Sensitivity of different isolates of Schistosoma japonicum from China to praziquantel.

Groups of C57BL inbred mice infected with each of the 4 different isolates, (Anhui, Hubei, Sichuan and Yunnan) of Schistosoma japonicum from the mainland of China were treated with praziquantel (PZQ) and the parasiticidal effects were compared. Worm reduction rate was recorded to assess systematically the sensitivity of 4 different isolates to PZQ in the mouse. Three dosage-levels of PZQ, ie 150, 230 and 310 mg/kg body weight in single doses were used. The worm development rates of control groups infected with schistosomes from Anhui, Hubei, Sichuan and Yunnan were 75.5, 81.8, 81.5, and 83.0%, respectively. At the dosage-level of 150 mg/kg, the worm reduction rates for the 4 different isolates were 36.0, 33.9, 25.5 and 35.6%, respectively. At the dosage-level of 230 mg/kg, the rates were 47.1, 46.0, 38.1 and 47.7%, while at the dosage-level of 310 mg/kg, they were 59.3, 58.6, 50.8 and 61.7%, respectively. The results indicated that the worm reduction rate of the Sichuan isolate was lower than that of the other three isolates, however, the differences were not statistically significant, suggesting that schistosomes of Anhui, Hubei, Sichuan and Yunnan isolates bear resemblance in drug response.     

中国大陆日本血吸虫地理株间成虫蛋白质组分的差异

目的分离、鉴定中国大陆日本血吸虫不同地理株间成虫差异表达蛋白。方法分别收集、制备日本血吸虫不同地理株成虫可溶性蛋白,经固相pH梯度双向凝胶电泳分离,凝胶银染,并用PDQuest8.0凝胶图像分析软件进行比较分析,筛选出差异表达蛋白点并采用基质辅助激光解析离子飞行时间质谱仪进行鉴定。结果湖南株、江西株和江苏株雄虫分别检出698±10、650±19、629±23个蛋白点,雌虫分别检出670±12、682±22、625±28个蛋白点,约90%蛋白点分子量处于20～90kD范围内,蛋白等电点在5～8之间。不同地理株雌虫与雄虫分别两两比较,雄虫发现14个差异点,雌虫发现18个差异点。对其中7个雄虫差异表达蛋白及3个雌虫差异表达蛋白经MALDI-TOF-MS鉴定分析,获得其肽质量指纹图谱、等电点、分子量等相关信息。7个雄虫差异表达蛋白分别为:SJCHGC00821蛋白、SJCHGC00475蛋白、3-磷酸甘油醛脱氢酶、谷胱甘肽转移酶片段、谷胱甘肽-S-转移酶、D-核酮糖-5-磷酸-3-表异构酶、G蛋白α亚基AgGq1,3个雌虫表达蛋白分别为:预测蛋白、GAF型传感器信号转导组氨酸激酶、组织蛋白酶B样半胱氨酸蛋白酶前体。结论中国大陆日本血吸虫不同地理株虫体间蛋白质存在差异,部分蛋白表达差异与虫体对吡喹酮敏感性密切相关。     

Compatibility between Oncomelania hupensis and different isolates of Schistosoma japonicum in China

Oncomelania hupensis from six localities were used for infection with different isolates of Schistosoma japonicum in the mainland of China, ie Anhui in the east, Hubei in the center, Guangxi in the south, Sichuan in the West, Yunnan in the southwest and Fujian in the southeast. Snails from Anhui and Hubei were readily infected with the local isolate of S. japonicum and cross infection also took place readily between the snails and the schistosomes from these two places. Snails from Sichuan and Yunnan were refractory to infection with schistosome isolates from Hubei and Anhui, but the isolates from Sichuan and Yunnan were able to develop in snails from Hubei and Anhui. Though the Guangxi isolate developed readily in both Anhui and Guangxi snails, the average precercarial period in the former was significantly longer than in the latter. None of the other snails from Sichuan, Yunnan and Fujian became infected. On the other hand, snails from Guangxi infected with Anhui parasites also had a longer precercarial period than that in Anhui snails. Snails from Fujian were readily infected with the isolates from Anhui and Yunnan. The present results suggest that there might be different geographic strains of S. japonicum and their Oncomelania snail hosts in the mainland of China.     

