Experimental studies of relationship between phospholipid level and Yangxueqingnao particles in transient cerebral ischemia attack

AIM:To investigate the effect of transie nt ischemia attack(TIA)on phospho-lipid level and the effect of traditi onal Chinese herb -Yangxueqingnao p ar-ticles.METHODS:To establish the animal model of TIA i n mice by injecting Peroxide through caudal vein,and th en feed the mice with Yangxueqingnao particles in different dose.After e voking 4times of TIA and cut off the head the other day,then measure the level of lysophosphatidic acid(LPA),general phospholipid in blood plasma and bra in tissue.RESULTS:Score of the group without treatment was 1.0±0.8,while that of group administered with Yangxueqingnao particles(2g /kg)was 0.5±0.4(P<0.05)LPA level of TIA mice were 6.1±0.4and 2.1±1.3and increased significantly com-pared with that of the control.This c hanges can be reversed by treatment as can be seen the result 4.9±1.6.general phospholipid level of TIA mice in brain tissue decreased significantly com pared with that of the control as the r esult:(2.3±1.1)and(6.6±2.8)μmol /g,respectively.After treatme nt,general phopholipid level can reach(4.3±2.0)μmol /g.CONCLUSION:LPAand general phospholipid levels in bloo d plasma and brain tissue are abnorma l in experimental TIA mice,but can be rev ersed by Yangxueqingnao particles a nd it can be used in treatment.

Recurrent Transient Ischemic Attacks Revealing Cerebral Amyloid Angiopathy: A Comprehensive Case

This case report investigates the manifestation of cerebral amyloid angiopathy (CAA) through recurrent Transient Ischemic Attacks (TIAs) in an 82-year-old patient. Despite initial diagnostic complexities, cerebral angiography-MRI revealed features indicative of CAA. Symptomatic treatment resulted in improvement, but the patient later developed a fatal hematoma. The discussion navigates the intricate therapeutic landscape of repetitive TIAs in the elderly with cardiovascular risk factors, emphasizing the pivotal role of cerebral MRI and meticulous bleeding risk management. The conclusion stresses the importance of incorporating SWI sequences, specifically when suspecting a cardioembolic TIA, as a diagnostic measure to explore and exclude CAA in the differential diagnosis. This case report provides valuable insights into these challenges, highlighting the need to consider CAA in relevant cases.

Neuroprotective effects of meloxicam on transient brain ischemia in rats:the two faces of anti-inflammatory treatments

The inflammato ry response plays an important role in neuroprotection and regeneration after ischemic insult.The use of non-ste roidal anti-inflammatory drugs has been a matter of debate as to whether they have beneficial or detrimental effects.In this context,the effects of the anti-inflammatory agent meloxicam have been scarcely documented after stro ke,but its ability to inhibit both cyclooxygenase isoforms(1 and 2) could be a promising strategy to modulate postischemic inflammation.This study analyzed the effect of meloxicam in a transient focal cerebral ischemia model in rats,measuring its neuroprotective effect after 48 hours and 7 days of reperfusion and the effects of the treatment on the glial scar and regenerative events such as the generation of new progenitors in the subventricular zone and axonal sprouting at the edge of the damaged area.We show that meloxicam’s neuroprotective effects remained after 7 days of reperfusion even if its administration was restricted to the two first days after ischemia.Moreover,meloxicam treatment modulated glial scar reactivity,which matched with an increase in axonal sprouting.However,this treatment decreased the formation of neuronal progenitor cells.This study discusses the dual role of anti-inflammatory treatments after stro ke and encourages the careful analysis of both the neuroprotective and the regenerative effects in preclinical studies.

Transient ischemic attack induced by pulmonary arteriovenous fistula in a child:A case report

BACKGROUND Cerebral ischemic stroke is attributed to paradoxical cerebral embolism.Pulmonary arteriovenous fistula(PAVF)is a rare potential cause of cerebral ischemic stroke,and cerebral ischemic stroke induced by PAVF in children is rare.CASE SUMMARY We report a case of right PAVF that presented as a transient ischemic attack(TIA)in a 13-year-old boy.The patient underwent embolization therapy and remained clinically stable for 2 years after treatment.CONCLUSION TIA induced by PAVF in children is rare,lacks typical clinical manifestations,and should not be ignored.

