自身免疫性心肌炎小鼠心肌组织病理学及酶学指标分析

目的建立小鼠自身免疫性心肌炎模型,观察心肌组织学、酶学指标的变化。方法选择遗传易感的雄性Balb/c小鼠,于初次免疫后的第7、30d多点皮下重复注射猪心肌肌球蛋白,分别于免疫后的第21、63d处死小鼠,观察心肌组织病理变化情况;测定血清乳酸脱氢酶（LDH）、肌酸激酶（CK）、肌酸激酶同工酶（CK-MB）等心肌酶学指标。结果第21d,实验组小鼠呈现急性心肌炎表现,心外膜下、心肌间质单个核细胞浸润明显,心外膜下较心肌间质病变更为显著,心肌酶谱各项指标明显高于正常对照组（P〈0.05）。第63d,小鼠呈慢性心肌炎表现,炎症细胞浸润减少,心肌纤维化明显,单个核细胞浸润心肌间质较心外膜显著,心肌酶谱各项指标较急性期有所下降,但仍高于正常对照组（P〈0.05）。结论应用猪心肌肌球蛋白可成功建立小鼠自身免疫性心肌炎模型。     

自身抗体在自身免疫性肝病中的表达及临床价值

目的研究自身抗体在自身免疫性肝病患者中的表达及临床价值。方法间接免疫荧光法检测自身免疫性肝病患者血清中的自身抗体。结果 36例AIH患者抗平滑肌抗体（ASMA）阳性率58.33%,抗肝肾微粒体抗体Ⅰ型（LKM-1）阳性率5.56%,抗可溶性肝抗原抗体（SLA）阳性率2.78%和抗线粒体抗体Ⅱ型（AMA-M2）阳性率8.33%;48例PBC患者抗线粒体抗体Ⅱ型（AMA-M2）阳性率97.92%,其他抗肝抗原自身抗体均阴性。AIH和PBC患者抗核抗体（ANA）阳性率分别为77.77%、100%。5例PSC患者ANA阳性率60%,抗肝抗原自身抗体均为阴性。31例健康体检者ANA只检测1例阳性,抗肝抗原自身抗体均为阴性。自身免疫性肝病各组阳性检出率与健康对照组之间比较差异具有统计学意义（P〈0.01）。结论抗核抗体（ANA）及抗肝抗原自身抗体的检测对自身免疫性肝病的诊断和分型具有重要价值。     

童心康合剂对实验性自身免疫性心肌炎大鼠心肌纤维化的影响

目的:探讨童心康合剂对实验性自身免疫性心肌炎(EAM)模型大鼠心肌纤维化的影响。方法:将60只Lewis大鼠随机分为正常对照组、EAM模型组、玉丹荣心丸组、童心康合剂高、中、低剂量组,每组10只。于免疫56d处死存活大鼠,观察心肌病理变化、Masson染色观察心肌纤维化,免疫组化测心肌组织金属蛋白酶-9(MMP-9)及其组织抑制物-1(TIMP-1)的表达。结果:童心康合剂大中剂量组与玉丹荣心丸组心肌病理积分均低于EAM模型组(P<0.05),3组之间差异无显著性(P>0.05),童心康合剂小剂量组与EAM模型组病理积分差异无显著性(P>0.05)。童心康合剂大中剂量、玉丹荣心丸均能抑制心肌纤维化,童心康合剂大中剂量均优于玉丹荣心丸(P<0.05),且2组之间差异无显著性(P>0.05)。童心康合剂中剂量能上调心肌组织MMP-9的表达,下调其TIMP-1的表达。结论:童心康合剂大中剂量、玉丹荣心丸可减轻EAM模型大鼠心肌炎症,抑制心肌纤维化,此作用可能与调节MMP-9/TIMP-1平衡有关。     

