scholarly journals Isolation and identification of serum exosomes and correlation of its contents with acute ischemic stroke

Author(s):  
Pei Yu ◽  
Wencheng Chen

Abstract Background Erythrocyte deformability is one of the pathophysiological changes of high-risk factors such as smoking, hypertension and atherosclerosis in stroke. It mainly affects blood viscosity and fluidity in the occurrence, development and outcome of the disease. Exosomes are a new type of biological activity test target, and it mainly contents miRNA, but its effect on stroke is still not clear. Objective To detect the serum exosome-derived miR-150-5p expression in patients with acute ischemic stroke and explore its diagnostic potential for acute ischemic stroke. Methods A total of 84 samples were collected with matched age and gender, collecting relevant laboratory indicators and general clinical data of the research subjects. The kit was used to extract serum exosomes, real-time fluorescent quantitative polymerase chain reaction was used to determine the expression of serum exosomal miR-150-5p, evaluate its value as a diagnostic marker through the ROC curve. Through Randa, Target Scan and other online databases, bioinformatics methods predict the target genes of miR-150, the results are drawn Venny to take intersection analysis, literature screening and ischemic stroke related target genes. Results The exosomes showed elliptical or round membranous vesicles with a diameter between 30–200 nm and fusion phenomenon and detected the exosomal marker proteins CD63 and HSP70; Compared with the control group, the relative expression of exosomal miR-150-5p in patients with acute ischemic stroke was increased (T = 8, P < 0.001). The expression of serum exosomal miR-150-5p in the disease group at different time points after stroke, the difference was not statistically significant(Chi-square = 2.925, P = 0.232); the area under the ROC curve was 0.883. bioinformatics prediction analysis of miR-150 target genes, which may be involved in the development of stroke disease through EGR2 and PLP2, as well as erythrocyte membrane protein-related genes SLC4A1 and SPTB also belong to the miR-150 prediction targets. Conclusion The expression of exosome-derived miR-150-5p is relatively high in patients with acute stroke, and it has a certain potential for early disease diagnosis.

2019 ◽  
Vol 20 (10) ◽  
pp. 781-784 ◽  
Author(s):  
Meizhen Zhao ◽  
Li Juanjuan ◽  
Fan Weijia ◽  
Xie Jing ◽  
Huang Qiuhua ◽  
...  

Background: This study aimed to investigate the expression levels of microRNA (miRNA)-125b in serum exosomes and its diagnostic efficacy for asthma severity. Methods: The study included 80 patients with untreated asthma and 80 healthy volunteers. The patients were divided into 4 groups according to disease severity: 20 with the intermittent state, 20 with the mildly persistent state, 20 with the moderately persistent state, and 20 with the severely persistent state. The expression levels of miRNA-125b in serum exosomes of each group were detected using a quantitative polymerase chain reaction and compared. The Spearman correlation analysis was used to study the correlation between the expression levels of miRNA-125b in serum exosomes and asthma severity. The diagnostic efficacy of the expression levels of miRNA-125b in exosomes for asthma severity was evaluated using the Receiver Operating Characteristic (ROC) curve. Results: The expression levels of miRNA-125b in serum exosomes of patients with intermittent, mildly persistent, moderately persistent, and severely persistent asthma were all higher than those in the healthy control group, with statistically significant differences. The expression levels of miRNA-125b were also statistically significantly different among patients in each group. The Spearman correlation analysis showed a positive correlation of the relative expression of miRNA-125b in serum exosomes with asthma severity. The area under the ROC curve of the diagnostic efficacy of miRNA-125b in serum exosomes for patients with intermittent, mildly, moderately, and severely persistent asthma was 0.7770, 0.8573, 0.9111, and 0.9995, respectively. Conclusion: The expression levels of miRNA-125b in serum exosomes had a high diagnostic efficacy and might serve as a noninvasive diagnostic marker for asthma severity.


2021 ◽  
pp. 0271678X2110249
Author(s):  
Giorgio FM Cattaneo ◽  
Andrea M Herrmann ◽  
Sebastian A Eiden ◽  
Manuela Wieser ◽  
Elias Kellner ◽  
...  

