去卵巢大鼠胆碱能神经系统改变及雌激素保护作用机制的研究

目的观察去卵巢大鼠胆碱能神经系统的改变及雌激素对其影响。方法利用免疫组织化学染色结合图像分析方法观察去卵巢大鼠海马CA1区胆碱乙酰转移酶(ChAT)的变化;利用RT—PCR方法检测去卵巢大鼠基底前脑神经生长因子(NGF)mRNA表达的变化。结果去卵巢大鼠海马CA1区ChAT的积分光密度(OD)值降低,基底前脑NGFmRNA平均表达水平明显降低,补充雌激素后以上各项指标均明显提高。结论(1)去卵巢大鼠海马CA1区胆碱能神经元ChAT活性下降,基底前脑胆碱能神经元NGFmRNA表达水平降低。(2)雌激素能够上调去卵巢大鼠NGFmRNA表达．而NGF能够增强ChAT的活性。促进乙酰胆碱(Ach)的合成和释放,改善学习记忆能力。     

雌激素对双侧穹隆-海马伞切断及去势大鼠脑源性神经营养因子表达的影响

目的　采用双侧穹隆-海马伞切断及双侧卵巢切除方法制作大鼠认知障碍模型,观察4个脑区脑源性神经营养因子(BDNF)表达的变化,以及补充外源性雌激素对BDNF的影响。方法选择健康雌性Wistar大鼠30只,先按2×2方差分析设计分别建立4个模型组:穹隆-海马伞切断组(FF组)、卵巢切除组(OVX组)、卵巢切除加穹隆-海马伞切断组(OF组)和对照组,然后再建立OF组同时给予雌二醇(E2 )治疗组(OF+E2 组)和双侧穹隆-海马伞切断加卵巢切除对照组,每组5只。E2 剂量为1mg/kg,每7天1次皮下注射,共4次28天。观察各组大鼠海马CA1区、皮质区、杏仁复合体区、基底前脑Meynert核区BDNF蛋白表达的变化。结果　(1)经F分析,双侧卵巢切除和双侧穹隆-海马伞切断与否存在主效应和显著交互效应(P<0 001)。FF组、OVX组和OF组的BDNF阳性细胞计数在海马CA1区、皮层、杏仁复合体、Meynert核均明显少于对照组(P<0 001),其中OF组在Meynert核区的BDNF阳性细胞计数少于FF组和OVX组(P<0 01); (2)OF组+E2 组在4个脑区的BDNF阳性细胞数均多于OF对照组(均P<0 01和<0 05)。结论　双侧穹隆-海马伞切断和(或)双侧卵巢切除均可导致大鼠脑内BDNF表达减少,补充外源性雌激素可显著增加BDNF的表达。     

铝致阿尔茨海默病模型大鼠卵巢结构功能与端脑雌激素受体亚型表达量的变化及补肾填精方的防治效应

目的　探讨慢性铝中毒所致阿尔茨海默病模型大鼠卵巢组织细胞结构、血清雌激素水平及端脑皮质雌激素受体亚型表达量的变化及补肾填精方的防治效应。方法　用放射免疫法测定血清雌二醇水平 ,以 HE染色法观察卵巢组织细胞结构和以免疫组化法测定端脑雌激素受体 α和 β亚型的表达量。结果　模型大鼠血清雌二醇水平显著降低 (P <0 .0 0 0 1 ) ,卵巢组织细胞萎缩变性和脱落 ,端脑皮质雌激素受体 α亚型表达明显减弱 ,β亚型表达明显增强。补肾填精方干预后 ,大鼠血清雌二醇水平没有明显变化 ,卵巢组织细胞结构明显好转 ,端脑皮质雌激素受体α表达明显增强及β表达明显降低。结论　慢性铝中毒后阿尔茨海默病模型大鼠卵巢功能减退和端脑皮质雌激素受体亚型表达量异常 ,可能与模型大鼠学习记忆能力降低有关 ,补肾填精方能抑制上述效应但不升高血清雌二醇水平。     

