补骨脂素干预大鼠成骨细胞骨保护素/核因子κB受体激活因子配体mRNA的表达

背景：补骨脂素有促进大鼠成骨细胞增殖与分化的作用,但其作用机制尚不明确。 目的：探讨补骨脂素对大鼠成骨细胞中骨保护素(osteoporogeterin, OPG)和核因子κB受体激活因子配体(receptor activator nuclear factor kappa b ligand, RANKL) mRNA表达的影响。 方法：取第3代生长状况良好的出生24 h内的SD大鼠成骨细胞,补骨脂素组加入1×10-7 mol/L的补骨脂素,雌二醇组加入1×10-7 mol/L的雌二醇,对照组正常培养。给药72 h后提取细胞总RNA,RT-PCR方法分析细胞OPG/RANKL mRNA的表达。 结果与结论：与对照组比较,补骨脂素组和雌二醇组OPG mRNA的表达均明显增加(P < 0.05),而RANKL mRNA的表达明显下降(P < 0.05),但补骨脂素组成骨细胞OPG mRNA的表达较雌二醇组弱(P < 0.05),VOPG/VRANKL比值也较雌二醇组小(P < 0.05)。说明补骨脂素可能通过增加成骨细胞OPG的表达,抑制RANKL的表达来抑制破骨细胞的分化和成熟,从而抑制骨吸收,达到防治骨质疏松症的目的,但作用不如雌二醇明显。     

异欧前胡素对大鼠成骨细胞增殖与分化的影响

背景：植物雌激素有防治绝经后妇女骨质疏松的作用已得到公认,其作用机制之一是促进了成骨细胞的增殖与分化。 目的：观察香豆素类植物雌激素异欧前胡素体外对大鼠成骨细胞增殖与分化的影响。 设计、时间及地点：对比观察实验,于2006-09/2007-12在河北医科大学药学院完成。 材料：出生24 h内SD大鼠,异欧前胡素为中国药品生物制品检定所产品。 方法：采用改良组织块法分离培养新生大鼠颅骨成骨细胞,将异欧前胡素以1×10-5~ 1×10-9 mol/L浓度加入细胞培养体系,以未加入药物为空白对照组。作用24,48,72 h后,以MTT法检测成骨细胞的增殖情况,以对硝基苯二钠基质动力学法测定细胞内碱性磷酸酶的活性,以改良Lowry法测总蛋白水平。 主要观察指标：成骨细胞的增殖率和碱性磷酸酶活性。 结果：大鼠成骨细胞在不同浓度异欧前胡素中培养24 h,未观察到促增殖作用。培养48 h后,开始出现促增殖作用增强,有效浓度为1×10-9~1×10-8 mol/L。培养72 h后,继续有促增殖作用,有效浓度为1×10-8~1×10-7mol/L。大鼠成骨细胞在不同浓度异欧前胡素存在下培养48 h,未观察到促分化作用,培养72 h后,开始出现促分化作用,有效浓度为1×10-6~1× 10-5 mol/L,其他浓度作用不明显。 结论：异欧前胡素体外能促进大鼠成骨细胞的增殖与分化。     

