Current Neurovascular Research - Volume 19, Issue 3, 2022

Background: Ischemic brain injury often results in irreversible pyroptosis of neurons. Sevoflurane (Sevo) post-treatment exerts an alleviative role in neuroinflammation. Objectives: This work evaluated the mechanism of Sevo post-treatment in oxygen-glucose deprivation (OGD)-induced pyroptosis of rat hippocampal neurons. Methods: Rat hippocampal neuron cell line H19-7 cells were treated with OGD, followed by posttreatment of 2% Sevo. The expression patterns of Mafb ZIP Transcription Factor B (Mafb) and dual- specificity phosphatase 14 (DUSP14) were determined via quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting methods. H19-7 cell viability and the release of lactate dehydrogenase (LDH) were examined via the cell counting kit-8 and LDH assay kits. Levels of pyroptosis-related proteins and cytokines NOD-like receptor family, pyrin domain containing 3 (NLRP3), N-term cleaved Gasdermin-D (GSDMD-N), cleaved-caspase-1, interleukin (IL)-1β, and IL-18 were also examined. The binding relation between Mafb and the DUSP14 promoter was detected. Besides, the roles of Mafb/DUSP14 in OGD-induced pyroptosis of rat hippocampal neurons were investigated through functional rescue experiments. Results: Mafb and DUSP14 expression levels were decreased in OGD-induced hippocampal neurons. Sevo post-treatment up-regulated Mafb and DUSP14, facilitated H19-7 cell viability, inhibited LDH release, and reduced levels of NLRP3, GSDMD-N, cleaved-caspase-1, IL-1β, and IL-18. Mafb increased DUSP14 expression via binding to the DUSP14 promoter. Repressing Mafb or DUSP14 exacerbated pyroptosis of hippocampal neurons. Conclusion: Sevo post-treatment increased Mafb and DUSP14 expressions, which repressed OGDinduced pyroptosis of hippocampal neurons.

Background: Single Small Subcortical Infarction (SSSI) is an isolated small infarction in the territory of perforating artery with a maximum diameter of 20 mm in axial Diffusion-Weighted Imaging (DWI). About 20 to 30% of SSSI patients were reported to have Early Neurological Deterioration (END) in the acute phase, which brought adverse effects on long-term outcomes. The effect of the alteplase on the outcome of SSSI, especially END and long-term outcomes, was ambiguous. Objective: The study aims to find out the efficacy and safety of intravenous recombinant tissue Plasminogen Activator (rt-PA) on long-term and short- outcomes of patients with SSSI as compared to patients who received standard medical care. Methods: The patients were retrospectively screened from a stroke registry of the neurology department of 1st Affiliated Hospital of Zhengzhou University from January 2013 to December 2020. Based on treatment modality, patients were dichotomized into alteplase and standard medical care groups. To minimize confounding factors in subgroups, a propensity score matching analysis was done. The primary outcome was the favorable functional outcome 3 months after stroke onset, defined by attaining a score of ≤2 points on the modified Rankin scale (mRS), secondary outcome was the prevention of occurrence of END, defined as an increase of ≥2 points in the total score or ≥1point on motor subunit in the National Institutes of Health Stroke Scale (NIHSS) score within 72 hours after admission, safety features were symptomatic intracranial hemorrhage (sICH) or death. Multivariate analysis was employed to find the efficacy and safety of alteplase in the treatment of SSSI. Results: A total of 717 patients with anterior circulation SSSI were selected, and 132 were included in the final analysis. Forty-five patients were treated with alteplase within 4.5 hours and 87 with standard medical care, and 44 pairs were successfully matched by propensity score. Pre-match data showed that the alteplase thrombolysis group showed a higher proportion of favorable outcomes at 3-month follow-up [OR=0.315, 95%CI:0.106, 0.931, P = 0.037] but did not reduce the incidence of END compared with the non-thrombolytic group [OR = 1.033, 95%CI:0.417,2.554, P = 0.943]. Post-match data showed that the alteplase group also showed a higher proportion of favorable outcomes at 3-month follow-up [OR = 0.247, 95%CI: 0.074, 0.830, P = 0.024]; however, it did not reduce the incidence of END compared with the non-thrombolytic group [OR = 1.241, 95%CI: 0.433,3.554, P = 0.688]. There was one case of asymptomatic ICH in alteplase treated patients. Conclusion: Patients with SSSI in the anterior circulation are more likely to achieve 3 months favorable outcomes than those who were treated with standard medical care; however, treatment with alteplase may not prevent the occurrence of END.