自身免疫性心肌炎大鼠心肌病理改变与心肌标志物水平的相关性

目的 探讨N端前体脑钠肽(NT-proBNP)、肌钙蛋白I(cTnI)、肌酸激酶同工酶MB(CK-MB)在心肌炎诊断中的价值.方法 24只雄性Lewis大鼠完全随机分为对照组(n=12)和模型组(n=12).对照组大鼠皮下注射完全弗氏佐剂,模型组皮下注射心肌球蛋白诱导形成自身免疫性心肌炎.在初次免疫后第21天,进行心脏超声检测、血清cTnI、CK-MB、NT-proBNP水平检测及心肌组织病理学检查,并评价心肌炎变的严重程度,进行宏、微观病理评分.通过Logistic回归分析心肌炎病理评分与心肌标志物水平的相关性.结果 与对照组比较,心肌炎模型组大鼠左心室射血分数明显减弱[(65.8±4.4)％比(86.9±1.1)％,P＜0.05],收缩末期内径明显增大[(3.88±0.29) mm比(3.16±0.08)mm,P＜0.05];CK-MB、cTnI及NT-proBNP都明显升高[( 1357±397)IU/L比(392±120)IU/L,(17.2±5.3)μg/L比(6.8±1.8) μg/L,(706± 218) ng/L比( 110±46) ng/L,均P＜0.05].CK-MB与心肌病理宏观评分、微观评分均无明显相关性(P＞0.05),cTnI与心肌病理微观评分明显相关(P＜0.05),NT-proBNP与心肌宏观形态病理改变明显相关(P＜0.05).结论 CK-MB在一定程度上提示心肌损伤,但不能作为评价心肌炎症严重程度的指标;cTnI能够较好地提示心肌炎症细胞水平的严重程度,为心肌炎组织学水平病理改变的良好指标;NT-proBNP能够在形态学水平评价整体心肌损害的严重程度,但是在组织学水平其评价能力欠佳.     

自身免疫性肝病患者血清中自身抗体表达及诊断价值探讨

目的 探讨自身抗体在自身免疫性肝病中的表达及其在临床诊断作用.方法 采用免疫印迹法:检测AMA-M2、M2-3E、SP100、PML、GP-210.ELISA法检测ANA及各类病毒性肝炎标志物.结果 血清自身抗体ANA及AMA-M2、M2-3E、SP100、PML、GP21O的结果在自身免疫性肝炎分别为PBC组37.7%、79.2%、81.1%、18.9%、22.6%、20.8%,AIH组为61.2%、8.2%、4.1%、4.1%、4.1%、2.0%,HBV组为6.7%、0、0、0、0、0,正常对照组为2.0%、0、0、0、0、0.结果分析显示,PBC组各指标均显著高于HBV组和对照组(P＜0.01),AIH组ANA也显著高于HBV组和对照组(P＜0.01).结论 自身抗体的检测对自身免疫性肝病具有较高诊断价值.     

实验性自身免疫性葡萄膜视网膜炎小鼠模型的建立及观察

目的：研究实验性自身免疫性葡萄膜视网膜炎（EAU）的小鼠模型的发生规律及特征。方法6-8周B10.RⅢ小鼠,用人源性IRBP肽161-180与完全弗氏佐剂混合注射于小鼠建立模型。观察小鼠眼部表现,小鼠眼球病理切片,光镜观察记录不同时间点小鼠的炎症情况。结果 EAU小鼠视网膜炎症出现在第14-21天,表现为结膜充血,前房混浊,虹膜后粘连,眼底视网膜水肿充血。病理切片见玻璃体腔炎症细胞,视网膜水肿,各层结构紊乱,炎症浸润,血管周围炎,网膜内肉芽肿等。结论用IRBP161-180肽成功诱导B10.RⅢ鼠建立EAU模型,为人类葡萄膜炎的研究奠定了良好的实验基础。     

免疫学指标对自身免疫性肝病的诊断意义

自身免疫性肝脏疾病主要包括原发性胆汁性肝硬化(primary biliary cirrhosis,PBC)、自身免疫性肝炎(autoimmune hepatitis,AIH)、原发性硬化性胆管炎(primary sclerotic cholangitis,PSC)),以及这三种疾病中任何两者之间的重叠综合征。虽然它们均属自身免疫性肝病,但病因、临床表现、及治疗各不相同,而早期诊断和早期治疗又是控制疾病进展的关键,故诊断及鉴别诊断成为临床急需解决的问题。其中实验室检察特别是免疫指标的测定有着重要的作用。1资料与方法1.1研究对象2000年1月至2006年12月在我院门诊及住院的肝病患者200例,其中:①原发…     

治疗自身免疫性心肌炎药物实验研究进展

自身免疫性心肌炎进展快，复发率高，可导致严重心律失常、心源性休克和充血性心力衰竭等。临床上还没有确切有效的药物，主要以对症处理为主。近年来国内外从不同的角度开展了大量关于治疗该病药物的实验研究，如抑制炎症因子或共刺激分子的基因靶向治疗、调节氧化还原反应药物、调节Th1／Th2炎症因子分泌药物、抑制心肌重构药物的探索与开发等。现就治疗该病药物实验研究进展予以综述，以指导药物临床试验与应用，并为未来该领域讲一步研究提供参考。     

