淋巴细胞转输对线粒体腺苷酸转位酶诱导的自身免疫性心肌病小鼠细胞因子的抑制作用

目的:研究淋巴细胞转输对线粒体腺苷酸转位酶(ANT)合成肽诱导的自身免疫性心肌病传染性耐受的小鼠Th1/Th2亚群及血清和心肌组织细胞因子的影响。方法:用ANT多肽多次免疫Balb/C小鼠得到心肌病组,单抗组小鼠在给予ADP/ATP肽免疫的第0、1和2天,同时接受尾静脉注射400μg抗L3T4单抗;第6个月末时,将单抗组小鼠的脾细胞取出转输到正常小鼠体内(转输组),此小鼠亦同时给予多肽免疫,方法同心肌病组。对照组以不含ANT的相同免疫进行假性免疫,时间和剂量同心肌病组。采用流式细胞术检测脾T细胞内细胞因子IFN-γ/IL-4的含量;以ELISA法检测其血清中IFN-γ、白细胞介素-2(IL-2)、IL-4、IL-6和肿瘤坏死因子(TNF-α)水平;实时荧光定量PCR法检测其心肌细胞因子mRNA表达。结果:转输组Th细胞IFN-γ、IL-4含量与对照组和单抗组接近且明显低于心肌病组;转输组小鼠血清IFN-γ和IL-2水平明显高于心肌病组而低于单抗组,IL-4和IL-6水平显著低于心肌病组;TNF-α水平在转输组则最高;心肌细胞因子mRNA的表达则转输组显著低于心肌病组,与对照组和单抗组相近。结论:淋巴细胞转输能够阻断心肌病诱导过程中绝大部分细胞因子的生成,与单抗早期干预的结果类似。     

扩张型心肌病的早期干预

扩张型心肌病 ( DCM)的主要病因是病毒持续感染和自身免疫反应。在 DCM早期可以检测出抗心肌自身抗体 ,抗体介导心肌细胞损伤机制已基本阐明 ,阻止抗心肌抗体介导心肌损伤已成为早期治疗 DCM的有效方法之一。根据分子模仿理论 ,发现 ADP/ ATP载体蛋白与柯萨奇 B3 -病毒外壳蛋白存在同源性 ,酶联免疫吸附试验证实抗 ADP/ ATP载体 2 7- 36肽抗体可以与来源于柯萨奇 B3 病毒 1 2 1 8- 1 2 2 8肽发生免疫反应 ;鼠肝炎病毒、人类巨细胞病毒与 β1-肾上腺素能受体之间具有相同的抗原决定簇 ,还发现抗ADP/ ATP载体抗体可以与钙通道蛋白…     

肠病毒感染及抗ADP/ATP载体抗体在病毒性心脏病发病中的意义

目的 :探讨肠病毒感染与抗二磷酸腺苷 /三磷酸腺苷 (ADP/ ATP)载体抗体在病毒性心脏病发病中的意义。方法 :应用逆转录聚合酶链式反应 (RT- PCR)和免疫转印法对临床诊断为病毒性心肌炎 (VMC)和扩张型心肌病 (DCM)患者进行肠病毒核糖核酸 (RNA)及抗 ADP/ ATP载体抗体的检测。结果 :RT- PCR方法检测肠病毒 RNA,VMC组 (n =74) 4 2例 (56.8% )阳性 ,DCM组 (n=2 6) 1 1例 (42 .3% )阳性。与健康组 (n =30 )比较具有显著性差异 (分别 P <0 .0 1 ,P <0 .0 5) ;肠病毒感染与 DCM患者心脏功能降低有明显相关性。免疫转印法检测 VMC(n =34)和 DCM(n =2 6)患者血清抗 ADP/ ATP载体抗体 ,VMC组 2 3例 (67.6% )阳性 ,DCM组 1 2例 (46.2 % )阳性 ,而健康组均为阴性 ;抗 ADP/ ATP载体抗体和肠病毒 RNA的检出相关 (r=0 .442 )。结论 :肠病毒RNA和抗 ADP/ ATP载体抗体在病毒性心脏病患者中检出具有相关性 ,对临床上出现前驱感染和心脏症状的患者 ,及时进行肠病毒 RNA、CVB- Ig M、抗 ADP/ ATP载体抗体的检测 ,有助于 VMC和 DCM的早期诊断。     