骨形态发生蛋白4在Aβ诱导的神经毒性病变中的表达变化

目的探讨骨形态发生蛋白4(bone morphogenetic protein 4,BMP4)在β淀粉蛋白(amyloidβ,Aβ)诱导的神经毒性病变中的变化。方法 SD大鼠海马神经元原代培养,经过不同浓度(10、15、20μmol/L)Aβ25-35毒性片段处理后,通过定量PCR技术检测BMP4mRNA表达水平。SD大鼠双侧基底前脑注射Aβ1-40建立大鼠痴呆模型,通过免疫组化及免疫印迹检测基底前脑及海马区BMP4蛋白表达水平。结果 Aβ25-35处理组海马神经元内BMP4mRNA表达较空白对照组明显减少(P0.05)。免疫组化和免疫印迹实验结果显示,与对照组比较,Aβ1-40注射组大鼠基底前脑区BMP4蛋白表达减少,海马区其表达升高(P0.05)。结论 Aβ直接下调神经元内BMP4转录及表达,不同脑区内BMP4蛋白可能存在负反馈调节作用。     

雌激素对阿尔茨海默病的干预作用

目的选择绝经期(65岁前)的轻中度AD(阿尔茨海默病)妇女,给予雌激素替代治疗(HRT),于治疗前、后分别测试其智力的变化;以双侧穹隆-海马伞切断制作AD大鼠模型,并给予雌激素治疗,观察穹隆-海马伞切断组、雌激素治疗组避暗实验的错误次数及潜伏期的变化,及脑内海马区(CA1区)烟碱型乙酰胆碱受体 (nAchR)表达的变化;从临床和基础两方面观察雌激素对AD的干预作用,探讨雌激素的作用及相关机制。方法选择21例绝经期(65岁前)的轻中度AD妇女,随机双盲比较治疗,用老年痴呆认知能力评价表,对治疗前、治疗后 16周分别进行智能评分。健康雌性Wistar大鼠,随机分为穹隆-海马伞切断组和雌激素治疗组;每组大鼠5只。在脑立体定位仪上切断双侧穹隆-海马伞,建立模拟AD的动物模型,对比穹隆-海马伞切断组和雌激素治疗组大鼠的智力变化;并应用免疫组织化学技术观察大鼠脑内CA1区nAchR细胞表达的变化。结果 21例患者行雌激素替代疗法后,修订长谷川量表(HDS-R)及精神认识能力(CCSE)评分比治疗前显著提高(P<0．001,P<0．01),社会活动功能调查(FAQ)评分比治疗前显著降低(P<0．05)。与穹隆-海马伞切断组大鼠比较,雌激素治疗组大鼠避暗实验的错误次数明显减少,潜伏期显著延长(P<0．05),脑内nAchR的表达显著增加(P<0．01)。结论临床和基础实验均证实雌激素能提高AD的智力,并认为其与脑内nAchR表达的增加有关。     

SMAD6在Aβ诱导的神经毒性病变中的表达变化

目的探讨母抗Dpp小同源物6(small mothers against decapentaplegic homolog 6,SMAD6)在β淀粉样蛋白(amyloidβ,Aβ)诱导的神经毒性病变中的表达变化。方法取孕15~17d的SD大鼠的胎鼠,进行海马神经元原代培养,经不同浓度(10、15、20μmol/L)Aβ25-35毒性片段处理后,通过定量PCR技术检测SMAD6mRNA表达水平。取2月龄SD雄性大鼠12只,随机分为Aβ注射组和空白对照组,每组6只。采用双侧基底前脑注射Aβ1-40建立大鼠痴呆模型,通过免疫组化及免疫印迹检测基底前脑及海马区SMAD6蛋白表达水平。结果不同浓度Aβ25-35处理组海马神经元内SMAD6mRNA表达均较对照组减少(F=602.1,P0.01)。免疫组化结果显示,与对照组比较,Aβ1-40注射组大鼠基底前脑区SMAD6蛋白表达减少(68103±1520比96036±1804;t=20.51,P0.01),而海马区其表达增多(96441±1852比55039±1528;t=29.87,P0.01);免疫印迹结果显示,与对照组比较,Aβ1-40注射组大鼠基底前脑区SMAD6蛋白表达减少(0.1042±0.0067比1.000±0.0986;t=15.69,P0.01),而海马区其表达增多(12.5525±2.6803比1.000±0.0135;t=7.465,P0.01)。结论 Aβ可直接下调神经元内SMAD6mRNA转录及表达,不同脑区内SMAD6蛋白表达可能受负反馈调节机制影响。     