Diagnostic value of ABCD2 and ABCD3-Ⅰ risk scoring systems in determining one-month risk of stroke in patients with transient ischemic attack: An observational study

Objective:To compare the diagnostic value of the ABCD2 and ABCD3-栺scoring systems in predicting the development of a 30-day neurological event in adult patients with transient ischemic attack(TIA)presenting to the emergency department.Methods:The study was observational and prospective and was conducted in a single center.The diagnostic values of the ABCD2 and ABCD3-栺scores in predicting a neurological event within one month were compared in patients diagnosed with TIA in the emergency department.Results:A statistically significant difference was observed between groups with or without stroke within one month in terms of both the ABCD2 and ABCD3-栺scores(P=0.044 and P=0.029,respectively).There was no statistically significant difference between the patients with and without a recurrent TIA within one month in relation to the ABCD2 score(P=0.934),but a statistically significant difference was found in the ABCD3-栺scores of these groups(P<0.001).Conclusions:Both the ABCD2 and ABCD3-栺scoring systems could predict ischemic stroke within 30 days of TIA,the ABCD3-栺score is more effective than the ABCD2 score in the prediction of TIA recurrence.

Pretreated Oenan the Javanica extract increases anti-inflammatory cytokines, attenuates gliosis, and protects hippocampal neurons following transient global cerebral ischemia in gerbils

Recently,we have reported that Oenanthe javanica extract(OJE)displays strong neuroprotective effect against ischemic damage after transient global cerebral ischemia.However,neuroprotective mechanisms of OJE have not been fully identified.Thus,this study investigated the neuroprotection of OJE in the hippocampal CA1 area and its anti-inflammatory activity in gerbils subjected to 5 minutes of transient global cerebral ischemia.We treated the animals by intragastrical injection of OJE(100 and 200 mg/kg)once daily for 1 week prior to transient global cerebral ischemia.Neuroprotection of OJE was observed by immunohistochemistry for neuronal nuclear antigen and histofluorescence staining for Fluoro-Jade B.Immunohistochemistry of glial fibrillary acidic protein and ionized calcium-binding adapter molecule 1 was done for astrocytosis and microgliosis,respectively.To investigate the neuroprotective mechanisms of OJE,we performed immunohistochemistry of tumor necrosis factor-alpha and interleukin-2 for pro-inflammatory function and interleukin-4 and interleukin-13 for anti-inflammatory function.When we treated the animals by intragastrical administration of 200 mg/kg of OJE,hippocampal CA1 pyramidal neurons were protected from transient global cerebral ischemia and cerebral ischemia-induced gliosis was inhibited in the ischemic hippocampal CA1 area.We also found that interleukin-4 and-13 immunoreactivities were significantly increased in pyramidal neurons of the ischemic CA1 area after OJE pretreatment,and the increased immunoreactivities were sustained in the CA1 pyramidal neurons after transient global cerebral ischemia.However,OJE pretreatment did not increase interleukin-2 and tumor necrosis factor-alpha immunoreactivities in the CA1 pyramidal neurons.Our findings suggest that pretreatment with OJE can protect neurons and attenuate gliosis from transient global cerebral ischemia via increasing expressions of interleukin-4 and-13.The experimental plan of this study was reviewed and approved by the Institutional Animal Care and Use Committee(IACUC)in Kangwon National University(approval No.KW-160802-1)on August 10,2016.

Characterization of astrocytes and microglial cells in the hippocampal CA1 region after transient focal cerebral ischemia in rats treated with Ilexonin A

Ilexonin A is a compound isolated from the root of Ilex pubescens,a traditional Chinese medicine.Ilexonin A has been shown to play a neuroprotective role by regulating the activation of astrocytes and microglia in the peri-infarct area after ischemia.However,the effects of ilexonin A on astrocytes and microglia in the infarct-free region of the hippocampal CA1 region remain unclear.Focal cerebral ischemia models were established by 2-hour occlusion of the middle cerebral artery in rats.Ilexonin A(20,40 or 80 mg/kg)was administered immediately after ischemia/reperfusion.The astrocyte marker glial fibrillary acidic protein,microglia marker Iba-1,neural stem cell marker nestin and inflammation markers were detected by immunohistochemistry and western blot assay.Expression levels of tumor necrosis factor-αand interleukin 1βwere determined by enzyme linked immunosorbent assay in the hippocampal CA1 tissue.Astrocytes were activated immediately in progressively increasing numbers from 1,3,to 7 days post-ischemia/reperfusion.The number of activated astrocytes further increased in the hippocampal CA1 region after treatment with ilexonin A.Microglial cells remained quiescent after ischemia/reperfusion,but became activated after treatment with ilexonin A.Ilexonin A enhanced nestin expression and reduced the expression of tumor necrosis factor-αand interleukin 1βin the hippocampus post-ischemia/reperfusion.The results of the present study suggest that ilexonin A has a neuroprotective effect in the hippocampus after ischemia/reperfusion,probably through regulating astrocytes and microglia activation,promoting neuronal stem cell proliferation and reducing the levels of pro-inflammatory factors.This study was approved by the Animal Ethics Committee of the Fujian Medical University Union Hospital,China.

