OBJECTIVE To investigate the inhibitory effect and mechanism of lead(Pb2+)on γ-amino-butyric acid(GABA)A receptor-mediated currents(IGABA)and GABAergic synaptic transmission in rat cortical neurons.METHODS ①The cortical neurons from 0 d Sprague Dawley(SD)rats were cultured for experiments.The cultured cells(7-14 d)were recorded using the patch-clamp technique to analyze the effects of Pb2+ at different concentrations(1,5,10,50 and 100 μmol·L-1)on IGABA induced by GABA 100 μmol·L-1.② The effects of Pb2+ 50 μmol·L-1 on IGABA induced by GABA at different concentrations(1,10,50,100,500 and 100 μmol·L-1)were detected.③Brain slices(350 μm)were prepared from SD rats(15-19 d).The spontaneous inhibitory post-synaptic currents(sIPSCs),miniature inhibitory post-synaptic currents(mIPSCs)and current injection-induced action potential(AP)were recorded to detect the effects of Pb2+ 10 μmol·L-1 on the amplitude and frequency of sIPSCs and mIPSCs,and the frequency of AP.RESULTS ①Pb2+ inhibited IGABA in a concentration-dependent manner,and IC50 was(68±20)μmol·L-1.②Pb2+ also suppressed the maximum current induced by GABA(P<0.01),with a significant increase of the GABA′s EC50 from(20±6)μmol·L-1 to(87±39)μmol·L-1,indicating that Pb2+ might inhibit IGABA in a non-competitive mechanism.③Pb2+ 10 μmol·L-1 inhibited the frequency(P<0.01)rather than the ampli-tude of sIPSCs reversibly,but had no effect on eigher the frequency or amplitude of mIPSCs.In addi-tion,Pb2+ decreased the frequency of evoked AP by current injection(P<0.01)and reduced the overall excitability of rat cortical neurons.CONCLUSION Pb2+ can significantly inhibit IGABA in primary cultured neurons.In the brain slice experiment,Pb2+ may affect sIPSCs frequency by inhibiting the AP of cortical neurons,suggesting that there are different intrinsic mechanisms through which Pb2+ inhibits both IGABA in primary cultured neurons and the frequency of sIPSCs in brain slice neurons,which points to the complexity of the mechanism of Pb2+ poisoning.

Alzheimer's disease(AD)is a severe threat to human health and one of the three major causes of human death.Double-stranded RNA-dependent protein kinase(PKR)is an interferon-induced protein kinase involved in innate immunity.In the occurrence and development of AD,PKR is upregulated and continu-ously activated.On the one hand,the activation of PKR triggers an integrated stress response in brain cells.On the other hand,it indirectly upregulates the expression of 3-site amyloid precursor protein cleaving enzyme 1 and facilitates the accumulation of amyloid-β protein(Aβ),which could activate PKR activator to further activate PKR,thus forming a sustained accumulation cycle of Aβ.In addition,PKR can promote Tau phosphorylation,thereby reducing microtubule stability in nerve cells.Inflammation in brain tissue,neurotoxicity resulted from Aβaccumulation,and disruption of microtubule stability led to the progression of AD and the declines of memory and cognitive function.Therefore,PKR is a key molecule in the development and progression of AD.Effective PKR detection can aid in the diagnosis and prediction of AD progression and provide opportunities for clinical treat-ment.The inhibitors targeting PKR are expected to control the activity of PKR,thereby controlling the progression of AD.Therefore,PKR could be a target for the development of therapeutic drugs for AD.

