IFN-γ和热干预对U251胶质瘤细胞的MHC-Ⅰ类分子和HSP70表达的影响

目的观察重组人干扰素-γ（IFN-γ）和热干预对人多形胶质母细胞瘤（GBM）U251细胞胞膜主要组织相容性抗原复合体-Ⅰ（MHC-Ⅰ）类分子和热休克蛋白70（HSP70）分子表达的影响。方法将U251细胞分为3个干预组：IFN干预组（IFN-γ500U/ml诱导48h）,热干预组（43℃热休克2h）,联合干预组（IFN-γ500U/ml诱导48h＋43℃热休克2h）,分别行相应诱导干预。流式细胞仪（FCM）检测不同干预因素处理后的U251细胞胞膜MHC-Ⅰ类分子和HSP70表达情况。结果IFN干预组、热干预组、联合干预组U251细胞胞膜MHC-Ⅰ及HSP70表达率分别为（89.92±3.45）%和（5.11±1.89）%、（78.89±2.02）%和（42.18±3.85）%、（96.00±2.63）%和（53.61±4.24）%。结论双因素（IFN-γ500U/ml诱导48h＋43℃热休克2h）联合干预是U251细胞胞膜MHC-Ⅰ类分子和HSP70双高表达的最佳体外诱导方案。     

高免疫原性胶质瘤细胞疫苗体外诱导Th-1漂移研究

目的 观察高免疫原性即膜型热休克蛋白70(HSP70)及主要组织相容性抗原复合体Ⅰ类分子(MHC-Ⅰ)双高表达人多形性胶质母细胞瘤U251(GBM U251)细胞疫苗在体外诱导Th-1漂移现象.方法 GBM U251分别以500 U/mL IFN-γ诱导48 h、43℃热休克2 h、500 U/mLIFN-γ诱导48 h+43℃热休克2 h诱导其MHC-Ⅰ类分子、膜型HSP70高表达,随后经丝裂霉素(MMC)灭活制成细胞疫苗.体外刺激健康捐献者外周血单个核细胞(PBMCs)作为效应细胞,进行肿瘤特异性杀伤试验;FCM检测疫苗刺激前后PBMCs CD4<'+>、CD8<'+>T淋巴细胞比例变化;ELISA法检测效应细胞攻击靶细胞后的IFN-γ、IL-2的分泌情况.结果 体外刺激后,HSP70和MHC-Ⅰ类分子双高表达的U251细胞疫苗CD4<'+>、CD8<'+>比例相对于MHC-Ⅰ类分子单高表达或HSP70分子单高表达细胞疫苗组明显增加,体外IFN-γ和IL-2分泌量亦增加,差异均有统计学意义(p<0.05).结论高免疫原性即HSP70和MHC-Ⅰ类分子双高表达U251细胞疫苗体外诱导产生Th-1漂移现象,推测是其体外抗瘤作用的重要机制之一.     

HSP70调节ATO诱导胶质瘤细胞死亡的实验研究

目的通过调节热休克蛋白70(HSP70)的表达,观察HSP70在三氧化二砷(ATO)诱导胶质瘤细胞U251MG死亡中的作用。方法台盼蓝染色用来评价细胞的活性,免疫印迹分析HSP70和Caspase-3表达情况,进一步使用热休克蛋白抑制剂KNK437和热激的方法调节HSP70的水平,观察ATO在不同HSP70水平对U251MG死亡的影响。结果 ATO诱导U251MG细胞的死亡,伴有HSP70的表达;应用KNK437明显增加ATO诱导U251MG细胞的死亡和凋亡蛋白Caspase-3表达;热激明显抑制ATO诱导U251MG细胞的死亡。结论 HSP70在ATO诱导胶质瘤细胞U251MG死亡中起保护作用,KNK437可能在ATO治疗胶质瘤中起协同作用。     

热休克蛋白减轻1-甲基-4-苯基-吡啶离子引起的线粒体功能障碍和氧化应激

目的观察热休克蛋白(HSP)对1-甲基-4-苯基-吡啶离子(MPP )引起的线粒体功能障碍和氧化应激的保护作用。方法采用免疫印迹法观察热休克诱导HSP的表达以及转染的HDJ-1基因在细胞内的过表达。通过5,5′,6,6′-四氯-1,1′,3,3′-四乙基苯丙咪唑羰花青碘化物(JC-1)和2′,7′-二氯荧光黄双乙酸盐(DCFH-DA)流式细胞术及荧光显微镜观察MPP 对细胞线粒体膜电势和活性氧族(ROS)的影响以及HSP的保护作用。结果热休克后4h即有Hsp70(7.37±1.17)和HDJ-1(2.32±0.37)增加,并至少持续到72h;同样,转染24h后HDJ-1基因在细胞内过表达(1.26±0·06),并至少持续到72h。MPP 能引起线粒体膜电势降低(60.77±3.68),同时细胞内ROS上升(483.18±16.98)。热休克和HDJ-1基因过表达不仅能维持线粒体膜电势(热休克组68.32±3.42,转HDJ-1组66.13±3.31),而且还能抑制ROS的产生(热休克组449.45±18.80,转HDJ-1组470.56±23.53),其中热休克的作用更强。结论HSP通过保护线粒体功能、减少氧化应激来减轻MPP 毒性,从而发挥保护细胞的作用。     

