沙鼠肝泡球蚴组织中骨桥蛋白与IL-10的表达及相关性研究

目的 检测沙鼠肝泡状棘球蚴组织中骨桥蛋白（OPN）及白细胞介素-10（IL-10）的表达,并进一步探讨两者的相关性。方法 20%泡状棘球蚴组织混悬液开腹直视下肝脏穿刺注射感染长爪沙鼠60只,每只0.1mL,建立肝泡状棘球蚴长爪沙鼠模型。随机分为模型组（无干预,40只）和实验组（抗OPN抗体干预,20只）。感染至100 d时剖杀所有长爪沙鼠,观察泡状棘球蚴生长和转移情况,取长爪沙鼠肝泡状棘球蚴组织和转移灶标本。采用免疫组织化学SP法观察沙鼠肝泡状棘球蚴组织和转移灶中OPN及IL-10的表达情况。 结果 60只长爪沙鼠全部感染肝泡状棘球蚴。免疫组化染色显示肝泡状棘球蚴组织可见OPN及IL-10不同程度的表达。模型组中OPN和IL-10阳性细胞表达率分别为72.5%（29/40）和65%（26/40）,OPN及IL-10主要分布在肝泡状棘球蚴纤维囊壁,阳性产物均定位于细胞浆。65%（26/40）的长爪沙鼠模型经病理证实发生胸廓内淋巴结转移,伴有胸廓淋巴结转移的肝泡状棘球蚴组织中OPN及IL-10阳性细胞表达率分别为84.6% (22/26)、76.9%(20/26),均高于未发生胸廓淋巴结转移的肝泡状棘球蚴组织（50%、42.9%）（P﹤0.05）。实验组OPN和IL-10的阳性细胞表达率分别为40%（8/20）和35%(7/20)。OPN与IL-10表达呈正相关（r=0.605,P=0.000）。结论 OPN阳性表达可能与肝泡状棘球蚴的转移有关,OPN与IL-10可能在肝泡状棘球蚴的生长中起协同作用。     

骨桥蛋白与肝棘球蚴病关系的研究进展

骨桥蛋白是一种带负电荷、具有亲水性的分泌型磷酸化糖蛋白,由多种组织细胞合成与分泌,存在于体内多种组织和细胞中。该蛋白结构上与多种基质蛋白相似,功能上具有细胞因子的特点,参与了机体多种病理过程。近年来的研究证实,骨桥蛋白在肝棘球蚴的外囊中大量表达,并在其形成过程中起重要作用。     

抗骨桥蛋白抗体抑制肝泡型棘球蚴侵袭性生长和转移

目的外源性给予抗骨桥蛋白(osteopontin,OPN)抗体,观察其对肝泡型棘球蚴生长和转移的抑制作用。方法采用开腹直视下肝脏穿刺接种泡型棘球蚴组织混悬液的方法(0.1 ml/只,约含原头节400个),建立肝泡型棘球蚴沙鼠模型180只,随机分为3组。实验组和对照组于接种后当天经尾静脉分别注射0.15 ml抗OPN抗体(效价1∶32)和沙鼠注射用的兔血清,共注射17次,前7次每次间隔2 d,后10次每次间隔1周。模型组未作处理。上述3组沙鼠分别于感染后1、20、40、60、80和100 d随机各处死10只沙鼠,观察泡型棘球蚴的生长和转移情况,采用免疫组织化学SP法观察各组沙鼠肝泡型棘球蚴组织中OPN的表达情况。于感染后100 d乙醚轻度麻醉沙鼠后,摘眼球取血,分离血清,用ELISA测定沙鼠血清中OPN的含量。结果感染后1、20、40、60和80 d,实验组与对照组和模型组囊湿重、胸腔淋巴结转移率间的差异均无统计学意义(P>0.05)。感染后100 d,实验组囊湿重、胸腔淋巴结转移率[(7.28±0.38)g、20%]均低于对照组[(9.70±0.61)g、70%]和模型组[(9.32±0.73)g、70%](...     

