重症肌无力伴胸腺瘤患者胸腺组织蛋白4.1R mRNA的表达研究

目的探讨蛋白4.1R在重症肌无力(myasthenia gravis,MG)伴胸腺瘤患者胸腺组织中的表达情况。方法运用半定量RT-PCR方法检测7例胸腺瘤MG与7例正常对照组胸腺组织中蛋白4.1R的表达。结果胸腺瘤MG组胸腺组织中蛋白4.1R mRNA的相对表达水平为0.84±0.46,与对照组0.43±3.69相比明显增高,差异有统计学意义(P0.05)。结论蛋白4.1R在胸腺瘤MG患者胸腺组织中mRNA水平表达异常,可能干扰胸腺细胞的信号传导,从而影响胸腺细胞的增殖和活化,参与了MG的发生。     

自身免疫调节因子在重症肌无力患者胸腺内的表达及作用分析

目的比较自身免疫调节因子(AIRE)在重症肌无力(MG)患者及正常人胸腺内的含量,分析AIRE在MG发病中的作用。方法收集濮阳市人民医院胸外科MG胸腺切除患者的胸腺组织及心脏外科手术患者的胸腺组织,将其分为MG合并胸腺异常组(A)、MG胸腺正常组(B)、对照组(C)。采用蛋白免疫印迹法及荧光定量PCR法比较3组胸腺内AIRE基因及蛋白含量。结果 C组(0.75±0.04)AIRE基因含量明显高于A(0.42±0.08)、B(0.45±0.08)两组,差异有统计学意义(P0.05);A、B两组之间AIRE基因含量差异无统计学意义(P0.05)。C组(0.72±0.05)AIRE蛋白含量较A(0.40±0.08)、B(0.50±0.05)两组高,差异有统计学意义(P0.05);A、B两组之间蛋白含量差异无统计学意义(P0.05)。结论 MG患者胸腺内AIRE基因及蛋白含量较对照组偏低,而与胸腺病理类型无关,AIRE在MG发病过程中有重要作用。     

GSTM3 mRNA在重症肌无力患者胸腺组织中的表达

目的 探讨谷胱甘肽S-转移酶m3(GSTM3mRNA在重症肌无力患者及正常人胸腺组织中的表达情况.方法 利用荧光定量聚合酶链反应(FQ-PCR)方法定量分析GSTM3 mRNA在重症肌无力胸腺与正常胸腺组织中的表达水平.结果 GSTM3 mRNA在重症肌无力患者和正常人胸腺组织中的表达水平分别为0.67±0.10和0.58±0.05,两者比较差异有统计学意义(P<0.01.结论 GSTM3 mRNA在MG胸腺组织中高表达,可能与MG的发生、发展密切相关.     

重症肌无力胸腺和外周血抗原特异性单个核细胞IFN-γ、IL-4的转录及表达

目的为了解重症肌无力(MG)胸腺细胞免疫状态,对10例行胸腺摘除(Tx)的MG患者及4例先天性心脏病行开胸手术患者的胸腺和外周血单个核细胞经特异性抗原乙酰胆碱受体(AChR)刺激后,其细胞IFN-γ和IL-4的mRNA转录和细胞培养上清液中的蛋白表达情况进行了检测.方法采用RT-PCR结合狭缝印迹杂交检测IFN-γ、IL-4 mRNA转录,采用ELISA检测IFN-γ和IL-4的表达情况.结果 10例MG患者的胸腺和外周血单个核细胞经AChR刺激后IFN-γ和IL-4的mRNA转录增高,其中8例胸腺增生者胸腺细胞和外周血细胞的IFN-γ mRNA转录、外周血细胞的IFN-γ表达均高于健康对照组(P＜0.05);胸腺细胞IL-4 mRNA转录和表达均高于健康对照,差异有显著性(分别为P＜0.01和P＜0.05),而外周血细胞IL-4 mRNA转录和表达与健康对照差异无显著性(P＞0.05);另2例伴胸腺瘤者,此两种细胞因子无论mRNA转录还是蛋白表达均很低.外周血细胞与胸腺细胞IL-4 mRNA转录有一定的相关性,基本反映胸腺的免疫学变化.结论胸腺增生MG患者的细胞免疫(Th1和Th2)高度活跃,PBMC的有些免疫学参数能反映MG患者胸腺的免疫学改变,但有待进一步累积更多资料.伴胸腺增生与伴胸腺瘤的MG发病机制可能不同,Tx是必要的.     

