Role natural killer group 2D-ligand interactions in hepatitis B infection

Hepatitis B virus （HBV） infection is the leading causeof liver disease and hepatocellular carcinoma （HCC）worldwide, in spite of prophylactic vaccination andantiviral treatment modalities. The immunopathogenesisof HBV infection has been intensively studied and ispropelled by complex interactions between the virus andthe host immune system. Natural killer group 2D （NKG2D）is a well-characterized activating receptor, expressed onnatural killer （NK） cells, NK T cells and CD8＋ cytotoxic Tcells. This receptor is present in both humans and miceand binds to a diverge family of ligands that resemble theMHC-class Ⅰ molecules. Increasing evidence shows thatNKG2D-ligand interactions are critical in the establishmentof HBV persistence and the development of liver injuryand HCC. The expression of NKG2D ligands dependson the presence of several polymorphisms and is alsomodulated post-transcriptionally by HBV. While it isknown that HBV circumvents host’s innate immunityvia the NKG2D pathway but the exact mechanismsinvolved are still elusive. This letter discusses previousaccomplishments on the role of NKG2D ligand regulationin the development of chronic HBV, liver injury and HCC.Key words： Hepatitis B virus; Natural killer group 2Dreceptor; Natural killer cells; MHC class I polypeptiderelatedchain A; Hepatocellular carcinoma

Circulating Adhesion Molecules in Patients with Keshan Disease and Their Relationship with Coxsackie B Virus Infection

This study determined the levels of serum soluble intercellular adhesion molecule-1 （sI-CAM-l） and soluble vascular cell adhesion molecular-1 （sVCAM-1） in patients with different types of Keshan disease （KD）, examined the relationship between Coxsackie B virus-specific IgM antibody （CBV-IgM） and slCAM-1 or sVCAM-1 in KD patients, and investigated the role of these adhesion molecules in the pathogenesis of KD and their clinical implications. The levels of serum slCAM-1, sVCAM-1 and CBV-IgM were measured by using enzyme-linked immunosorbent assay in 22 patients with chronic Keshan disease （CKD）, 27 with latent Keshan disease （LKD） and 28 healthy controis. The subjects in different groups were adjusted for sex and age. Echocardiography was adopted to determine left ventricular ejection fraction （LVEF） in 22 patients with CKD. The results showed that CKD patients had significantly higher levels of slCAM-1 and sVCAM-1 than LKD patients and healthy controls （P〈0.01 for all）. And there was significant difference in the levels of the 2 adhesion molecules between LKD patients and healthy controls （P〈0.05）. A negative correlation was found between LVEF and slCAM-1 or sVCAM-1 in CKD patients. The percentage of CBV-specific IgM positive individuals in KD patients was significantly higher than that of healthy controls. In CVB-specific IgM positive patients, the levels of serum slCAM-1 and sVCAM-1 were significantly greater than those in CBV-specific IgM negative counterpart. It was concluded that the increase in the levels of slCAM-1 and sVCAM-1 suggests the progression of inflammation in KD. slCAM-1 and sVCAM-1 can promote the development of myocardial pathology and lead to poor myocardial function. The increased serum slCAM-1 and sVCAM-1 in KD patients may be related to CBV infection.

Design,synthesis and biological activity of some novel benzimidazole derivatives against Coxsackie virus B_3

A series of novel benzimidazole derivatives was synthesized and their anti-Coxsackie virus B3 （CVB3） activity was evaluated in VERO ceils. Compounds 9 and 10 exhibited better inhibitory activity than those of ribavirin （RBV） with IC50 values of 5.30 and 1.06 μg/mL, respectively. ？2009 Xian Jin Luo. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Design, Synthesis and Inhibitory Properties against Coxsackie B3/B6 of Some Novel Triazole Derivatives

A series of 1,2,4-triazole derivatives were synthesized, and their abilities to inhibit the in vitro replication of Coxsackie B3/B6 were evaluated. Among the 1,2,4-triazole derivatives, compound 3 g displayed potent activity, with a high antiviral potency (IC50 = 1.71 μM (against CVB3), 1.43 μM (against CVB6)). The structures of all the new synthesized compounds were confirmed by 1H-NMR spectra, mass spectra and elemental analyses.

