人载脂蛋白APOA5的克隆表达纯化及相互作用蛋白研究

人类的载脂蛋白A5（apolipoprotein A5，APOA5）是一个新近发现的载脂蛋白家族成员。它在血浆中的含量比其他载脂蛋白低1-2个数量级，但能显著影响血浆三酰甘油水平，对血脂代谢具有重要意义，可以作为降血脂药物治疗中一个强有力的潜在靶标。 由于APOA5在血浆中含量低，直接从血浆中分离纯化很困难，国内一直没有报道简易可靠的纯化方法。为进一步研究APOA5的生物学特性，探讨其与TG代谢中的其它关键成分之间的相互关系，揭示其在脂类代谢相关疾病中的重要地位，必须有大量的蛋白和抗体用于基础研究。因此本研究首先利用基因工程技术，诱导表达纯化APOA5蛋白，免疫动物制备多克隆抗体，为进一步研究人肝脏细胞中APOA5的相互作用蛋白，研究APOA5蛋白在肝脏细胞中的功能奠定基础。 为了深入研究APOA5在肝脏中如何行使功能，我们采用细菌双杂交技术寻找与APOA5相互作用的蛋白因子。并采用Pull-down技术，免疫荧光及免疫共沉淀技术进一步确证其在体外和体内的相互作用关系，为进一步阐明APOA5在体内的生理功能提供了新的线索。 第一部分 APOA5基因的克隆、原核表达、纯化及其多克隆抗体的制备 本研究首先应用基因克隆技术，从人肝癌细胞系SMMC-7721的cDNA中扩增出1.1 kb的ApoA5基因全长序列。然后将其克隆至表达载体pThioHisD，构建原核表达载体pTH-APOA5。该重组质粒转化至大肠杆菌 BL21（DE3），成功实现人APOA5融合蛋白在大肠杆菌中的表达。经发酵得到高效表达的融合蛋白。 融合蛋白在 IPGT 诱导下以包涵体的形式大量表达。利用融合蛋白上的一段组氨酸序列，用镍离子亲和柱进行纯化和复性后，获得较高纯度的人APOA5融合蛋白。利用该融合蛋白免疫新西兰大耳白兔，获得了高效价的兔抗人APOA5多克隆抗体，Western Blot结果显示此多克隆抗体与APOA5特异性结合。 第二部分 细菌双杂交筛选与APOA5相互作用的蛋白 本实验首先构建了pBT-APOA5重组质粒，经双酶切、PCR和测序鉴定证明重组诱饵质粒构建成功，并进行了表达、自激活鉴定。Western Blot鉴定证实报告菌株中表达了分子量为 68 kD左右的重组融合蛋白，与预测的分子量APOA5（41 kD）/lamda cI （27 kD）一致。自激活实验证明诱饵蛋白不能单独激活报告基因，可用于筛选人肝脏cDNA文库。经过双重抗性筛选和回复筛选，分离出10个阳性克隆。对结果进行生物信息学分析，得到7个与APOA5相互作用的蛋白，其中BI1为细胞凋亡调节因子；ATP6、CYTB、ND2、COX-1为线粒体表达蛋白； ALB、TTR为血清蛋白。 第三部分 APOA5与BI1相互作用的确证 首先构建了BI1的原核表达载体pGEX-5X-3-BI1，利用Pull-down实验检测了APOA5与BI1在体外具有相互作用。然后构建了BI1的真核表达载体pCDNA3.1-HA-BI1和APOA5的真核表达载体pCDNA3.1-APOA5，并验证其表达。通过免疫荧光细胞内共定位研究发现，靶蛋白APOA5主要分布于胞浆，与BI1在HEK293细胞有共定位，即APOA5与BI1存在相互作用的可能。最后利用免疫共沉淀手段，在HEK293细胞中确证了靶蛋白APOA5与BI1在体内的相互作用。 上述研究结果，为深入研究APOA5在体内的生物学功能提供了新的思路。 Apolipoprotein A5 (APOA5) is a newly discovered protein belongs to apolipoprotein family. APOA5’s concentration is 1-2 orders of magnitude lower than other apolipoproteins in the circulation. APOA5 significantly affected plasma triglyceride levels, which is important on lipid metabolism. APOA5 has strong potential to be used as a hypolipidemic drug target. Large amount of APOA5 protein and antibodies are needed in basic research, such as biological characteristics study of the APOA5, its relationship with other key components in TG metabolism, its role played in Lipid metabolism-related diseases. Due to its low concentration in plasma, separation and purification of APOA5 from the plasma is very difficult. Until now no report on simple and reliable method for purification has been published in China. In this study, we firstly got APOA5 recombinant protein using genetic engineering technology. The purified recombinant protein was used to immunize rabbits to get antiserum. It is important for further study of the APOA5 protein-interacting protein. And it lays the foundation for studing APOA5 function in liver. In order to study APOA5 function in liver, we used bacterial two-hybrid technology to find the APOA5 protein interactor. Pull-down, immunofluorescence