骨桥蛋白与肝泡型棘球蚴转移的相关性研究

目的研究骨桥蛋白(OPN)在肝泡型棘球蚴组织中的分布和表达,探讨骨桥蛋白在肝泡型棘球蚴转移中的作用。方法采用开腹直视下肝脏穿刺注射的方法,用20%泡型棘球蚴组织混悬液感染长爪沙鼠40只,每只0.1ml,建立肝泡型棘球蚴长爪沙鼠模型。感染100 d后剖杀所有长爪沙鼠,观察泡型棘球蚴生长和转移等情况,取长爪沙鼠肝脏、肝泡型棘球蚴组织和转移灶标本。采用苏木素-伊红(HE)染色法和免疫组织化学SP法观察沙鼠肝泡型棘球蚴组织和转移灶中骨桥蛋白的表达情况。结果感染泡型棘球蚴的长爪沙鼠的肝脏和腹腔中见大小不等的团块状囊泡。免疫组织化学染色显示,肝泡型棘球蚴长爪沙鼠模型中肝泡型棘球蚴组织中可见骨桥蛋白不同程度的表达,其阳性细胞检出率为70%(28/40),骨桥蛋白主要分布在肝泡型棘球蚴纤维囊壁、炎症细胞和部分肝细胞中。60%(24/40)的长爪沙鼠模型经病理证实发生胸廓内淋巴结转移,伴有胸廓淋巴结棘球蚴转移的肝脏泡型棘球蚴组织的OPN阳性细胞检出率(83%,20/24)高于未发生胸廓淋巴结转移的肝脏泡型棘球蚴组织(50%,8/16)(P<0.05)。淋巴转移灶中骨桥蛋白阳性细胞检出率(92%,22/24)明显高于泡球蚴组织(...     

抗OPN抗体干预感染泡球蚴沙鼠后细胞因子水平的动态变化

目的观察人工接种泡球蚴(EM)沙鼠在抗骨桥蛋白(OPN)抗体干预下体内白细胞介素-2(IL-2)、肿瘤坏死因子-α(TNF-α)、IL-5和IL-10水平动态变化,探讨抗OPN抗体对泡球蚴感染免疫机制的影响。方法采用开腹直视下肝脏穿刺注射的方法,用20%泡球蚴组织混悬液感染长爪沙鼠180只,每只0.1ml,建立肝泡球蚴病(AE)沙鼠模型。将感染沙鼠随机分为实验组、对照组和空白组,每组60只。实验组沙鼠接种泡球蚴后当天经尾静脉注射抗OPN抗体,每次0.15ml,每隔两日注射一次,注射7次后,每隔1周注射一次;对照组沙鼠经尾静脉注射兔血清(时间和剂量同上);空白组为单纯模型组。分别于感染1、20、40、60、80和100d后,各组随机选取10只沙鼠,摘眼球取血,离心,收集血清,应用酶联免疫吸附试验(ELISA)检测IL-2、TNF-α、IL-5和IL-10含量。结果感染100d时实验组IL-10为(77.93±4.13)pg/ml,对照组为(82.46±4.24)pg/ml,空白组为(84.69±5.57)pg/ml,差异有统计学意义(P<0.05),其余各时间段的IL-10水平及各时间段其他3种细胞因子水平组间比较差异无统计学意义(P>0.05)。结论 Th1细胞介导的细胞免疫与Th2细胞介导的体液免疫共同参与宿主抗肝泡球蚴免疫。抗OPN抗体可能对血清IL-10表达水平有一定的抑制作用。     

骨桥蛋白在肝细粒棘球蚴外囊壁中的表达

目的 研究骨桥蛋白（osteopontin，OPN）在肝细粒棘球蚴外囊壁中的分布及表达。 方法 用免疫组化、免疫荧光双标记法观察60例患者手术切除的肝细粒棘球蚴外囊壁及巨噬细胞中OPN的表达与分布；Von Kossa染色观察囊壁中钙化分布特征。 结果 肝细粒棘球蚴外囊壁中有不同程度OPN表达，75%（45/60）集中分布于近虫体侧纤维囊壁（内层），.3%（5/60）分布于近肝组织侧纤维性囊壁（外层），两者差异有统计学意义（P＜0.01）。在内、外层交界处可见巨噬细胞带，多数巨噬细胞胞浆内有OPN表达。OPN表达阳性的囊壁均合并有不同程度的钙盐沉积，其在囊壁内、外层的分布与OPN的基本一致。 结论 OPN主要分布在肝细粒棘球蚴外囊的内层纤维囊壁。     

肝泡状棘球蚴病

肝泡状棘球蚴病 (Ａｌｖｅｏｌａｒｅｃｈｉｎｏｃｏｃｃｏｓｉｓ ,Ａ ｅ .)是一种对人体危害极大的人畜共患的寄生虫病 ,由多房棘球绦虫的幼虫寄生于肝脏并不断增殖芽生 ,似癌样浸润扩散 ,造成肝组织不可逆的进行性损害。临床上具有慢性隐袭进行性的特点 ,常伴有肺及脑等重要器官转移。由于Ａ ｅ .具有一定的地区分布性 ,易于引起误诊 ,因此复习有关文献 ,提高Ａ ｅ .的全面认识 ,对临床诊治工作必将有所裨益。1　流行病学[1- 5]泡球蚴病主要流行于北半球高纬度地带。我国主要在西北地区流行。　　多房棘球绦虫的主要终宿主是狐及狼等野生…     