Lymphocyte-to-white blood cell ratio is associated with outcome in patients with hepatitis B virus-related acute-on-chronic liver failure

BACKGROUND The lymphocyte-to-white blood cell ratio(LWR)is a blood marker of the systemic inflammatory response.The prognostic value of LWR in patients with hepatitis B virus-associated acute-on-chronic liver failure(HBV-ACLF)remains unclear.AIM To explore whether LWR could stratify the risk of poor outcomes in HBV-ACLF patients.METHODS This study was conducted by recruiting 330 patients with HBV-ACLF at the Department of Gastroenterology in a large tertiary hospital.Patients were divided into survivor and non-survivor groups according to their 28-d prognosis.The independent risk factors for 28-d mortality were calculated by univariate and multivariate Cox regression analyses.Patients were divided into low-and high-LWR groups according to the cutoff values.Kaplan-Meier analysis was performed according to the level of LWR.RESULTS During the 28-d follow-up time,135 patients died,and the mortality rate was 40.90%.The LWR level in non-surviving patients was significantly decreased compared to that in surviving patients.A lower LWR level was an independent risk factor for poor 28-d outcomes(hazard ratio=0.052,95%confidence interval:0.005-0.535).The LWR level was significantly negatively correlated with the Child-Turcotte-Pugh,model for end-stage liver disease,and Chinese Group on the Study of Severe Hepatitis B-ACLF II scores.In addition,the 28-d mortality was higher for patients with LWR<0.11 than for those with LWR≥0.11.CONCLUSION LWR may serve as a simple and useful tool for stratifying the risk of poor 28-d outcomes in HBVACLF patients.

柯萨奇病毒B组5型非结构蛋白抑制NF-κB信号通路的作用机制研究

目的 柯萨奇病毒B组5型(CVB5)是手足口病的重要病原体之一,可导致发热、皮疹或疱疹等临床症状,重症者出现神经系统疾病,甚至死亡。天然免疫应答是机体抗病毒入侵的第一道防线,其中核因子κB (NF-κB)是宿主天然免疫反应中的重要蛋白质,然而关于CVB5感染后调控NF-κB介导信号通路的研究尚鲜有报道。方法 本研究通过检测启动子活性、促炎因子水平以及通路中关键蛋白表达等,阐明CVB5对NF-κB信号通路的调控作用机制。结果 CVB5感染可抑制促炎因子表达和p65的磷酸化。CVB5非结构蛋白(NSP)可抑制促炎因子表达以及重要蛋白p65和IκBα的磷酸化。经STRING11.1数据库预测表明,CVB5 3CD蛋白与宿主多聚胞嘧啶结合蛋白1 (PCBP1)具有相互作用,且PCBP1可促进IκBα和p65的磷酸化,抑制病毒复制。结论 CVB5 NSP可负调控NF-κB信号通路,且与3CD相互作用的PCBP1蛋白可通过调控NF-κB通路抑制CVB5复制。本研究探索病毒与宿主天然免疫应答的调控作用,从而为研制抗CVB5感染的药物提供作用靶点。

黄芪皂甙对小鼠柯萨奇B_3病毒性心肌炎的治疗作用

目的：探讨黄芪皂甙对 Ｂ Ａ Ｌ Ｂ／ｃ 小鼠柯萨奇 Ｂ３ （ Ｃ Ｖ Ｂ３ ）病毒性心肌炎的治疗作用。方法：雄性 Ｂ Ａ Ｌ Ｂ／ｃ 小鼠腹腔接种 Ｃ Ｖ Ｂ３ ，制成急性病毒性心肌炎模型，接种１ ｈ 后腹腔内注入黄芪皂甙 ５．６４×１０－ ３ ｍ ｇ／ｇ，１ 次／ｄ ×７ ｄ，观测小鼠存活率、血浆乳酸脱氢酶（ Ｌ Ｄ Ｈ）和谷草转氨酶（ Ｇ Ｏ Ｔ）、体温、心肌病理变化和心肌病毒滴度的影响。结果：黄芪皂甙治疗组比病毒感染组存活率明显增高，血浆 Ｌ Ｄ Ｈ 和 Ｇ Ｏ Ｔ 明显降低，体温保持正常，心脏质量／体质量值显著降低，心肌病理变化减轻，心肌匀浆病毒滴度降低（ Ｐ＜ ０．０５）。结论：黄芪皂甙有治疗病毒性心肌炎的作用。