三磷酸腺苷结合盒运转体A1与颈动脉粥样硬化斑块的关系

目的 探讨三磷酸腺苷结合盒运转体A1(ATP binding cassette transporter A1,ABCA1)在人颈动脉粥样硬化斑块中的表达变化及作用机制.方法 收集24例人颈动脉粥样硬化斑块标本和10例肠系膜动脉标本(对照组),采用RT-PCR测定ABCA1 mRNA和视黄酸X受体α(RXRα)mRNA表达水平,并采用Western Blot检测ABCA1及RXRα的蛋白表达水平.24例人颈动脉粥样硬化斑块标本按病理分级,比较病理组织为Ⅲ级和Ⅰ级动脉粥样硬化组织间ABCA1 mRNA、RXRαmRNA表达水平及蛋白表达水平.结果 颈动脉粥样硬化斑块组的ABCA1 mRNA(0.79±0.04)和RXRα mRNA(0.73±0.04)表达与对照组相比上调,差异有统计学意义(P＜0.05);ABCA1 mRNA与RXRα mRNA增加水平相关(P＜0.05);颈动脉粥样硬化斑块的ABCA1蛋白表达(0.22±0.03)下调水平与对照组(0.53±0.03)相比差异有统计学意义(P＜0.05);Ⅲ级和Ⅰ级动脉硬化斑块ABCA1mRNA、RXRα mRNA及蛋白表达水平差异有统计学意义(P＜0.05).结论 ABCA1及RXRα蛋白表达水平下调可能是进展性动脉粥样硬化损害的关键因素.     

重症肌无力患者胸腺细胞Bcl-2表达的研究

目的 探讨重症肌无力(myasthenia gravis,MG)的发病机制,为临床治疗提供一定的实验理论依据.方法 用机械分离法、淋巴细胞分离液获取19例MG患者和11例正常对照组胸腺细胞悬液,接种于培养板中,分别加入雌、孕激素作为特异性刺激组,未加内分泌激素作为空白对照组,培养72h后收获细胞.采用流式细胞术分别检测MG患者和正常对照组胸腺细胞及雌、孕激素培养前后胸腺细胞的Bcl-2表达.结果 MG患者胸腺细胞Bcl-2袁达率比正常对照组显著增高,2者相比有统计学意义(P＜0.01);雌、孕激素培养组MG患者胸腺细胞Bcl-2的表达率比空白对照组显著降低,分别比较差异有统计学意义(P＜0.05),雌激素培养组与孕激素培养组比较差异无统计学意义(P＞0.05).结论 MG患者胸腺细胞Bcl-2表达升高,可能参与调控胸腺细胞凋亡,其表达异常可能是MG发病的一个重要因素;雌、孕激素抑制MG患者胸腺细胞Bcl-2的表达,提示体内雌激素、孕激素水平变化与MG患者病情变化有一定联系.     