肝病相关自身抗体检测在自身免疫性肝病诊断中的价值研究

目的 自身免疫性肝病临床流行病学调查,观察各肝病患者中自身抗体检测的阳性率,自身免疫性肝病检测的阳性率;探讨自身抗体检测对自身免疫性肝病的诊断价值.方法 对2007年10月至2010年10月2 714份自身抗体检验结果进行回顾性分析,并对自身抗体阳性病例的查阅临床资料.结果 2 714份血清中自身抗体阳性472例,阳性率17.4％,抗核抗体(ANA)阳性418例,阳性率15.4％,抗线粒体抗体(AMA)阳性124例,阳性率4.57％,抗平滑肌抗体阳性91例,阳性率3.35％,抗肝肾微粒体抗体( LKM)阳性16例,阳性率0.59％,抗肝细胞膜抗体阳性(LMA)8例,阳性率0.29％,抗肝细胞溶质抗原1型抗体(LC-1)阳性5例,阳性率0.18％,抗肝特异性脂蛋白抗体(LSP)阳性3例,阳性率0.11％,2 914例送检标本中诊断为AIH 59例,阳性率为2.17％,诊断为PBC 61例,阳性率为2.24％.自身抗体阳性患者74.57％诊断为病毒性肝炎及相关疾病.结论 自身抗体检测是诊断自身免疫性肝病的必要条件,但这些自身抗体也可见病毒性肝炎,药物性肝炎及其他疾病.     

实验性小鼠病毒性心肌炎组织中连接蛋白43的表达及与心律失常的关系

目的探讨病毒性心肌炎时连接蛋白43含量和分布的变化及其和心律失常的关系.方法Balb/c小鼠35只随机分为2组,应用免疫组织化学SABC法对正常小鼠和实验性病毒性心肌炎小鼠心肌细胞连接蛋白43表达进行检测,并对小鼠心电图进行了观察和改良Curtis and Ravingerovand评分.结果连接蛋白43在正常小鼠心肌闰盘中分布均匀,病毒性心肌炎时连接蛋白43的分布面积和密度明显减弱甚至阴性;病毒性心肌炎小鼠较正常小鼠出现较明显的心律失常.结论病毒性心肌炎时受累的心肌细胞连接蛋白43的表达受到抑制可能是导致心肌细胞缝隙连接通讯障碍以致心律失常的一个原因.     

Amiodarone minimizes experimental autoimmune myocarditis in rats

Amiodarone, a promising drug for the treatment of tachyarrythmias, was recently found to have immunomodulatory effects in vitro. We hypothesized that amiodarone would affect the immune system in vivo and examined the effect of amiodarone on myocarditis in rats. We induced experimental autoimmune myocarditis in rats by cardiac myosin immunization and treated the animals with an intraperitoneal injection of amiodarone at 25 mg/kg/every other day, 10 times after the induction of experimental autoimmune myocarditis. In the treated group, both microscopic and macroscopic examinations showed reduced heart weights, a mild and localized infiltration of inflammatory cells and fibrosis in the myocardium, and a mild congestion in the liver and lungs as compared with the control group. The phenotypic distribution of lymphocytes in peripheral blood showed a significant decrease in the CD4/CD8a ratio in the treated group, but not in the control group. The proportion of mast cells involved in inflammatory cell infiltration was lower in the treated group than the control group. In vitro, amiodarone inhibited the proliferation of mast cells by arresting them in the G2 phase of the cell cycle. These results indicated that amiodarone minimized the progression of experimental autoimmune myocarditis, suggesting a potential therapeutic role for amiodarone treatment in patients suffering from myocarditis, especially myocarditis complicated by cardiac arrhythmias. One possible mechanism by which amiodarone minimizes the progression of experimental autoimmune myocarditis may be to affect the immune system via the immunomodulatory effects on T cell and mast cell functions.     

