Current Neurovascular Research - Volume 20, Issue 2, 2023

Background: Cerebral small vessel disease (CSVD) refers to a common cerebrovascular disease and white matter hyperintensities (WMHs) constitute a typical feature of CSVD. However, there has not been a large number of studies investigating the relationship between lipid profile components and WMHs. Methods: Altogether, 1019 patients with CSVD were enrolled at the First Affiliated Hospital of Zhengzhou University between April 2016 to December 2021. Baseline data were collected for all patients, including demographic characteristics and clinical data. WMH volumes were evaluated by two experienced neurologists using the MRIcro software. Multivariate regression analysis was used to investigate the relationship among the severity of WMHs, blood lipids and common risk factors. Results: Altogether, 1019 patients with CSVD were enrolled, including 255 in the severe WMH group and 764 in the mild WMH group. After including age, sex and blood lipids to construct a multivariate logistic regression equation, we observed that the severity of WMHs was independently predicted by low-density lipoprotein (LDL), homocysteine level and history of cerebral infarction. Conclusion: We used WMH volume, a highly accurate measure, to assess its relationship with lipid profiles. The WMH volume increased with a decrease in LDL. This relationship was more significant, especially among the subgroups of patients aged <70 years and men. Patients with cerebral infarction and higher homocysteine levels were more likely to have higher WMH volumes. Our study has provided a reference for clinical diagnosis and therapy, especially for discussing the role of blood lipid profiles in the pathophysiology of CSVD.

Background: Cortical laminar necrosis (CLN) is a specific type of cortical infarction, and little is known about its frequency and outcomes. We aimed to investigate the prevalence and outcomes of CLN caused by brain infarction and its prognostic factors. Methods: This retrospective cohort study included patients with acute ischemic stroke (AIS) between 2019 and 2022 and for whom magnetic resonance images obtained at our center showed acute-stage CLN. Their medical records were collected and analyzed. An unfavorable outcome was defined as a modified Rankin Scale score of 3-6 at 90 days. Logistic regression was performed to identify independent predictors of an unfavorable outcome. Results: Among 5548 consecutive patients with AIS, 151 patients (2.7%) were diagnosed with CLN, and 112 had CLN enrolled in the final analysis. At 90 days, 25 patients (22.3%) had an unfavorable outcome. Compared with the favorable group, poor outcome patients had higher rates of previous stroke (p = 0.012), higher National Institutes of Health Stroke Scale (NIHSS) scores at admission (p < 0.001), and were more likely to have early neurologic deterioration (END) (p = 0.014), diffuse ischemic lesions (p = 0.011), and lesions involving multiple lobes (p = 0.030). In multivariable analysis, the initial NIHSS score (OR, 1.258, (95% CI 1.090 - 1.453), p = 0.002) and END (OR, 5.695, [95% CI 1.410 - 23.007], p = 0.015) were independently associated with unfavorable outcome. Conclusion: CLN is a rare ischemic event but has a good prognosis in most cases. A higher initial NIHSS score and END may predict an unfavorable outcome.

Objective: White matter hyperintensity (WMH) is related to the increased risk of ischemic stroke. It is unclear if H-type hypertension (H-type HBP) is associated with periventricular WMH (PWMH) and deep WMH (DWMH) of acute ischemic stroke. This study investigated the correlation between H-type HBP and the severity of PWMH and DWMH in acute ischemic stroke. Methods: Consecutive patients with acute ischemic stroke were included in this cross-sectional observational study. The patients were divided into the following groups: the normal group, the simple hypertension group (Simple HBP), the simple hyperhomocysteinemia group (Simple HHcy) and the H-type HBP group. MR imaging and relevant clinical variables were obtained from the medical records. PWMH and DWMH were rated by using the Fazekas scale (score 0-3). All patients were defined to have moderate-severe PWMH or DWMH (score 2-3) and no or mild group (score 0-1). Multivariate binary logistic regression analysis was performed to determine the relationship between H-type HBP and the severity of PWMH and DWMH. Results: Among 542 patients, 227 had moderate-severe PWMH and 228 had moderate-severe DWMH. Compared to the no or mild group, patients with moderate-severe PWMH (median age: 73 vs. 63 years) and DWMH (median age: 70 vs. 65.5 years) were older. Compared to the no or mild group, moderate-severe PWMH and DWMH were associated with a history of ischemic stroke (moderate-severe PWMH vs. no or mild group 20.7% vs. 11.7%, p = 0.004#155;moderatesevere DWMH vs. no or mild group 20.2% vs. 12.1%, p = 0.010); We found that H-type HBP was an independent risk factor for PWMH (OR 2.64, 95% CI 1.34-5.21) and DWMH (OR 3.64, 95% CI 1.82-7.26) after adjusting for the effect of relevant risk factors. Conclusion: This study suggests that H-type HBP is associated with the severity of PWMH and DWMH in acute ischemic stroke patients, which deserves further prevention measures.

