兔骨髓基质细胞核移植的初步研究

目的 通过建立骨髓基质细胞核移植的方法，为进一步从克隆囊胚的内细胞团细胞分离培养出全能性的胚胎干细胞并用于治疗性克隆等研究奠定基础。方法 超排获得的兔成熟卵母细胞去核后，将同种的单个骨髓基质细胞核注入其内；电融合后，经离子霉素（ionomycin）和6-甲氨基嘌呤（6-DMAP）激活，用10％胎牛血清（FBS）的TCM-199进行体外培养直至囊胚阶段。结果 实验对147个卵母细胞去核，去核成功率为70．75％（104／147）；注核后有82．69％（86／104）的卵细胞保持形态完整；重构后的体细胞-卵母细胞质复合体经电融合后的融合率为56．79％（46／81）；激活后的重构胚于体外培养后分裂率为65．22％（30／46），囊胚率为17．39％（8／46），获得的囊胚有62．5％（5／8）进入孵化阶段；孵化出来的细胞贴壁生长并向周围扩展，显示了具有向下一阶段发育的潜能，为从囊胚内细胞团细胞中分离并培养出胚胎干细胞提供了可能。结论 实验结果表明，以兔成体体细胞的骨髓基质细胞为核供体，经克隆技术生产出囊胚并从中分离内细胞团细胞具有可行性。     

人骨髓基质细胞作为核移植供体细胞的相关生物学特性研究

目的研究作为核移植供体细胞的人骨髓基质细胞相关生物学特性。方法应用密度梯度离心法分离培养人骨髓基质细胞，并对其进行形态观察、免疫荧光分析细胞骨架结构、端粒酶活性测定、核型及细胞周期分析。结果培养的细胞具有正常的大小、形态、细胞骨架结构：在融合密度达80%-90%时，G0＋G1期细胞所占的比例较高，约占92．58％；检测传至第7代的人骨髓基质细胞具有正常染色体数目（46，XY），端粒酶活性为0．567。结论形态良好、染色体数目正常的人骨髓基质细胞可以作为核移植的供体细胞。     

Scriptaid对小鼠体细胞核移植克隆胚早期发育能力的提高作用

目的 探讨抗肿瘤药物Scriptaid对小鼠体细胞核移植(SCNT)克隆胚体外发育的影响及提高移植效率的新途径. 方法 以C57/BL6小鼠卵母细胞为受体、卵丘细胞为供体进行SCNT,构建克隆胚,并采用数字随机表法将克隆胚分为5组,分别在含0 nmol/L(阴性对照组)、50、100、250、500 nmol/L Scriptaid的无钙激活剂中激活6h,然后在含相应浓度Scriptaid的KSOM培养基中处理4h,最后转移至KSOM培养基中孵育 96h.观察、记录各组克隆胚的发育情况并进行囊胚细胞计数. 结果 各组克隆胚激活率和2-cell率之间差异无统计学意义(P＞0.05);250 nmol/LScriptaid组的囊胚形成率(24.2％)和囊胚细胞数(56.27±2.43)与0 nmol/L组、50 nmol/L组、100nmol/L、500 nmol/L Scriptaid组之间(其囊胚形成率分别为5.3％、6.5％、9.4％和6.9％,囊胚细胞数分别为44.67±1.53、50.25±1.26、52.33±2.50和50.75±1.50)比较差异均有统计学意义(P＜0.05). 结论 250 nmol/L Scriptaid能有效提高小鼠SCNT克隆胚体外早期发育能力.     

核转染增强型绿荧光蛋白对兔原代骨髓基质细胞向神经细胞方向分化的影响

目的研究以最近发展起来的核转染技术转染增强型绿荧光蛋白（EGFP）质粒进行基因修饰对兔原代骨髓基质细胞向神经细胞方向分化的影响。方法从兔股骨抽取骨髓，密度梯度离心法获取原代骨髓基质细胞。以Nucleofector^TM技术转染pEGFP—C2（EGFP组），以同期培养未转染的细胞作为对照组（control组）。以“CYTOKINE·神经干细胞培养基”＋10％胎牛血清诱导BMSCs向神经细胞方向分化。流式细胞仪检测转染效率。比较两组细胞的生长增殖以及Nestin、NSE、GFAP抗原的表达情况。结果在转染后24h成功发现EGFP的表达。两组细胞具有相似的形态学变化以及生长曲线。诱导18d，两组细胞NSE、GFAP的表达比例无统计学意义。结论Nucleofector^TM技术是一种简易而高效的转染兔原代骨髓基质细胞的方法；EGFP可以作为兔骨髓基质细胞有效的基因表达标记；核转染pEGFP-C2对兔原代骨髓基质细胞体外增殖及向神经细胞方向分化无明显影响。     

经枕下-乙状窦后入路显微手术治疗大型听神经瘤

目的探讨经枕下-乙状窦后入路显微手术切除大型听神经瘤的操作技术及预后。方法对26例大型听神经瘤行显微手术切除，术中及术后评估面神经、听神经的功能。结果本组大型听神经瘤26例，其中21例（80．8％）全切除，5例（19．2％）次全切除。面神经的解剖学保留率为73．1％（19／26）。对23例进行1年以上的随访评估，面神经功能获得满意恢复者为18例（78．3％）。结论采用显微手术切除大型听神经瘤，对面神经、听神经可获得解剖学与功能的保护，明显降低并发症与死亡率，收到满意的临床疗效。     