Glucose metabolism and neurogenesis in the gerbil hippocampus after transient forebrain ischemia

Recent evidence exists that glucose transporter 3（GLUT3） plays an important role in the energy metabolism in the brain.Most previous studies have been conducted using focal or hypoxic ischemia models and have focused on changes in GLUT3 expression based on protein and m RNA levels rather than tissue levels.In the present study,we observed change in GLUT3 immunoreactivity in the adult gerbil hippocampus at various time points after 5 minutes of transient forebrain ischemia.In the sham-operated group,GLUT3 immunoreactivity in the hippocampal CA1 region was weak,in the pyramidal cells of the CA1 region increased in a time-dependent fashion 24 hours after ischemia,and in the hippocampal CA1 region decreased significantly between 2 and 5 days after ischemia,with high level of GLUT3 immunoreactivity observed in the CA1 region 10 days after ischemia.In a double immunofluorescence study using GLUT3 and glial-fibrillary acidic protein（GFAP）,we observed strong GLUT3 immunoreactivity in the astrocytes.GLUT3 immunoreactivity increased after ischemia and peaked 7 days in the dentate gyrus after ischemia/reperfusion.In a double immunofluorescence study using GLUT3 and doublecortin（DCX）,we observed low level of GLUT3 immunoreactivity in the differentiated neuroblasts of the subgranular zone of the dentate gyrus after ischemia.GLUT3 immunoreactivity in the sham-operated group was mainly detected in the subgranular zone of the dentate gyrus.These results suggest that the increase in GLUT3 immunoreactivity may be a compensatory mechanism to modulate glucose level in the hippocampal CA1 region and to promote adult neurogenesis in the dentate gyrus.

Time-course pattern of neuronal loss and gliosis in gerbil hippocampi following mild, severe, or lethal transient global cerebral ischemia

Transient ischemia in the whole brain leads to neuronal loss/death in vulnerable brain regions. The striatum, neocortex and hippocampus selectively loose specific neurons after transient ischemia. Just 5 minutes of transient ischemia can cause pyramidal neuronal death in the hippocampal cornu ammonis (CA) 1 field at 4 days after transient ischemia. In this study, we investigated the effects of 5-minute (mild), 15-minute (severe), and 20-minute (lethal) transient ischemia by bilateral common carotid artery occlusion (BCCAO) on behavioral change and neuronal death and gliosis (astrocytosis and microgliosis) in gerbil hippocampal subregions (CA1-3 region and dentate gyrus). We performed spontaneous motor activity test to evaluate gerbil locomotor activity, cresyl violet staining to detect cellular distribution, neuronal nuclei immunohistochemistry to detect neuronal distribution, and Fluoro-Jade B histofluorescence to evaluate neuronal death. We also conducted immunohistochemical staining for glial fibrillary acidic protein and ionized calcium-binding adapter molecule 1 (Ibal) to evaluate astrocytosis and microgliosis, respectively. Animals subjected to 20-minute BCCAO died in at least 2 days. BCCAO for 15 minutes led to pyramidal cell death in hippocampal CA1-3 region 2 days later and granule cell death in hippocampal de匚tate gyrus 5 days later. Similar results were not found in animals subjected to 5-minute BCCAO. Gliosis was much more rapidly and severely progressed in animals subjected to 15-minute BCCAO than in those subjected to 5- minute BCCAO. Our results indicate that neuronal loss in the hippocampal formation following transient ischemia is significantly different according to regions and severity of transient ischemia. The experimental protocol was approved by Institutional Animal Care and Use Committee (AICUC) of Kangwon National University (approval No. KW-180124-1) on May 22, 2018.