Background: NOD-like receptor protein 3 (NLRP3) inflammasome-mediated pyroptosis is strongly related to cerebral ischemia/reperfusion (I/R) injury. DDX3X, the DEAD-box family's ATPase/RNA helicase, promotes NLRP3 inflammasome activation. However, whether DDX3X deficiency attenuates NLRP3 inflammasome-mediated pyroptosis induced by cerebral I/R injury. Objectives: This study investigated whether DDX3X deficiency attenuates NLRP3 inflammasomemediated pyroptosis in N2a cells after oxygen-glucose deprivation/ reoxygenation (OGD/R) treatment. Methods: In vitro model of cerebral I/R injury, mouse neuro2a (N2a) cells subjected to OGD/R were treated with the knockdown of DDX3X. Cell counting kit-8 (CCK-8) assay and Lactate dehydrogenase (LDH) cytotoxicity assay were conducted to measure cell viability and membrane permeability. Double immunofluorescence was performed to determine the pyroptotic cells. Transmission electron microscopy (TEM) was used to observe morphological changes of pyroptosis. Pyroptosis-associated proteins were analyzed by Western blotting. Results: The OGD/R treatment reduced cell viability, increased pyroptotic cells and released LDH compared to the control group. TEM showed membrane pore formation of pyroptosis. Immunofluorescence showed that GSDMD was translocated from the cytoplasm to the membrane after OGD/R treatment. Western blotting showed that the expression of DDX3X, and pyroptosis-related proteins (NLRP3, cleaved-Caspase1, and GSDMD-N) were increased after OGD/R treatment. Nevertheless, DDX3X knockdown markedly improved cell viability and reduced LDH release, expression of pyroptosis-related proteins, and N2a cells pyroptosis. DDX3X knockdown significantly inhibited membrane pore formation and GSDMD translocation from cytoplasm to membrane. Conclusion: This research demonstrates for the first time that DDX3X knockdown attenuates OGD/R-induced NLRP3 inflammasome activation and pyroptosis, which implies that DDX3X may become a potential therapeutic target for cerebral I/R injury.

Background: To investigate the combined effect of red blood cell distribution width (RDW) and inflammatory biomarkers on in-hospital outcomes of acute ischemic stroke(AIS) patients with thrombolysis. Methods: 417 AIS patients with thrombolysis were included. The participants were divided into four groups according to the cut-off of white blood cell (WBC) or C reactive protein (CRP) and RDW: LWLR, LWHR, HWLR, and HWHR; or LCLR, LCHR, HCLR, and HCHR (L-low, Hhigh, W-WBC, C-CRP, R-RDW). Logistic regression models were used to calculate the odds ratios (ORs) and 95% confidence intervals (CIs) of in-hospital pneumonia and functional outcome across the four subgroups. Results: Patients with higher RDW and inflammatory biomarkers levels have the highest risk of in-hospital outcomes. Compared with patients in the LWLR group, the ORs (95% CIs) of those in the HWHR group were 12.16 (4.21-35.14) and 9.31 (3.19-27.17) for in-hospital pneumonia and functional outcome. The ORs (95% CIs) of those in the HCHR group were 6.93 (2.70-17.78) and 3.38 (1.10-10.39) for in-hospital pneumonia and functional outcome, compared with patients in the LCLR group. Simultaneously adding RDW and WBC or CRP to the basic model with established risk factors significantly improved risk discrimination and reclassification for pneumonia and functional outcome (all p <0.05). Conclusions: Combined RDW and inflammatory biomarkers within 4.5 hours had a better predictive power for in-hospital outcomes of AIS patients with thrombolysis.

