A matrix metalloproteinase-responsive hydrogel system controls angiogenic peptide release for repair of cerebral ischemia/reperfusion injury

Vascular endothelial growth factor and its mimic peptide KLTWQELYQLKYKGI(QK)are widely used as the most potent angiogenic factors for the treatment of multiple ischemic diseases.However,conventional topical drug delivery often results in a burst release of the drug,leading to transient retention(inefficacy)and undesirable diffusion(toxicity)in vivo.Therefore,a drug delivery system that responds to changes in the microenvironment of tissue regeneration and controls vascular endothelial growth factor release is crucial to improve the treatment of ischemic stroke.Matrix metalloproteinase-2(MMP-2)is gradually upregulated after cerebral ischemia.Herein,vascular endothelial growth factor mimic peptide QK was self-assembled with MMP-2-cleaved peptide PLGLAG(TIMP)and customizable peptide amphiphilic(PA)molecules to construct nanofiber hydrogel PA-TIMP-QK.PA-TIMP-QK was found to control the delivery of QK by MMP-2 upregulation after cerebral ischemia/reperfusion and had a similar biological activity with vascular endothelial growth factor in vitro.The results indicated that PA-TIMP-QK promoted neuronal survival,restored local blood circulation,reduced blood-brain barrier permeability,and restored motor function.These findings suggest that the self-assembling nanofiber hydrogel PA-TIMP-QK may provide an intelligent drug delivery system that responds to the microenvironment and promotes regeneration and repair after cerebral ischemia/reperfusion injury.

Edaravone-loaded poly(amino acid) nanogel inhibits ferroptosis for neuroprotection in cerebral ischemia injury

Neurological injury caused by ischemic stroke is a major cause of permanent disability and death. The currently available neuroprotective drugs fail to achieve desired therapeutic efficacy mainly due to short circulation half-life and poor blood−brain barrier (BBB) permeability. For that, an edaravone-loaded pH/glutathione (pH/GSH) dual-responsive poly(amino acid) nanogel (NG/EDA) was developed to improve the neuroprotection of EDA. The nanogel was triggered by acidic and EDA-induced high-level GSH microenvironments, which enabled the selective and sustained release of EDA at the site of ischemic injury. NG/EDA exhibited a uniform sub-spherical morphology with a mean hydrodynamic diameter of 112.3 ± 8.2 nm. NG/EDA efficiently accumulated at the cerebral ischemic injury site of permanent middle cerebral artery occlusion (pMCAO) mice, showing an efficient BBB crossing feature. Notably, NG/EDA with 50 µM EDA significantly increased neuron survival (29.3%) following oxygen and glucose deprivation by inhibiting ferroptosis. In addition, administering NG/EDA for 7 d significantly reduced infarct volume to 22.2% ± 7.2% and decreased neurobehavioral scores from 9.0 ± 0.6 to 2.0 ± 0.8. Such a pH/GSH dual-responsive nanoplatform might provide a unique and promising modality for neuroprotection in ischemic stroke and other central nervous system diseases.

New Developments in Drug Therapy and Research of Cerebral Vasospasm

In this manuscript a comprehensive coverage of recent developments in the drug therapy of vasospasm while providing the background information that neuroscientists need to understand its rationale. The range of new agents available for treatment of cerebral vasospasm is expanding rapidly along with rapid advances in pharmacology and physiology that are uncovering the mechanisms of this disease. Although there are many publications for treatment of cerebral vaso-spasm, most are focusing on different aspects of vasospasm treatment and many have limited value due to insufficient quality. Moreover, the complexity of this, in many cases deleterious condition, is enormous and the information needed to understand drug effects is accordingly often not readily available in a single source. A number of pharmacological and medical therapies are currently in use or being investigated in an attempt to reverse cerebral vasospasm, but only a few have proven to be useful. Current research efforts promise the eventual production of new medical therapies. At last, recommendations for the use of different treatment stages based on currently available clinical data are provided.

