中药脑溢安改善脑出血大鼠神经元有氧化谢的药效机制

目的 通过观察脑出血大鼠诱导型一氧化氮合酶（inducible nitric oxide synthase,iNOS）在星形胶质细胞中的表达及中药脑溢安的影响，探讨脑出血的机制和中药的脑保护作用。方法 100只动物按数字表法随机分成假手术组（25只）、手术组（75只）。其中手术组造模后又随机分入模型组、脑溢安治疗组、AG治疗组，每组25只。采用胶原酶复制大鼠脑出血模型，以免疫荧光双标组化法和蛋白质印迹法观察脑出血后iNOS的表达情况。结果 脑出血后，模型组大鼠表达iNOS的反应性星形胶质细胞主要是分布在基底核血肿上部周边；蛋白印迹结果明显，模型组大鼠基底核区iNOS从12h，1d阳性信号逐渐增强（t=15.3，P&;lt;0.01），4d达峰值（t=36.4，P&;lt;0.01），7d信号稍弱（t=27.73，P&;lt;0.01）；脑溢安组和氢基胍组的阳性信号较前者弱。结论 脑出血后，患侧基底核反应性星形胶质细胞中的iNOS表达增强；脑溢安可能通过抑制星形胶质细胞中的iNOS表达，实现其脑保护作用。     

中药对脑出血后含铁血红素氧合酶-1蛋白表达的影响

目的:研究脑出血后脑内含铁血红素氧合酶-1(HO-1)蛋白的表达以及中药脑溢安的干预作用.方法:用Ⅶ型胶原酶立体定位注入法制备脑出血大鼠模型.100只大鼠随机分为假手术组(20只)、模型组(40只)和脑溢安组(40只,脑溢安4.92 g·kg-1·d-1).各组分别于术后12、24 h及2、4、7 d用免疫荧光双标技术检测脑组织HO1的表达部位和表达细胞种类;用蛋白质免疫印迹法(Western blot)评价HO1蛋白的表达变化.结果:HO-1免疫阳性细胞主要分布在模型组动物患侧基底节,以小胶质细胞为主.Western blot显示:脑出血后HO-1蛋白在12 h即有表达,2 d达高峰,而后逐渐减弱;脑溢安治疗组的表达趋势与模型组相同,2 d时表达水平明显高于以后的时间点.结论:中药脑溢安可促进HO-1的表达,这可能是其通过活血化瘀治疗脑出血、实现脑保护的机制之一.     

安脑丸对急性脑出血大鼠OX42、脑源性神经营养因子及突触素表达的影响

目的:观察安脑丸对大鼠脑出血后OX42、脑源性神经营养因子(BDNF)及突触素(SYN)表达的影响。方法:SD大鼠190只随机分为正常组10只,假手术组、模型组、安脑丸组各60只;后3组又随机分为造模后12h、1d、2d、4d、7d、10d共6个时间点,各10只。模型组制作脑出血模型,安脑丸组加用安脑丸灌胃。于各时间点采用免疫组化法及western blot法检测OX42、BDNF及SYN表达情况。结果:模型组及安脑丸组OX42及BDNF阳性细胞数及SYN、BDNF蛋白表达显著高于正常组及假手术组(P<0.01),安脑丸组OX42阳性细胞数显著少于模型组(P<0.01),BDNF阳性细胞数及SYN、BDNF蛋白表达显著高于模型组(P<0.01)。结论:小胶质细胞的激活、BDNF及SYN的表达与脑出血密切相关,安脑丸可显著减少脑出血后小胶质细胞的活化,增加BDNF及SYN的表达。     

