鸭乙型肝炎病毒对鸭心肌细胞超微结构的影响

将鸭乙型肝炎病毒(DHBV)感染雏鸭制作的鸭乙型肝炎模型已被广泛用于研制抗乙肝病毒新药,为探索DHBV感染对雏鸭心脏有何影响,我们观察了被DHBV感染的雏鸭其心肌的超微结构,现报道如下。 1 材料与方法 1.1 动物及分组 20只1日龄武汉麻鸭(购自华中农业大学种禽厂),随机分为对照组和感染组,每组10只。 1.2 造模方法及标本制作 雏鸭喂养1周后,分成2组:感染组每只鸭自胫静脉注射DHBV阳性血清     

鸭乙肝病毒经不同途径感染雏鸭的造模研究

取1～3日龄健康雏麻鸭67只予以腿部肌注 DHBV,1周后感染率达88.0%,并持续3个月以上.电镜下可见血清阳性鸭的肝细胞胞浆中散在有 DHBV 的完整颗粒和内质网中密集的空泡颗粒。另将14日龄鸭28只分别予 DHBV 静脉注射或肌内注射,1周后血清阳转率各为35.7%和42.9%,持续1周又全部转阴;肝组织病理学观察,阳性鸭炎症改变明显.此外,对1～15日龄雏鸭94只,经 DHBV 灌胃后.连续采血测定均阴性。本研究表明,DHBV腿部肌注是最简便可靠的实验感染途径.     

中药复方乙肝解毒丸抗DHBV的实验研究

以人工感染鸭乙型肝炎病毒(DHBV)的雏鸭为实验模型,以斑点杂交试验检测 DHBV DNA 为实验指标,用乙肝解毒丸1号(由大黄、虎杖、羊蹄根、山豆根等组成)、Ⅱ号(由土茯苓、红藤、苦参、猪苓等组成)连续给药二个月进行抗 DHBV 实验治疗。结果:Ⅰ号方治疗组转阴率为29.54％(13/44);Ⅱ号方治疗组转阴率为34.04％(16/47);对照组转阴率为7.89％(3/38),P 分别<0.05和 P<0.01.提示两中药复方均具有抗 DHBV 治疗作用。     

小分子肽抗乙型肝炎病毒的实验研究

目的:应用2.2.15细胞株和鸭乙型肝炎病毒(DHBV)感染模型,评价小分子肽CMS 017抗乙肝病毒作用.方法:①CMS 017以倍比稀释浓度加入2.2.15细胞培养中,作用8天后吸取培养上清,PCR法检测HBVDNA.②建立DHBV感染模型,CMS017以60、200、600μg/kg·d-13个剂量腹腔注射给药28天,检测用药前、后血清DHBV DNA、DHBsAg变化.结果:①CMS 017体外抑制DHBV呈量效关系,IC50为2.3μg/ml.②鸭体内实验中,CMS 017中、高剂量组用药7天、14天开始出现血清DHBV DNA降低(P＜0.01,P＜0.05),持续至停药后7天.CMS 017中剂量组用药后血清DHBsAg降低的时效关系与DHBV DNA完全一致.结论:CMS017具有抗乙肝病毒作用,其体内作用的量效关系待确定.     

鸭乙型肝炎肝纤维化模型的研制(摘要)

目的:采用鸭乙型肝炎病毒(DHBV)阳性血清反复攻击雏鸭建立鸭乙型肝炎肝纤维化动物模型。方法:用DHBV阳性血清0.1ml/只,从胫静脉注射1日龄樱桃谷鸭,每周1次,从第10周剂量加大为     

广西麻鸭乙型肝炎感染病毒动物模型的实验研究

目的探讨广西麻鸭作为乙型肝炎病毒感染动物模型的可能性。方法应用广西1 d龄雏麻鸭经腹腔感染鸭乙型肝炎病毒(DHBV),13 d后采用实时荧光定量PCR法检测麻鸭血清DHBV含量,筛选出DHBV强阳性鸭。结果共检测148份麻鸭血清标本,其中DHBV阳性标本136份,阳性率为91.9%。结论广西麻鸭可作为乙型肝炎病毒感染动物模型。实时荧光定量PCR法检测DHBV敏感性较高,重复性好,可用于DHBV检测。     

