贝伐单抗对放射性脑损伤大鼠的治疗作用

目的通过建立大鼠放射性脑损伤模型,研究贝伐单抗对放射性脑损伤的治疗作用及其机制。方法将20只SD大鼠随机分成实验组(n=10)和对照组(n=10),均采用大剂量伽玛射线照射,在1个月内建立放射性脑损伤模型。建模后,实验组大鼠采用贝伐单抗干预,对照组采用生理盐水干预。通过对大鼠体质量、行为学评分、MRI检查和脑组织免疫组化检测,进而评估贝伐单抗对放射性脑损伤大鼠的治疗作用。结果实验组体质量较对照组有升高,行为学评分增加(P0.05)。MRI检查显示:对照组可见大鼠全脑弥漫性肿胀,实验组可见脑室系统增大,水肿带消退。对照组大鼠脑组织可见血管内皮生长因子(VEGF)、血管内皮生长因子受体2(VEGFR2)及CD34过量表达,实验组VEGF、CD34无表达,VEGFR2有少量表达。结论大剂量伽玛射线照射可在短期内迅速建立弥漫放射性脑损伤模型,证实贝伐单抗可通过拮抗VEGF的生物学效应来治疗放射性脑损伤。     

大鼠弥漫性脑损伤阻滞ERK通路下调脑组织MMP-9 mRNA的表达

目的探讨大鼠弥漫性脑损伤后磷酸化ERK1／2和基质金属蛋白酶-9（MMP-9）的变化规律及相互作用关系，从而进一步理解ERK1／2在弥漫件脑损伤中的作用及其对脑的保护机制。方法按Mamarou方法制作大鼠重型弥漫性脑损伤模型，尾静脉沣射ERK通路阻滞剂U0126；Western blot法检测磷酸化ERK1／2；RT—PCR检测MMP-9 mRNA；干湿重法测脑组织含水量。结果弥漫性脑损伤后磷酸化ERK1／2表达迅速增高，持续高水平表达至72h。MMP-9 mRNA在损伤后3h开始上升，24h达高峰，可维持较高水平直至7d。注射U0126后，磷酸化ERK1／2表达明显下降（P〈0．01），MMP-9 mRNA表达亦明显下降（P〈0．05），脑组织含水量减少（P〈0．05）。结论大鼠重型弥漫性脑损伤后ERK1／2被过度激活。MMP-9 mRNA表达增高，通过阻滞ERK通路可以下调MMP-9 mRNA的表达．保护受损脑组织。     

颅脑损伤大鼠肠道防御素-5 mRNA表达

目的探讨弥漫性脑损伤后肠道防御素-5（RD-5）变化。方法大鼠32只，分为对照组8只；弥漫性脑损伤组24只，再分成3个亚组（24h，3d，7d）。观察回肠RD-5mRNA的表达，回肠肠粘膜的病理改变以及血液内毒素的变化。结果弥漫性脑损伤后24hRD-5 mRNA的表达显著升高（P〈0．01），伤后3d降低至正常水平以下；回肠肠粘膜损伤，血液内毒素伤后24h最高（P〈0．01），3至7d仍显著高于正常水平（P〈0．01）。结论弥漫性脑损伤早期RD-5 mRNA的表达增强可能是机体的一种保护性反应。     

大鼠弥漫性脑损伤后脑组织中MMP-9的表达与神经元损伤

目的探讨大鼠弥漫性脑损伤后基质金属蛋白酶-9（MMP-9）在脑组织中的表达与神经元损伤的关系。方法荧光实时定量RT—PCR和免疫组化分别测定大鼠弥漫性脑损伤后MMP-9 mRN和MMP-9在脑组织中的表达。光镜和电镜观察神经元的变化。结果外伤后3h大鼠脑组织中MMP-9的阳性表达率开始升高，72h达高峰（50．35％±7．27％），伤后6h～7d各组均明显高于假手术对照组（P〈0．05）；随后开始下降，14d降至对照组水平。MMP-9 mRNA于伤后1h表达水平开始升高，72h达最高（20．56±1．29），伤后12h～7d时明显高于假手术对照组（P〈0．01）；随后开始下降，14d降至对照组水平。光镜及电镜下均可见弥漫性神经元变性和坏死，伤后72h损伤表现最为严重。结论本研究提示大鼠弥漫性脑损伤后MMP-9表达升高并可能参与了外伤后炎症反应和神经元损伤的病理过程。     