Selective therapeutic hypothermia (TH) showed promising preclinical results as a neuroprotective strategy in acute ischemic stroke. We aimed to assess safety and feasibility of an intracarotid cooling catheter conceived for fast and selective brain cooling during endovascular thrombectomy in an ovine stroke model. Transient middle cerebral artery occlusion (MCAO, 3 h) was performed in 20 sheep. In the hypothermia group (n = 10), selective TH was initiated 20 minutes before recanalization, and was maintained for another 3 h. In the normothermia control group (n = 10), a standard 8 French catheter was used instead. Primary endpoints were intranasal cooling performance (feasibility) plus vessel patency assessed by digital subtraction angiography and carotid artery wall integrity (histopathology, both safety). Secondary endpoints were neurological outcome and infarct volumes. Computed tomography perfusion demonstrated MCA territory hypoperfusion during MCAO in both groups. Intranasal temperature decreased by 1.1 °C/3.1 °C after 10/60 minutes in the TH group and 0.3 °C/0.4 °C in the normothermia group (p < 0.001). Carotid artery and branching vessel patency as well as carotid wall integrity was indifferent between groups. Infarct volumes (p = 0.74) and neurological outcome (p = 0.82) were similar in both groups. Selective TH was feasible and safe. However, a larger number of subjects might be required to demonstrate efficacy.


2020 ◽  
Author(s):  
Jun Ni ◽  
Huisheng Chen ◽  
Guofang Chen ◽  
Yong Ji ◽  
Fei Yi ◽  
...  

Abstract Background: Ischemic stroke is a leading cause of morbidity and mortality. Thrombolytic therapy improves disability and survival rates; however, to be effective, it must be given within 4.5 hours of onset. Moreover, thrombolytic therapy is frequently contraindicated. Therefore, alternative therapeutic options are required. In China, cinepazide maleate injection has been shown to improve the cerebral collateral circulation and further reduce disability in stroke patients; however, very few studies investigating this therapy have been conducted to date. Therefore, this study aimed to further confirm the efficacy and safety of cinepazide maleate injection in patients with acute ischemic stroke.Methods: Patients with acute ischemic stroke were administered an intravenous infusion of 320 mg cinepazide maleate or placebo once daily for 14 days. All patients were also administered basic therapy (citicoline sodium). The primary efficacy endpoint was the proportion of patients with a modified Rankin scale (mRS) ≤2 on day 90. Secondary efficacy endpoints included Barthel Index ≥95. Safety was evaluated by recording all adverse events (AEs), monitoring laboratory parameters and vital signs, and electrocardiogram.Results: In total, 937 patients with an acute ischemic stroke were included, with a mean (standard deviation, SD) National Institutes of Health Stroke Scale score of 8.8 (2.4) and a mean (SD) stroke onset of 30.9 (11.4) hours prior. Following treatment for 90 days, the proportion of patients with an mRS score ≤2 was significantly higher in the cinepazide maleate group than in the control group (60.9% vs. 50.1%; p=0.0004). Moreover, the proportion of patients with a Barthel Index of ≥95 on day 90 was also significantly higher in the cinepazide maleate group than in the control group (53.4% vs. 46.7%; p=0.0230). There were no statistically significant differences in safety parameters between the cinepazide maleate and control groups.Conclusions: The results of this study show that cinepazide maleate injection is superior to placebo in improving neurological function and activities of daily living, reducing disability, and promoting functional recovery in patients with acute ischemic stroke. Cinepazide maleate injection was safe and well tolerated with no unexpected AEs reported.Trial registration: Chinese Clinical Trial Registry CTR20160292 and ChiCTR1900023827. Retrospectively registered June 13, 2019.


2020 ◽  
Author(s):  
Jung-Won Choi ◽  
In Woo Ryoo ◽  
Jun Yeong Hong ◽  
Kyung-Yul Lee ◽  
Hyo Suk Nam ◽  
...  

Abstract Background: Sex hormones may be associated with a higher incidence of ischemic stroke or stroke-related events. In observational studies, lower testosterone concentrations are associated with infirmity, vascular disease, and adverse cardiovascular risk factors. Currently, female sexual hormones are considered neuroprotective agents. The purpose of this study was to assess the role of sex hormones and the ratio of estradiol/testosterone (E/T) in patients with acute ischemic stroke (AIS).Methods: Between January 2011 and December 2016, 146 male patients with AIS and 152 age- and sex-matched control subjects were included in this study. Sex hormones, including estradiol, progesterone, and testosterone, were evaluated in the AIS patient and control groups. We analyzed the clinical and physiological levels of sex hormones and hormone ratios in these patients.Results: The E/T ratio was significantly elevated among patients in the stroke group compared to those in the control group (P = 0.001). Categorization of data into tertiles revealed that patients with the highest E/T ratio were more likely to have AIS [odds ratio (OR) 3.084; 95% Confidence interval (CI): 1.616-5.886; P < 0.001) compared with those in the first tertile. The E/T ratio was also an independent unfavorable outcome predictor with an adjusted OR of 1.167 (95% CI: 1.053-1.294; P = 0.003).Conclusions: These findings support the hypothesis that increased estradiol and reduced testosterone levels are associated with AIS in men.