Cerebral perfusion in corresponding blood supply areas of transient ischemic attack patients with intracranial stenosis Seven cases of diamox-perfusion verified by magnetic resonance-perfusion-weighted imaging

BACKGROUND： Due to collateral circulation and cerebrovascular reserve, arterial stenosis and reduced cerebral blood flow may not necessarily indicate impaired cerebral peffusion. Therefore, according to degree of stenosis and clinical symptoms, interventional surgery to relieve arterial stenosis in transient ischemic attack （TIA） patients with major intracranial stenosis is imprudent. Rather, cerebral perfusion and reserve capacity are direct indicators for the assessment of degree and presence of cerebral ischemia. OBJECTIVE： To evaluate cerebral perfusion and reserve in TIA patients with major intracranial stenosis or occlusion using magnetic resonance-perfusion-weighted imaging （MR-PWl） data prior to and following diamox administration. DESIGN, TIME AND SETTING： A self-comparative, neuroimaging observation was performed at the Neurological Department and Radiological Center of the First Affiliated Hospital of Jinan University between December 2007 and April 2009. PARTICIPANTS： Seven acute TIA patients, who were admitted to the Neurological Department of the First Affiliated Hospital of Jinan University between December 2007 and April 2009, were enrolled in the present study. Magnetic resonance imaging confirmed that no acute cerebral infarction happened, nor did bleeding exist. Magnetic resonance angiography, transcranial Doppler ultrasound, and/or digital subtraction angiography confirmed the presence of major intracranial arterial stenosis. Clinical symptoms corresponded to blood supplying regions of the arterial stenosis. METHODS： Baseline MR-PWI was performed on seven patients with intracranial stenosis or occlusion. Two grams of acetazolamide （diamox） were orally administered after 2 days. A second PWl was performed after 2 hours to compare cerebral perfusion parameters prior to and following diamox administration. MAIN OUTCOME MEASURES： PWI results of cerebral perfusion prior to and following diamox administration. RESULTS： The baseline PWl from five patients indicated decreased cerebral perfusion areas. Following oral administration of diamox, cerebral perfusion significantly decreased in those areas. Moreover, new areas of decreased cerebral perfusion were observed in two out of the five patients. In one patient, no significant decrease in cerebral perfusion was found. In another patient, baseline PWl indicated decreased cerebral perfusion in the left hemisphere. However, normal perfusion was observed in both cerebral lobes following diamox administration. CONCLUSION： TIA patients with intracranial stenosis, who are diagnosed by PWI and exhibited decreased cerebral perfusion and reserve, might require further treatment such as intervention by angioptasty.

Awareness intervention for Beijing neurologists regarding secondary prevention of cerebral infarction/transient ischemia Cross-sectional investigation

BACKGROUND： Stroke prevention guidelines should be made available application to aid in uniformity, timing, preciseness, and acceptance of disease. OBJECTIVE： To investigate the awareness of neurologists in some Beijing secondary prevention of cerebral infarction/transient ischemia. DESIGN： Cross-sectional study. to neurologists for clinical hospitals of intervention in SETTING： Beijing Tiantan Hospital, Beijing Anzhen Hospital, General Hospital of Beijing Military Area, Command of Chinese PLA, Beijing Chuiyangliu Hospital, Beijing 6^th Hospital, Beijing Hepingli Hospital, and Beijing Daxing District Hospital. PARTICIPANTS： A total of 28 （associate） chief physicians, 58 attending physicians, and 54 resident physicians who engaged in clinical treatment of cerebrovascular diseases were selected from 8 hospitals in Beijing from March to April 2007. All physicians provided informed consent. METHODS： Self-made closed questionnaires were provided for data collection, consisting of 16 questions that were single choice or multiple choice. Specifically, questions 1-7 focused on awareness of blood pressure regulation in different patients and first choice of decompression drug; questions 8-12 focused on awareness of lipid regulation; and questions 13-16 focused on awareness of anti-blood platelet drugs applied in secondary prevention. The scores ranged from 0-100 points, and each question was worth 6.25 points. The scores positively correlated with the awareness rate. To test leveling real-time, the survey lasted for a maximum of 20 minutes. One questionnaire was independently finished by each subject in the survey. MAIN OUTCOME MEASURES： Awareness intervention among neurologists during secondary prevention of cerebral infarction/transient ischemia and questionnaire scores. RESULTS： 140 subjects were included in the final analysis. ① The awareness rate among neurologists for intervention during secondary prevention of cerebral infarction/transient ischemia ranged from 0.7-57.9%, the scores ranged between 0-56 points, and the mean score was 26 points.② Scores of resident physicians were 0-56 points with a mean score of 26 points; attending physicians scored 6-50 points, with a mean score of 26 points; and chief physicians scored 6-50 points, with a mean score of 23 points. There were no significant differences among the various physicians （F = 0.771, t = 0.465, P 〉 0.05）. CONCLUSION： Awareness among neurologists of intervention during secondary prevention of cerebral infarction/transient ischemia is not ideal. However, there was no significant difference between professional titles.