丹参素预处理对脑梗死大鼠血浆纤溶激活系统的影响

目的 观察丹参素预处理对脑梗死大鼠血浆纤溶激活系统的影响.方法 40只雄性大鼠,随机分为假手术组、脑梗死对照组、低剂量[5 mg/(kg·d)]丹参素组和高剂量[10 mg/(kg·d)]丹参素组,每组10只.应用自体血栓注入,制成大鼠脑梗死模型;用ELISA方法 检测血浆中组织型纤溶酶原激活物(t-PA)及其抑制物(PAI-I)含量;RT-PCR测定大鼠脑组织中t-PA mRNA及其PAI-I mRNA表达的变化.结果 与脑梗死对照组[(2.65±0.43)ng/ml]比较,低剂量及高剂最丹参素组t-PA含量[(3.29±0.13)ng/ml、(6.37±0.19)ng/ml]明显升高(均P＜0.01),两丹参素组之间差异亦有统计学意义(P＜0.0I);与脑梗死对照组比较,低剂量丹参素组PAI-I含量[(2.65±0.40)ng/ml]降低(P＜0.05),高剂量丹参素组PAI-I含量[(1.98±0.32)ng/ml]明显降低(P＜0.01),两丹参素组之间差异亦有统计学意义(P＜0.01);与脑梗死对照组比较,低剂量及高剂量丹参素组t-PA mRNA表达[灰度值分别为(0.37±0.14)、(0.46±0.12)]均明显升高(均P＜0.01),两丹参素组之间差异亦有统计学意义(P＜0.01);与脑梗死对照组比较,低剂量及高剂量丹参素组PAI-I mRNA表达[灰度值分别为(0.54±0.12)、(0.36±0.07)]均明显降低(均P＜0.01),两丹参素组之间差异亦有统计学意义(P＜0.01).结论 丹参素可以改善脑梗死大鼠纤溶功能,并且有剂量依赖性.     

胸腺瘤患者胸腺淋巴细胞Fas表达与重症肌无力的相关性

目的 探讨胸腺瘤患者发生重症肌无力(myasthenia gravis,MG)的机制及Fas基因在MG发病中的作用.方法 收集131例胸腺瘤患者,按是否合并MG分为合并MG组(70例)和不合并MG组(61例)两组.应用免疫组织化学、蛋白电泳、聚合酶链反应和酶联免疫吸附等方法检测两组患者胸腺淋巴细胞Fas蛋白表达和血清可溶性Fas(sFas)水平,应用DNA测序技术检测Fas基因结构变异.结果 胸腺瘤合并MG组患者胸腺淋巴细胞Fas表达水平明显低于胸腺瘤不合并MG组,sFas含量[(3879.06±706.51) pg/mL]明显高于胸腺瘤不合并MG组[(1868.18±391.46) pg/mL] (P＜0.01);胸腺瘤合并MG组胸腺淋巴细胞Fas基因第6外显子16和21位点T→G置换突变发生率分别为65％和75％,明显高于胸腺瘤不合并MG组(两位点均为5％)(P＜0.05).结论 胸腺淋巴细胞Fas基因外显子6某些位点发生突变,所编码的Fas蛋白跨膜区变异或缺乏跨膜区而导致Fas结构和功能的改变可能是MG发生的主要原因.     

重症肌无力胸腺切除术免疫功能变化及疗效观察

目的 观察重症肌无力胸腺切除术免疫功能变化及临床疗效.方法 选取84例重症肌无力患者为研究对象,胸腺切除术治疗患者52例为手术组,胸腺未切除患者32例为非手术组,同时选取健康志愿者30例为对照组.观察胸腺切除手术免疫功能的变化及临床疗效.结果 非手术组患者外周血AchRab水平(2.02±0.28) ng/L显著高于对照组(1.58±0.32)ng/L,差异有统计学意义(P＜0.05);手术组患者外周血AchRab水平(1.72±0.28)ng/L高于对照组,同时低于非手术组,差异有统计学意义(P＜0.05);3组免疫球蛋白水平差异无统计学意义(P＞0.05).胸腺切除术患者临床总有效率81.8％,明显高于非手术组的60.7％,差异有统计学意义(P＜0.05).结论 通过手术免疫功能变化观察,重症肌无力与胸腺密切相关,术前MG患者免疫功能处于紊乱状态,手术切除胸腺后免疫功能可得到一定程度恢复和改善.     

