Distribution of nitric oxide synthase, nerve growth factor receptor and interstitial cells of Cajal in hirschsprung's disease and its significance

Objective To investigate the distribution of nitric oxide synthase (NOS), nerve growth factor receptor (NGFR) and interstitial cells of Caial (ICCs) in Hirschsprung's disease (HD). Methods The distribution of NOS,NGFR and ICCs was studied by using NADPH diaphoruse histochemistry, immanohistochemistry with a monoclonal antibody to human NGFR and the specific polycloual antibody against c-kit in 8 normal controls and 10 cases of HD.Results NOS and NGFR were abundantly present in the myenteric plexus and in the nerve fibers of musculature. ICCs were intensively distributed in the surface of circular musculature and around the myenteric plexus to form a network in normal control colon. In contrast, NOS and NGFR were scarce or absent in the myenteric plexus and in the nerve fibers of musculature, while the hypertrophic nerve trunks were NGFR positive, ICCs were scarcely distributed and the network was disrupted in the aganglionic colon in HD. Conclusion These findings suggest the Involvement of NOS,NGFR and ICCs in the patbophysiology of HD.     

尼古丁对巨噬细胞肝X受体α表达及胆固醇外流的影响

目的通过研究尼古丁对巨噬细胞的肝X受体α(LXRα)及其下游的基因表达和胆固醇外流的影响,探讨尼古丁对LXR信号系统的作用。方法分离人外周血单核细胞,并转化为巨噬细胞。在尼古丁的作用下,观察巨噬细胞的aopA-Ⅰ介导的胆固醇外流的变化和LXR及其下游一些基因mRNA表达。结果尼古丁显著影响巨噬细胞中一些涉及胆固醇代谢及炎症反应的基因表达,同时降低aopA-Ⅰ介导的胆固醇外流。结论尼古丁影响巨噬细胞LXR信号途径,促进泡沫细胞形成,使动脉粥样硬化发生发展。     

维生素E对糖尿病大鼠高级糖基化终末产物及其受体和肾功的影响

目的观察维生素E对糖尿病大鼠高级糖基化终末产物(AGEs)及其受体(RAGE)和肾功能的影响。方法利用链脲佐菌素腹腔注射法诱导建立糖尿病大鼠模型。将实验用SD大鼠随机分为正常对照组(NC组)、糖尿病组(DM组)和维生素E治疗组(VE组),治疗8周。观察大鼠的一般状况,监测血糖(BGL)、糖化血红蛋白(HbA1C)、尿素氮(BUN)、血肌酐(Scr)、内生肌酐清除率(Ccr)、尿白蛋白排泄率(UAER)、肾重/体重比值;应用荧光显微镜观察肾脏组织AGEs,免疫组化方法检测RAGE含量。结果①与NC组比较,DM组大鼠体重、血糖、HbA1C、BUN、UAER、肾重/体重比值均明显增高(P<0.01),Ccr明显下降(P<0.01),肾脏AGEs及RAGE含量均明显增高(P<0.01);②与DM组相比,VE组UAER、肾重/体重比值明显降低(P<0.05),Ccr明显升高(P<0.01),肾脏AGEs及RAGE含量均明显降低(P<0.05)。结论维生素E无降糖作用,但可能通过减轻糖尿病大鼠肾组织的非酶糖基化过程,从而减轻AGEs与受体结合后导致的对肾脏组织的损伤,对糖尿病大鼠肾脏具有保护作用。     