Yizhi Xingnao prescription improves the cognitive function of patients after a transient ischemic attack

Patients with mild cognitive impairment after a transient ischemic attack were included in this study. They were treated with Yizhi Xingnao prescription, ergoloid mesylates or aspirin for 60 days. Evaluation using the Montreal Cognitive Assessment Scale showed that cognitive function was significantly improved in all patients, especially after the combined treatment of Yizhi Xingnao and aspirin. The scores from the Montreal Cognitive Assessment Scale were improved overall and the effective treatment rate was as high as 79%, which was higher than patients treated with a combination of ergoloid mesylates and aspirin, or aspirin alone. Our experimental findings indicate that YizhiXingnao prescription can improve mild cognitive impairment after a transient ischemic attack, and that it is more effective than ergoloid mesylates.

Clinical analysis on neuroprotection of transient ischemic attacks

Transient ischemic attack（TIA） is an acute cerebrovascular incident,and is generally considered the best opportunity for early neuroprotective treatment against cerebral ischemia.This study retrospectively analyzed 80 patients with TIA（38 males and 42 females）.Among 61 patients who received neuroprotective cerebrolysin treatment within 24 hours after TIA onset,13（21.31%） patients suffered subsequent strokes.Among 19 patients who received neuroprotective cerebrolysin treatment within 24-72 hours after TIA onset,seven（36.84%） developed cerebral infarction.There was a significant difference in the proportion of subsequent strokes between patients receiving cerebrolysin treatment within 24 hours and 24-72 hours after TIA onset（P = 0.438）.These findings suggest that neuroprotective drugs administrated within 24 hours after TIA onset help reduce the incidence of subsequent strokes.The results demonstrate usefulness of the ABCD2 score at TIA patients in the determination of short-term and long-term cerebrovascular risk,including the frequency of subsequent ischemic cerebral infarctions up to 12 months.

Wall shear stress can improve prediction accuracy for transient ischemic attack

BACKGROUND Early prediction of transient ischemic attack (TIA) has important clinical value. To date, systematic studies on clinical, biochemical, and imaging indicators related to carotid atherosclerosis have been carried out to predict the occurrence of TIA. However, their prediction accuracy is limited. AIM To explore the role of combining wall shear stress (WSS) with conventional predictive indicators in improving the accuracy of TIA prediction. METHODS A total of 250 patients with atherosclerosis who underwent carotid ultrasonography at Naval Military Medical University Affiliated Gongli Hospital were recruited. Plaque location, plaque properties, stenosis rate, peak systolic velocity, and end diastolic velocity were measured and recorded. The WSS distribution map of the proximal and distal ends of the plaque shoulder was drawn using the shear stress quantitative analysis software, and the average values of WSS were recorded. The laboratory indicators of the subjects were recorded. The patients were followed for 4 years. Patients with TIA were included in a TIA group and the remaining patients were included in a control group. The clinical data, laboratory indicators, and ultrasound characteristics of the two groups were analyzed. Survival curves were plotted by the Kaplan-Meier method. Receiver operating characteristic curves were established to evaluate the accuracy of potential indicators in predicting TIA. Logistic regression model was used to establish combined prediction, and the accuracy of combined predictive indicators for TIA was explored.RESULTS The intraclass correlation coefficients of the WSS between the proximal and distal ends of the plaque shoulder were 0.976 and 0.993, respectively, which indicated an excellent agreement. At the end of the follow-up, 30 patients suffered TIA (TIA group) and 204 patients did not (control group). Hypertension (P = 0.037), diabetes (P = 0.026), homocysteine (Hcy)(P = 0.022), fasting blood glucose (P = 0.034), plaque properties (P = 0.000), luminal stenosis rate (P = 0.000), and proximal end WSS (P = 0.000) were independent influencing factors for TIA during follow-up. The accuracy of each indicator for predicting TIA individually was not high (area under the curve [AUC]< 0.9). The accuracy of the combined indicator including WSS (AUC = 0.944) was significantly higher than that of the combined indicator without WSS (AUC = 0.856) in predicting TIA (z = 2.177, P = 0.030). The sensitivity and specificity of the combined indicator including WSS were 86.67% and 92.16%, respectively. CONCLUSION WSS at plaque surface combined with hypertension, diabetes, Hcy, blood glucose, plaque properties, and stenosis rate can significantly improve the accuracy of predicting TIA.

