戊四氮致痫幼鼠不同脑区脑损伤研究

目的：研究S100B蛋白、碱性髓鞘蛋白（MBP）和神经元特异性烯醇酶（NSE）在戊四氮（PTZ）诱导致痫幼鼠海马、额叶、中脑和血清中的动态改变,评鉴癫痫（EP）造成不同脑区的神经损伤。方法雄性SD幼鼠60只,随机分对照组（10只）和实验组（50只）。实验组腹腔注射（IP）PTZ 50 mg/kg 1次,A组IP生理盐水。根据EP发作分级,0～1级9只视为B组,立即取脑;2级以上发作41只,于EP发作后0 h（C＝10）、6 h（D＝11）、24 h （E＝10）、72 h（F＝10）断头,分别取下海马、中脑和额叶皮质,断头前抽取躯干血。采用ELISA法检测血清和各脑区S100B和MBP ,采用EIA法检测血清和各脑区 NSE值。结果 EP发作后,海马、额叶和血清S100B蛋白、MBP和NSE均明显高于对照组。海马和额叶的NSE峰值出现在中脑NSE之前。各脑区和血清S100B蛋白皆于EP后24 h达高峰,而MBP于EP后72 h达高峰。结论 EP发作可以造成大脑不同脑区神经元细胞、胶质细胞和神经髓鞘的损伤。     

热性惊厥患儿烯醇化酶与脑电图检查的临床研究

目的测定热性惊厥(FS)患儿血清中神经元特异性烯醇化酶(NSE)水平与脑电图(EEG)变化,观察NSE与脑损伤的关系,以便指导临床治疗和改善预后。方法选择FS组患儿208例,其中单纯性热性惊厥(SFS)166例,复杂性热性惊厥(CFS)42例,对照组为112名同期我院健康查体儿童。FS组患儿于惊厥发作后24 h内抽取静脉血采用电化学发光法检测血清NSE,对照组标本留取及处理方法相同。FS组患儿分别于发病后第1天和第14天进行EEG检查。结果 FS组患儿血清中NSE水平明显高于健康对照组(P<0.01);CFS组与健康对照组和SFS组比较差异均具有统计学意义(P<0.01)。FS组患儿惊厥发作后第1天EEG异常率为79.8%,其中SFS组78.3%,CFS组85.7%,差异无统计学意义(P>0.05);第14天EEG异常率为24.0%,SFS组仅16.3%,CFS组则54.8%,2组比较差异具有统计学意义(P<0.01)。结论 FS后及时测定血清NSE水平,并结合EEG检查对判断脑损伤程度及预后具有重要价值。     

延胡索酸水合酶缺陷型平滑肌瘤的研究进展

延胡索酸水合酶（FH）缺陷型平滑肌瘤是子宫平滑肌瘤的一种罕见病理类型,它是FH基因胚系或体系突变引起,且以后者居多。该病虽较为罕见,但当它作为遗传性平滑肌瘤病和肾细胞癌（HLRCC）综合征的前哨表现时,严重的症状体征导致的早期手术以及术后病理为HLRCC综合征提供了早期诊断的可能。FH缺陷型平滑肌瘤的诊断以系统化的诊断方法为主,形态学和免疫组织化学染色有助于提高检出率,但价值有限。虽然FH缺陷型平滑肌瘤的生物学潜能低,但突变来源不同导致患者治疗和预后也大不相同。因此对于临床医师而言,散发性和遗传性FH缺陷型子宫肌瘤的鉴别是一个巨大挑战。该文就FH缺陷型子宫平滑肌瘤的临床表现、形态学特征、诊断与鉴别诊断、治疗及预后做一综述。     

Polyunsaturated fatty acid saturation by gut lactic acid bacteria affecting host lipid composition

