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1.
局灶性脑缺血耐受和星形胶质细胞反应   总被引:11,自引:1,他引:11  
目的 研究短暂性局灶性脑缺血预处理对永久性局灶性脑缺血的保护作用 ,及最佳预处理时间剂量 ,并探讨星形胶质细胞在脑缺血耐受中的反应。方法 采用开颅方法阻断大鼠大脑中动脉 ,通过观察大鼠脑梗死后神经功能损伤状况、脑梗死体积分析及病理形态学变化 ,评价不同的缺血预处理时间剂量 (10分钟、2 0分钟、30分钟 )对永久性局灶性脑缺血的保护作用。采用胶质纤维酸性蛋白 (GFAP)免疫组化法观察星形胶质细胞在脑缺血耐受中的反应。结果 与对照组相比 ,缺血预处理 2 0分钟未引起明显的神经元损伤 ,但使永久性局灶性脑缺血后神经功能损伤减轻 ,梗死体积明显减小 (P <0 .0 1)。免疫组化显示 ,2 0分钟缺血预处理组及重复缺血组星形胶质细胞在损伤预处理侧广泛激活。结论  2 0分钟局灶性脑缺血预处理能够有效诱导脑缺血耐受。星形胶质细胞的激活可能与脑缺血耐受中神经元的存活相关。  相似文献   

2.
BACKGROUND: Blood supply to the hippocampus is not provided by the middle cerebral artery. However, previous studies have shown that delayed neuronal death in the hippocampus may occur following focal cerebral ischemia induced by middle cerebral artery occlusion.
OBJECTIVE: To observe the relationship between reactive changes in hippocampal astrocytes and delayed neuronal death in the hippocampal CA1 region following middle cerebral artery occlusion.
DESIGN, TIME AND SETTING: The immunohistochemical, randomized, controlled animal study was performed at the Laboratory of Department of Neurology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, from July to November 2007.
MATERIALS: Rabbit anti-glial fibrillary acidic protein (GFAP) (Neomarkers, USA), goat anti-rabbit IgG (Sigma, USA) and ApoAlert apoptosis detection kit (Biosciences Clontech, USA) were used in this study. METHODS: A total of 42 healthy adult male Wistar rats, aged 3–5 months, were randomly divided into a sham operation group (n = 6) and a cerebral ischemia/reperfusion group (n = 36). In the cerebral ischemia/reperfusion group, cerebral ischemia/reperfusion models were created by middle cerebral artery occlusion. In the sham operation group, the thread was only inserted into the initial region of the internal carotid artery, and middle cerebral artery occlusion was not induced. Rats in the cerebral ischemia/reperfusion group were assigned to a delayed neuronal death (+) subgroup and a delayed neuronal death (–) subgroup, according to the occurrence of delayed neuronal death in the ischemic side of the hippocampal CA1 region following cerebral ischemia.
MAIN OUTCOME MEASURES: Delayed neuronal death in the hippocampal CA1 region was measured by Nissl staining. GFAP expression and delayed neuronal death changes were measured in the rat hippocampal CA1 region at the ischemic hemisphere by double staining for GFAP and TUNEL.
RESULTS: After 3 days of ischemia/reperfusion, astrocytes with abnormal morphology were detected in the rat hippocampal CA1 region in the delayed neuronal death (+) subgroup. No significant difference in GFAP expression was found in the rat hippocampal CA1 region at the ischemic hemisphere in the sham operation group, delayed neuronal death (+) subgroup and delayed neuronal death (–) subgroup (P 〉 0.05). After 7 days of ischemia/reperfusion, many GFAP-positive cells, which possessed a large cell body and an increased number of processes, were activated in the rat hippocampal CA1 region at the ischemic hemisphere. GFAP expression in the hippocampal CA1 region was greater in the delayed neuronal death (+) subgroup and delayed neuronal death (–) subgroup compared with the sham operation group (P 〈 0.01). Moreover, GFAP expression was significantly greater in the delayed neuronal death (–) subgroup than in the delayed neuronal death (+) subgroup (P 〈 0.01). After 30 days of ischemia/reperfusion, GFAP-positive cells were present in scar-like structures in the rat hippocampal CA1 region at the ischemic hemisphere. GFAP expression was significantly greater in the delayed neuronal death (+) subgroup and delayed neuronal death (–) subgroup compared with the sham operation group (P 〈 0.05). GFAP expression was significantly lower in the delayed neuronal death (–) subgroup than in the delayed neuronal death (+) subgroup (P 〈 0.05). The delayed neuronal death rates were 42% (5/12), 33% (4/12) and 33% (4/12) at 3, 7 and 30 days, respectively, followingischemia/reperfusion. No significant differences were detected at various time points (χ2 = 0.341, P 〉 0.05).
CONCLUSION: The activation of astrocytes was poor in the hippocampal CA1 region during the early stages of ischemia, which is an important reason for delayed neuronal death. Glial scar formation aggravated delayed neuronal death during the advanced ischemic stage.  相似文献   