Introduction: Depression is a class of important mental illness, which has become a severe health problem perplexing the world due to its high morbidity rate, high disability rate, and great disease burden. This study aimed to evaluate the role and possible mechanisms of P2RY12 in the depression-like behaviors model. Methods: Serum samples of patients with depression-like behaviors were used to analyze the expression of P2RY12. Models of mice were given LPS via intraperitoneal injection for 7 days. Behavioral tests were executed in this experiment. Results: The expression of P2RY12 in models of depression-like behaviors or mice with depression- like behaviors were induced. The inhibition of P2RY12 presents depression-like behaviors and reduces inflammation in the model of depression-like behaviors. P2RY12 induced NLRP3 expression and suppressed NLRP3 ubiquitination in a model of depression-like behavior. The inhibition of NLRP3 reduced the effects of P2RY12 in mice model of depression-like behaviors. The regulation of NLRP3 controlled the effects of the P2RY12 in vitro model of depression-like behaviors. Conclusion: We conclude that P2RY12 increased neuroinflammation to accelerate depression-like behaviors by NLPR3 inflammasome, providing novel information for the treatment of depressionlike behaviors.

Objective: This study aimed to explore and analyze the effect of acupuncture on improving the enteral nutrition level and gastrointestinal dynamics in patients who had suffered a severe stroke. Methods: A total of 122 patients who experienced a severe stroke who were treated in the intensive care unit of the Affiliated Hospital of Hebei University (China) between September 2021 and March 2022 were randomly divided into two groups as follows: 1) the observation group, the participants of which received acupuncture combined with early enteral nutrition (61 cases); 2) the control group, the participants of which received early enteral nutrition (61 cases). Following treatment, the hemoglobin, neutrophil count, blood glucose, albumin, pre-albumin, immediate postprandial antral area, antral contraction frequency (at 2 min), and antral motility index on days 1 and 7 of treatment were compared between the two groups. Results: The total clinical effective rate was 96.72% in the observation group and 77.05% in the control group. The curative effect comparison between the two groups after seven days of treatment showed a lower probability of gastrointestinal bleeding, faster recovery of gastrointestinal motility, and a higher level of nutrient absorption in the observation group. Serum albumin, pre-albumin, hemoglobin, total lymphocyte count, immediate postprandial maximum (max) and minimum (mix) area of the gastric antrum, antral contraction frequency (at 2 min), and antral motility index were higher in the observation group than in the control group (P < 0.05). The difference in blood glucose levels between the two groups was not statistically significant (P > 0.05). Conclusion: Acupuncture improved the enteral nutrition status of patients who had suffered a severe stroke and promoted gastrointestinal motility. The combination of acupuncture and early enteral nutrition could reduce damage to the gastrointestinal mucosal barrier caused by stress, changes in metabolism, and improved gastrointestinal function.