NBQX对大鼠全脑照射后早期脑MyT1阳性细胞的保护作用

目的 探讨大鼠全脑照射后早期大脑皮质少突胶质前体细胞的放射性反应及MK-801和NBQX对其保护作用。方法 以10Gy的剂量时SD大鼠单次全脑照射后即刻分别向其大脑皮质定向注射5mmol／L的MK-801和50mmol／L的NBQX各1μl，然后用免疫荧光组织化学法分别检测各时间点大脑皮质MyT1阳性细胞数量。结果 和假照射组相比，照射组大鼠照射后7d和14d时大脑皮质MyT1阳性细胞数显增加(P＜0．01)；MK-801组大鼠照射后各时间点大脑皮质MyT1阳性细胞数与照射组大鼠无明显差异(P＞0．05)；而NBQX组大鼠照射后1d、7d和14d时的MyT1阳性细胞数明显多于照射组大鼠(P＜0．05)。结论 大鼠全脑照射后早期大脑少突胶质前体细胞反应性增多，并有时程性变化；NBQX对早期的少突胶质前体细胞放射性损伤可能有一定的保护作用。     

冷诱导RNA结合蛋白mRNA在低体温大鼠脑内的表达

目的 观察大鼠脑内不同脑区，不同低温条件下冷诱导RNA结合蛋白（CIRP）mRNA的表达情况。方法 利用麻醉配合体表降温，制作亚低温大鼠模型。将大鼠分为对照组、低温30min、低温1h、低温2h、低温4h组，每组5只。保持低温状态恒定后，在相应时间点处死大鼠．取下丘脑、海马、皮层脑组织。以PT—PCR半定量检测各脑区在不同低温时间点时CIRP mRNA的表达。结果 正常体温下，海马CIRP mRNA的表达相对最强，皮层居中，下丘脑最弱。低体温1h后，CIRP mRNA的表达在下丘脑首先增高；低体温2h后．海马和皮层的CIRP mRNA的表达才开始增高，此时下丘脑CIRP mRNA的表达未见变化：低体温4h后，海马CIRP mRNA的表达较2h时仍有所增高，而下丘脑和皮层则保持不变。结论 不同脑区CIRP mRNA的表达在低温下均明显增高，各脑区表达不一，提示这种CIRP的高表达可能参与了低温下机体对脑损伤的保护作用。     

大鼠半脑照射后海马细胞外液谷氨酸含量的变化

目的 :探讨早期放射性脑损伤的机制。方法 :用在体微透析结合高效液相色谱法 ,对大鼠半脑照射后测定海马细胞外液谷氨酸含量。结果 :发现半脑照射后半个月照射侧谷氨酸含量上升 ,并与剂量相关 ,15Gy组和 3 0Gy组与未照射侧相比有显著差异。照射后 3个月恢复正常。结论 :认为谷氨酸神经递质的变化可能是放射性脑损伤综合作用的结果。     

胶质瘤伽玛刀治疗后脑水肿与C-myc蛋白表达的关系

目的　观察胶质瘤Wistar荷瘤大鼠γ刀照射治疗后 ,出现放射性脑水肿的比例、程度及分布与γ刀照射前后C -myc基因蛋白水平表达变化的相关关系。 方法　建立颅内种植胶质瘤C6细胞的Wistar荷瘤大鼠模型。 3周后 ,实验组给予γ刀治疗。免疫组织化学SP染色及计算机图像分析检测γ刀照射前后大鼠C -myc蛋白表达水平及γ刀照射后大鼠脑水肿情况。结果　胶质瘤荷瘤大鼠C -myc基因蛋白水平阳性表达平均百分率 ,未经照射的对照组为 77.62± 12 .11% ;实验组中 ,明显脑水肿大鼠 (n=2 2 )平均为 5 4 .81± 2 0 .15 % ,无水肿大鼠 (n=18)平均为 2 7.15± 10 .42 %。明显脑水肿组分别与对照组及无水肿组比较 ,差异显著 (P <0 .0 1) ,且水肿组织聚集于C -myc蛋白阳性表达细胞周围。 结论　γ刀照射后 ,放射性脑水肿发生的机理与胶质瘤细胞C -mcy基因蛋白水平表达程度有关     

