McAb4F8,McAb1F11与囊尾蚴抗原作用免疫印渍分析

将猪囊尾蚴囊液抗原（ＣＣＦＡ）和脑囊虫病病人脑脊液（ＰＣＳＦ）进行ＳＤＳ－ＰＡＧＥ电泳分析，发现在ＣＣＦＡ中有１４条蛋白带，范围在２８～７６ｋＤａ。脑囊虫病病人ＣＳＦ中有６条蛋白整，范围在３３～８３ｋＤａ。用ＣＣＦＡ和ＰＣＳＦ与ＭｃＡｂ４Ｆ     

肝脏特异的F蛋白(F Protein)

Fravi 等在1968年从肝组织中发现人体肝脏特异的 F 抗原(Fantigen),1971年 Rosen-mud 在几例肝病患者血清中检测出肝脏特异的自身抗原—F 抗原以来,国外学者对 F 抗原的认识逐步深入,证实它是肝细胞特有的蛋白质,但对其功能尚不十分清楚。F 蛋白定位于肝细胞浆内,具有高度组织特异性(只存在于肝细胞内)、种属一致性(大鼠、小鼠、豚鼠、兔、牛、羊和人类同),肝细胞内 F 蛋白含量甚高,而血液中含量甚微,含量之比为40000:1。血液中微量的 F 蛋白来源于肝细胞损伤或正常代谢,以致细胞破坏而释放的结果。J.B.Smith 等发现 F 抗原出现在人体血清中,主要     

Pulmonalvenenisolation zur Behandlung von Vorhofflimmern. Für wen, welche Methode, mit welchen Ergebnissen?

Atrial fibrillation is a relevant challenge for health care systems in Europe and North America. Interventional treatment of atrial fibrillation in terms of ablation of atrial structures including isolation of the pulmonary veins is nowadays established as an alternative to drug therapy, especially if treatment with antiarrhythmic drugs has been ineffective. Recently published European and North American guidelines support this strategy.This article describes different techniques of atrial fibrillation ablation including intracardiac echo-guided pulmonary vein isolation. Potential complications, success rates and different follow-up strategies are specified. Furthermore interventional treatment of atrial fibrillation of heart failure patients is discussed.     

Molecular phenotypes of human parvovirus B19 in patients with myocarditis

AIM:To investigate molecular phenotypes of myocardial B19V-infection to determine the role of B19V in myocarditis and dilated cardiomyopathy(DCM).METHODS:Endomyocardial biopsies(EMBs) from 498 B19V-positive patients with myocarditis and DCMwere analyzed using molecular methods and functional experiments.EMBs were obtained from the University Hospitals of Greifswald and Tuebingen and additionally from 36 German cardiology centers.Control tissues were obtained at autopsy from 34 victims of accidents,crime or suicide.Identification of mononuclear cell infiltrates in EMBs was performed using immunohistological staining.Anti-B19V-IgM and anti-B19V-IgG were analyzed by enzyme-linked immunosorbent assay(ELISA).B19V viral loads were determined using in-house quantitative real-time polymerase chain reaction(PCR).For B19V-genotyping a new B19V-genotype-specific restriction fragment length polymorphism(RFLP)-PCR was established.B19V-genotyping was verified by direct DNAsequencing and sequences were aligned using BLAST and BioEdit software.B19V P6-promoter and HHV6-U94-transactivator constructs were generated for cell culture experiments.Transfection experiments were conducted using human endothelial cells 1.Luciferase reporter assays were performed to determine B19Vreplication activity.Statistical analysis and graphical representation were calculated using SPSS and Prism5 software.RESULTS:The prevalence of B19V was significantly more likely to be associated with inflammatory cardiomyopathy(iCMP) compared to uninflamed DCM(59.6% vs 35.3%)(P 0.0001).The detection of B19V-mRNA replication intermediates proved that replication of B19V was present.RFLP-PCR assays showed that B19V-genotype 1(57.4%) and B19V-genotype 2(36.7%) were the most prevalent viral genotypes.B19V-genotype 2 was observed more frequently in EMBs with iCMP(65.0%) compared to DCM(35%)(P = 0.049).Although there was no significant difference in gender-specific B19V-loads,women were more frequently infected with B19V-genotype 2(44.6%) than men(36.0%)(P = 0.0448).Coinfection with B19V and other cardiotropic viruses was found in 19.2% of tissuesamples and was associated with higher B19V viral load compared to B19V-monoinfected tissue(P = 0.0012).The most frequent coinfecting virus was human herpes virus 6(HHV6,16.5%).B19V-coinfection with HHV6 showed higher B19V-loads compared to B19V-monoinfected EMBs(P = 0.0033),suggesting that HHV6 had transactivated B19V.In vitro experiments confirmed a 2.4-fold increased B19V P6-promoter activity by the HHV6 U94-transactivator.CONCLUSION:The finding of significantly increased B19V loads in patients with histologically proven cardiac inflammation suggests a crucial role of B19V-genotypes and reactivation of B19V-infection by HHV6-coinfection in B19V-associated iCMP.Our findings suggest that B19V-infection of the human heart can be a causative event for the development of an endothelial cell-mediated inflammatory disease and that this is related to both viral load and genotype.     