大鼠脑损伤后NMDAR1表达及其认知障碍变化的关系

目的同步考量脑损伤后大鼠与认知功能较为密切的脑区N-甲基-D-天冬氨酸受体1（NMDAR1）表达与认知功能的变化。方法参照Feeney法建立大鼠创伤性脑损伤模型,伤后1d,2d,4d,7d1）．2免疫组化SABC法检测大鼠额叶皮质、海马区、基底前脑区NMDAR1表达,以行走实验、平衡实验及记忆功能测定评估大鼠认知障碍变化。结果轻、中型脑损伤组大鼠于伤后2d认知障碍最严重,分别于伤后3,7d基本恢复正常。轻型、中型脑损伤组脑额叶皮质、海马区、基底前脑区NMDAR1均于伤后1d升高,于2d降至较低水平后再呈缓慢增高趋势,与认知障碍变化趋势呈同步变化,且中型脑损伤组NMDAR1表达高于假手术组与轻型脑损伤组（P〈0．05）。结论大鼠经创伤性脑损伤后,额叶皮质、海马区、基底前脑区细胞中的NMDAR1含量和损伤后认知障碍的变化趋势有相似性;创伤性脑损伤后表达增加的NMDAR1对大鼠认知功能有加重损害作用。     

雌激素受体β对阿尔茨海默病大鼠海马内炎症反应及淀粉样β蛋白的沉积的影响

目的研究雌激素受体β(ERβ)对阿尔茨海默病大鼠海马内炎症反应及淀粉样β蛋白沉积的影响,并探讨其可能的机制。方法β淀粉样蛋白脑室内注射建立阿尔茨海默病大鼠模型,并将模型分为对照组、ERβ组和shRNA组。对照组不做任何处理,ERβ组和shRNA组大鼠分别通过脑室内注射载有雌激素受体β基因或shRNA的慢病毒质粒。通过Morris水迷宫检测各组大鼠行为学,Western blot检测各组大鼠脑组织内Akt及β淀粉样蛋白表达水平,qRT-PCR检测TNF-α和IL-1βmRNA表达,并通过免疫荧光染色检测Aβ的表达。结果ERβ组大鼠记忆能力明显高于对照组和shRNA组大鼠(P<0.05),ERβ组大鼠海马内Akt含量明显高于对照组和shRNA组大鼠(P<0.05),并且ERβ组大鼠海马内IL-1和TNF-αmRNA以及β淀粉样蛋白含量明显低于对照组和shRNA组大鼠(P<0.05)。结论过表达ERβ在不依赖雌激素的情况下可减轻AD大鼠海马内炎症反应和β淀粉样蛋白的沉积,改善AD大鼠行为学的变化,其神经保护作用可能是通过激活Akt途径实现。     

切除卵巢对大鼠学习记忆和海马雌激素受体α的影响及电针的干预作用

目的 研究电针对去卵巢大鼠行为学及海马区雌激素受体α(estrogen receptor alpha,ERα)蛋白和mRNA表达的影响.方法 将40只雌性SD大鼠随机分为4组:假手术组、模型组、假电针组、电针组,各10只.采用卵巢切除大鼠模型,造成低雌激素记忆障碍,去势2周后进行电针刺激,连续治疗3个月.以Morris水迷宫测试空间学习记忆能力,酶联免疫吸附分析(enzyme-linked immunosorbent assay,ELISA)检测血清雌二醇(estradiol,E2)浓度,免疫印迹法(Western blot,WB)和实时荧光定量PCR分别检测海马区ERα蛋白和mRNA的相对表达量.结果 模型组、假手术组、电针组和假电针组大鼠逃避潜伏期分别为(49.65±3.82)s、(15.26±3.37)s、(14.33±3.62)s、(25.46±2.87)s,游泳距离分别为(941.88±37.77) cm、(291.66±23.42) cm、(261.52±28.76)cm、(471.01±33.31)cm,跨越平台数分别为(2.35±0.44)、(8.94±0.91)、(8.53±0.83)、(3.64±0.88)次,血清E2浓度分别为(38.16±5.44) pg/mL、(65.13±7.72) pg/mL、(56.47±6.70) pg/mL、(49.29±5.90)pg/mL,海马ERa蛋白表达分别为(0.406±0.040)、(0.985±0.086)、(0.796±0.072)、(0.498±0.063),海马ERamRNA表达分别为(1.037±1.021)、(2.303±0.065)、(1.410±0.062)、(1.153±0.035),模型组较假手术组其逃避潜伏期和游泳距离延长,跨越平台数增加,血清E2浓度及海马ERa蛋白和mRNA表达均降低,电针组和假电针组均较模型组有所改善,但电针组改善更明显,上述差异均有统计学意义(均P＜ 0.01).结论 电针能够提高去卵巢大鼠学习记忆能力,其机制可能通过升高体内雌激素浓度上调海马ERα蛋白和mRNA的表达有关.     

