大鼠机械性脑损伤蛋白质表达谱的变化及意义

目的研究大鼠脑损伤后血清和脑干中蛋白质表达的特点。方法采用弱阳离子交换芯片（WCX-2）和金属亲和吸附芯片（IMAC-Cu）结合表面增强激光解析电离飞行时间质谱技术分析大鼠闭合性脑损伤后4h、8h、12h、24h、48h血清和脑干中蛋白质表达谱的改变。结果（1）在WCX-2芯片上,血清中共捕获436个蛋白质峰;脑干中共捕获495个蛋白质峰;血清和脑干都能捕获的蛋白质数是33个。在IMAC—Cu芯片上,血清中共捕获229个蛋白质峰;脑干中共捕获107个蛋白质峰;血清和脑干在IMAC—Cu芯片上都能捕获的蛋白质数是3个。（2）与手术对照组相比,血清在WCX-2芯片上损伤组共有53个蛋白质峰的相对强度有显著性差异（P〈0．05）;在IMAC-Cu芯片上损伤组共有16个蛋白质峰的相对强度有显著性差异（P〈0．05）。脑干在WCX-2芯片上损伤组共有25个蛋白质峰的相对强度有显著性差异（P〈0．05）;在IMAC．Cu芯片上损伤组共有2个蛋白质峰的相对强度有显著性差异（P〈0．05）。结论（1）脑损伤可引起血清和脑干蛋白质表达谱变化,提示血清中出现的部分蛋白质可能为脑损伤诱导的血细胞基因表达产物。（2）血清和脑干蛋白质表达谱存在差异。（3）在血清和脑干中检测到的差异蛋白质以及损伤后新出现的蛋白质峰,可能是脑损伤生物标志蛋白,对其进行进一步研究,有望深人了解脑损伤的分子机制,并用于脑损伤的诊断和治疗。     

大黄多糖对脑损伤后大鼠脑皮层热休克蛋白70表达的影响

目的：观察大黄多糖（Tanguticum Maxim polysaccharides,TMP)对大鼠脑损伤后不同时间热休克蛋白（heat shock protein,HSP)70表达的影响，探讨大黄多糖的脑保护作用机制，为其用于临床治疗脑损伤寻求依据。方法；复制大鼠脑挫裂伤模型，分别在伤后6，24，48h以免疫组织化学方法检测大脑皮层表达HSP70的神经细胞，以Western-blot蛋白印迹方法检测脑皮层HSP70表达变化。结果：大黄多糖治疗组6h HSP 70表达高于损伤组，24，48h均低于治疗组.结论：大黄多糖能促进HSP 70的表达，这可能是其脑保护作用的机制之一。     

缺氧缺血性脑损伤对新生大鼠发育期间额叶皮质突触质量的影响

目的：探讨缺氧缺血性脑损伤(hvpoxia ishemia brain damage，HIBD)对新生大鼠发育期间额叶皮质突触质量的影响：方法：取7日龄Wistai仔鼠80只，采用抽签法将每窝仔鼠随机分成实验组(E组)和假手术组(对照组C组)。实验组鼠在乙醚麻醉下行颈前正中切口，分离右侧颈总动脉，1号手术线结扎，保温2h后，放人含8％氧气的氮氧混合气体的容器内2h，制成HIBD模型。假手术组只分离该血管，不结扎也不低氧处理。从生后7d到2个月(成年)分8个时段对额叶皮质的发育，采用形态学和形态计量法的方法进行长期跟踪研究，结果：实验组6h～7d各组可见神经元、胶质细胞及毛细血管间隙变大，神经元胞质内出现空泡，线粒体肿胀、嵴模糊，RER肿胀脱颗粒，核皱缩不规则甚至溶解，神经元突起比假手术组稀疏。脑损伤后仔鼠额叶皮质神经元及神经纤维的密度显著降低(P&;lt;0．05，P&;lt;0．01)，突触的体视学参数，表面积密度和面数密度降低，突触后膜致密层变薄(P&;lt;0．05．P&;lt;0．01)。结论：缺氧缺血性脑损伤对新生大鼠发育期间额叶皮质中突触的质量，神经元及神经纤维密度的影响，是影响智力发育的重要因素。     