Relevance of cardiac parvovirus B19 in myocarditis and dilated cardiomyopathy: review of the literature

Over the last decade, parvovirus B19 (B19V) has frequently been linked to the pathogenesis of myocarditis (MC) and its progression towards dilated cardiomyopathy (DCM). The exact role of the presence of B19V and its load remains controversial, as this virus is also found in the heart of healthy subjects. Moreover, the prognostic relevance of B19V prevalence in endomyocardial biopsies still remains unclear. As a result, it is unclear whether the presence of B19V should be treated. This review provides an overview of recent literature investigating the presence of B19V and its pathophysiological relevance in MC and DCM, as well as in normal hearts. In brief, no difference in B19V prevalence is observed between MC/DCM and healthy control hearts. Therefore, the question remains open whether and how cardiac B19V may be of pathogenetic importance. Findings suggest that B19V is aetiologically relevant either in the presence of other cardiotropic viruses, or when B19V load is high and/or actively replicating, which both may maintain myocardial (low‐grade) inflammation. Therefore, future studies should focus on the prognostic relevance of the viral load, replicative status and virus co‐infections. In addition, the immunogenetic background of MC/DCM patients that makes them susceptible to develop heart failure upon presence of B19V should be more thoroughly investigated.     

人细小病毒B19感染在心肌病发病中的作用

目的 :探讨人细小病毒B19(HPVB19)感染在心肌病发病中的作用。方法 :病理诊断确诊为炎症性心肌病患者 30例 ,扩张型心肌病 (DCM)患者 36例 ,肥厚型心肌病 (HCM)患者 13例 ,无心肌炎、DCM或近期感染者 (对照 ) 36例。所有被研究者均行心肌组织活检 ,用于病理组织学检查和巢式聚合酶链反应 (PCR)检测分析HPVB19 DNA。结果 :炎症性心肌病中有 4例HPVB19 DNA阳性 (占 13.3% ) ,DCM中有 3例HPVB19 DNA阳性 (占 8.3% ) ,HCM中有 2例HPVB19 DNA阳性 (占 15 .4 % ) ,而对照组HPVB19 DNA均为阴性。结论 :心肌病患者中HPVB19感染率较高 ,HPVB19感染可能是心肌病的重要病因之一。     

Possible relationship between acute viral myocarditis and dilated cardiomyopathy]

