TRAIL对原代培养胶质瘤细胞的凋亡诱导作用

目的研究TRAIL对原代培养的胶质瘤细胞的凋亡诱导作用，探讨其临床应用的实际价值。方法对22例脑胶质瘤和5例正常脑组织进行原代培养，用流式细胞仪检测TRAIL作用后肿瘤细胞和正常肢质细胞的凋亡率。结果成功培养出18例胶质瘤细胞和4例正常胶质细胞。800ng／ml,TRAIL作用48h后，凋亡率在80％以上的5例，50％。80％ 9例。50％以上14例．占总数的77．78％，50％以下的4例；正常胶质细胞无明显凋亡发生。结论TRAIL能有效地选择性地杀死某些胶质瘤细胞，对正常胶质细胞则无损害作用，TRAIL为肢质瘤的安全治疗提供了一个新的备选方法。     

顺铂在TRAIL/APO-2L致胶质瘤细胞凋亡作用中影响机制的研究

目的探讨TRAIL对胶质瘤细胞凋亡的影响,并进一步研究DNA破坏药物顺铂(CDDP)在与其联合作用诱导细胞凋亡中影响作用的机制。方法行人脑胶质瘤U251细胞株的培养,应用不同浓度TRAIL和CDDP作用于胶质瘤U251细胞,MTT法检测TRAIL和CDDP分别及联合作用于U251细胞后的存活率,并以亚毒剂量的TRAIL和CDDP联合作用后,流式细胞仪检测其凋亡率。Westernblot方法检测CDDP作用前后TRAIL受体DR5表达量的变化。结果MTT结果:在一定时间范围内,随着药物浓度增加,TRAIL和CDDP分别及联合作用皆使胶质瘤U251细胞存活率逐渐降低,其中两者的联合作用效果明显,与分别作用组差异有统计学意义(P<0.01)。亚毒剂量的TRAIL(50ng·ml-1)、CDDP(4μg·ml-1)分别及联合作用人脑胶质瘤U251细胞24h后,存活率分别为:72.43%,75.56%,23.45%。亚毒剂量的TRAIL(50ng·ml-1)、CDDP(4μg·ml-1)单独或联合作用组致胶质瘤细胞凋亡率分别为25.61%、20.59%、62.33%。单独应用组与联合组之间差异有统计学意义(P<0.01)。Westernblot法检测显示:CDDP作用后较作用前DR5表达有明显增高,随CDDP剂量增多,DR5表达量逐渐增加。结论TRAIL和CDDP联合作用能显著增强其对人U251胶质瘤细胞凋亡的诱导,其机制可能与CDDP能上调TRAIL死亡受体DR5的表达有关。     

高浓度谷氨酸诱导胶质瘤细胞凋亡及其机制

目的：研究高浓度谷氨酸对原代培养人脑胶质瘤细胞的凋亡诱导作用,进一步探讨其凋亡的可能机制。方法：对新鲜的人脑胶质瘤进行体外原代培养,不同浓度的谷氨酸作用不同时间,形态学观察;四甲基偶氮唑盐（MTT）法检测肿瘤细胞生长抑制率;流式细胞术检测肿瘤细胞凋亡率;激光共聚焦显微镜检测细胞膜内游离钙离子的浓度变化。结果：原代培养胶质瘤细胞的生长抑制率随谷氨酸浓度的增加呈递增趋势。25、50和100mmol/L谷氨酸作用胶质瘤细胞48h的凋亡率分别为9.65%、31.13%和20.35%。脑胶质瘤原代培养细胞内钙离子浓度在谷氨酸作用后明显升高,与对照组相比有显著性差异（P〈0.01）;但不同浓度组、24h和48h组之间钙离子的浓度无显著性差异（P〉0.05）。结论：高浓度的谷氨酸可诱导原代培养人脑胶质瘤细胞凋亡,凋亡率在一定范围内呈明显的剂量依赖性。钙离子在谷氨酸诱导的细胞凋亡中起重要作用。     