液相蛋白质芯片检测血清C反应蛋白方法的建立

目的 建立液相蛋白质芯片检测血清C反应蛋白（CRP）的方法,探讨该方法诊断冠心痛的临床应用价值。方法 用聚苯乙烯荧光微珠,将抗CRP单克隆抗体包被在微珠上,将另一针对CRP不同抗原表位的单克隆抗体生物素化;包被好的微珠加入到96孔板中,将待测血清分别加入各孔,用CRP标准品建立标准曲线,各孔中加入生物素化抗CRP单抗和PE荧光素标记的链霉亲合素。在Bio-Plex液相蛋白质芯片分析仪上检测296份血清样品。同一血清标本同时用免疫比浊法测定。比较两种方法对CRP检测的敏感性和特异性。结果 液相蛋白质芯片检测CRP方法对诊断冠心病的敏感性（78,3％）和特异性（87．1％）较免疫比浊法的敏感性（63．0％）和特异性（86．5％）高,χ^2=28,94,P〈0．001。结论 建立了一种检测血清CRP的新方法,对诊断冠心痛有临床应用价值。     

光学蛋白质芯片法定量检测C-反应蛋白的研究

目的本研究利用光学蛋白质芯片系统定量检测心肌梗死早期标志物C-反应蛋白（CRP）,探讨光学蛋白质芯片在医学检验诊断中的应用。方法实验采用椭偏光学生物显微成像系统、光学蛋白质芯片分析系统及软件、芯片装置、微流控芯片加样系统等,在4×6点芯片上装配生物探针,然后用人血清蛋白（HAS）封闭检测位点,利用抗原、抗体特异性结合的特性定量检测CRP（选用浓度为200、100、50、20、10、5μg/ml）,并绘制了检测CRP的标准曲线。采用SPSS统计学软件10.0分析处理实验数据。结果检测结果显示,CRP浓度与检测位点具有高度相关性,具有浓度-灰度的一致性,可以绘制出定量检测标准曲线,检测灵敏度为5μg/ml,符合临床检测CRP的灵敏度要求。各浓度（5、10、20、50、100、200μg/ml）的组内变异系数（CV%）分别为：2.47%、1.15%、2.06%、4.32%、1.39%、0.36%,皮尔逊相关系数为95.3%。结论光学蛋白质芯片可以定量检测CRP,灵敏度为5μg/ml,符合临床检测CRP的灵敏度和实用要求,具有临床检测CRP的潜在应用前景。此外,利用椭偏光显微成像技术的光学蛋白质芯片不需要标记试剂,被测样品不需要任何处理,样品需要量少,操作简单。     

大鼠液压脑损伤后fos蛋白表达与细胞凋亡

目的 探讨颅脑损伤后神经细胞凋亡分子的病理机制，为临床防治提供实验依据。方法 采用免疫组织化学技术和原位末端标记法(TUNEL)检测大鼠伤灶边缘皮层的fos蛋白及凋亡细胞。结果外伤组在伤后24h检测到高水平表达的fos蛋白及明显增多的凋亡神经细胞。结论 外伤后fos表达参与诱导了神经细胞凋亡过程。     

