VEGF signalling enhances lesion burden in KRIT1 deficient mice

The exact molecular mechanisms underlying CCM pathogenesis remain a complicated and controversial topic. Our previous work illustrated an important VEGF signalling loop in KRIT1 depleted endothelial cells. As VEGF is a major mediator of many vascular pathologies, we asked whether the increased VEGF signalling downstream of KRIT1 depletion was involved in CCM formation. Using an inducible KRIT1 endothelial‐specific knockout mouse that models CCM, we show that VEGFR2 activation plays a role in CCM pathogenesis in mice. Inhibition of VEGFR2 using a specific inhibitor, SU5416, significantly decreased the number of lesions formed and slightly lowered the average lesion size. Notably, VEGFR2 inhibition also decreased the appearance of lesion haemorrhage as denoted by the presence of free iron in adjacent tissues. The presence of free iron correlated with increased microvessel permeability in both skeletal muscle and brain, which was completely reversed by SU5416 treatment. Finally, we show that VEGFR2 activation is a common downstream consequence of KRIT1, CCM2 and CCM3 loss of function, though the mechanism by which VEGFR2 activation occurs likely varies. Thus, our study clearly shows that VEGFR2 activation downstream of KRIT1 depletion enhances the severity of CCM formation in mice, and suggests that targeting VEGF signalling may be a potential future therapy for CCM.     

加味星蒌承气汤对急性缺血性脑卒中患者神经功能及血脂、血液流变学的影响

目的:探讨加味星蒌承气汤对急性缺血性脑卒中患者神经功能及血脂、血液流变学的影响。方法:选取2017年8月～2019年6月期间我院收治的急性缺血性脑卒中患者96例,将入选患者根据随机数字表法分为对照组(n=48)和研究组(n=48),对照组患者予以常规西医治疗,研究组患者在对照组基础上联合加味星蒌承气汤治疗,对比两组患者疗效、神经功能及血脂、血液流变学情况,记录两组患者治疗期间不良反应情况。结果:研究组治疗8 d后的临床总有效率为91.67%(44/48),显著高于对照组患者的72.92%(35/48)(P0.05)。两组治疗8 d后加拿大神经功能评分量表(CNS)、美国国立卫生研究所卒中量表(NIHSS)评分均下降(P0.05),且研究组低于对照组(P0.05)。两组治疗8 d后总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、甘油三酯(TG)、全血黏度高切、全血黏度低切、血浆黏度、纤维蛋白原均下降(P0.05),且研究组低于对照组(P0.05);高密度脂蛋白胆固醇(HDL-C)升高(P0.05),且研究组高于对照组(P0.05)。两组不良反应发生率比较无明显差异(P0.05)。结论:急性缺血性脑卒中患者采用加味星蒌承气汤治疗,疗效显著,可有效改善患者神经功能及血脂、血液流变学,且安全性较好。     

Synthesis of Novel Aryloxyethylamine Derivatives and Evaluation of Their in Vitro and in Vivo Neuroprotective Activities

A series of aryloxyethylamine derivatives were designed, synthesized and evaluated for their biological activity. Their structures were confirmed by ¹H‐NMR, ¹³C‐NMR, FT‐IR and HR‐ESI‐MS. The preliminary screening of neuroprotection of compounds in vitro was detected by MTT, and the anti‐ischemic activity in vivo was tested using bilateral common carotid artery occlusion in mice. Most of these compounds showed potential neuroprotective effects against the glutamate‐induced cell death in differentiated rat pheochromocytoma cells (PC12 cells), especially for (4‐fluorophenyl){1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}methanone, {1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}(4‐methoxyphenyl)methanone, (4‐bromophenyl){1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}methanone, {1‐[2‐(4‐chlorophenoxy)ethyl]piperidin‐4‐yl}(4‐chlorophenyl)methanone, (4‐chlorophenyl)(1‐{2‐[(naphthalen‐2‐yl)oxy]ethyl}piperidin‐4‐yl)methanone, (4‐chlorophenyl){1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}methanone and {1‐[2‐(4‐bromophenoxy)ethyl]piperidin‐4‐yl}(4‐chlorophenyl)methanone, which exhibited potent protection of PC12 cells at three doses (0.1, 1.0, 10 μM). Compounds (4‐fluorophenyl){1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}methanone, (4‐fluorophenyl){1‐[2‐(naphthalen‐2‐yloxy)ethyl]piperidin‐4‐yl}methanone, {1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}(4‐methoxyphenyl)methanone and {1‐[2‐(4‐chlorophenoxy)ethyl]piperidin‐4‐yl}(4‐chlorophenyl)methanone possessed the significant prolongation of the survival time of mice subjected to acute cerebral ischemia and decreased the mortality rate at all five doses tested (200, 100, 50, 25, 12.5 mg/kg) and had significant neuroprotective activity. In addition, (4‐fluorophenyl){1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}methanone, {1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}(4‐methoxyphenyl)methanone and {1‐[2‐(4‐chlorophenoxy)ethyl]piperidin‐4‐yl}(4‐chlorophenyl)methanone possessed outstanding neuroprotection in vitro and in vivo. These compounds can be used as a promising neuroprotective agents for future development of new anti‐ischemic stroke agents. Basic structure–activity relationships are also presented.     