3.
急性局灶性脑缺血后细胞凋亡的动态变化   总被引:4,自引:0,他引:4  
目的:细胞凋亡是细胞死亡的一种形式,对缺血后梗塞灶的发生,发展一定的作用。故测定缺血脑区出现DNA片断化的细胞数量变化及在缺血脑区的分布状况,以了解脑缺血的病理生理及寻求防治措施。方法:利用线栓法制成大脑中动脉缺血动物模型,测定脑组织SOD,LPO含量变化及凋亡细胞的密度变化;  相似文献   

4.
目的 探讨腺苷预处理对脑缺血再灌注损伤脑内星形胶质细胞的影响.方法 制作大鼠脑缺血再灌注损伤模型.60只SD大鼠随机分为3组:假手术组(F组)、缺血再灌注组(IR组)、腺苷预处理组(AP组),再按缺血再灌注后不同时间把各组随机分成4个亚组,每组5只大鼠.应用Zeal Longa 5级评分法进行神经功能评分,并通过免疫组织化学法检测脑组织内胶质纤维酸性蛋白(glial fibrillary acid protein,GFAP)的表达.结果 (1)神经功能评分AP组各亚组均小于IR组各亚组(P均<0.05),但大于F组各亚组(P均<0.05);(2)F组GFAP阳性表达均较弱,IR组和AP组在脑缺血再灌注后2h开始出现GFAP阳性表达的细胞数量增多,AP组在6h、24h AP组GFAP阳性表达比IR组增强(P均<0.05),在72h时AP组GFAP阳性表达较IR组减少(P<0.05).结论 腺苷预处理能在大鼠局灶性脑缺血再灌注损伤早期阶段促进GFAP的表达,72h后抑制GFAP的过度表达.  相似文献   

5.
目的 探讨小鼠大脑中动脉缺血再灌注后脑梗死体积、星形胶质细胞(AS)病理形态及其蛋白表达的变化.方法 选取69只成年健康雄性KM小鼠,将小鼠随机分为脑缺血2h再灌注1、4、10、24、48、72 h模型组(n=9)以及假手术组(n=6,仅结扎右侧颈总动脉)和正常组(n=9).采用线栓法制备小鼠大脑中动脉局灶性脑缺血再灌注模型.应用2,3,5-氯化三苯基四氮唑(TTC)染色法观察脑梗死的部位与体积;免疫组化法观察脑缺血中心区与周边区不同再灌注时间点胶质纤维酸性蛋白(GFAP)的表达.结果 缺血再灌注1h开始出现脑梗死灶,且再灌注24 h时梗死体积最大,之后逐渐缩小(F=745.534,P=0.000).小鼠脑缺血再灌注后缺血中心区AS肿胀、肥大进而较快发生凋亡,其GFAP表达水平低于周边区(P<0.05);而缺血周边区GFAP阳性表达的AS出现反应性活化,进而发生形态学改变;缺血周边区及对侧相应脑组织区域GFAP的表达量均随再灌注时间的延长呈现增加趋势(P<0.05).结论 AS反应性活化可能是一种缺血后的全脑保护性防御机制,尤其是在缺血周边区,其活化程度影响脑组织的存活与修复,提示在脑缺血再灌注过程中AS反应性活化可能在脑损伤后可塑性形态及功能改变中发挥重要作用.  相似文献   

6.
目的 观察病变侧缺血至再灌期亚低温 (32~ 33℃ )对局灶脑缺血再灌注后梗死体积、星形胶质细胞胶质纤维酸性蛋白 (GFAP)表达的影响。方法 采用改良线栓法建立大鼠大脑中动脉缺血再灌注模型 ,缺血 30min后应用反馈控温半导体制冷块对大鼠病变侧给予亚低温治疗 ,并持续至再灌期。采用免疫组织化学方法检测GFAP表达 ,TTC染色测梗死体积。结果 同常温组相比梗死体积明显减少 (P <0 .0 5 ) ,缺血常温组GFAP阳性细胞数量增多 ,突体粗大 ,胞体肿胀 ;亚低温组GFAP表达数量明显减少。结论 病变侧亚低温能明显抑制脑缺血后星形胶质细胞反应性增生和肥大。缺血至再灌期亚低温明显减轻脑组织损伤。  相似文献   