脂多糖与缺氧缺血对新生大鼠脑损伤的协同作用

目的：探讨亚临床感染是否与轻度缺氧缺血（HI）协同导致严重的脑损伤。方法：（1）用不同剂量脂多糖（LPS）腹腔注射7d龄Wistar大鼠，观察体温变化以确定引起亚临床感染的LPS剂量；（2）LPS 0.3mg/kg腹腔注射7d龄Wistar大鼠并联合不同时间HI，用微管相关蛋白－2（MAP－2）免疫组化染色测脑梗死面积；（3）用7d龄Wistar大鼠分别制成缺氧缺血损伤（HIBD）模型（HI 55min)、感染与轻度HI协同作用脑损伤模型（LPS＋HI 20min)和对照组，分别用MAP－2和细胞黏附因子－1（ICAM－1）免疫组化染色测脑梗死面积和脑HCAM－1阳性微血管计数。结果：（1）LPS 0.3mg／kg是引起亚临床感染的剂量；（2）LPS 0.3mg/kg HI 20min可造成严重脑损伤；（3）LPS＋HI 20min组与HI 55min组脑梗死面积和脑组织ICAM－1阳性微血管计数明显增加，与对照组相比有显著差异，（P＜0．05），LPS＋HI 20min组脑梗死面积与HI 55min组相比无显著差异（P＞0．05），ICAM－1阳性微血管计数LPS＋HI 20min组显著高于HI 55min组（P＜0．05）。结论：LPS与HI可协同导致严重脑损伤；ICAM－1在此过程中起重要作用；临床中对轻度窒息患儿应警惕感染／亚临床感染对HI脑损伤的协同作用。     

蛋白质芯片技术检测肿瘤标志物及临床应用

我科用蛋白质芯片对2 2 6例患者进行了肿瘤标志物检查,本文对检查结果进行回顾性分析。1　材料和方法1 1　标本来源　待测血清获自2 2 6例住院、门诊临床诊断疑似肿瘤患者,男性12 5例、女性10 1例,年龄(5 3±2 6 )岁。以手术所见和病理检验结果为诊断各类肿瘤金标准。1 2　仪器     

氯胺酮对大鼠蓝斑核和内侧前额叶皮质C-Fos蛋白表达的影响

目的:检测氯胺酮对大鼠蓝斑核(locus coeruleus,LC)和内侧前额叶皮质(medial prefrontalcortex,mPFC)c-Fos蛋白表达的影响。方法:24只Wistar大鼠随机分为4组(n=6):对照1组、2组(C1组、C2组),氯胺酮1组、2组(K1组、K2组),分别腹腔注射生理盐水,氯胺酮100 mg/kg。C1组和K1组用于检测大鼠用药120 min内的行为学变化和第120 min时的热痛阈值;C2组和K2组用于通过Western blotting方法检测用药第120 min时,LC和mPFC脑区Fos蛋白的表达水平。结果:K1组较C1组热痛阈值明显升高(P<0.05),运动亢进、刻板行为也有显著差异(P<0.01)。K2组较C2组LC和mPFC处Fos蛋白表达均明显增多(P<0.05和P<0.01)。结论:氯胺酮可引起热痛镇痛效应和行为异常,伴有LC和mPFC的Fos蛋白表达增加。     

Treatment of penetrating brain injury in a rat model using collagen scaffolds incorporating soluble Nogo receptor

Injuries and diseases of the central nervous system (CNS) have the potential to cause permanent loss of brain parenchyma, with severe neurological consequences. Cavitary defects in the brain may afford the possibility of treatment with biomaterials that fill the lesion site while delivering therapeutic agents. This study examined the treatment of penetrating brain injury (PBI) in a rat model with collagen biomaterials and a soluble Nogo receptor (sNgR) molecule. sNgR was aimed at neutralizing myelin proteins that hinder axon regeneration by inducing growth cone collapse. Scaffolds containing sNgR were implanted in the brains of adult rats 1 week after injury and analysed 4 weeks or 8 weeks later. Histological analysis revealed that the scaffolds filled the lesion sites, remained intact with open pores and were infiltrated with cells and extracellular matrix. Immunohistochemical staining demonstrated the composition of the cellular infiltrate to include macrophages, astrocytes and vascular endothelial cells. Isolated regions of the scaffold borders showed integration with surrounding viable brain tissue that included neurons and oligodendrocytes. While axon regeneration was not detected in the scaffolds, the cellular infiltration and vascularization of the lesion site demonstrated a modification of the injury environment with implications for regenerative strategies. Copyright © 2012 John Wiley & Sons, Ltd.     

