氧化损伤星形胶质细胞与骨髓基质细胞共培养的实验研究

目的 观察氧化损伤星形胶质细胞与骨髓基质细胞(BMSCs)共培养时细胞损伤恢复及存活能力的改变,以进一步探讨BMSCs修复脑损伤的机制.方法 过氧化氢(H2O2)损伤体外培养的星形胶质细胞,与BMSCs共培养后光镜下观察细胞生存情况.分别在培养后第1、3天取细胞培养液应用EuSA试剂盒测定碱性成纤维细胞生长因子(bFGF)、脑源性神经营养因子(BDNF)、神经生长因子(NGF)以及表皮生长因子(EGF)的分泌水平,并与氧化损伤星形胶质细胞单独培养组和BMSCs单独培养组进行比较.结果 培养后3 d和7 d,共培养组星形胶质细胞的恢复及两种细胞的存活情况明显优于单独培养组.培养后1 d和3 d,共培养组细胞培养液中NGF、BDNF以及bFGF的浓度明显高于两个单独培养组之和,但是共培养组培养液中EGF的浓度明显低于两个单独培养组之和.结论 氧化损伤星形胶质细胞和BMSCs共培养能够提高星形胶质细胞的存活能力,增加NGF、BDNF以及bFGF的分泌水平.BMSCs移植修复脑损伤的功能可能部分是通过促进损伤星形胶质细胞恢复、减少星形胶质细胞死亡、增加神经营养和生长因子分泌完成.     

骨髓基质细胞体外分离培养和分化为

摘要 目的：观察体外培养大鼠骨髓基质细胞（BMSCs bone marrow stromal cells）的生物学特性并探讨其分化为Schwann细胞表达S100的机制,为组织工程学修复周围神经损伤奠定一定的实验基础。方法：从成年SD大鼠的股骨和胫骨中分离培养BMSCs,倒置显微镜下动态观察细胞的生长状态,采用MTT法检测细胞的活力,FCM检测细胞周期等方法研究BMSCs的生物学特性。应用复合诱导因子（BME, RA ,FSK, bFGF, PDGF, HRG）体外诱导BMSCs向Schwann细胞分化。诱导分化后采用免疫荧光细胞化学染色,流式细胞仪,逆转录PCR方法分别检测检测胶质细胞标记蛋白S100蛋白和mRNA的表达。结果：BMSCs在体外容易扩增,传1－8代以内的细胞增殖能力无明显变化。未经诱导的BMSCs大部分处于G0和G1期。诱导分化后的细胞形态上类似Schwann细胞,并且表达胶质细胞的标记蛋白S100蛋白及其mRNA明显升高。结论：BMSCs在体外可以向类Schwann细胞诱导分化,并且表达胶质细胞的标记蛋白S-100蛋白及其mRNA。     

慢病毒介导的胶质细胞系源性神经营养因子在骨髓基质细胞中的表达及其对乳胞素干预PC12细胞的保护作用(英文)

目的建立慢病毒介导的胶质细胞系源性神经营养因子(glial cell line-derived neurotrophic factor,GDNF)表达系统,体外感染骨髓基质细胞,检测过表达GDNF对蛋白酶抑制剂引起的PC12细胞损伤的神经保护作用。方法经双酶切和T4连接酶构建pLenti6/V5-GDNF表达质粒,经293FT细胞包装产生高滴度病毒。用RT-PCR、ELISA和免疫细胞化学方法检测感染骨髓基质细胞(bone marrow stromal cells, BMSCs)后GDNF的表达,并检测过表达GDNF对蛋白酶抑制剂乳胞素(lactacystin)引起的PC12细胞损伤的保护作用。结果成功构建pLenti6/V5-GDNF表达质粒,获得高滴度具有感染能力的病毒储存液(5.6×105TU/mL)。BMSCs体外被感染后能大量分泌GDNF(接近800pg/mL),过表达GDNF能减轻乳胞素(10μmol/L)引起的PC12细胞损伤。结论慢病毒介导的GDNF转染骨髓基质细胞后能分泌具有生物学活性的GDNF,对蛋白酶体抑制剂引起的PC12细胞损伤有保护作用。     