Background and Objective: The potential impact of rebleeding and Delayed Cerebral Ischemia (DCI) on long-term survival in patients with aneurysmal subarachnoid hemorrhage (aSAH) remained unclear. This study aimed to investigate whether DCI and rebleeding increase the risk of long-term all-cause mortality in patients with aSAH who survived the follow-up period of one year. Methods: We retrospectively collected data on patients with atraumatic aSAH who were still alive 12 months after aSAH occurrence between December 2013 and June 2019 from the electronic health system. Patients were then classified by the occurrence of rebleeding or DCI during hospitalization. Death records were obtained from an administrative database, the Chinese Household Registration Administration System, until April 20, 2021. Multivariable Cox proportional hazards models were used to compare overall survival in different groups. Sensitivity analysis was performed with propensity-score matching (PSM). Results: A total of 2,607 patients were alive one year after aSAH. The crude annual death rate from any cause among patients who had rebleeding (7.2 per 100 person-years) and patients who had DCI (3.7 per 100 person-years) during hospitalization was higher than that of patients with neither event (2.1 per 100 person-years). Multivariate analysis showed that rebleeding is an independent risk factor for long-term mortality (adjusted hazard ratio (aHR), 2.37; 95% confidence interval (CI), 1.47- 3.81). DCI was an independent prognostic factor of poorer overall survival (aHR, 2.09; 95% CI, 1.54-2.84). Conclusion: Amongst patients alive one year after aSAH, rebleeding and DCI during hospitalization were independently associated with higher rates of long-term mortality.

Background: Chemotherapy-induced peripheral neuropathy is a debilitating pain syndrome produced as a side effect of antineoplastic drugs like paclitaxel. Despite efforts, the currently available therapeutics suffer from serious drawbacks like unwanted side effects and poor efficacy and provide only symptomatic relief. Hence, there is a need to find new therapeutic alternatives for the treatment of chemotherapy-induced peripheral neuropathy. Objective: The objective of this study was to explore the protective potential of caffeic acid phenethyl ester in paclitaxel-induced neuropathic pain. Methods: We examined the effects of caffeic acid phenethyl ester by administering paclitaxel (2 mg/kg, intraperitoneal) to female Sprague Dawley rats on four alternate days to induce neuropathic pain, followed by the administration of caffeic acid phenethyl ester (10 and 30 mg/kg, intraperitoneally). Results: Rats that were administered paclitaxel showed a substantially diminished pain threshold and nerve functions after 28 days. A significantly increased protein expression of Wnt signalling protein (β-catenin), inflammatory marker (matrix metalloproteinase 2) and a decrease in endogenous antioxidant (nuclear factor erythroid 2–related factor 2) levels were found in paclitaxel administered rats in comparison to the naïve control group. Caffeic acid phenethyl ester (10 and 30 mg/kg, intraperitoneal) showed improvements in behavioural and nerve function parameters along with reduced expression of β-catenin, matrix metalloproteinase 2 and an increase in nuclear factor erythroid 2– related factor 2 protein expression. Conclusion: The present study suggests that caffeic acid phenethyl ester attenuates chemotherapyinduced peripheral neuropathy via inhibition of β-catenin and matrix metalloproteinase 2 and increases nuclear factor erythroid 2–related factor 2 activation.

Background and Purpose: The purpose of this study was to show dynamic changes in carotid and vertebral artery using carotid Doppler ultrasonography (DUS) through a long-term follow- up exam, and determine their associations with stroke recurrence. Methods: We consecutively enrolled stroke or transient ischemic attack (TIA) patients who had undergone DUS more than twice with intervals of three months or more. Stroke recurrence during follow-up was also investigated by reviewing medical records. Progress or regress of plaque was defined as more than 0.1 mm change from the initial scan with a semi-quantitative measurement. The development of new plaque was also regarded as plaque progress. Increased intima-media thickness and plaque presence were interpreted at the initial and follow-up scans. Factors related to progression or regression were analyzed. The relationship between plaque change and stroke recurrence was investigated. Results: A total of 201 patients were enrolled (186 ischemic stroke patients and 15 TIA patients). There were 61 (30.3%) females. Their mean age was 64.2 ± 9.9 years. During a follow-up of 35.0 ± 22.6 (mean ± SD) months, plaque progress was observed in 92 (45.8%) and plaque regress in 13 (6.5%). Stroke recurred in 18 patients. Plaque progression showed no significant association with age, risk factors, statin use, or subtype. After adjustment of age, sex, diabetes, and stroke subtype, multiple logistic regression showed a significant association of plaque progression with stroke recurrence (odds ratio: 3.8, 95% confidence interval: 1.1 to 13.1, p = 0.034). Patients with plaque regress were significantly younger than those without plaque regress (57.8 years vs. 64.6 years, p = 0.041). Conclusion: Plaque progression occurred in 46% of stroke or TIA patients. Plaque progression was significantly associated with clinical stroke recurrence. Plaque regressed in 6.5% of patients. Patients with regression were younger than those without.