Stroke ◽  
2021 ◽  
Vol 52 (Suppl_1) ◽  
Author(s):  
Lacy S Handshoe ◽  
Joshua Santucci ◽  
Takashi Shimoyama ◽  
Ken Uchino

Background: Non-occlusive thrombus in an intracranial artery in acute ischemic stroke is an uncommon occurrence. We compared the clinical course and outcome of intracranial subocclusive to occlusive thrombi. Methods: We conducted a review of patients who presented with acute ischemic stroke and received CT angiogram at a single comprehensive stroke center from January 2018 to December 2019. Patients with intracranial subocclusive thrombus were compared to a control group with complete occlusion matched for occlusion location. Subocclusive thrombus was reviewed by two raters on CT angiography, disagreement resolved by consensus. Patient and stroke characteristics and the clinical course were analyzed. Neurological deterioration was defined as an increase in NIH Stroke Scale (NIHSS) score > 4 compared from baseline to 48 hours. Good outcome at discharge was defined as modified Rankin Score of ≤2. Results: Among 1151 acute ischemic strokes, there were 896 patients with CT angiograms. Sixteen out of 896 (1.8%) patients had intracranial subocclusive thrombus. Thirty-two with comparable intracranial occlusions were identified. In the subocclusive group, 3 of 16 (19%) of received acute endovascular intervention, compared to 13 of 32 (41%) in the occluded group. Sex, median age or time from last known well to hospital arrival did not differ between the two groups. The subocclusive thrombus group had less severe strokes, with median NIHSS score at arrival 3 compared to 8.5 in the occlusion group (p<0.01) and median NIHSS at discharge 1 compared to 5.5 in the occlusive group (p<0.01). Frequency of neurological deterioration in hospital did not differ between the subocclusive and occluded groups at 48 hours (15% vs 15% p=1.00). The subocclusive group was associated with a good outcome at discharge, OR 0.5.71, 95% confidence interval 1.41-23.1. Conclusion: Intracranial subocclusive thrombus in acute ischemic stroke has a more mild presentation compared to complete intracranial occlusion without a high rate of neurological deterioration.


Stroke ◽  
2017 ◽  
Vol 48 (suppl_1) ◽  
Author(s):  
Jeffrey Leya ◽  
Elisabeth Donahey ◽  
Megan Rech

Introduction: Early treatment of acute ischemic stroke (AIS) with recombinant tissue plasminogen activator (rtPA) within 4.5 hours of symptom onset is associated with neurologic improvement. A risk of rtPA is hemorrhagic conversion, which has a higher incidence in patients with elevated blood pressure at presentation. Current literature supports the use of blood pressure goals (<185/110 mm Hg) in patients qualifying for rtPA, but the effects of anti-hypertensive (anti-HTN) medications within the first 24 hours of AIS on outcomes has not been evaluated. Hypothesis: AIS patients requiring anti-HTN medications (anti-HTN group) before rtPA have a poorer outcome at 90 days compared to those that do not need anti-HTN medications (control group). Methods: This was a retrospective cohort study of patients >18 years diagnosed with AIS from January 2011 through December 2015 who received one or multiple anti-HTN medication(s) prior to rtPA administration, compared to control patients who did not. Primary endpoint was poor outcome at 90 days, defined as a modified Rankin Scale (mRS) of ≥3. Univariate analysis with Chi-square, Fisher’s exact test or t-test was performed. Multivariate analysis was conducted. Results: Of 235 patients evaluated for AIS, 145 (61.7%) were included. Baseline demographics were well matched, though more patients in the anti-HTN group had a history of HTN (86.7% vs. 62.5%, p<0.01), diabetes (33.3% vs. 17.5%, p=0.04) and chronic kidney disease (20% vs. 7.5%, p=0.04). There was no difference in the primary endpoint of poor outcome (mRS ≥3) between groups who received blood pressure medication versus those who did not (37% anti-HTN group vs. 30% control, p=.374). There was no difference in hemorrhagic conversion (13.3% anti-HTN group vs. 6.3% control, p=.187). Mortality at 90 days did not differ between groups (11% who received anti-HTN vs. 7.5%, p=.508). Conclusion: No difference was observed in poor outcomes, hemorrhagic conversion, or 90-day mortality in patients receiving anti-HTN medications prior to rtPA compared to those that did not. These results suggest that aggressive blood pressure management should be used to control hypertension in AIS who may qualify for rtPA, though larger, randomized trials are needed to confirm this finding.