大鼠脑缺血再灌注后大脑海马区自噬的研究

目的检测大鼠脑缺血再灌注后海马神经细胞损伤及自噬变化,并比较海马CA_1及CA_3区的自噬情况。方法用线栓法制作大鼠脑局部缺血再灌注模型,实验动物随机分为:对照组(Control组)、假手术组(Sham组)和大脑中动脉栓塞再灌注组(MCAO组)。HE染色检测大鼠海马神经细胞损伤。透射电镜下观察海马神经元内自噬小体。免疫荧光法检测自噬相关蛋白LC3、Beclin-1的表达水平。结果与Control组及Sham组比较,大鼠脑缺血再灌注后,海马神经细胞损伤严重;海马神经元内有自噬体形成;自噬标记物LC3、Beclin-1蛋白表达水平显著增加,且CA_1区的表达量明显高于CA_3区(差异均具有统计学意义P0.05)。结论脑缺血再灌注可激活海马神经细胞内的自噬,并进一步引起细胞损伤;模型动物海马CA_1区的自噬水平显著高于CA_3区,暗示CA_1区对脑缺血损伤具有较高的敏感度。     

The neuroendocrinology of stress and the pathophysiology and therapy of depression and anxiety

Clinical and preclinical studies have gathered substantial evidence that stress response alterations play a major role in the development of major depression, panic disorder, and post-traumatic stress disorder. The stress response, the hypothalamic pituitary adrenocortical (HPA) system and its modulation by corticotropin-releasing hormones (CRH),corticosteroids,and their receptors, and the roles of natriuretic peptides and neuroactive steroids are described. We review the role of the HPA system in major depression, panic disorder, and post-traumatic stress disorder and its possible relevance for treatment. Impaired glucocorticoid receptor function in major depression is associated with an excessive release of neurohormones such as CRH, to which a number of signs and symptoms characteristic of depression can be ascribed. In panic disorder, a role of central CRH in panic attacks has been suggested. Atrial natriuretic peptide (ANP) is causally involved in sodium lactate-induced panic attacks. Furthermore, preclinical and clinical data on its anxiolytic activity suggest that nonpeptidergic ANP receptor ligands may be potentially useful in the treatment of anxiety disorders. Post-traumatic stress disorder is characterized by a peripheral hyporesponsive HPA system and elevated CRH concentrations in the CSF. This dissociation is probably related to an increased risk of this disorder. We further review recent data that describe an important role of GABA(A)-receptor modulatory,3 alpha-reduced neuroactive steroids in major depression, anxiety, and its treatment. Antidepressants are effective in both depression and anxiety disorders and have major effects on the HPA system,especially on glucocorticoid and mineralocorticoid receptors. Normalization of HPA system abnormalities is a strong predictor of the clinical course, at least in major depression and panic disorder. Currently,CRH-R1 or glucocorticoid receptor antagonists and ANP receptor agonists are being studied and may provide future treatment options more closely related to the pathophysiology of these disorders.     

