改良消减杂交法克隆大鼠脊髓损伤修复相关基因

目的 克隆大鼠脊髓损伤修复相关基因，从分子水平了解中枢神经系统损伤修复的机制。方法 建立大鼠脊髓损伤模型，运用基于聚合酶链反应(PCR)的改良消减杂交技术，克隆与脊髓损伤修复相关的基因。结果 随机选取l10个克隆进行研究，排除假阳性克隆和重复序列后，筛选到47个差异表达序列，其中39个已知序列，8个未知序列。39个已知序列中，包括synuclein、clusterin等与神经系统发育及退行性病变密切相关的蛋白编码序列；8个未知序列中，69号末端含有Leu拉链结构，包含该序列的基因可能编码DNA结合蛋白。结论 运用基于PcR的改良消减杂交技术成功克隆到损伤脊髓上调表达的基因序列，为探讨中枢神经系统损伤修复的分子机制奠定了基础。     

SD大鼠偏侧帕金森病模型差异表达cDNA文库的构建

背景 帕金森病(Parkinson's disease,PD)是好发于中老年人的神经退行性疾病,随着社会的老龄化,其患病率及致残率呈上升趋势.根据流行病学调查,神经病理学研究及基因水平分析,一般认为PD的病因及其病理损伤过程是多因素的,包括遗传因素、环境毒素、氧化应激、兴奋性氨基酸毒性作用、线粒体功能障碍、细胞凋亡和神经营养因子水平降低等.对疾病分子水平变化的研究,不但为阐明疾病的发生发展提供新的资料,而且还能为其治疗开辟新的途径.然而目前对PD分子水平所知尚少,因此,揭示PD分子病理变化,具有十分重要的意义.本研究旨在构建偏侧帕金森病SD大鼠纹状体组织差异表达基因消减cDNA文库,并筛选出一些可能参与PD病理损伤过程的基因,为PD的基因治疗提供依据.方法 行SD大鼠纹状体区立体定向注射6-OHDA制备偏侧帕金森病模型;用抑制消减杂交方法分离差异表达基因的cDNA片段,连接T载体构建文库,转化扩增后随机挑取白色克隆行菌落PCR鉴定;利用反Northern杂交对重组质粒进行筛选;通过序列测定后与Genebank进行同源性比较.结果 消减文库扩增获得1500余个白色阳性克隆,随机挑取95个行PCR扩增,81%的克隆中有100～600 bp的插入片段,经反Northern杂交筛选出12个差异表达基因.结论 利用立体定向技术成功制备偏侧帕金森病SD大鼠模型;利用SSH法及T/A克隆技术成功构建偏侧帕金森病大鼠纹状体组织差异表达基因消减cDNA文库.     

利用消减抑制杂交技术构建MPTP诱导的C57BL小鼠全脑差异表达基因文库

本研究旨在构建1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的C57BL小鼠全脑差异表达基因文库,并获取差异表达的表达序列标签.通过MPTP腹腔注射建立C57BL小鼠帕金森病模型.利用消减抑制杂交技术建立对照组与实验组差异表达基因文库.自两文库中各随机挑选20个克隆,经反Northern杂交证实其中7个克隆为阳性.同时对上述40个克隆进行序列测定,并与已知序列基因库进行同源性比较,其中26个新基因片段向Genbank申请了序列号.     

利用消减抑制杂交技术构建MPTP诱导的C57BL小鼠全脑差异表达基因文库

本研究旨在构建 1 甲基 4 苯基 1,2 ,3,6 四氢吡啶 (MPTP)诱导的C5 7BL小鼠全脑差异表达基因文库 ,并获取差异表达的表达序列标签。通过MPTP腹腔注射建立C5 7BL小鼠帕金森病模型。利用消减抑制杂交技术建立对照组与实验组差异表达基因文库。自两文库中各随机挑选 2 0个克隆 ,经反Northern杂交证实其中 7个克隆为阳性。同时对上述 4 0个克隆进行序列测定 ,并与已知序列基因库进行同源性比较 ,其中 2 6个新基因片段向Genbank申请了序列号。     

新生大鼠小脑内发育相关基因的筛选

为筛选脑发育相关的基因,应用新发展的抑制消减杂交技术,以新生大鼠小脑mRNA逆转录cDNA作为测试者(tester),成年大鼠小脑mRNA逆转录cDNA作为驱动者(driver),进行消减杂交,克隆制备成新生大鼠小脑高表达的cDNA文库.挑选出50个重组质粒,测序得到37个不同基因片段序列.再次用反Northern杂交确认有意义的差异表达基因.同时将测序结果与GenBank注册序列进行同源性比较,20个大鼠中首次测得的基因表达序列标签(EST)已被GenBank收录(BG695726-BG695745).     