To explore the possible relationship between acute viral myocarditis and dilated cardiomyopathy (DCM), 35 acute diffuse viral myocarditis (ADVM) patients with cardiac enlargement were studied for 6 years on average. The results showed that: (1) In 22 ADVM patients, the dilated hearts had returned to normal on X-ray films. The other 13 cases still had cardiac enlargement complicated with various degrees of cardiac insufficiency (NYHA II/III) and ECG abnormalities. The manifestation of these 13 patients resembled those seen in the early stage of DCM. (2) Serum neutralizing antibody titres of Coxsackie B virus (CBV) in 35 ADVM patients after 6 years observation on average were significantly higher than those cases with cardiac enlargement induced by other causes (P less than 0.01). High neutralizing antibody titres (greater than or equal to 320) to CBV were more common among the patients with ADVM 65.7% vs 25.7% (P less than 0.05). The results indicate that some cases of ADVM might develop into DCM.     

肌球蛋白致自身免疫性心肌疾病的实验研究

目的　探索在没有病毒感染的情况下 ,自身免疫机制在心肌炎和心肌病发病中的作用。方法　 (1)猪心肌肌球蛋白致BALB/C鼠自身免疫性心肌疾病模型的复制 :实验组 (n =32 )于实验的第 0 ,7和 30天 3次注射肌球蛋白 ,对照组 (n =2 0 )与实验组同样的时间注射弗氏完全佐剂 ,于初次免疫后的第 15 ,2 1和 12 0天分别留取血清和心肌标本。 (2 )病毒性心肌炎模型的复制 :病毒接种组 (n =2 2 )给予 10 3 TCID50 / 0 1mlCVB3 溶液腹腔注射 ,对照组 (n =10 )给予Vero细胞冻融液 0 1ml腹腔注射 ,接种时间及采样时间同肌球蛋白免疫组。 (3)抗体检测 :ELISA法检测小鼠血清抗肌球蛋白抗体。结果　病理结果显示 ,肌球蛋白致心肌炎小鼠急性期 (15和 2 1d)以心肌纤维变性坏死和淋巴细胞浸润较明显 ,间质病变较轻 ,内膜和心瓣膜无明显改变 ;慢性期 (12 0d)仅见心肌纤维化 ,急慢性期均可检测到高滴度的抗肌球蛋白抗体 (P <0 0 0 1) ,对照组为正常心肌 ,抗体滴度低。病毒接种组慢性期亦显示成纤维细胞增生。结论　心肌肌球蛋白可以作为自身抗原刺激机体产生自身免疫反应 ,在没有病毒感染的情况下 ,单一的自身免疫机制即可导致心肌炎向心肌病的转化。     

Parvovirus B19 genotypes 1 and 2 detection with real-time polymerase chain reaction assays

BACKGROUND AND OBJECTIVES: Parvovirus B19 (B19V) DNA screening has been introduced to comply with European regulations for certain plasma products. Current commercial and some in-house B19V DNA assays fail to detect or under-quantify the recently identified genotypes 2 and 3. In this report, we describe 2-year experience with B19V DNA screening using the commercial assay from Roche (detecting only genotype 1) combined with an in-house assay (detecting genotypes 1, 2 and 3). This dual testing approach enables the identification of molecular variants of B19V. MATERIALS AND METHODS: Between 2005 and 2007, approximately 2.6 million plasma donations were screened for B19V DNA loads exceeding 10(6) IU/ml using the Roche and the in-house real-time polymerase chain reaction assay. RESULTS: A total of 232 plasma units were identified with B19V DNA loads above 10(6) IU/ml. Concordant results were observed for the majority of B19V positive samples; however, three of these showed discrepant results between the two assay systems. One was a B19V genotype 2 strain not detected by the Roche assay; another was a B19V genotype 1 strain with a mismatch in the 3'-end of the reverse primer and therefore under-quantified by the Roche assay; and the third one was also a B19V genotype 1 strain that gave an unusual amplification plot in the in-house assay due to a mismatch in the probe-binding site. CONCLUSIONS: New, high viral load, B19V genotypes 2 and 3 infections are rare in blood donors tested by Sanquin. One case was found while testing 2.6 million donations. The prevalence of B19V genotype 1 variants not detected by commercial or in-house assays might be in the same range or even higher than the prevalence of B19V genotype 2 viruses, which remain undetected.     

