Vertical Transmission of Schistosoma Japonicum in the Rabbit

Human schistosomiasis japonica has long beenrecognized as a major public health concern inAsian countries including China,with millions ofpeople affected[1] . The disease is caused by thezoonotic blood fluke,S.japonicum.More than40mammals,including the rabbit,act as naturaldefinitive hosts[2 ] .Although the major route of in-fection with S.japonicum is through skin contactwith infested water,verticaltransmission,i.e.theprocess of transplacental passage ofschistosomulaefrom the infected mothe…     

Vertical Transmission of Schistosoma Jkaponicum in the Rabbit

The aim of the present study was to confirm observations on the vertical transmission of Schistosoma japonicum in the rabbit.S.japonicum-infected pregnant rabbits were used in this study.Perfusion of mother rabbits was done 9 weeks after infection in order to obtain worm burdens in relation to their initial cercarial dose.Anti-schistosoma specific IgM antibodies in serum samples collected from rabbit kittens were detected by ELISA.Our results showed that gestation period lasted the normal 29-31 days.All the exposed mother rabbits became infected with S.japonicum.Positive IgM antibody OD values were detected in 12 out of the 60 kittens examined (20.0%).In group C and A,40.0% and 17.9% of the kitten were congenitally infected,respectively.18.1% of the kittens born to mothers infected with a single dose of 200 cercariae perrabbit were positives;this is not significantly different from that obtained for the 600 dose group (22.2%).Three randomly selected IgM^ kittens harbored between one and two adult worms.The livers of these kittens displayed granulomatous lesions.It is concluded that congenital S.japonicum infection does occur in the rabbit and is affected by the mother stage of pregnancy and to a lesser extent by its infection load.     

Congenital Transmission of Schistosoma japonicumin the Rabbit

Fourteen pregnant rabbits were each infected with 300 cercariae of Schistosoma japonicum and divided into two groups.Group M (n =8)was infected during mid-gestation (the organogenetic stage)and group L (n=6)was infected during late-gestation (the post-organogenetic stage).Mother rabbits and rabbit kittens were killed 45-60 days after infection and perfused in order to obtain worm counts.Furthermore,faecal egg counts and tissue egg counts from livers were obtained from the mother rabbits as well as the rabbit kittens.All mother rabbits became infected harbouring 207.6 20.2 and 220.0 27.5 adult worms in group M and L,respectively.In groups M and L, 13.5%and 46.7% of the kittens were infected,respectively,In 12 of 14 litters at least one kitten was infected.Tne infected kittens harboured between one and three adult S.japonicum.The livers of the kittens infected with a worm pair displaced lesions,as a result of egg deposition.The results, therefore,show that congenital transmission of S.japonicum can occur in rabbits.The close anatomical resemblance between the rabbit and human placenta may be indicative of the presence of congenital transmssion of S.japomcum infection in humans.     

Congenital infection of rabbits with Schistosoma japonicum and protective immunity of offspring

Background Recently congenital infection with Schistosoma japonicum (S. japonicum) has been domonstrated in pigs, rabbits, mice and dogs. We explored the rabbit as an animal model for the congenital infection of schistosomiasis japonica and assessed the effect of a congenital S. japonicum infection on the resistance of rabbit kittens to a postnatal challenge infection.Methods Sixteen pregnant New Zealand white rabbits were infected with a single dose of S. japonicum cercariae. The exposed animals were divided into three groups according to the gestation age at the time of infection. Diagnosis of prenatally acquired S. japonicum infection in the rabbit kittens was primarily based on serological tests in combination with parasitological and histopathological findings. Congenitally infected kittens were challenged percutaneously with 100 S. japonicum cercariae to assess the effect of a congenital S. japonicum infection on kitten resistance to a postnatal challenge infection.Results The overall prevalence of congenital infection in offspring of infected mothers was 20% (12/60). The congenital infection rate in group L (late gestation) was much higher than in group E (early gestation) and group M (mid-gestation) (P<0.05). After a postnatal challenge infection, prenatally infected kittens had a 54.66% worm reduction rate, 41.45% egg reduction rate, and 51.76% granuloma size reduction rate compared to nave kittens.Conclusions This study demonstrates the possibility of congenital infection of S. japonicum in rabbits and the resistance of congenitally infected kittens to a postnatal challenge infection. These results have important implications not only for epidemiological investigations, but also in designing government control programs for schistosomiasis.     