Executive function during transient ischemic attacks

BACKGROUND： Recent researches demonstrate that damage of executive function is an early manifestation of vascular cognitive disorder. OBJECTIVE： To investigate the executive functions of patients with transient ischemic attack （TIA）. DESIGN： Case control. SETTING： Department of Neurology, the First Affiliated Hospital of Xinxiang Medical College. PARTICIPANTS： A total of 83 TIA patients （46 males and 37 females, aged 32 - 74 years） were selected from Department of Neurology, the First Affiliated Hospital of Xinxiang Medical College from July 2005 to December 2006. The diagnosis of TIA was established by the criteria of cerebrovascular diseases made by the Fourth National Cerebrovascular Disease Meeting. In these cases, forty-five patients （54%） were involved in internal carotid artery, and thirty-eight patients （46%） were involved in vertebral basilar system. Fifty healthy adults （Control group： 28 males and 22 females, aged 32 - 74 years） were chosen from retiree in community and family of patients. All of them were right-handedness, without cerebrovascular disease, cerebral trauma, inborn oligophrenia, and heating and visual disorders; also they had no anxiety and depression nearly one week. In addition, all the subjects cooperated with examination. METHODS： After TIA diagnosis, all patients received neuropsychological examination, including attention and inhibition, working memory, flexibility, planning and diversion, based on Stroop tests （C and CW） and the Wisconsin card sorting tests （WCST）. ① WCST test： The edition revised by Nelson was used. We would take the times of sorting, incorrect response, persistent incorrect response, and randomly incorrect response as the index of evaluation.② Stroop tests： The edition revised by Trenarry was used and this test had two types： form color and form color-word. Their incorrect response and response time were recorded for data analysis in the end. MAIN OUTCOME MEASURES： Results of WCST test and Stroop tests. RESULTS： A total of 83 TIA patients and 50 healthy subjects were involved in the final analysis. Items of Stroop tests, including response time of Stroop-C test, incorrect response of Stroop-C test, response time of Stroop-CW test and incorrect response of Stroop-CW test, and items of WCST, including incorrect response, persistent incorrect response and random incorrect response, between internal carotid artery system and vertebral basilar artery system were （93.87±24.39） s, 3.16±3.97, （228.46±68.13） s, 12.91 ± 10.56, 55.42± 20.38, 26.58± 10.41, 28.62±11.22; （96.76±24.75） s, 3.89±4.77, （223.46±72.54） s, 18.71 ± 13.80, 52.47 ±22.25, 28.82± 16.20, 23.39±9.72, respectively; this was higher than those in the control group [（52.10± 12.18） s, 1.68± 1.97, （134.86±34.15） s, 4.22±4.21, 32.46± 17.97, 14.42±8.47, 18.24± 10.24, P 〈 0.05 - 0.01]. Times of sorting of WCST between internal carotid artery system and vertebral basilar artery system were obviously lower than those in the control group （8.27±2.73, 8.66±2.77, 11.22±2.41, P 〈 0.01）. However, there were no significant differences between internal carotid artery system and vertebral basilar artery system （P 〉 0.05）. CONCLUSION： Patients with TIA have executive dysfunctions, and the dysfunctions have no difference between internal carotid artery system and vertebral basilar artery system.

Radix Ilicis Pubescentis total flavonoids ameliorates neuronal damage and reduces lesion extent in a mouse model of transient ischemic attack