and immunoprecipitation techniques were used to further confirm the interaction between APOA5 with its interactor in vitro and in vivo. All of these stdudies provided new clues on its physiological functions in vivo. Part I: Cloning, prokaryotic expression, purification and polyclonal antibody preparation of APOA5 First of all, we amplified APOA5 CDS sequence from the human hepatoma cell line SMMC-7721, and subcloned into Expression vector pThioHisD, and got the recombinants named pTH-APOA5. The plasmid was transformed to BL21 (DE3). E. coli BL21(DE3) cells bearing the pTH-APOA5 plasmid were cultured and APOA5 protein synthesis was induced by the addition of IPTG. Recombinant protein was expression in the form of inclusion. Inclusion bodies were dissolved in phosphate-buffered saline containing 8 M urea and 40 mM imidazole, then applied to a Ni2+ affinity column, and were eluted in a buffer containing 4 M urea and 200 mM imidazole. Fractions containing the APOA5 protein were pooled and dialyzed against buffer containing phosphate-buffered saline. Antiserum to recombinant human APOA5 was generated by immuning rabbit. Western Blot showed that this antiserum specific binding with APOA5. Part II Two-hybrid system screening protein interactions with the APOA5 The coding sequence of human APOA5 was amplified using synthetic oligonucleotide primers from pTH-APOA5 vector and was subcloned into the pBT plasmidc to yield pBT-APOA5 vector. DNA sequencing was performed to verify that no unwanted mutations occurred during the process of plasmid vector construction. We verified recombinant protein expression and tested self-activation by pBT-APOA5 prior to screening. Western Blot verified inducing a 68 kD band, consistent with the predicted molecular weight (APOA5 41 kD, lamda cI 27 kD). pBT-APOA5 can be used for screening human liver cDNA library because it can not self-activation. Totally 10 positive clones were isolated. The nucleotide sequence of the positive clones were determined and compared to NCBI nucleotide sequence databases. We got 7 protein which interact with APOA5, included BI1(Apoptosis regulator); ATP6, CYTB, ND2, COX-1(Mitochondrial protein) and ALB, TTR(Serum protein). Part III Confirming of interaction between APOA5 with BI1 pGEX-5X-3-BI1 vector was subcloned at first. Pull-down experiments were used to detect the interaction between APOA5 with BI1 in vitro. Later, pCDNA3.1-HA-BI1 and pCDNA3.1-APOA5 were subcloned. Through immunofluorescence co-localization study, we found APOA5 mainly distributed in the cytoplasm. APOA5 is co-localization with BI1 in HEK293 cells. Finally, we verified interaction between APOA5 with BI1 in vivo through immunoprecipitation.