Cerebral ischemia and neuroregeneration

Cerebral ischemia is one of the leading causes of morbidity and mortality worldwide. Although stroke （a form of cerebral ischemia）-related costs are expected to reach 240.67 billion dollars by 2030, options for treatment against cerebral ischemia/stroke are limited. All therapies except anti-thrombolytics （i.e., tissue plasminogen activator） and hypothermia have failed to reduce neuronal injury, neurological deficits, and mortality rates following cerebral ischemia, which suggests that development of novel therapies again st stroke/cerebral ischemia are urgently needed. Here, we discuss the possible mechanism（s） underlying cerebral ischemia-induced brain injury, as well as current and future novel therapies （i.e., growth factors, nicotinamide adenine dinucleotide, melatonin, resveratrol, protein kinase C isozymes, pifithrin, hypothermia, fatty acids, sympathoplegic drugs, and stem cells） as it relates to cerebral ischemia.

Comparison of the anti-apoptotic effects of 15-and 35-minute suspended moxibustion after focal cerebral ischemia/reperfusion injury

Heat-sensitive suspended moxibustion has a neuroprotective effect against focal cerebral ischemia/reperfusion injury, but the underly- ing mechanisms remain unclear. The duration of heat-sensitive suspended moxibustion （usually from 30 minutes to 1 hour） is longer than traditional suspended moxibustion （usually 15 minutes）. However, the effects of 15- and 35-minute suspended moxibustion in rats with cerebra/ischemia/reperfusion injury are poorly understood. In this study, we performed 15- or 35-minute suspended moxibustion at acupoint Dazhui （GV14） in an adult rat model of focal cerebral ischemia/reperfusion injury. Infarct volume was evaluated with the 2,3,5-triphenyltetrazolium chloride assay. Histopathological changes and neuronal apoptosis at the injury site were assessed by hematoxy- lin-eosin staining and terminal deoxynucleotidyl transferase dUTP nick end labeling assay. Caspase-9 and caspase-3 expression at the in- jury site was detected using immunofluorescent staining. Bax and Bcl-2 expression at the injury site was assessed using western blot assay. In the 35-minute moxibustion group, infarct volume was decreased, neuronal apoptosis was reduced, caspase-9, caspase-3 and Bax expres- sion was lower, and Bcl-2 expression was increased, compared with the 15-minute moxibustion group. Our findings show that 35-minute moxibustion has a greater anti-apoptotic effect than 15-minute moxibustion after focal cerebral ischemia/reperfusion injury.

12 hours after cerebral ischemia is the optimal time for bone marrow mesenchymal stem cell transplantation

Cell therapy using stem cell transplantation against cerebral ischemia has been reported. However, it remains controversial regarding the optimal time for cell transplantation and the transplantation route. Rat models of cerebral ischemia were established by occlusion of the middle cerebral artery. At 1, 12 hours, 1, 3, 5 and 7 days after cerebral ischemia, bone marrow mesenchymal stem cells were injected via the tail vein. At 28 days after cerebral ischemia, rat neurological function was evaluated using a 6-point grading scale and the pathological change of ischemic cerebral tissue was observed by hematoxylin-eosin staining. Under the fluorescence microscope, the migration of bone marrow mesenchymal stem cells was examined by PKH labeling. Caspase-3 activity was measured using spectrophotometry. The optimal neurological function recovery, lowest degree of ischemic cerebral damage, greatest number of bone marrow mesenchymal stem cells migrating to peri-ischemic area, and lowest caspase-3 activity in the ischemic cerebral tissue were observed in rats that underwent bone marrow mesenchymal stem cell transplantation at 12 hours after cerebral ischemia. These findings suggest that 12 hours after cerebral ischemia is the optimal time for tail vein injection of bone marrow mesenchymal stem cell transplantation against cerebral ischemia, and the strongest neuroprotective effect of this cell therapy appears at this time.