Flavonoids are a major component in the traditional Chinese medicine Radix Ilicis Pubescentis.Previous studies have shown that the administration of Radix Ilicis Pubescentis total flavonoids is protective in cerebral ischemia.However,to our knowledge,no studies have examined whether the total flavonoids extracted from Radix Ilicis Pubescentis prevent or ameliorate neuronal damage following transient ischemic attacks.Therefore,Radix Ilicis Pubescentis total flavonoids question and the potential underlying mechanisms.Thus,beginning 3 days before the induction of a mouse model of transient ischemic attack using tert-butyl hydroperoxide injections,mice were intragastrically administered 0.3,0.15,or 0.075 g/kg of Radix Ilicis Pubescentis total flavonoids daily for 10 days.The results of spectrophotometric analyses demonstrated that Radix Ilicis Pubescentis total flavonoids enhanced oxygen free radical scavenging and reduced pathological alterations in the brain.Hematoxylin-eosin staining results showed that Radix Ilicis Pubescentis total flavonoids reduced hippocampal neuronal damage and cerebral vascular injury in this mouse model of transient ischemic attack.These results suggest that the antioxidant effects of Radix Ilicis Pubescentis total flavonoids alleviate the damage to brain tissue caused by transient ischemic attack.

Nerve growth factor downregulates c-jun mRNA and Caspase-3 in striate cortex of rats after transient global cerebral ischemia/reperfusion

BACKGROUND： Immediate early gene （lEG） c-jun is a sensitive marker for functional status of nerve cells. Caspase-3 is a cysteine protease, which is a critical regulator of apoptosis. The effect of exogenous nerve growth factor （NGF） on the expression of c-jun mRNA and Caspase-3 protein in striate cortex of rats with transient global cerebral ischemia/reperfusion （IR） is unclear. OBJECTIVE： To study the protective effect of exogenous NGF on the brain of rats with transient globa cerebral IR and its effecting pathway by observing the expression of c-jun mRNA and Caspase-3 protein. DESIGN： Randomized controlled animal trial SETTING： Department of Neural Anatomy, Institute of Brain, China Medical University MATERIALS：Eighteen healthy male SD rats of clean grade, aged 1 to 3 months, with body mass of 250 to 300 g, were involved in this study. NGF was provided by Dalian Svate Pharmaceutical Co.,Ltd. c-jun in situ hybridization detection kit, Caspase-3 antibody and SABC kit were purchased from Boster Biotechnology Co.. Ltd. METHODS： This trial was carried out in the Department of Neural Anatomy, Institute of Brain, China Medical University during September 2003 to April 2005. （1） Experimental animals were randomized into three groups with 6 in each： sham-operation group, IR group and NGF group.（2）After the rats were anesthetized, the bilateral common carotid arteries and right external carotid arteries of rats were bluntly dissected and bilateral common carotid arteries were clamped for 30 minutes with bulldog clamps. Reperfusion began after buldog clamps were removed. Normal saline of lmL and NGF （1×10^6 U/L） of 1 mL was injected into the common carotid artery of rats via right external carotid arteries in the IR group and NGF group respectively. The injection was conducted within 30 minutes, and then the right external carotid arteries were ligated. In the sham-operation group, occlusion of bilateral common carotid arteries and administration of drugs were omitted.GAll the rats were executed by decollation at 3 hours after modeling. The animals were fixed with phosphate buffer solution （PBS, 0.1 mol/L） containing 40 g/L polyformaldehyde, their brains were quickly removed. The coronal section tissue mass containing striate cortex about 3 mm before line between two ears was taken and made into successive frozen sections.（4）The expression of c-jun mRNA and Caspase-3 protein in striate cortex of global cerebral ischemia rats were detected with in situ hybridization, immunohistochemistry and microscope image analysis. （5）t test was used for comparing the difference of the measurement data. MAIN OUTCOME MEASURES：Comparison of the expression of lEG c-jun mRNA and Caspase-3 protein in striate cortex of brain of rats in each group. RESULTS：All the 18 SD rats were involved in the analysis of results. The c-jun mRNA and Caspase-3 protein positive reaction cells were found brown yellow in the striate cortex of rats, and most of them were in lamellas Ⅱ and Ⅲ, mainly presenting round or oval. The expression of c-jun mRNA and Caspase-3 protein in sham-operation group was weak or negative. The average gray value of c-jun mRNA and Caspase-3 protein in the IR group was significantly lower than that in the sham-operation group （49.52±4.13 vs. 95.48± 5.28; 74.73±4.29 vs. 162.38±9.16,P 〈 0.01）. The average gray value of c-jun mRNA and Caspase-3 protein in the NGF group was significantly higher than that in the IR group （63.96±4.25 vs.49.52±4.13; 83.98± 4.13 vs. 74.73±4.29, P〈 0.05）. CONCLUSION： NGF can protect ischemic neurons by down-regulating the expression of c-jun mRNA and Caspase-3 protein in striate cortex of global cerebral ischemia rats.

