人源细粒棘球蚴染色体G-和C-带初步分析

人源细粒棘球蚴染色体数目初步测定为 14～ 18条[1 ] 。本实验进一步对染色体Ｇ 带和Ｃ 带作了研究。 军队医药卫生青年基金资助课题 (Ｎｏ .96Ｑ0 2 3) 现单位 :中山医科大学基础医学博士后流动站 ,广州 5 10 0 891　材料与方法1 1　培养材料的制备　临床手术确诊的肝包虫 (细粒棘球蚴 )病患者 ,女性 ,包囊直径为 10ｃｍ ,按参考文献 [2 ]制备生发层细胞和原头节培养材料用于培养 ,并制备染色体标本。1 2　Ｇ 带的制备方法　将干燥的未染色的染色体片置于磷酸缓冲液 (ＰＢ)中湿润 ,6 0℃温育 10ｍｉｎ ;空气中自然干燥 ,每张片滴加 1ｍｌ…     

4种免疫原对棘球绦虫继发性感染的预防效果

由于猪带绦虫和细粒棘球绦虫间存在大量的共同抗原〔1～ 3〕,为寻找可能的免疫原制备细粒棘球绦虫疫苗 ,我们将 4种猪带绦虫和细粒棘球绦虫抗原进行了初步免疫预防的动物实验 ,现将结果报告如下1　材料与方法1 1　免疫原制备　Ｅ ｇＣＦ免疫原 :细粒棘球蚴囊液采绵羊肝脏 ,离心     

酯酶同工酶鉴定细粒棘球蚴细胞系(13G-5)种属来源

目的　鉴定细粒棘球蚴细胞系细胞的种属来源。方法　应用SDS凝胶电泳测定细粒棘球蚴细胞系酯酶同工酶 (EST) ,并和E granulosus原头节、囊液及包虫病人血清作比较。结果　细粒棘球蚴细胞系、E granulosus原头节、囊液含有相同的酯酶同工酶谱 ,在同一位点出现相同的一条酶带。结论　实验结果表明从包虫病人培育的 13G - 5细胞确系来源于E granulosus细胞。     

RAPD技术在宁夏人源细粒棘球蚴基因分型中的应用

目的建立RAPD分子分型技术并将其应用于人源细粒棘球蚴基因分型研究。方法用单因素筛选方法建立PCR-RAPD分析体系,并用建立的RAPD分析方法对分离自宁夏不同地区13例包虫病病人的细粒棘球蚴进行RAPD指纹图谱分析。结果采用20条随机引物对细粒棘球蚴DNA进行扩增,其中11条引物呈现良好的多态性和稳定性,共得到232条片段。根据RAPD指纹图谱将13株人源细粒棘球蚴分为Ⅰ型(53.85%)、Ⅱ型(23.08%)、Ⅲ型(7.69%)和Ⅳ型(15.38%)4个种内基因类群。结论宁夏地区人源细粒棘球蚴之间存在不同的种内遗传差异。     

感染细粒棘球蚴对宿主(羊)肝药酶的影响及肝组织超微结构的变化

利用生化、组化方法测定肝微粒体中细胞色素P-450、bs含量及NADPH细胞色素P-450还原酶，研究细粒棘球蚴感染对宿主（羊）肝药酶的影响及其变化规律，并对肝组织进行光镜、电镜观察。结果表明细粒棘球蚴感染可引起肝组织损伤并降低肝药酶活性，对肝药酶的抑制作用与距棘球蚴的远近呈梯度变化即距棘球蚴越近酶活性降低越明显，肝组织损伤并降低肝药酶活性，对肝药酶的抑制作用与距棘球蚴的远近呈梯度变化唧 距棘球蚴越近酶活性降低越明显。肝组织受损程度与肝药酶的活性降低程度一致，电镜下可见细凿棘球蚴周边肝细胞粗面内质网明显脱颗粒。     