海珠益肝胶囊抗鸭乙型肝炎病毒的实验研究

目的:研究海珠益肝胶囊对鸭乙型肝炎的抗病毒作用.方法:使1日龄北京麻鸭感染鸭乙型肝炎病毒(DHBV),7天后用Dot-EIA法筛选出DHBsAg强阳性鸭.随机分成5组:海珠益肝胶囊大、中、小3个剂量组、以生理盐水灌胃的模型组和以无环鸟苷(ACV)灌胃的对照组.每组6只,各组雏鸭均灌胃10天.用药前、用药5天、10天及停药后3天分别采血,采用斑点杂交法检测血清DHBV DNA.结果:用海珠益肝胶囊5天、10天时鸭血清DHBV DNA其OD值均明显低于给药前,差异有显著性意义(P＜0.05).与对照组相比,差异无显著性意义(P＞0.05).结论:海珠益肝胶囊有一定的抑制DHBV DNA复制的作用.     

护肝煎剂抑制鸭肝炎病毒的实验研究

鸭乙型肝炎病毒(DHBV)是1980年才发现的嗜肝DNA病毒,与人乙型肝炎病毒(HBV)的生物学特征及其与宿主肝脏疾病的关系十分相似。国内外近10年来的研究表明,DHBV感染的鸭子可以用作研究人乙型肝炎和肝癌较为理想的动物模型。本研究是用含有DHBV的麻鸭血清注射一日龄鸭足静脉造成人工感染,用具有抗HBV、活血化瘀、调节免疫功能等作用中药制成护肝煎剂进行实验治疗,以检测鸭血清中DHBeAg的P/n值作为疗效指标,研究结果报告如下。     

补锌去铁对中药保肝康抗鸭乙肝病毒疗效的影响(摘要)

目的:观察补锌去铁对中药保肝康抗鸭乙型肝炎病毒(DHBV)疗效的影响。方法:采用垂直传播的DHBV DNA阳性的6月龄麻鸭作鸭乙型肝炎病毒模型。以补锌去铁加中药保肝康进行治疗,观察其对DHBV、鸭肝功能及血清锌、铁、肝组织铁的影响。并设保肝康作为中药对照,抗乙肝转移因子作     

肝苏颗粒浸膏粉抗鸭乙型肝炎病毒的实验研究

目的：观察肝苏颗粒浸膏粉（以下简称肝苏）体内抗鸭乙型肝炎病毒（DHBV）的作用,为临床治疗乙肝病毒感染提供实验依据,方法：采用重庆鸭乙型肝炎动物模型,用肝苏口服治疗4W,停药观察1W,检测用药前后血清的DHBVDNA,DHBsAg,转氨酶（ALT,AST）及肝组织病理变化,结果：肝苏各剂量组用药后血清DHBV DNA滴度显著降低或极显著降低（P＜0．05或P＜0．01）,停药1W后中,小剂量组血清DHBV DNA有明显回升现象,而大剂量组DHBV DNA回升现象不明显,其抗病毒效果与剂量大小有一定的关系,用药前后血清DHBsAg O,D值（490nm)的变化与DNA滴度改变相似,此外,肝脏病理学检查及治疗4W,停药1W后血清ALT,AST检查未发现该药对鸭肝组织有明显的毒性损害,结论：连续用肝苏1个月在鸭体内有抗鸭乙型肝炎病毒的作用。     

肝康栓抗鸭乙型肝炎病毒和改善肝脏病理的作用

[目的]用鸭乙型肝炎（乙肝）模型研究肝康栓抗鸭乙型肝炎病毒（DHBV）和改善肝脏病理的作用。[方法]用DHBV阳性血清感染1d龄的樱桃谷雏鸭，制备鸭乙肝模型。将其中36只感染阳性鸭随机分成3组：肝康栓治疗组，0．85％氯化钠模型组和阿昔洛韦（ACV）对照组，每组12只，均连续给药4周。分别于给药前，给药14、28d和停药7d时取血清，用实时定量PCR法检测鸭血清中DHIWDNA拷贝数。于停药7d时，处死各组雏鸭，取肝脏病理切片苏木精一伊红染色后，观察肝脏炎症情况和肝细胞变性程度。[结果]肝康栓治疗组给药14、28d及停药7d时鸭血清中DHBVDNA拷贝数的对数值均较给药前降低（均P〈0.01）。肝康栓治疗组雏鸭肝脏的肝细胞肿胀率、空泡变率、嗜酸性变率均较同期模型组降低，差异有统计学意义（P〈0.05，〈20．01）。[结论]肝康栓能有效地抑制鸭体内DHBVDNA的复制，并具有改善肝脏病理的作用。     

