丹参多酚酸盐及神经肽Y干预后癫痫大鼠海马神经肽Y的表达变化

目的探讨丹参多酚酸盐及神经肽Y（neuropeptide-Y,NPY）干预注射对海马NPY的表达影响研究。方法采用匹鲁卡品（PILO）诱发癫痫模型,随机分为NPY干预组、丹参多酚酸盐干预组、盐水干预组和对照组,观察各组大鼠的行为学改变,用免疫组织化学法标记显示各组大鼠脑内海马NPY的表达变化。结果在海马门区、CA3区、CA1区有NPY阳性细胞表达,生理盐水及丹参多酚酸盐干预组可见颗粒细胞中NPY的异位表达。干预方法不同,海马NPY阳性细胞数表达不同,生理盐水（NS）、丹参多酚酸盐干预组NPY阳性细胞数高于对照组,差异有统计学意义（P〈0.05）;丹参多酚酸盐组高于生理盐水干预组,差异有统计学意义（P〈0.05）。结论 NPY有抗癫痫作用。丹参多酚酸盐可以明显降低急性癫痫痉挛性大鼠的发作等级和发作时间,可能通过NPY的表达增高来发挥治疗作用。     

灵芝孢子粉对癫痫大鼠皮质和海马区谷氨酸、γ-氨基丁酸含量的调节

目的：观察使用灵芝孢子粉后，癫痫点燃大鼠模型皮质和海马区谷氨酸和1一氨基丁酸的含量及神经元形态学改变。 方法：实验于2005-08在佳木斯大学基础医学院完成。选择健康雄性wistar大鼠30只（鼠龄12周），随机分为3组，空白对照组、癫痫模型组、灵芝孢子粉组，每组10只。采用戊四氮腹腔注射制作大鼠慢性点燃模型，癫痫模型组采用戊四氮腹腔注射＋生理盐水灌胃；灵芝孢子粉组采用戊四氮腹腔注射＋灵芝孢子粉灌胃；空白对照组采用生理盐水腹腔注射＋生理盐水灌胃。腹腔注射剂量为35mg／kg，灌胃剂量为150mg／kg，1次／d，持续28d。每次注射后观察大鼠60min，记录癫痫发作情况。惊厥发作评分标准：0级，没有行为上的发作；Ⅰ级，节律性点头或头部颤搐；Ⅱ级，阵挛性咀嚼；Ⅲ级，头部颤搐加前肢阵挛性抽搐；Ⅳ级，袋鼠姿势（上身直立）；Ⅴ级，跌倒；Ⅵ级，强直性惊厥。凡显示连续5次Ⅱ级以上惊厥的大鼠为达到点燃标准。点燃后断头取脑，行免疫组化染色观察各指标含量变化及神经元形态学改变。显微镜观察显示棕黄色颗粒者为阳性。结果：实验过程中，无动物死亡，全部进入结果分析。①灵芝孢子粉组和癫痫模型组所有大鼠均有连续多次的Ⅲ～Ⅴ级癫痫发作，均达到癫痫模型点燃标准。②免疫组织化学染色后，空白对照组大脑皮质谷氨酸免疫反应阳性细胞密集分布于各层，海马谷氨酸免疫反应阳性细胞主要分布在海马回的锥体细胞层和齿状回的颗粒层。谷氨酸阳性神经元胞体呈圆形、椭圆形或不规则形，谷氨酸阳性反应产物为棕黄色，呈细颗粒状。癫痫模型组大脑皮质及海马谷氨酸免疫反应阳性细胞数比空白对照组明显增多，差异具有显著性[（206±41），（96±12）个／张，P〈0．05]；[（164±41），（69±14）个／张，P〈0．05]。灵芝孢子粉组大脑皮质及海马谷氨酸免疫反应阳性细胞比癫痫模型组数减少，差异具有显著性[（108±23），（206±41）个／张，P〈0．05]；[（100±12），（164±41）个／张，P〈0．05]。③免疫组织化学染色后空白对照组大脑皮质γ-氨基丁酸有较多分布，浅层尤为明显。大鼠海马回和齿状回均可见γ-氨基丁酸阳性细胞散在分布γ-氨基丁酸阳性神经元胞体呈圆形，胞浆均匀深染。癫痫模型组大脑皮质及海马γ-氨基丁酸免疫反应阳性细胞数比空白对照组明显减少，差异具有显著性[（69±16），（149±21）个／张，P〈0．05]；[（47±9），（108±10）个／张，P〈0．05]；灵芝孢子粉组大脑皮质及海马γ-氨基丁酸免疫反应阳性细胞比癫痫模型组数增多，差异具有显著性[（113±17）（69±16）个／张，P〈0．05]；[（80±11）（47±9），个／张，P〈0．05]。 结论：灵芝孢子粉能够有效降低皮质和海马区兴奋性氨基酸谷氨酸的含量，降低病变神经元兴奋性以达到抗癫痫作用。同时还能增强抑制性氨基酸氨基丁酸的表达，使神经元兴奋性减弱，抑制癫痫的发作，从而减轻癫痫发作给神经系统带来的损伤，所以灵芝孢子粉可能具有减轻痫性发作、保护神经元的作用。     