内源性VEGF表达对缺血性脑损伤后成年大鼠海马齿状回神经发生影响的实验研究

2-EGFP-U6-shRNA(VEGF)立体定向给药大鼠缺血性脑损伤后24h和6d时缺血性脑损伤诱导的大鼠海马VEGF mRNA表达水平反应性升高可被抑制(P<0.05),海马内源性VEGF mRNA表达水平的降低可显著下调缺血性脑损伤后齿状回神经前体细胞增殖水平.结论 内源性VEGF基因表达可能是缺血性脑损伤后齿状回神经前体细胞增殖的一个重要启动信号.VEGF是耦联缺血性脑损伤和齿状回神经发生的关键分子.     

Caspase-1抑制剂对大鼠外伤性脑损伤后脑水肿、神经细胞凋亡和IL-18表达的影响

目的研究天冬氮酸特异性半胱氨酸蛋白酶-1（Caspase-1）抑制剂对大鼠脑损伤后脑水肿、细胞凋亡和IL-18表达的影响。方法雄性sD大鼠170只，随机分为正常组、损伤组、治疗组。用改良的Feeney方法制备损伤模型。治疗组分别于损伤前后30min经右侧脑室注射Caspase-1抑制剂——z-YVAD—fmk。损伤组与治疗组分别于脑损伤后6、24、72、168h点断头取脑。检测脑组织含水量、Caspase-1与IL-18表达及细胞凋亡。结果伤后6、24、72及168h，损伤组脑损伤周围皮质组织含水量、Caspase-1与IL—18的表达及细胞凋亡较正常组明显增多（P〈0．05），但伤后6、24及72h，治疗组脑损伤周围皮质组织含水量、Caspase—1与IL-18的表达及细胞凋亡虽明显高于正常组，但与损伤组相比则明显降低（P〈0．05）。结论Caspase-1及其激活的细胞因子IL—18在创伤性脑损伤后脑水肿和细胞凋亡中起重要作用，Caspase-1抑制剂对脑创伤后神经细胞损害具有保护作用。     

Expression of P-selectin and intercellular adhesion molecule-1 in human brain after focal infarction or cardiac arrest

Data from experimental studies indicate that acute inflammation contributes to ischaemic brain damage. Tethering of neutrophils to brain endothelium is mediated by selectins, and subsequent adhesion and migration by endothelial intercellular adhesion molecule-1 (ICAM-1) and neutrophil CD18. In experimental studies of ischaemia-reperfusion injury, brain damage has been ameliorated by administration of antibodies to these adhesion molecules. We studied the expression of P-selectin and ICAM-1 in sections of brain from patients who had experienced cardiac arrest or focal brain infarction, and who died 3.5 h to 9 days later. Endothelial immunopositivity for both adhesion molecules was maximal at about 2-3 days then declined. Between 1 day and 3 days, P-selectin was also detected on platelets in blood vessels within infarcted tissue. Within infarcts, but not sections of brain from cardiac arrest patients, P-selectin and ICAM-1 were again detectable at 1 week, when hyperplastic endothelial cells were labelled in capillaries in and immediately adjacent to the infarcted tissue. The finding that P-selectin and ICAM-1 are upregulated within focally infarcted brain tissue supports the concept that blocking neutrophil adhesion may be of benefit in treating atherothrombotic strokes in man.     