急性颅脑损伤患者血清髓鞘碱性蛋白测定的意义

目的：探讨急性颅脑损伤时血清髓鞘碱性蛋白（ＭＢＰ）含量变化与颅脑损伤程度的关系。方法：采用酶联免疫吸附检测法（ＥＬＩＳＡ）测定１００例急性颅脑损伤患者入院时血清中ＭＢＰ含量，另测定５０例健康人血清ＭＢＰ含量为正常对照组。结果：急性颅脑损伤组血清ＭＢＰ含量（５．０２７±３．２３７）ｎｇ／Ｌ与正常对照组（１．０１７±０．６４２）ｎｇ／Ｌ比较有显著性差异（Ｐ＜０．０１）。不同病情组间血清ＭＢＰ含量〔轻型组：（２．５８１±１．０２２）ｎｇ／Ｌ，中型组（４．９９９±０．８３４）ｎｇ／Ｌ，重型组（９．９７５±３．５５３）ｎｇ／Ｌ〕比较均有显著性差异（Ｐ均＜０．０１）。结论：急性颅脑损伤时血清ＭＢＰ含量明显升高；脑实质损伤程度越重，其血清ＭＢＰ含量升高愈明显；及时测定脑外伤患者血清ＭＢＰ含量对判断脑损害程度及其预后有重要意义。     

牛脑提取液治疗大鼠脊髓损伤的实验研究

目的：探讨牛脑提取液对实验性大鼠脊髓损伤的作用饥制：方法：健康SD大鼠64只，随机分为硬膜内给牛脑提取液组、硬膜内给生理盐水组、腹腔内给牛脑提取液组、腹腔内给生理盐水组。采用改良的Allen氏装置制作模型：运用脊髓神经功能、体感诱发电位、辣根过氧化酶示踪、大体观察、组织学及超微结构变化作为观察指标进行实验观察。结果：实验组与对照组相比．其各项观测指标皆有明显改善；局部用药组观察指标明显优于全身用药组。结论：牛脑提取液无论局部应用或全身应用均能促进大鼠脊髓损伤后神经纤维的修复与再生，但以硬膜内给药为佳。     

局部用地塞米松对外伤性脑水肿和髓鞘碱性蛋白变化的影响

目的 :研究脑皮质局部应用地塞米松对兔脑外伤后脑水肿和脑组织损害的减轻程度。方法 :2 2只兔随机分为 A(对照组 )和 B(治疗组 ) 2组。应用骨窗成形硬脑膜外打击法制备脑挫裂伤动物模型。B组采用局部微量等距离扩渗法于脑皮质局部损伤灶内注射地塞米松 ;A组以同样方法注射等量生理盐水。以脑组织含水量和血清髓鞘碱性蛋白 (MBP)作为观察指标。结果 :A、B2组兔患侧大脑半球含水量分别为 (81.75± 0 .5 6 ) %和(79.45± 0 .5 2 ) % ,B组显著低于 A组 (P<0 .0 5 )。 A组血清 MBP为 (5 .98± 2 .0 8) μg/L,B组为 (3.15±1.0 9)μg/L ,A、B 2组 MBP均较正常对照值 (1.6 6± 0 .71)μg/L增高 ,但 A组显著高于 B组 (P<0 .0 5 )。结论 :脑挫裂伤后脑含水量和血清 MBP增高 ,治疗后两者均降低 ,说明局部应用地塞米松对脑损伤有明显治疗作用     

Serum S-100B protein monitoring in patients with severe traumatic brain injury

Objective S-100B protein is a promising marker of injury severity and outcome after head injury. We examined the relationship between serum S-100B concentrations and injury severity, clinical course, survival, and treatment efficacy after severe traumatic brain injury (TBI). Design and setting Prospective observational study in a neurosurgical intensive care unit. Patients and participants 102 adult patients with severe TBI, admitted between June 2001 and November 2003 (30 months). Interventions Serum S-100B levels were measured by immunoluminometric technique on admission and every 24 h thereafter for a maximum of 7 days. Measurements and results Initial S-100B levels were significantly related to pupillary status, computed tomography severity1, and 1-month survival. Cox's proportional hazard regression analysis showed that initial S-100B was an independent predictor of 1-month survival, in the presence of dilated pupils, and with increased age. Subjects with initial levels above 1 μg/l had a nearly threefold increased probability of death within 1 month. Serum S-100B alteration indicated neurological improvement or deterioration. Finally, surgical treatment reduced S-100B levels. Conclusions Serum S-100B protein reflects injury severity and improves prediction of outcome after severe TBI. S-100B may also have a role in assessing the efficacy of treatment after severe TBI.     