氟西汀对大鼠星形胶质细胞分泌的胶质源性神经营养因子的影响

目的 探讨氟西汀对大鼠星形胶质细胞分泌的胶质源性神经营养因子(GDNF)的影响.方法 以氟西汀干预体外培养的大鼠海马星形胶质细胞,通过四甲基偶氮唑盐法(MTT)检测不同浓度氟西汀对细胞活力的影响;采用酶联免疫吸附测定法(ELISA)检测细胞培养液GDNF浓度及Real-time PCR法检测GDNFmRNA的表达.结果 (1)氟西汀浓度超过35 μmol/L浓度时,可降低细胞活性,差异有统计学意义(P<0.01或P<0.05);(2)10 μmol/L氟西汀干预星形胶质细胞不同时间后,48 h组细胞培养液GDNF浓度[(68±13)fg/L]高于0 h组[(32±11)fg/L]、6 h组[(34±12)fg/L]、12 h组[(41±17)fg/L]、24 h组[(45±13)fg/L],差异均有统计学意义(P均<0.01);(3)不同浓度氟西汀作用星形胶质细胞48 h后,10 μmol/L浓度组的细胞培养液GDNF浓度[(64±17)fg/L]高于0 μmol/L[(39±15)fg/L]和1 μmoVL浓度组[(39±18)fg/L],差异均有统计学意义(P均<0.05);(4)氟西汀作用星形胶质细胞48 h后,撤离氟西汀24 h后星形胶质细胞仍明显分泌GDNF,差异有统计学意义(P<0.01或P<0.05);(5)不同浓度氟西汀作用星形胶质细胞24 h后,10 μmol/L和20 μmol/L浓度组细胞GDNFmRNA表达量[分别为(0.008 1±0.001 1)和(0.006 3±0.000 3)]高于0 μmol/L、1 μmol/L及5 μmol/L浓度组[分别为(0.003 1±0.000 7)、(0.003 9±0.000 3)和(0.004 1±0.000 2)],差异均有统计学意义(P均<0.01).结论 氟西汀可能通过促进星形胶质细胞GDNF的分泌来发挥其神经保护作用.     

蛋白酶体抑制对培养星形胶质细胞IL-6分泌的影响

目的 研究蛋白酶体抑制对体外培养的星形胶质细胞(AS)IL-6分泌的影响.方法 SD乳鼠皮层AS原代培养,并纯化鉴定;不同浓度(0.1,1,2.5,5 μM)的蛋白酶体抑制剂(lactacystin)对第二代AS进行短期(24 h)急性处理,另一批同期的AS应用较低浓度(0.5μM)的lactacystin长期(4周)慢性处理,应用RT-PCR及ELISA方法检测AS IL-6 mRNA的表达和培养基中IL-6蛋白的分泌水平.结果 纯化传代的皮层AS经GFAP免疫荧光鉴定,其阳性率可达99.45%;短期抑制组中,1,2.5,5μM蛋白酶体抑制剂可诱导AS IL-6 mRNA表达和蛋白分泌增加,与对照组比较差异有显著性(P<0.01);长期抑制组中IL-6 mRNA表达及蛋白分泌也较同期对照组增加,差异有统计学意义.结论 一定程度蛋白酶体活性抑制可诱导培养的AS IL-6 mRNA表达及蛋白分泌增加,提示蛋白酶体功能障碍后可能通过诱导AS分泌IL-6增加来参与阿尔茨海默病的病理改变.     

慢病毒介导的胶质细胞系源性神经营养因子在骨髓基质细胞中的表达及其对乳胞素干预PC12细胞的保护作用

目的 建立慢病毒介导的胶质细胞系源性神经营养因子(glial cellline-derived neurotrophic factor,GDNF)表达系统,体外感染骨髓基质细胞,检测过表达GDNF对蛋白酶抑制剂引起的PC12细胞损伤的神经保护作用.方法 经双酶切和T4连接酶构建pLenti6/V5-GDNF表达质粒,经293FT细胞包装产生高滴度病毒.用RT-PCR、ELISA和免疫细胞化学方法检测感染骨髓基质细胞(bone marrow stromal cells,BMSCs)后GDNF的表达,并检测过表达GDNF对蛋白酶抑制剂乳胞素(1actacystin)引起的PC12细胞损伤的保护作用.结果 成功构建pLenti6/V5-GDNF表达质粒,获得高滴度具有感染能力的病毒储存液(5.6×105 TU/mL).BMSCs体外被感染后能大量分泌GDNF(接近800 pg/mL),过表达GDNF能减轻乳胞素(10 μmol/L)引起的PC12细胞损伤.结论 慢病毒介导的GDNF转染骨髓基质细胞后能分泌具有生物学活性的GDNF,对蛋白酶体抑制剂引起的PC12细胞损伤有保护作用.     