7.
目的探讨脑缺血损伤后星形胶质细胞和神经元在时间和空间上的动态变化。方法健康雄性Wistar大鼠48只.随机分为假手术组、缺血0.5~24h组,采用大脑中动脉线栓模型.结合组织学和SP免疫组化方法,动态观察缺血不同时间神经元和星形胶质细胞分布、形态.通过组织化学染色,观察星形胶质细胞的周长和积分光度值的改变。结果星形胶质细胞对缺血极为敏感;缺血灶周边区和对侧半球反应性星形胶质细胞增生明显;反应性星形胶质细胞分布区神经元损伤较轻。结论反应性星形胶质细胞参与脑保护及促进损伤神经元修复。  相似文献   

8.
目的观察大鼠反复前脑缺血再灌注后不同脑白质区胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)表达的变化,探讨其规律,为对脑缺血后星形胶质细胞的进一步研究提供实验依据。方法反复夹闭大鼠双侧颈总动脉制备前脑缺血再灌注模型,免疫组化法检测脑缺血再灌注后1周、2周、4周胼胝体、内囊和脑室周围GFAP的表达。结果缺血再灌注后,不同部位各时间点GFAP的表达均高于假手术组水平;在胼胝体、内囊GFAP的表达在1周时增加,2周时持续上升,4周时更明显;而脑室周围则在1周时上升,2周时达高峰,4周时回落但仍高于1周时的水平。结论反复前脑缺血后白质区GFAP表达明显升高,但不同脑区变化的规律和幅度略有差异,说明不同脑区对缺血的敏感性不同,星形胶质细胞的反应性略有差异。  相似文献   

9.
猫持续性局灶脑缺血模型的改进   总被引:5,自引:0,他引:5  
目的对猫持续性脑缺血模型加以改进,使其制作简便、模型标准、稳定。方法猫麻醉后,经眶后暴露大脑中动脉,在其发出外侧纹状体动脉的外侧部电凝闭塞,以医用耳脑胶及明胶海绵粘合硬膜和骨窗,术后行HE染色,计算梗死体积百分比。结果手术后在猫大脑中动脉(MCA)皮层分布区均产生梗死灶,梗死体积为19.3±0.70,动物无死亡及并发症的发生。结论该模型制作方便,重复性及稳定性好,适合于缺血性脑血管疾病的研究。  相似文献   

10.
目的 为研究康复训练对大鼠脑梗死灶周围胶质纤维酸性蛋白 (GFAP)表达的影响。方法  4 0只Wistar大鼠采用 L onga颈外动脉线栓法制备大鼠大脑中动脉闭塞模型 ,随机分为康复组、造模对照组 ,康复组每天进行 1h滚筒、平衡木、转棒及网屏训练 ;造模对照组置于普通笼中饲养 ,不予以康复训练。每组随机分为 3d、7d、14 d、2 1d 4个时间点 ,每个时间点各 5只大鼠 ,分别于相应天数取脑 ,采用免疫组织化学方法观察每个时间点脑梗死灶周围 GFAP的表达。结果 康复组神经功能评分较造模对照组为好 (P<0 .0 5 ) ;GFAP阳性细胞于脑缺血 3d后即已出现 ,7d、14 d、2 1d大量表达 ,14 d为高峰 ,且康复组较造模对照组明显增多 (P<0 .0 5 )。随时间延长免疫组化染色变深。结论 康复训练能促进中枢神经系统表达较高水平的 GFAP。  相似文献   