Diffuse and spatially variable white matter disruptions are associated with blast-related mild traumatic brain injury

Mild traumatic brain injury (mTBI) due to explosive blast is common among military service members and often associated with long term psychological and cognitive disruptions. Little is known about the neurological effects of blast-related mTBI and whether they differ from those of civilian, non-blast mTBI. Given that brain damage from blasts may be diffuse and heterogeneous, we tested the hypothesis that blast mTBI is associated with subtle white matter disruptions in the brain that are spatially inconsistent across individuals. We used diffusion tensor imaging to examine white matter integrity, as quantified by fractional anisotropy (FA), in a group of American military service members with (n = 25) or without (n = 33) blast-related mTBI who had been deployed as part of Operation Iraqi Freedom or Operation Enduring Freedom. History of civilian non-blast mTBI was equally common across groups, which enabled testing of both blast and non-blast mTBI effects on measures sensitive to (1) concentrated, spatially consistent (average FA within a region of interest [ROI]), (2) concentrated, spatially variable (number of ROIs with low average FA), and (3) diffuse (number of voxels with low FA) disruptions of white matter integrity. Blast mTBI was associated with a diffuse, global pattern of lower white matter integrity, and this pattern was not affected by previous civilian mTBI. Neither type of mTBI had an effect on the measures sensitive to more concentrated and spatially consistent white matter disruptions. Additionally, individuals with more than one blast mTBI tended to have a larger number of low FA voxels than individuals with a single blast injury. These results indicate that blast mTBI is associated with disrupted integrity of several white matter tracts, and that these disruptions are diluted by averaging across the large number of voxels within an ROI. The reported pattern of effects supports the conclusion that the neurological effects of blast mTBI are diffuse, widespread, and spatially variable.     

Tau protein as a serum marker of brain damage in mild traumatic brain injury: Preliminary results

The objective of this study was to investigate the diagnostic value of serum tau protein in determining the severity of traumatic brain injury in patients with mild traumatic brain injury (mTBI) and high-risk patients. Adult patients who presented to our emergency department (ED) with mTBI over 1 year were prospectively enrolled. Patients underwent cranial computed tomography (CT) and were subdivided into high and low-risk groups, according to the probability of resultant intracranial injury. Serum tau levels of 60 patients and 20 healthy volunteers, who served as a control group, were measured. The mean age of the 60 patients (45 males, 15 females) was 32.5 years (range, 15–66 y). Mean Glasgow Coma Scale (GCS) score was 14±0.6. CT scans demonstrated intracranial injury in 11 patients (18.3%) and depressed fracture in 4 patients (6.7%). Serum tau levels of patients (188±210 pg/mL), compared with those of controls (86±48 pg/mL), were relatively higher; however, differences were not statistically significant (P=.445). Also, serum tau levels of high-risk patients (307±246 pg/mL) were significantly higher than those of low-risk patients (77±61 pg/mL) (P=.001). A total of 48 patients (80%) were accessible for follow-up after 6 months. Postconcussive syndrome was observed in 8 patients, 5 of whom had serum tau protein levels that were higher than those of the other 3 patients. However, no statistically significant difference was observed (P > .05). Investigators of the present study noted that serum tau levels in patients with mTBI were increased. Therefore, it is believed that this biomarker may prove helpful in identifying high-risk patients with mTBI. However, additional studies are needed to establish the diagnostic value of serum tau in detecting traumatic brain injury in patients with mTBI.     