一氧化氮与血吸虫病肝损伤发病机制的关系研究

为了研究一氧化氮在血吸虫病肝损伤发病中的作用，采用Ｓ－Ｐ免疫组织化学法，观察家兔（ｎ＝４０）经皮肤感染血吸虫尾蚴后８、１２、１６、２０、２４周家兔肝诱导型一氧化氮合成酶（ｉＮＯＳ）和巨噬细胞（ＣＤ６８）的免疫组织化学表达与定位。结果显示，血吸虫病家兔肝的虫卵肉芽周围、门静脉分支管壁、肝窦壁、中央静脉壁有ｉＮＯＳ和ＣＤ６８阳性免疫反应物，肝细胞内有ｉＮＯＳ阳性免疫反应物。正常家兔肝脏ｉＮＯＳ和ＣＤ６８免疫组织化学染色均为阴性。认为一氧化氮在血吸虫肉芽形成及肝脏微循环障碍中具有重要作用     

旋毛虫感染早期兔血清中IgM的检测

目的探讨斑，点酶联免疫吸附试验（Dot-ELISA）检测旋毛虫感染早期免血清中特异性抗体IgM在免疫诊断上的价值。方法正常家免经口灌入旋毛虫肌肉期幼虫，于感染后d7和d14静脉系血分离血清，以Dot－ELISA法检测血清中的IgM。结果感染后d7和d14旋毛虫特异性抗体IgM的阳性率分别为85．7％和100％，最高滴度可达1：6400，且与日本血吸虫和弓形虫无交叉反应。结论Dot－ELIS法检测旋毛虫特异性抗体IgM对旋毛虫感染早期诊断具有特异性较强、敏感性高、重现性好的优点．     

日本血吸虫不同发育阶段与旋毛虫抗原交叉性的研究

运用ＥＩＴＢ技术分析日本血吸虫（Ｓｊ）尾蚴、肝期童虫及雌雄成虫与旋毛虫（Ｔｓ）肌蚴抗原的交叉性。结果显示：Ｓｊ尾蚴、肝期童虫、成虫各阶段不同程度地被抗－ＴｓＳＡＲＳ，ＴｓＩＲＳ识别，其识别的抗原分子量在１５￣１００（尤５０￣７０）ｋＤ之间，两种抗血清识别的带型基本相同。所识别的各期Ｓｊ抗原中以尾蚴抗原最多，次为肝期童虫。３０天成虫仅雌虫抗原与上述两种抗血清在９７ｋＤ处显出一条弱带。抗－Ｓｊ♂ＡＲＳ     

巢式PCR法检测日本血吸虫低感染度宿主血清DNA的研究

目的探讨日本血吸虫低感染度宿主血清的DNA检测方法,为血吸虫病的早期诊断及疗效考核提供实验依据。方法建立并优化巢式PCR反应体系用于检测不同感染度家兔血清中的日本血吸虫特异性基因片段。结果在仅存活3～5对成虫的感染后家兔血清中均能扩增到特异性目的条带,尤其是感染后3d即可检出。结论巢式PCR可用于日本血吸虫低感染度宿主血清DNA检测,具有潜在应用前景。     

日本血吸虫表皮膜蛋白的研究 Ⅰ.细胞免疫和体液免疫反应观察

本文报道以血吸虫表皮膜蛋白作抗原,采用淋巴细胞转化试验和ELISA方法对感染血吸虫的家免作纵向免疫反应观察,发现感染后25天淋巴细胞转化反应达高峰,40天后逐渐下降,而IgG抗体于感染后25天开始升高并维持在一个高水平,用ELIsA方法检测10例血吸虫病人均呈阳性反应,对5例正常人及35例其他寄生虫感染者血清均为阴性反应,提出表皮膜蛋白有可能用于血吸虫病的免疫诊断。     