脑出血大鼠应用脑溢安后脑内多唾液酸神经细胞黏附分子的表达

目的:多唾液酸神经细胞黏附分子是发育阶段神经干细胞膜上特异表达的糖蛋白,观察脑出血后其在脑内的表达与中药脑溢安干预的关系。方法:实验于2004-09/11在中南大学湘雅医院中西医结合科实验室完成。125只SD大鼠随机分为正常对照组5只,假手术组40只,模型组40只,脑溢安组40只。模型组和脑溢安组用Ⅶ型胶原酶诱导大鼠脑出血建立模型,假手术组用生理盐水代替Ⅶ型胶原酶,正常对照组不造模。给药:正常对照组与模型组不给药,假手术组颅内注射2μL生理盐水,脑溢安组行脑溢安浸膏(主要含钩藤、大黄、丹皮、地龙等)灌胃,2mL/次,2次/d。假手术组、模型组、脑溢安组分别于术后1,3,5,7,10,14,21,28d8个时间点将大鼠麻醉后分批处死5只,取脑组织切片进行多唾液酸神经细胞黏附分子的免疫组化观察,以阳性细胞数作为观察指标。结果:125只大鼠全部进入结果分析。①正常对照组和假手术组术后1,3,5,7,10,14,21,28d均无多唾液酸神经细胞黏附分子阳性表达。②脑溢安组及模型组大鼠均于术后5d在血肿区出现多唾液酸神经细胞黏附分子阳性表达,7d达高峰,10d开始下降,至术后28d多唾液酸神经细胞黏附分子阳性显色逐渐减少,并完全集中在血肿近侧脑室及海马处,两组变化趋势相同,阳性表达均高于正常对照组。③术后14d脑溢安组多唾液酸神经细胞黏附分子阳性细胞计数开始多于模型组(t=-2.984,P=0.017),至28d仍显著高于模型组(t=-3.99,P=0.004)。结论:脑出血大鼠术后5d血肿区出现多唾液酸神经细胞黏附分子,呈阳性表达,证明其脑内存在神经干细胞重塑。通过强化此种重塑可能是脑溢安促进脑出血后神经功能恢复的途径之一。     

脑溢安颗粒对实验性大鼠脑出血后神经干细胞增殖的影响

目的:探讨中药脑溢安颗粒对脑出血后神经干细胞反应的影响。方法:72只雄性SD大鼠单纯随机分为假手术组、模型组、脑溢安组,采用Rosenberg方法复制大鼠脑出血模型,分别给予不同处理,于术后1,12h,1,3,5,7d处死动物,运用免疫组织化学,Westernblot方法检测脑出血后各组的nestin表达。结果:脑出血后,nestin阳性细胞增多,1d达高峰,然后逐渐降低,维持到第7天,nestin阳性细胞主要分布在海马、皮质区,且脑溢安组与模型组1d前各时间点差异无显著性意义(P>0.05),3d后各时间点差异有显著性意义(t分别为3.081,8.034,5.833,P<0.05～0.01);nestin蛋白表达有相同趋势。结论:正常脑组织中存在一定数量的神经干细胞,脑出血后神经干细胞被激活增殖,中药脑溢安能维持促进增殖,这种作用可能是通过调节细胞外因子来实现。     