高强度聚焦超声辐照后细粒棘球蚴囊壁的病理变化

目的 观察细粒棘球蚴囊壁在高强度聚焦超声（HIFU）辐照后的病理改变。方法 采集感染细粒棘球蚴的新鲜羊肝，选取囊壁较薄、触摸弹性较好的细粒棘球蚴30个。采用随机抽样方法等分为3组，每组10个包囊。对照组，用普通诊断超声照射2 min。处理组1和处理组2分别用150 W和250 W声功率对细粒棘球蚴包囊进行沿囊壁多层面的环形扫描，层面间距为5 mm，扫描速度为3 mm/s，照射时间2～10 min（根据包囊大小）。取出照射后先肉眼观察细粒棘球蚴包囊大体改变，后取囊壁组织分别制作病理切片和透射电镜切片，观察其病理改变。 结果 HIFU（250 W）辐照后，细粒棘球蚴包囊剪开处内囊壁立即发生卷曲，剥离出的内囊颜色变白、变硬、透光度降低。病理切片显示，HIFU辐照后细粒棘球蚴的内囊壁上角皮层与生发层大部分发生分离。电镜观察结果显示，HIFU辐照后细粒棘球蚴的角皮层纤维纹理明显改变，生发层细胞发生裂解性破坏。 结论 HIFU沿细粒棘球蚴囊壁的多层面的环形照射可明显损害细粒棘球蚴囊壁。     

细粒棘球蚴生发细胞体外培养的实验观察

目的 :为培育人源细粒棘球蚴细胞系 (株 ) ,以供用于免疫预防研究。方法 :用外科方法从临床确诊的细粒棘球蚴患者的肝脏中取出包囊 ,以生发层和原头节为培养材料 ,采用RPMI1640、199和改良 DMEM培养液 ,用鼠尾胶原蛋白包被和不包被的培养瓶 ,对其进行初培养 ,并进行对比观察。结果 :以胶原蛋白作为支撑材料 ,应用改良 DMEM培养液培养的人源细粒棘球蚴生发层和原头节效果优于其他 ,已成功地培育了人源细粒棘球蚴细胞系 ,并传至第 2 0代。原代培养时间需 2 8d- 4 5d,3代以内细胞为多形态性。结论 :建立了适合人源细粒棘球蚴细胞体外培养的方法。     

细粒棘球蚴生发细胞体外培养的实验观察

目的 :为培育人源细粒棘球蚴细胞系 (株 ) ,以供用于免疫预防研究。方法 :用外科方法从临床确诊的细粒棘球蚴患者的肝脏中取出包囊 ,以生发层和原头节为培养材料 ,采用RPMI1640、199和改良 DMEM培养液 ,用鼠尾胶原蛋白包被和不包被的培养瓶 ,对其进行初培养 ,并进行对比观察。结果 :以胶原蛋白作为支撑材料 ,应用改良 DMEM培养液培养的人源细粒棘球蚴生发层和原头节效果优于其他 ,已成功地培育了人源细粒棘球蚴细胞系 ,并传至第 2 0代。原代培养时间需 2 8d- 4 5d,3代以内细胞为多形态性。结论 :建立了适合人源细粒棘球蚴细胞体外培养的方法。     

绵羊源细粒棘球蚴原头节感染子午砂土鼠和小鼠的比较观察

子午砂土鼠，ＮＩＨ小鼠和昆明株小鼠在实验感染绵羊源细粒棘球蚴原头节后，在不同时期剖检，观察了细粒棘球蚴在其体内的发育状况，三种鼠平均湿囊重及其占体重百分比，子午砂土鼠均高于其他两种小鼠。子午砂土鼠在感激后２７４天时，部分原头节已发育成熟，在３５７天时，全部成熟，结果表明，子午砂土鼠对细粒棘球蚴的敏感性高于其他小鼠，表现为包囊发育好，生长快，是细粒棘球蚴的一种比较理想的实验动物。     