日本血吸虫主要虫卵抗原Sjp40在感染新西兰白兔肝脏中的表达与免疫定位

目的观察Sjp40在感染新西兰白兔肝脏沉积虫卵及其肉芽肿病变形成中的表达情况,并进行免疫定位。方法日本血吸
虫尾蚴感染新西兰白兔,收集未感染组、29 dpi组和45 dpi组肝脏,Trizol法提取各组肝脏总RNA,以日本血吸虫甘油醛-3-磷酸
脱氢酶（GAPDH）为内参,Taqman探针qRT-PCR检测Sjp40 mRNA的表达;提取各组肝脏总蛋白,以硫酸铵盐析法和Protein G
免疫亲和柱层析法纯化的抗Sjp40-McAb 9G7和抗弓形虫tSAG1-McAb Y3A8（对照抗体）,western blot检测Sjp40蛋白的表达;
制备肝脏石蜡切片,HE染色观察肝脏虫卵肉芽肿的形成与发展,进一步以免疫荧光技术进行Sjp40在肝脏沉积虫卵及其肉芽肿
组织中的定位。结果日本血吸虫感染兔呈急性肝肉芽肿病变期的45 dpi肝脏沉积虫卵中Sjp40 mRNA水平显著高于29 dpi 尚
未形成虫卵肉芽肿结节的肝脏沉积的未成熟虫卵（P<0.05）,westernb blot 证实了Sjp40 蛋白在29 dpi 和45 dpi 兔肝脏中的表
达。免疫荧光显示：Sjp40在29 dpi兔肝脏中定位于未成熟虫卵内,而45 dpi可见感染兔肝脏中沉积大量成簇内含毛蚴的成熟虫
卵及其周围肉芽肿组织均有明显荧光。结论Sjp40在日本血吸虫感染兔肝脏沉积虫卵中的转录水平随虫卵发育成熟而显著增
加,在感染兔急性肉芽肿病变期（45dpi）呈现由虫卵向其周围肉芽肿组织扩散现象,提示该分子在血吸虫肉芽肿形成与发展过
程中具有重要作用。
     

PCR对弓形虫感染家兔血液中虫体DNA的动态检测

目的了解弓形虫感染家兔血液中是否有虫体存在,探明虫体在血液中动态变化情况。方法用不同剂量的RH株弓形虫速殖子经腹腔感染家兔6只。在家兔感染前后分别采集血液,抗凝处理后,-40℃冻存备用,用普通PCR检测弓形虫感染家兔血液中弓形虫DNA。结果5只家兔在感染后的6~14d死亡,仅1只家兔在感染后的6~72d,用PCR检测手段在血液中均可检测到弓形虫DNA。结论弓形虫感染雄兔血液中有虫体存在。     

日本血吸虫感染兔肝肌成纤维细胞的动态变化及其意义

目的观察血吸虫病兔肝纤维化形成过程中肌成纤维细胞的动态变化,探讨肝纤维化发病机理。方法随机选取60只新西兰兔,每只感染日本血吸虫尾蚴（80±1）条,于感染后不同时间取其肝脏组织做成常规石蜡块,再分别进行HE染色、苦味酸天狼红染色及α－SMA免疫组织化学染色,然后观察肉芽肿大小动态变化情况,半定量分析肝纤维化程度及α－SMA阳性细胞即肌成纤维细胞表达情况;在感染后第28周用吡喹酮彻底杀虫治疗后进行γ-干扰素抗肝纤维化治疗,对照组用注射生理盐水。结果感染后第6周出现肉芽肿,大小为（13±6）×104μm2,第8周到达高峰,为（20±9）×104μm2,其后随时间的推移肉芽肿逐渐缩小,至感染后第24周后不再有明显变化（P＞0．05）,而肝纤维化程度逐渐加重;α－SMA阳性细胞于炎症早期在汇管区已有表达,为（1．1±0．3）分,炎症后期在病灶区及肝窦均有大量表达,为（7．3±1．5）分;感染后第28周生理盐水组肝纤维化程度达（5．3±1．9）分,模型组为（7．0±1．7）分,干扰素组则为（2．8±1．0）分。结论肝肌成纤维细胞是血吸虫病肝纤维化形成及发展的关键细胞。     