扩张型心肌病抗心肌线粒体ADP/ATP载体自身抗体与Th细胞亚群分析

目的 :探讨扩张型心肌病 (DCM)的体液免疫学发病机制。方法 :借助免疫转印技术及流式细胞仪技术同步分析了 33例 DCM患者和 2 8例正常人的血清中抗心肌线粒体 ADP/ATP载体自身抗体和 Th细胞亚群。结果 :DCM组 Th2 细胞在外周血淋巴细胞中占 (0 .5 3± 0 .2 3) % ,对照组 Th2 细胞占 (0 .2 2± 0 .18) % ,两组比较差异有显著性 (P <0 .0 1) ;DCM组抗心肌线粒体 ADP/ATP载体抗体的检出率 (6 0 .6 1% )明显高于对照组(2 1.43% ) (P <0 .0 1) ;DCM抗心肌线粒体 ADP/ATP载体自身抗体 ,阳性组中 Th2 细胞在外周血淋巴细胞中占 (0 .6 7± 0 .31) % ,阴性组 Th2 细胞占 (0 .2 8± 0 .14) % ,两组比较差异有显著性 (P<0 .0 1)。结论 :DCM中 Th2细胞与抗心肌线粒体 ADP/ATP载体自身抗体的产生有关 ,而且 Th2 细胞和抗心肌线粒体 ADP/ATP载体自身抗体参与了 DCM的病理过程。体液免疫反应的异常在 DCM的发病机制中起着十分重要的作用。     

弓形虫ROP5基因与基因佐剂IL-12对小鼠免疫保护性

目的 观察弓形虫ROP5基因疫苗和基因免疫佐剂IL-12共免疫对昆明小鼠所诱导的免疫保护性。方法 大量制备重组质粒pcROP5和pcmIL-12,以100 μg的剂量同时免疫昆明小鼠,PBS组和pcDNA3.1空质粒组作为对照。用ELISA法测定免疫鼠血清中特异性抗体IgG、IgG亚型的表达水平及细胞因子IFN-γ、IL-4的含量,MTT法测定淋巴细胞转化率,观察小鼠受攻击感染弓形虫后的存活时间。结果 质粒pcROP5和pcmIL-12共免疫小鼠3次后,与pcROP5质粒单独免疫组和对照组相比,IL-12基因佐剂的共免疫提高了免疫鼠血清中特异性抗体IgG及IgG亚型IgG2a的表达水平,提高了细胞因子IFN-γ的含量,增加了淋巴细胞的转化率,但IgG1和IL-4的表达水平变化不大;攻击感染后,质粒pcROP5和pcmIL-12共免疫组小鼠存活时间显著延长。结论 弓形虫ROP5基因与基因佐剂pcmIL-12共免疫能增强对小鼠的免疫保护性。     

扩张型心肌病抗心肌抗体的临床观察及其针对性治疗

目的:探讨扩张型心肌病(DCM)的发病机制,观察针对抗心肌抗体进行免疫学治疗的临床疗效和预后.方法:对2001年1月-2007年12月入院治疗的DCM患者(747例)的病史、诊治过程、随访情况进行回顾性分析.结果:747例中抗心肌抗体阳性者527例(70.55%),随访时间为0.9-7.2年,抗心肌线粒体ADP/ATP载体抗体阳性患者(A组)治疗后平均左室舒张末期内径(LVEDd)为(62.53±9.17)mm,左室射血分数(LVEF)为(37.65±11.15)%;抗β1-肾上腺素能受体抗体(抗β1-受体抗体)阳性患者(B组)治疗后LVEDd (61.35±5.68)mm, LVEF (40.65±12.78)%;抗ADP/ATP载体抗体和抗β1-受体抗体均阳性患者(C组)治疗后LVEDd(61.28±7.72)mm, LVEF(38.35±7.05)%.3组与治疗前比较LVEDd差异均有统计学意义(P<0.05)、NYHA心功能分级均改善Ⅰ～Ⅱ级.结论:自身免疫是DCM的常见致病因素,针对抗ADP/ATP载体抗体使用地尔硫和针对抗β1-受体抗体使用美托洛尔治疗DCM均可明显改善患者心脏功能及心室重构,改善预后.     

