创伤性脑损伤大鼠海马蛋白质差异表达研究

目的 应用蛋白质组学方法分析研究大鼠创伤性脑损伤后海马组织差异表达蛋白质,为筛选脑损伤早期诊断的生物学标志物提供理论依据.方法 采用双向凝胶电泳分离总蛋白,硝酸银染色,PDQuest 7.0图像分析软件分析获得的差异蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)对检测到的差异蛋白质点进行鉴定,获得肽质量指纹图谱,查阅蛋白质数据库得到所需蛋白质相关信息,分析大鼠创伤性脑损伤后4 h、8 h、12 h、24 h和48 h海马组织中差异表达蛋白质,每组样品电泳重复3次,每帧图谱检测到1800余个蛋白质点;选择2个背景清晰、重复性及分辨力良好、在各组表达差异明显并呈现时序性变化的蛋白质点进行质谱分析.结果 经双向凝胶电泳分离共获得18帧图,每组3帧,其中对照组及损伤后12 h组蛋白质图谱显示分离效果良好,蛋白质点清晰,点染色程度较深,数目较多.横纹和纵纹较少,图谱平均匹配率>80%.采用PDQuest 7.0图像分析软件对蛋白质图谱进行定量分析证实,相对分子质量为58.00×103的差异蛋白质为葡萄糖调节蛋白(葡萄糖调节蛋白58),在损伤后4 h、8 h和12 h表达水平上调;相对分子质量为28.40×103的差异蛋白质为蛋白酶体α亚单位3型.在损伤后4 h、8 h、12 h、24 h和48 h表达水平均上调.结论 葡萄糖调节蛋白58和蛋白酶体α亚单位3型可能与创伤性脑损伤的发生、发展密切相关,此为探讨脑损伤的发病机制提供了重要线索.     

颅脑创伤后亚低温脑保护的蛋白质组研究

目的 本研究应用差异蛋白质组学技术,探讨亚低温和常温条件下,创伤性颅脑损伤大鼠海马组织蛋白质的表达变化.方法 采用侧方液压冲击装置,建立大鼠中度脑损伤模型,亚低温组(n=3)于伤后维持体温(33±0.5)℃持续3h,常温组(n=3)始终维持体温(37±0.5)℃.取大鼠海马组织,通过差异荧光双向凝胶电泳、分离蛋白,获得二维的蛋白质分离图谱,然后通过胶内酶切、抽提酶解肽段、基质辅助激光解吸飞行时间质谱分析差异的蛋白质点,鉴定出变化的蛋白质.结果 通过DeCyder5.0(GE Healthcare)软件分析报告发现差异1.5倍以上的蛋白点17个(P＜0.05).通过对这些蛋白考染点进行质谱鉴定,鉴定出14个蛋白质,2个为同一种蛋白,实际差异蛋白数为13个.分别是细胞骨架蛋白、介导能量代谢的酶类、参与核酸合成、氧化应激反应的蛋白质、神经突触功能蛋白、细胞内信号传递蛋白及未知蛋白.结论 脑损伤后亚低温及常温条件下,大鼠海马组织蛋白质存在表达差异,鉴定出的差异蛋白质可能与亚低温保护效应的潜在作用机制有关.     

弱阳离子交换2蛋白质芯片检测大鼠闭合性颅脑损伤损伤后脑干差异蛋白

目的：研究大鼠闭合性颅脑损伤后脑干中蛋白质表达谱的变化及特点。方法：雄性SD大鼠48只,随机分为手术对照组及损伤后4h、8h、12h、24h和48h组,复制Marmarou’s落体打击脑损伤模型。取大鼠脑干进行常规HE染色,并采用蛋白质芯片检测差异蛋白,每只大鼠取100mg脑干,加1ml U9缓冲液和少量PMSF在冰上将组织匀浆,40000r/min离心2h,分离上清液并以50μl分装,冻存于-80℃,采用Bradford法检测样本蛋白质含量,对样本蛋白行WCX-2蛋白芯片研究,用蛋白质芯片阅读机进行蛋白质谱分析。结果：脑损伤后脑干组织部分神经细胞发生固缩坏死,WCX-2蛋白芯片研究发现与对照组相比,脑损伤后脑干中有2个蛋白质的表达谱发生改变。其中差异蛋白4158Da在对照组和4h、8h组几乎不表达,损伤后12h、24h和48 h组表达增加（P<0.01）;差异蛋白7862 Da在对照组高表达,损伤后12h、24h组表达降低（P<0.05）。结论：脑损伤可引起脑干中蛋白质表达谱发生变化。     