Exogenous neural stem cell transplantation for cerebral ischemia

Cerebral ischemic injury is the main manifestation of stroke,and its incidence in stroke patients is 70–80%.Although ischemic stroke can be treated with tissue-type plasminogen activator,its time window of effectiveness is narrow.Therefore,the incidence of paralysis,hypoesthesia,aphasia,dysphagia,and cognitive impairment caused by cerebral ischemia is high.Nerve tissue regeneration can promote the recovery of the aforementioned dysfunction.Neural stem cells can participate in the reconstruction of the damaged nervous system and promote the recovery of nervous function during self-repair of damaged brain tissue.Neural stem cell transplantation for ischemic stroke has been a hot topic for more than 10 years.This review discusses the treatment of ischemic stroke with neural stem cells,as well as the mechanisms of their involvement in stroke treatment.

Houshiheisan compound prescription protects neurovascular units after cerebral ischemia

Houshiheisan is composed of wind-dispelling （chrysanthemun fower, divaricate saposhnikovia root, Manchurian wild ginger, cassia twig, Szechwan lovage rhizome, and platycodon root） and deficiency-nourishing （ginseng, Chinese angelica, large-head atractylodes rhizome, Indian bread, and zingiber） drugs. In this study, we assumed these drugs have protective effects against cerebral ischemia, on neurovascular units. Houshiheisan was intragastrically administered in a rat model of focal cerebral ischemia. Hematoxylin-eosin staining, transmission electron microscopy, immu- nofluorescence staining, and western blot assays showed that Houshiheisan reduced pathological injury to the ischemic penumbra, protected neurovascular units, visibly up-regtflated neuronal nuclear antigen expression, and down-regulated amyloid precursor protein and amyloid-[3 42 expression. Wind-dispelling and deficiency-nourishing drugs maintained NeuN expression to varying degrees, but did not affect amyloid precursor protein or amyloid-~ 42 expression in the ischemic penumbra. Our results suggest that the compound prescription Houshiheisan effectively suppresses abnormal amyloid precursor protein accumulation, reduces amyloid substance depo- sition, maintains stabilization of the internal environment of neurovascular units, and minimizes injury to neurovascular units in the ischemic penumbra.

Electroacupuncture effect on synaptic ultrastructure in focal cerebral ischemia marginal zone of the rat

BACKGROUND：Synapses undergo high levels of plasticity within the nervous system, and cerebral ischemia induces synaptic plasticity changes.OBJECTIVE：To demonstrate the effects of electroacupuncture on ultrastructural synaptic changes in the focal cerebral ischemia marginal zone in rats using quantitative analysis of stereological measurement.DESIGN, TIME AND SETTING：A randomized, controlled, animal experiment was performed at the Experimental Animal Center and Laboratory of Electron Microscopy, Guangzhou University of Traditional Chinese Medicine from January 2008 to January 2009.MATERIALS：The G-6805 electric acupuncture apparatus was provided by Shanghai Huayi Instrument Factory, China.METHODS：A total of 90 male, Wistar rats were randomly assigned to sham-surgery, model, and electroacupuncture groups, with 30 animals in each group. Each group was subdivided into 1 hour, as well as 1, 3, 7, and 21 days post-surgery groups, with six animals assigned to each time point. Heat coagulation-induced occlusion of the middle cerebral artery was performed to establish a model of focal cerebral ischemia. Electroacupuncture was applied immediately following surgery to the electroacupuncture group [4/20 Hz, 2.0-3.0 V, 1-3 mA, to Baihui （GV 20） and Dazhui （GV 14）] for 30 minutes. Treatment was performed once a day, and experimental animals were sacrificed, at 1 hour, as well as 1, 3, 7 and 21 days post-surgery.MAIN OUTCOME MEASURES：At different time points after intervention, changes in synaptic ultrastructure, such as postsynaptic density thickness, synaptic cleft width, and synaptic interface curvature, were observed in the focal cerebral ischemia marginal zone in rats through the use of transmission electronic microscopy.RESULTS：Broken synapses were observed following cerebral ischemia, and the number of synapses was significantly decreased. Compared to the model group, synaptic ultrastructure was significantly improved in the electroacupuncture group. Compared to the sham-surgery group, postsynaptic density thickness was significantly decreased, as were synaptic cleft width and synaptic interface curvature in the electroacupuncture and model groups. However, compared to the model group, postsynaptic density thickness was significantly increased in the electroacupuncture group at the same time point post-surgery （P 〈 0.05 or P 〈 0.01）. In addition, synaptic cleft width and synaptic interface curvature were significantly increased with the passage of time （P〈 0.05 or P〈 0.01）.CONCLUSION：Electroacupuncture significantly ameliorated structural synapse lesion during the early stage of cerebral ischemic injury, promoted repair of synaptic structure, improved structural parameters of synapses, and increased synaptic structural plasticity, which suggested that the therapeutic effect of electroacupuncture was related to synaptic reorganization.