Aggregation Patterns of Proteasome in Injured Neurons Induced by Transient Cerebral Ischemia

Proteasome activity reduction is an important pathological phenomenon, resulting in proteins aggregation and neuronal death in the injured neurons induced by transient ischemia. Our previous report showed that the trap of proteasome in the protein aggregates was a reason to lead to the reduction of proteasome activity. However, the patterns of proteasome entered into protein aggregates are not clear. In this study, we used a global ischemia model, Hematoxylin-Eosin staining, differential centrifuge, proteasome activity assay, sucrose gradient density centrifuge, and Western blot analysis to investigate this problem. Our results show that there are two aggregation patterns of proteasome after transient ischemia and reperfusion. One is that 26S proteasome is trapped by protein aggregates as a whole unit, and the other is that 19S or 20S is trapped in the protein aggregates, respectively, after 26S disassociates.

Proteasome alteration and delayed neuronal death in hippocampal CA1 and dentate gyrus regions following transient cerebral ischemia

BACKGROUND： Proteasome dysfunction has been reported to induce abnormal protein aggregation and cell death. OBJECTIVE： To investigate the effect of proteasome changes on delayed neuronal death in CA1 and dentate gyrus （DG） regions of the rat hippocampus following transient cerebral ischemia. DESIGN, TIME AND SETTING： A randomized, controlled animal experiment. The study was performed at the Department of Biochemistry and Molecular Biology, Norman Bethune Medical College of Jilin University, from September 2006 to May 2008. MATERIALS： Rabbit anti-19S S10B polyclonal antibody was purchased from Bioreagents, USA; propidium iodide and fluorescently-labeled goat anti-rabbit IgG were purchased from Jackson Immunoresearch, USA; hematoxylin and eosin staining solution was purchased from Sigma, USA; LSM 510 confocal microscope was purchased from Zeiss, Germany. METHODS： A total of 40 healthy Wistar rats, male, 4 months old, were randomly divided into sham surgery group （n = 8） and model group （n = 32）. Ischemic models were established in the model group by transient clamping of the bilateral carotid arteries and decreased blood pressure. After 20 minutes of global ischemia, the clamp was removed to allow blood flow for 30 minutes, 4, 24 and 72 hours, respectively, with 8 rats at each time point. The bilateral carotid arteries were not ligated in the sham surgery group. MAIN OUTCOME MEASURES： Neuronal death in the CA1 and DG regions was observed by hematoxylin-eosin staining. Proteasome expression in CA1 and DG region neurons was detected by immunohistochemistry. RESULTS： Hematoxylin-eosin staining showed neuronal death in the CA1 region alone at 72 hours of reperfusion following ischemia. In comparison to the sham surgery group, a significant decrease in proteasome expression was observed, by immunohistochemistry, in the CA1 and DG regions in the model group, following 30 minutes, 4, 24, and 72 hours of reperfusion （P 〈 0.01）. After 72 hours of reperfusion following ischemia, proteasome expression had almost completely disappeared in the CA1 region. In contrast, neurons of the DG region showed minimized proteasome expression at 24 hours, with a slight increase at 72 hours （P 〈 0.01）. CONCLUSION： The alteration of proteasome following ischemia/reperfusion in the neurons of hippocampal CA1 and DG regions reduces the ability of cells to degrade abnormal protein, which may be an important factor resulting in delayed neuronal death following transient cerebral ischemia.

Local cerebral blood perfusion correlates with nerve fibre integrity in transient ischemic attack patients with middle cerebral artery stenosis: a pilot study

Recent advances in neuroimaging contribute a lot to the accurate diagnosis and evaluation of cerebrovascular diseases. To explore the relationship among blood perfusion, metabolism and brain structure integrity, 6 Chinese transient ischemic attack (TIA) patients with middle cerebral artery (MCA) stenosis were examined by xenon-enhanced computed tomography (Xe-CT), magnetic resonance spectroscopy (MRS) and diffusion tensor imaging (DTI) to compare cerebral blood flow (CBF) values, (choline + creatine)/ N-acetyl aspartate [(Cho + Cr)/NAA] values and fractional anisotropy (FA) values in the MCA territory. Our results showed that CBF values significantly decreased in the ipsilateral basal ganglion regions in all 5 cases with unilateral MCA stenosis, with a corresponding decrease of FA values in the same region. In conclusion, decreased blood perfusion may indicate nerve fibre damage in the dominating regions of stenosed arteries.