大鼠颅脑损伤后钠通道α亚单位1.3(Navl.3)在海马中的表达

目的 研究颅脑损伤(TBI)后钠通道α亚单位1.3(Navl.3)的mRNA和蛋白在海马中的表达情况.方法 对成年SD大鼠实施脑液压伤后,在伤后2 h、12 h、24 h和72 h处死,取伤侧海马行荧光定量PCR和Western blot检测Navl.3的mRNA和蛋白表达情况,通过免疫荧光染色检测Navl.3在海马的表达特点.结果 大鼠脑液压伤后Nayl.3的mRNA表达显著上调(P＜0.01).伤后12 h其上调达最高水平,而Navl.3蛋白的表达也在相同时间段出现显著上调(P＜0.01).免疫荧光染色显示Nayl.3在海马主要表达于神经元细胞.结论 TBI可导致Nayl.3的mRNA和蛋白表达显著上调,这可能是TBI后神经元细胞膜上钠通道功能异常及其诱发兴奋性毒性作用的分子学基础之一.     

大鼠颅脑损伤后钠通道α亚单位1.3（Nav1.3）在海马中的表达

目的研究颅脑损伤（TBI）后钠通道α亚单位1.3（Nav1.3）的mRNA和蛋白在海马中的表达情况。方法对成年SD大鼠实施脑液压伤后,在伤后2h、12h、24h和72h处死,取伤侧海马行荧光定量PCR和Western blot检测Nav1．3的mRNA和蛋白表达情况,通过免疫荧光染色检On,0Nav1．3在海马的表达特点。结果大鼠脑液压伤后Nav1.3的mRNA表达显著上调（P〈0.01）,伤后12h其上调达最高水平,而Nay1.3蛋白的表达也在相同时间段出现显著上调（P〈0．01）。免疫荧光染色显示Nav1.3在海马主要表达于神经元细胞。结论TBI可导致Nav1.3的mRNA和蛋白表达显著上调,这可能是TBI后神经元细胞膜上钠通道功能异常及其诱发兴奋性毒性作用的分子学基础之一。     

发育期大鼠内侧颞叶癫痫模型丙戊酸治疗前后的蛋白质组学研究

目的 研究丙戊酸(VPA)对发育期大鼠内侧颞叶癫痫(MTLE)模型海马蛋白质表达谱的影响,寻求与VPA抗癫痫作用相关的蛋白质以及探讨VPA在MTLE发病早期发挥抗癫痫作用的机制.方法 建立氯化锂-匹罗卡品诱导的发育期大鼠MTLE模型,采用二维凝胶电泳(2-DE)技术分离海马总蛋白,PDQuest软件识别差异表达蛋白质,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)进行鉴定.结果 建立了VPA治疗前后的MTLE模型大鼠海马蛋白质的2-DE图谱.质谱鉴定出9个差异表达的蛋白质,其中4个在VPA治疗后表达上调;5个下调.结论 本实验成功建立了发育期大鼠MTLE模型VPA处理前后的海马蛋白质2-DE图谱;β-synuclein、Neurogranin等差异表达蛋白可能与VPA在MTLE发病早期发挥抗癫痫作用的机制相关.     

大鼠全脑缺血再灌注损伤后海马自噬相关蛋白Beclin 1的表达变化

目的研究自噬相关蛋白Beclin 1在大鼠全脑缺血-再灌注损伤后海马中的表达情况,并探讨其意义。方法使用四动脉结扎法制作大鼠全脑缺血-再灌注损伤模型,实验动物随机分为：假于术组和缺血再灌注组。存全脑缺血15min后,分别再灌注0min、30min、3h、6h、12h、24h、1d、3d,使用Western blot检测各个时阃点大鼠全脑缺血-再灌后海马自噬相关蛋白Beclin1的表达情况。结果与假手术组比较,大鼠全脑缺血-再灌注损伤后1d,海马区Beclinl蛋白的表达最强（P〈0．05）。结论大鼠全脑缺血-再灌注损伤后海马自噬相关蛋白Beclinl表达上调,表明脑缺血-再灌注损伤后海马区域的自噬活性上调．     