微小隐孢子虫重组BCG-CP15/60-23疫苗免疫原性的研究

目的探讨微小隐孢子虫重组BCG-CP15/60-23疫苗(rBCG-CP15/60-23)的免疫原性。方法用rBCG-CP15/60-23免疫BALB/c小鼠,免疫前后取血,用ELISA法检测IgG抗体,末次免疫后2周取淋巴细胞作体外培养,测定培养上清IFN-γ、IL-2和IL-12的水平。设实验组、BCG组、载体组和PBS对照组。结果rBCG-CP15/60-23免疫后2周开始小鼠IgG抗体水平逐渐升高,与BCG组和载体组比较差异均有统计学意义(P均<0.01);免疫鼠脾淋巴细胞培养上清中IFN-γ和IL-2的含量与PBS对照组和BCG组比较差异有统计学意义(P均<0.05),IL-12水平组间差异无统计学意义(P>0.05)。结论rBCG-CP15/60-23免疫小鼠后能刺激机体产生IgG抗体,IFN-γ和IL-2的分泌增加,具有较好的免疫原性。     

重组幽门螺杆菌疫苗免疫保护机制的研究

目的 研究以减毒鼠伤寒沙门菌为载体构建的重组幽门螺杆菌（Hp)疫苗诱导小鼠产生保护性免疫应答的机制。方法 将表达Hp尿素酶B亚单位（UreB)，黏附素（HpaA)及尿素酶B亚单位/黏附素融合蛋白（UreB/HpaA)的减毒鼠伤寒沙门菌（Salmonella typhimurium)给小鼠分别灌胃，另设单纯减毒鼠伤寒沙门菌和生理盐水免疫鼠为对照，免疫4周后以Hp活菌攻击，观察各组小鼠的免疫保护率，攻击前后血清中抗Hp抗体IgC1,IgG2a和IgA的变化。小鼠脾脏和胃黏膜中γ干扰素（IFN-γ）和白介素-4（IL-4）mRNA表达变化。结果 UreB,HpaA及UreB/HpaA组的免疫保护率分别为50%，41%和77%，和生理盐水组相比，攻击前各鼠伤寒沙门菌免疫组IgG1,IgG2a均轻度升高而IgA无变化，攻击后各鼠伤寒沙门菌免疫组IgG2a升高显著并以UreB/HpaA组为最，而IgG1和IgA的升高无统计学差异。胃黏膜攻击前生理盐水组无IFN-γ表达，其余各组均100%表达；攻击后生理盐水组IFN-γ轻度表达，但仍明显低于各鼠伤寒沙门菌免疫组，IL-4在攻击前后各组均无表达，脾IFN-γ和IL-4在所有组攻击前后均全部表达。结论 以减毒鼠伤寒沙门菌为载体构建的Hp疫苗在小鼠体内诱导出以TH1反应为主的保护性免疫应答。     

多房棘球绦虫elp基因核酸疫苗诱生免疫应答的实验研究

目的观察多房棘球绦虫elp基因核酸疫苗免疫BALB/c小鼠引起的体液和细胞免疫应答。方法30只BALB/c小鼠随机分成3组:Ⅰ组,肌注空质粒(pcDNA3.1)100μg/鼠;Ⅱ组,肌注多房棘球绦虫elp基因真核表达重组质粒(pcD-ELP)100μg/鼠;NS组,肌注等体积生理盐水。每2周免疫1次,共3次。ELISA方法检测免疫后不同时间产生的特异性IgG1,IgG2a和IgG 2 b水平。双抗体夹心ELISA法检测免疫鼠脾脏单个核细胞(PMNC)在受到特异抗原刺激后,产生IL-4I、L-12和IFN-γ的能力,3H-TdR掺入法检测脾淋巴细胞的增值能力。结果首次免疫后4周,Ⅱ组小鼠血清可检测到特异性IgG1I、gG2a和IgG2b抗体,随免疫时间和免疫次数的增加3种特异性抗体的OD值逐渐增高,实验结束时(首次免疫后7周)3种特异性抗体OD值均最高。Ⅰ组和Ⅱ组小鼠脾PMNC培养上清中IFN-γ浓度分别为50 pg/ml和55 pg/ml,受特异抗原刺激后分别为44 pg/ml和101.5 pg/ml。NS组IFN-γ及各组IL-4和IL-12均未检出。Ⅰ组和Ⅱ组小鼠脾淋巴细胞增殖能力增高,其淋巴细胞CPM值分别为NS组的85倍和123倍,受到特异抗原或刀豆素刺激其淋巴细胞增殖更明显。结论多房棘球绦虫elp基因核酸疫苗可刺激BALB/c小鼠同时产生很强的细胞免疫和体液免疫应答。     