苦参碱可抑制Th17所诱导的角质形成细胞IL-17RA、IL-21R及IL-22R1的表达

目的考察苦参碱(matrine,Mat)对Th17诱导的角质形成细胞(keratinocytes,KC)系HaCaT细胞白细胞介素-17受体A(IL-17RA)、白细胞介素-21受体(IL-21R)及白细胞介素-22受体1(IL-22R1)表达的影响。方法培养HaCaT细胞,用Th17分泌的主要效应因子IL-17A(10ng/mL)、IL-21(10ng/mL)及IL-22(10ng/mL)联合刺激HaCaT细胞模拟类银屑病样细胞模型,再用10μg/mL及100μg/mL的Mat干预此细胞模型,采用RT-qPCR及Western blot方法检测IL-17RA、IL-21R及IL-22R1的表达变化。结果 IL-17RA、IL-21R及IL-22R1的mRNA及蛋白在HaCaT细胞上有表达。IL-17A、IL-21及IL-22联合刺激可以显著促进HaCaT细胞IL-17RA、IL-21RmRNA及蛋白表达,IL-22R1蛋白的表达量升高(P<0.05)。Mat单独干预或与上述细胞因子联合干预组与未处理的HaCaT细胞组相比,IL-17RA、IL-21R及IL-22R1表达无显著下降(P>0.05),但可以显著降低IL-17A、IL-21及IL-22联合刺激后所升高的受体蛋白表达(P<0.05)。结论 IL-17A、IL-21及IL-22联合刺激角质形成细胞可以增强IL-17RA、IL-21R及IL-22R1的表达,Mat可抑制这种增强作用,可能在多种Th17介导的免疫性皮肤病中发挥抗炎作用。     

阻断5-HT1B受体对正常和帕金森病大鼠底丘脑核神经元活动的不同作用

目的观察体循环给予5-HT1B受体拮抗剂SB224289后,对正常大鼠和帕金森病(Parkinsons disease,PD)模型大鼠底丘脑核(subthalamic nucleus,STN)神经元电活动的影响。方法采用在体细胞外记录的方法,研究正常和PD大鼠STN神经元电活动的变化;比较阻断5-HT1B受体后两组大鼠STN神经元电活动的改变。结果①PD组大鼠STN神经元的平均放电频率[(7.87±1.37)Hz]明显高于正常组大鼠[(6.59±1.44)Hz,P<0.001]。两组大鼠STN神经元均表现为规则、不规则和暴发式放电3种形式。正常组大鼠具有规则、不规则和暴发式放电的神经元比例分别为33.33%、54.17%和12.50%;PD组分别为31.25%、35.42%和33.33%,PD组大鼠具有暴发式放电的神经元的百分比显著高于正常组(P<0.01)。②阻断5-HT1B受体后,正常大鼠STN神经元平均放电频率明显增高(P<0.01),具有暴发式放电的神经元数量显著增多(P<0.05)。而PD组大鼠阻断5-HT1B受体后STN神经元的放电频率和放电形式均无明显变化。结论 PD大鼠STN神经元呈过度兴奋状态;阻断5-HT1B受体可兴奋正常大鼠的STN神经元,而不影响PD大鼠的神经元活动。     

正常人及肠易激综合征患者肠黏膜孤啡肽受体的表达

目的检测正常人及肠易激综合征(IBS)患者肠黏膜孤啡肽受体(ORL-1)的表达,探讨其在IBS患者发病机制中的作用。方法黏膜标本取自20例正常人和27例腹泻型(D-IBS)、23例便秘型(C-IBS)患者空肠上段及升结肠,应用实时荧光定量RT-PCR法对肠黏膜ORL-1的表达进行定量分析。结果正常对照组空肠上段及升结肠均有ORL-1mRNA的表达,且空肠上段表达量高于升结肠(P<0.05);D-IBS患者空肠及升结肠黏膜ORL-1mRNA表达量均低于正常对照组(P<0.05),C-IBS患者与正常对照组间无差异。结论 ORL-1受体在人胃肠运动及IBS的病理生理机制中可能具有重要作用;OFQ/ORL-1系统对IBS不同亚型的调节具有异质性。     