应用VP1蛋白的合成肽抗原检测心肌炎患者血清中柯萨奇B病毒IgM抗体

从柯萨奇B病毒(CVB)衣壳蛋白VPI的保守区选出一段13肽进行化学合成,经ELISA证买该段多肽是CVB的共同优势抗原表位.以多肽作包被抗原,建立了检测CVBIgM的间接ELISA法,对85例健康人、230例病毒性心肌炎、19例RF阳性血清、40例其它疾病患者进行了检测,阳性率分别为7.1%、41.3%、21.1%、10%.230例心肌炎中有106例用以病毒作包被抗原的间接ELISA进行了CVBIgM检测,阳性率49.1%.两者之间结果有良好的相关性(X^2=41.9,P<O.0001),阳性率无明显差别(X^2=2.45,P>0.05).提示,应用合成肽抗原检测CVBIgM抗体有较好的敏感性和特异性.

柯萨奇B组病毒3种检测方法的比较

目的探讨限制性内切酶分析法检测柯萨奇B组病毒的特异性和重现性。方法收集500例疑似病毒性心肌炎的患者血清,分别采用限制性内切酶分析法、ELISA法、RT-PCR法3种检测方法进行检测,每种方法在相同条件下连续重复3次,然后将其结果进行比较。结果ELISA法的3次检测的结果差别较大,阳性率分别为55.4%、58.4%、60.6%;RT-PCR法的3次检测的结果差别相对较小,阳性率分别为51.1%、51.6%、52.8%;限制性内切酶分析法的3次检测结果完全一致。结论限制性内切酶分析法是一种特异性强、重现性好,明显优于ELISA法和RT-PCR法的新的病毒检测方法。

新生儿柯萨奇B＿3病毒性心肌炎及其病原学检测

１９９２年北京市某医院发生一次新生儿柯萨奇Ｂ３病毒感染流行，发病３５例，合并病毒性心肌炎８例，２例死亡，尸解病理诊断为急性弥漫性非化脓性心肌炎。为明确病原学诊断，用ＥＬＩＳＡ测患儿血中柯萨奇Ｂ病毒抗体ＩｇＭ；用ＰＣＲ测患儿血及心肌组织中的肠道病毒（包括柯萨奇病毒）ＲＮＡ；用大便及咽拭子分泌物进行病毒分离，证实此次流行的病原为柯萨奇病毒Ｂ３。认为ＥＬＩＳＡ、ＰＣＲ方法快速、敏感、特异强，可推广应用。

实验性小鼠柯萨奇B_3病毒心肌炎自然杀伤细胞活性和干扰素测定

测定了BALB/c鼠柯萨奇B_3病毒(Cox B_3)心肌炎病理过程中脾NK细胞活性及血清、心肌组织匀浆中干扰素水平。在感染后的第3天,NK细胞活性及血清、心肌组织匀浆中干扰素滴度达高峰,并以较高水平维持到第7天。小鼠受染后3～7d心肌组织中病毒滴度亦达峰值。随着病毒从心肌中消失,NK细胞活性及干扰素滴度急剧降至对照组水平。实验结果提示:鼠Cox B_3感染早期,体内NK细胞活性和干扰素滴度升高的主要作用是溶解病毒感染细胞、限制病毒复制。