细粒棘球绦虫水通道蛋白9的多肽特征及免疫定位研究

目的 研制细粒棘球绦虫水通道蛋白9 (EgAQP9)的特异性多肽抗体,并用于检测其在棘球蚴囊壁、生发细胞及原头蚴中的组织分布情况。 方法 根据EgAQP9特异氨基酸序列设计合成B细胞抗原多肽,再用合成的多肽偶联KLH作为免疫原注射免疫新西兰兔以制备多克隆抗体。酶联免疫吸附试验(ELISA)方法检测多肽抗体滴度,蛋白免疫印迹法(Western blot)测定多肽抗体免疫活性,免疫荧光实验分析EgAQP9的亚细胞定位,免疫组化实验分析EgAQP9组织定位。 结果 免疫新西兰兔成功制备出多肽抗体;间接ELISA检测其多肽抗体效价高达1∶256 000;Western blot结果显示多肽抗体能够特异识别细粒棘球绦虫中36 kDa处的条带,与EgAQP9预测的相对分子量大小相符;免疫荧光实验结果显示EgAQP9分布于棘球蚴生发细胞的胞质、胞膜中;免疫组化实验显示该蛋白主要分布在原头蚴表面及棘球蚴囊泡生发层。结论 本研究以制备的EgAQP9兔源多克隆抗体定位了AQP9在棘球蚴生发细胞、原头蚴及棘球蚴囊壁中的分布状况。     

Short report: Inadequacy of yaks as hosts for the sheep dog strain of Echinococcus granulosus or for E. Multilocularis

Hydatid disease (echinococcosis) has a two-host cycle involving the domestic dog and grazing animals. Humans are also infected by the dog. Both unilocular (Echinococcus granulosus in yaks, sheep, and goats) and multilocular (alveolar) (E. multilocularis in hares and rodents) hydatids are common in western Sichuan in the People's Republic of China. Humans and dogs are equally infected with both species. Many yaks (Bos grunniens) were found with multilocular cysts that visually were deemed to be E. multilocularis. However, a histologic and molecular study showed that they were actually E. granulosus. No infective cysts were found in 125 necropsied yaks. We conclude that the yak is an inadequate and dead-end host for the sheep dog (G1) strain of Echinococcus granulosus and also for E. multilocularis.     

放射线治疗包虫病的实验研究

目的通过观察不同剂量6MV X线照射对大鼠继发性棘球蚴活性的影响,探讨放射线治疗棘球蚴病的可行性及其机制。方法对从大鼠体内分离出的棘球蚴囊分别以10、20、30和40Gy的6MV X线进行照射后,观察棘球蚴囊的超微结构,测定囊液中总蛋白和钙离子含量,并通过草酸盐一焦锑酸盐细胞化学法(PPA)探讨放射线治疗包虫病的机理。结果与空白对照组相比,各实验组棘球蚴囊超微结构均可见不同程度的改变和破坏;囊液总蛋白含量明显降低而钙离子含量明显增高,且以高剂量组更为显著。PPA法显示实验组内质网与线粒体扩张且膜上可见黑色光滑的钙盐沉积。结论6MV X线对棘球蚴囊有一定破坏作用,且在一定的范围内存在量效关系。     

''Arc 5'' antibodies in sera of sheep infected with Echinococcus granulosus, Taenia hydatigena and Taenia ovis

The 'arc 5' precipitin band, formed when test human serum is reacted against immunoelectophoresed hydatid cyst fluid antigen, has provided a positive diagnosis of Echinococcus granulosus infection. These antibodies to 'arc 5' antigen have now been found in the sera of sheep. They appear 2 weeks after infection with Taenia ovis, after 3 weeks with T. hydatigena and after 16 weeks with E. granulosus. An antigen similar to the 'arc 5' antigen of E. granulosus cyst fluid was also demonstrated in cyst fluid from T. hydatigena, but it would not be positively identified in T. ovis cyst fluid. The presence of 'arc 5' in immunoelectrophoresis tests is not suitable for specific immunodiagnosis of E. granulosus infections in sheep in New Zealand. 'Arc 5' antibodies were only associated with living E. granulosus cysts and were not present if cysts were dead. The location of the cysts in either liver or lungs and the onset of brood capsule production did not influence the presence of 'arc 5' antibodies.     