Objective:To investigate the association of plasma epoxyeicosatrienoic acids (EETs) with early neurological deterioration (END), and whether EETs are mediated by EPHX2 gene variants in patients with minor ischemic stroke (MIS).Methods:This is a prospective, multi-center observational study in patients with acute MIS in the Chinese population. Acute MIS patients with the first onset and onset within 24 hours who were admitted to Deyang People′s Hospital, the Second Affiliated Hospital of Wenzhou Medical University and the Third Affiliated Hospital of Wenzhou Medical University from March 2013 to June 2015 were recruited. Plasma EETs levels were measured on admission. Single nucleotide polymorphisms of EPHX2 gene rs751141 were genotyped using mass spectrometry. The primary outcome was END within 10 days after admission. END was defined as an increase in National Institutes of Health Stroke Scale score of 2 or more points.Results:A total of 322 patients were enrolled, of which 85 (26.4%) patients experienced END. EETs levels were significantly lower in patients with END [(60.3±7.3) nmol/L] compared to patients without END [(68.4±8.1) nmol/L , t=8.464, P<0.001]. Frequency of EPHX2 gene rs751141 GG was higher in patients with END [66/85(77.6%)] than in patients without END [123/237(51.9%),χ2=17.130, P<0.001], and patients with EPHX2 gene rs751141 GG genotype showed lower EETs levels [GG: (59.6±7.8) nmol/L, AG:(67.9±8.2) nmol/L, AA:(68.8±3.2) nmol/L, F=9.285, P<0.001]. Low level (≤64.3 nmol/L) of EETs was an independent predictor of END (31.5-51.3 nmol/L group: OR=2.96,95% CI 1.18-8.77, P=0.02; 51.4-64.3 nmol/L group: OR=2.46,95% CI 1.06-6.89, P=0.03) in multivariate analyses. END was associated with a higher risk of poor outcome (modified Rankin Scale scores 3-6) at 3 months ( OR=1.82,95% CI 1.46-2.35, P=0.02). Conclusion:END is fairly common and associated with poor outcomes in acute MIS. EPHX2 gene variants may mediate EETs levels, and low levels of EETs are related to END in acute MIS.

Objective:To examine the association of carotid plaque with variants in genes involved in inflammation and endothelial function.Methods:This was a multi-center, cross sectional survey in southwestern China. The residents aged ≥40 years and lived in the community for more than six months volunteered to participate in face-to-face survey in eight communities. A total of 2 377 subjects with high stroke risk were enrolled. Carotid plaque and plaque phenotype were assessed by carotid ultrasound. Genotypes of 19 variants in 10 genes related to inflammation and endothelial function were examined. Gene-gene interaction was analyzed by generalized multifactor dimensionality reduction (GMDR).Results:Carotid plaques were found in 852 (35.8%) subjects, of whom 454 (53.3%) had stable plaques, 398 (46.7%) had vulnerable plaques. PPARA rs4253655 ( OR=1.01, 95% CI 1.03-1.82), HABP2rs7923349 ( OR=1.18, 95% CI 1.06-3.11) and IL1A rs1609682 ( OR=1.09, 95% CI1.03-2.87) were associated with the carotid plaque presence, and NOS2Ars2297518 ( OR=1.05, 95% CI 1.02-2.64) and PPARArs4253655 ( OR=1.00, 95% CI 1.01-1.74) were associated with vulnerable plaque in univariate analysis. GMDR analysis showed that there was a significant gene-gene interaction among HABP2rs7923349, ITGA2rs1991013, IL1Ars1609682 and NOS2Ars8081248, and the high-risk interactive genotype among the four variants was independently associated with a higher risk for carotid vulnerable plaque after adjusting the covariates ( OR=2.81, 95% CI 1.32-7.49, P=0.005). Conclusions:Prevalence of carotid plaque was very high in the high risk stroke population in southwestern China. Variants in genes involved in endothelial function and inflammation were associated with the carotid plaque. The high-risk interactive genotype among HABP2rs7923349, ITGA2rs1991013, IL1Ars1609682 and NOS2Ars8081248 was independently associated with a higher risk for vulnerable plaque.

Objective:To explore the predictive value of regional leptomeningeal collateral circulation scale (rLMC) based on multimodal CT in recanalization of blood vessels in patients with acute ischemic stroke after thrombectomy.Methods:A retrospective analysis was conducted on clinical data of patients with acute ischemic stroke within 6 h of first onset, admitted to our hospital from October 2017 to December 2019. Before operation, the conditions of their vessels were evaluated by rLMC based on multimodal CT. Two areas, anterior cerebral artery (ACA)-middle cerebral artery (MCA) area and posterior cerebral artery (PCA)-MCA area, were divided. The total rLMC scores of two areas (0-10) were calculated: scores of 0-3, scores of 4-7, scores of 8-10. After admission, the recanalization of the blood vessels after thrombectomy was evaluated immediately according to grading of thrombolysis in cerebral infarction (TICI) after completion of thrombectomy within the time window; TICI grading≥II was defined as succeed recanalization. The correlation between rLMC scores and vascular recanalization in patients with acute ischemic stroke was evaluated.Results:Among the 80 patients, 17 were in the rLMC scores of 0-3 group, 25 in the group of rLMC scores of 4-7, and 38 in the group of rLMC scores of 8-10; 68 patients (85.00%) had vascular recanalization, and the success rate of vascular recanalization in patients from the group of rLMC scores of 8-10 was significantly higher than that in the group of rLMC scores of 0-3 (97.36% vs. 58.82%, P<0.05). Correlation analysis results showed that the rLMC score was positively correlated with success rate of vascular recanalization ( r s=0.625, P=0.000); whose sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 89.5%, 90.9%, 94.45%, 83.3% and 90.0%, respectively. Conclusion:The rLMC is closely related to the recanalization rate in patients with acute ischemic stroke after thrombectomy; the success rate of recanalization after intravascular treatment is relatively high in patients with rLMC scores of 8-10.

