肝包虫周围纤维囊壁中骨桥蛋白免疫印记与组化分析

目的:研究肝细粒棘球蚴(肝包虫)周围纤维囊壁中骨桥蛋白(osteopontin,OPN)的表达定位．方法:采用免疫印记法检测OPN在肝细粒棘球蚴(n=48)周围纤维囊壁中的表达,并结合免疫组化方法观察其组织学分布．结果:肝细粒棘球蚴周围纤维囊壁经免疫印记分析80％有OPN表达,形态学观察集中分布于近虫体侧纤维性囊壁(内层),与近肝侧纤维性囊壁(外层)比较有显著差异(75％ vs 8．3％, P<0．05)．结论:OPN参与肝细粒棘球蚴周围内层纤维性囊壁肉芽肿反应的调节．     

肝细粒棘球蚴周围纤维囊壁钙化分布及意义

目的 研究肝细粒棘球蚴周围纤维囊壁（外囊）的钙化分布特征并探讨其意义。方法应用特殊染色观察60例肝细粒棘球蚴外囊壁病理形态特点及钙盐沉着的分布特征．并结合病人血清生化指标及CT观察结果进行分析。结果肝细粒棘球蚴外囊壁中钙盐沉着集中分布于近虫体侧内层纤维性囊壁．且形态各异．与近肝侧外层纤维性囊壁比较有显著差异（P＜0．05）；病人血清钙代谢生化指标均为常值,钙化与非钙化组比较均无显著差异（P均＞0．05）；染色可见部分CT未见钙化的囊壁有钙盐沉着。结论 肝包虫周围纤维性囊壁可分为内层和外层，且各自形成机制不同；钙盐沉积量与内层纤维囊壁形成进程有关。     

肝脾棘球蚴囊周围纤维性囊壁形成机制的差异及临床意义

目的　探讨肝、脾棘球蚴囊周围纤维性囊壁的不同形成机制。　方法　苏木素 伊红染色 ,观察 40例肝棘球蚴囊、15例脾棘球蚴囊周围纤维囊壁及其邻近肝、脾实质病理组织学改变。免疫组织化学方法检测Ⅳ型胶原、纤维连接蛋白 (FN)、层粘连蛋白 (LN) ;原位杂交方法检测转化生长因子-β1(TGF-β1)及肿瘤坏死因子-α (TNF-α)的mRNA在肝、脾棘球蚴囊周围纤维囊壁及其邻近肝、脾实质中的表达。　结果　肝棘球蚴囊周围纤维囊壁分两层 ,虫体侧纤维囊壁为肉芽肿样组织 ,肝实质侧纤维囊壁内可见大量受挤压的门静脉、肝动脉和肝管系统 (Glisson)和肝静脉系统 ,并与Glisson鞘相延续。Ⅳ型胶原、FN、LN、TGF-β1及TNF-α在两层中的表达差异均有显著性意义 (P均 <0.01)。脾棘球蚴囊周围纤维囊壁不分层 ,为肉芽肿样组织 ,Ⅳ型胶原、FN、LN、TGF-β1及TNF-α无特异分层表达。 　结论　肝、脾棘球蚴囊周围纤维囊壁的形成机制不同。肝棘球蚴囊周围纤维性囊壁是人体形成肉芽肿样病理改变后被周围受挤压的Glisson系统和肝静脉系统包裹 ,过度肝纤维化所致 ,肉芽肿样组织与周围纤维化的Glisson系统和肝静脉系统间有可分离间隙。脾棘球蚴囊周围纤维性囊壁是肉芽肿样组织包裹虫体形成 ,其与脾实质间无可分离间隙     

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。     

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。     

肝细粒棘球蚴周围纤维囊壁分层研究现状简介

肝包虫病是新疆地区常见的地方性流行性疾病。临床实践、病理组织学观察以及免疫组化等实验证实,肝细粒棘球蚴周围纤维囊壁可分为内外两层——近肝侧的"外膜"层,与近虫体侧的"外囊"层,且两层的形成机制有所不同。     

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。     

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(...     

