肝泡状棘球蚴病不同手术疗效及预后的分析研究

目的:比较肝泡状棘球蚴病(HAE)不同手术方式的疗效,并分析影响HAE手术患者预后的相关因素。方法:回顾性分析2003年9月-2015年2月期间在我院进行手术治疗的HAE病人的诊疗记录。根据手术方式的不同,将病人分为非移植性根治性切除组(A组)、术中病灶绝大部分切除(90%以上)组(B组)、术中不能90%以上切除或仅引流组(C组)、肝移植组(D组)。结合随访资料,评价四组的疗效,并分析影响患者预后的相关因素。结果:A组死亡率低于其他三组,差异具有统计学意义(P0.001)。生存曲线结果显示,A组预后生存状况优于其他三组,差异具有统计学意义(P=0.001)。多因素分析结果表明,非移植性根治性切除、术中出血量是影响患者生存的独立危险因素(均P0.05)。结论:在早期发现早期诊断的前提下,对HAE病人行非移植性根治性切除术治疗效果最好,且非移植性根治性切除是患者预后的独立危险因素。     

内蒙古西伯利亚棘球绦虫和多房棘球绦虫泡状蚴在小白鼠发育成熟的比较

The alveolar echinococcus is one of the most dangerous worm parasites in man. Rausch and Schiller reported a new species, Echinococcus sibiricensis n. sp. from arctic fox, Alpex logopus, on St. Lawrence Island of Alaska, USA. According to the view of Vogel, the sibiricensis form is only a geographical race or subspecies of Europe Echinococcus multilocularis. So far, the two names, Echinococcus multiocularis multilocularis and Echinococcus multilocularis sibiricensis, existed in many references and text books. We have found the adults of Echinococcus sibiricensis and Echinococcus multilocularis from sand foxes, Vulpes corsac and their larval stages (alveolar echinococcus) from field voles, Microtus brandti in the Hulunbeier Pasture of Inner Mongolia, northeastern China in 1985 and 1998-1999. Two types of metacestodes with quite different styles of early development of E. sibiricensis and E. multilocularis were found from field voles and laboratory experimental white mice. As one characteristic of alveolar E. multilocularis, the capsules are produced by the exogenous budding of germinal cell layer together with cyst wall. The protoscoleces grow from germinal cells on germinal cell layer. The peduncles of early protoscoleces attached to the germinal cell layer on the inner surface of capsule wall(Plate I, Figs. 1-2). Some protoscoleces in reticular structure were linked with the inner surface of capsule wall (Plate I, Fig. 3) in livers of mice in 9.5th month postinfection. In 14th month old alveolar multilocularis, large number of mature protoscoleces in reticular structure were still linked to the inner surface of capsule wall (Plate I, Figs. 4-8). The cavities of some capsules were filled with protoscoleces in meshes of reticular structure which were also linked around with the inner surface of capsule wall (Plate I, Fig. 9). The superficial surface of livers of positive field voles and experimental mice never showed any hyperemic phenomenon. The superficial surfaces of livers and lungs of positive field voles and experimental mice infected with alveolar E. sibiricensis were highly hyperemic. The metacestodes of E. sibiricensis composed of mother cyst, undifferentiated embryonic cysts and small brood capsules. Cavities of all cysts were fully filled with germinal cell masses. Host reaction appeared to be very strong, all cysts were surrounded by thick connective tissue and dense leukocytes (Plate II, Fig. 10). All alveolar vesicles were found located in lungs tissue of experimental mice. Large germinal cell masses metastasized out from undifferentiated embryonic cysts into host lung tissue, where germinal cell masses developed into accumulation of early protoscoleces (Plate II, Figs. 11-12). Early protoscoleces of alveolar E. sibiricensis were seen earliest in mice lung tissues on 101-104th days after infection. Many small capsules in different sizes and different shapes containing mature protoscoleces and reticular structure (Plate II, Figs. 13-15) were found in lungs of mice in 9th month after infection. Only in one experimental mouse infected with alveolar E. sibiricensis in 8.5th month postinfection, both its lung and liver existed alveolar cysts; the capsules in liver were surrounded by very thick connective tissue of the host, and there were some protoscoleces in their cavities (Plate II, Figs. 16-18).     