Objective To investigate the effectiveness and safety of intravenous thrombolytic therapy with low-dose alteplase for elderly patients with acute ischemic stroke.Methods The elderly patients with acute ischemic stroke were enrolled prospectively (onset within 4.5 h,aged ≥75 years).They were randomly divided into either a low-dose group (0.6 mg/kg) or a standard-dose group (0.9 mg/kg).The primary outcome was the modified Rankin Scale (mRS) score determined the good outcome (mRS score 0-2) and poor outcome (mRS score ＞2) at 90 d after treatment;the secondary outcome was intracranial hemorrhage and symptomatic intracranial hemorrhage at 24 h after thrombolysis,and death within 90 d.Results A total of 65 elderly patients with acute ischemic stroke were enrolled,including 32 in the low-dose group and 33 in the standard-dose group.There were no significant differences in all baseline clinical data between the two groups.Compared with before treatment,the proportions of patients with the mRS score 0-2 were significantly higher in both groups at 7 d,14 d,and 90 d after treatment,but there were no significant differences in good outcome rate at the same time point after treatment between the two groups.In addition,there were no significant differences in intracranial hemorrhage (36.4％ vs.25.0％;x2 =0.985,P =0.321),symptomatic intracerebral hemorrhage (21.2％ vs.9.4％;x2 =1.749,P =0.186) at 24 h after treatment,and incidence of death within 90 d after treatment (12.1％ vs.6.3％;x2 =0.151,P=0.697)between the two groups.Conclusions The effectiveness and safety of intravenous thrombolytic therapy with low-dose alteplase (0.6 mg/kg) in elderly patients with acute ischemic stroke is equivalent to the standard dose.

Objective To investigate whether metabolic pathway-related gene polymorphisms are associated with arterial plaque stability and their gene-gene interactions increase the risk of cerebral infarctions.Methods Totally 294 patients with atherothrombosis stroke admitted to the Department of Neurology, the Third Affiliated Hospital of Wenzhou Medical University from September 2010 to December 2012 were divided into a carotid vulnerable plaque group ( n=69 ) and a stable plaque group ( n=225 ) according to the results of carotid B-mode ultrasonography.A total of 282 healthy volunteers excluded carotid plaque and stroke were enrolled as well.Genetic polymorphisms of ALOX5AP and CYP3A5, CYP2C9*2, CYP2C9*3 and EPHX2 were genotyped using polymerase chain reaction and mass spectrometry analysis.The SPSS16.0 software was used to compare genotype frequencies and the generalized multifactor dimensionality reduction ( GMDR ) method was applied for gene-gene interaction analyses.Results The results showed that EPHX2 GG genotype might protect against stroke ( OR =0.520, 95% CI 0.288 -0.940, P=0.030).The distribution of CYP3A5 genotypes showed statistically significant differences (χ2 =7.284, P=0.026) between the vulnerable plaque ( AA: 5 cases, AG: 36 cases, GG: 28 cases) and stable plaque ( AA: 26 cases, AG: 77 cases, GG: 122 cases ) groups.Multivariate Logistic regression analysis showed that the GG genotype of CYP3A5 was protective factor for unstable plaques ( OR=0.405, 95%CI 0.178 -0.920, P =0.031 ).Differences in other SNPs did not reach statistical significance between the two groups.The GMDR analysis showed a significant gene-gene interaction between SG13S114 and A6986G, with scores of 10 for cross-validation consistency and 9 for the sign test (P=0.011).The best model for ischemic stroke was found to be SG13S114 AA and A6986G AA.Adjusting for age, hypertension and diabetes, the certain gene-gene interaction predicted a significantly higher risk of cerebral infarction (OR=1.804, 95%CI 1.180-2.759, P=0.006).Conclusions The EPHX2 G860A gene might be linked with the incidence of cerebral infarctions.Only a CYP3A5 gene polymorphism might be associated with carotid plaque instability in patients with stroke.The gene-gene interaction predicts a significantly higher risk of cerebral infarction.There is a 1.804-fold increased risk for ischemic stroke in individuals with these combined genetic factors.