Therapeutic potential of bone marrow-derived mononuclear cells for experimental cerebral ischemia in mice

Bone marrow mononuclear cells （BMMCs） can be directly harvested from the donor, allowing for easier application compared with bone marrow mesenchymal stem cells. The present study hypothesized that BMMC transplantation could ameliorate cerebral ischemia in a mouse model. BMMCs were double-labeled with PKH26 and 4＇, 6-diamidino-2-phenylindole （DAPI）, followed by infusion into mice via the tail vein to induce focal cerebral ischemia. At 14 days after transplantation, morphological and neurofunctional recovery were analyzed. PKH26 and DAPI double-positive BMMCs were detected in the cerebral hemisphere of all transplantation mice. Following BMMC administration, there was significant difference in neurofunctional recovery, but no significant difference in survival rates between BMMC-treated mice and other mice. These results demonstrate that transplanted BMMCs migrate to brain tissue and promote neurological function recovery in a mouse model of cerebral ischemia.

Effects of Electroacupuncture at the Conception Vessel on Proliferation and Differentiation of Nerve Stem Cells in the Inferior Zone of the Lateral Ventricle in Cerebral Ischemia Rats

Objective： To observe the effects of electroacupuncture （EA） at the Conception Vessel on proliferation and differentiation of the nerve stem cells in the inferior zone of the lateral ventricle in cerebral ischemia rats. Methods： The model rats were prepared by occlusion of the middle cerebral artery for 2 hours and then by reperfusion. They were randomly divided into two groups： a control group and an EA group. Changes in differentiation and proliferation of the nerve stem cells were observed 7, 14 and 28 days after successful modeling. Results： As compared with the 7-day control group （C-7d group）, there was no significant difference （P〉0.05） in the numbers of 5-bromodeoxyuridine （Brdu） positive cells, Brdu/GFAP, Brdu/Nestin and Brdu/Nse double-labeled cells in the inferior zone of the lateral ventricle in the EA group 7 days after modeling. However, in the 14-day EA group （R-14d group） and the 28-day EA group （R-28d group）, the numbers of Brdu positive cells and Brdu/GFAE Brdu/Nestin, Brdu/Nse double-labeled cells significantly increased as compared respectively with the 14-day control （C-14d group） and the 28-day control （C-28d） group （P〈0.05 or P〈0.01）. Conclusions： EA at the Conception Vessel promotes differentiation and proliferation of the nerve stem cells in the inferior zone of the lateral ventricle in the cerebral ischemia rats, and may stimulate differentiation of the proliferous nerve stem cells towards the astrocvtes.

Neuronal differentiation of adipose-derived stem cells and their transplantation for cerebral ischemia