Objective: The objective of this study is to study the mechanism of Low frequency electrical stimulation (LFS) in the treatment of drug-resistant epilepsy by regulating the protein kinase A (PKA)-cAMP response element-binding protein (CREB) signaling pathway upstream of gamma aminobutyric acid A (GABAA) receptor. Methods: Primary hippocampal neurons were extracted and cultured from fetal rat brains and randomly divided into the normal control group, PKA-CREB agonist group, and PKA-CREB inhibitor group. Drug-resistant epileptic rats were established and randomly divided into the pharmacoresistant group, LFS group, PKA-CREB agonist combined with hippocampal LFS group, and PKA-CREB inhibitor combined with hippocampal LFS group. The normal rats were in the normal control group and drug-sensitive rats were in the pharmacosensitive group. The seizure frequency of epileptic rats was determined using video surveillance. The expression of PKA, CREB, p-CREB, and GABAA receptor subunits α1 and β2 of each group were detected using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blotting assays. Results: The in vitro expression levels of PKA, CREB, and p-CREB in the agonist group were significantly higher than those in the normal control group (NRC group), while the expression levels of GABAA receptor subunits α1 and β2 were significantly lower than those in the NRC group. The expression levels of PKA, CREB, and p-CREB in the inhibitor group were significantly lower, while the expression levels of GABAA receptor subunits α1 and β2 were significantly higher than those in the NRC group. The in vivo seizure frequency was significantly lower in the LFS group than in the pharmacoresistant group (PRE group). Compared to the LFS group, the seizure frequency and the expression levels of PKA, CREB, and p-CREB in the rat hippocampus were significantly higher, and the expression levels of GABAA receptor subunits α1 and β2 were significantly lower in the agonist group. The results in the inhibitor group were exactly the opposite of those in the agonist group. Conclusion: The PKA-CREB signaling pathway is involved in the regulation of GABAA receptor subunits α1 and β2. In addition, LFS plays an important role in increasing GABAA receptor expression by regulating the PKA-CREB signaling pathway.

Background: Insulin resistance is a phenomenon in which the lowering blood glucose capacity of insulin is decreased, which is a feature of type 2 diabetes mellitus. Some previous studies have found an association between insulin resistance and migraine. The triglyceride glucose (TyG) index is used to assess insulin resistance. However, there is no report on the association between the TyG index and migraine. Objective: We present a cross-sectional study from the National Health and Nutrition Examination Survey (NHANES) to clarify the association between the TyG index and migraine. Methods: Data was acquired from the NHANES. Migraine was diagnosed based on patient selfreport and prescription medication. Data were analyzed using the weighted linear regression model, weighted chi-square test, logistic regression models, smooth curve fittings, and the twopiecewise linear regression model. Empower software was used for all data analysis. Results: A total of 18704 participants were enrolled in this study, of which 209 were migraineurs. The rest were set as control. There was a statistically significant difference in mean age (p = 0.0222), gender (p < 0.0001), distribution of race (P < 0.0001), and drug usage between the two groups. However, there were no differences in type 2 diabetes mellitus, type 1 diabetes mellitus, total cholesterol, triglycerides, glucose, and TyG index between the two groups. According to logistic regression models, there was a linear relationship between TyG index and migraine in model 3 (odds ratio (OR = 0.54, p = 0.0165). particularly in female (OR= 0.51, p = 0.0202) or Mexican American (OR= 0.18, p = 0.0203). Moreover, there was no inflection point between the TyG index and migraine. Conclusion: In conclusion, there was a linear relationship between the TyG index and migraine. A higher TyG index predicts a lower incidence of migraine, particularly in females and Mexican Americans. Meanwhile, there is no inflection point between the TyG index and migraine.

Background: Previous studies have shown that the neurological damage caused by middle cerebral artery occlusion (MCAO) is not only limited to local infarction but can also cause secondary damage in distant sites, such as the hypothalamus. 5-hydroxytryptamine (5-HT)/ 5-HT transporter (5-HTT) and 5-HT receptor 2A (5-HT2A) are important in the treatment of cerebrovascular diseases. Objective: This study aimed to study the effect of electroacupuncture (EA) on the expression of 5- HT, 5-HTT, and 5-HT2A in the hypothalamus of rats with ischemic brain injury and to explore the protective effect and potential mechanism of EA on the secondary injury of cerebral ischemia. Methods: Sprague-Dawley (SD) rats were randomly divided into three groups: sham group, model group, and EA group. The permanent middle cerebral artery occlusion (pMCAO) method was used to induce ischemic stroke in rats. In the EA group, the Baihui (GV20) and Zusanli (ST36) points were selected for treatment, which was administered once per day for two consecutive weeks. The neuroprotective effect of EA was evaluated by nerve defect function scores and Nissl staining. The content of 5-HT in hypothalamus was detected by enzyme linked immunosorbent assay (ELISA), and the expression of 5-HTT and 5-HT2A were detected by Western blot. Results: Compared with that in the sham group, the nerve defect function score in the model group rats was significantly increased, the hypothalamus tissue showed obvious nerve damage, the levels of 5-HT and the expression of 5-HTT were significantly reduced, and the expression of 5-HT2A was significantly increased. After 2 weeks of EA treatment, the nerve defect function scores of pMCAO rats were significantly reduced, the hypothalamic nerve injury was significantly reduced, the levels of 5-HT and the expression of 5-HTT were significantly increased, and the expression of 5-HT2A was significantly decreased. Conclusion: EA has a certain therapeutic effect on hypothalamic injury secondary to permanent cerebral ischemia, and its potential mechanism may be closely related to the upregulation of 5-HT and 5-HTT expression and the downregulation of 5-HT2A expression.