小鼠细粒棘球蚴体外促血管生成作用研究

目的分析细粒棘球蚴体外对人脐静脉内皮细胞血管生成能力的影响及其促血管生成因子转录水平,初步探讨细粒棘球蚴的促血管生成作用。方法将继发感染小鼠体内的细粒棘球蚴体外培养后的上清作用于体外培养的人脐静脉内皮细胞,观察其成管现象并用NIH Image J软件的Angiogenesis模块进行分析。同时,在细粒棘球蚴基因组中寻找同宿主(小鼠)的基质金属蛋白酶9(Matrix metalloproteinase-9,MMP-9)和高迁移率族蛋白B1(High mobility group box B1,HMGB1)的同源蛋白,并对其转录水平进行分析。结果鼠源细粒棘球蚴囊培养上清能显著促进人脐静脉内皮细胞的成管(F=73.03,P0.001),在细粒棘球蚴囊壁和原头节中存在小鼠MMP-9和HMGB1同源蛋白的表达,且MMP-9在原头节中的转录水平高于棘球蚴囊(t=-11.65,P0.001),而HMGB1在棘球蚴囊中的转录水平高于原头节(t=6.43,P=0.003)。结论细粒棘球蚴囊培养上清可能含有促进血管生成的虫源分子,但其确切机制还有待进一步研究。     

The epidemiology of Echinococcus granulosus in Great Britain. V. The status of subspecies of Echinococcus granulosus in Great Britain

Twenty-five years ago Williams and Sweatman suggested that in Great Britain there are two subspecies of Echinococcus granulosus--E. granulosus granulosus and E. granulosus equinus. Echinococcus granulosus granulosus does not mature either in foxes or in horses: E. granulosus equinus will mature in either. The prepatent period of E. granulosus granulosus in the definitive host is about 42 days while that of E. granulosus equinus is about 70 days. Each subspecies has a characteristic morphology. More recently, in the course of seven experiments, dogs, red foxes (Vulpes vulpes crucigera), arctic foxes (Alopex lagopus), badgers (Meles meles), domestic ferrets and domestic cats have been infected with protoscoleces derived from hydatid cysts of human, equine and ovine hosts from different regions of England and Wales. Transmissions to horses and sheep were always succeeded by the development of viable hydatid cysts; transmissions to dogs and foxes, by the development of gravid adults. The prepatent period in both dogs and foxes was invariably about 70 days (or longer) and the morphology of all the adult parasites in these definitive hosts was characteristic of E. granulosus equinus. All the evidence derived from these experiments supports the view that there is, in Great Britain, but one subspecies of E. granulosus. That one subspecies is E. granulosus equinus. It infects dogs, red foxes, arctic foxes (experimentally), cats (experimentally), humans, sheep, goats, horses, donkeys, pigs, cattle, roe deer and reindeer (in Scotland). Attempts to transmit E. granulosus to badgers and domestic ferrets were unsuccessful. Of 123 cats infected with protoscoleces of horse origin, one gravid adult parasite was recovered from one animal.     

An improvement in a new method for the surgical treatment of echinococcosis

Parasitic cysts were intraoperatively treated with glycerol in 179 patients with echinococcosis at various sites and 3% hydrogen peroxide in 165 patients with hepatic echinococcosis. Experiments on 60 cotton rats experimentally infected with Echinococcus alveolaris and 156 albino mice with E. granulosus provided evidence for the surgical use of 80-100% glycerol or 3% hydrogen peroxide to treat the cysts. Intraoperative glycerol treatment of cysts in patients with complicated hepatic echinococcosis fails to normalize amino acid metabolism, as well as immunity even a year postoperatively. The glycerol method has no contraindications for pulmonary and hepatic echinococcosis. Hydrogen peroxide has a damaging effect on all germinal elements of both E. alveolaris and E. granulosus. The authors recommend that 3% hydrogen peroxide should be intraoperatively used for treatment of cysts. After surgery, there is a tendency for amino acid metabolism to become normal in such patients.     