In the representative gut bacterium Lactobacillus plantarum, we identified genes encoding the enzymes involved in a saturation metabolism of polyunsaturated fatty acids and revealed in detail the metabolic pathway that generates hydroxy fatty acids, oxo fatty acids, conjugated fatty acids, and partially saturated trans-fatty acids as intermediates. Furthermore, we observed these intermediates, especially hydroxy fatty acids, in host organs. Levels of hydroxy fatty acids were much higher in specific pathogen-free mice than in germ-free mice, indicating that these fatty acids are generated through polyunsaturated fatty acids metabolism of gastrointestinal microorganisms. These findings suggested that lipid metabolism by gastrointestinal microbes affects the health of the host by modifying fatty acid composition.Dietary fats are metabolized not only by humans but also by microbes in our gastrointestinal tracts. Microorganisms in the gastrointestinal tract interact with their host in many ways and contribute significantly to the maintenance of host health (1). Lipid metabolism by gastrointestinal microbes generates multiple fatty acid species, such as conjugated fatty acids and trans-fatty acids, that can affect host lipid metabolism (2). However, lipid metabolism by gastrointestinal microbes has not been explored in detail. Saturation metabolism of polyunsaturated fatty acids, a representative mode of lipid metabolism by gastrointestinal microbes, is a detoxifying metabolism of anaerobic bacteria, such as lactic acid bacteria, that reside in colon and intestine. This process transforms growth-inhibiting free polyunsaturated fatty acids into less toxic free saturated fatty acids (3). This saturation metabolism generates characteristic fatty acids (e.g., conjugated fatty acids and trans-fatty acids, which are well known to present in ruminant-derived foods and exert various physiological activities).“Conjugated fatty acid” is a collective term for positional and geometric isomers of fatty acids with conjugated double bonds. In particular, conjugated linoleic acids (CLAs), such as cis-9,trans-11-CLA and trans-10,cis-12-CLA, reduce carcinogenesis (4), atherosclerosis (5), and body fat (6). With regard to lipid metabolism, CLA is a potent peroxisome proliferator-activated receptor (PPAR)α agonist (7), and treatment with CLA increases the catabolism of lipids in the liver of rodents (8). Based on these findings, CLA is now commercialized as a functional food for control of body weight, especially in the United States and European countries.On the other hand, consumption of trans-fatty acids increases the risk of coronary heart disease by increasing LDL and reducing HDL cholesterol levels (9). Consequently, trans-fatty acids are considered to be harmful for health, and nutritional authorities have recommended that consumption of trans-fatty acids be reduced to trace amounts (10). Therefore, it is important to control fatty acid saturation processes that generate these fatty acids (11); however, the precise metabolic pathway and enzymes involved have not been clearly identified.Our analyses on conjugated fatty acid synthesis in representative gut bacteria, the lactic acid bacteria (12–15), demonstrated that Lactobacillus plantarum AKU 1009a (AKU Culture Collection, Faculty of Agriculture, Kyoto University) can transform the cis-9,cis-12 diene structure of C18 fatty acids such as linoleic acid, α-linolenic acid, and γ-linolenic acid into the conjugated diene structures cis-9,trans-11 and trans-9,trans-11 (16–21). In addition, this strain can saturate these conjugated dienes into the trans-10 monoene. Our subsequent metabolic analysis indicated that 10-hydroxy-12-octadecenoic acid is an intermediate of CLA synthesis, and further investigations of hydroxy fatty acid metabolism by lactic acid bacteria revealed that CLA is produced from hydroxy fatty acids such as ricinoleic acid in castor oil (22–25). In cell-free extracts from this strain, we identified the enzymes involved in CLA synthesis (26). Three enzymes, CLA-HY, CLA-DH, and CLA-DC, are necessary for synthesis of conjugated fatty acids such as CLA. Only the combined action of these three enzymes can generate CLA from linoleic acid, with 10-hydroxy-cis-12-octadecenoic acid arising as an intermediate (Fig. 1B, 3). The reactions catalyzed by each enzyme, however, were not revealed in those studies. Through genomic analysis in L. plantarum WCFS1, we found that cla-dh (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"NC_004567","term_id":"380031102","term_text":"NC_004567"}}NC_004567; region: 59613-60473) and cla-dc (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"NC_004567","term_id":"380031102","term_text":"NC_004567"}}NC_004567; region: 60505-61350) are located in a cluster with another gene, cla-er (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"NC_004567","term_id":"380031102","term_text":"NC_004567"}}NC_004567; region: 61378-62031) (Fig. 1A). In light of this, we tried to identify the function of the gene product (CLA-ER) together with those of CLA-HY, CLA-DH, and CLA-DC.Open in a separate windowFig. 1.Gene clusters for polyunsaturated fatty acid metabolism and GC chromatograms. (A) Gene clusters for fatty acid metabolic enzymes in L. plantarum. (B) GC chromatograms of substrate (1); reaction with CLA-HY (2); reaction with CLA-HY, CLA-DH, and CLA-DC together with FAD and NADH (3); and reaction with CLA-HY, CLA-DH, CLA-DC, and CLA-ER together with FAD and NADH (4). I.S., internal standard.     

