED的独立影响因素（P＜0.01）,血清UA预测高血压伴ED的ROC曲线下面积为0.785（95%CI：0.717～0.852,P＜0.01）。以血清UA 384.5 µmol/L作为预测ED最佳阈值时,敏感度90%,特异度69%,阳性预测值74.8%,阴性预测值87.3%。结论　高血压合并ED患者血清UA水平增高,高尿酸血症可能通过损伤血管内皮功能促使高血压患者ED发生与发展。     

Cytotoxicity,cellular uptake and apoptotic responses in human coronary artery endothelial cells exposed to ultrasmall superparamagnetic iron oxide nanoparticles

Ultrasmall superparamagnetic iron oxide nanoparticles (USPION) possess reactive surfaces, are metabolized and exhibit unique magnetic properties. These properties are desirable for designing novel theranostic biomedical products; however, toxicity mechanisms of USPION are not completely elucidated. The goal of this study was to investigate cell interactions (uptake and cytotoxicity) of USPION using human coronary artery endothelial cells as a vascular cell model. Polyvinylpirrolidone-coated USPION were characterized: average diameter 17 nm (transmission electron microscopy [TEM]), average hydrodynamic diameter 44 nm (dynamic light scattering) and zeta potential −38.75 mV. Cells were exposed to 0 (control), 25, 50, 100 or 200 μg/mL USPION. Concentration- and time-dependent cytotoxicity were observed after 3-6 hours through 24 hours of exposure using Alamar Blue and Real-Time Cell Electronic Sensing assays. Cell uptake was evaluated by imaging using live-dead confocal microscopy, actin and nuclear fluorescent staining, and TEM. Phase-contrast, confocal microscopy, and TEM imaging showed significant USPION internalization as early as 3 hours after exposure to 25 μg/mL. TEM imaging demonstrated particle internalization in secondary lysosomes with perinuclear localization. Three orthogonal assays were conducted to assess apoptosis. TUNEL staining demonstrated a marked increase in fragmented DNA, a response pathognomonic of apoptosis, after a 4-hour exposure. Cells subjected to agarose gel electrophoresis exhibited degraded DNA 3 hours after exposure. Caspase-3/7 activity increased after a 3-hour exposure. USPION uptake resulted in cytotoxicity involving apoptosis and these results contribute to further mechanistic understanding of the USPION toxicity in vitro in cardiovascular endothelial cells.     

Liraglutide improves obesity-induced renal injury by alleviating uncoupling of the glomerular VEGF–NO axis in obese mice

Obesity-related kidney disease is associated with generalized endothelial dysfunction. Liraglutide, a glucagon-like peptide-1 agonist, has cardiovascular–renal protective effects in patients with diabetes. In this study, the ability of liraglutide to reduce urinary albumin excretion by alleviating glomerular vascular endothelial growth factor-nitric oxide (VEGF–NO) axis uncoupling was assessed in high fat diet-induced obese mice. C57BL/6J mice were divided into control and obesity groups, treated with or without liraglutide (200 μg/kg/day). Blood biochemistry and urinary albumin excretion were measured. Glomerular VEGF and the AMPK–endothelial nitric oxide synthase (eNOS) pathway were assayed by western blotting. Glomerular NO, renal haeme oxygenase-1 activity, and malondialdehyde levels were also measured. Treatment of obese mice with liraglutide led to significant reductions in body weight gain (46 ± 1 g vs 55 ± 1 g, P < .0001), visceral fat (8.9 ± 0.6 g vs 14.5 ± 0.6 g, P < .0001), perirenal fat (2.9 ± 0.2 g vs 5.4 ± 0.3 g, P < .0001), and free fatty acid (1.71 ± 0.12 mmol/L vs 1.02 ± 0.08 mmol/L, P < .0001). Liraglutide significantly improved glucose homeostasis, which was impaired in obese mice. Liraglutide reduced urinary albumin excretion and glomerular hypertrophy in obese mice. Additionally, liraglutide significantly decreased VEGF and increased glomerular NO production in glomeruli, indicating restoration of the glomerular VEGF–NO axis. Furthermore, liraglutide activated the glomerular AMPK–eNOS pathway in obese mice, upregulated renal haeme oxygenase-1 activity, and reduced the renal malondialdehyde levels in obese mice. In conclusion, liraglutide reduced microalbuminuria and ameliorated renal injury by alleviating the uncoupling of the glomerular VEGF–NO axis.     