三氧化二砷诱导C6胶质瘤细胞凋亡实验研究

目的：研究三氧化二砷（As2O3）在体外是否具有抗鼠胶质瘤作用,并对其作用机制进行初步探讨。方法：应用不同浓度As2O3,且在不同时间点分别处理C6胶质瘤细胞株及原代培养的正常鼠神经胶质细胞,采用甲基噻唑基四唑（MTT）法,观察As2O3对C6胶质瘤细胞株及正常鼠神经胶质细胞生长的影响,以透射电镜、Hoechst33342和碘化丙啶（PI）双重荧光染色检测两种细胞凋亡的形态变化,并用异硫氰酸荧光素（FITC）标记的Annexin-V和PI双标记法通过流式细胞仪检测细胞凋亡率。结果：MTT法发现,As2O30.5～8.0μmol/L的浓度均可显著抑制C6胶质瘤细胞株的生长,而对正常原代神经胶质细胞株的抑制作用较弱。经As2O3作用后,透射电镜、Hoechst33342/PI双染荧光均观察到C6胶质瘤细胞发生了显著的凋亡形态改变;运用Annexin-V-FITC/PI双标记法在流式细胞仪检测显示,随As2O3浓度的增大和时间的延长,C6胶质瘤细胞株的凋亡率明显上升,而正常神经胶质细胞的凋亡率要明显小于C6胶质瘤细胞株。结论：As2O3在体外可显著抑制C6胶质瘤细胞株生长,其机制与诱导肿瘤细胞凋亡有关,且凋亡率随As2O3作用的时间和剂量的增加而增加。但As2O3对正常神经胶质细胞生长的抑制作用较弱,提示As2O3抑制细胞生长的作用具有一定的选择性。     

TRAIL诱发人脑H4神经胶质瘤细胞凋亡的研究

目的研究TRAIL对H4神经胶质瘤细胞的凋亡诱导作用，并探讨其可能机制。方法利用显微镜、透射电镜、流式细胞仪，观察和检测TRAIL对传代培养的肿瘤细胞的凋亡诱导作用，用MTT法检测easpase-8抑制剂zIETD—fmk对TRAIL凋亡诱导作用的抑制情况。结果TRAIL作用后镜下可见到H4细胞凋亡的典型表现；在流式细胞仪检测PI荧光直方图上，出现典型的亚二倍体峰，随着作用时间的延长，H4细胞凋亡率明显增加；zIETD—fmk能显著抑制TRAIL对H4细胞的凋亡诱导作用。结论TRAIL可有效的诱导H4神经胶质瘤细胞凋亡；easpase-8在H4瘤细胞的凋亡过程发挥着重要作用，TRAIL可能是通过激活caspase系统而诱发H4瘤细胞凋亡的。     

膜结合型肿瘤坏死因子相关凋亡诱导配体修饰的人骨髓基质干细胞对脑胶质瘤细胞的体外靶向抗肿瘤作用

目的：探索经膜结合型肿瘤坏死因子相关凋亡诱导配体（TRAIL）修饰的人骨髓基质干细胞（hMSCs）对脑胶质瘤的靶向抗肿瘤作用。方法：通过体外Transwell系统研究hMSCs向胶质瘤的迁徙能力。采用经全长人TRAIL（hTRAIL）重组的腺相关病毒（rAAV）载体（rAAV hTRAIL）对骨髓基质干细胞（MSCs）进行修饰。将hTRAIL修饰的MSCs与人胶质瘤细胞（U87MG）在体外共同培养,分析其对脑胶质瘤的靶向抗肿瘤作用。结果：hMSCs向胶质瘤细胞的迁徙具有高度特异性。经TRAIL修饰的hMSCs不仅在MSCs表面表达全长TRAIL,且能在培养基中分泌可溶性TRAIL。经rAAV-hTRAIL转染后,hMSCs细胞凋亡作用无增加。将TRAIL修饰的hMSCs与U87MG细胞共培养,与采用可溶性TRAIL治疗后相比较,可明显提高U87MG细胞的凋亡率。结论：在体外实验中,hMSCs向胶质瘤细胞的迁徒具有高度特异性,且能诱导胶质瘤细胞凋亡,并提高对sTRAIL不敏感胶质瘤细胞的凋亡率。由此推测膜结合型TRAIL修饰的MSCs在肿瘤微环境中对脑胶质瘤具备靶向抗肿瘤效应。MSCs可能成为脑胶质瘤靶向治疗的有效载体。     