用ELISA考核吡喹酮治疗家兔血吸虫病疗效的实验研究

以吡喹酮治疗实验性日本血吸虫病家兔,观察治后一年内ELISA 所测得的血清抗体水平的变化,以验证其是否可作为考核疗效的方法。结果表明:凡试验结束时解剖检虫阴性的病兔,ELISA 皆于治后8～12月内阴转,仅有雄虫 1条的病兔,亦于同期内阴转;凡检出雌雄成虫的病兔,ELISA 呈持续阳性。     

血吸虫病兔血清5-羟色胺同抗体关系的研究

10只家兔随机分成两组,重感染组每只感染日本血吸虫尾蚴1200条,轻感染组感染日本血吸虫尾蚴200条,观察两组家兔血清5-羟色胺(5-HT)、5-羟吲哚乙酸(5-HIAA)同抗体的关系。发现:感染后第15d 两组家兔血清5-HT 含量均增高,且抗体高的重感染组5-HT 增高更多;5-HIAA 亦随抗体增高而     

应用PcAb—ABC—Dot—ELISA检测弓形虫CAg的研究

检测宿主体内的CAg可以早期诊断寄生虫感染,鉴于应用PcAb-ABC-Dot-ELISA检测弓形虫CAg国内尚未见报道,作者用生物素化抗弓形虫PcAb为探针,检测了15份人工感染弓形虫的兔血清,结果15份血清的CAg均呈阳性反应。用35份正常兔血清,10份自然感染球虫的兔血清及血吸虫、华校睾吸虫感染的兔血清各5份作对照,除一份正常兔血清及一份血吸虫感染的兔血清呈阳性反应外,其余均为阴性。结果表明,该法有较好的特异性及重复性。     

重组Calpain蛋白诱导抗体产生及检测日本血吸虫感染的研究

目的 研究在大肠杆菌中表达的日本血吸虫Calpain蛋白的免疫原性及其在日本血吸虫病诊断上的应用。方 法用重组Calpain蛋白免疫BALB／c小鼠，ELISA法测定特异性IgG抗体的动态变化及其IgG亚型(IgG1，IgG2a，IgG2b，IgG3)产生特征；同时用重组Calpain蛋白作为诊断抗原，粪检血吸虫病阳性患者血清作为一抗，肝吸虫阳性患者血清作为考核交叉反应血清。结果 重组Calpain抗原免疫小鼠后，产生了一个极高的抗Calpain特异性抗体，免疫4周IgG类抗体达到高峰，与对照组鼠相比较免疫鼠血清中IgG1，IgG2a，IgG2b特异性抗体也显著上升；Calpain重组抗原血清检测日本血吸虫和粪检的符合率为100％，粪检阳性EPG低的个体抗Calpain抗体滴度高，EPG高的患者抗Calpain抗体滴度反而低，与肝吸虫的交叉反应率为37％。结论 日本血吸虫Calpain是一个具有免疫原性的蛋白，能够激发宿主产生高水平的免疫球蛋白，能够敏感地测定日本血吸虫的感染，提示Calpain可以发展成为日本血吸虫病的诊断抗原和日本血吸虫疫苗候选分子。     