IL-13和IgG在慢性肺源性心脏病的表达及临床意义

目的探讨白细胞介素-13(IL-13)和免疫球蛋白G(IgG)在慢性肺源性心脏病(肺心病)发病及治疗中的意义。方法用ELISA法分别测定30例肺心病急性加重期患者、16例肺心病缓解期患者及15例正常对照者血清IL-13水平;用比浊法测定其IgG水平。结果(1)肺心病急性加重期组和缓解期组IL-13均高于正常对照组(P<0.01,P<0.05),肺心病急性加重期组IL-13水平高于缓解期组(P<0.05);(2)肺心病急性加重期组IgG水平显著高于缓解期组(P<0.05)和正常对照组(P<0.05),缓解期组与正常对照组比较差异无统计学意义(P>0.05);(3)肺心病急性加重期组IL-13和IgG水平呈直线正相关(r=0.90,P<0.001)。结论IL-13与肺心病的发病有关,且和IgG一起与导致肺心病急性加重的感染有关。     

Regulation of inflammation and myocardial fibrosis in experimental autoimmune myocarditis

Autoimmune responses and inflammation are involved in the pathogenesis of many cardiovascular diseases. There is compelling evidence that inflammatory mechanisms may contribute to progressive heart failure. Thus, myocardial infiltration of lymphocytes and mononuclear cells, increased expression of pro-inflammatory chemokines and cytokines and circulating autoantibodies are frequently observed in myocarditis and dilated cardiomyopathy (DCM). Experimental autoimmune myocarditis (EAM) in rodents may be elicited by immunization of cardiac myosin and EAM in rats mimics human fulminant myocarditis in the acute phase and human DCM in the chronic phase. Our animal model, EAM was demonstrated to progress into the clinicopathological state similar to DCM in the chronic phase, and was found to be characterized by the enlargement of the heart, dilatation of ventricles, diffuse and extensive myocardial fibrosis, besides being a cellular immunity and inflammation mediated disease. Severity of myocarditis was characterized by increased inflammation, cardiac fibrosis and decreased myocardial performance in rats with DCM. Pharmacological interventions such as angiotensin converting enzyme inhibitors (ACEI) and angiotensin receptor blockers (ARBs) significantly attenuated the myosin-induced inflammation and cardiac fibrosis and thereby improving myocardial function in rats with DCM. A growing body of evidence shows that ACEI and ARBs contribute to the pharmaceutical management of patients with heart failure mediated by immune and inflammatory response. The purpose of this review is to emphasize the role of inflammation and myocardial fibrosis in rats with DCM after EAM and study the effects of pharmacological interventions such as ACEI, ARBs in the treatment of heart failure through the suppression of inflammatory cytokines and fibrosis.     

直肠癌组织中CD70蛋白的表达及意义

目的探讨直肠癌组织中CD70(TNFSF7)蛋白的表达及其临床意义。方法采用免疫组化SP法检测60例直肠癌组织及癌旁组织中CD70、CD27(TNFRSF7)蛋白的表达,分析其与临床病理学特征的关系。结果 CD27、CD70在直肠癌组织中的阳性表达率分别为1.67%、33.33%;在直肠癌旁组织中均不表达。结论 CD70与直肠癌的发生发展密切相关,可作为判断肿瘤分期、指导治疗和预后判断的参考指标。     

Oleuropein prevents the development of experimental autoimmune myocarditis in rats

Oleuropein (OLE) is a natural secoiridoid that is derived from Olea europaea. OLE possesses cardioprotective effects in experimental models of hypertension, myocardial infarction, atherosclerosis and hyperlipidaemia. In the present study, the effects of OLE on experimental autoimmune myocarditis (EAM) were evaluated. EAM in rats were induced by subcutaneous injections of porcine cardiac myosin. Cardiac function parameters, myocardial pathology, myocardial inflammatory cell infiltration and nuclear factor kappa-B (NF-κB) expression were measured. Our data showed that the postmyocarditis rats exhibited increased left ventricular end systolic diameters, left ventricular end diastolic diameters, left ventricular end-diastolic pressures (LVEDP), and decreased ejection fractions. However, OLE significantly suppressed these changes in EAM rats. Histological analysis revealed that myosin induced miliary foci of discolouration on endocardial surfaces and extensive myocardial injuries with inflammatory cell infiltration were significantly improved by OLE therapy. A definitive positive correlation between the histological scores and LVEDP was observed. Moreover, OLE inhibited CD4⁺, CD8⁺ cells and macrophage infiltration in myocardium and decreased the serum production of tumour necrosis factor-a (TNF-a), interleukin-1β (IL-1β) and IL-6 in EAM rats. Expectedly, the myocardial levels of NF-κB p65, p-IκBa, IKKa were significantly attenuated by OLE, indicating the inhibitory effects of OLE on the NF-κB pathway. Furthermore, OLE decreased the myocardial expressions of phosphorylated-p38 MAPK, phosphorylated-ERK, and did not change the levels of p38 MAPK and ERK in EAM rats. Collectively, our results suggest that OLE effectively prevents the development of myocarditis, at least in part, by inhibiting the MAPKs and NF-κB mediated inflammatory responses.     