阿替普酶联合依达拉奉治疗急性缺血性卒中的疗效及神经功能缺损与时间窗的关系研究

目的:探讨阿替普酶联合依达拉奉治疗急性缺血性卒中的疗效及神经功能缺损与时间窗的关系。方法:选取大连医科大学附属大连市中心医院于2016年3月~2018年10月间收治的急性缺血性卒中患者117例,根据随机数字表法将患者分为对照组(n=58,阿替普酶治疗)和研究组(n=59,阿替普酶联合依达拉奉治疗),比较两组患者临床疗效、神经功能缺损情况、基质金属蛋白酶-9(MMP-9)、白介素-6(IL-6)水平、头颅CT梗死面积,观察两组治疗期间不良反应发生情况。结果:研究组的总有效率为84.75%(50/59),高于对照组的63.79%(37/58)(P<0.05)。两组患者治疗2周后MMP-9、IL-6、美国国立卫生研究院卒中量表(NIHSS)评分均较治疗前降低,且研究组低于对照组(P<0.05)。研究组治疗24h、48h、72h的头颅CT梗死面积小于对照组(P<0.05)。治疗后研究组发病72h内、发病48h内患者NIHSS评分、头颅CT梗死面积高于发病24h内,且发病72h内高于发病48h内(P<0.05)。两组患者不良反应发生率比较无差异(P>0.05)。结论:阿替普酶联合依达拉奉治疗急性缺血性卒中,疗效确切,可有效改善患者过氧化损伤,降低细胞因子水平,且越早的时间窗内接受治疗的患者,其神经功能缺损、脑梗死面积改善效果越好。     

不同病情急性缺血性脑卒中患者血清过氧化还原蛋白1、钙调蛋白、触珠蛋白水平与预后的关系分析

摘要 目的：探讨不同病情急性缺血性脑卒中（AIS）患者血清过氧化还原蛋白1(PRDX1)、钙调蛋白(CAM)、触珠蛋白(HPT)水平与预后的关系。方法：收集2018年4月~2019年4月期间本院收治的127例AIS患者为研究对象,根据患者的病情分为轻度组(39例)、中度组(48例)、重度组(40例),根据患者的改良Rankin 量表( mRS)评分将患者分为预后良好组(73例)和预后不良组(54例),另选同期在我院进行健康检查的健康受试者50例为对照组。对比所有受试者PRDX1、CAM、HPT水平。对比不同预后患者的一般资料、PRDX1、CAM、HPT水平。分析PRDX1、CAM、HPT与NIHSS评分、mRS评分的关系。以多因素Logistic回归分析AIS患者预后的影响因素。结果：轻度组、中度组、中度组的PRDX1、CAM、HPT水平均高于对照组,且随着患者的病情加重,PRDX1、CAM、HPT水平依次升高,差异均有统计学意义(P<0.05)。预后不良组患者的年龄、PRDX1、CAM、HPT均高于预后良好组(P<0.05)。经Pearson检验,AIS患者的PRDX1、CAM、HPT与NIHSS、mRS评分均呈正相关(P<0.05)。经多因素Logistic回归分析可得,年龄较高、PRDX1水平升高、CAM水平升高、HPT水平升高是AIS患者预后不良的危险因素(P<0.05)。结论：AIS患者的PRDX1、CAM、HPT水平异常升高,且与其病情及预后呈现明显的相关性,年龄、PRDX1、CAM、HPT是患者预后的影响因素,对于病情的评估、预后的判断有一定的临床指导价值。     