Decreased expression of CD200 and CD200 receptor in Alzheimer's disease: a potential mechanism leading to chronic inflammation

Inflammatory activation of microglia in response to neurodegenerative changes in diseases such as Alzheimer's disease (AD) and Parkinson's disease has been extensively described. These observations have suggested that inflammation could be contributing to disease progression. In this paper, the potential role of CD200 and CD200 receptor (CD200R), whose known functions are to activate anti-inflammatory pathways and induce immune tolerance through binding of CD200 to CD200 receptor (CD200R), was studied in AD. Quantitative studies showed a significant decrease in CD200 protein and mRNA in AD hippocampus and inferior temporal gyrus, but not cerebellum. Immunohistochemistry of brain tissue sections of hippocampus, superior frontal gyrus, inferior temporal gyrus and cerebellum from AD and non-demented cases demonstrated a predominant, though heterogeneous, neuronal localization for CD200. Decreased neuronal expression was apparent in brain regions affected by AD pathology. There was also a significant decrease in CD200R mRNA expression in AD hippocampus and inferior temporal gyrus, but not cerebellum. Low expression of CD200R by microglia was confirmed at the mRNA and protein level using cultured human microglia compared to blood-derived macrophages. Treatment of microglia and macrophages with interleukin-4 and interleukin-13 significantly increased expression of CD200R. Expression of these cytokines was not generally detectable in brain. These data indicate that the anti-inflammatory CD200/CD200R system may be deficient in AD brains. Mechanisms aimed at increasing levels of CD200 and CD200R could have therapeutic potential for controlling inflammation in human neurodegenerative diseases.     

小脑变性相关蛋白2 mRNA在癫痫患者脑组织中的表达及其临床意义

目的了解小脑变性相关蛋白2(CDR2)mRNA在癫痫(EP)患者脑组织中的表达及其临床意义。方法将48例EP患者分成耐药(40例)和非耐药(8例)两组,用基因芯片扫描和FQ-PCR分别检测脑组织中CDR2基因表达,并与对照组比较。结果基因芯片扫描提示CDR2 mRNA在EP患者与对照组中的表达差异有统计学意义(P<0.05)。FQ-PCR结果与基因芯片扫描一致。结论EP患者脑组织中有CDR2 mRNA表达,由于其表达产物可通过血-脑脊液屏障进入脑脊液(CSF)和血液中,因而检测CDR2 mRNA在血或CSF中的表达产物有可能成为诊断EP的一个重要指标。     

CD47 gene knockout protects against transient focal cerebral ischemia in mice

CD47 is a cell surface glycoprotein that helps mediate neutrophil transmigration across blood vessels. The present study was performed to determine whether absence of the CD47 gene decreases focal ischemic brain damage. Mice were subjected to 90 min middle cerebral artery occlusion. CD47 knockout mice were compared against matching wildtype mice. CD47 expression was checked by Western blotting. Infarct volume and ischemic brain swelling were quantified with cresyl violet-stained brain sections at 24 and 72 h after ischemia. The tight junction protein claudin-5 was detected by imunohistochemistry. Two surrogate markers of neuroinflammation, brain levels of matrix metalloproteinase-9 (MMP-9) and infiltration of neutrophils, were assessed by immunohistochemistry. Western blots confirmed that CD47 was absent in knockout brains. Ischemia did not appear to upregulate total brain levels of CD47 in WT mice. In CD47 knockout mice, infarct volumes were reduced at 24 and 72 h after ischemia, and hemispheric swelling was decreased at 72 h. Loss of claudin-5 was observed in ischemic WT brain. This effect was ameliorated in CD47 knockout brains. Extravasation of neutrophils into the brain parenchyma was significantly reduced in CD47 knockout mice compared to wildtype mice. MMP-9 appeared to be upregulated in microvessels within ischemic brain. MMP-9 levels were markedly lower in CD47 knockout brains compared to wildtype brains. We conclude that CD47 is broadly involved in neuroinflammation, and this integrin-associated-protein plays a role in promoting MMP-9 upregulaton, neutrophil extravasation, brain swelling and progression of acute ischemic brain injury.     