11.
背景:突触是神经系统可塑性最强的部位,脑缺血可导致突触病理性和生理性的可塑性变化。针刺对大脑的功能重组和代偿起着重要作用,针刺对突触重建的促进作用是目前的研究热点。目的:观察电针对局灶性脑缺血大鼠缺血灶周围区皮层突触超微结构的影响,进一步揭示电针治疗脑缺血疾病的可能作用机制。设计、时间和地点:随机对照动物实验,于2008-01/2009-01在广州中医药大学动物实验中心和电镜室完成。材料:雄性Wistar大鼠90只,随机分为3组:假手术组、模型组和电针组,各组又分为术后l h,l,3,7及21 d组5个时间段亚组,6只/亚组。G-6805型电针仪为上海华谊仪器厂生产。JEM-1200EX透射电子显微镜为日本JEOL公司生产。方法:模型组和电针组采用热凝闭大脑中动脉建立局灶性脑缺血模型,假手术组只暴露大鼠大脑中动脉,不予凝闭。电针组术后立即给予电针治疗,取百会(GV20)、大椎(GV14)穴,选用疏密波(疏波: 4 Hz; 密波:20 Hz),电压2.0~3.0 V,电流强度1~3 mA,留针30 min,1次/d。其他2组不予针刺处理。主要观察指标:透射电镜观察各组大鼠脑缺血灶周围区皮层突触的超微结构及结构参数(突触后致密物厚度、突触间隙宽度及突触界面曲率)的变化。结果:电镜下可见缺血灶周围区皮层突触结构发生破坏,突触数目明显减少,电针组与模型组比较有显著改善。缺血性脑损伤后PSD厚度、突触间隙宽度及突触界面曲率都明显减小,而电针组的PSD厚度于各时段均明显高于同时段模型组(P﹤0.05,P﹤0.01),电针组的突触间隙宽度在1d、3d及7d时均明显高于同时段模型组(P﹤0.05),电针组的突触界面曲率在1d、3d、7d及21d时均明显高于同时段模型组(P﹤0.05,P﹤0.01)。结论:电针从脑缺血早期开始就能明显抑制缺血损伤所致的突触退化、崩解,促进突触结构的修复,提高突触界面结构参数,电针治疗脑缺血的机理可能与其促进突触重建有关。  相似文献   

12.
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.  相似文献   

13.
目的观察亚低温对大鼠脑缺血再灌注损伤后热休克蛋白70(HSP70)及胶质纤维酸性蛋白(GFAP)表达的影响。方法将雄性Wistar大鼠30只分为假手术组、常温组和亚低温组。制作右侧大脑中动脉阻塞(MCAO)模型,观察缺血2h再灌注48h后各组大鼠脑组织学改变和HSP70及GFAP的表达。结果常温组大鼠脑皮质下神经元严重坏死,亚低温组皮质下神经元坏死严重程度明显较常温组轻,假手术组未见神经元坏死。常温组大鼠脑组织GFAP和HSP70阳性细胞较多,假手术组、亚低温组GFAP和HSP70阳性细胞少于常温组,假手术组偶见HSP70阳性细胞;图像分析显示,常温组大鼠脑组织GFAP、HSP70表达的平均光密度较假手术组和亚低温组明显增高(均P<0.01)。结论亚低温能减轻大鼠脑缺血再灌注损伤,降低脑组织HSP70及GFAP蛋白的表达。  相似文献   

14.
The astrocyte is a critical regulator of neuronal survival after ischemic brain injury. Electroacupuncture may be an effective therapy for cerebral ischemia, as electroacupuncture frequency can affect the structural integrity of astrocytes. In this study, a rat model of middle cerebral artery occlusion established using the modified thread embolism method was treated with electroacupuncture of the bilateral Quchi (LI11) and Zusanli (ST36) at 15, 30, and 100 Hz frequencies. Behavioral testing, immunohistochemistry and electron microscopy were used to explore the effect of these electroacupuncture frequencies used on maintaining the structural integrity of ischemic brain tissue. Compared with the model and 100 Hz electroacupuncture groups, the 15 and 30 Hz electroacupuncture groups displayed decreased neurological deficit scores, as evaluated by the Longa method, significantly increased glial fibrillary acidic protein expression, and alleviated ultrastructural damage of astrocytes at the edge of the infarct. Our experimental findings indicate that 15 and 30 Hz electroacupuncture intervention can favorably maintain the structural integrity of astrocytes and play a protective role in cerebral ischemic injury. Astrocyte structural integrity may be the mechanism underlying acupuncture production of ischemic tolerance.  相似文献   

15.
观察脑出血急性期大鼠延髓内脏带 ( MVZ)内星形胶质细胞的反应。以尾壳核局部注射胶原酶制作脑出血模型 ,用抗星形胶质细胞特异性标识物胶质原纤维酸性蛋白 ( GFAP)的免疫细胞化学方法 ,研究脑出血后 MVZ内星形胶质细胞的变化。发现脑出血后 4h GFAP阳性细胞数量增多、胞体增大、突起伸长 ,在MVZ形成明显弧形带状分布 ,尤以 MVZ背内侧区、中间带及腹外侧区明显。提示 MVZ内星形胶质细胞可能参与了脑出血后的病理生理过程。  相似文献   