Objective: We investigated the factors associated with cerebrospinal fluid (CSF) flow artifacts on fluid-attenuated inversion recovery imaging in patients with carotid artery (CA) stenosis. Methods: Each CSF artifact grade was defined by comparing the highest intensity in a given region of interest (ROI) to those in reference ROIs, as follows: higher than the intensity of normal white matter in the centrum semiovale = 2 points; equal to or less than the white matter, and higher than CSF = 1 point; and equal to CSF = 0. CSF flow scores in eight sites were measured and added to the total score (0 -16). The prevalences of each finding, specifically white matter lesions, CA stenoses and brain atrophy, were compared using multivariate logistic regression models. Results: We evaluated the findings in 54 patients with CA stenosis treated by CA stenting (CAS) and 200 adults with no history of neurological disorders (control group). Adjusted by stroke risk factors, a CSF flow score ≤ 11 was positively associated with CA stenosis, heart rate > 70 / min, and brain atrophy, and negatively with the female gender. The score was 12.8 ± 1.8 in the control group and 12.0 ± 2.0 in CA stenosis group after CAS, which was significantly higher than before CAS (10.4 ± 2.8, p<0.001). Conclusion: The CSF flow score was associated with female gender, brain atrophy, heart rate, and severe CA stenosis, and was found to be elevated after revascularization.

Background: Previous studies have demonstrated that statins can relieve inflammatory brain injury after intracerebral hemorrhage (ICH), but the mechanisms remain poorly characterized. This study aims to test whether simvastatin exerts an anti-inflammatory effect by regulating the proresolving mediators. Methods: First, male Sprague–Dawley rats had an injection of 200 μL autologous blood. Then, rats were randomly divided into groups treated with simvastatin (i.p. 2 mg/kg) or vehicle. Next, all rats underwent pro-resolving mediator lipoxin A4 (LXA4) level detection, flow cytometric, immunofluorescence, brain edema measurement, neurological scoring and western blot analysis. Results: We found that simvastatin significantly increased the plasma level of LXA4, an endogenous formyl-peptide receptor 2 (FPR2) agonist, in the early stage of ICH. Consistent with the effect of simvastatin, exogenous LXA4 administration also promoted apoptosis of the circulating neutrophils, reduced neutrophils brain infiltration, and ameliorated inflammatory brain injury after ICH. In addition, similar to simvastatin, exogenous LXA4 markedly decreased the level of phosphorylated p38 mitogen-activated protein kinase (MAPK) and the apoptosis-related proteins myeloid cell leukemia 1(Mcl-1)/Bax ratio (a decreased ratio represents the induction of apoptosis) in circulating neutrophils isolated from ICH rats. Notably, all of the aforementioned effects of simvastatin on ICH were significantly abolished by Boc-2, a selective antagonist of FPR2. Moreover, simvastatin led to a similar Mcl-1/Bax ratio reduction as SB203580 (a p38 MAPK inhibitor), but it was abolished by P79350 (a p38 MAPK agonist). Conclusion: Collectively, these results suggest that simvastatin ameliorates ICH-mediated inflammatory brain injury, possibly by upregulating the level of pro-resolving mediator LXA4 and further stimulating the FPR2/p38 MAPK signaling pathway.