919糖浆抗鸭乙型肝炎病毒的实验研究

目的:研究919糖浆时鸭乙型肝炎的体内抗病毒作用。方法:以感染鸭乙型肝炎病毒(DHBV)的武汉麻鸭为实验动物模型。使1d龄武汉麻鸭感染DHBV,7d后用Dot-EIA法筛选出DHBsAg强阳性鸭。随机分组后给予919糖浆治疗14d,于感染后第7d(为用药前)、用药7d、14d及停药后3d分别采血,采用斑点杂交方法检测血清DHBVDNA。结果。用药919糖浆7d、14d鸭血清DHBV DNA OD值均明显低于给药前,有显著性差异(P<0.05,P<0.01)。停药3d后没有反跳现象。结论:919糖浆有一定的抑制DHBV DNA复制作用。     

肝康栓抗鸭乙型肝炎病毒的实验研究

目的：用鸭乙肝模型研究肝康栓对鸭乙型肝炎病毒（DHBV）的体内抗病毒作用。方法：用DHBV阳性血清感染1日龄的樱桃谷鸭，制备鸭乙型肝炎模型。随机分成5组：肝康栓大、中、小3个剂量治疗组，生理盐水模型组和阿昔洛韦（ACV）对照组，每组12只，均连续给药4周。分别于给药前，给药14天、28天，停药7天时取血清，用real-timePCR法检测鸭血清中DHBV DNA拷贝数的变化情况。结果：肝康栓大剂量组给药14天、28天及停药7天，中剂量组给药14天、28天，小剂量组给药28天，鸭血清中DHBV DNA拷贝数的对数值均较给药前降低（〉2个对数级），差异有统计学意义（P〈0．01）。结论：肝康栓具有有效抑制鸭体内DHBV DNA复制的作用。     

扇贝多糖抗鸭乙型肝炎病毒作用

摘要： 目的 研究扇贝多糖抗鸭乙型肝炎病毒（DHBV）的作用。方法 选1日龄北京鸭,人工感染鸭乙型肝炎病毒（DHBV）,感染后第7天,阳性鸭随机分为模型组、拉米夫定阳性对照组（50 mg.kg-1）和扇贝多糖（75、150、300 mg.kg-1）3个剂量组,均连续灌胃给药10d,每日2次。分别于给药前、给药5d、10d及停药后3d取血清,应用斑点杂交法检测血清中 DHBV-DNA水平。结果 扇贝多糖150、300mg.kg-1剂量组于用药后5d、10d,与给药前（T0）比较,血清DHBV-DNA水平明显降低（P<0.05、P<0.01）。结论 扇贝多糖体内有抑制DHBV-DNA的作用。     

Studies on duck hepatitis B virus: Identification and epidemiological survey in Taiwan

The presence of duck hepatitis B virus (DHBV) in domestic ducks in Taiwan was confirmed by DNA polymerase assay, Southern blot analysis and electron microscopy. To investigate the epidemiology of this virus, a total of 1274 serum samples were collected from 30 duck farms from different areas of Taiwan and studied by spot hybridization and/or DNA polymerase assay. The positive rates varied among different strains of ducks: 16% in 243 Pekin ducks, 12% in 392 Chinese common domestic ducks, 4% in 196 Muscovy, 25% in 292 Taiwan Kaiyas and 13% in 151 mule ducks. The positive rate was much higher in the younger ducks; it was highest (30.7%) in ducklings under 1 month of age, followed by ducks aged 1–12 months (11.8%), and lowest in those ducks older than 1 year (7.7%). It was concluded that the prevalence of DHBV infection in domestic ducks in Taiwan is generally high, and that the infected ducks may serve as an animal model for human hepatitis B virus infection which is also prevalent in Taiwan.     

Liver disease associated with duck hepatitis B virus infection of domestic ducks.