孕期酒精暴露对子代大鼠学习记忆及海马N-甲基-D-天冬氨酸受体2B亚基表达的影响

目的 研究孕期酒精暴露对子代大鼠学习记忆及海马N-甲基-D-天冬氨酸(NMDA)受体2B亚基(NR2B)表达的影响.方法 按照随机数字表法,将雌性SD大鼠随机分为正常对照组、饮酒对照组和孕期酒精暴露组,每组各8只;饮酒法建立大鼠孕期酒精暴露模型,子代成年后,采用Y-型迷宫测试子鼠学习记忆成绩;采用聚合酶链反应分析子鼠海马组织NR2B mRNA的表达;采用免疫荧光法检测子鼠海马区NR2B蛋白表达.结果 (1)各组子鼠成年后学习记忆成绩的差异有统计学意义(F=4.566,P＜0.05),孕期酒精暴露组子鼠学习记忆成绩[(43.00±15.33)次]比正常对照组[ (25.13±12.35)次]和饮酒对照组[(26.12±11.95)次]明显下降(P均＜0.05);(2)各组子鼠成年后海马组织中NR2B mRNA表达差异有统计学意义(F=29.795,P＜0.01),孕期酒精暴露组子鼠海马区NR2B mRNA表达(0.97±0.14)较正常对照组(0.52±0.10)和饮酒对照组(0.62±0.12)明显上升(P均＜0.01);孕期酒精暴露组子鼠海马区NR2B蛋白表达明显增加.结论 孕期酒精暴露对子代大鼠的神经损伤可能与NMDA受体亚基NR2B蛋白表达的上调有关.     

Mechanisms of the suppression of the nociceptive reflex of the opening of the mouth during stimulation of the structures of the limbic brain

The comparative effectiveness of the inhibitory influence of tetanic stimulation of hypothalamus, amygdala and limbic cortex on EMG-response of m. digastricus evoked by electrical stimulation of tooth pulp nociceptive afferents was studied in cats anesthetized with a mixture of chloralose and nembutal. It was found that inhibition of the EMG-component of the jaw-opening reflex is most pronounced in case of stimulation of medial and lateral region of the hypothalamus, the inhibitory effect of central and medial nuclei of the amygdala is less pronounced and the effect of the limbic cortex is the weakest. It was shown that the mechanism of the antinociceptive effect of tetanic stimulation of the hypothalamus is not related to the concomitant increase of the blood pressure. After stabilization of the blood pressure the suppressive effect of the hypothalamus remains without changes, that points out to a direct, primary, not baro-afferent mechanism of the inhibition of the activity of nociceptive neurons of the trigeminal sensory nuclei. Noradrenaline, injected intravenously, induced a large increase of the blood pressure accompanied by a pronounced inhibition of the pain reflex. Angiotensin causes the same degree of blood pressure elevation without changes in the amplitude of the EMG-response of the pain reflex. Hypothalamic and noradrenergic mechanisms for control of pain sensitivity are discussed.     

药物治疗与合并认知行为治疗对强迫症疗效的比较

目的探讨认知行为心理治疗(CBT)在强迫症(OCD)患者各亚型治疗中的有效性和规律性。方法本研究为临床对照研究。符合入组标准的强迫症患者按患者自愿原则分为两组,治疗观察3、6、12个月。疗效评定分别运用Yale-Brown强迫量表,自拟的自评好转程度量表和临床疗效评定。结果认知行为心理治疗合并药物治疗组31例,临床有效率70.9%,其中治愈率1.8%。单纯药物治疗组24例,临床有效率33.3%。Yale-Brown强迫量表和自评量表得分在6个月和12个月两组有显著差异(P<0.05)。其中强迫症亚型(怕脏型、反复检查型和反复担心型)的疗效比较,怕脏型在治疗3个月末两组间自评量表评分有显著性差异(P<0.05);反复担心型在治疗6个月末两组间Yale-Brown强迫量表总分有显著性差异(P<0.05);反复检查型两组间无统计学差异。结论认知行为心理治疗合并药物治疗强迫症的疗效明显优于单纯药物治疗。强迫症的亚型在治疗中的有效性次序为:反复担心型>怕脏型>反复检查型。     