内蒙古西伯利亚棘球绦虫和多房棘球绦虫泡状蚴在小白鼠发育成熟的比较

本文报道人兽共患的泡状肝包虫病原,西伯利亚棘球绦虫Echinococcus sibiricensis (Rausch andSchiller, 1954)泡状蚴和多房棘球绦虫Ech inococcusmultilocularis ( Leuckart, 1863)泡状蚴在KM株小白鼠发育成熟过程比较观察的结果.此两虫种泡状蚴的发育成熟过程仍然和它们早期发育的规律(唐崇惕等,2001)相同.虽然它们成熟的泡囊都被着生在网状结构中的许多原头节所充满,但是在多房棘球绦虫9-14个月的泡状蚴,仍然可以见到它们的原头节和网状结构都是起源于泡囊囊壁内表面的胚细胞层,并且始终保持与该层的联系.而西伯利亚棘球绦虫泡状蚴在鼠肺脏或肝脏的各泡囊中的原头节和网状结构是由可移动的胚细胞团发育生成,它们与泡囊囊壁没有如前者样的联系.西伯利亚棘球泡状蚴在各别小白鼠肝脏也能发育成熟,但不正常,宿主反应异常强烈.     

细粒棘球绦虫原头蚴组织化学的观察

本文对细粒棘球绦虫原头蚴的组织化学成份进行了较为系统和全面的观察,结果表明,该虫体内存在糖原、RNA、DNA、蛋白质结合的α-氨基、酪氨酸、色氨酸及组氨酸、碱性蛋白质、酸性粘多糖、胶原及网状纤维成分、AKP、ATP酶及ACP等生化物质。并对上述成分的定位,含量及其生理学的意义进行了重点讨论。     

细粒棘球蚴FABP基因的原核表达及蛋白鉴定

构建原核表达载体pET-FABP,优化表达条件,采用免疫学方法鉴定纯化的FABP融合蛋白。载体pMD19-T-FABP和pET-44a(+)经EcoRⅠ和HindⅢ双酶切,将回收的FABP片段与pET-44a(+)连接,构建原核表达载体pET-FABP并在大肠杆菌BL21(DE3)中表达,优化表达条件,纯化FABP融合蛋白,Western blotting鉴定。成功构建了原核表达载体pET-FABP并在大肠杆菌中高效表达。纯化后的蛋白经SDS-PAGE和Western blotting鉴定正确。构建的表达载体pET-FABP可以在大肠杆菌中大量表达Nus-FABP融合蛋白,为进一步研制FABP亚单位疫苗奠定了基础。     

细粒棘球蚴eg95基因的克隆以及原核融合表达

从青海省羊源细粒棘球蚴提取基因组DNA,PCR扩增eg95基因并且克隆。测序结果表明该基因序列全长1262bp,由2个内含子和3个外显子组成,3个外显子分别为70bp、306bp和95bp。因此,推测其ORF为471bp,编码156个氨基酸。用RT—PCK方法扩增该基因的ORF,经克隆、DNAstar比对分析序列,它与预测的ORF序列100％相同。将目的基因克隆到原核表达载体pGEX-5x-1,构建重组表达质粒pGEX-5x-1-eg95,将其转入表达菌株BL21中进行原核表达。表达产物经过SDS—PAGE电泳分析,得到一条特异的蛋白条带;质谱鉴定证明表达的蛋白确实是eg95抗原蛋白;酶联接免疫吸附剂测定表明抗原抗体发生了特异性反应,进一步证明融合蛋白进行了正确表达,从而为囊型包虫病的研究奠定基础。     

棘球蚴囊液对小鼠脾细胞IL-17及Smad2表达的影响

目的:观察细粒棘球蚴囊液对体外培养BALB/c小鼠脾细胞白介素(IL)-17和Smad2基因表达的影响.方法:制备小鼠脾细胞悬液,接种于48孔培养板中进行培养,以不加任何干预为对照组,囊液处理组为实验组.定时取样提取总RNA,经反转录成cDNA后用实时荧光定量PCR(qRT-PCR)检测IL-17和Smad2基因的表达.结果:小鼠脾细胞IL-17表达在囊液处理6h和12h后升高(1.000±0.207 VS 3.672±0.746和1.000±0.154 VS 5.525±0.843),差异有统计学意义(P＜0.05);小鼠脾细胞Smad2表达在培养1h后升高(1.000±0.077 VS 2.069±0.098)在12h后降低(1.000±0.110 VS 0.247±0.011),差异有统计学意义(P＜0.05);协同分析发现IL-17基因与Smad2基因的表达变化呈现负相关(P＜0.01),相关系数r=-1.结论:细粒棘球蚴囊液对小鼠脾细胞IL-17和Smad2基因的表达具有上调作用,推测其可能与宿主抗棘球蚴感染的机制有关.     