戊四氮致癫痫大鼠脑组织形态结构及热休克蛋白70表达的初步研究

目的：探讨癫痫的不同发作级别所造成的神经元损伤及热休克蛋白70（HSP70）的表达情况及其意义。方法：采用戊四氮致癫痫模型,随机分为生理盐水对照组和模型组,模型组按癫痫发作级别又分为Ⅰ、Ⅲ、Ⅴ级组。采用常规病理学染色观察各组海马区神经元的受损情况;免疫组织化学方法观察不同级别癫痫发作海马内HSP70表达的变化。结果：Ⅲ、Ⅴ级组与对照组和Ⅰ级组相比,海马结构内存在受损异常神经元,Ⅴ级组更明显（P〈0．05）。同时Ⅲ级组、Ⅴ级组海马各区的HSP70阳性细胞数亦逐渐增多（P〈0．05）。结论：戊四氮致癫痫模型中,随着发作级别的增加,海马结构内神经元损伤逐渐加重,HSP70的表达亦逐渐增加,提示海马结构HSP70的表达与癫痫发作严重程度有关。     

抑制p38/p53信号通路对癫痫大鼠神经元损伤的保护作用

目的 探讨p38/p53信号通路在癫痫持续状态大鼠海马神经元的活化规律,及对癫痫后神经元损伤的保护作用.方法 40只大鼠随机分为正常对照组、癫痫组、抑制剂组、二甲亚枫溶剂对照组,建立氯化锂?匹鲁卡品癫痫大鼠模型,HE染色观察大鼠海马CA1区的神经元病理改变,免疫组化染色观察p38/p?p38、p53/p?p53阳性细胞...     

神经肽Y、丹参多酚酸盐对癫痫大鼠海马FOS蛋白表达的影响及作用探讨

目的探讨NPY及丹参多酚酸盐干预对大鼠海马FOS蛋白的表达影响研究及作用探讨。方法采用匹鲁卡品诱发癫痫状态模型,随机分为丹参多酚酸盐干预组、NPY干预组、盐水干预组和对照组,观察各组大鼠的行为学改变,用免疫组织化学法标记显示各组癫痫发作脑内海马FOS蛋白的表达变化。结果在海马门区、CA3区、CA1区有FOS阳性细胞表达,干预方法不同,海马FOS阳性细胞数表达不同,NS、NPY、丹参多酚酸盐干预组FOS阳性细胞数高于对照组,差异有统计学意义（P〈0.05）。结论丹参多酚酸盐和NPY可能通过某种机制抑制了FOS的表达,在一定程度上起到了抗癫痫作用。     