单眼剥夺鸽视盖与人CDC10同源基因的克隆与表达分析

初步研究鸽视觉偏侧化的分子机制。建立鸽左侧单眼剥夺模型，采用抑制消减杂交和反Northern方法筛选右视盖差异表达的表达序列标签。自右视盖消减文库随机取60个重组克隆进行反Northern杂交，证实为真阳性克隆6个，随机对其一进行序列分析。发现该片段与人CDC10有84%的同源性。CDC 10属MBF转录因子复合体的一部分，介导细胞周期调节的转录，该表达序列标签在右视盖表达增强，可能调制一些视觉相关神经元的分化、增殖，其与鸽视觉偏侧化的关系需进一步克隆其全序列了解其功能方可定论。     

新生大鼠小脑内发育相关基因的筛选

为筛选脑发育相关的基因 ,应用新发展的抑制消减杂交技术 ,以新生大鼠小脑mRNA逆转录cDNA作为测试者 (tester) ,成年大鼠小脑mRNA逆转录cDNA作为驱动者 (driver) ,进行消减杂交 ,克隆制备成新生大鼠小脑高表达的cDNA文库。挑选出 5 0个重组质粒 ,测序得到 37个不同基因片段序列。再次用反Northern杂交确认有意义的差异表达基因。同时将测序结果与GenBank注册序列进行同源性比较 ,2 0个大鼠中首次测得的基因表达序列标签 (EST)已被GenBank收录 (BG6 95 72 6 -BG6 95 74 5 )。     

从癫痫模型大鼠海马神经元中克隆caspase 3的新底物PIAS1

目的从癫痫模型大鼠海马凋亡神经元中克隆caspase-3的新底物.方法制作癫痫模型大鼠海马组织cDNA文库;PCR获得caspase-3的P12和P17两亚基,然后分别定向插入pBridge质粒,构建三杂交诱饵载体;进行酵母三杂交筛库.结果建立成功癫痫模型大鼠海马组织cDNA文库,构建了大鼠caspase-3基因的酵母三杂交诱饵载体,并通过了caspase-3与eIF2α之间的相互作用验证,筛库获得caspase-3的新底物PIAS1.结论用酵母三杂交系统寻找caspase-3下游底物具有可行性,从癫痫模型大鼠海马的cDNA文库中筛库获得caspase-3的新底物PIAS1,为进一步研究caspase-3在癫痫发作引起的海马损伤中的作用奠定了基础.     

从癫痫模型大鼠海马神经元中克隆caspase 3的新底物PIASI

目的 从癫痫模型大鼠海马凋亡神经元中克隆caspase-3的新底物。方法 制作癫痫模型大鼠海马组织cDNA文库；PCR获得caspase-3的P12和P17两亚基，然后分别定向插入pBridge质粒，构建三杂交诱饵栽体；进行酵母三杂交筛库。结果 建立成功癫痫模型大鼠海马组织cDNA文库，构建了大鼠caspase-3基因的酵母三杂交诱饵栽体，并通过了caspase-3与eIF2n之间的相互作用验证，筛库获得caspase-3的新底物PIASI。结论 用酵母三杂交系统寻找caspase-3下游底物具有可行性，从癫痫模型大鼠海马的cDNA文库中筛库获得caspase-3的新底物PIASl，为进一步研究caspase-3在癫痫发作引起的海马损伤中的作用奠定了基础。     

caspase-3在颞叶癫痫大鼠海马神经凋亡中作用的实验研究

目的构建癫痫模型大鼠海马凋亡神经元的cDNA文库,采用酵母三杂交技术从中寻找caspase-3潜在的酶切底物。方法制作海人藻酸大鼠癫痫模型,取其海马组织制作酵母三杂交cDNA文库,克隆获得caspase-3的P12、P17两亚基,然后分别定向插入同一pBridge质粒,构建caspase-3的酵母三杂交诱饵载体并进行验证,然后对所构建文库进行筛库。结果成功构建癫痫模型大鼠海马组织cDNA文库,构建了caspase-3酵母三杂交诱饵载体,并通过caspase-3与eIF2α之间的相互作用验证,经筛库获得caspase-3的底物MIZ1。结论酵母三杂交技术可用于寻找caspase-3下游底物,从癫痫模型大鼠海马的cDNA文库中筛库获得caspase-3的底物MIZ1,为进一步研究caspase-3在癫痫发作引起的海马神经元损伤中的作用奠定了基础。     