骨髓源性神经干细胞诱导分化为多巴胺能神经元的实验研究

目的探索骨髓基质细胞(BMSCs)诱导分化为神经干细胞及多巴胺能神经元的分化条件和发生机制,比较不同血清浓度、不同白介素-1(IL-1α)及不同胶质细胞源神经营养因子(GDNF)浓度及不同组合浓度等诱导条件下BMSCs分化情况;为BMSCs在神经科学领域内的应用奠定基础。方法以成年SD大鼠BMSCs为实验对象,利用IL-1α、胶质细胞系来源GDNF等作为增殖及分化诱导因子,进行增殖培养、分化诱导;免疫细胞法进行细胞性质鉴定。结果加入GDNF、IL-1 α增殖培养诱导6 d,部分细胞有神经巢蛋白成分表达;三周测出 DA受体D2检测阳性,GDNF IL-1α组与 GDNF组及IL—1α组比较分化率更加显著(P<0．05)。结论 BMSCs在体外培养条件下,联合GDNF 和IL-1α诱导分化并配合使用高浓度血清(10％)可获得神经巢蛋白阳性的神经前体细胞及表达D2受体阳性的多巴胺能神经元;骨髓源性神经干细胞可以诱导分化为多巴胺能神经元。     

慢病毒转染胶质细胞源性神经营养因子基因的胎儿骨髓间充质干细胞的研究

目的 探讨胎儿骨髓间充质干细胞(fBMMSCs)经慢病毒转染胶质细胞源性神经营养因子(GDNF)基因后GDNF的表达情况.方法 利用贴壁培养法,从流产胎儿的股骨骨髓中分离培养fBMMSCs,并进行扩增.取第5代fBMMSCs,用慢病毒载体转染GDNF基因.应用免疫组化法及ELISA法检测被转染的fBMMSCs GDNF的表达情况.结果 被转染GDNF基因的fBMMSCs GDNF表达阳性,且培养液中GDNF含量随培养时间延长而增高.结论 被慢病毒转染GDNF基因的fBMMSCs能表达GDNF.     

外源性SDF-1α和GDNF基因在恒河猴骨髓基质细胞内的稳定表达

目的 构建携带人基质细胞衍生因子-1α(SDF-1α)和胶质细胞源神经营养因子(GDNF)基因的真核表达质粒pBudCE4.1-SDF-1α-GDNF,转染恒河猴骨髓基质细胞(BMSCs),观察外源性SDF-1α和GDNF基因在BMSCs中的表达情况. 方法 应用基因重组技术,从cDNA文库获得GDNF和SDF-1α基因,重组到pBudCE4.1双表达载体上.pBudCE4.1-SDF-1α-GDNF经脂质体转染培养的恒河猴BMSCs,48 h后行GDNF和SDF-1α Western blot和免疫组化检测. 结果 酶切、PCR和DNA序列鉴定均证实插入基因片段的正确性;48 h后行West blot和免疫组化证实GDNF和SDF-1α在细胞内能有效表达.pBudCE4.1-GDNF-SDF-1α转染BMSCs表达的GDNF和SDF-1α蛋白的量是阴性对照细胞的10倍和6倍. 结论 双基因真核表达质粒DBudCE4.1-SDF -1α-GDNF转染至恒河猴BMSCs后,SDF-1α和GDNF基因在BMSCs内能有效表达,为BMSCs携带SDF-1α和GDNF双基因自体移植治疗相关疾病奠定了基础.     

KLF7对骨髓间充质干细胞增殖和施万样细胞分化的影响

目的探讨转录因子KLF7对骨髓间充质干细胞(BMSCs)增殖和施万样细胞分化的作用。方法 BMSCs培养与鉴定,应用腺病毒AAV-KLF7和AAV-NC转染BMSCs。将BMSCs分为AAV-KLF7组、AAV-NC组和正常组。应用Western blots和实时荧光定量PCR(qRT-PCR)检测KLF7及其靶基因神经生长因子(NGF)表达水平,EdU免疫荧光染色及MTT法检测各组细胞增殖情况,应用施万细胞分化培养液和免疫荧光染色检测各组细胞诱导分化为类施万样细胞情况。结果与正常组和AAV-NC组比较,AAV-KLF7组KLF7蛋白和mRNA表达水平显著升高,NGF mRNA表达亦显著升高(P0.05);AAV-KLF7组EdU阳性细胞比率显著增多,增殖活性显著增加(P0.05);诱导10 d后AAV-KLF7组S100阳性细胞比率显著增多(P0.05)。结论 KLF7显著促进BMSCs增殖和施万样细胞分化。     

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.     

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.     

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.     

《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。     

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.     

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。     

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.