结核分枝杆菌Rv0033的原核表达及其免疫原性研究

目的表达重组结核分枝杆菌(Mycobacterium tuberculosis,M.tb)Rv0033,并观察其免疫原性,为结核病(Tuberculosis, TB)新型疫苗的研发筛选优势靶抗原。方法构建重组载体pET30b-Rv0033并表达重组Rv0033。以全血干扰素释放技术(Whole blood IFN-γrelease assay, WBIA)检测M.tb感染者的T细胞是否特异性识别重组载体pET30b-Rv0033。取纯化的rRv0033混合佐剂DMT后免疫小鼠,ELISA法检测血清特异性抗体IgG、IgG1及IgG2a水平,并计算IgG2a/IgG1比值;同时检测小鼠脾细胞中特异性IFN-γ、TNF-α、IL-2水平。结果成功构建原核表达载体pET30b-Rv0033,重组Rv0033的诱导表达、纯化和鉴定结果均符合预期。人群外周血淋巴细胞经rRv0033蛋白刺激后,非典型结核(Atypical tuberculosis, ATB)及潜伏性结核感染(Latent tuberculosis infection, LTBI)受试者IFN-γ水平显著高于健康对照人群(P0.01);ATB受试者IFN-γ水平显著高于LTBI受试者(P0.05)。BCG+Rv0033/DMT免疫小鼠产生的特异性抗体(IgG、IgG1和IgG2a)滴度水平显著高于Rv0033/DMT组和BCG组(均P0.01);Rv0033/DMT组和BCG+Rv0033/DMT组小鼠的IgG2a/IgG1比值显著高于DMT组和BCG组,且1,提示rRv0033可诱导以Th1细胞免疫应答为主的免疫应答。接受PPD刺激或rRv0033蛋白刺激的BCG+Rv0033/DMT组小鼠IFN-γ、TNF-α及IL-2水平较高,其次分别是BCG组、Rv0033/DMT组、DMT组,PBS组水平较低,且rRv0033蛋白混合DMT佐剂组高于单纯DMT佐剂组(P0.05)。结论制备的rRv0033蛋白可被M.tb感染者外周血T细胞特异性识别,联合佐剂DMT免疫小鼠能够诱导高水平的抗原特异性Th1型免疫应答,可能与rRv0033蛋白提供的免疫保护力密切相关。     

Metabolic remission with octreotide in patients with insulinoma

Abstract. Objectives . The efficacy of octreotide was studied in a group of patients with biochemical evidence of insulinoma. Design . A phase-II study Setting . A university department of internal medicine. Subjects . Seven patients with biochemical evidence of insulinoma and without metastatic lesions. Intervention . Daily treatment with octreotide, a somatostatin analogue, mainly within the dosage of 100–300 μg day^-1. The treatment was continued in patients with biochemical evidence of response or until surgery was undertaken. Main outcome . Five patients avoided hypoglycaemic symptoms and had normalization of blood glucose values for a median of 15+ months (range 0.2–54 months). Two did not improve metabolically. The treatment was well tolerated and had no deleterious effects on blood glucose regulation. Conclusion . Octreotide seems to be a promising treatment for many of the patients with insulinoma who are not suitable for surgery.     

Changes in coagulation parameters with exercise in patients with classic hemophilia

Vigorous exercise is known to increase VIII:C and VIIIR:Ag levels transiently in normal individuals. Although exercise programs are frequently advocated in the management of hemophilia, the effects of exercise on coagulation parameters in these patients have not been well studied. Eleven hemophiliacs were exercised on a bicycle ergometer to maximum voluntary effort as evidenced by an increase in pulse, blood pressure, and plasma catecholamine (norepinephrine and epinephrine) levels. The effects of this exercise on coagulation parameters, including functional and antigenic components of the factor VIII molecule, were determined. The entire group demonstrated a decrease in mean prothrombin time (11.7 to 11.2 sec). Four mild hemophiliacs demonstrated an increase in mean VIII:C (14.5% to 17.3%), and VIII:CAg (12% to 17.8%). Changes in VIII:C and VIII:CAg were not noted in the seven severe hemophiliacs. Both severe and mild patients demonstrated significant changes in fibrinogen, factor II, and factor VII after exercise. This study indicates that submaximal exercise modifies coagulation parameters in patients with hemophilia.     