应用合成肽检测病毒性心肌炎患者血清中柯萨奇B组病毒抗体

目的研究应用合成肽代替完整病毒检测柯萨奇Ｂ组病毒（ＣＶＢ）抗体及其在病毒性心肌炎（ＶＭＣ）病因诊断中的作用。方法从ＣＶＢ衣壳蛋白ＶＰ１及ＶＰ３的保守区及ＣＶＢ３衣壳蛋白ＶＰ２的非保守区各选出一段多肽（分别称为ＶＰ１－１肽、ＶＰ３－１肽、ＶＰ２－１肽）进行化学合成，应用酶联免疫吸附法（ＥＬＩＳＡ）检测这几条多肽与型特异性ＣＶＢ１～６抗体的结合反应。以ＶＰ１－１肽、ＶＰ３－１肽作包被抗原，用间接ＥＬＩＳＡ检测患者血清中ＣＶＢＩｇＧ及ＶＣＢＩｇＭ抗体。结果ＶＰ１－１肽和ＶＰ３－１肽与型特异性ＣＶＢ１～６抗体均有良好的结合反应。８５例健康人、６７例急性ＶＭＣ、３９例慢性ＶＭＣ患者血清ＣＶＢＩｇＧ阳性率分别为２３．５％、５３．７％、３８．５％，ＣＶＢＩｇＭ阳性率分别为７．１％、４９．３％、２８．２％。ＣＶＢＩｇＭ抗体与中和抗体检测结果之间有良好的相关性（Ｐ＜０．００１），阳性率无明显差别（Ｐ＞０．０５）。结论应用合成肽抗原检测ＣＶＢＩｇＭ抗体有较好的敏感性和特异性。

贯众水提取液对感染柯萨奇B_3病毒的培养大鼠心肌细胞的影响

[目的 ]探讨贯众水提取液对感染柯萨奇B3 病毒的大鼠心肌细胞的抗病毒和保护作用 .[方法 ]实验取新生SD大鼠的乳鼠心室肌制备搏动心肌细胞 ,培养 48h后接种 10 3 TCID50 的柯萨奇B3 病毒做为实验性病毒性心肌炎模型 .[结果 ]贯众水提取液和炙甘草汤的同时加药组和 1h后加药组大鼠心肌细胞释放的乳酸脱氢酶和天冬氨酸转氨酶活性明显降低 ,感染 12h后加药组效果明显减弱 ,而 2 4h后加药组则无效 .[结论 ]贯众水提取液对感染柯萨奇B3 病毒的SD大鼠培养心肌细胞具有抗病毒和保护作用 。

111例成人急性柯萨奇病毒B心肌炎后室性早搏的长期随访

119例经血清学与临床诊断为急性柯萨奇病毒B心肌炎的病人,长期随访平均3.5年(4—98月),随访率93.3％。急性期后82％的患者有室性早搏长期间断发作。随访期间无一例发生昏厥、近乎昏厥、猝死或心电图证实的持久性室性心动过速发作,亦无扩张型心肌病的临床、X线或超声心动图表现。

麻疹样发疹患者柯萨奇B型病毒检测

目的:检测长春春季流行性皮肤斑丘疹患者外周血中柯萨奇B型病毒。方法:取56例春季流行性麻疹样发疹患者外周血标本,扩增目的DNA并进行特定限制性内切酶分析。结果:56份标本全部呈柯萨奇B6型病毒阳性。结论:柯萨奇B6型病毒可能是春季流行性麻疹样发疹的病原体。

大蒜素对柯萨奇病毒B_3感染的小鼠心肌炎免疫和心肌损伤的影响

目的探讨大蒜素对实验性小鼠病毒性心肌炎的自然杀伤(NK)细胞、T淋巴细胞活性以及心肌损伤的影响。方法采用200只Balb/c小鼠腹腔注射柯萨奇病毒B3(CVB3)建立实验性小鼠病毒性心肌炎模型。小鼠随机分为模型对照组、大蒜素小剂量组、大蒜素中剂量组、大蒜素高剂量组,每组50只。注射病毒后即刻给药,连续13d,于第1个小时、第1天、第3天、第7天、第14天处死存活鼠取材。观察NK细胞、T淋巴细胞活性及心肌病理变化,测定血清肌钙蛋白I(cTnI)含量。结果小鼠在感染CVB3后第1天,NK细胞活性增高,第3天后开始下降,第7天后达到最低;T淋巴细胞活性在第7天开始下降,随后维持在较低水平。心肌酶学在第7天增高有统计学意义。感染CVB3后的第3天后心肌内仅有少量炎症浸润,未有明显病变,第7天病变严重,心肌坏死和炎症浸润程度均较重,呈片状。小剂量的大蒜素可提高活性下降的NK细胞、T淋巴细胞的活性,减轻心肌细胞的病变,中剂量的大蒜素疗效最明显,大剂量的大蒜素未能随剂量的增加而提高疗效。结论大蒜素对实验性小鼠病毒性心肌炎具有治疗作用。