OBJECTIVE： To review published data on the biological characteristics, differentiation and applications of adipose-derived stem cells in ischemic diseases. DATA RETRIEVAL： A computer-based online search of reports published from January 2005 to June 2012 related to the development of adipose-derived stem cells and their transplantation for treatment of cerebral ischemia was performed in Web of Science using the key words ＂adipose-derived stem cells＂, ＂neural-like cells＂, ＂transplantation＂, ＂stroke＂, and ＂cerebral ischemia＂. SELECTION CRITERIA： The documents associated with the development of adipose-derived stem cells and their transplantation for treatment of cerebral ischemia were selected, and those published in the last 3-5 years or in authoritative journals were preferred in the same field. Totally 89 articles were obtained in the initial retrieval, of which 53 were chosen based on the inclusion criteria. MAIN OUTCOME MEASURES： Biological characteristics and induced differentiation of adipose-derived stem cells and cell transplantation for disease treatment as well as the underlying mechanism of clinical application. RESULTS： The advantages of adipose-derived stem cells include their ease of procurement, wide availability, rapid expansion, low tumorigenesis, low immunogenicity, and absence of ethical constraints. Preclinical experiments have demonstrated that transplanted adipose-derived stem cells can improve neurological functions, reduce small regions of cerebral infarction, promote angiogenesis, and express neuron-specific markers. The improvement of neurological functions was demonstrated in experiments using different methods and time courses of adipose-derived stem cell transplantation, but the mechanisms remain unclear. CONCLUSION： Further research into the treatment of ischemic disease by adipose-derived stem cell transplantation is needed to determine their mechanism of action.

Effect of adenovirus-mediated gene transfer of Olig1 on oligodendrocyte differentiation and remyelination in a rat model of focal cerebral ischemia

BACKGROUND： The transcription factor Oligl is required for oligodendrocyte maturation and demyelinated lesion repair, and is a key regulator of myelinogenesis following ischemia. OBJECTIVE： To examine the efficacy of intraventricular injection of a recombinant adenovirus-expressing Oligl gene （Ad5-Oligl-eGFP） on oligodendrocyte maturation and myelin repair following focal cerebral ischemia. DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment was performed at the Department of Neurology, Beijing Friendship Hospital Affiliated to Capital Medical University from January 2007 to March 2008. MATERIALS： Adenovirus and a recombinant adenovirus containing Oiigl gene （Ad5-Oligl） were provided by Vector Gene Technology, China. METHODS： All 50 rats were induced by middle cerebral artery occlusion. A total of 46 rats were successfully induced and were subsequently randomly assigned to a adenovirus （Ad5） group and recombinant adenovirus-expression Oligl gene （Ad5-Oligl） group, with 23 rats per group. One day after middle cerebral artery occlusion, either Ad5-Oligl-eGFP or Ad5-eGFP （10 μL, 2.3 ×10^11/mL） was injected into the lateral ventricle on the ischemic hemisphere. MAIN OUTCOME MEASURES： Adenovirus-mediated Oligl gene expression in vitro and in vivo was measured by reverse transcription-polymerase chain reaction and immunofluorescence, respectively. Myelin basic protein （MBP） levels were evaluated by Western Blot, immunostaining, and electron microscopy. RESULTS： Exogenous Oligl expression was measured at the periventricular zone of the lateral ventricle 1 day after Ad5-Oligl injection. In the Ad5-Oligl-treated group, MBP protein levels and average intensity of MBP-immunoreactivity （-ir） increased 28 days after middle cerebral artery occlusion, compared with the control group （P 〈 0.01, P 〈 0.05）. Furthermore, myelinated axonal numbers markedly increased following Ad5-Oligl treatment. CONCLUSION： The present data suggested that Ad5-Oligl gene therapy increased MBP expression and the number of remyelinating axons following cerebral ischemia.

Hypertensive-Nimodipine Therapy for Middle Cerebral Artery Vasospasm after Resection of Glioblastoma Multiforme: A Case Report and Literature Review

Delayed cerebral ischemia (DCI) due to post-brain tumor resection vasospasm is an often unrecognized yet debilitating complication. We present a patient with DCI after the resection of glioblastoma multiforme (GBM). To our knowledge, this is the first report on DCI after GBM resection. A 52-year-old female patient with headache for one month underwent subtotal resection of a left temporal GBM encasing the proximal middle cerebral artery (MCA). She was well during the immediate postoperative period but developed right upper limb dense monoparesis on postoperative day four with computed tomographic angiography confirming left MCA vasospasm. Symptoms were significantly alleviated with weeklong hypertensive therapy and nimodipine administration;however they recurred soon after cessation of treatment. A high index of clinical suspicion is needed for the diagnosis of post-tumor resection DCI. Any new postoperative neurological deficit that cannot be explained by hemorrhage, seizures or infection should be expeditiously investigated by angiography or transcranial Doppler sonography. Prompt initiation of hypertensive and nimodipine therapy can possibly reverse neurological deficit. Treatment should be guided by Doppler, angiographic or perfusion imaging studies and not by clinical improvement alone.