Stroke ◽  
2017 ◽  
Vol 48 (suppl_1) ◽  
Author(s):  
Jon Schrock ◽  
Ashley Markowski

Background: Identification of acute ischemic stroke patients (AIS) with large vessel occlusion (LVO) in the pre-hospital setting is paramount now that endovascular treatment has become the standard of care. Correctly identifying this AIS population will allow transport to appropriate stroke centers with endovascular stroke capabilities. Objectives: We choose to evaluate single symptoms which, if present, might signify an ischemic stroke with a LVO. We tested if the following stroke symptoms including hemiplegia, severe dysarthria, neglect, eye deviation were associated with a LVO. Our hypothesis is that the use of these selected single or multiple NIHSS symptoms will not result in a ROC of > 0.9 rendering it effective in LVO diagnosis. Methods: A retrospective cohort review was conducted of 551 AIS patients who presented to the emergency department over a 77 month period. Subject demographics clinical symptoms, and other pertinent information were collected from the medical record. The following symptoms at ED presentation were evaluated with univariate and multivariate regression and ROC curve analysis; hemiplegia (of arm and leg), dysarthria (severe), neglect, and eye deviation. Results: Out of the 551 patients, 220 (40%) had a moderate stroke (NIHSS >9) while 143 (26%) had a severe stroke (NIHSS>14) and 49% were male. The median NIHSS was 7 (IQR 4-15) and 228 (41%) had LVO on CTA. The odds ratio and area under the ROC curve for hemiplegia, severe dysarthria, neglect, and eye deviation were respectively OR 3.2 (2.2- 4.5), 3.2 (2.2- 4.6), 5.5 (3.4- 8.9), 7.7 (4.6- 13.1) and area under the ROC 0.63, 0.64, 0.63, and 0.64 respectively. The results of multivariate analysis were slightly better with an ROC of 0.72. Conclusions: Singular stroke symptoms do not have the diagnostic power to predict which patient with AIS will have a LVO. While they have positive odds ratios, the ROC analysis suggests that individually or in combination, they are not accurate enough to determine the presence of LVO.


Stroke ◽  
2017 ◽  
Vol 48 (suppl_1) ◽  
Author(s):  
Saif Bushnaq ◽  
Atif Zafar ◽  
Kempuraj Duraisamy ◽  
Nudrat Tasneem ◽  
Mohammad M Khan ◽  
...  

Background: Interleukin-37 (IL-37) is a new member of IL-1 cytokine family with a defined role as a negative feedback inhibitor of pro-inflammatory responses. IL-37 has yet to be evaluated in non-immune neurological diseases like ischemic or hemorrhagic stroke. This study aimed to measure the urine and serum IL-37 levels in patients with acute ischemic stroke. Method: Twelve patients consented for the study. Two sets of serum and urine samples were obtained and analyzed; one upon admission to the hospital, and the second the next morning after overnight fasting. The trends in serum level of IL-37 in 5 stroke patients, while trends in urine level of 6 patients were available, measured by real-time polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). Prior studies with healthy volunteers as control group have consistently showed IL-37 plasma level around or less than 65 pg/ml with maximum normal levels on ELISA approximated at 130 pg/ml. Results: IL-37 level in urine in stroke patients ranged from 297 - 4467. IL-37 levels were in the range of 300s to 1000s in patients with ischemic stroke compared with reported healthy controls in literature where the level was always less than 90. Three of these 10 patients presented within 3 hours of stroke onset with IL-37 serum levels being 2655 pg/ml, 3517 pg/ml and 5235 pg/ml. In all others, it ranged much less than that, with the trend of delayed presentation giving less IL-37 levels, both in urine and serum. There were no clear differences found in patients with or without tPA, diabetes, hyperlipidemia and high blood pressure in our small study. Conclusion: The study shows a rather stable elevation of IL-37 levels post-ischemic stroke, which if compared to available data from other studies, is 3-10 times elevated after acute ischemic stroke with an uptrend in the first few days. IL-37 plays some role in mediating post-stroke inflammation with significant rise in serum and urine IL-37 levels suggesting a key role of this novel cytokine in post-stroke pathology. This is the first ever reported study measuring and trending IL-37 levels in human plasma after an acute ischemic stroke.