灭活CVB1在离体骨骼肌细胞中基因转移的研究

【目的】研究灭活后的柯萨奇病毒B1在原代培养骨骼肌细胞中对报道基因 pCDNA3 LacZ转移效率的影响。【方法】①用聚乙二醇沉淀和蔗糖低温超速离心法提纯并用 β 丙内酯灭活病毒 ;②分别用不同浓度的转铁蛋白聚赖氨酸 (TfpL) +LacZ、灭活CVB1+LacZ以及灭活CVB1+TfpL +LacZ在肌肉细胞进行基因转移 ,以报告基因产物 β 半乳糖苷酶染色阳性的细胞数作为基因转移效率指标。【结果】灭活CVB1+TfpL +LacZ在肌肉细胞中使基因转移效率由TfpL +LacZ组的不到1%提高到 15 %～ 2 0 %。【结论】灭活CVB1也能象腺病毒一样促进基因转移 ,CVB1的嗜肌肉特性为在肌肉组织中研究靶向明确的新载体提供了实验依据。

柯萨奇病毒B组感染与小儿过敏性紫癜发病的关系

目的 :为探讨柯萨奇病毒B组感染与小儿过敏性紫癜发病的关系。方法 :应用ELISA法检测 37例过敏性紫癜及 4 0例上呼吸道感染患儿血清中柯萨奇病毒B组特异性抗体 (CBV -IgM)。结果 :过敏性紫癜组血清CBV -IgM阳性 17例 (4 5 .95 % ) ,对照组阳性 4例 (10 .0 0 % ) ,两组比较差异有非常显著性 ,X2 =12 .5 2 ,P <0 .0 1。 37例过敏性紫癜患儿中血清Adv及RSV阳性各 2例 ,CMV抗体均阴性。结论 :柯萨奇病毒B组感染与小儿过敏性紫癜发病密切相关 ,可能是过敏性紫癜的病因之一。

心肌尔康对柯萨奇B_（3m）株的抑制作用

用细胞病变抑制试验研究心肌尔康抗柯萨奇Ｂ３ｍ的体外作用。发现心肌尔康作１０倍系列稀释试验时，对１和１０ＴＣＩＤ５０病毒的５０％抑制（ＩＣ５０）剂量分别为１．３４×１０－２ｇ·Ｌ－１和１．５４ｇ·Ｌ－１，对１００和１０００ＴＣＩＤ５０病毒不能测出ＩＣ５０；该药在２倍系列稀释时，对１０和１００ＴＣＩＤ５０病毒的ＩＣ５０分别为１．８１ｇ·Ｌ－１和２．５８·Ｌ－１，对１０００ＴＣＩＤ５０病毒仍无抑制，试验表明，心肌尔康在体外对柯萨奇Ｂ３ｍ病毒有一定的抑制作用。

柯萨基B组病毒所致病毒性心肌炎间接ELISA检测方法的建立

利用提纯的Cox B1～6型病毒抗原分别包板、建立了检测临床诊断为病毒性心肌炎患儿血清中特异性IgM的间接ELISA方法。32例患儿血清中有23例检为阳性(71.9％)。利用提纯的兔抗Cox B1～6免疫血清分别包板来捕捉病毒抗原的改进间接ELISA方法检测32例患儿血清,有28例检为阳性(87.5％)。用上述两种方法对30份正常人血清进行检测,均有1份检出阳性(3.3％)。与白求恩医科大学提供的检测药盒比较,本实验方法具有较高特异性,且敏感性不受类风湿因子(RF)的影响。