Participation in the workforce after a traumatic brain injury: a matter of control

Purpose: This study sought to explore individual experience in developing a mastery of daily activities and roles after a traumatic brain injury (TBI) with the objective of returning to work. Method: Eight 30–60-year-old men, employed at the time of injury, were each interviewed three times over a 6-month period. Ten to 21 months after the injuries, four participants had returned to work at least part time. Grounded theory was adapted for analyses. Results: A single core category emerged: a desire for control: focusing on high-priority issues. Still, 2 years after injury, the participants were uncertain about their abilities with respect to what was expected of them at work. They felt they would do better as time progressed. Conclusions: The participants’ uncertainty about their efficacy cast doubt on their beliefs in improving their skills, balancing daily activities and work. They wondered about the sustainability of their health and efficacy at work. Wanting to control their own improvement, the participants asked for counselling in strategies and techniques to help with their progress. This issue could be taken into account in follow-up rehabilitation programmes. Additionally, the workplace might be the ideal context in which to develop the structures and routines necessary to master life in general.

• Implications for Rehabilitation

• Two years after injury, the participants remained uncertain about their abilities with respect to what was expected of them at work. The participants felt they would do better as time progressed.

• The participants, wanting to control their own improvement, sought counselling to help sort out their priorities and found it could contribute to help with their progress in finding a suitable balance between daily activities and work.

• A consequence of our main finding, in a multidisciplinary context, is that counselling in structures and routines with respect to work-related tasks should be considered to be an integral part of any rehabilitation programme after TBI.

     

颅脑损伤患者血清C-反应蛋白和血糖水平的动态变化及其意义

目的研究颅脑损伤患者血清C-反应蛋白（CRP）、血糖（Glu）水平的动态变化及其意义。方法97例颅脑损伤患者按照病情计分法分为重型组（29例）、中型组（31例）、轻型组（37例）3组,35例健康体检者为健康对照组。用生化分析仪测定颅脑损伤患者伤后6h、1d、3d、14d血清中CRP、Glu水平。结果颅脑外伤患者伤后6h血清CRP、Glu含量均有不同程度升高,与健康对照组比较差异均有统计学意义（P〈0．01）。颅脑损伤患者CRP和Glu水平与患者入院时的GCS评分呈负相关,与其病情呈正相关,重型组患者血清cRP、Glu升高最为明显;中型和轻型组次之,3组之闻的差异有显著性;病情重和预后差的患者其CRP、Glu水平升高幅度大且持续时间更长。结论血清CRP、Glu水平监测可作为颅脑损伤严重程度评价的指标,动态观察颅脑外伤患者血清CRP、Glu水平对疗效观察和预后判断具有重要价值。     

急性脑损伤时内脂素和超敏C-反应蛋白的变化

目的 检测急性颅脑损伤后患者外周血内脂素和超敏C-反应蛋白(hs-CRP)水平的动态变化,探讨其临床意义.方法 120例急性脑损伤患者均为滕州市中心人民医院神经外科及ICU科2009年8月至2010年6月收治住院的患者,其中男60例,女 60例,年龄(43.2±6.2)岁.所有患者均符合颅脑损伤诊断标准.根据入院时格拉斯哥评分(GCS)将患者分为轻度(13～15分)40例,中度(9～12分)40例,重度(3～8分)40例.60例健康体检人员作为对照,其中男30例,女30例,年龄(42.2±6.7)岁.患者于入院后12 h内(即0时间点)及1,3,7,15 d空腹抽取静脉血3 mL用于检测外周血hs-CRP和内脂素的含量变化.结果 急性颅脑损伤患者血清hs-CRP水平均显著高于正常人对照组(3.05±2.08)mg/L(P＜0.01),且患者hs-CRP水平与病情呈正相关;急性颅脑损伤患者血清内脂素含量明显高于正常对照组(17.61±2.45)μg/L(P＜0.01),与病情呈正相关;各组患者在入院当天hs-CRP水平有所升高,并在入院后继续升高,hs-CRP在1 d达高峰,3 d开始下降;内脂素含量在入院当天有所升高,3 d达到高峰,以后逐渐上升;患者内脂素水平与hs-CRP呈正相关(r=0.63,P＜0.01).结论 急性颅脑损伤患者外周血内脂素和hs-CRP水平的变化与病情严重程度密切相关.