The effect of age of onset of PD on risk of dementia

Background Dementia occurs in the majority of patients with Parkinson’s disease (PD). Late onset of PD has been reported to be associated with a higher risk for dementia. However, age at onset (AAO) and age at baseline assessment are often correlated. The aim of this study was to explore whether AAO of PD symptoms is a risk factor for dementia independent of the general effect of age. Methods Two community-based studies of PD in New York (n = 281) and Rogaland county, Norway (n = 227) and two population-based groups of healthy elderly from New York (n = 180) and Odense, Denmark (n = 2414) were followed prospectively for 3–4 years and assessed for dementia according to DSM-IIIR. All PD and control cases underwent neurological examination and were followed with neurological and neuropsychological assessments. We used Cox proportional hazards regression based on three different time scales to explore the effect of AAO of PD on risk of dementia, adjusting for age at baseline and other demographic and clinical variables. Findings In both PD groups and in the pooled analyses, there was a significant effect of age at baseline assessment on the time to develop dementia, but there was no effect of AAO independent of age itself. Consistent with these results, there was no increased relative effect of age on the time to develop dementia in PD cases compared with controls. Interpretation This study shows that it is the general effect of age, rather than AAO that is associated with incident dementia in subjects with PD. Received in revised form: 22 December 2005     

帕金森病运动症状进展分析

目的分析帕金森病(PD)患者运动症状进展特点。方法采用PD统一评分量表(UPDRS)Ⅲ对912例PD患者进行评估。结果与病程1年的患者比较,除病程1~2年的患者外,其他病程患者的UPDRSⅢ评分、强直分、姿势或步态异常分、轴性症状总分、言语分、步态分显著升高(均P0.05),病程5~6年及14年患者的震颤分,病程5~6年、7~8年、9~13年、14年患者的运动迟缓分、姿势分显著升高(P0.05~0.01)。轴性症状进展速度高于UPDRSⅢ评分。结论 PD患者病程早期UPDRSⅢ评分进展快,震颤症状进展独立于其他症状,轴性症状评分较UPDRSⅢ更敏感地反映疾病加重趋势。     

Frequency of accumulation of filaments in adaxonal cytoplasm of Schwann cells of myelinated fibers of rat spinal roots

Summary The frequency of accumulation of 6-nm filaments in the adaxonal cytoplasm of Schwann cells in the 6th lumbar dorsal and ventral roots was evaluated in 4-, 8-, 26- and 45-week-old Sprague-Dawley rats. The frequency was higher in 4- and 8-week-old (growing) rats than in 26- and 45-week old (mature) rats, and also higher in ventral than in dorsal roots in 4-, 8- and 26-week old rats. There were no clusters on certain groups of myelinated fibers according to the size of transverse axonal area, in both the ventral and dorsal roots. Therefore, this accumulation may reflect certain functions of the adaxonal cytoplasm of Schwann cell during natural growth and maturation of the axon and myelin sheath.     

Function of circle of Willis

Nearly 400 years ago, Thomas Willis described the arterial ring at the base of the brain (the circle of Willis, CW) and recognized it as a compensatory system in the case of arterial occlusion. This theory is still accepted. We present several arguments that via negativa should discard the compensatory theory. (1) Current theory is anthropocentric; it ignores other species and their analog structures. (2) Arterial pathologies are diseases of old age, appearing after gene propagation. (3) According to the current theory, evolution has foresight. (4) Its commonness among animals indicates that it is probably a convergent evolutionary structure. (5) It was observed that communicating arteries are too small for effective blood flow, and (6) missing or hypoplastic in the majority of the population. We infer that CW, under physiologic conditions, serves as a passive pressure dissipating system; without considerable blood flow, pressure is transferred from the high to low pressure end, the latter being another arterial component of CW. Pressure gradient exists because pulse wave and blood flow arrive into the skull through different cerebral arteries asynchronously, due to arterial tree asymmetry. Therefore, CW and its communicating arteries protect cerebral artery and blood–brain barrier from hemodynamic stress.     