Clinical value of Echinococcus granulosus--specific immunoglobulin G antibody avidity in liver cystic echinococcosis

Role of E. granulosus-specific IgG antibody avidity in evaluation of a natural history of the parasite cysts in the liver, duration of infection and immunogenic cysts activity was studied. Thirty-one patients with liver cysts suspected of echinococcosis were examined; E. granulosus infection was finally confirmed by a fine needle aspiration biopsy or surgery in 29 cases (93.5%) and two non-parasitic simple cysts in the liver were diagnosed. E. granulosus-specific IgG antibody was found in 21 patients (67.7%). High IgG avidity values were observed in 14 patients with liver cysts in a degeneration stage confirmed by imaging techniques. Low IgG avidity values were mostly detected in children with hypoechogenic echinococcal cysts being in a recent stage of infection. Determination of anti-E. granulosus immunoglobulin G antibody avidity is a very usefull diagnostic tool for a determination of parasitic cysts activity and their developmental age.     

吡喹酮在体内,体外抗细粒棘球蚴囊的作用（摘要）

吡喹酮在体内、体外抗细粒棘球蚴囊的作用（摘要）肖树华，杨元清，尤纪青，沈炳贵，柴君杰（中国预防医学科学院寄生虫病研究所，世界卫生组织疟疾、血吸虫病和丝虫病合作中心，上海２０００２５，中国；新疆地方病防治研究所，乌鲁木齐８３０００２，中国）关键词　细粒...     

Human cystic echinococcosis: epidemiologic, zoonotic, clinical, diagnostic and therapeutic aspects

This review represents an updated scenario on the transmission cycle, epidemiology, clinical features and pathogenicity, diagnosis and treatment, and prevention and control measures of a cestode parasite Echincoccus granulosus (E. granulosus) infection causing cystic echinococcosis (CE) in humans. Human CE is a serious life-threatening neglected zoonotic disease that occurs in both developing and developed countries, and is recognized as a major public health problem. The life cycle of E. granulosus involves a definitive host (dogs and other canids) for the adult E. granulosus that resides in the intestine, and an intermediate host (sheep and other herbivores) for the tissue-invading metacestode (larval) stage. Humans are only incidentally infected; since the completion of the life cycle of E. granulosus depends on carnivores feeding on herbivores bearing hydatid cysts with viable protoscoleces, humans represent usually the dead end for the parasite. On ingestion of E. granulosus eggs, hydatid cysts are formed mostly in liver and lungs, and occasionally in other organs of human body, which are considered as uncommon sites of localization of hydatid cysts. The diagnosis of extrahepatic echinococcal disease is more accurate today because of the availability of new imaging techniques, and the current treatments include surgery and percutaneous drainage, and chemotherapy (albendazole and mebendazole). But, the wild animals that involve in sylvatic cycle may overlap and interact with the domestic sheep-dog cycle, and thus complicating the control efforts. The updated facts and phenomena regarding human and animal CE presented herein are due to the web search of SCI and non-SCI journals.     

细粒棘球蚴和猪囊尾蚴的组织特征及其对化学疗法的影响

对细粒棘球蚴及猪囊尾蚴的组织结构进行了观察和比较。结果发现：两者的主要区别是前者具有宽厚的角质层，且含有少量血管的纤维外囊又远离囊腔，而后者则无角质层结构，且含有丰富血管的纤维外囊又紧密毗邻囊腔。上述区别为抗绦虫药对两种后绦期幼虫疗效差异的分析，提供了组织病理学的依据。