IL-32在神经胶质瘤中的表达及调节机制研究

目的 探讨白细胞介素(IL)-32在神经胶质瘤中的表达及调节机制. 方法 RT-PCR、Western blotting检测体外培养3d的神经胶质瘤细胞株CHG-5、U251 IL-32 mRNA和蛋白的表达;应用10 ng/mL IL-1β、IL-4、IL-6、IL-10、IL-17、肿瘤坏死因子(TNF)-α、TNF-β、干扰素(IFN)-γ作用U251细胞24 h后RT-PCR检测细胞IL-32 mRNA表达的变化;RT-PCR检测不同浓度IL-1β、TNF-α、IFN-γ作用U251细胞不同时间后IL-32 mRNA表达的变化. 结果 U251细胞IL-32 mRNA表达水平高于CHG-5细胞,IL-32蛋白表达水平(0.95±0.42)高于CHG-5细胞(0.28±0.13),差异均有统计学意义(P＜0.05); IL-1β、TNF-α、IFN-γ作用U251细胞24 h后IL-32 mRNA表达量增加,差异有统计学意义(P＜0.05),且IL-32 mRNA的表达量对IL-1β、TNF-α、IFN-γ刺激的浓度和时间有依赖性. 结论 IL-32在神经胶质瘤中高表达,IL-1β、TNF-α、IFN-γ对IL-32 mRNA的表达具有调节作用,且存在时间与剂量依赖性.     

热休克蛋白70研究进展

热休克蛋白(HSP)是各种有机体在遭受应激刺激后诱导产生的一组应激蛋白。HSP70是HSP中最保守和最主要的一类,在细胞应激后生成最为显著,对细胞损伤具有保护作用,与缺血脑损伤等多种神经系统疾病关系密切。本文综述HSP70的结构、表达与调控、生物学功能及其与神经系统疾病的关系。     

大鼠局灶缺血预处理诱导的脑缺血耐受中热休克蛋白70的表达及其意义

目的　研究局灶脑缺血预处理对热休克蛋白 70 (HSP70 )表达和脑缺血耐受的影响。方法　SD大鼠随机分为 3组 :预缺血组、假手术组及对照组 ,前两组分别在 2小时大脑中动脉缺血 (MCAO)前 3天给予10分钟的预缺血或假手术 ,MCAO后 2 4小时处死 ,对照组给予两次相隔 3天的假手术 ,比较各组梗死体积及HSP70的表达。结果　预缺血组梗死体积较假手术组减少 5 2 5 4 % (P <0 0 1) ,HSP70表达高于假手术组及对照组 (P <0 0 1)。结论　 10分钟大脑中动脉预缺血可有效诱导缺血耐受 ,增加HSP70表达。HSP70表达上调可能是局灶性脑缺血耐受产生的分子机制之一     

替普瑞酮诱导阿尔茨海默病大鼠海马热休克蛋白70表达及抗凋亡机制

目的研究替普瑞酮(Geranylgeranylacetone,GGA)诱导阿尔茨海默病(Alzheimer's disease,AD)模型大鼠海马热休克蛋白70(heat shock protein 70,HSP70)的表达及其抗细胞凋亡机制。方法 36只SD大鼠随机分为对照组、模型组与GGA组(n=12)。双侧海马注射β淀粉样蛋白1-42(β-amyloid1-42,Aβ1-42)建立AD大鼠模型。GGA组大鼠予GGA800mg/(kg·d)灌胃,持续21d。Y迷宫测试大鼠行为学变化;HSP70-TUNEL免疫荧光双标染色观察大鼠海马细胞凋亡与HSP70表达变化、分布情况;Western-blot测定大鼠海马HSP70及凋亡相关因子caspase-9、cytochrome C、FADD、caspase-8蛋白表达。结果术后模型组大鼠获得记忆所需电击次数(14d:21.1±5.6;21d:27.4±5.2),对照组(14d:15.9±3.8;21d:14.7±3.9),GGA组(14d:17.3±2.7;21d:20.8±4.6)。术后21d海马HSP70表达:模型组(0.22±0.04),对照组(0.43±0.07),GGA组:(0.71±0.15);凋亡细胞:模型组(38.4±6.8),对照组(9.7±3.8);GGA组(16.7±4.3);caspase-9表达:模型组(0.69±0.09),对照组(0.14±0.04),GGA组(0.23±0.06)。FADD表达GGA组与模型组无统计学差异。结论 GGA诱导AD大鼠海马HSP70表达上调,可能通过抑制线粒体凋亡途径的激活及部分死亡受体凋亡途径的激活而发挥抗凋亡作用,减轻神经元损伤,从而改善AD大鼠的学习记忆能力。     

热休克蛋白70在阿尔茨海默病中作用的研究进展

目前的研究发现阿尔茨海默病(AD)患者脑中存在热休克蛋白70(HSP70)表达变化.HSP是机体在应激条件下产生的一组蛋白质,发挥分子伴侣作用而提高细胞对应激原的耐受性,维持细胞的正常生理功能.研究认为HSP70与AD的病理进程有关联, HSP70在抑制Aβ异常聚集及促进Aβ清除、抑制tau蛋白异常磷酸化及TNFs形成、阻断细胞凋亡途径、抗氧化应激等方面均发挥了重要作用,从而减轻AD多种因素造成的神经元损伤,本文就近年来HSP70在AD中作用的研究进展作一综述.     

一氧化氮对海人酸诱导大鼠癫痫发作脑组织海马结构HSP70表达的影响及意义

目的：研究海人酸诱导大鼠癫痫发作脑组织中一氧化氮（NO）与热休克蛋白70（HSP70）表达的关系。方法：利用免疫印迹分析法（westem blots)观察海人酸诱导大鼠癫痫发作及作用N－硝基－左旋精氨酸（LNNA）干预后脑组织海马结构HSP70的表达。结果：在癫痫发作过程中,预先用LNNA后,HSP70表达量较同一时间未干预的明显低。结论：在癫痫发作过程中,NO对HSP70表达有明显影响。     

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.