Capillary electrophoresis with electrochemiluminescence detection for measurement of aspartate aminotransferase and alanine aminotransferase activities in biofluids

A new sensitive assay for aspartate aminotransterase (AST) and alanine aminotransferase (ALT) activities in biofluids was developed, based on the separation and detection of alanine, glutamate, and aspartate using capillary electrophoresis (CE) with electrochemiluminescence (ECL) detection. The three amino acids were separated in 5 mM phosphate of pH 2.1 as background electrolyte, and detected on a 500 mu m platinum disk electrode at 1.2 V (versus Ag/AgCl) in the presence of 10 mM tris(2,2'-bipyridyl)ruthenium(II) dissolved in 80 mM phosphate of pH 10.5. A mass detection limit of 37.3 fmol (or 81.5 fmol) for glutamate, corresponding to the product in the enzyme reaction catalyzed by 1.24 x 10(-9) U AST (or 2.72 x 10(-9) U ALT) in a 30 min reaction period, was achieved. This assay was applied to investigate the cytotoxicity effect of ethanol on HepG2 cells and differentiating nonalcoholic steatohepatitis (NASH) from alcoholic liver disease, indicating that the technique is promising for the application in the cell biological and clinical fields.

DOPAMINE D-1 RECEPTOR MECHANISMS IN THE COGNITIVE PERFORMANCE OF YOUNG-ADULT AND AGED MONKEYS

Dopamine (DA) D-1 receptor compounds were examined in monkeys for effects on the working memory functions of the prefrontal cortex and on the fine motor abilities of the primary motor cortex. The D-1 antagonist, SCH23390, the partial D-1 agonist, SKF38393, and the full D-1 agonist, dihydrexidine, were characterized in young control monkeys, and in aged monkeys with naturally occurring catecholamine depletion. In addition, SKF38393 was tested in young monkeys experimentally depleted of catecholamines with chronic reserpine treatment. Injections of SCH23390 significantly impaired the memory performance of young control monkeys, but did not impair aged monkeys with presumed catecholamine depletion. Conversely, the partial agonist, SKF38393, improved the depleted monkeys (aged or reserpine-treated) but did not improve young control animals. The full agonist, dihydrexidine, did improve memory performance in young control monkeys, as well as in a subset of aged monkeys. Consistent with D, receptor mechanisms, agonist-induced improvements were blocked by SCH23390. Drug effects on memory performance occurred independently of effects on fine motor performance. These results underscore the importance of DA D-1 mechanisms in cognitive function, and provide functional evidence of DA system degeneration in aged monkeys. Finally, high doses of D-1 agonists impaired memory performance in aged monkeys, suggesting that excessive D-1 stimulation may be deleterious to cognitive function.

DOPAMINE D2 RECEPTOR MECHANISMS CONTRIBUTE TO AGE-RELATED COGNITIVE DECLINE - THE EFFECTS OF QUINPIROLE ON MEMORY AND MOTOR-PERFORMANCE IN MONKEYS

The D2 dopamine (DA) receptor agonist, quinpirole, was characterized in young adult monkeys, young reserpine-treated monkeys and aged monkeys to assess the contribution of DA to age-related loss of prefrontal cortical (PFC) cognitive function, Monkeys were tested on a delayed response memory task that depends on the PFC, and a fine motor task that taps the functions of the motor cortex, In young adult monkeys, low quinpirole doses impaired performance of the PFC and fine motor tasks, while higher doses improved memory performance and induced dyskinesias and ''hallucinatory-like'' behaviors. The pattern of the quinpirole response in reserpine-treated monkeys suggested that the impairments in delayed response and fine motor performance resulted from drug actions at D2 autoreceptors, while the improvement in delayed response performance, dyskinesias and ''hallucinatory-like'' behaviors resulted from actions at postsynaptic receptors. In aged monkeys, low doses of quinpirole continued to impair fine motor performance, but lost their ability to impair delayed response performance. The magnitude of cognitive improvement and the incidence of ''hallucinatory-like'' behaviors were also reduced in the aged animals, suggesting some loss of postsynaptic D2 receptor function, The pattern of results is consistent with the greater loss of DA from the PFC than from motor areas in aged monkey brain (Goldman-Rakic and Brown, 1981; Wenk et al., 1989), and indicates that DA depletion contributes significantly to age-related cognitive decline.