大鼠颅脑损伤后亚低温治疗对Calpain及作用底物MAP-2表达的影响

目的探讨亚低温治疗对颅脑损伤后Calpain、MAP-2基因和蛋白表达的影响。方法将54只SD大鼠随机分为假手术组（n=6）、常温脑损伤组（n=24）和亚低温脑损伤组（n=24）。亚低温脑损伤组在液压打击伤后即接受持续4h的亚低温治疗。伤后6h、12h、24h和72h4个时间点分别处死3只常温脑损伤组和亚低温脑损伤组大鼠。荧光PCR、Western blot半定量检测皮质Calpain及MAP-2基因转录和蛋白的表达。结果颅脑损伤后12h及24h亚低温使Calpain mRNA表达增加（P〈0．05）,伤后6h、12h、24h和72h亚低温均可减少Calpain蛋白的升高,伤后12h及72h尤其显著（P〈0．05）。与假手术组比较,常温脑损伤组和亚低温脑损伤组MAP-2基因转录均减少（P〈0．05）;与常温脑损伤组比较,伤后6h、12h和24h亚低温可抑制MAP-2基因转录的下调,但亚低温脑损伤组MAP-2蛋白的表达均比同时间点常温组低（P〈0.05）。结论颅脑损伤后亚低温治疗的脑保护机制可能与调节Calpain蛋白的表达有关,而亚低温与MAP-2的关系还有待进一步研究。     

大鼠全脑缺血再灌注损伤后海马白噬相关蛋白LC3表达变化的研究

目的研究自噬微管相关蛋白轻链3（microtubule—associated protein 1 light chain3,LC3）在大鼠全脑缺血损伤后海马的表达情况,并探讨自噬在脑缺血损伤中的意义。方法使用四动脉结扎法制作大鼠全脑缺血模型再灌注损伤模型,实验动物随机分为：假手术组（sham组）、缺血再灌注纽。在全脑缺血15min后,分别再灌注0,30min,3,6,12,24h,1,3d,使用Westernblotting的方法检测各个时间点大鼠全脑缺血／复灌后海马LC3Ⅱ／LC3 Ⅰ蛋白的表达情况来研究脑缺血再灌注损伤后海马自噬现象的变化情况。结果与假手术组相比,大鼠全脑缺血再灌注损伤后3h,海马区LC3Ⅱ／LC3Ⅰ蛋白的表达开始上调,在全脑缺血再灌注损伤后1d表达最强（P〈0．05）。结论大鼠全脑缺血再灌注损伤后海马LC3Ⅱ／LC3Ⅰ蛋白表达上调,表明脑缺血再灌注损伤后海马区域的自噬活性上调。     

亚低温抑制大鼠弥漫性脑损伤后细胞凋亡的研究

目的探讨大鼠不同程度弥漫性脑损伤后脑组织的凋亡变化过程及亚低温治疗对脑细胞凋亡的抑制作用.方法采用大鼠Marmarou颅脑创伤装置制作弥漫性脑损伤模型,然后将128只Wistar大鼠分为未损伤组(对照组)、重度损伤组、轻度损伤组和亚低温治疗组.通过电子显微镜、组织切片原位末端标记DNA片段(TUNEL染色)、琼脂糖凝胶电泳(DNA Ladder法)等方法,观察和比较不同程度脑损伤后,大鼠脑皮层及海马区凋亡细胞的形态、特点和数量.结果(1)损伤后24～48 h,皮层及海马区可见大量细胞皱缩、核碎裂、核不规则等细胞凋亡现象,48 h较24 h更为严重;亚低温治疗后24～48 h,电子显微镜观察皮层及海马区未见细胞皱缩、核碎裂等细胞凋亡现象.(2)TUNEL染色结果显示,随着损伤程度的加重凋亡明显加重,损伤后48 h达高峰,然后逐渐下降.轻度损伤组细胞凋亡主要限于海马CA2和CA3区;重度脑损伤组细胞凋亡涉及整个海马结构,同时还广泛累及额顶区皮质.损伤后第24、48、72 h,皮层及海马区的凋亡细胞数量较同期未治疗组明显减少.(3)重度损伤后48 h,海马和皮层区细胞琼脂糖电泳可见典型的DNA梯状带,其他时间未见梯状带.亚低温治疗组、轻度脑损伤组及未损伤组亦未见梯状带.结论轻度弥漫性脑损伤后,脑细胞凋亡多发生于海马CA2和CA3区;重度脑损伤后皮层及海马区细胞可发生广泛凋亡.细胞凋亡随着损伤程度的加重而加重,高峰位于伤后第2 d.亚低温治疗可有效地抑制大鼠弥漫性脑损伤后的细胞凋亡.     