间断性脐带闭塞对胎羊静脉导管搏动指数、Tei指数、脐血神经元特异性烯醇化酶、S100B蛋白的影响及其相关性分析

目的 探讨胎羊间断性脐带闭塞后静脉导管搏动指数（ductus venosus pulsatility index，DVPI）、Tel指数评估胎儿缺氧缺血性脑损伤的应用价值。方法 7只晚孕山羊作为实验组，于妊娠116～130d对胎羊行宫内外科手术，用套囊充气闭合器完全夹闭脐带，每次夹闭90S，间隔30min，重复5次。于实验前5min、每次夹闭后3min应用彩色多普勒血流显像技术检测DVPI、Tei指数，抽取胎羊脐动脉血做血气分析，应用电化学发光法测定脐血神经元特异性烯醇化酶（neuron specific enolase，NSE）、S100B蛋白含量，同时分析这些指标的相关性。另选5只晚孕山羊作为对照组，只手术不夹闭脐带。结果 实验组胎羊脐动脉血PaO2和pH随夹闭时间下降，PaCO2、DVPI、右心室和左心室Tei指数、脐血NSE和S100B均随夹闭时间而升高（P均〈0．05）。实验组夹闭后PaO2低于对照组，夹闭后36min开始pH值低于对照组，PaCO2高于对照组；从夹闭后36min开始实验组NSE和S100B蛋白较对照组升高（P均〈0．05）。实验组DVPI、右心室Tei指数、左心室Tei指数、脐血NSE、S100B含量与血pH值呈负相关（r分别为-0．358、-0．538，-0．681、-0．388、-0．539，P均〈0．05），与血PaO2呈负相关（r分别为-0．435、-0．684、-0．725、-0．602，-0．447，P均〈0．01），与血PaCO2呈正相关（r分别为0．699、0．682、0．780、0．477、0．527，P均〈0．01）。实验组DVPI、右心室和左心室Tei指数与血清NSE含量呈正相关（r分别为0．394、0．653、0．687，P均〈0．05），与血清S100B含量呈正相关（r分别为0．359、0．606、0．640，P均〈0．05）。结论 Tei指数与血气指标及血清NSE、S100B的相关性较好，可能成为预测胎儿缺氧缺血性脑损伤简便而有效的指标。     

癫痫病儿血清神经元特异性烯醇化酶水平的检测

①目的　探讨癫痫病儿发作后不同时间血清神经元特异性烯醇化酶 (NSE)的动态变化。②方法采用放射免疫分析法测定 11例癫痫病儿发作后 4 8h内、2～ 4d、5～ 7d血清中NSE水平 ,并与 2 2例正常儿童进行对照。③结果　癫痫病儿发作 4 8h内及 2～ 4d血清NSE水平与对照组比较 ,差异有显著性 (t =4 .32、2 .5 8,P<0 .0 1、0 .0 5 )。④结论　癫痫发作后血清NSE水平升高可能与脑神经元损害有关。     

异丙酚预处理对全脑缺血再灌注大鼠脑组织S100β和神经元特异性烯醇化酶的影响

目的探讨异丙酚预处理对全脑缺血再灌注大鼠脑组织S100β及神经元特异性烯醇化酶（NSE）含量的影响，评价异丙酚对脑组织的保护作用。方法30只雄性SD大鼠，随机分为假手术组（A组）、单纯脑缺血再灌注组（B组）和缺血前2h异丙酚预处理组（C组），每组10只，C组动物在缺血前腹腔注射异丙酚100mg／kg，采用线栓法制作全脑缺血再灌注模型，于脑缺血再灌注后24h评估并记录动物的神经行为学评分，测定大鼠脑组织S100β和NSE含量。结果异丙酚预处理组神经行为学评分较单纯脑缺血再灌注组高（P〈0．05），假手术组评分也明显高于单纯脑缺血再灌注组（P〈0．05），异丙酚预处理组脑组织中S100β和NSE含量明显低于单纯脑缺血再灌注组（P〈0．05）。结论异丙酚可以降低脑缺血再灌注损伤大鼠脑组织S100β和NSE的含量，减轻神经损害，缺血前2h异丙酚预处理可以改善大鼠脑缺血再灌注损伤后的神经行为学评分，有明显的脑保护作用。     