脑出血大鼠应用脑溢安后脑内多唾液酸神经细胞黏附分子的表达

目的：多唾液酸神经细胞黏附分子是发育阶段神经干细胞膜上特异表达的糖蛋白，观察脑出血后其在脑内的表达与中药脑溢安干预的关系。方法：实验于2004—09／11在中南大学湘雅医院中西医结合科实验室完成。125只SD大鼠随机分为正常对照组5只，假手术组40只，模型组40只，脑溢安组40只。模型组和脑溢安组用Ⅶ型胶原酶诱导大鼠脑出血建立模型，假手术组用生理盐水代替Ⅶ型胶原酶，正常对照组不造模。给药：正常对照组与模型组不给药，假手术组颅内注射2μL生理盐水，脑溢安组行脑溢安浸膏（主要含钩藤、大黄、丹皮、地龙等）灌胃，2mL／次，2次／d。假手术组、模型组、脑溢安组分别于术后1，3，5，7，10，14，21，28d8个时间点将大鼠麻醉后分批处死5只，取脑组织切片进行多唾液酸神经细胞黏附分子的免疫组化观察，以阳性细胞数作为观察指标。结果：125只大鼠全部进入结果分析。①正常对照组和假手术组术后1，3，5，7，10，14，21，28d均无多唾液酸神经细胞黏附分子阳性表达。②脑溢安组及模型组大鼠均于术后5d在血肿区出现多唾液酸神经细胞黏附分子阳性表达，7d达高峰，10d开始下降，至术后28d多唾液酸神经细胞黏附分子阳性显色逐渐减少，并完全集中在血肿近侧脑室及海马处，两组变化趋势相同，阳性表达均高于正常对照组。③术后14d脑溢安组多唾液酸神经细胞黏附分子阳性细胞计数开始多于模型组（t=2．984，P=0．017），至28d仍显著高于模型组（t=-3．99，P=0．004）。结论：脑出血大鼠术后5d血肿区出现多唾液酸神经细胞黏附分子，呈阳性表达，证明其脑内存在神经干细胞重塑。通过强化此种重塑可能是脑溢安促进脑出血后神经功能恢复的途径之一。     

脑缺血上调大鼠神经元和星形胶质细胞CDK4的表达

目的:研究大鼠局灶性脑缺血再灌注损伤后CDK4在神经元和星形胶质细胞的表达。方法:SD大鼠25只,随机分为假手术组和再灌注1d、3d、7d、14d组,建立大鼠大脑中动脉阻塞(MCAO)再灌注模型,假手术组仅进行手术不造成缺血状态,应用流式细胞术检测各组神经元和星形胶质细胞中CDK4的表达。结果:缺血侧皮质星形胶质细胞和神经元中的CDK4的表达在再灌注7d、14d后表达上调,与假手术组比较有差异(P<0.05);大脑皮质神经元和星形胶质细胞的比较发现,神经元中的CDK4在假手术组、再灌注7d组的表达水平高于星形胶质细胞(P<0.05)。结论:脑缺血后,缺血侧皮质区星形胶质细胞和神经元的CDK4表达上调。     

脑溢安颗粒对实验性大鼠脑出血后神经干细胞增殖的影响

目的：探讨中药脑溢安颗粒对脑出血后神经干细胞反应的影响。方法：72只雄性SD大鼠单纯随机分为假手术组、模型组、脑溢安组。采用Roeenberg方法复制大鼠脑出血模型。分别给予不同处理。于术后1，12h，1，3，5，7d处死动物。运用免疫组织化学。Western blot方法检测脑出血后各组的nestin表达。结果：脑出血后，nestin阳性细胞增多，ld达高峰，然后逐渐降低，维持到第7天，nestin阳性细胞主要分布在海马、皮质区，且脑溢安组与模型组1d前各时间点差异无显著性意义(P&;gt;0．05)。3d后各时间点差异有显著性意义(t分别为3．081，8．034，5．833,P&;lt;0．05～0.01)；nestin蛋白表达有相同趋势。结论：正常脑组织中存在一定数量的神经干细胞，脑出血后神经干细胞被激活增殖。中药脑溢安能维持促进增殖，这种作用可能是通过调节细胞外因子来实现。     

脑溢安颗粒对脑出血大鼠脑组织病理改变及一氧化氮生成的影响

目的探讨脑溢安颗粒(简称脑溢安)治疗脑出血的作用机制.方法立体定位内囊注射胶原酶(Ⅶ型)诱导大鼠脑出血模型,Griess法检测血肿周围大脑皮质内一氧化氮(nitricoxide,NO)含量,光镜观察脑组织病理改变.结果模型组大鼠大脑皮质NO含量显著高于手术对照组(P<0.01).脑溢安组大鼠大脑皮质NO含量较模型组相应时间点明显降低(P<0.01).结论脑溢安可能通过减少脑内NO产生,对脑出血损伤发挥保护作用.     