不同浓度硝酸银负载珊瑚羟基磷灰石的细胞毒性实验***☆

背景：硝酸银负载珊瑚羟基磷灰石是一种具有抗菌性能的新型植骨材料,在国外已有大量的研究报道,但国内对其研究仅处于起步阶段。 目的：制备载银珊瑚羟基磷灰石人工骨材料并观察其对L929细胞的毒性特征。 设计、时间及地点：细胞学体外对比观察实验,于2008-10/11在解放军广州军区广州总医院中心实验室完成。 材料：将自制的珊瑚羟基磷灰石浸泡于1×10-2,1×10-3,5×10-4,1×10-4,5×10-5,1×10-5 mol/L硝酸银溶液中制得不同浓度的载银人工骨材料。按1.25 cm2材料表面积/1 mL培养基比例加入培养基,37 ℃下浸提24 h。 方法：将材料浸提培养基与L929细胞共培养48 h,运用MTT法测得细胞相对增殖度,用倒置相差显微镜观察细胞形态,结合中华人民共和国国家标准：GB/T16175-2008中的相关标准进行分析。 主要观察指标：L929细胞在不同材料上的增殖情况及细胞的形态学变化。 结果：MTT法检测结果表明硝酸银浓度≤5×10-5 mol/L时,材料对L929细胞生长无明显抑制作用;材料浸提液即使在高浸提比100%下,浸提产物也无细胞毒性作用。镜下观察发现5×10-5 mol/L组及1×10-5 mol/L组细胞形态正常,贴壁生长良好。 结论：低浓度的载银珊瑚羟基磷灰石对体外培养的L929细胞的增殖、生长无明显毒性作用。 关键词：珊瑚羟基磷灰石;细胞毒性;MTT比色法;抗感染骨替代材料;银离子     

唑来膦酸对大鼠成骨细胞增殖、分化和护骨素/核因子κB受体激动剂配体mRNA表达的影响☆

背景：在应用于预防和治疗假体周围骨溶解时，唑来膦酸可在发挥抑制破骨细胞作用的同时而不影响成骨功能。 目的：观察唑来膦酸对大鼠成骨细胞功能的影响，验证其应用于人工关节置换后假体周围骨溶解的可行性。 设计、时间及地点：观察对照实验，于2007-03/2008-03在华北煤炭医学院中心实验室完成。 材料：新生24 h内SD大鼠，唑来膦酸标准品。 方法：体外培养新生大鼠颅盖骨来源的成骨细胞，应用不同浓度唑来膦酸进行干预，实验组细胞培养基中唑来膦酸终浓度为10-4~10-11 mol/L，对照组不加药, 只加等量细胞悬液和培养基。唑来膦酸干预后分别于第1，2，3，5，7天采用四甲基偶氮唑盐比色法测定吸光度，检测唑来膦酸对成骨细胞增殖的影响；第3天和第5天分别采用荧光定量聚合酶链反应测护骨素和核因子κB受体激动剂配体mRNA表达，以硝基苯基质动力学法测定各组细胞碱性磷酸酶活性。 主要观察指标：成骨细胞增殖情况和碱性磷酸酶活性，护骨素、核因子κB受体激动剂配体mRNA表达水平。 结果：唑来膦酸浓度≥10-5 mol/L时抑制成骨细胞增殖，10-6~10-11 mol/L则不影响细胞增殖。唑来膦酸浓度为10-7 ~10-11 mol/L时碱性磷酸酶活性无变化，而成骨细胞的护骨素mRNA表达增加，核因子κB受体激动剂配体mRNA的表达下降。 结论：唑来膦酸在低浓度时不影响成骨细胞增殖及分化，通过降低成骨细胞核因子κB受体激动剂配体/护骨素的比率而抑制破骨细胞分化，可应用于人工关节置换后假体周围骨溶解。     

唑来膦酸对大鼠成骨细胞护骨素及肿瘤坏死因子α mRNA表达的影响

背景：唑来膦酸属于第3代双瞵酸盐类药物,在临床上被广泛应用于治疗骨吸收增加类疾病,其对破骨细胞的作用及影响已取得了共识,而对于成骨细胞的影响尚有争议。 目的：观察唑来膦酸对大鼠成骨细胞增殖、分化和护骨素及肿瘤坏死因子α mRNA表达的影响。 方法：体外培养新生24 h内SD大鼠颅盖骨来源的成骨细胞,用10-5~10-9 mol/L唑来膦酸进行干预,分别于干预后第3,5,7天采用MTT法来测吸光度值,以检测唑来膦酸对大鼠成骨细胞增殖的影响;硝基苯基质动力学法和RT-PCR在干预后72 h,测量细胞碱性磷酸酶活性和护骨素及肿瘤坏死因子α mRNA的表达。 结果与结论：较高浓度(10-5 mol/L)的唑来膦酸明显抑制成骨细胞增殖, 10-7~10-9 mol/L唑来膦酸不影响成骨细胞的分化。10-5~10-9 mol/L唑来膦酸能使成骨细胞护骨素mRNA表达增加,肿瘤坏死因子α mRNA表达下降。说明唑来膦酸在低浓度时不影响成骨细胞的增殖及分化,其可通过调节成骨细胞护骨素、肿瘤坏死因子α mRNA的表达,来发挥其抗骨吸收的作用。     