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BACKGROUND: The onset of focal cerebral ischemia activates extracellular signal-regulated kinases 1 and 2, regulates cell cycle, promotes cell proliferation and differentiation, and affects the normal stage and function of brain cells.OBJECTIVE: To observe the effects of electroacupuncture at the Ren channel on extracellular signal-regulated kinases 1/2 expression in the lateral cerebral ventricle wall of rats with focal cerebral ischemia. The effects were analyzed at different time points after intervention.DESIGN: Randomized controlled study.SETTING: Department of Anatomy, Sun Yat-Sen University. MATERIALS: A total of 60 healthy adult male Wistar rats weighing (250±10) g were provided by the Experimental Animal Center, Medical College of Sun Yat-Sen University. The animal experiment was conducted with confirmed consent by the local ethics committee. The GB6805-Ⅱ electric acupuncture apparatus was provided by Shanghai Medical Equipment High-techno Company. METHODS: The experiment was performed at the Laboratory of Anatomy, Sun Yat-Sen University, from February to July 2007. All experimental animals were randomly divided into the following groups: normal group (n = 6), sham operation group (n = 18), model group (n = 18), and electroacupuncture group (n = 18). Middle cerebral artery occlusion (MCAO) was performed in the model group and electroacupuncture group. Zea Longa's grading standard was used to assess neurological impairment after reperfusion; animals whose grades were between 1 and 4 were included in this study. The normal control group was not exposed to MCAO. In sham operation animals, the right common carotid artery (CCA) was isolated, and the external carotid artery (ECA) was damaged, but no embolism was induced. The electroacupuncture group was given acupuncture on the second day after surgery. The acupoint locations were chosen according to Experimental Acupuncture (People's Publishing House; 1997; First Edition). The Chengjiang, Qihai, and Guanyuan acupoints were labeled and connected to a G6805 electroacupuncture apparatus with sparse-dense waves (sparse waves were 30 Hz, dense waves were 100 Hz), with a frequency of 6-15 V. The duration was 20 minutes. Two days after surgery, the model and sham operation groups were placed with their backs on the operating table, but they received no acupuncture. However, the normal group received acupuncture. The experimental animals under anesthesia were sacrificed on days 7, 14, and 28 post-surgery. Western blot analysis was used to measure expression of extracellular signal-regulated kinases 1/2 in the inferior region of the lateral cerebral ventricle wall. Expression was measured in the normal group at time points corresponding to the sham operation group. MAIN OUTCOME MEASURES: Expression of extracellular signal-regulated kinases 1/2 in the inferior region of the lateral cerebral ventricle wall at different time points after intervention.RESULTS: All 60 rats were included in the final analysis, without any loss. Seven days after MCAO, there was no significant difference in extracellular signal-regulated kinases 1/2 expression in the electroacupuncture group compared to the model group (P > 0.05). However, extracellular signal-regulated kinases 1/2 expression significantly increased in the model group at 14 and 28 days after treatment (P < 0.05).CONCLUSION: Electroacupuncture at the Ren channel can enhance extracellular signal-regulated kinases1/2 expression in the inferior region of the lateral cerebral ventricle wall of rats with focal cerebral ischemia. However, this effect is not apparent until 14 days after electroacupuncture intervention.  相似文献   

18.
目的探讨急性脑梗死(ACI)患者血清胶质纤维酸性蛋白(GFAP)含量的动态变化及其临床意义。方法检测47例ACI患者发病<48h、第3d和第5d的血清GFAP含量,按牛津郡社区卒中项目(OCSP)和CT分型对患者的血清GFAP含量进行比较,并分析其与病情、预后的关系。结果血清GFAP含量在OCSP各亚型患者中,完全前循环梗死(TACI)组发病<48h时明显高于对照组和其他3组,第3d、第5d继续上升;部分前循环梗死(PACI)组在发病第3d达高峰,第5d下降至接近正常水平;而后循环梗死(POCI)组和腔隙性梗死(LACI)组与对照组相比差异无显著性(均P>0.05)。血清GFAP含量在不同面积ACI患者中,发病<48h时各组血清GFAP含量差异无显著性(P>0.05);大面积梗死组第3d、第5d血清GFAP含量明显高于其他4组(均P<0.05),其他4组血清GFAP含量差异均无显著性(均P>0.05)。血清GFAP含量与梗死灶部位无明显相关性,仅第3d的皮质 皮质下组GFAP含量显著高于同时间的其他3组。血清GFAP含量与国立卫生研究所卒中量表(NIHSS)评分呈显著正相关(r=0.410,P<0.01);在病情好转组、未变组及恶化组差异无显著性(均P>0.05),不是预后的独立危险因素。结论ACI后血清GFAP含量的变化可反映病灶大小和病情的严重程度,为指导临床的诊治提供依据。  相似文献   

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