实验性脑出血6个月内神经行为学和神经胶质细胞的变化

目的观察实验性脑卒中恢复期的神经行为学得分、患侧病灶周围以及健侧相应区域的神经胶质细胞的变化情况,为研究脑出血后遗症期工作打基础。方法用高血压大鼠制作脑出血模型,根据Bederson评定方法记录脑出血后1,3,6个月的神经行为学改变得分;同时在640倍镜下,在患侧脑病灶周围选5个视野对病灶周围的神经胶质细胞进行计数,然后算每个视野的均数和标准差;在健侧脑的同样部位也进行同样的计数和统计。结果(1)3个月内,正常血压脑出血组神经行为学得分从7.2±0.7下降到6.3±1.5(t=3.0689,P<0.01),6个月时下降到5.8±1.8(t=4.0982,P<0.01);高血压脑出血组神经行为学得分3个月内从8.5±0.7下降到7.3±0.9(t=3.5968,P<0.01),6个月时下降到6.1±0.5(t=8.0134,P<0.01)。(2)所有脑出血组术后6个月出现脑萎缩和脑皮质变薄。(3)所有脑出血组1,3,6个月时病灶周围及健侧相应区域的胶质细胞数量均较术前大量增加(P<0.01),且以3个月时为高峰。(4)神经行为学改变与患侧、健侧胶质细胞数变化之间存在明显相关性联系r=0.9879(P<0.01)和r=0.734(P<0.01)。结论(1)脑出血后确实存在着自然恢复的现象。但同时出现的脑萎缩,脑皮质变薄的现象提示似乎神经行为学的改善与脑皮质的功能关系并不大。(2)6个月内病灶周围的胶质细胞的增     

Determination of creatine kinase isoenzyme MB activity in serum using immunological inhibition of creatine kinase M subunit activity. Activity kinetics and diagnostic significance in myocardial infarction.

This is a new method for the determination of creatine kinase isoenzyme MB activity in serum. The method uses direct activity measurement of creatine kinase B subunit activity after blocking of CK-M subunit activity by inhibiting antibodies. The test takes no longer than 15 min. The method yields an intra-serial C.V. of 2.0-12.9%, and a C.V. from day to day of 5.5%. The detection limit is 3.4 U/l creatine kinase MB. In the 95 cases with proven myocardial infarction several types of creatine kinase MB activity kinetics could be determined. The percentage of creatine kinase MB of peak CK-total is 6-25%, with a mean of 11.1%. The amount of creatine kinase MB with respect to total CK activity after reinfarction is higher than the amount after initial infarction.     

Dissociable correlates of two classes of retrieval processing in prefrontal cortex

Although substantial evidence suggests that the prefrontal cortex (PFC) implements processes that are critical for accurate episodic memory judgments, the specific roles of different PFC subregions remain unclear. Here, we used event-related functional magnetic resonance imaging to distinguish between prefrontal activity related to operations that (1) influence processing of retrieval cues based on current task demands, or (2) are involved in monitoring the outputs of retrieval. Fourteen participants studied auditory words spoken by a male or female speaker and completed memory tests in which the stimuli were unstudied foil words and studied words spoken by either the same speaker at study, or the alternate speaker. On "general" test trials, participants were to determine whether each word was studied, regardless of the voice of the speaker, whereas on "specific" test trials, participants were to additionally distinguish between studied words that were spoken in the same voice or a different voice at study. Thus, on specific test trials, participants were explicitly required to attend to voice information in order to evaluate each test item. Anterior (right BA 10), dorsolateral prefrontal (right BA 46), and inferior frontal (bilateral BA 47/12) regions were more active during specific than during general trials. Activation in anterior and dorsolateral PFC was enhanced during specific test trials even in response to unstudied items, suggesting that activation in these regions was related to the differential processing of retrieval cues in the two tasks. In contrast, differences between specific and general test trials in inferior frontal regions (bilateral BA 47/12) were seen only for studied items, suggesting a role for these regions in post-retrieval monitoring processes. Results from this study are consistent with the idea that different PFC subregions implement distinct, but complementary processes that collectively support accurate episodic memory judgments.     