~1H-magnetic resonance spectroscopy screening for animals with acute cerebral infraction suitable forthrombolytic therapy

BACKGROUND： As a non-invasive technique which can provide comprehensive biological information, 1H-magnetic resonance spectroscopy （1H-MRS） may provide valuable reference data for irreversible recovery or reversible changes in ischemic tissue after stroke. OBJECTIVE： To monitor and evaluate the effect of the urokinase thrombolytic therapy after experimental acute cerebral ischemia by 1H-MRS technology and investigate its adaptability. DESIGN： Randomly controlled animal study. SETTINGS： Shenzhen Hospital of Peking University and National Key Laboratory of Pattern and Atom ＆ Molecular Physics, Wuhan Physics and Mathematics Institute, Chinese Academy of Science. MATERIALS： Eleven healthy adult Sprague-Dawley （SD） rats, weighing 260–300 g and of both genders, were supplied by Experimental Animal Center of Tongji Medical Collage, Huazhong University of Science and Technology [SCXK （e） 2004-007]. 4.7T superconducting nuclear magnetic resonance meter was provided by Brucker Company. METHODS： The experiment was carried out in Shenzhen Hospital of Peking University and National Key Laboratory of Pattern and Atom ＆ Molecular Physics, Wuhan Physics and Mathematics Institute, Chinese Academy of Science from August 2003 to December 2005. ① The rats were randomly divided into 30-minute self-thrombo-embolism group （n =6） and 60-minute self-thrombo-embolism group （n =5）. Six rats in 30-minute self-thrombo-embolism group were occluded with clot embolus for 30 minutes and 5 rats in 60-minute self-thrombo-embolism group were occluded for 60 minutes. 10 000 U/kg urokinase was dissolved in 2 mL saline and the operation lasted for 5 minutes. ② 1H-MRS was performed before thrombolysis and at 3 hours and 24 hours after successful embolization. The metabolic changes of N-acetyl-L-aspartic acid （NAA）/phosphocreatine （PCr） ＋ creatine （Cr）, choline phosphate （Cho）/PCr＋Cr and lactic acid （Lac）/PCr＋Cr in the region of interests were analyzed. ③ The T2W image was conducted 24 hours after the thrombolytic therapy with TR=500 ms and TE=25 ms. ④ The subjects were sacrificed immediately after 1H-MRS and the brain tissues were cut into pieces and stained with HE method; in addition, pathological changes were observed under optic microscope. MAIN OUTCOME MEASURES： ① Metabolic changes of NAA/PCr＋Cr, Cho/PCr＋Cr and Lac/PCr＋Cr in the region of interests; ② T2W image at 24 hours after the thrombolysis; ③ pathological observation of brain tissue. RESULTS： Eleven rats were all involved in the final analysis. ① Metabolic changes in the region of interests ： In 30-minute self-thrombo-embolism group, the Lac peak emerged immediately after the embolism, but the ischemic zone decreased 3 hours after the thrombolytic therapy （0.252±0.01, 0.603±0.01, P 〈 0.01）. Lac/（PCr＋Cr） ratio was 0.290±0.01 at 24 hours after thrombolysis, which was higher than that at 3 hours after thrombolysis （P 〈 0.01）. The NAA/ （PCr＋Cr） ratio decreased significantly at 3 hours after the thrombolysis as compared with that before thrombolysis （0.922±0.16, 1.196±0.01, P 〈 0.05）. In 60-minute self-thrombo-embolism group, the Lac/（PCr＋Cr） ratio was higher at 3 hours after thrombolysis than that before thrombolysis （0.846±0.12, 0.601±0.11, P 〈 0.05） and the NAA/（PCr＋Cr） decreased at 3 hours after the embolism. Fluctuation of NAA/ （PCr＋Cr） ranged from 0.68 to 0.75 before thrombolysis and from 0.71 to 0.75 at 3 hours after thrombolysis. ② T2W image： T2W image showed that 2 subjects in 30-minute self-thrombo-embolism group whose Lac/NAA was higher than 0.7 suffered from intracranial hemorrhage. This meant that the subjects with Lac/NAA 〉 0.7 were more likely to suffer from intracranial hemorrhage. ③ Histological and morphological examinations： Optic microscope demonstrated that interspace surrounding nerve cells was widened at ischemic center; neurons were swelling; nucleus was stained lightly; pyknosis and mesenchymal edema were mainly observed in lateral cortex of brow and vertex and in lateralpart of corpus striatum. CONCLUSION： ①Compound parameters in ischemic area before thrombolysis should be regarded as an important predicting marker for thrombolytic therapy, effect evaluation and termination. ② 1H-MRS combining with other imaging technique is a detecting way for screening cases who are suitable for thrombolytic therapy.