细粒棘球蚴AgB8/2基因的原核表达及免疫学鉴定

旨在克隆细粒棘球蚴 AgB8/2基因,优化原核表达体系,鉴定纯化蛋白并初步确定其诊断价值.采用 RT-PCR扩增 AgB8/2基因 cDNA,构建原核表达载体pET-AgB8/2并在大肠杆菌 BL21(DE3)中表达,优化表达条件.纯化的AgB8/2融合蛋白经 Western blot鉴定正确后,采用斑点免疫金渗滤法初步验证其血清学诊断价值.结果显示,克隆的 AgB8/2基因包含了 273 bp的完整ORF,序列分析表明与其它已报道的AgB8/2 cDNA序列的同源性在 99%～100%之间.优化的表达条件为,当菌液 OD600值为0.6时,加入终浓度为0.05 mmol/L的 IPTG,37℃,振荡培养5 h.纯化的蛋白经 Western blot鉴定为目的蛋白.克隆的 AgB8/2基因高度保守,原核表达载体pET-AgB8/2构建正确并高效表达可溶性融合蛋白,可作为特异性抗原应用于斑点免疫金渗滤法检测血清抗体.     

包虫病防治研究进展

包虫病(又称棘球蚴病)是一种具有地方流行性和自然疫源性的人畜共患性寄生虫病,严重危害广大牧区的人民身体健康并阻碍畜牧业经济的发展.近年来,关于包虫病化疗药物的研究取得了很大进展;免疫预防是防止包虫病流行的比较理想的途径;利用现代分子生物学技术对棘球蚴绦虫的有效免疫原成份进行筛选和克隆,制备基因工程疫苗,为包虫病的免疫预防和免疫诊断开辟了新途径.本文就近年来各国对本病的防治概况作如下综述及展望,并简要分析其优势与意义.     

犬恶丝虫病病原生物学研究进展

从虫体的分类、分布、宿主、发育生物学和内共生体等几个方面综述了近年来在犬恶丝虫病病原生物学研究上取得成果,并提出了研究该病病原的现实意义和今后研究的主要方向.     

A Case of Budd-Chiari Syndrome Associated with Alveolar Echinococcosis

Although alveolar echinococcosis (AE) can cause a serious disease with high mortality and morbidity similar to malign neoplasms. A 62-year-old woman admitted to a hospital located in Sivas, Turkey, with the complaints of fatigue and right upper abdominal pain. On contrast abdominal CT, a 54×70×45 mm sized cystic lesion was detected in the left lobe of the liver that was seen to extend to the posterior mediastinum and invade the diaphragm, esophagus, and pericardium. The cystic lesion was seen to be occluding the inferior vena cava and left hepatic vein at the level where the hepatic veins poured into the inferior vena cava. Bilateral pleural effusion was also detected. We discussed this secondary Budd-Chiari Syndrome (BCS) case, resulting from the AE occlusion of the left hepatic vein and inferior vena cava, in light of the information in literature.     

Effect of Antioxidants on Ammonia Induced Cns-Renal Pathobiology in Sheep

The effect of antioxidants on ammonia induced CNS and renal pathobiology in 10 sheep infested by L. Cuprina larvae was investigated. The condition produces severe dermatitis, proliferation of macro-phages and hyperammonaemia, and free radicals may therefore be involved in the pathogenesis. Five of the sheep (treated group) were given daily intramuscular (im) injections of 2 g sodium ascorbate, 5.9 g dl-alpha tocopherol (11 days) and 3 g desferrioxamine mesylate (6 days) with 70 mg oral butylated-hydroxyanisole (11 days). The treatment prevented rises in jugular ammonia, creatinine, urea, sodium and pH, and decreases in water intake, urine output and glucose. The findings showed that antioxidants prevented ammonia induced CNS and renal pathobiology and suggest that free radicals contribute to the pathogenesis of the condition.     

14-3-3 proteins in Echinococcus: their role and potential as protective antigens

14-3-3 Proteins are a family of highly conserved proteins among all eukaryotic organisms studied so far. As basically intracellular proteins, they play a key role in basic cellular events related to cellular proliferation, including signal transduction, cell-cycle control, cell differentiation and cell survival. The 14-3-3 proteins have been described and characterized in several parasites, and mostly studied in Echinocuccus granulosus and Echinocuccus multilocularis. Here, we review the discoveries regarding this protein family in the genus Echinococcus, describing new data about specific aspects related with their implication in the parasite biology and immunology in the frame of the host-parasite relationship.     

Echinococcosis in China,a Review of the Epidemiology of <Emphasis Type="Italic">Echinococcus</Emphasis> spp.