俯卧位通气对高海拔地区肺复张治疗无效急性呼吸窘迫综合征患者氧合的影响

目的 探讨俯卧位通气对高海拔地区肺复张术(RM)治疗无效急性呼吸窘迫综合征(ARDS)患者的治疗作用.方法 从海拔2260m的地区医院筛选RM治疗无效的41例ARDS患者[平均氧合指数( PaO2/FiO2)较RM前升高＜20％视为RM无效],依不同病因分为肺内源性ARDS组(ARDSp组)和肺外源性ARDS组(ARDSexp组),每组再按信封法随机分为俯卧位组和仰卧位组,即ARDSp俯卧位组(11例)、ARDSp仰卧位组(9例)、ARDSexp俯卧位组(10例)、ARDSexp仰卧位组(11例).在通气前及通气1、2、3、4h监测动脉血氧分压( PaO2)、PaO2/FiO2、静态顺应性(Cst)、气道阻力(Raw)的变化.结果 通气lh时,ARDSexp俯卧位组PaO2/FiO2( mm Hg,l mm Hg=0.133 kPa)即较通气前显著升高(157.4±40.6比129.3±48.7,P＜0.05),并随通气时间延长呈持续增高趋势,4h达峰值(219.1 ±41.1);且ARDSexp俯卧位组通气3h内PaO2/FiO2较其他3组显著增高,另3组间则差异无统计学意义.ARDSp俯卧位组、ARDSexp俯卧位组通气4h时PaO2/FiO2均较相应仰卧位组显著增高(208.8±39.7比127.4±47.1,219.1±41.1比124.9±50.8,均P＜0.05).4组通气前后Cst无显著改变,各组间差异也无统计学意义.ARDSp俯卧位组通气4h时Raw(cmH2O·L-1·s-1)较通气前显著降低(6.8±1.7比10.7±1.8,P＜0.05),且明显低于其他3组;其他3组各时间点Raw组内及组间比较差异均无统计学意义.结论 俯卧位通气作为ARDS机械通气重要策略之一,可以改善RM无效高原ARDS患者的氧合,为抢救患者赢得宝贵的时间.     

Digital imaging among medical facilities

The Department of Veterans Affairs (VA) in the USA operates a network of 172 medical centres which all utilize a hospital information system (HIS) which has been developed and is currently maintained by the VA. During the past several years, an image management and communication module has been developed, installed and clinically utilized at the Washington DC and Maryland VA Medical Centres. This image management and communication system, referred to as the decentralized hospital computer program (DHCP) imaging system, is fully integrated with a commercial picture archiving and communication system (PACS). The system is utilized to capture, archive, and display all images generated within the hospital including radiology, nuclear medicine, pathology, endoscopy, bronchoscopy, and dermatology, intraoperative photographs, ECG data, and a limited number of paper documents. The ultimate goal of the project is to have all patient text and image data available at any clinical workstation to any authorized user anywhere within the network of medical centres. Clinical requirements for an imaging workstation include ease of use, rapid and reliable access to the complete set of patient information, and images which are of acceptable quality to meet the requirements of the user and the subspecialty. Patient confidentiality and data security must be safeguarded at all times. Integration of the images with the remainder of the patient's database was found to be critical to the success of the project. The experience at the Washington and Maryland facilities suggests that an imaging system that is successfully integrated with a hospital information system can provide substantial clinical and economic benefits both within and among medical centres. Clinical acceptance and utilization of the system has been excellent, particularly in diagnostic radiology where DHCP Imaging has been interfaced to a commercial PAC system. Based upon this initial experience, the VA has begun to deploy the system throughout its large network of medical centres.     