大鼠实验性脑出血后脑组织中c-fos基因的表达与局部脑血流的改变

目的 探讨大鼠实验性脑出血后脑组织中即刻早期基因c-fos的表达和局部脑血流的变化。方法 采用Nath改良法建立大鼠脑出血模型；免疫组化法及RT－PCR法测定其脑组织中fos蛋白和c-fos mRNA的表达；氢清除法测定其局部脑血流。结果 大鼠血肿周围区（基底节）在脑出血后1小时即出现fos蛋白的表达，至3小时达高峰；c-fos mRNA于出血后1小时达表达高峰,至3小时后仍有较高水平的表达；出血后1小时全脑的的血流量均下降，4小时恢复至对照组水平，并维持至出血后24小时，随着的24小时内再次出现脑血流下降。结论 大鼠脑出血后，血肿周围区和双侧皮质区的脑组织中存在着c-fos基因的快速而长久的诱导表达。局部脑血流的下降相对短暂，且脑血流的下降在时程上与c-fos基因的表达不相一致。     

人脑胶质瘤EPO与CD105表达及意义

目的 探讨人脑胶质瘤组织促红细胞生成素（EPO）和CD105的表达变化及其意义。方法 收集2002~2008年人脑胶质瘤标本152例,其中WHO分级Ⅰ级4例,Ⅱ级32例,Ⅲ级68 例,Ⅳ级48例;取同期颅脑损伤内减压术切除正常脑组织20例为对照,采用免疫组化染色分析EPO及CD105表达。收集2005~2008年人脑胶质瘤标本胶质瘤17例（WHO Ⅱ级6例,Ⅲ~Ⅳ级11例）,正常对照5例,采用实时荧光定量PCR检测EPO mRNA表达变化。术后随访截止2010年4月23日,应用Kaplan-Meier生存曲线分析高级别（Ⅲ~Ⅳ级）胶质瘤生存曲线。结果 胶质瘤EPO表达阳性率（60.5%,92/152）明显高于正常脑组织（10%,2/20;P<0.001）,胶质瘤EPO表达强度与病理分级呈正相关（rs=0.368,P<0.001）。EPO表达阳性组CD105阳性率明显高于阴性组（P<0.05）,EPO高表达组明显高于低表达组（P<0.05）。Ⅱ级胶质瘤组EPO mRNA表达水平明显高于正常组与Ⅲ~Ⅳ级胶质瘤组（P<0.05）。对于高级别（WHO Ⅲ~Ⅳ级）胶质瘤,EPO低表达组中位生存时间为12个月,高表达组为36个月;EPO低表达组累积生存率明显低于高表达组（P<0.05）。结论 人脑胶质瘤EPO蛋白的表达与病理级别及新生血管正相关;WHO Ⅱ级胶质瘤EPO mRNA在转录水平已上调;WHO Ⅲ~Ⅳ级组胶质瘤EPO表达高者生存期长。     

The leukocytes expressing DARPP-32 are reduced in patients with schizophrenia and bipolar disorder

Bipolar disorder (BPD) and schizophrenia (SCZ) are severe disorders representing an enormous social, familiar and individual burden, being SCZ the most disabling psychiatric disorder characterized by psychosis and cognitive impairment. It is well known that SCZ and BPD are associated with abnormalities in dopamine signaling pathway. Recent data in the literature have demonstrated altered expression levels of some proteins involved in the modulation of this pathway in both brain and peripheral tissues. It was shown that protein and mRNA levels of dopamine and cAMP regulated phosphoprotein (DARPP-32) were downregulated in dorsolateral prefrontal cortex (DLPFC) of patients with SCZ or BPD when compared to controls. Due to the difficulty to access brain tissue and the absence of objective laboratory tests for bio-markers, we measured DARPP-32 expression in blood cell sub-populations (CD4+ T lymphocytes, CD56+ NK cells, CD19+ B lymphocytes and CD14+ monocytes) taking advantage of the close relation of nervous and immune systems. Using flow cytometry as the analytical method, our results have shown that the DARPP-32 expression was diminished in CD4+ T lymphocytes, CD19+ B lymphocytes and CD14+ monocytes of BPD patients and was also decreased in CD4+ T lymphocytes and CD56+ NK cells of SCZ patients. These results showed that DARPP-32 expression in immune cells agrees with reports of reduced DARPP-32 protein in the DLPFC of BPD or SCZ patients. Our data suggest that DARPP-32 expression in PBMC could be used as a source of bio-markers to help in the treatment response of neuropsychiatry disorders as a window to the changes in the brain of those patients.     