Clinical Study on the Needling and Drug Treatment of Acute Cerebral Hemorrhage

Researches in recent years have shown that cellular immune factor plays an important role in the generation and development of cerebral hemorrhage1-3.

Effects of neuroprotectant cocktails on focal cerebral ischemia in rats

AIM:To investigate whether cocktail theraphy combined with of neuroprotectants may have more advantages over single agents in treating focal cerebral ischemic cascade.METHODS:With the use of suture occlusin techique,the right middle cerebral artery in rats was occluded.Tirty minutes later,Frutose 1,6 diphosphate(FDP) (50 mg/kg,n=20), MK 801(1 mg/kg,n=20) and N acetylcystein (NAC)(150 mg/kg,n=20) were singly or combinantly infused intraperitoneally.At the same time the cocktail treated group(n=20)were infused the above agents combinationly and the control group(n=20)were infused normal saline intraperitoneally.6 hours and 24 hours after focal cerebral ischemia the animals were weighted and neurologically assessed on 5 point scale.The animals were killed,brains were stained 2,3,5 triphenyltetrazolium chloride for assessment of the infarct volume and then embedden onto slides with paraffin for morphological assessment and terminal transferase dUTP nick ending labeling(TUNEL )were carried out for apoptosis and immunohistochemistry were carried out to investigate the changes in bcl 2.RESULTS:All Neuroprotectants decreased volume of infarction (P< 0.05,F test).While cocktail treated group showed more distinct decrease than other groups(P< 0.05,F test).FDP treated group did not decrease the apoptosis of the neurons and did not increase the bcl 2 expression as well.MK 801 treated group,NAC treated group and cocktail treated group significantly decreased the apoptosis of the neurons and increased the bcl 2 expression (P< 0.05,F test).With cocktail treated group showing more distinct effect (P< 0.05,F test).CONCLUSION:Cocktail may be more effective than single neuroprotectant in this modle.

严重下肢缺血患者围手术期的合理用药

严重下肢缺(CLI)血患者常合并多种慢性疾病,血运重建围手术期的药物管理是疾病治疗的重要内容,不合理的多重用药直接影响患者的治疗结局。本研究针对CLI患者围手术期常见的用药问题进行探讨,重点关注围手术期血栓的预防与治疗、基础疾病防治与多重用药管理、镇痛方案制定与调整,以期为临床医师和药师给患者制定个体化用药方案提供参考。