Iron-Oxide Nanocluster Labeling of Clostridium novyi-NT Spores for MR Imaging–Monitored Locoregional Delivery to Liver Tumors in Rat and Rabbit Models

PurposeTo label Clostridium novyi-NT spores (C. novyi-NT) with iron oxide nanoclusters and track distribution of bacteria during magnetic resonance (MR) imaging-monitored locoregional delivery to liver tumors using intratumoral injection or intra-arterial transcatheter infusion.Materials and MethodsVegetative state C. novyi-NT were labeled with iron oxide particles followed by induction of sporulation. Labeling was confirmed with fluorescence microscopy and transmission electron microscopy (TEM). T2 and T2* relaxation times for magnetic clusters and magnetic microspheres were determined using 7T and 1.5T MR imaging scanners. In vitro assays compared labeled bacteria viability and oncolytic potential to unlabeled controls. Labeled spores were either directly injected into N1-S1 rodent liver tumors (n = 24) or selectively infused via the hepatic artery in rabbits with VX2 liver tumors (n = 3). Hematoxylin-eosin, Prussian blue, and gram staining were performed. Statistical comparison methods included paired t-test and ANOVA.ResultsBoth fluorescence microscopy and TEM studies confirmed presence of iron oxide labels within the bacterial spores. Phantom studies demonstrated that the synthesized nanoclusters produce R2 relaxivities comparable to clinical agents. Labeling had no significant impact on overall growth or oncolytic properties (P >.05). Tumor signal-to-noise ratio (SNR) decreased significantly following intratumoral injection and intra-arterial infusion of labeled spores (P <.05). Prussian blue and gram staining confirmed spore delivery.ConclusionsC. novyi-NT spores can be internally labeled with iron oxide nanoparticles to visualize distribution with MR imaging during locoregional bacteriolytic therapy involving direct injection or intra-arterial transcatheter infusion.     

Are fluorescence-based chlorophyll quantification methods suitable for algae toxicity assessment of carbon nanomaterials?

Using a multiwalled carbon nanotube (MWCNT) and graphene oxide (GO) as representative test materials, we evaluated the applicability of in vivo and in vitro chlorophyll-a (Chl-a) fluorescence quantification methods, which are used in standard algae ecotoxicity tests such as OECD 201 and ISO 8692. In vivo quantification of Chl-a from Raphidocelis subcapitata indicated a significant reduction in Chl-a fluorescence in the presence of MWCNTs due to shading, but a significant autofluorescence from GO caused an overestimation of Chl-a concentration. In vitro Chl-a quantification methods employing a modified acetone and an ethanol extraction protocol reduced the influence of shading and autofluorescence, but both resulted in a significant loss of fluorescence signal in the presence of 100?mgL^?1 MWCNTs (99–100%) and GO (21–52%). Chl-a reduction was dose dependent for both tested carbon-based MNMs (CNMs), but effects were more pronounced for MWCNT, which caused a significant fluorescence reduction (16?±?0.3%) already at 1?mgL^?1. Further study of the CNM–algae–Chl-a interaction processes revealed that CNM can not only interact with live algae, but also efficiently adsorb extracted Chl-a. Our results showed that within 10?min, 95–100% of Chl-a extracted from two algae concentrations were adsorbed to MWCNT, while 35–60% of Chl-a was adsorbed to the GO. This study shows that Chl-a quantification by fluorescence determination is not a suitable method for ecotoxicity testing of CNM. However, a quick screening test for individual MNMs is recommended to determine whether Chl-a adsorption is a significant process prior to selection of a quantification method.     