Effects of Naotan Pill on repair of neural cells and cognitive disorders in juvenile rats following hypoxia and ischemia

BACKGROUND: Hypoxia and ischemia induce neuronal damage, decreased neuronal numbers and synaptophysin levels, and deficits in learning and memory functions. Previous studies have shown that lycium barbarum polysaccharide, the most effective component of barbary wolfberry fruit, has protective effects on neural cells in hypoxia-ischemia. OBJECTIVE: To investigate the effects of Naotan Pill on glutamate-treated neural cells and on cognitive function in juvenile rats following hypoxia-ischemia. DESIGN, TIME AND SETTING: The randomized, controlled, in vivo study was performed at the Cell Laboratory of Lanzhou University, Lanzhou Institute of Modern Physics of Chinese Academy of Sciences, and Department of Traditional Chinese Medicine of Gansu Provincial Rehabilitation Center Hospital, China from December 2005 to August 2006. The cellular neurobiology, in vitro experiment was conducted at the Institute of Human Anatomy, Histology, Embryology and Neuroscience, School of Basic Medical Sciences, Lanzhou University, and Department of Traditional Chinese Medicine of Gansu Provincial Rehabilitation Center Hospital, China from March 2007 to January 2008. MATERIALS: Naotan Pill, composed of barbary wolfberry fruit, danshen root, grassleaf sweetflag rhizome, and glossy privet fruit, was prepared by Gansu Provincial Rehabilitation Center, China. Rabbit anti-synaptophysin, choline acetyl transferase polyclonal antibody, streptavidin-biotin complex kit and diaminobenzidine kit (Boster, Wuhan, China), as well as glutamate (Hualian, Shanghai, China) were used in this study. METHODS: Cortical neural cells were isolated from neonatal Wistar rats. Neural cell damage models were induced using glutamate, and administered Naotan Pill prior to and following damage. A total of 54 juvenile Wistar rats were equally and randomly assigned into model, Naotan Pill, and sham operation groups. The left common carotid artery was ligated, and then rat models of hypoxic-ischemic injury were assigned to the model and Naotan Pill groups. At 2 days following model induction, rats in the Naotan Pill group were administered Naotan Pill suspension for 21 days. In the model and sham operation groups, rats received an equal volume of saline. MAIN OUTCOME MEASURES: Neural cell morphology was observed using an inverted phase contrast microscope. Survival rate of neural cells was measured by MTT assay. Synaptophysin and choline acetyl transferase expression was observed in the hippocampal CA1 region of juvenile rats using immunohistochemistry. Cognitive function was tested by the Morris water maze. RESULTS: Pathological changes were detected in glutamate-treated neural cells. Neural cell morphology remained normal after Naotan Pill intervention. Absorbance and survival rate of neural cells were significantly greater following Naotan Pill intervention, compared to glutamate-treated neural cells (P < 0.05). Synaptophysin and choline acetyl transferase expression was lowest in the hippocampal CA1 region in the model group and highest in the sham operation group. Significant differences among groups were observed (P < 0.05). Escape latency and swimming distance were significantly longer in the model group compared to the Naotan Pill group (P < 0.05). CONCLUSION: Naotan Pill exhibited protective and repair effects on glutamate-treated neural cells. Naotan Pill upregulated synaptophysin and choline acetyl transferase expression in the hippocampus and improved cognitive function in rats following hypoxia-ischemia.

中老年人工作记忆与血管性疾病及危险因素的相关分析

Background With the increasing of an elderly population, the number of both vascular disease and cognitive function is increasing. Recent study shown that there is close relationship between vascular disease or vascular risk factors and cognitive function, but many questions are unclear. Objective The aim of this study was to explore the status of cognitive function of midlife and elderly subjects and to evaluate association between vascular risk factors and cognitive function. Methods We selected 330(female,128,male,202) retire navy veterans aged between 55 and 87 years old in Beijing. The average age was 67 years old. Data about vascular risk factors, including: history of cerebral infarction, cardiovascular disease, hypertension, diabetes mellitus, carotid lesions, cigarette smoking, drinking, obesity, and so on. Cognitive functions were assessed by neuropsychological tests: the four tasks of working memory. Results The univariate analysis indicated that there were significant differences of working memory scores in aging, gender, education level, the history of stroke, hypertension with midlife and elderly subjects. Moderate levels of alcohol intake maybe better for cognition. Conclusion vascular disease and some vascular risk factors may lead to cognitive function decline, thus, intervention methods should be carried out in middle and elderly people with vascular risk fact should be under special supervision, with at least annual neuropsychological evaluation. Key words: cognitive function; vascular risk factor， working memory.