Objective To investigate the correlation between prostacyclin synthase (prostaglandin I2 synthase, PGIS) gene rs5602 single nucleotide polymorphism and ischemia stroke in Chinese Han population. Methods The patients with ischemia stroke and healthy controls in Chinese Han population were enroled. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS) was used to detect the rs5602 polymorphism. Results A total of 297 patients with ischemic stroke (male 177 and female 120) and 291 healthy controls (male 165, female 126) over the same period were enroled. The frequencies of TT genotype (31. 1% vs. 43. 6% ; χ2 = 5. 773, P = 0. 016) and T alele (56. 8% vs. 65. 8% ; χ2 = 5. 793, P = 0. 016) in the male patients with ischemic stroke were significantly lower than those in the male healthy controls. Multivariate logistic regression analysis showed that the rs5602 TT genotype was a protective factor for ischemic stroke in male (odds ratio 0. 260, 95% confidence interval 0. 118-0. 570; P = 0. 001). Conclusions PGIS gene rs5602 polymorphism is associated with ischemic stroke in male in Chinese Han population.

Objective To evaluate the effectiveness of biofeedback in preventing chronic daily headaches. Methods One hundred patients experiencing daily headaches were randomly divided into a biofeedback group ( n=50) and a drug therapy group (n=50). The patients in the drug therapy groupwere administered a predetermined course of medication. Those in the biofeedback group were given 30 minutes of biofeedback therapy twice a week for 8 weeks, followed by 10 months of intensive therapy once a month. The headache frequency, duration of headache at-tacks, days of using acute pain medication and any other adverse events were recorded 3, 6 and 12 months after the treatment. Results The patients in the biofeedback group had significantly less-frequent headaches, shorter headache attacks and fewer days of using acute pain medications. Conclusion Compared to drug therapy, biofeed-back can prevent chronic daily headachesmore safely and effectively.

BACKGROUND AND PURPOSE: Dual antiplatelet therapy (DAT) with clopidogrel and aspirin has been shown to confer greater protection against early neurological deterioration (END) and early recurrent ischemic stroke (ERIS) than aspirin alone in patients who have experienced an acute ischemic stroke. However, few studies have compared the effects of anticoagulation therapy with low-molecular-weight heparin (LMWH), DAT, and aspirin. METHODS: Patients with acute ischemic stroke (n=1,467) were randomized to therapy groups receiving aspirin (200 mg daily for 14 days, followed by 100 mg daily for 6 months), DAT (200 mg of aspirin and 75 mg of clopidogrel daily for 14 days, then 100 mg of aspirin daily for 6 months), or LMWH (4,000 antifactor Xa IU of enoxaparin in 0.4 mL subcutaneously twice daily for 14 days, followed by 100 mg of aspirin daily for 6 months). The effects of these treatment strategies on the incidence of END, ERIS, and deep-vein thrombosis (DVT) were observed for 10-14 days after treatment, and their impacts on a good outcome were evaluated at 6 months. RESULTS: The DAT and LMWH were associated with a more significant reduction of END and ERIS within 14 days compared with aspirin-alone therapy. In addition, LMWH was associated with a significantly lower incidence of DVT within 14 days. At 6 months, DAT or LMWH improved the outcome among patients aged >70 years and those with symptomatic stenosis in the posterior circulation or basilar artery compared with aspirin. CONCLUSIONS: LMWH or DAT may be more effective than aspirin alone for reducing the incidence of END and ERIS within 14 days, and is associated with improved outcomes in elderly patients and those with stenosis in the posterior circulation or basilar artery at 6 months poststroke.
Aged ; Aspirin* ; Basilar Artery ; Constriction, Pathologic ; Enoxaparin ; Heparin, Low-Molecular-Weight* ; Humans ; Incidence ; Stroke* ; Venous Thrombosis