The elemental changes occurring in the rat liver after exposure to PEG-coated iron oxide nanoparticles: total reflection x-ray fluorescence (TXRF) spectroscopy study

The main goal of this study was to evaluate in vivo effects of low dose of PEG-coated magnetic iron oxide nanoparticles (IONPs) on the rat liver. The IONPs was intravenously injected into rats at a dose equaled to 0.03?mg of Fe per 1?kg of an animal body weight. The elemental composition of liver tissue in rats subjected to IONPs action and controls were compared. Moreover, in order to determine the dynamics of nanoparticles (NPs) induced elemental changes, the tissues taken from animals 2?hours, 24?hours, and 7?days from IONPs injection were examined. The analysis of subtle elemental anomalies occurring as a result of IONPs action required application of highly sensitive analytical method. The total reflection X-ray fluorescence spectroscopy perfectly meets such requirements and therefore it was used in this study. The obtained results showed increasing trend of Fe level within liver occurring 2?hours from IONPs injection. One day after NPs administration, the liver Fe content presented the baseline level what suggests only the short-term accumulation of nanoparticles in the organ. The Ca, Cu, and Zn levels changed significantly as a result of NPs action. Moreover, the anomalies in their accumulation were still observed 7?days after IONPs injection. The level of Cu decreased while those of Ca and Zn increased in the liver of NPs-treated animals. The reduced liver Cu, followed by elevated serum level of this element, might be related in triggering the mechanisms responsible for Fe metabolism in the organism.     

气相色谱法测定药用辅料聚西托醇1000中的残留杂质

建立气相色谱法测定聚西托醇1000中残留的环氧乙烷、1,4-二氧六环、乙二醇、二甘醇和三甘醇等杂质,为聚西托醇1000生产质量控制提供参考。采用DB-1色谱柱检测环氧乙烷和1,4-二氧六环,顶空进样,进样口温度150℃,检测器温度250℃,顶空平衡温度70℃,平衡时间45 min。采用VF-17MS色谱柱检测乙二醇、二甘醇和三甘醇,液体进样,进样口温度270℃,检测器温度290℃。实验结果显示,环氧乙烷和1,4-二氧六环在各加样量范围内线性良好（r> 0.999）,精密度RSD小于8.0%,平均回收率分别为90.6%和101.2%;乙二醇、二甘醇和三甘醇在3～60μg/mL内线性关系良好（r>0.999）,精密度RSD小于3.0%,回收率均在96%～103%。本研究所建立的方法具有良好的专属性、线性、精密度和回收率,能够有效检测聚西托醇1000中多组分极微量杂质。     

呼出气一氧化氮测定在COPD频繁急性加重表型患者中的临床意义

目的 探讨中央和外周气道呼出气一氧化氮(FeNO)在慢性阻塞性肺病(COPD)频繁急性加重表型患者中的临床意义。方法 收集2019年1月至2021年6月安徽医科大学第四附属医院呼吸与危重症医学科收治的90例COPD患者作为研究对象,在一年内因急性加重次数≥2次的患者纳入频繁急性加重组(n=23),急性加重次数≤1次的患者纳入非频繁急性加重组(n=67)。测定两组中央气道一氧化氮(FeNO₅₀)浓度和外周气道呼出气一氧化氮(FeNO₂₀₀)浓度、肺泡气一氧化氮(CaNO)浓度并收集白细胞计数、血嗜酸粒细胞百分比、血清总免疫球蛋白E(IgE)、慢性阻塞性肺疾病评估测试(CAT)评分及肺功能等相关指标,对进行对比分析。结果 COPD频繁急性加重组FeNO₂₀₀、CaNO、C反应蛋白(CRP)、血清总IgE、CAT评分均高于非频繁急性加重组,第一秒用力呼气容积占预计值百分比(FEV1%)低于非频繁急性加重组(P＜0.05)。在COPD急性加重患者中,FeNO₂₀₀、CaNO与FEV1%呈负相关(r=-0.305、-0.439,P＜0.05),与血清总IgE呈正相关(r=0.523、0.514,P＜0.05),其中CaNO还与CRP呈正相关(r=0.321,P=0.023)。结论 COPD频繁急性加重表型患者中央和外周FeNO浓度存在差异,FeNO₂₀₀、CaNO均可作为COPD患者外周小气道炎症指标。