Effects of prevention of afferentation of the development of the chick optic lobe

BONDY, S. C., M. E. HARRINGTON AND C. L. ANDERSON. Effects of prevention of afferentation on the developmentof the chick optic lobe. BRAIN RES. BULL. 3(5) 411–413, 1978.—The effects of unilateral extirpation of the right optic cup of the three-day incubated chick embryo upon the rate of synthesis and the stability of DNA in the non-innervated optic lobe, have been studied. This surgical procedure prevents innervation of the optic lobe contralateral to the removed eye, while the other optic lobe is normally innervated by retinal ganglion cells of the remaining eye. At the 20th day of incubation, the DNA content of the non-innervated lobe was below that of the paired lobe receiving normal innervation. This deficiency of cell number was caused by two events; death of an excess number of neurons formed early in embryogenesis and a reduced rate of glial proliferation in the later stages of incubation.     

腺垂体功能减退症临床特征变化分析

目的探讨腺垂体功能减退症患者的病因结构变化及临床表现。方法回顾性分析我院2013-01—2016-12住院及门诊78例腺垂体功能减退症患者的临床资料。结果男32例(41.03%),女46例(58.97%);诊断时年龄11~89岁,平均62.5岁;鞍区占位(包括术前及术后)52例(66.67%),席汉综合征8例(10.26%),空泡蝶鞍9例(11.65%),病因不明8例(10.26%),垂体-下丘脑发育不良1例(1.28%)。首次就诊科室:纳差厌食、恶心呕吐就诊于消化内科36例(46.15%)最常见。ACTH+TSH+Gn+G激素缺乏为19例最多,占24.36%,ACTH+TSH+Gn缺乏15例,占19.23%。结论腺垂体功能减退症病因结构发生变化,发病人群、首发症状及受累激素也不同,患者女性多于男性,发病年龄偏高,症状不典型,分布于临床多个科室,其中以低钠血症为首发临床表现就诊消化内科最多。     

Standards of instrumentation of EMG

Standardization of Electromyography (EMG) instrumentation is of particular importance to ensure high quality recordings. This consensus report on “Standards of Instrumentation of EMG” is an update and extension of the earlier IFCN Guidelines published in 1999. First, a panel of experts in different fields from different geographical distributions was invited to submit a section on their particular interest and expertise. Then, the merged document was circulated for comments and edits until a consensus emerged.The first sections in this document cover technical aspects such as instrumentation, EMG hardware and software including amplifiers and filters, digital signal analysis and instrumentation settings. Other sections cover the topics such as temporary storage, trigger and delay line, averaging, electrode types, stimulation techniques for optimal and standardised EMG examinations, and the artefacts electromyographers may face and safety rules they should follow. Finally, storage of data and databases, report generators and external communication are summarized.     

Modelling of movement of the lamprey: Control of oscillators

This article discusses the control methods of the central pattern generator (CPG). First a control model of the CPG is presented using 2 oscillators, and we suggest that phasic modulation to the CPG by means of phasic information is effective for controlling the phase difference between oscillators. Next, two models for controlling the CPG of a lamprey are proposed. One model describes a control system from the brain stem, in which the reticulospinal neurons control the CPG by receiving feedback signals and sending control signals to the neck region of the CPG. The other is a model for learning an localized control system to generate a desired motor pattern. By means of these models, a role of the efference copy is suggested.     

《绵阳市社会心理服务工作管理办法》解读

2018年,国家卫生健康委员会等10部委联合发布《关于印发全国社会心理服务体系建设试点工作方案的通知》,四川省绵阳市被列为全国第一批试点地区。绵阳市人民政府依据《中华人民共和国精神卫生法》等相关法律法规和文件精神,结合前期调查研究和社会心理服务工作的试点实际,编制出台了《绵阳市社会心理服务工作管理办法》,并于2021年12月25日起施行。本文围绕社会心理服务的相关概念、办法总则、重点内容、保障措施等方面进行解读,以期为社会心理服务工作的规范、持续和有效开展提供参考。     

利培酮对精神分裂症患者生活质量的影响

目的：比较利培酮与氟哌啶醇对精神分裂症患者生活质量的影响。方法：对门诊72例服用氟哌啶醇及74例服用利培酮的精神分裂症患者用生活质量综合评定问卷（GQOLI）、阳性与阴性症状量表（PANSS）、副反应量表（TESS）进行评定。结果：利培酮组患者治疗后生活质量有所提高,而氟哌啶醇组患者生活质量有所下降。结论：利培酮治疗有利于患者提高生活质量。