从铁死亡途径分析针刺对脑缺血再灌注损伤大鼠脑保护的作用机制

目的观察醒脑开窍针刺法对脑缺血再灌注损伤大鼠神经功能和脑组织病理形态学的影响,检测脑缺血再灌注组织内谷胱甘肽(glutathione,GSH)、丙二醛(malondialdehyde,MDA)和谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)、转铁蛋白受体(transferrin receptor,TfR)和膜铁转运蛋白1(ferroportin 1,Fpn1)含量变化,从铁死亡途径探讨针刺对脑缺血再灌注大鼠的脑保护机制。方法25只清洁级雄性Wistar大鼠随机分为假手术组5只、模型组10只、电针组10只。后两组分别再按针刺干预时间分为24 h、72 h两个亚组,每组5只。采用改进的Zea Longa线栓法制作大脑中动脉缺血再灌注模型。电针组予针刺内关、水沟、三阴交,留针期间患侧内关、三阴交穴连接神经穴位刺激仪。采用Zausinger六分法评定神经功能缺损程度,尼氏染色法观察脑组织病理形态改变,酶联免疫吸附测定法对比各组大鼠GSH、MDA、TfR、Fpn1、GPX4的含量。结果与假手术组比较,造模后模型组及电针组神经功能评分均降低(P<0.01)。与模型组比较,电针干预24 h后神经功能评分升高,72 h后评分进一步升高(P<0.01)。造模24 h后,模型组及电针组尼氏小体数量减少,颜色变浅;72 h后模型组尼氏小体更少,电针组尼氏小体数量有一定的恢复。与假手术组比较,模型组MDA、TfR、Fpn1升高、GSH及GPX4降低(P<0.01)。与模型组比较,电针干预24 h及72 h时后MDA、TfR降低、GSH、GPX4及Fpn1升高(P<0.01,P<0.05)。与针刺24 h对比,针刺72 h后MDA、TfR降低更明显,GSH及Fpn1升高更明显(P<0.01,P<0.05)。结论铁死亡是脑缺血再灌注损伤中细胞死亡的重要方式之一,醒脑开窍针刺法可调节脑缺血再灌注大鼠脑组织铁代谢,提高细胞抗氧化能力,抑制神经细胞铁死亡,其脑保护作用与铁死亡途径密切相关。

电针风池穴对MCAO/R大鼠运动功能及运动皮层缺血周围灶Olig2蛋白的影响

目的探讨电针风池穴治疗脑缺血再灌注损伤的可能机制。方法将40只雄性SPF级SD大鼠随机分为空白组、空白电针组、模型组、模型电针组,每组10只。空白组不进行任何处理,空白电针组采用电针风池穴治疗,模型组进行线栓法大脑中动脉闭塞/再灌注(middle cerebral artery occlusion/reperfusion,MCAO/R)模型制备,模型电针组进行线栓法MCAO/R模型制备并采用电针风池穴治疗。4组在电针前后均进行体质量测量、Cat Walk步态检测,电针治疗结束后进行2,3,5-三苯基氯化四氮唑(2,3,5-Triphenyltetrazolium Chloride,TTC)染色观察脑梗死情况,蛋白免疫印迹(Western blot,WB)、免疫荧光检测少突胶质细胞转录因子2(oligodendrocyte transcription factor 2,Olig2)蛋白表达。结果与空白组比较,模型组大鼠脑梗死面积显著增大(P<0.01),体质量显著下降(P<0.01),四肢摆动速度明显减慢(P<0.05),左前脚掌宽度明显减小(P<0.05),WB及免疫荧光检测运动皮层区Olig2表达减少(P<0.05,P<0.01)。与模型组比较,模型电针组大鼠脑梗死面积明显减小(P<0.01),体质量增加(P<0.05),造模后2~3 d大鼠四肢摆动速度增加(P<0.05),造模后3~4 d大鼠左前脚掌宽度增大(P<0.05),WB及免疫荧光检测运动皮层缺血周围灶区域Olig2表达增加(P<0.01)。结论电针风池穴可上调再灌注后大鼠运动皮层区域缺血周围灶Olig2蛋白的表达,减小再灌注后大鼠脑组织梗死面积,促进运动皮层的修复,加速患肢平衡与稳定性的恢复。