2型糖尿病患者认知功能特点及相关影响因素研究

Objective: Type 2 diabetes patients’ performances of action memory , semantic memory and working memory and the related factors were explored. Methods: 60 Type 2 diabetes patients were compared with 60 age and gender and level of education matched non-diabetes controls. Mood were tested by SAS and SDS, MMSE was used to test the basic cognitive function, Trail Making Test A and B, Verbal fluency test, Go-No/Go test, and Stroop color-word test were used to investigate the executive function of Type 2 diabetes patients and normal controls (NC). Patients’ GLU, TG, TCH, HbA1c, insulin and Cp were tested and correlated with their action memory and working memory. Results: There was no difference between NC group and Type 2 diabetes patients in MMSE scores. There is depression and anxiety mood in Type 2 diabetes patients. Type 2 diabetes patients get lower score in action memory test. Comparing to NC group, Type 2 diabetes patients performed significantly worse in Trail Making Test A and B and verbal fluency test. In Stroop Test, NC group showed significant Stroop Effect and Repeated Distraction Promotion Effect and Negative Priming Effect. However, In Type 2 diabetes group, only the Stroop Effect appeared, but no Repeated Distraction Promotion Effect and Negative Priming Effect. There is no difference between Type 2 diabetes and NC in Stroop Effect. In Go-No/Go test, both of two groups showed significant Stroop Effect, however, there was no difference between them. And also there is no difference on error rate of all levels between them. The course of disease, GL, HbA1c, TG, TCH, INS and Cp affected action memory and working memory. Conclusion: Type 2 diabetes patients’ action memory, semantic memory and working memory were partially impaired. Controlling the levels of GLU, TG and TCH can delay these kinds of impairment.

RESERPINE IMPAIRS SPATIAL WORKING-MEMORY PERFORMANCE IN MONKEYS - REVERSAL BY THE ALPHA-2-ADRENERGIC AGONIST CLONIDINE

Repeated daily treatment with the catecholamine-depleting agent, reserpine, dramatically reduced performance on the delayed response task, a test of spatial working memory that depends upon the integrity of the prefrontal cortex. Delayed response performance fell from an average of 27.2/30 trials correct before reserpine treatment to an average of 20.4/30 trials correct after repeated reserpine administration. Injection of the alpha2-adrenergic agonist, clonidine (0.0001-0.05 mg/kg), to chronic reserpine-treated monkeys significantly restored performance on the delayed response task; performance after an optimal dose averaged 27.8/30 trials correct. Clonidine's beneficial effects on delayed response performance were longlasting; monkeys remained improved for more than 24 h after a single clonidine injection. The finding that clonidine is efficacious in reserpinized animals supports the hypothesis that alpha2-adrenergic agonists improve cognitive function through actions at postsynaptic, alpha2-adrenergic receptors on non-adrenergic cells. In contrast to the delayed response task, reserpine had little effect on performance of a visual discrimination task, a reference memory task which does not depend on the prefrontal cortex. These results emphasize the importance of postsynaptic alpha2-adrenergic mechanisms in the regulation of working memory,

Dose-dependent effects of the dopamine D1 receptor agonists A77636 or SKF81297 on spatial working memory in aged monkeys

With advancing age, monkeys develop deficits in spatial working memory resembling those induced by lesions of the prefrontal cortex (PFC). Aged monkeys also exhibit marked loss of dopamine from the PFC, a transmitter known to be important for proper PFC cognitive function. Previous results suggest that D1 agonist treatment can improve spatial working memory abilities in aged monkeys. However, this research was limited by the use of drugs with either partial agonist actions or significant D2 receptor actions. In our study, the selective dopamine D1 receptor full agonists A77636 and SKF81297 were examined in aged monkeys for effects on the working memory functions of the PFC. Both compounds produced a significant, dose-related effect on delayed response performance without evidence of side effects: low doses improved performance although higher doses impaired or had no effect on performance. Both the improvement and impairment in performance were reversed by pretreatment with the D1 receptor antagonist, SCH23390. These findings are consistent with previous results demonstrating that there is a narrow range of D1 receptor stimulation for optimal PFC cognitive function, and suggest that very low doses of D1 receptor agonists may have cognitive-enhancing actions in the elderly.