脑缺血再灌注恢复期大鼠梗死周围组织GFAP、NSE、SYN、Nogo—A表达与神经功能转归的相关性

目的探讨脑缺血再灌注恢复期大鼠梗死周围组织胶质纤维酸性蛋白（glial fibrillary acidic protein,GFAP）、神经元特异性烯醇化酶（neuron—specific enolase,NSE）、突触素（synaptophysin,SYN）、神经突生长抑制因子-A（neurite outgrowth inhibitor—A,Nogo—A）表达与神经功能转归的相关性。方法线栓法制作大脑中动脉闭塞（middle cerebral artery occlusion,MCAO）2h再灌注模型,模型制作后28、35、42和49d改良神经功能缺损评分（modified neurological severity score,mNSS）,并采用免疫组化方法检测脑梗死周围组织GFAP、NSE、SYN、Nogo—A表达。结果大鼠脑缺血再灌注后随着时间推移,mNSS评分逐渐下降,除第35天（5．11±0．737）与第42天（4．54±0．519）、第42天与第49天（4．29±0．488）无显著差异外,其余各时间点均有显著差异（P均〈0．05）。GFAP阳性细胞数量从第28天至第49天逐渐减少,其中第42天（51．00±13．59）和第49天（44．38±11．94）显著少于第28天（69．00±15．10）（P〈0．05）。NSE表达的累积光密度（integrated optical density,IOD）逐渐增高,第28天（6218．57±1864．25）与第42（9414．00±2491．12）和第49天（12522．50±3106．99）,第35天（7343．40±1533．35）与第49天差异显著（P均〈0．05）,其余各时间点无显著差异。SYN表达IOD逐渐增高,第49天（66503．00±12834．61）显著高于第28天（43905．14±13208．59）（P〈0．05）。Nogo-A阳性细胞数量逐渐减少,第49天（42．13±14．45）显著少于第28天（59．57±15．25）（P〈0．05）。GFAP表达与mNSS评分呈正相关（r=0．993,P=0．007）,NSE（r=～0．954,P=0．044）、SYN（r=-0．992,P=0．008）表达与mNSS评分呈负相关。结论大鼠脑缺血再灌注后恢复期神经功能转归与GFAP表达下调、NSE和SYN表达上调相关。     

肠道病毒中枢神经系统感染病儿免疫功能紊乱与神经元损害的相关性

目的通过检测肠道病毒（EV）中枢神经系统感染病儿脑脊液中白细胞介素-10（IL-10）、γ-干扰素（IFN-γ）及神经元特异性烯醇化酶（NSE）的水平,探讨EV引起病儿免疫功能紊乱的机制。方法收集2008年1月—2009年1月青岛大学医学院附属医院和滨州医学院附属医院临床诊断为无菌性脑膜炎、脑炎病儿的急性期脑脊液标本100例。采用逆转录聚合酶链反应（RT-PCR）方法,检测标本中的EV RNA;应用双抗体夹心ELISA法检测EV感染组和对照组脑脊液中IL-10、IFN-γ及NSE的表达水平。结果 100例急性期病儿脑脊液中,经RT-PCR和二次PCR共获得阳性结果78例（78.0%）;EV感染组IFN-γ的表达水平高于对照组（t=4.24,P〈0.01）,重症组高于轻症组（t=4.13,P〈0.01）;IL-10表达水平低于对照组（t=7.89,P〈0.01）,重症组低于轻症组（t=2.97,P〈0.01）;NSE表达水平高于对照组（t=7.02,P〈0.01）,重症组高于轻症组（t=2.79,P〈0.01）。EV感染组NSE与IFN-γ水平呈正相关（r=0.673,P〈0.05）,与IL-10水平呈负相关（r=-0.587,P〈0.05）。结论 IL-10和IFN-γ参与EV对中枢神经系统免疫损害的过程,并与神经元的损害程度密切相关。