假体磨损颗粒钴铬离子影响成骨细胞增殖及RANKL、骨保护素的表达

背景：金属-金属假体置入体内后可以发生腐蚀或磨损,释放镍、钴、铬、钛等金属离子,诱导局部炎性因子的释放。 目的：观察Co2+、Cr3+对小鼠成骨细胞增殖的影响,以及成骨细胞暴露在Co2+ 、Cr3+条件下RANKL、骨保护素基因的表达。 方法：体外培养成骨细胞,实验分两组,对照组给予生理盐水,离子组给予钴、铬离子干预。 结果与结论：显微镜直接计数法显示干预后1~6 d对照组随时间推移细胞数目显著增加,离子组则增加不明显。共培养24,48 h后,RT-PCR结果显示离子组RANKL、骨保护素基因表达较对照组均增加,以RANKL增加更显著(P < 0.05),RANKL/OPG mRNA的比率也明显增加。提示金属离子对成骨细胞的增殖有显著抑制作用,且可刺激成骨细胞RANKL、骨保护素mRNA的表达。     

雌激素对成骨细胞血管内皮生长因子表达的影响

背景：在众多的血管生长因子中,血管内皮生长因子是最有力的血管生成因子,它具有促进血管内皮细胞分裂、增殖,细胞质钙化聚集并能诱导血管生成等作用,且对于成骨细胞、破骨细胞的增殖、分化及功能活性具有重要调节作用。 目的：观察雌激素对成骨细胞上血管内皮生长因子表达的影响。 方法：取新生48 h大鼠颅盖骨,酶解消化得到成骨细胞,并进行体外培养。采用RT-PCR法测定10-10,10-8,10-6,10-4 mol/L 17β-雌二醇作用成骨细胞后血管内皮生长因子mRNA的表达。 结果与结论：RT-PCR检测结果显示,血管内皮生长因子mRNA在原代成骨细胞内稳定表达;半定量分析证实,不同浓度17β-雌二醇组间血管内皮生长因子mRNA的表达量呈现一定规律,随着17β-雌二醇浓度的升高,血管内皮生长因子mRNA的表达量逐渐增加,10-6 mol/L左右时,血管内皮生长因子mRNA的表达量最高,超过此剂量,血管内皮生长因子的表达下降。提示雌激素促进成骨细胞上血管内皮生长因子的表达上调,并存在剂量依赖性。     

聚甲基丙烯酸甲酯颗粒对人滑膜细胞核因子κB受体激动剂配体/骨保护蛋白表达的影响

背景：多种因子可以影响骨保护蛋白/核因子κB受体激动剂配体/核因子κB受体激动剂(Osteoprotegerin / receptor activator of nuclear factor-kappaB/ligand of receptor-activator of nuclear factor-kappaB,OPG/RANKL/RANK)系统的表达影响骨代谢,那么松动假体周围的骨水泥颗粒是否也通过影响其代谢参与假体松动过程？ 目的：观察聚甲基丙烯酸甲酯颗粒对人滑膜细胞RANKL/OPG表达的影响。 方法：体外培养人滑膜细胞,在滑膜细胞培养体系中分别加入质量浓度为0(空白对照),0.2,1,10 g/L的聚甲基丙烯酸甲酯骨水泥颗粒,采用荧光定量PCR检测上述各浓度PMMA颗粒作用不同时间后滑膜细胞内RANKL、OPG的表达量及其比值。 结果与结论：与空白对照组相比,各实验组滑膜细胞内RANKL、OPG的表达量均有下降;随着与聚甲基丙烯酸甲酯颗粒共培养时间延长,各实验组滑膜细胞内RANKL、OPG表达均有下降趋势,而RANKL/OPG表达比值无明显变化。说明聚甲基丙烯酸甲酯颗粒在对滑膜细胞产生生物学反应时并不干扰假体周围骨代谢的动态平衡,并未直接参与人工关节假体的无菌性松动。     