肝泡状棘球蚴病的超声及病理表现

目的: 分析肝泡状棘球蚴病患者的临床、病理及超声图像特征.方法: 回顾性分析2002-2007年我院经手术病理证实的肝泡状棘球蚴病患者15例的超声图像特征并分型.结果: 15例肝泡状棘球蚴病单发12例,多发3例,共发现肿块20个;超声特征:多位于左肝,大小多为5 cm以上,形态多不规则,边界多清晰,内部回声多呈高回声、不均匀、无液化、无钙化,后方回声多衰减,无声晕、无内部血流多见,未见肝门及腹腔淋巴结肿大;根据声像图特征分为:实性肿块型(13个,65%)、肿块液化型(7个,35%)、肿块钙化型(8个,40%).结论: 超声检查肝泡球蚴病超声影像具有特征性,是肝泡球蚴病重要的影像学检查方法.     

年龄对小鼠继发性肝泡状棘球蚴模型的影响

目的 探讨不同年龄对继发性肝泡状棘球蚴的影响。方法 分别选取8周龄、18周龄、28周龄雌性昆明小鼠29、25、25只,用20%乌拉坦腹腔注射麻醉后,运用切开上腹部皮肤经腹壁肌层透视下肝穿刺方法分别对3组小鼠进行肝脏注射E. m组织混悬液,制备继发性肝泡状棘球蚴小鼠模型。接种后饲养100 d,行安乐死并解剖。结果 3组小鼠存活率分别为: 62.1%、84%、68%(P>0.05)。肝脏E. m感染率分别为:72.2%、71.4%、76.5%(P>0.05)。肝脏E. m包囊直径分别为: 0.915±0.103 cm、1.247±0.112 cm、1.215±0.197cm(P>0.05)。肝脏E. m包囊质量分别为:0.332±0.035 g、0.532±0.155 g、0.382±0.085 g(P>0.05),差异无统计学意义,小鼠肝E. m组织HE染色无差异。结论 利用18周龄小鼠作为造模动物制作继发性肝泡状棘球蚴模型,实验小鼠存活率高。     

抗骨桥蛋白抗体对沙鼠肝多房棘球蚴组织周围血管生成的影响

目的观察抗骨桥蛋白（osteopontin,OPN）抗体对长爪沙鼠肝多房棘球蚴（俗称泡球蚴）组织周围血管生成的影响。方法 90只长爪沙鼠随机分为3组,即模型对照组（A组）,兔血清对照组（B组）和抗OPN抗体干预组（C组）,所有沙鼠均采用开腹肝穿刺法接种泡球蚴原头节悬混液0.1ml（约含原头节400个）。B组于接种当天注射兔血清0.15ml/鼠,1次/2d,连续7次,之后改为每周1次,直至处死;C组采用相同的方法注射抗OPN抗体（效价1∶32）;A组不作任何处理。分别与感染后第20、60、100、140、180d每组各处死6只沙鼠,留取肝泡球蚴组织,制作组织切片采用苏木素-伊红（H-E）法和免疫组化Envision法观察各组沙鼠肝泡球蚴组织MVD-CD34的表达情况。结果感染泡球蚴沙鼠肝脏中均可见团块状泡球蚴组织,部分播散至腹腔。感染20d时A、B、C组MVD分别为（9.83±3.87）/HP,（9.67±2.94）/HP和（7.50±1.87）/HP,感染60d时分别为（33.67±3.67）/HP,（32.83±6.11）/HP和（24.33±5.61）/HP,感染100d时分别为（44.67±4.92）/HP,（42.20±6.26）/HP和（28.00±8.76）/HP,感染140d时分别为（34.17±3.19）/HP,（31.67±4.97）/HP和（20.50±4.72）/HP;感染180d时分别为（32.33±7.42）/HP,（29.67±3.88）/HP和（13.50±3.21）/HP。其中感染60d及以后各时间点C组与A组和B组相比,微血管数差异有统计学意义（P〈0.05）。结论抗OPN抗体可抑制沙鼠肝泡球蚴组织周围血管生成。     