Characteristics of the sympathetic innervation of the nictitating membrane and of the vasculature of the nose and tongue of the cat

Summary Vasomotor responses from the nasal mucosa and tongue, and contractions of the nictitating membrane, were recorded on stimulation of the cervical sympathetic or internal carotid nerves.Preganglionic sympathetic nerve fibres which elicited a membrane response possessed a lower threshold than those which evoked nasal vasoconstriction, while the latter displayed a lower threshold than fibres which evoked tongue vasoconstriction. The sympathetic vasodilator fibres to the tongue, whose activity was revealed after-receptor blockade, had a similar threshold to the vasoconstrictor fibres.Membrane contraction, nasal vasoconstriction and occasionally tongue vasoconstriction could be evoked by stimulating the internal carotid nerve. The postganglionic fibres innervating the nasal mucosa had a similar threshold to those of the nictitating membrane, which may indicate that there are small myelinated fibres innervating the mucosa.The preganglionic compound nerve action potential had four major components, S₁–S₄. S₁, S₂ and usually S₃ fibres were associated with membrane contraction; S₂, S₃ and sometimes S₁ fibres were associated with nasal vasoconstriction; and S₃, usually S₂ and occasionally S₁ fibres were associated with vasoconstriction in the tongue. It is concluded that each of these three groups of nerve fibres, but not S₄ fibres, may include fibres associated functionally with the three effectors.There was a considerable difference between the relative amplitude of the responses of the three effectors elicited by stimulation of the cervical sympathetic nerve at frequencies between 0.2 and 2 Hz. Vasoconstrictor responses were relatively larger than membrane contractions suggesting differences in the mechanisms of neurotransmission at the neuroeffector junctions.     

Development of the projection from the nucleus of the brachium of the inferior colliculus to the superior colliculus in the ferret

Neurons in the deeper layers of the superior colliculus (SC) have spatially tuned receptive fields that are arranged to form a map of auditory space. The spatial tuning of these neurons emerges gradually in an experience-dependent manner after the onset of hearing, but the relative contributions of peripheral and central factors in this process of maturation are unknown. We have studied the postnatal development of the projection to the ferret SC from the nucleus of the brachium of the inferior colliculus (nBIC), its main source of auditory input, to determine whether the emergence of auditory map topography can be attributed to anatomical rewiring of this projection. The pattern of retrograde labeling produced by injections of fluorescent microspheres in the SC on postnatal day (P) 0 and just after the age of hearing onset (P29), showed that the nBIC-SC projection is topographically organized in the rostrocaudal axis, along which sound azimuth is represented, from birth. Injections of biotinylated dextran amine-fluorescein into the nBIC at different ages (P30, 60, and 90) labeled axons with numerous terminals and en passant boutons throughout the deeper layers of the SC. This labeling covered the entire mediolateral extent of the SC, but, in keeping with the pattern of retrograde labeling following microsphere injections in the SC, was more restricted rostrocaudally. No systematic changes were observed with age. The stability of the nBIC-SC projection over this period suggests that developmental changes in auditory spatial tuning involve other processes, rather than a gross refinement of the projection from the nBIC.     

A contribution to the study of the architecture of the autonomic nervous system of the digestive tract of the rat

Summary The distribution of aminergic and non-aminergic nerve fibres to the different constituents of the wall of the digestive tract in various regions is described. Aminergic fibres synapse with all nervous perikarya. Densely interlacing networks of nerve fibres are found in both layers of the tunica muscularis and in the lamina muscularis mucosae. A finely meshed plexus is observed in relation to the wall of the blood vessels in the wall of the gut. There are many fibres connecting the muscular and the vascular plexus. No nerve fibres have been observed in direct relation to the epithelium.The functional implications of these findings are discussed.