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     

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.     

广西农村壮族妇女精神分裂症患者生活质量状况调查

目的研究农村壮族妇女精神分裂症患者的生活质量及影响因素。方法前瞻性的队列研究。采用随机分层抽样法分为农村壮族妇女精神分裂症组、农村汉族妇女精神分裂症组、农村正常妇女对照组,应用“世界卫生组织生存质量测定报告”(WHOQOL-100)及PANSS量表调查其生活质量和疾病的严重程度。结果农村壮族妇女精神分裂症患者生活质量明显低于农村汉族妇女精神分裂症患者,影响其生活质量的相关因素是生活环境及精神支柱/个人信仰。结论经济贫困、环境条件、缺乏有效的医疗服务和社会保障是农村壮族妇女精神分裂症患者生活质量低的关键。因此,建立农村壮族社区精神卫生服务网络势在必行。     

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.     

《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。     

The origins of innervation of the esophagus of the dog

This study defined the origins of extrinsic efferent and afferent innervation of the normal canine esophagus. When all the layers of the wall of the 3 esophageal regions (cervical, thoracic and abdominal) were injected with horseradish peroxidase (HRP), labeled nerve cells were found in the nucleus ambiguus (NA) and parasympathetic nucleus of X (PX) of the brainstem. Most labeled cells in the NA were located in the compact column (retrofacial nucleus) while labeled cells in the PX were located in separate rostral and caudal areas. There was no somatotopic organization in either the NA or PX. Labeled sympathetic postganglionic neurons were found in the cranial cervical, middle cervical, cervicothoracic, thoracic sympathetic trunk and celiacomesenteric ganglia. The HRP injection of the esophageal wall labeled sensory cell bodies in the glossopharyngeal, proximal and distal vagal, and C2-T6 spinal ganglia. There was no discernible pattern of distribution of labeled cells in the autonomic or sensory ganglia. When the HRP injections were confined to the mucosa-submucosa layers of the thoracic esophagus, a small number of labeled cells were identified in the NA; however, no labeled cells were found in the NA when injections were confined to the mucosa-submucosa of either the cervical or abdominal esophageal regions. With these confined injections, the labeled nerve cells appeared in the rostral part of the PX. Thus, it appeared that the internal tunics of the esophagus (i.e., the mucosa and submucosa) were innervated by neurons in the rostral PX while the muscular tunic was innervated by neurons in the caudal PX and the rostral NA. After mucosa-submucosa injections, labeled sympathetic neurons appeared in the same ganglia that were identified after whole wall injections and these had a similar random distribution. These injections also labeled neurons in the glossopharyngeal, proximal vagal, and distal vagal ganglia, but unlike the whole wall injections there was no labeling in the spinal ganglia. This suggested that the labeled cells of the spinal ganglia seen after whole wall injections conveyed impulses from the tunica muscularis and serosa.     

Impact of our understanding of the genetic aetiology of epilepsy

A genetic contribution to aetiology is estimated to be present in up to 40% of patients with epilepsy. It is useful to categorise genetic epilepsies according to the mechanisms of inheritance into Mendelian disorders, non-mendelian or ‘complex’ disorders, and chromosomal disorders. Over 200 Mendelian diseases include epilepsy as part of the phenotype, and the genes for a number of these have been identified recently. These include autosomal recessive progressive myoclonic epilepsies such as Unverricht-Lundborg disease, Lafora disease and the neuronal ceroid lipofuscinoses, and three autosomal dominant idiopathic epilepsies. The last named have been shown to arise from mutations in ion channel genes. Autosomal dominant nocturnal frontal lobe epilepsy is caused by mutations in CHRNA4, benign familial neonatal convulsions by mutations in KCNQ2 and KCNQ3, and generalised epilepsy with febrile seizures plus by mutations in SCN1B. ‘Complex’, familial epilepsies are more difficult to analyse, but evidence has been obtained for loci predisposing to juvenile myoclonic epilepsy on chromosome 6p and 15q. Lastly, the genes underlying several spike-wave epilepsies in mice have been cloned, and three of these encode sub-units of voltage-gated calcium channels. Received: 29 September 1999/Accepted: 7 December 1999