Experience with ponatinib in paediatric patients with leukaemia

Ponatinib has proven to be effective in adults with Philadelphia chromosome-positive leukaemias, but data in paediatrics are scarce. Among paediatric patients with chronic myeloid leukaemia (n = 9) or acute lymphoblastic leukaemia (n = 12) treated with varying doses of ponatinib in 13 centres, 71% showed a decrease in disease burden after a median of three months. Ponatinib was well tolerated, with grade 3 toxicities occurring in 29% of patients. Toxicities were similar to those reported in adults, with the exception of arterial thrombotic events, which were not observed. Ponatinib has a favourable safety profile in this paediatric cohort, but dose-finding studies are needed.     

Mental disorders in patients with obesity in comparison with healthy probands

OBJECTIVE: Findings concerning the association of obesity and mental disorders are inconsistent. The present epidemiological study investigates adjusted 4-week, 12-month, and lifetime prevalence rates of mental disorders in obese individuals compared with physically healthy probands and overweight individuals. Correlates of the associations are examined. METHODS: Prevalence rates were calculated from two large epidemiological surveys from both the general population of Germany and inpatient centers. The surveys investigated subjects with obesity (n=910) and overweight (n=1550), as well as physically healthy probands (n=495). The prevalence rates were based on the Munich-composite international diagnostic interview, a standardized interview for the assessment of mental disorders. Correlates of mental disorders in obese individuals were assessed using self-report questionnaires and medical examinations. RESULTS: The adjusted odds ratios (OR) of obese inpatients and obese patients from the general population were significantly elevated in comparison with healthy probands for the 4-week (OR: 2.2; 2.3), 12-month (OR: 1.8; 2.7) and lifetime (OR: 1.4; 2.0) periods. Prevalence rates of overweight individuals were below those of obese individuals. Mood, anxiety and somatoform disorders were most frequent. In particular, sex, marital status and comorbid musculoskeletal diseases proved to be correlates of an increased risk for mental disorders in obese individuals. The presence of comorbid mental disorders was associated with significantly increased health care use and lower quality of life. CONCLUSIONS: There is a strong relationship between obesity and mental disorders. A future task is to improve care of mental disorders in patients with obesity.     

Therapy with budesonide in patients with refractory sprue

INTRODUCTION: Refractory sprue (RS) is a rare malabsorption syndrome, which often requires long-term corticosteroid treatment. Locally acting budesonide could replace systemic corticosteroid therapy and reduce toxicity in patients with RS. Aims: To evaluate the efficacy and toxicity of budesonide in patients with RS. PATIENTS AND METHODS: Clinical and histological data from patients with RS who received budesonide were analyzed. RS was defined as villous atrophy and malabsorption in spite of a strict gluten-free diet persisting for >6 months or requiring earlier therapeutic intervention. RESULTS: We identified 9 patients (1 with autoimmune enteropathy, 4 with RS type I without and 3 with RS type II with signs of early T cell lymphoma and 1 with CD4-positive sprue-like intestinal T cell lymphoma), who received 9 mg/day of budesonide (range 6-12) for 24 months (1-60), and 7 of whom had an initial treatment with 40 mg/day of prednisolone (30-60) for 4 months (1-144). The initial body mass index was 18 (13.1-22.8) and increased similarly under prednisolone [21.5 (14.9-26.7), p < 0.05] and budesonide therapy [21 (18-27.2), p < 0.05]. The stool frequency per day also decreased similarly from 6 (2-8) to 2 (1-3) and 2 (1-5), each p < 0.05, under prednisolone and budesonide therapy, respectively. Two patients with RS type II did not respond and 7, including all 4 with RS type I, were clinically stable with budesonide therapy. Skin fragility in 1 patient was the only adverse effect of budesonide therapy. CONCLUSIONS: Budesonide may be an effective treatment option in patients with RS type I, which can stabilize the clinical condition similar to prednisolone.     

Gammagraphy with sucralfate labelled with technetium in esophagitis

In previous studies it has been reported that, after being labeled with technetium, sucralfate, an useful drug in peptic diseases, can be used to detect peptic lesions of the digestive tract. In this work we report our experience with this technique in the diagnosis of esophagitis. 25 studies (11 controls and 14 patients) were undertaken. Sucralfate scintigraphy was normal in the 11 control studies, and abnormal in 10 out of 14 patients. Scintigraphy was abnormal in peptic as well as caustic lesions.     

Polymyositis-like syndrome with rhabdomyolysis in association with brucellosis

Diffuse myositis with progression to rhabdomyolysis has been reported in association with wide range of viral infections. We report a case of polymyositis-like syndrome complicated by rhabdomyolysis secondary to brucellosis. This case report thus contributes yet another atypical presentation to a disease already infamous for its protean manifestations.