Objectives: To assess the association between carotid artery plaques and the risk of incident intracerebral hemorrhage (ICH) event in high-risk individuals for stroke. Methods: We conducted a population-based cohort study using the longitudinal participant-level data of a multicenter, cross-sectional survey in southwestern China. 2644 high-risk participants for stroke were enrolled in the year 2015. The primary outcome was new-onset ICH events during a five-year follow-up period. Multivariate logistic regression was performed to identify the association between carotid plaque and new-onset ICH. Stratified analyses and interaction tests were conducted to identify variables that might modify the association between vulnerable carotid plaque and ICH. Results: Among 2644 high-risk individuals enrolled, carotid plaques were found in 904 (34.2%) subjects, including 479 (18.1%) with stable plaques and 425 (16.1%) with vulnerable plaques. During a five-year follow-up period, 22 (0.83%) participants developed ICH. Vulnerable carotid plaque was associated with an increased risk of new-onset ICH in multivariable analyses (adjusted RR 3.72, 95 % CI 1.32 to 10.46, p=0.013). Stratified analyses and interaction analyses demonstrated the association between vulnerable carotid plaque and ICH was not changed by age, family history of stroke, hemorrhagic stroke and chronic disease, smoking, drinking, physical activity, BMI, antihypertensives, and antithrombotic drugs (all p for interaction>0.05). However, among the female cohort, participants with vulnerable plaques had a significantly higher risk of ICH compared with participants without vulnerable plaques (crude RR=9.8; 95%CI: 3.1-31.3, p<0.001; adjusted RR=26.3, 95%CI: 5.5-124.5, p<0.001), but not in man (p>0.05). Conclusion: In Chinese individuals at high risk of stroke, vulnerable carotid artery plaques are associated with an increased risk of intracerebral hemorrhage independent of classical vascular risk factors, especially in female individuals.

Introduction: Endovascular treatment (EVT) performed in the early time window has been shown to decrease the incidence of malignant middle cerebral artery infarction (MMI). However, the incidence of MMI in patients undergoing EVT during the late time window is unclear. This study aimed to investigate the prevalence of MMI in patients undergoing late EVT and compare it with that in patients undergoing early EVT. Methods: We retrospectively analyzed consecutive patients with anterior large vessel occlusion stroke who underwent EVT at Xuanwu Hospital between January 2013 and June 2021. Eligible patients were divided into early EVT (within 6 h) and late EVT (6-24 h) groups according to the time from their stroke onset to puncture and compared. The occurrence of MMI post-EVT was the primary outcome. Results: A total of 605 patients were recruited, of whom 300 (50.4%) underwent EVT within 6 h and 305 (49.6%) underwent EVT within 6-24 h. A total of 119 patients (19.7%) developed MMI. 68 patients (22.7%) in the early EVT group and 51 patients (16.7 %) in the late EVT group developed MMI (p = 0.066). After adjusting for covariate variables, late EVT was independently associated with a lower incidence of MMI (odds ratio, 0.404; 95% confidence interval, 0.242-0.675; p = 0.001). Conclusion: MMI is not an uncommon phenomenon in the modern thrombectomy era. Compared with the early time window, patients selected by stricter radiological criteria to undergo EVT in the late time window are independently associated with a lower incidence of MMI.