Myocardial elastography at both high temporal and spatial resolution for the detection of infarcts

Myocardial elastography is a novel method for noninvasively assessing regional myocardial function, with the advantages of high spatial and temporal resolution and high signal-to-noise ratio (SNR). In this paper, in-vivo experiments were performed in anesthetized normal and infarcted mice (one day after left anterior descending coronary artery [LAD] ligation) using a high-resolution (30 MHz) ultrasound system (Vevo 770, VisualSonics Inc., Toronto, ON, Canada). Radiofrequency (RF) signals of the left ventricle (LV) in longitudinal (long-axis) view and the associated electrocardiogram (ECG) were simultaneously acquired. Using a retrospective ECG gating technique, 2-D full field-of-view RF frames were acquired at an extremely high frame rate (8 kHz) that resulted in high-quality incremental displacement and strain estimation of the myocardium. The incremental results were further accumulated to obtain the cumulative displacements and strains. Two-dimensional and M-mode displacement images and strain images (elastograms), as well as displacement and strain profiles as a function of time, were compared between normal and infarcted mice. Incremental results clearly depicted cardiac events including LV contraction, LV relaxation and isovolumetric phases in both normal and infarcted mice, and also evidently indicated reduced motion and deformation in the infarcted myocardium. The elastograms indicated that the infarcted regions underwent thinning during systole rather than thickening, as in the normal case. The cumulative elastograms were found to have higher elastographic SNR (SNR(e)) than the incremental elastograms (e.g., 10.6 vs. 4.7 in a normal myocardium, and 6.0 vs. 2.4 in an infarcted myocardium). Finally, preliminary statistical results from nine normal (m = 9) and seven infarcted (n = 7) mice indicated the capability of the cumulative strain in differentiating infracted from normal myocardia. In conclusion, myocardial elastography could provide regional strain information at simultaneously high temporal (>/=0.125 ms) and spatial ( approximately 55 microm) resolution as well as high precision ( approximately 0.05 microm displacement). This technique was thus capable of accurately characterizing normal myocardial function throughout an entire cardiac cycle, at the same high resolution, and detecting and localizing myocardial infarction in vivo.     

手转胎头术失败的原因与分娩结局临床分析

目的 探讨手转胎头术失败的原因与分娩结局.方法 选择2008年1月至2010年12月于我院住院分娩的持续性枕横位、枕后位产妇198例,根据行手转胎头术后结果分为成功组126例、失败组72例.比较两组分娩结局,对比分析失败原因.结果 失败组胎儿体质量≥3500 g的发生率[76.4％(55/72)]明显高于成功组[31.7％(40/126)],差异有统计学意义(x2=30.177,P=0.001)、失败组宫缩乏力发生率[58.3％(42/72)]高于成功组[38.1％ (48/126)],差异有统计学意义(x2=7.569,P=0.006)、失败组骨盆临界或轻度狭窄发生率[38.9％ (28/72)]高于成功组[23.8％(30/126)],差异有统计学意义(x2 =5.030,P=0.002)、失败组手转胎头时机不当(宫口开大＜6 cm、胎头位于坐骨棘上及宫口开大8～10 cm、胎头位于坐骨棘下≥2 cm)发生率[61.1％(44/72)]高于成功组[38.9％(49/126)],差异有统计学意义(x2=9.084,P=0.003).失败组母儿并发症(产后出血、产褥病率、胎儿窘迫、新生儿窒息)发生率高于成功组(x2 =9.586,P=0.002、x2=9.334,P=0.002、x2=5.910,P=0.015、x2=5.240,P=0.022)、失败组剖宫产发生率[72.2％(52/72)]明显高于成功组[34.1 ％(43/126),x2=26.641,P=0.001)].结论 手转胎头术能使难产变顺产,降低剖宫产率,减少母儿并发症,但须积极预防、处理导致手转胎头术失败的原因,对矫正失败后继续矫正及试产应慎重.