局部激素缓释片应用对脑内血肿清除后脑水肿的影响

目的研究局部应用地塞米松缓释片对兔脑内血肿清除术后脑水肿的影响。方法32只兔子随机分为A（对照组）和B（治疗组）两组并应用自体血注射法建立脑内血肿模型。A组行常规血肿清除术；B组于血肿清除术后，在血肿残腔内置入地塞米松缓释片。检测两组兔脑组织含水量和血清髓鞘碱性蛋白（MBP）水平并比较。结果A、B两组患侧大脑半球含水量分别为（83．61±0．62）％和（81．53±0．59）％，B组显著低于A组（P〈0．05）。A、B两组血清MBP分别为（6．18±2．13）μg／L和（3．35±1．29）μg／L，均高于正常对照值（1．78±0．76）μg／L，且A组显著高于B组（P〈0．05）。结论脑内血肿清除术后脑组织含水量及血清MBP增高，局部应用地塞米松缓释片后显著降低，提示该方法是预防和治疗脑内血肿清除术后脑水肿的有效方法。     

内镜辅助的眶上锁孔人路显微手术切除鞍区病变（附52例报告）

目的总结内镜辅助的眶上锁孔入路显微手术切除鞍区病变的经验、手术方法与技巧。方法对52例鞍区病变均采用内镜辅助的眶上锁孔入路显微手术切除。此52例鞍区病变中，垂体腺瘤22例，颅咽管瘤8例，脑膜瘤7例及前循环不同类型动脉瘤15例。结果37例鞍区肿瘤中，肿瘤全切除31例（83．8％，31／37），次全切除或部分切除6例（16．2％，6／37）。15例动脉瘤均准确完全夹闭瘤颈，预后优良者13例（86．7％13／15）和轻残2例（1313％，2／15）。全组病人无手术死亡及重残，且术前原有症状在术后均有明显不同程度的好转。结论内镜辅助的眶上锁孔入路显微手术能清楚显露鞍区病变与周围结构，创伤小，避免了重要结构的损伤，术后并发症少，效果好。     

胎鼠皮质神经元细胞核的制备和鉴定

目的 建立制备皮质神经元细胞核的方法，以便用于核移植研究。方法 （1）采用胎龄17d的胎鼠进行皮质神经元的分离培养，用抗MAP2和NeuN免疫细胞化学法进行鉴定；（2）用梯度离心法制备神经元细胞核，并用HE染色、免疫荧光染色、透射电镜检查等方法进行鉴定，用台盼蓝染色检测其活性。结果 （1）胎鼠皮质神经元呈多角形，伸出多个突起，MAP2染色显示神经元细胞浆和突起呈免疫反应阳性，NeuN染色显示神经元细胞核呈免疫反应阳性，部分核周胞质和部分近端突起也呈免疫反应阳性，MAP2和NeuN阳性细胞比例均在90％左右；（2）HE染色和Hoechst33342免疫荧光染色显示神经元梯度离心后在22％～30％Ficoll400梯度层可见到大量圆点状细胞核，台盼蓝染色显示活细胞核比例为99．6％；在22％～30％Ficoll400梯度层细胞透射电镜标本中可观察到圆形的细胞核，在细胞核的周围仅见到一层薄薄的胞浆成分。结论 梯度离心法是制备皮质神经元细胞核的可靠方法。     

大型桥小脑角脑膜瘤显微手术治疗及神经功能保护

目的探讨大型（直径〉4cm）桥小脑角脑膜瘤显微外科手术切除方法以及神经功能保护效果。方法对47例经显微外科手术治疗的大型桥小脑角脑膜瘤患者的临床资料进行回顾性分析。结果47例中，行SimpsonⅠ、Ⅱ级切除40例（85．1％），部分切除7例，无手术死亡病例。面神经解剖保留率为89．4％（42／47），面神经功能（House—BmekmannⅠ、Ⅱ级）保留率为76．6％（36／47），听神经解剖保留率为83．0％（39／47），听力保留率66．0％（31／47）．结论采用枕下乙状窦后人路显微手术切除大型桥小脑角脑膜瘤安全、有效，在保留神经功能完整性的前提下应尽量切除肿瘤。     

术前影像学在垂体泌乳素腺瘤治疗中的意义

目的垂体泌乳素腺瘤临床治疗策略各家观点不一。本文旨在分析总结影像学在该疾病治疗策略确定中的潜在作用。方法对5年间326例垂体泌乳素腺瘤病例的临床表现、影像学特点、治疗策略的确定及疗效预后进行回顾性分析。结果平均年龄男性41．6岁，女性32．3岁。男性肿瘤直径平均38．05mm，其中83．3％肿瘤呈侵袭性生长，在女性则为15．25mm和49％。手术组平均血清泌乳素（PRL）水平男性1875．22ng／ml，女性376．6ng／ml。术后，男性降至385．84ng／ml（P〈0．05），女性PRL降至23．93ng／ml（P〈0．05）。微腺瘤不孕者术后受孕率达75％，高于药物治疗组33．33％（P〈0．05）。结论对于影像学上存在明显病灶的病例，显微手术可取得良好疗效，若术后血清激素控制不良或海绵窦残留侵袭肿瘤的病例可考虑辅助药物或放射治疗。对于影像学明确提示微腺瘤，尤其是有生育要求的女性病例，积极的显微手术可取得较好疗效。     

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。