高功率微波辐射对大鼠脑皮质及海马神经肽Y、nNOS表达的影响

目的：探讨高功率微波（HPM）辐射对大鼠脑皮质和海马形态结构以及神经元神经肽Y（NPY）、神经型-氧化氮合酶（nNOS）表达的影响。方法：分别采用10，30和100mW／cm^2HPM辐射Wistar大鼠，并于微波辐射后6h，1d，3d，7d，14d和28d时处死，同时取出大脑皮质和海马组织。通过HE染色观察实验大鼠脑皮质及海马的形态结构改变情况；采用免疫组化和图像分析技术检测实验大鼠脑皮质和海马神经元NPY、nNOS表达的变化。结果：实验大鼠经10，30，100mW／cm^2HPM辐射后，其脑皮质及海马区神经元固缩、深染；HPM辐射后3d时，大鼠神经元胞浆内nNOS表达显著增加（P〈0.05），NPY表达显著降低（P〈0.01）。结论：10，30及100mW／cm^2HPM辐射可引发大鼠脑皮质和海马神经元损伤，致使其NPY及nNOS表达异常。     

红藻氨酸诱导大鼠癫痫发作时海马神经元凋亡变化的动态观察

目的：探讨大鼠癫痫发作时海马神经元凋亡变化。方法：采用红藻酸氨(KA)诱导大鼠癫痫发作模型，观察行为学及脑电图变化；用神经元尼氏体亚甲蓝特殊染色法观察大鼠癫痫发作时海马CA1区神经元损害情况；用原位细胞凋亡检测法观察癫痫发作时海马CA1区神经元凋亡情况及苯巴比妥干预后神经元凋亡的变化。结果：KA注射后大鼠出现严重边缘性惊厥，脑电图表现为持续性癫痫样放电。海马细胞在KA注射后8h开始出现凋亡阳性细胞，48h达高峰，主要表现在CA1区锥体细胞，经苯巴比妥干预后凋亡细胞明显减少。结论：癫痫发作后的迟发性神经元死亡很可能经由凋亡途径，早期终止癫痫发作可抑制海马神经元凋亡。     

杏仁核点燃癫痫大鼠模型中BKCa通道β4亚基动态变化

目的：观察杏仁核点燃癫痫大鼠BKCa通道β4亚基动态演变情况。方法：建立杏仁核点燃癫痫大鼠模型,随机分对照组、点燃24 h组、30 d组、60 d组。采用尼氏染色观察各组神经元损伤情况;采用实时定量PCR、蛋白质印迹、免疫组织化学检测发作后24 h、30 d、60 d大鼠脑内BKCa通道β4亚基表达的动态改变情况。结果：相比对照组,杏仁核点燃癫痫模型大鼠皮质及海马CA1、CA3区神经元均有明显缺失,且随着点燃时间增加,神经元计数呈下降趋势,各组差异有统计学意义（P<0.05）。点燃后24 h,皮质及海马BKCa通道β4亚基表达下降,且随着点燃时间增加呈下降趋势,差异有统计学意义（P<0.05）。结论：杏仁核点燃可导致脑内神经元持续损伤,同时伴随相应部位BKCa通道β4亚基表达下降,推测β4亚基可能参与了杏仁核点燃癫痫模型发生发展的过程。     