吡拉西坦联合鼠神经生长因子对急性缺血性脑卒中患者的疗效及Hcy、PCT和皮质醇水平的影响

目的:探讨吡拉西坦联合鼠神经生长因子对急性缺血性脑卒中患者的疗效及对同型半胱氨酸(Hcy)、降钙素原(PCT)和皮质醇水平的影响。方法:回顾性分析我院2017年2月～2018年11月收治的73例急性缺血性脑卒中患者为研究对象,依据入院先后顺序分为对照组(n=35)和观察组(n=38)。对照组患者采用吡拉西坦治疗,观察组基于对照组加以鼠神经生长因子治疗。观察并比较两组临床疗效,治疗前后美国国立卫生研究院卒中量表(NIHSS)、日常生活能力(ADL),治疗前及治疗2周结束时用全自动生化分析仪测定血浆Hcy水平,用放射免疫学分析法测定PCT水平,用化学发光法测定皮质醇水平,用超声多普勒诊断仪测定基底动脉、左右大脑中动脉血流。结果:治疗后,观察组NIHSS评分低于对照组(8.96±1.21)vs(11.27±1.59)分,ADL评分高于对照组(74.21±9.75)vs(66. 04±8.03)分(P0.05)。观察组总有效率高于对照组89.47%vs 68.57%(P0.05)。治疗后,观察组Hcy、PCT及皮质醇水平低于对照组(14.27±2.01)vs (18.51±2.84)μmol/L、(0.25±0.03)vs (0.31±0.04)μg/L、(171.93±23.86)vs(228.75±30.27)nmol/L(P0.05)。治疗后,观察组脑血流学指标高于对照组基底动脉(43.81±6.84)vs(39.62±5.27)mL/min、左大脑中动脉血流(64.27±9.95)vs(57.03±7.52)mL/min、右大脑中动脉血流(62.85±9.01)vs(56.64±7.42)mL/min(P0.05)。结论:吡拉西坦联合鼠神经生长因子能够提高急性缺血性脑卒中的疗效,降低Hcy、PCT及皮质醇水平,促进神经功能的恢复。     

Electrofishing‐induced cardiac disturbance and injury in rainbow trout

Cardiac output (CO), heart rate (HR) and stroke volume (SV) were monitored in rainbow trout Oncorhynchus mykiss electroshocked (pulsed DC) with various voltage, frequency, pulse width and shock duration settings. Additionally, behavioural recovery times and internal haemorrhaging were examined. During electroshock, heart function became erratic and typically ceased for much of the event (cardiac arrest). Following electroshock, CO increased almost entirely due to an increase in SV. Cardiac function generally returned to resting levels within 2–3 h. Behaviourally, recovery was much more rapid, typically taking only a few minutes for fish to regain equilibrium and to begin swimming normally, and only rarely was >1 h. Internal injury ranged from 0 to 7 cm² of haemorrhaging along the spine and surrounding musculature, although only 4% of the fish had corresponding damage to the vertebrae. Comparisons across the various electrofisher settings indicate that response and injury are highly variable, but generally increased with more intense settings. Higher frequencies and voltages appear to most negatively affect behavioural recovery and injury while longer shock durations increased the length of cardiac arrest and the duration of cardiac recovery. Because of the variability in results and small sample sizes, however, the strongest conclusion that can be drawn from this work is that electrofishing, regardless of the settings, has a considerable negative impact on rainbow trout that is often not apparent externally. Additionally, this study has shown how electrofishing directly effects fish physiologically through impairment of cardiac function. Studies examining the physiological and behavioural response of fishes and subsequent recovery will be necessary for the development of electrofishing guidelines which minimize the disturbance to fishes.     

The combination of mannitol and temozolomide increases the effectiveness of stem cell treatment in a chronic stroke model

Background

The blood-brain barrier (BBB) presents a significant challenge to the therapeutic efficacy of stem cells in chronic stroke. Various methods have been developed to increase BBB permeability, but these are associated with adverse effects and are, therefore, not clinically applicable. We recently identified that combination drug treatment of mannitol and temozolomide improved BBB permeability in vitro. Here, we investigated whether this combination could increase the effectiveness of stem cell treatment in an animal model of chronic ischemic stroke.

Hemopexin increases the neurotoxicity of hemoglobin when haptoglobin is absent

Hemopexin (Hpx) binds heme with extraordinary affinity, and after haptoglobin may provide a second line of defense against the toxicity of extracellular hemoglobin (Hb). In this series of experiments, the hypothesis that Hpx protects neurons from Hb neurotoxicity was evaluated in murine primary cultures containing neurons and glial cells. Contrary to hypothesis, Hpx increased neuronal loss due to micromolar concentrations of Hb by 4‐ to 12‐fold, as measured by LDH release assay; conversely, the neurotoxicity of hemin was completely prevented. The endogenous fluorescence of Hpx was quenched by Hb, consistent with transfer of Hb‐bound heme to Hpx. This was associated with precipitation of globin chains, as detected by immunostaining and fluorescent Hb labeling. A portion of this precipitate attached firmly to cells and could not be removed by multiple washes. Concomitant treatment with haptoglobin (Hp) prevented globin precipitation and most of the increase in neuronal loss. Hpx weakly attenuated the increase in culture non‐heme iron produced by Hb treatment, quantified by ferrozine assay. However, Hb‐Hpx toxicity was iron‐dependent, and was blocked by deferoxamine and ferrostatin‐1. Up‐regulation of cell ferritin expression, a primary cell defense against Hb toxicity, was not observed on western blots of culture lysates that had been concomitantly treated with Hpx. These results suggest that Hpx destabilizes Hb in the absence of haptoglobin, leading to globin precipitation and exacerbation of iron‐dependent oxidative cell injury. Combined therapy with hemopexin plus haptoglobin may be preferable to hemopexin alone after CNS hemorrhage.