Extract of Ginkgo biloba leaves reverses yohimbine-induced spatial working memory deficit in rats

Extract of Ginkgo biloba is used to alleviate age-related decline in cognitive function, which may be associated with the loss of catecholamines in the prefrontal cortex. The purpose of this study was to verify whether alpha-2 adrenergic activity is involved in the facilitative effects of extract of Ginkgo biloba on prefrontal cognitive function. Male Wistar rats were trained to reach criterion in the delayed alternation task (0, 25, and 50-s delay intervals). A pilot study found that 3 or 4 mg/kg of yohimbine (intraperitoneal) reduced the choice accuracy of the delayed alternation task in a dose and delay-dependent manner, without influencing motor ability or perseverative behaviour. Acute oral pre-treatment with doses of 50, 100, or 200 mg/kg (but not 25 mg/kg) of extract of Ginkgo biloba prevented the reduction in choice accuracy induced by 4 mg/kg yohimbine. These data suggest that the prefrontal cognition-enhancing effects of extract of Ginkgo biloba are related to its actions on alpha-2-adrenoceptors.

Disconnection of the hippocampal-prefrontal cortical circuits impairs spatial working memory performance in rats

There is a unidirectional, ipsilateral and monosynaptic projection from the hippocampus to the prefrontal cortex. The cognitive function of hippocampal-prefrontal cortical circuit is not well established. In this paper, we use muscimol treated rats to inv

The in vivo synaptic plasticity mechanism of EGb 761-induced enhancement of spatial learning and memory in aged rats

1 It has not been uniform to date that the Ginkgo biloba extracts enhance cognitive function in aged animals, and the mechanisms of action remain difficult to elucidate. In this study, the Morris water maze task and electrophysiological methods were used

Exact result for the Green's function of a two-dimensional interacting electron gas with spin-orbit interaction in a perpendicular magnetic field

An exact property is established for the Green's function of a uniform two-dimensional interacting electron gas in a perpendicular magnetic field with spin-orbit interaction. It is shown that the spin-diagonal Green's function is exactly diagonal in the Landau level index even in the presence of electron-electron interactions. For the Green's function with different spin indexes, only that with adjacent Landau level indexes is non-zero. This exact result should be helpful in calculating the Green's function approximately.