细粒棘球蚴和多房泡球蚴在人体内蛋白质表达谱初步分析

目的 初步掌握流行于青海省细粒棘球绦虫幼虫棘球蚴(原头节、囊壁、囊液)和多房棘球绦虫幼虫泡球蚴在中间宿主人体内蛋白质表达情况。方法 利用SDS-PAGE和 Western-blot分析棘球蚴原头节、囊壁、囊液蛋白质和泡球蚴总蛋白表达谱。结果 细粒棘球蚴原头节的蛋白质浓集在分子量72 kDa处,囊壁的蛋白质浓集在72 kDa、26 kDa和17 kDa 处,囊液的蛋白浓集在72 kDa、43~55 kDa、26 kDa和17 kDa;泡球蚴总蛋白质浓集在分子量72 kDa、55~72 kDa和26 kDa处。结论 不同地区细粒棘球蚴在人体内蛋白的表达存在差异;不同亚种泡球蚴原头节蛋白的表达存在差异。     

细粒棘球蚴和多房棘球蚴感染对Balb/c小鼠淋巴细胞亚群的影响

目的 分析Balb/c小鼠感染细粒棘球蚴和多房棘球蚴后脾脏和血液淋巴细胞亚群的变化.方法 利用流式细胞仪分析感染细粒棘球蚴和多房棘球蚴的Balb/c小鼠脾脏和血液中T、B、Th1、Th2、Treg、NK细胞亚群变化.结果 脾脏B、Th1、Th2、Treg细胞在多房棘球蚴感染组和对照组比较,差异有统计学意义;血液Th1和...     

大鼠作为泡球蚴保种动物模型的实验观察

目的研究大鼠作为泡球蚴保种模型动物的可行性。方法选取Wistar、SD大鼠,使用在长爪沙鼠体内保种的泡球蚴组织碎片进行腹腔接种,并对接种成功鼠种进行种内二次传代和抗泡球蚴组织多克隆抗体的免疫组化鉴定。同时用新西兰白兔、豚鼠、长爪沙鼠建模,观察泡球蚴感染情况。结果新西兰白兔、豚鼠、未能通过腹腔接种感染,Wistar大鼠、SD大鼠获得感染,感染率分别为85%和80%,长爪沙鼠感染率100%;感染大鼠可有一个较长的存活期,体内可见到生长活跃的泡球蚴虫体组织和原头节;泡球蚴能在Wistar大鼠、SD大鼠中稳定传代。结论 Wistar和SD大鼠是较理想的泡球蚴保虫动物。     

Radiological findings of alveolar hydatid disease of the lung caused by Echinococcus multilocularis

Alveolar hydatid disease is a highly malignant form of echinococcosis caused by the larvae of the cestode Echinococcus multilocularis. Alveolar hydatid disease always affects the liver and can metastasise to the lungs and brain. The case reports describe the radiological features of alveolar hydatid disease of the lung caused by E. multilocularis. Multiple nodules which varied in size and shape were seen on CXR, CT showed most nodules to be lobulated, well circumscribed and of varying shape. Multiple lobulated lesions located between two segments of the lung and of varying shape appear to be characteristic of pulmonary alveolar hydatid disease caused by E. multilocularis.     

Controversies in the laparoscopic treatment of hepatic hydatid disease

BackgroundLaparoscopic treatment of hydatid disease of the liver can be performed safely in selected patients.MethodsSix hundred and fifty patients were treated for hydatid disease of the liver between 1980 and 2003 at the Hepatopancreatobiliary Surgery Unit of Istanbul Medical Faculty, Istanbul University. Of these, 60 were treated laparoscopically between 1992 and 2000. A special aspirator-grinder apparatus was used for the evacuation of cyst contents. Ninety-two percent of the cysts were at stages I, II or III according to the ultrasonographic classification of Gharbi.ResultsConversion to open surgery was necessary in eight patients due to intra-abdominal adhesions or cysts in difficult locations. There was no disease- or procedure-related mortality. Most of the complications were related to cavity infections (13.5%) and external biliary fistulas (11.5%) resulting from communications between the cysts and the biliary tree. There were two recurrences in a follow-up period ranging between 3.5 and 11 years.DiscussionLaparoscopic treatment of hydatid disease of the liver is an alternative to open surgery in well-selected patients. Important steps are the evacuation of the cyst contents without spillage, sterilization of the cyst cavity with scolicidal agents and cavity management using classical surgical techniques. Our specially designed aspirator-grinder apparatus was safely used to evacuate the cyst contents without causing any spillage. Knowledge of the relationship of the cyst with the biliary tree is essential in choosing the appropriate patients for the laparoscopic technique. In our experience of 650 cases, the biliary communication rate was as high as 18%; half of these can be detected preoperatively. In the remaining, biliary communications are usually detected during or after surgery. Endoscopic retrograde cholangiopancreatography (ERCP) and sphincterotomy are helpful to overcome this problem. As hydatid disease of the liver is a benign and potentially recurrent disease, we advocate the use of conservative techniques in both laparoscopic and open operations.     