Cytotoxicities and Quantitative Structure Activity Relationships of B13 Sulfonamides in HT-29 and A549 Cells

B13 analogues are being considered as therapeutic agents for cancer cells, since B13 is a ceramide analogue and inhibits ceramidase to promote apoptosis in cancer cells. B13 sulfonamides are assumed to have biological activity similar to B13, since they are made by bioisosterically substituting the carboxyl moiety of B13 with sulfone group. Twenty B13 sulfonamides were evaluated for their in vitro cytotoxicities against human colon cancer HT-29 and lung cancer A549 cell lines using MTT assays. Replacement of the amide group with a sulfonamide group increased cytotoxicity in both cancer cell lines. The sulfonamides with long alkyl chains exhibited activities two to three times more potent than that of B13 and compound (15) had the most potent activity with IC(50) values of 27 and 28.7μM for HT-29 and A549, respectively. The comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) were used to carry out QSAR molecular modeling of these compounds. The predictive CoMSIA models for HT-29 and A549 gave cross-validated q2 values of 0.703 and 0.830, respectively. From graphical analysis of these models, we suppose that the stereochemistry of 1,3-propandiol is not important for activity and that introduction of a sulfonamide group and long alkyl chains into B13 can increase cytotoxicity.     

组织蛋白酶B在乳腺癌中的表达及其意义

目的探讨乳腺癌组织中组织蛋白酶B(CB)蛋白的表达及其意义。方法采用免疫组化S-P法检测50例纤维囊性乳腺病组织、65例乳腺浸润性导管癌组织中CB蛋白表达情况,并分析其与淋巴结转移的关系。结果 CB蛋白在浸润性导管癌中的阳性表达率为87.7%(57/65),明显高于在纤维囊性乳腺病中的表达,差异有统计学意义(P<0.05);且乳腺癌患者中有淋巴结转移组CB的表达明显高于无淋巴结转移组(P<0.05)。结论 CB蛋白的异常表达与乳腺浸润性导管癌的发生、发展及浸润转移有关。     

B7-H4在胃腺癌中的表达及临床意义

目的研究B7-H4蛋白在胃腺癌中的表达及临床意义。方法应用S-P免疫组化法检测B7-H4蛋白在20例正常胃黏膜、50例胃腺癌中的表达情况。结果 B7-H4在胃癌中的阳性表达率（82%）高于正常胃黏膜组（10%）,差异有显著意义（P〈0.05）。B7-H4在胃腺癌中的表达与年龄、性别无关（P〉0.05）,而与组织学分级、浸润深度及淋巴结转移有关（P〈0.05）。结论 B7-H4在胃腺癌中表达上调,可能与胃腺癌的转移和预后相关。     

Fusidin ameliorates experimental autoimmune myocarditis in rats by inhibiting TNF-alpha production

Experimental autoimmune myocarditis (EAM) represents a model for human autoimmune myocarditis, a condition for which no optimal treatment is currently available. It has been reported that tumor necrosis factor-alpha (TNF-alpha) plays a crucial role in pathogenesis of EAM. The immunomodulating antibiotic fusidic acid and its sodium salt (sodium fusidate-fusidin) were previously shown to reduce TNF-alpha production and its end-organ cytotoxicity, thus proving beneficial in several animal models of organ-specific autoimmune diseases. To investigate the effects of fusidin on EAM the drug was given at dose 80 mg/kg i.m. to EAM rats. Fusidin was administered as an early, from day 0 to 10, or late treatment, from day 10 to 21, after induction of disease. Both early and late treatment with fusidin markedly ameliorated the clinical and histological signs of the disease. Fusidin-treated rats had significantly decreased blood levels of TNF-a compared with vehicle-treated animals. Similarly, TNF-alpha production by in vitro sensitized lymph node cells in both fusidin treated groups was significantly lower than that in EAM rats. The present findings suggest that fusidin ameliorated EAM, at least partly, through an inhibitory action on the secretion of TNF-alpha.