Introduction: The aim is to establish a rat model of infraorbital neuroinflammation with less trauma, stable pain, and a long duration of pain. The pathogenesis of TN is not fully clear. There are various models of TN in rats with different disadvantages, such as damaging the surrounding structures and inaccuracy of location for infraorbital nerve (ION). We aim to establish a rat model of infraorbital neuroinflammation with minimal trauma, a simple operation, and accurate positioning under CT guidance to help us study the pathogenesis of trigeminal neuralgia. Methods: Thirty-six adult male Sprague Dawley rats (180-220 g) were randomly divided into 2 groups and injected with talc suspension or saline through the infraorbital foramen (IOF) under CT guidance. Mechanical thresholds were measured in the right ION innervation region of 24 rats over 12 postoperative weeks. At 4 weeks, 8 weeks, and 12 weeks after the operation, the inflammatory involvement of the surgical area was evaluated by MRI, and neuropathy was observed using a transmission electron microscope (TEM). Results: The talc group had a significant decrease in the mechanical threshold at 3 days after surgery that continued until 12 weeks post-operation, and the talc group had a significantly lower mechanical threshold than the saline group 10 weeks post-operation. The talc group had significantly impaired trigeminal nerve (TGN) myelin after 8 weeks post-operation. Conclusion: The rat model of infraorbital neuroinflammation established by CT-guided injection of talc into the IOF is a simple operation that results in less trauma, stable pain, and a long duration of pain. Moreover, infraorbital neuroinflammation in peripheral branches of the TGN can cause demyelination of the TGN in the intracranial segment.

Background: Autoimmune diseases are associated with cardiovascular and cerebrovascular diseases. However, whether myasthenia gravis (MG) and ischemic stroke (IS) are causally related remains unclear. Objectives: This study aimed to evaluate potential causal links between MG and IS using bidirectional Mendelian randomization (MR). Methods: We conducted a two-sample MR analysis to assess the potential associations between MG and IS. Genetic variants associated with MG and IS as well as their subtypes were extracted from genome-wide association studies by meta-analysis. The inverse-variance weighted method was used for the main MR analysis. Sensitivity analyses, including the MREgger, simple mode, simple median, weighted mode, and weighted median approaches were applied to test the robustness of the results. Results: The MR analyses indicated no causal effects of general MG on IS of all causes (odds ratio [OR] = 0.990, 95% confidence interval [CI]: 0.953-1.029, p = 0.615), large vessel atherosclerosis stroke (OR = 0.943, 95% CI: 0.856-1.039, p = 0.233), cardioembolic stroke (OR = 0.975, 95% CI: 0.867-1.096, p = 0.670), and small vessel occlusion stroke (OR = 1.059, 95% CI 0.974-1.150, p = 0.178). Subgroup analyses indicated no causal effects of early- or late-onset MG on IS and its subtypes (all p > 0.05). The reverse MR analysis showed no significant causal associations of IS on MG (all p > 0.05). Conclusion: Bidirectional MR analysis did not provide evidence to support a causal relationship between genetically predicted MG and IS, although observational studies have found such a potential link.

Background: Previous studies revealed that obstructive sleep apnea (OSA) and smoking, alcohol consumption, and coffee intake are closely related. This study aimed to evaluate the causal effect between these factors and OSA. Methods: The published genome-wide association study data (GWAS) provided genetic tools. We conducted a univariable two-sample Mendelian Randomization (MR) to estimate the causal effect between smoking initiation, never smoking, alcohol consumption, coffee intake, and coffee consumption with the risk of incidence OSA. Inverse variance weighting (IVW) was used as the main method for effect evaluation, and other MR methods were used for sensitivity analysis. After adjusting for body mass index (BMI), hypertension, and diabetes respectively by multivariable MR (MVMR), we further evaluate the causal effect of these factors on OSA. Results: Under univariable MR analysis, we observed that smoking initiation was associated with an increased risk of incidence OSA (OR 1.326, 95% CI 1.001-1.757, p =0.049). Never smoking was associated with decreased risk of OSA (OR 0.872, 95% CI 0.807-0.942, p <0.001). Coffee intake and coffee consumption was associated with an increased incidence of OSA (OR 1.405, 95% CI 1.065-1.854, p =0.016) and (OR 1.330, 95% CI 1.013-1.746, p =0.040). Further multivariate MR showed that the causal relationship between never smoking and OSA existed but not coffee consumption, after adjusting for diabetes and hypertension. However, the all results did not support causality after adjusting for BMI. Conclusion: This two-sample MR study showed that genetically predicted smoking and higher coffee intake are causally associated with an increased risk of OSA.