Polymorphisms of DRD4 and DRD3 and risk of avoidant and obsessive personality traits and disorders

We investigated whether polymorphisms of the dopamine D4 receptor (DRD4) and polymorphisms of the dopamine D3 receptor (DRD3) were associated with personality disorder symptomatology rather than with personality traits such as novelty seeking. DNA was obtained from 145 depressed patients in a clinical trial. These patients were assessed for the presence of personality disorder symptoms and disorders. The 2-repeat allele of the DRD4 exon III polymorphism was associated with increased rates of avoidant and obsessive personality disorder symptomatology. The T,T genotype of the DRD4 -521 C>T polymorphism was also associated with increased rates of avoidant and obsessive personality disorder symptomatology. The Gly9,Gly9 genotype of the DRD3 Ser9Gly polymorphism was associated with increased rates of obsessive personality disorder symptomatology. None of these three polymorphisms were associated with novelty seeking or other temperament traits on the Temperament and Character Inventory. Our results suggest that genetic polymorphisms of DRD4 and DRD3 may well be associated with personality traits, and that conflicting findings to date may arise from the problem of phenotype definition.     

沙盘游戏疗法在地中海贫血患儿心理干预中的应用

本文目的是对沙盘游戏疗法在地中海贫血患儿心理干预中的应用进行综述,以期为地中海贫血患儿的心理康复提供参考。地中海贫血是以珠蛋白生成障碍为主要特征的遗传性疾病,由于长期输血治疗,患儿存在较多的心理和行为问题。沙盘游戏疗法作为一种有效、实用的儿童心理治疗方法,对提高地中海贫血患儿的康复效果、改善生存质量有重要的临床意义。     

癫痫共病抑郁的机制及临床诊疗

本文目的是探讨癫痫共病抑郁的可能机制及临床诊疗。癫痫是一种常见的、慢性的、致残性的神经疾病,癫痫患者生活质量下降,存在明显的负性情绪,常伴发各种精神疾病。癫痫与抑郁具有共同的神经生物学基础,可能存在共同的发病机制。本文从癫痫共病抑郁的发病机制、临床诊断及治疗方面予以总结归纳。     

Efficacy and Effectiveness of Child and Adolescent Psychotherapy and Pharmacotherapy

Decades of intervention research have produced a rich body of evidence on the effects of psychotherapies and pharmacotherapies with children and adolescents. Here we summarize and critique that evidence. We review findings bearing on the efficacy of psychosocial treatments and medications under controlled experimental conditions. We also report evidence, where available, on the effectiveness of both classes of treatment with clinically referred youth treated in real-world clinical contexts. In general, the large body of evidence on efficacy contrasts sharply with the small base of evidence on effectiveness. Addressing this gap through an enriched research agenda could contribute importantly to linking scientific inquiry and clinical practice—to the benefit of both ventures. This is one element of a multifaceted agenda for future research and for synthesis of research, which will require the interplay of multiple disciplines related to child and adolescent mental health.     

Seclusion and restraint and prediction of violence

The authors studied the use of seclusion and restraint on an inpatient unit in a state psychiatric hospital. Of 69 randomly selected inpatients, 51% experienced seclusion or restraint at least once. More psychotic than nonpsychotic patients required seclusion or restraint. However, neither psychosis/nonpsychosis nor voluntary/involuntary admission status predicted the likelihood of violent threats or actions. Patients experiencing seclusion and restraint showed a nonsignificant trend toward longer mean length of stay in the hospital. The frequency of patient behavior leading to seclusion or restraint appeared to be directly related to the stimulation caused by the presence of many staff members and other patients.     

Comparison of trigeminal and spinal modulation of pain and nociception

Modulation of pain and nociception by noxious counterstimulation, also called "diffuse noxious inhibitory controls" or DNIC-like effect, is often used in studies of pain disorders. It can be elicited in the trigeminal and spinal innervation areas, but no study has previously compared effects in both innervation areas. Therefore, we performed a study comparing DNIC-like effects on the nociceptive flexion reflex (NFR) and the nociceptive blink reflex as well as the respective pain sensations. In 50 healthy volunteers, the blink reflex elicited with a concentric electrode and the NFR were recorded before and after immersion of the contralateral hand in cold water. Responses were recorded as the subjective pain sensation and the reflex size. The cold water immersion of the contralateral hand elicited a reduction of both subjective pain sensation and reflex amplitude following the stimulation of both reflexes. However, there were no strong correlations between the individual reductions of both subjective pain sensation and reflex amplitude for both reflexes, and neither when results of the two reflexes were compared with each other. The dissociation between DNIC-like effects on pain and on nociception, which had been found previously already for the NFR, implies that both effects need to be studied separately.