Background: Detection or monitoring of brain damage is a clinically crucial issue. Nucleic acids in the whole blood can be used as biomarkers for brain injury. Polymerase chain reaction (PCR) which is one of the most commonly used molecular diagnostic assays requires isolated nucleic acids to initiate amplification. Currently used nucleic acid isolation procedures are complicated and require laboratory equipments. Objective: In this study, we tried to develop a simple and convenient method to isolate nucleic acids from the whole blood sample using a tiny battery-powered electric device. The quality of the isolated nucleic acids should be suitable for PCR assay without extra preparation. Methods: A plastic device with separation chamber was designed and printed with a 3D printer. Two platinum electrodes were placed on both sides and a battery was used to supply the electricity. To choose the optimal nucleic acid isolation condition, diverse lysis buffers and separation buffers were evaluated, and the duration and voltage of the electricity were tested. Western blot analysis and PCR assay were used to determine the quality of the separated nucleic acids. Results: 2ul of whole blood was applied to the cathode side of the separation chamber containing 78 ul of normal saline. When the electricity at 5 V was applied for 5 min, nucleic acids were separated from segment 1 to 3 of the separation chamber. The concentration of nucleic acids peaked around 7~8 mm from cathode side. PCR assay using the separation buffer as the template was performed successfully both in conventional and realtime PCR methods. The hemoglobin in the whole blood did not show the inhibitory effect in our separation system and it may be due to structural modification of hemoglobin during electric separation. Conclusion: Our simple electric device can separate nucleic acids from the whole blood sample by applying electricity at 5 V for 5 min. The separation buffer solution taken from the device can be used for PCR assay successfully.

Background: Alzheimer’s Disease (AD) impairs memory and cognitive functions in the geriatric population and is characterized by intracellular deposition of neurofibrillary tangles, extracellular deposition of amyloid plaques, and neuronal degeneration. Literature suggests that latent viral infections in the brain act as prions and promote neurodegeneration. Memantine possesses both anti-viral and N-methyl-D-aspartate (NMDA) receptor antagonistic activity. Objectives: This research was designed to evaluate the efficacy of antiviral agents, especially valacyclovir, a prodrug of acyclovir in ameliorating the pathology of AD based on the presumption that anti-viral agents targeting the Herpes Simplex Virus (HSV) can have a protective effect on neurodegenerative diseases like Alzheimer’s disease. Methods: Thus, we evaluated acyclovir’s potential activity by in-silico computational docking studies against acetylcholinesterase (AChE), butyrylcholinesterase (BuChE), and beta-secretase 1 (BACE-1). These findings were further evaluated by in-vivo scopolamine-induced cognitive impairment in rats. Two doses of valacyclovir, a prodrug of acyclovir (100 mg/kg and 150 mg/kg orally) were tested. Results: Genetic Optimisation for Ligand Docking scores and fitness scores of acyclovir were comparable to donepezil. Valacyclovir improved neurobehavioral markers. It inhibited AChE and BuChE (p<0.001) enzymes. It also possessed disease-modifying efficacy as it decreased the levels of BACE-1 (p<0.001), amyloid beta 1-42 (p<0.001), amyloid beta 1-40 (p<0.001), phosphorylatedtau (p<0.001), neprilysin (p<0.01), and insulin-degrading enzyme. It ameliorated neuroinflammation through decreased levels of tumour necrosis factor α (p<0.001), nuclear factor-kappa B (p<0.001), interleukin 6 (p<0.001), interleukin 1 beta (p<0.001), and interferon-gamma (p<0.001). It also maintained synaptic plasticity and consolidated memory. Histopathology showed that valacyclovir could restore cellular density and also preserve the dentate gyrus. Conclusion: Valacyclovir showed comparable activity to donepezil and thus can be further researched for the treatment of Alzheimer’s disease.