Stroke ◽  
2017 ◽  
Vol 48 (suppl_1) ◽  
Author(s):  
Jiang He ◽  
Yonghong Zhang ◽  
Tan Xu ◽  
Dali Wang ◽  
Chung-Shiuan Chen ◽  
...  

Although elevated blood pressure (BP) is very common in patients with acute ischemic stroke, the management of hypertension among them remains controversial. We tested the effect of immediate BP reduction on two-year mortality and major disability in acute ischemic stroke patients. The China Antihypertensive Trial in Acute Ischemic Stroke, a randomized, single-blind, blinded end-points trial, was conducted in 4,071 patients with ischemic stroke within 48 hours of onset and elevated systolic BP (SBP). Patients were randomly assigned to receive antihypertensive treatment (n=2,038) or to discontinue all antihypertensive medications (n=2,033) during hospitalization. Post-treatment follow-ups were conducted at 3, 12, and 24 months after hospital discharge. The primary outcome was a composite of death and major disability at the two-year follow-up visit. Mean SBP was reduced by 21.8 in the treatment group and 12.7 mm Hg in the control group within 24 hours after randomization (P<0.001). Mean SBP was 137.3 mm Hg in the treatment group and 146.5 in the control group at day 7 after randomization (P<0.001). At two-year follow-up, study outcomes were obtained in 1945 (95.4%) participants in the treatment group and 1925 (94.7%) in the control group. 78.8% of the patients in the treatment group and 72.6% in the control group reported the use of antihypertensive medications (p<0.001). SBP was 138.8 mmHg in the antihypertensive treatment group and 139.7 in the control group (p=0.02). Among patients in the antihypertensive treatment group, 24.5% (476/1945) died or had a major disability, compared with 22.1% (425/1925) in the control group (odds ratio 1.14 [95% CI 0.99 to 1.33], p=0.078). Hazard ratios for all-cause mortality (1.01 [0.81, 1.25], p=0.95), recurrent stroke (0.91 [0.73, 1.13], p=0.40), and vascular events (0.97 [0.79, 1.19], p=0.76) were not statistically significant comparing the antihypertensive treatment group to the control group. The effect of antihypertensive treatment did not differ by pre-defined subgroups. In conclusion, among patients with acute ischemic stroke, BP reduction with antihypertensive medications during hospitalization did not reduce or increase the composite outcome of death and major disability over two years.


2020 ◽  
Vol 2020 ◽  
pp. 1-6
Author(s):  
Zhi-Xin Huang ◽  
Jianguo Lin ◽  
Cheng Zhang ◽  
Ying-Yi Dai ◽  
Songbin Lin ◽  
...  

Early neurologic deterioration (END) in the acute phase of ischemic stroke is a serious clinical event, which is closely related to poor prognosis. Therefore, it is important to identify presentation features that predict END and take relevant treatment measures, as they could help to prevent the deterioration of high-risk patients. The prospective intervention study was carried out from January 2018 to December 2019. We included consecutive patients hospitalized for acute ischemic stroke (AIS) within 6 hours of onset. Patients were randomly assigned (1 : 1) to recanalization therapy plus Huoxiang Zhengqi Pill (HXZQ) (intervention group) or standard recanalization therapy alone (control group). The primary outcome was the development of END according to predefined criteria within the first 1 week of stroke onset. Poisson regression was used to identify predictors for END. Of the 155 patients enrolled in the study (age, 63 ± 11 years; 28.4% female), 20 (12.9%) developed END. Univariate analysis showed that the use of HXZQ and Essen stroke risk score (ESRS) (low risk group) were protective factors for END, while advanced age was a risk factor for END. However, in multivariate analysis, only ESRS (OR, 0.232; 95%CI, 0.058–0.928; P=0.039) and the use of HXZQ (OR, 0.297; 95%CI, 0.096–0.917; P=0.035) were statistically significant. ESRS can be used as the prediction factor of END. HXZQ has small side effects and wide indication. It could be used in the treatment of AIS.


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