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。     

细粒棘球绦虫在人体内棘球蚴原头节、囊壁蛋白质表达谱分析

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）在中间宿主人体内蛋白质表达情况。方法利用SDS-PAGE和western—blot分析棘球蚴原头节和囊壁蛋白质表达谱,进一步利用2DE和MALDI-TOF质谱分析技术对原头节和囊壁蛋白质构成情况进行全面的研究分析。结果在原头蚴、囊壁组织的双向电泳图中,各蛋白点通过MAL—DI—TOF—MS检测,原头蚴和囊壁分别有40个蛋白质点和8个蛋白质点得以初步鉴定,包括有：（1）细胞骨架蛋白：肌动蛋白、原肌球蛋白。（2）代谢相关酶类：苹果酸脱氢酶;磷酸丙糖异构酶;脯氨酸氧化酶;谷胱甘肽-S-转移酶;类胡萝卜脱氢酶;肽基脯氨酰顺反异构酶;（3）物质转运相关蛋白：载脂蛋白A-I;（4）应激反应蛋白（HSP70,HSP20相关蛋白）。结论初步鉴定了寄生人体细粒棘球蚴原头蚴、囊壁组织中的部分蛋白质点,其中,原头蚴中共有40个蛋白点,囊壁中有8个蛋白点。这些蛋白可能与细粒棘球蚴运动、生长、发育、代谢调控等方面有关。     

Detection of Osteopontin in the pericyst of human hepatic Echinococcus granulosus

It aims at investigating the expression and distribution of the Osteopontin (OPN) in the pericyst of human hepatic Echinococcus granulosus and their related significances. Sixty pericysts excised by “sub-adventitial cystectomy” were studied. OPN was detected in 80% (48/60) of cysts by Western blotting and distributed in the side of “exocyst” layer directing to the parasite, also macrophages were identified in the vicinity of OPN by immunohistochemistry staining. The coexpression of OPN and CD68 was observed by immunofluorescence double labeling and analyzed by Image-Pro Plus 5.1; with special stain techniques, variable degrees of calcium deposits were observed in 80% (48/60) cysts, and the calcium deposits concurrencely found with the OPN expression. The selective distribution of OPN, calcium in the “exocyst” provides a new pathological evidence for the “sub-adventitial cystectomy” we developed. The pericyst of hepatic E. granulosus consists of two detachable layers with different formative mechanisms: the “exocyst” layer directing towards the cyst of parasite was the result of granulomatous reaction; also the results suggest OPN is one regulator in the granulomatous reaction and calcification of “exocyst”.     

In-vitro susceptibility of hydatid cysts of Echinococcus granulosus to nitric oxide and the effect of the laminated layer on nitric oxide production

Murine hydatid cysts of Echinococcus granulosus were incubated in vitro in the presence of nitric oxide produced from S-nitroso-N-acetylpenicillamine (SNAP) or interferon-gamma activated peritoneal macrophages. In both situations, evidence of cyst damage and death was observed by microscopy in over 77% of cysts after 3 days, indicating that intact hydatid cysts could be susceptible to a Th1 driven macrophage attack. A crude extract of the laminated layer from cysts was found to be able to reduce the production of nitric oxide from activated macrophages in vitro and in vivo and this may have been due to phagocytosis of laminated layer fragments by the macrophages. The results indicate that, although cysts may be susceptible to the effects of nitric oxide, the laminated layer may be involved in downregulating nitric oxide production.     

Ⅰ、Ⅲ、Ⅳ型胶原在肝包虫囊肿周围人体纤维囊壁中的特异性分层表达

探讨Ⅰ、Ⅲ和Ⅳ型胶原在形成肝包虫囊肿周围人体纤维囊壁中的作用及其临床意义。采用免疫组织化学方法检测Ⅰ、Ⅲ和Ⅳ型胶原在40例肝包虫囊肿周围纤维囊壁中表达。Ⅰ、Ⅲ和Ⅳ型胶原在肝包虫囊肿周围纤维囊壁中出现特异性分层表达。靠近虫体侧纤维囊壁中,Ⅰ、Ⅲ和Ⅳ型胶原的表达阳性率分别为500%、375%和400%。靠近肝实质侧纤维囊壁中,Ⅰ、Ⅲ和Ⅳ型胶原的表达阳性率分别为875%、825%和850%。Ⅰ、Ⅲ、Ⅳ型胶原在两层中表达的差异均有显著意义(P<001,P<001,P<001)。肝包虫囊肿周围人体纤维囊壁分层,Ⅰ、Ⅲ和Ⅳ型胶原与肝实质侧纤维囊壁的形成有密切关系。     