Angioimmunoblastic T-cell lymphoma: histological progression associates with EBV and HHV6B viral load

The clinical and histological presentations of angioimmunoblastic T-cell lymphoma (AITL) often mimic an infectious process. Epstein-Barr virus (EBV) and human herpes virus (HHV6) are known to be associated with AITL, but whether these viral infections play a role in its pathogenesis is unclear. It also remains to be investigated whether there might be other viruses associated with AITL. We first screened 26 well-characterised cases of AITL for herpesvirus by polymerase chain reaction (PCR) with universal primers and found evidence of only EBV and HHV6B infection. Subsequent PCR using virus-specific primers demonstrated EBV and HHV6B infection in 40/49 biopsies (36/42 cases) and 21/49 biopsies (19/42 cases) of AITL respectively with both viral infections found in 17/49 specimens (15/42 cases). Importantly, simultaneous infection with both viruses was found only in specimens showing histological pattern II (n = 2) or III (n = 15). Interestingly, among specimens containing both viruses, there was a tendency towards an inverse correlation between the EBV and HHV6B viral load as shown by quantitative PCR. In specimens positive only for EBV, the viral load was significantly higher in specimens with histological pattern III than those with pattern II. High EBV load was also significantly associated with B-cell monoclonality. Double EBV encoded small RNA (EBER) in situ hybridisation and immunohistochemistry indicated that EBV-infected B cells had a late postgerminal centre immunophenotype. Our results demonstrate an association between EBV and HHV6B infection and the histological progression of AITL, suggesting that these viruses may play a role in the pathogenesis of this lymphoma.     

Description of a local cardiac adiponectin system and its deregulation in dilated cardiomyopathy.

AIMS: Despite recent advances in medical therapy, heart failure remains a leading cause for cardiovascular mortality, and its complex pathogenesis is incompletely understood. This study was performed to identify possible new therapeutic targets in dilated cardiomyopathy (DCM). METHODS AND RESULTS: Oligonucleotide microarray analysis was performed on endomyocardial biopsies (EMBs) from patients with early DCM (LVEDD > or = 55 mm, LVEF < or = 55%, n = 5) and control subjects (LVEDD < 55 mm, LVEF > 60%, no cardiac pathology, n = 4). Adiponectin, an adipocytokine involved in cellular metabolism, survival, and immunmodulation, was six-fold downregulated in DCM patients. Microarray data for adiponectin were confirmed by TaqMan-PCR (9.2-fold downregulation, control n= 9 vs. DCM n= 9, respectively, P < 0.05). Immunohistological analysis of EMBs showed significant downregulation of cardiac adiponectin protein expression independent of serum adiponectin (P = 0.36, ns) or serum TNFalpha concentrations (P = 0.46, ns). Neither the adiponectin receptor 1 (adipo-R1) nor adipo-R2 was deregulated in early DCM. Adiponectin mRNA and protein downregulation were confirmed in explanted hearts of patients with advanced DCM (LVEF < 25%, n= 8). In vitro, adiponectin incubation of neonatal rat ventricular myocytes led to activation of the pro-survival kinase PKB/Akt, increased eNOS-phosphorylation, and prevented stress-induced apoptosis of cardiomyocytes in an Akt-dependent manner. Moreover, inhibition of adiponectin secretion was accompanied by an increase in the expression of the cytokine and its receptors. CONCLUSION: These data indicate the existence of a local cardiac adiponectin system regulated independent of adiponectin and TNFalpha serum levels and its disturbance in cardiac pathology. The study suggests a role for adiponectin in the pathogenesis of DCM and implicates the adipocytokine as a possible future therapeutic target in DCM.