Epitope specificities and antibody responses to the EG95 hydatid vaccine

Antibody isotype and epitope specificities were examined in sheep immunized with EG95, a protective recombinant vaccine against hydatid disease. All sheep immunized with EG95 as a fusion protein with glutathione S-transferase (GST) produced prominent IgG antibodies against the EG95 portion of the protein. Linear, antibody-binding epitope specificities of EG95 were mapped using a series of 25 overlapping synthetic peptides. Three immunodominant regions were identified which generated specific IgG₁ and IgG₂ antibodies in the majority of vaccinated sheep. These regions corresponded to the EG95-derived sequences SLKAVNPSDPLVYKRQTAKF, DIETPRAGKKESTVMTSGSA and SALTSAIAGFVFSC. An additional immunogenic region was identified which induced almost exclusively IgG₂ antibody. This epitope was located within the sequence TETPLRKHFNLTPV. The anti-parasitic, protective effects of the EG95 vaccine correlated with the detection of specific antibody to two or more of the four linear immunogenic regions. The identification of these immunogenic peptides of EG95 maybe useful in the development of a synthetic peptide vaccine as a derivative of the EG95 recombinant.     

激活素A中和抗体对小鼠肝组织细胞因子mRNA表达影响

目的　应用四氯化碳(CCl4)制备小鼠肝纤维化模型,再给予激活素(ACT)A中和抗体,观察ACTA中和抗体对小鼠肝纤维化的抑制作用及对相关细胞因子表达的影响。方法　将健康雄性昆明种小白鼠分为5组:正常对照组、橄榄油对照组、CCl4 模型组、ACTA中和抗体组及对照抗体组。采用逆转录聚合酶链反应(RT -PCR)半定量法分析肝组织内ACTA、转化生长因子β1 (TGF -β1 )、基质金属蛋白酶组织抑制物1(TIMP -1)及血小板源性生长因子(PDGF)的表达。结果　与CCl4 模型组和对照抗体组相比, ACTA 中和抗体组肝纤维化病理学变化减轻,肝组织内ACTA、TGF -β1、TIMP -1 及PDGF的mRNA表达减少(P<0.01)。结论　ACTA中和抗体中和ACTA的作用,可能通过减轻对肝实质细胞的生长抑制和凋亡诱导而发挥抗肝纤维化作用,从而使肝纤维化形成的关键细胞—肝星状细胞的细胞外基质成分表达减少,进而使与之有关的细胞因子网络系统发生改变,肝纤维化程度减轻。     

Effect of antibacterial antibody on bactericidal activities of superoxide and lysosomal enzyme from alveolar macrophages in rabbits

Abstract Alveolar macrophages (AM) have many Fc receptors for IgG, but are less reactive to lymphokines. They have a well-developed oxidative metabolism and contain large amounts of lysosomal enzyme. This suggests that the antibacterial antibody plays an important role in early resistance by AM to intracellular bacterial infection and that a bactericidal agent, dependent on oxygen and lysosomal enzyme, participates in the intracellular killing of bacteria in AM. We studied the effects of the antibacterial antibody on bactericidal activities of superoxide (O₂⁻) and lysosomal enzyme from rabbit AM. The number of Listeria monocytogenes in AM increased after pretreatment with saline or normal IgG but decreased by 60% after pretreatment with anti-Listeria and 120 min incubation. Alveolar macrophage-phagocytized Listeria monocytogenes and Bacille Calmette Guérin (BCG) bound with antibacterial antibody enhanced release of O₂⁻, and nitroblue tetrazolium (NBT) formazan reduced by O₂⁻ was observed around the bacteria in the phagosomes of AM. We also confirmed that Listeria and BCG were killed extracellularly by O₂⁻ released by a superoxide-generating system in vitro and/or by lysosomal enzyme obtained from AM at a low pH. We concluded that the antibacterial antibody of the IgG class enhances the antibacterial activity of AM thereby increasing the production of O₂⁻ and lysosomal enzyme in the phagosome. This finding may be important in the early resistance to intracellular bacteria infection by AM in the alveolar spaces.