溶血磷脂酸诱导原代培养的小鼠小胶质细胞的凋亡

目的 观察不同浓度的溶血磷脂酸对原代培养小鼠小胶质细胞凋亡的干预与调控作用。方法 分离新生小鼠的大脑小胶质细胞进行原代培养，用CD11b抗原抗体免疫组化染色法鉴定纯度，在不同浓度溶血磷脂酸作用下以MTT法检测细胞增殖率，流式细胞术检测凋亡率。结果 原代培养的小胶质细胞纯度大于95％，低浓度溶血磷脂酸（0．001～20μmol／L）对小胶质细胞有激活增殖的作用；在浓度为20-80μmol／L之间时随着溶血磷脂酸浓度的增加，凋亡率逐渐升高；与对照组比较均具有显著性差异（P〈0．05）。结论 低浓度的LPA可激活小胶质细胞，促使其增殖，而高浓度的溶血磷脂酸可诱导小胶质细胞凋亡。     

小干扰RNA沉默RHBDF1基因抑制胶质瘤细胞生长和诱导细胞凋亡的初步研究

目的 检测RHBDF1基因在正常胶质细胞和胶质瘤细胞内的表达,以及小干扰RNA沉默C6胶质瘤细胞内RHBDF1基因后是否诱导C6细胞凋亡和抑制细胞生长,为基因治疗胶质瘤寻找新的靶基因. 方法采用Western blot检测RHBDF1基因在正常胶质细胞和胶质瘤细胞(包括C6、U251和MGR2)的表达情况:用脂质体转染法分别将RHBDF1 siRNA或对照siRNA转染入C6细胞内,RT-PCR和Western blot方法检测siRNA转染后对C6细胞内RHBDF1基因和蛋白的影响;Tune1法检测RHBDF1基因沉默后对细胞凋亡的诱导情况,Ki-67免疫荧光染色观察RHBDF1基因沉默后对细胞生长抑制的影响.结果 与正常胶质细胞比较,胶质瘤细胞内RHBDF1基因存在着过度表达.siRNA转染入C6细胞后.对照siRNA组凋亡率为2.96%±1.25%,而RHBDF1siRNA1组和RHBDF1 siRNA2的凋亡率分别为36.35%±4.85%和33.58%±4.08%:对照siRNA组细胞增殖率为95.96%±3.25%,而RHBDF1 siRNA1组和RHBDF1 siRNA2组的细胞增殖率分别为24.57%±5.53%和25.68%±4.08%;组间细胞凋亡率和增殖率比较差异均有统计学意义(P<0.05).结论 RHBDF1基因在胶质瘤细胞内存在过度表达,沉默该基因后可以诱导细胞凋亡和抑制细胞生长,RHBDF1基因可能成为基因治疗胶质瘤的一个新的靶基因.     