微囊法棘球蚴继发感染小鼠动物模型的建立

目的 通过体外培养原头蚴发育成微囊,经腹腔注射建立稳定的细粒棘球蚴和多房棘球蚴继发感染小鼠动物模型。方法 无菌条件下采集羊源细粒棘球绦虫原头蚴和鼠源多房棘球绦虫原头蚴,经胃蛋白酶消化后检测虫体活力并计数,于37℃、5%CO₂条件下进行体外培养至发育成微囊,以每鼠50个微囊的剂量经腹腔注射的途径分别接种BALB/c小鼠。接种6个月后,通过腹部解剖大体观察和病理检测分析各组小鼠的感染情况及包虫囊的生长情况。结果 原头蚴在体外培养60d时发育成微囊,显微镜下观察Eg具有明显的透明角质层结构,而Em微囊角质层较薄。小鼠细粒棘球蚴和多房棘球蚴的感染率均为100%,Eg包虫囊为游离单囊,成囊率达70%,囊内无原头蚴;Em包虫囊为类似肿瘤的团块状组织,病灶内有生发囊及原头蚴。结论 采用微囊法可建立稳定的棘球蚴继发感染小鼠动物模型,为疫苗研制、药物筛选和疗效判定提供研究动物模型。     

The possible role of the age of the human host in determining the localization of hydatid cysts

Cystic hydatid disease or cystic echinococcosis (CE) is a cyclozoonotic infection distributed world-wide. The morbidity attributable to the infection depends on the size of the cyst(s) and the organ(s) involved. The cysts are most commonly found in the liver and lungs but certain locations have been reported to be more prevalent in children and/or young adults than in older subjects. In order to identify the relationship, if any, between the age of the patient and the site of involvement, the age and cyst distribution of 92 cases of CE were analysed. Lung, brain, spinal and orbital hydatid cysts were more commonly seen in younger patients whereas other sites were preferentially involved in older patients. The factors that determine the final localization of the cysts are discussed. It is concluded that age somehow alters the host-parasite relationship and thus affects the organ distribution of the cysts.     

超声造影剂对高强度聚焦超声破坏棘球蚴囊壁的增强作用

目的 观察高强度聚焦超声(high intensity focused ultrasound,HIFU) 联用超声造影剂(ultrasound contrast agent,UCA)辐照细粒棘球蚴包囊后囊壁的损伤情况。方法 15个直径为30 mm左右的包囊随机分为3组:对照组,接受超声假照;单纯HIFU组,接受100 W HIFU辐照;HIFU+UCA组,注入0.1 mL UCA后进行HIFU辐照。辐照后观察超声图像的灰度变化和囊壁的肉眼变化,并取囊壁组织制作病理切片、扫描电镜切片和透射电镜切片,观察其病理改变。结果 100 W HIFU辐照可破坏囊壁的部分生发层和角质层;加入UCA后进行100 W HIFU辐照,生发层几乎无残留,角质层也遭到更严重的破坏。结论 UCA能增强HIFU对离体细粒棘球蚴囊壁的破坏作用。     

A retrospective study on the coexistence of hydatid cyst and aspergillosis.

OBJECTIVES: Hydatid cyst is a zoonotic disease with an endemic regional distribution, and Aspergillus is a saprophytic fungus that may cause allergic pulmonary aspergillosis, aspergilloma, and semi-invasive and invasive aspergillosis. The coexistence of a saprophytic fungus and hydatid cyst is extremely rare. The aim of this retrospective study was to evaluate the coexistence of aspergillosis and echinococcosis in archival materials and to discuss its probable clinical significance. METHODS: Hematoxylin-eosin (HE)-stained sections of 100 archival cases with the diagnosis of hydatid cyst were reevaluated by four pathologists independently. Grocott's methenamine-silver (GMS) and periodic acid-Schiff (PAS) were applied to the slides that were suspected of having co-infection with Aspergillus to confirm the diagnosis. RESULTS: Two cases of aspergillosis and hydatid cyst coexistence were found out of the 100 reevaluated archival cases with a diagnosis of hydatid cyst. Both of the cases were located in the lung, in immunocompetent patients. CONCLUSIONS: Aspergillosis and hydatid cyst coexistence may be important in patients with immune deficiency and in cases with pre- or perioperatively ruptured cysts. There are no reliable data on the specificity and sensitivity of radiological imaging techniques in detecting the existence of Aspergillus in hydatid cysts. Histopathological evaluation is essential for diagnosis and for the planning of management.