电针穴位刺激对锂-匹罗卡品诱导致痫大鼠行为及脑电活动的影响

目的：观察电针刺激百会穴对锂-匹罗每品诱导致痫大鼠的行为及脑电活动的影响，进一步揭示穴位刺激的抑痫机制。方法：实验于2005—06在解放军第四军医大学西京医院神经内科完成。①选用成年雄性SD大鼠30只。利用脑立体定位手段，将电极埋入大鼠脑部双侧额叶皮质、海马和杏仁核。②选取24只大鼠，脑内植入电极1周后，于大鼠腹腔注射锂-匹罗卡品，造成强直一阵挛性发作癫痫持续状态模型。③将癫痫发作程度（依据Ono等分级标准）Ⅳ级（含Ⅳ级）以上存活大鼠18只随机分为3组；电针穴位刺激组（电针刺激致痫大鼠百会穴），电针刺激对照组（电针刺激致痫大鼠百会穴相临的非穴位处），无刺激对照组（造模后不给予其他干预措施），每组6只。均采用G-6805型电针治疗仪于致痫后第2天开始进行电针刺激干预，其刺激3周。电针刺激参数为：频率为80Hz，电流强度20mA，时间20mn，2次／11。其余6只大鼠为日常对照组：取脑内植入电极，仍未致痫大鼠。腹腔注射与匹罗卡品等量生理盐水。④记录致痫大鼠首次癫痫发作平均潜伏期、注射地西泮前强直一阵挛发作次数和持续时间，以及静默期的长短。此后，每天观察动物的行为学变化4h，记录其静默期后出现的Ono等分级标准Ⅰ～Ⅲ级的自发反复发作次数。⑤采用太阳公司视频脑电描记系统记录脑电。右海马-右耳连续描记各组大鼠致痫前15min到注射地西泮终止发作期间的脑电变化，以及致痫后仵第2，3，4，6周时大鼠的脑电图改变（时间2h）。⑥计量资料差异比较采用方差分析。结果：造模后存活18只及正常大鼠6只进入结果分析。①各组致痫大鼠首次癫痫发作平均潜伏期、注射地西泮前强直一阵挛发作次数和平均持续时间比较，差异不明显（P〉0．05）。电针穴位刺激组大鼠平均每周自发反复发作次数明显少于电针刺激对照组和无刺激对照组大鼠（P〈0．05）。②正常大鼠脑电波形频率以5～10Hz为主，波幅小于200μV。注射后经过潜伏期，大鼠脑电图表现出多种形式的癫痫样波，有单棘渡，多棘波，多相棘波，棘慢波，发作性节律波等。频率最快可达35Hz，波幅高约2．5～3．0mV，发作后可出现抑痫制波。电针穴位刺激组大鼠在第2，3，4，6周时脑区杏仁核内2h内放电次数明显少于电针刺激对照组和无刺激对照组（P〈0．05）。结论：电针刺激百会穴对锂-匹罗卡品诱导致痫大鼠的慢性发作具有明硅的抑痈作用．其电生理机制依赖于抑痫信号的穴位-神经传入解剖途径。     

迷走神经刺激控制海人藻酸诱导大鼠癫痫发作的电生理学研究

目的：通过动物实验，探讨间断性迷走神经刺激(vagus nerve stimulation，VNS)控制癫痫的最佳启动时间及其抗痫效果，观察VNS对海人藻酸致痫大鼠皮质电图和海马神经元电活动的影响。方法：实验选用成年、健康、体质量220—250g的Wistar大鼠，40g／L戊巴比妥钠腹腔麻醉后，颈部皮下注射海人藻酸(10mg／kg)复制癫痫模型。在癫痫发作前后分别给予大鼠左侧迷走神经间断刺激，用银球电极与金属微电极分别记录皮质电图和海马CAl区神经元电活动的变化。结果：①癫痫发作前给予VNS刺激(3.0—4．0mA，33Hz，1ms)50min，皮质电图结果表明可显著降低癫痫发作的严重程度，缩短发作持续时间，甚至可完全抑制发作(F=27．32，P&;lt;0．001)。而癫痫一旦发生再给予上述刺激，则只能降低发作的严重程度，而不能完全阻抑癫痫发作。②当皮下注射海人藻酸诱发海马CAl区神经元出现典型的不规则爆发样电活动后，给予上述刺激，CAl区神经元放电频率有所减少，停止VNS 60min和120min后，放电频率继续显著下降(F=8．531，P&;lt;0．05)。结论：①阻抑海人藻酸致痫模型发作，VNS启动越早，抗痫作用越佳。②间断性VNS对海人藻酸致痫大鼠的抗痫作用。     

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.     

Clinical Pharmacokinetics of Morphine

Morphine, the most widely used mu-opioid analgesic for acute and chronic pain, is the standard against which new analgesics are measured. A thorough understanding of the pharmacokinetics of morphine is required in order to safely and effectively use this analgesic in a wide variety of patients with different levels of organ function. A MEDLINE search was conducted to identify literature published between 1966 and January 2002 relevant to the pharmacokinetics of morphine. These publications were reviewed and the literature summarized regarding unique and clinically important elements of morphine disposition relative to its parenteral administration (including intravenous, intramuscular, subcutaneous, epidural and intrathecal administration), absorption profile (immediate release, controlled release, and sublingual/buccal, and rectal administration), distribution, and its metabolism/ excretion. Special populations, including infants, elderly, and those with renal/liver failure, have a unique morphine pharmacokinetic profile that must be taken into account in order to maximize analgesic efficacy and reduce the risk of adverse events.