1.老年猴视皮层神经元对图形对比度的反应及其潜伏期特征; 2. 极低频磁场对脑功能的影响及眶额叶认知功能的研究

1.老年猴视皮层神经元对图形对比度的反应及潜伏期特征: 在正常衰老过程中人类的视觉功能受到严重影响，例如空间和时间对比度敏感性下降以及信息处理时间的延长。虽然部分视觉功能的退化与眼睛的光学系统老化有关，但是它并不能解释所有视觉功能的下降。此外，我们以前的研究和别人的研究结果都表明衰老过程中视觉中枢系统功能的改变可能是视觉功能下降的主要原因。因此，利用单位放电记录技术（single-unit recording technique），我们比较了年轻猕猴和老年猕猴的初级视觉皮层（primary visual cortex，又称V1）神经元对比度反应之间的差异，以及V1和内侧颞叶（medial temporal cortex，MT）视觉区神经元反应潜伏期及其变异性之间的差异。结果显示，与年轻猴相比，老年猴V1区神经元对比度敏感性降低，同时伴随着神经元活动信噪比下降；老年猴V1区和MT区神经元反应潜伏期及其变异性显著增加。然而，两个年龄组MT区神经元平均潜伏期之间差异小于V1区神经元平均潜伏期之间的差异，说明MT区神经元能够自我调整老化带来的影响。另外，两个年龄组V1区神经元潜伏期和变异性都具有正相关关系，但是MT区神经元则没有这种相关性。这些结果表明，在老化过程中皮层神经元的对比度和潜伏期反应特性发生了改变。我们推测这种改变可能与视觉皮层内抑制系统功能的降低有关，但是具体的分子机制和神经环路还不清楚。总之，本实验的研究结果为更好的理解老年人在视觉信息处理中时间和空间对比度敏感性及处理速度下降提供了新线索。2.极低频磁场对脑功能的影响及眶额叶认知功能的研究: 实验目的：（1）研究极低频磁场(20 Hz, 14 mT)照射对长期吗啡处理引起的大鼠背侧海马神经元多巴胺D2密度降低的影响；（2）小鼠青春期长期极低频磁场(50 Hz, 2 mT)照射对空间学习记忆的影响；（3）初步探讨了眶额叶在大鼠新异性探索行为中的作用。实验1，我们用免疫组化的方法检测了大鼠背侧海马神经元多巴胺D2受体密度的变化。结果显示，在长期吗啡处理后戒断早期背侧海马神经元多巴胺D2受体密度相对于对照组减少，磁场和吗啡共同作用会强化这种适应，但是这种变化很快恢复正常。这些结果表明长期吗啡处理会引起海马多巴胺系统产生适应；磁场强化了长期吗啡处理对背侧海马多巴胺系统的影响,这为我们先前发现磁场照射延缓了大鼠条件位置偏好消退的研究结果提供了一个内在神经基础。实验2，我们分别用Y-迷宫（two-trial Y-maze）和Morris水迷宫两种行为装置研究了青春期早期磁场暴露对小鼠短时空间识别记忆和长时空间参考记忆的影响。结果显示，磁场暴露并没有影响小鼠Y-迷宫作业，但是提高了水迷宫任务的学习以及记忆保持。这些结果表明磁场对空间记忆的影响是任务依赖性的。实验3，我们用旷场和Y-迷宫两种行为装置研究了眶额叶电损伤对大鼠新异性探索行为的影响。结果显示，眶额叶受损并没有影响大鼠的神经运动能力，但是降低了大鼠在旷场中的行走距离和直立次数以及降低了在Y-迷宫新异臂中的探索时间和穿梭次数。这些结果表明，眶额叶的完整性对大鼠探索新异环境行为是必要的，这可能与眶额叶参与记忆或行为决策功能有关。

维持性血液透析患者认知功能特点研究

Objective: To study the episodic memory, semantic memory, cognitive planning ability and inhibition ability in MHD patients. Method: Neuropsychological research methods such as Action memory of verb-object phrase, Trail Making Test (A and B), Verbal Fluency Test, Go-No/Go test and Stroop Color Naming Task were used to investigate Episodic Memory 、Semantic Memory、Executive Function of 40 MHD and 40 NC. Self-Rating Anxiety Scale (SAS), Self-Rating Depression Scale(SDS), Social Support Scale, Life Satisfaction Scale, and biochemical examination were applied and their relationships with cognitive function were analized. The mean age and education level of MHD group and NC group have no significant difference. Result: 1.Action memory of verb-object phrase differed significantly between MHD group and NC group. 2.Two tests of Verbal Fluency differed significantly between MHD group and NC group. 3.Trail Making Test A, Trail Making Test B, the baseline condition of Go-No/Go Test and Stroop Color Naming Test differed significantly between MHD group and NC group. 4.There is no significant difference between MHD group and NC group on the correct rate of No/Go Test and the baseline condition. Both groups showed Stroop Effect in Go-No/Go test, but MHD group performed significantly worse. 5.In Stroop Color Naming Task Test, NC group showed Stroop Effect, significant Repeated Distraction Promotion Effect and significant Negative Priming Effect,while MHD group showed only Stroop Effect and no Repeated Distraction Promotion Effect and no Negative Priming Effect. There is significant difference in Stroop Effect between MHD group and NC group. Conclusion: 1.Comparing with NC group, episodic memory, semantic memory, cognitive planning ability, and inhibition ability of MHD group were impaired significantly. 2.The pathological aging of Executive Function in MHD group showed: executive Function should be a unitary system. 3.Cognitive impairment is negatively correlated with serum creatinine, blood pressure and anxiety score in MHD patients; and is related with hemoglobin, hematocrit, social support and life satisfaction. Keyword: maintenance hemodialysis, episodic memory, semantic memory, cognitive planning, inhibition ability.