The liver disease associated with duck hepatitis B viremia was investigated in naturally infected ducks from Chi-tung county in China and in both naturally and experimentally infected ducks from the United States. Liver and serum specimens of adult Chinese ducks were examined for duck hepatitis B virus (DHBV) DNA by dot and gel blot hybridization. DHBV was found in serum and (in episomal form only) in livers of 6 of 11 birds exhibiting various degrees of chronic hepatitis. In 1 bird with hepatocellular carcinoma, DHBV DNA was detected at the limit of assay sensitivity and in another not at all, contrasting with findings in humans and woodchucks. In work with California Pekin and Khaki Campbell ducks, known amounts of DHBV were injected into the egg 10 days before, or into ducklings 1 day after, hatching and the livers were examined 6 weeks later. The majority of the injected ducklings had viremia detectable by hybridization 1 or 2 weeks after injection. The presence but not the amount of viremia correlated with incidence and degree of hepatitis, determined under code. The most severe instances of hepatitis, all in Pekin ducks, resembled the hepatitis in adult Chinese ducks of Chi-tung county. Severe and moderate hepatitis were found only in indoor-caged injected animals with viremia and in some uninjected birds without viremia that had been kept in outdoor flocks. The latter hepatitis, as some hepatitis in adult Chinese ducks, may not be related to DHBV. Mild and insignificant hepatitis were also found in injected and noninjected ducklings, some of which had the vertically transmitted spontaneous viremia previously described. The good correlation of experimentally induced viremia with incidence and severity of hepatitis in the Pekin duckling provides a simple, rapid, and relatively inexpensive model to study the relation of lesions to hepatitis B family infection in nonprimates.     

Duck hepatitis B virus infection of non-hepatocytes

One hundred and seventeen ducklings, 42 inoculated with duck hepatitis B virus (DHBV) 2 days after hatching and 55 connatally infected, were studied over a 6-month period in parallel with 20 ducklings without DHBV infection. Using immunohistochemical, in situ and blot hybridization analyses, the natural course of hepatic and extrahepatic infection was examined. DHBV infection started in the liver 2-4 days post-inoculation. There, DHBV was found not only in hepatocytes, but also in bile duct epithelial cells. Further, DHBV infection occurred in exocrine and endocrine pancreas (beginning 6-10 days and 20 days post-inoculation, respectively) and in germinal centers of the spleen (beginning 8 weeks post-inoculation). Occasionally viral DNA was also found in kidney glomeruli. Using strand-specific RNA probes, viral DNA in pancreas and spleen was clearly demonstrated to be replicating intermediates. Hepatic and extrahepatic infection with DHBV was not associated with histologic inflammation or pathologic changes in these tissues or the liver. These data indicate that DHBV can infect cells other than hepatocytes. The biological significance of non-hepatocyte infection for the life-cycle of the virus and its potential significance for viral persistence remain to be determined.     

Demonstration of duck hepatitis B virus in bile duct epithelial cells: Implications for pathogenesis and persistent infection

Hepatitis B virus (HBV) has been demonstrated in bile duct epithelial cells (BDEC) during chronic infection. The persistence of virus in BDEC may play an important role in disease pathogenesis, and may be at least partly responsible for the relapse phenomenon observed in antiviral treatments using nucleoside analogues. The aims of this study were to examine the morphological changes within the liver in the duck hepatitis B model following bile duct ligation (BDL), and to assess the effect of biliary hyperplasia upon viral DNA and proteins. Seven-day-old ducklings, congenitally infected with the duck hepatitis B virus (DHBV), were subject to BDL. The pathological and virological changes were then followed at 5, 10, 15, and 20 days after ligation. All results were compared with age-matched unligated control birds congenitally infected with DHBV. To assess the early morphological changes, additional animals were sacrificed at 1, 2, 3, and 4 days post-BDL. The proportion of DHBV-infected BDEC, was examined by immunohistochemistry and in situ hybridization. BDL induced rapid biliary hyperplasia, with a doubling time for BDEC of 1.3 days. The proliferated BDEC displayed immunohistochemical features identical to resting BDEC. More than 50% of BDEC in unligated controls, and more than 46% of proliferated BDEC in ligated animals were positive for DHBV DNA and structural proteins. The intensity of immunohistochemical staining and in situ hybridization signal in the BDEC was consistently greater than that of the hepatocytes, both before and after BDL. BDL induces biliary hyperplasia in the duck model, and BDEC division does not reduce the viral burden in infected cells.(Hepatology 1997 Feb;25(2):463-9)