Cystic echinococcosis (CE) and alveolar echinococcosis (AE) are highly significant infectious diseases occurring worldwide and caused by metacestodes of tapeworms Echinococcus granulosus and E. multilocularis, respectively. Both human CE and AE have highest prevalence rates in western and northwestern China. Livestock is the main intermediate host of E. granulosus, and wild small mammal are the main intermediate hosts of E. multilocularis. Since they range freely in pastoral areas, prey on wild small mammals and offal of livestock after slaughter, and have close relationships with humans, domestic dogs are the most important definitive host of both Echinococcus spp. with the highest risk of transmitting CE and AE to humans. Pastoralism is the occupation with the highest risk of being infected with the both kinds of echinococcosis due to the proximity of livestock, dogs, and wildlife host species. In this review, we summarize the epidemiology of human echinococcosis, the situation of parasite transmission in animal hosts, and possible transmission patterns in China. In addition, human activities and their potential influence on the transmission of echinococcosis are also discussed.     

Efficacy of Albendazole-Chitosan Microsphere-based Treatment for Alveolar Echinococcosis in Mice

This study aimed to investigate the pharmacology and anti-parasitic efficacy of albendazole–chitosan microspheres (ABZ-CS-MPs) for established intraperitoneal infections of Echinococcus multilocularis metacestodes in an experimental murine model. Male outbred Kunming mice infected with E. multilocularis Metacestodes were administered with three ABZ formulations, namely, ABZ-CS-MPs, Liposome–Albendazole (L-ABZ), and albendazole tablet (ABZ-T). Each of the ABZ formulations was given orally at three different doses of 37.5, 75, and 150mg/kg, three times a week for 12 weeks postinfection. After administering the drugs, we monitored the pharmacological performance and anti-parasitic efficacy of ABZ-CS-MPs compared with L-ABZ, and ABZ-T treated mice. ABZ-CS-MPs reduced the weight of tissues containing E. multilocularis metacestodes most effectively compared with the ABZ-T group and untreated controls. Metacestode grown was Highly suppressed during treatment with ABZ-CS-MPs. Significantly higher plasma levels of ABZ metabolites were measured in mice treated with ABZ-CS-MPs or L-ABZ compared with ABZ-T. In particular, enhanced ABZ-sulfoxide concentration profiles were observed in the mice given 150mg/kg of ABZ-CS-MPs, but not in the mice treated with L-ABZ. Histological examination showed that damages caused disorganization of both the germinal and laminated layers of liver hyatid cysts, demolishing their characteristic structures after treatment with ABZ-CS-MPs or L-ABZ. Over time, ABZ-CS-MPs treatment induced a shift from Th2-dominant to Th1-dominant immune response. CS-MPs As a new carrier exhibited improved absorption and increased bioavailability of ABZ in the treatment of E. multilocularis infections in mice.     

中国大鲵繁殖生物学研究进展

中国大鲵（Amndrias davidiamus Blanchard）是我国特有濒危的两栖物种,是研究生物进化、生物多样性、性别决定分子机制等的好材料,对于科学研究具有巨大的潜在贡献。近年来人们对它的研究力度不断加大,本文综述厂大鲵的繁殖生物学的近期研究工作,也简要探讨了大鲵繁殖生物学今后研究的主要工作,以期为大鲵繁殖生物学及繁殖技术的进一步的研究提供有价值的参考。     

Experimental studies on Echinococcus multilocularis in Japan, focusing on biohazardous stages of the parasite

Alveolar echinococcosis (AE), caused by the active growth of larval Echinococcus multilocularis mostly in the liver, is usually fatal zoonotic disease if not adequately treated. Humans become infected via oral ingestion of the parasite eggs, which are thus biohazardous to humans and should be handled under restricted conditions. In this review, we present findings in experimental studies mainly performed at a safety facility in Japan, examining the biohazadous stages of the parasite (Hokkaido isolate) including its egg and adult worm stages. This article deals mainly with the parasite development in various experimental and wild animals, environmental factors affecting viability of the parasite eggs, and molecular biological studies on adult worms. The findings shown herein have provided a basis to better understand basic biology and natural transmission of E. multilocularis in Hokkaido, a highly endemic area of AE in northern Japan, and also to establish effective preventive measures against the disease.     

An Imported Case of Cystic Echinococcosis in the Liver

A 25-year-old Uzbek male presented with right upper abdominal pain for 20 days. On radiologic studies, a huge cystic mass was noticed in the right liver which was suspected as parasitic. The patient received right hepatic segmentectomy (segment 7), and the surgically resected mass was confirmed as cystic echinococcosis (CE), measuring 10.5 cm in its diameter. The inner surface of the cyst was bile-stained. The patient was discharged on the 8th hospital day, and was rechecked 6 months after the surgical intervention without any evidence of recurrence. The present report describes findings of an imported case of CE which represented ultrasound images of the ''ball of wool''.