Background and Aims: The aim of this study is to investigate the relationships between the original and modified total cerebral Small Vessel Disease (CSVD) score and gait and balance impairment using quantitative and semi-quantitative tests. Methods: In our study, patients aged 45 to 85 consecutively recruited. CSVD manifestations were identified with brain Magnetic Resonance Imaging (MRI), and the original and modified CSVD scores were calculated based on the results. Gait and balance function were assessed using both gait parameters and clinical rating scales. The correlation between the original and modified total scores of the CSVD and gait and balance dysfunction was demonstrated. Results: 224 patients were enrolled in the study. Gait and balance disorders were associated with both the original and modified CSVD scores. A significant association remained after adjusting for gender, height, age, hypertension, and other relevant risk factors. The binary logistic regression and chi-squared trend tests revealed that impairment of movement function significantly correlated with the modified CSVD score and that the dysfunction was significantly higher for patients with modified CSVD scores of 5-6 than those with scores of 1-2. In Receiver Operating Characteristic (ROC) analysis, modified CSVD scores were more accurate in predicting gait impairment than original CSVD scores. Conclusion: We found both original and modified total CSVD scores to be related to gait and balance disorder, and the modified CSVD score was more accurate in identifying movement impairment and should be used as an effective tool in investigating CSVD and motor dysfunction.

Aims: Although early tracheostomy (ET) is recommended for patients with severe stroke, the optimal timing of tracheostomy for patients with intracerebral haemorrhage (ICH) remains controversial. This study aimed to explore the clinical characteristics, risk factors and timing of tracheostomy in patients after tracheal intubation using a propensity-matched analysis. Methods: We conducted a retrospective database search and assessed 267 consecutive patients who underwent endotracheal intubation (175 of whom underwent tracheostomy) and ICH between July 2017 and June 2021. A logistic regression model was applied to identify the critical factors influencing the decision for tracheostomy by comparing factors in a tracheostomy group and a nontracheostomy group. Patients were divided into an early (≤5 days) or a late (>5 days) group according to the median time of tracheostomy. Propensity score matching was performed to adjust for possible confounders and investigate differences in outcomes between ET and late tracheostomy (LT). Results: Among the 267 enrolled patients with ICH and endotracheal intubation, 65.5% received tracheostomy during hospitalisation, and 52.6% received ET. The independent risk factors for tracheostomy included National Institute of Health Stroke Scale (NIHSS) (odds ratio [OR]: 1.179; 95% confidence interval [CI]: 1.028-1.351; P = 0.018), aspiration (OR: 2.171; 95% CI: 1.054-4.471; P = 0.035) and infiltrates (OR: 2.149; 95% CI: 1.088-4.242; P = 0.028). Using propensity matching, we found that ET was associated with fewer antibiotic-using days (15 vs. 18; P < 0.001) and sedativeusing days (6 vs. 8; P < 0.001), shorter intensive care unit (ICU) Length of Study (LOS) (9 vs. 12; P < 0.05) and reduced in-ICU costs (3.59 vs. 7.4; P < 0.001) and total hospital costs (8.26 vs. 11.28, respectively; P < 0.001). Muscle relaxants (31.8% vs. 60.6%) were used less frequently in patients with ET (P = 0.001). However, there were no differences between the ET and LT groups in terms of modified Rankin Scale (mRS) (4 vs. 4; P = 0.932), in-general-ward costs (4.74 vs. 4.37; P = 0.052), mechanical ventilation days (6 vs. 6; P = 0.961) and hospital LOS (23 vs. 23; P = 0.735) as well as the incidences of ventilator-associated pneumonia (28.8% vs. 37.9%; P = 0.268), tracheostomyrelated complications (16.7% vs. 19.7%; P = 0.652), respiratory failure (24.2% vs. 31.8%; P = 0.333), all-cause deaths (15.2% vs. 16.7%; P = 0.812) and pneumonia (77.3% vs. 87.9%; P = 0.108). Conclusion: We recommend ET for high-risk patients with ICH. Although ET cannot reduce inhospital mortality or improve patient prognosis, it may help reduce hospital costs and ICU LOS as well as the use of antibiotics, sedatives and muscle relaxants.