日本血吸虫感染小鼠CD8+T细胞亚群的变化及其与肝脏免疫病理的相关性研究

目的:研究CD8+T细胞中Tc1(CD3+CD8+ IFN-γ+)?Tc2(CD3+CD8+IL-4+)和调节性CD8+T细胞(CD8+CD28-)亚群在日本血吸虫感染导致的小鼠肝脏免疫病理发生发展中可能的作用?方法:取正常小鼠及日本血吸虫感染8周的小鼠脾脏淋巴细胞,采用流式细胞术检测Tc1?Tc2和调节性CD8+T细胞亚群分别占T淋巴细胞的比例及绝对数量,并分析感染血吸虫不同时期的CD8+T细胞亚群与血吸虫感染小鼠肝脏肉芽肿和肝纤维化指标的关系?结果:与正常对照小鼠比较,日本血吸虫感染后小鼠脾脏的单个核细胞绝对数显著增加(P < 0.001);日本血吸虫感染小鼠脾细胞中T细胞(CD3+)和CD8+T细胞(CD3+CD8+)的比例及绝对数均显著增加(P均 < 0.05);此外,感染小鼠脾淋巴细胞中Tc1?Tc2?调节性CD8+T比例均显著增加(P均 < 0.001);与正常小鼠相比,感染小鼠Tc2/Tc1比率显著增加(P < 0.001)?此外,多元线性逐步回归分析显示Tc2和调节性CD8+T细胞是影响血吸虫感染所致肝脏免疫病理的主要影响因素?结论:CD8+T细胞及其Tc1?Tc2?调节性CD8+T细胞亚群在日本血吸虫感染宿主的肝脏肉芽肿和纤维化发生发展中可能发挥重要作用?其中,Tc2和调节性CD8+T细胞可能分别促进/抑制血吸虫感染所致的肝脏免疫病理?     

TORCH感染与自然流产相关性分析

目的　探讨TORCH系列病原体感染与自然流产的关系。方法　对 94例有自然流产史的和 50例无自然流产史的妇女 ,应用ELISA法检测血清中特异性TORCH系列IgG和IgM。 结果　观察组 94例中TORCH阳性 77例 ,检出率 81 .91 % ,而对照组仅8例阳性 ,检出率 1 6 % ,两组间差异有显著性。IgG阳性率分别是 :TOX 1 2 .8% ,RUV 2 0 .2 % ,CMV 39.4% ,HSV -Ⅱ 1 4 .9%。IgM阳性率分别是 :TOX 1 8.1 % ,RUV 2 4 .5 % ,CMV 41 .5 % ,HSV -Ⅱ 1 8.1 %。感染两种或两种以上TORCH病原体的 31例 ,占感染总数的40 .2 6 %。结论　TORCH感染与自然流产密切相关 ,对胎儿的危害程度与母亲的感染状况和孕龄有关 ,因此有必要对孕妇进行TORCH系列病原体的检测     

血清酸溶性蛋白定量在日本血吸虫病研究中的应用

本文研究动物感染日本血吸虫及毗喹酮治疗过程中血清酸溶性蛋白(ASP)浓度的动态变化。结果显示:家兔感染早期,血清ASP浓度明显升高,中期降至正常水平,末期再次明显升高,感染小鼠经吡喹酮治疗,血清ASP降低程腰与疗效、减虫率、病变消退等相符合。本文提示:血清ASP浓度测定可作为监护病情、判定药物疗效的指标,有一定的临床实用价值。     

Detection of a circulating SJ 70 antigen in human schistosomiasis japonica using a monoclonal antibody

A double antibody enzyme-linked immunosorbent assay (sandwich ELISA) was used for detection of a circulating antigen from human Schistosoma japonicum infections. This assay involves the use of poly clonal rabbit anti-schistosoma japonicum soluble egg antigen (SEA) antiserum to bind circulating antigen and a monoclonal antibody (MCAb-H4) to identify and quantify this antigen. Sera from 108 S. japonicum infected patients (acute and chronic) were tested. Sera from 93 of 95 (98%) patients with chronic infection were positive for this antigen; sera from 12 of 13 patients (93%) with acute infections were also positive. Antigen was not detectable in control human sera. Sera from 35 chronic schistosomiasis patients were collected between 6 and 12 months after paraziquantel treatment. Circulating antigen was not detectable in the sera from 33 of these patients (94%) and was dramatically reduced in the other two patients.