TRAIL联合美伐他汀增强对SWO-38细胞的抑瘤作用及其机制研究

目的 探讨肿瘤坏死因子相关细胞凋亡诱导配体(TRAIL)单独或联合美伐他汀对人胶质瘤细胞SWO-38增殖和凋亡的影响及其作用机制. 方法 人胶质瘤细胞SWO-38分别用人重组可溶性TRAIL蛋白(rsTRAIL)、美伐他汀及两者联用处理;MTT法检测细胞增殖情况;双染色流式细胞仪检测细胞凋亡情况;间接免疫荧光染色结合流式细胞技术检测细胞表面TRAIL R_1/DR_4和TRAIL R_2/DR_5分子表达;RT-PCR方法检测TRAIL R_1/DR_4和TRAIL R_2/DR_5 mRNA表达. 结果 rsTRAⅡ,和美伐他汀在单用或联用时均可抑制SWO-38细胞增殖,诱导其凋亡,并在一定范围内呈时间、剂量依赖性;联合用药组细胞增殖抑制率和凋亡率明显高于单用rsTRAIL或美伐他汀组,比较差异有统计学意义(P<0.05).美伐他汀单独或联合rsTRAIL作用于SWO-38细胞72 h后,死亡受体TRAIL R_1/DR_4和TRAIL R_2/DR_5在细胞表面的表达明显较对照组细胞增强,其mRNA表达水平亦相应增加,比较差异均有统计学意义(P<0.05). 结论 rsTRAIL与美伐他汀联合使用可增强对人胶质瘤细胞SWO-38的增殖抑制和凋亡诱导效应,其机制可能是美伐他汀增加SWO-38细胞TRAIL R_1/DR_4和TRAIL R_2/DR_5 mRNA表达、上调细胞表面TRAIL死亡受体从而增加其对rsTRAIL的敏感性.     

肿瘤坏死因子相关凋亡诱导配体联合化疗药物对胶质母细胞瘤细胞的杀伤作用

目的研究肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor related apoptosis-inducing ligand,TRAIL)及联合化疗药物对胶质母细胞瘤细胞的杀伤作用。方法培养GC427、GC125细胞,应用TRAIL及化疗药物顺铂及放线菌素D分别及联合作用于细胞后,ELISA方法检测细胞凋亡水平。结果 GC427、GC125细胞经TRAIL、顺铂、放线菌素D作用后,GC427对TRAIL抵抗,GC125对TRAIL稍敏感。顺铂与放线菌素D单独作用细胞,均可诱导两株细胞发生部分凋亡;TRAIL与顺铂或放线菌素D联合作用细胞,则可诱导细胞发生明显的凋亡反应,结果有统计学差异(P<0.05)。结论胶质母细胞瘤细胞经TRAIL与化疗药物联合作用后可发生协同效应。     

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     

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.     

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.     

Release of endogenous catecholamines from slices of hypothalamus of rats

The release of endogenous catecholamines from superfused slices of rat hypothalamus was studied under basal conditions and during release evoked by 40 mM K⁺. Catecholamines in superfusates, and in extracts of the tissue after stimulation, were isolated by column chromatography and quantitated by liquid chromatography with electrochemical detection. Norepinephrine (NE) was not consistently demonstrable in superfusate collected under basal conditions, but 40 mM K⁺ caused the release of from 2 to 4 ng/g of tissue per min. The addition of cocaine to the superfusate caused increases in basal and evoked release of NE. Epinephrine (E) could be measured in superfusates of slices from male but not female rats and then only when cocaine was added to the superfusate. Accordingly, the concentration of E in hypothalamus was greater in male rats than in female rats. Dopamine (DA) was not consistently measurable in the spontaneous overflow from slices either in the presence or absence of cocaine. K⁺-evoked release of DA could be demonstrated in slices from female rats. The addition of cocaine increased the evoked release of DA from slices from both sexes. Corticosterone, added to cocaine, had no effects on the efflux of any of the catecholamines. The experiments suggest that neuronal reuptake of all catecholamines is very efficient in the hypothalamus both under basal conditions and during evoked release.     

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.     

《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。     

阿立哌唑对精神分裂症患者生活质量的影响

目的:比较阿立哌唑与利培酮对精神分裂症患者生活质量的影响。方法:60例精神分裂患者随机平分为两组各30例,分别给予阿立哌唑和利培酮治疗。疗程8周。用生活质量综合评定问卷-74(GQOLI-74)、阳性与阴性症状量表(PANSS)及副反应量表(TESS)评定疗效及不良反应。结果:阿立哌唑与利培酮均能显著提高精神分裂症患者生活质量,但阿立哌唑在改善GQOLI-74总分、躯体健康及社会功能维度优于利培酮。结论:阿立哌唑治疗有利于提高精神分裂症患者生活质量。