RhoE在小鼠中枢神经系统通过NF-κB信号通路调节神经细胞凋亡

目的 探讨RhoE对神经细胞凋亡的作用及可能的相关机制。方法 用免疫组织化学染色、Q-PCR和免疫印迹等方法检测RhoE^-/-及RhoE^+/+小鼠脑组织内RhoE、Caspase3和P65蛋白表达水平。结果 RhoE^-/-小鼠脑组织内无RhoE蛋白表达; RhoE^-/-小鼠多部位脑组织cleaved-Caspase3表达水平较RhoE^+/+小鼠显著降低(P<0.05); 同时RhoE^-/-小鼠脑组织内P65蛋白表达水平较RhoE^+/+小鼠显著增高(P<0.05),但mRNA水平无明显改变(P>0.05)。结论 RhoE在小鼠中枢神经系统内可能通过NF-κB信号通路中P65蛋白表达水平来调控神经元的凋亡; RhoE可能是一个新的神经元凋亡调控位点。     

肿瘤转移抑制基因KiSS-1在乳腺癌脑转移患者的表达及临床意义

目的 转移抑制基因KiSS-l在多种肿瘤的浸润转移中起着重要的作用。但该基因与乳腺癌脑转移的关系仍很不清楚。本研究检测了乳腺癌原发灶和脑转移灶中KiSS-l基因的表达并探讨其临床意义。方法 选择2002年6月～2004年6月行乳腺癌脑转移病灶切除的患者12例，应用实时荧光定量PCR检测乳腺癌原发灶和转移灶中KiSS-l基因mRNA的表达，应用Westernblot检测KiSS-l蛋白的表达，并进一步应用免疫组化进行验证。结果 实时荧光定量PCR显示脑转移标本KiSS-l mRNA表达仅为原发灶的1／10，与原发灶比较有显著差异（P〈0．01），Western blot检测显示脑转移灶KiSS-l蛋白较原发灶明显减弱，免疫组化显示KiSS-l在原发灶中表达率明显低于原发灶。结论 KiSS-l基因在乳腺痛脑转移中具有转移抑制作用，可能在乳腺癌脑转移治疗中发挥作用。     

锌离子螯合剂DEDTC对MCAO大鼠的神经功能损害及AMPK/eNOS/NF-κB通路的调节作用

目的 探讨锌离子螯合剂DEDTC对大脑中动脉闭塞(MCAO)大鼠的神经功能损害及AMPK/eNOS/NF-κB通路的调节作用。方法 将雄性SD大鼠随机分为对照组、MCAO组、DEDTC组,后2组采用线栓法建立MCAO模型,DEDTC组在造模前30 min给予DEDTC干预,比较3组大鼠神经功能评分、血清生长细胞因子及炎症细胞因子水平、脑组织AMPK/eNOS/NF-κB通路的差异。结果 造模6、12、24 h后MCAO组大鼠的神经功能评分明显高于对照组,DEDTC组大鼠的神经功能评分明显低于MCAO组; 造模24 h后MCAO组大鼠的血清BDNF、VEGF、bFGF、TNF-α、IL-1β、IL-6、ICAM-1水平及脑组织中AMPK、eNOS、NF-κB的mRNA表达水平明显高于对照组; DEDTC组大鼠的血清BDNF、VEGF、bFGF水平以及脑组织中AMPK、eNOS的mRNA表达水平明显高于MCAO组,血清TNF-α、IL-1β、IL-6、ICAM-1水平及脑组织中NF-κB的mRNA表达水平明显低于MCAO组。结论 锌离子螯合剂DEDTC对MCAO大鼠神经功能损害具有抑制作用,且该作用可能与AMPK/eNOS/NF-κB通路介导的细胞因子分泌改变有关。