溃疡性结肠炎大鼠血清酪酪肽水平及其受体特性的测定

目的 通过测定溃疡性结肠炎(UC)大鼠血清酪酪肽(PYY)水平及其空肠黏膜上皮细胞膜PYY受体的特性来观察UC时血清PYY水平及其受体的变化.方法 将大鼠随机分成UC组、腹泻型肠易激综合征(D-IBS)组及对照组,用同位素放射免疫法分别检测各组的PYY水平及反映受体特性的两个参数:平衡解离常数(Kd)和最大结合容量值(Bmax).结果 UC及D-IBS组血清PYY水平高于对照组(P<0.001),且UC组亦高于D-IBS组(P<0.001);但UC组Kd及Bmax值与正常组和D-IBS组比较均无明显差异(P>0.05).结论 UC组PYY水平高于正常及D-IBS组,推测UC时PYY水平的变化可能与其腹泻症状和炎症病变的产生有关;UC组的Kd及Bmax与正常组比较无显著变化,推测UC可能与PYY受体的变化无关.     

Induction of animal model of Graves' disease in BALB/c mice

Objective To construct an animal model of Graves' disease (GD) by immunizing BALB/c mice with hM12 cells co-expressing major histocompatibility complex (MHC) class Ⅱ molecules and human thyrotropin receptor (TSHR) molecules. Methods BALB/c mice in experimental group (H-2d) were immunized with hM12 cells Intraper-itoncally every 2 weeks for six times, while mice in control group were immunized with M12 cells. Five weeks later, the thyroids were histologically examined, and serum samples were tested for thyroid-stimulating antibodies (TSAb) and thyroid hormone levels. Results One BALB/c mouse in experimental group developed Graves'-like disease. Total T4 and T3 levels in this mouse were above the upper limit of normal, TSAb activity was displayed in its serum. The thyroid histologically showed the features of thyroid hyperactivity including thyrocyte hypercellularity and colloid absorption.None of control mice developed Graves'-like disease. Conclusion An animal model with some characteristics of human Graves' disease was successfully induced and the model will facilitate studies aimed directly at understanding the patho-genesis of autoimmunity in Graves' disease.     

Glutamate enhances the expression of vascular endothelial growth factor in cultured SD rat astrocytes

Objective To study the effect of glutamate on the expression of vascular endothelial growth factor (VEGF) mRNA and protein in cultured rat astrocytes. Methods Cultured rat astrocytes were randomly divided into 6 groups: control group (C), glutamate group (G), QA group (Q), DCG-IV group (D), L-AP4 group (L) and glutanmte-FMCPG gronp (G+M). Cells were cultured under nomoxic condition (95% air, 5% CO2). RT-PCR and ELISA methods were used to detect the expression of VEGF mRNA and protein in cultured astrocytes, respectively. G+ M group was preincubated with lmM MCPG for 30 min prior to the stimulation with glutamate. There were 7 time points at 0,4,8,12,16,24 and 48 h in each group except G+M group. Results The expression of VEGF mRNA and protein did not differ significantly among D group, L group and C group. Different from that in C group, the expression of VEGF mRNA and protein could be enhanced both in a dose-dependent and time-dependent manner in G group and Q group. Meanwhile, the enhanced expression of VEGF mRNA and protein in G group was completely suppressed by MCPG after 24 h. Conclusion Glutamate can increase the expression of VEGF mRNA and protein in cultured astrocytes, which may be due to the activation of group I metabotropic glutamate receptors in astrocytes.     

用流式细胞测定仪测定AITD患者TRAb

以人工重组ＴＳＨ受体表达的中国仓鼠卵细胞（ＣＨＯ－ＴＳＨＲ）为靶细胞,用流式细胞（ＦＡＣＳ）测定技术,对４５例自身免疫性甲状腺病（ＡＩＴＤ）患者血清中ＳＨ受体抗体（ＴＲＡｂ）进行测定。结果显示：３６例ＧＤ患者阳性细胞结合的百分数为１．９％～７７．４％。阳性率为９１．６６％。其均值２６．１３９％与正常人均值６．１６３％相比有显著性差异（Ｐ＝０．０００１）。９例桥本（ＨＴ）甲减患者５例阳性,阳性率为５５．５６％。证实以ＣＨＯ－ＴＳＨＲ细胞为靶细胞用ＦＡＣＳ进行ＴＲＡｂ汕测定不失为一种敏感简便的新方法。