降钙素对大鼠颅骨成骨细胞调控作用的体外观察

背景：降钙素是强有力的破骨细胞抑制剂,可直接、快速而广泛地抑制骨吸收,近年来有学者报道其有促进骨折愈合的功能,推测降钙素对成骨细胞也有直接的作用。 目的：体外观察降钙素对大鼠成骨细胞的药理作用,分析其是否通过细胞外信号调节激酶信号通路产生。 设计、时间及地点：分组对照观察,于2007-06/2008-05在南京医科大学药理学实验室完成。 材料：选用新生SD大鼠,分离培养颅骨成骨细胞。 方法：取第2代成骨细胞进行分组实验：0.01 μg/L降钙素组、0.1 μg/L降钙素组、1 μg/L降钙素组分别加入不同剂量降钙素进行干预,U0126＋1 μg/L降钙素组提前30 min加入100 μmol/L的细胞外信号调节激酶抑制剂U0126,并设空白对照组。 主要观察指标：通过四甲基偶氮唑盐比色法、对硝基苯磷酸盐法和钙结节茜素红染色法分别检测降钙素对成骨细胞的增殖、分化、矿化能力的影响;应用反转录-聚合酶链反应技术检测促骨形成关键基因核心结合因子a1 mRNA的表达情况;应用Western Blotting技术检测磷酸化细胞外信号调节激酶1/2蛋白表达情况。 结果：加入0.01,0.1,1 μg/L的降钙素干预后,成骨细胞的增殖、分化、矿化能力以及核心结合因子a1 mRNA的表达均随降钙素剂量增加而增强,且0.1,1 μg/L组与空白对照组比较差异具有显著意义（P < 0.05~0.01）;此外降钙素促进了磷酸化细胞外信号调节激酶1/2蛋白的表达(P < 0.05)。U0126可阻断以上所有效应（P < 0.05~0.01）。 结论：降钙素可能通过细胞外信号调节激酶信号通路调控促骨形成关键基因核心结合因子a1 mRNA的表达,进而促进成骨细胞的增殖、分化、矿化能力。     

成骨生长肽对骨髓基质细胞增殖和向成骨方向分化的影响

背景: 成骨生长肽是新近发现的一种具有促进成骨作用的生长因子,其对骨髓基质细胞增殖分化作用受到越来越多的关注。 目的：验证成骨生长肽在体外对骨髓基质细胞增殖和成骨向分化的影响。 设计、时间及地点：随机对照实验,于2006-02/06在教育部口腔生物医学工程重点实验室完成。 材料：6周龄雄性SD大鼠用于骨髓基质细胞提取;成骨生长肽为Sigma公司产品。 方法：体外培养骨髓基质细胞,加入含不同浓度(10-11,10-10,10-9,10-8,10-7 mol/L)成骨生长肽及小牛血清的α-MEM培养基培养,以单纯含小牛血清的α-MEM培养基培养作对照。 主要观察指标：①骨髓基质细胞的鉴定。②用四甲基偶氮唑盐法检测细胞增殖情况。③酶联免疫法检测细胞内碱性磷酸酶的表达。④反转录-聚合酶链反应法检测细胞内核心结合因子1mRNA的表达。 结果：成骨生长肽对骨髓基质细胞增殖有促进作用(P < 0.05),最大效应浓度为 10-9 mol/L;能增加细胞内碱性磷酸酶活性(P < 0.05),最大效应浓度为10-8 mol/L;可诱导核心结合因子1mRNA的表达(P < 0.05)。 结论：成骨生长肽可促进骨髓基质细胞的增殖,并通过诱导其内核心结合因子1mRNA的表达促进其成骨向的分化。     

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.