钉螺对不同宿主源性日本血吸虫毛蚴的易感性

目的了解钉螺对不同终宿主源性日本血吸虫毛蚴的易感性差异。方法收集家兔、水牛和小白鼠排出的虫卵,孵出毛蚴,以毛蚴和钉螺5：1的比例感染同一地钉螺。同时设对照组,观察各组钉螺感染率、死亡率。结果实验组家兔、水牛和小白鼠排出虫卵孵出毛蚴感染钉螺,获得的阳性率分别为1.42%、8.67%和19.87%,钉螺死亡率分别为29.5%、13.5%和24.5%;对照组钉螺阳性率分别为2.63%、2.02%和11.66%,死亡率分别为24.0%、49.5%和18.5%。结论不同宿主源日本血吸虫毛蚴对钉螺易感性存在差异。     

日本血吸虫感染不同相容性动物宿主的比较研究

目的 了解日本血吸虫感染不同动物宿主（包括天然宿主和实验动物宿主）后的虫体发育和宿主组织病理变化差异。方法 以黄牛、水牛、山羊、BALB/c小鼠、Wistar大鼠和新西兰大白兔为对象,同一时间人工感染相同来源的日本血吸虫尾蚴,收集感染后相同日龄的虫体（49d）,对虫体发育进行比较,对宿主的肝脏病变进行比较,并对黄牛、水牛和山羊的肝脏进行了病理切片的观察和比较。 结果 不同动物宿主来源的虫体的长度、宽度和发育情况存在着差异,总体来说是适宜宿主体内的虫体长度明显长于非适宜宿主体内的虫体长度,雌虫长于雄虫（Wistar大鼠来源的虫体例外）。发育情况除了Wistar大鼠来源的虫体未发育成熟,其余动物来源虫体均发育成成虫。六种动物宿主的肝脏病变存在很大的差异,适宜宿主感染后肝脏布满虫卵结节,病变尤其严重;而非适宜宿主的肝脏只有少量虫卵结节（水牛）,或者几乎看不到虫卵结节（Wistar大鼠）。天然宿主肝脏病理观察发现,黄牛组和山羊组肝脏组织炎性细胞明显增多且聚集,虫卵周围有大量嗜酸性细胞浸润,多量炎性淋巴细胞聚集浸润,形成典型条纹状嗜伊红沉淀物;而水牛组的肝细胞无变性、无炎症细胞的聚集浸润, 小叶结构完整。 结论 日本血吸虫对黄牛、山羊、BALB/c小鼠和新西兰大白兔的易感性高于水牛和Wistar大鼠,对前四种动物的致病性也强于后两种,本文为进一步了解日本血吸虫对不同终末宿主的感染差异提供了一些参考数据。     

中国大陆日本血吸虫品系的研究Ⅳ生物学性状的模？..

In this paper, a computerized numerical systematics was used to analyse the 5 different isolates of Schistosoma japonicum in the mainland of China by means of fuzzy clustering method with 32 quantitative data of each isolate about the morphological and biological characters. A dendrogram of the 5 isolates of S. japonicum was constructed on the basis of similarity coefficients as shown. The phylogenetic relationships revealed by the present study show that the two isolates of Anhui and Hubei get together firstly in one group, then the isolates of both Yunnan and Guangxi gather in another group, while the Sichuan isolate is closely related to the group of Anhui-Hubei isolates, not to the Yunnan ones, although both Sichuan and Yunnan isolates were in the southwest China.     

有螺无病地区钉螺与日本血吸虫相容性的研究

目的 研究安徽省宁国市有螺无病地区钉螺与日本血吸虫的相容性,探索有螺无病地区形成的原因.方法 采集宁国市有螺无病地区钉螺600只,随机分为2组,每组300只.在实验室条件下以本省日本血吸虫毛蚴进行钉螺群体感染,两组钉螺与毛蚴比例分别为1:20和1:40.以相同方法感染铜陵县的肋壳钉螺和泾县的光壳钉螺作为对照.感染后将钉螺置于室内22～24℃温度下饲养90d.观察各组钉螺感染率和死亡情况.结果 在1:20组和1:40组中,宁国市有螺无病地区钉螺均未能被日本血吸虫成功感染,而铜陵县的肋壳钉螺和泾县的光壳钉螺均感染成功.至观察期结束时,无论是自然死亡率还是感染后死亡率,铜陵县的肋壳钉螺均显著低于宁国市有螺无病地区钉螺和泾县的光壳钉螺(均P＜0.05),而后两地钉螺死亡率差异无显著性(P＞0.05).结论 当地钉螺与日本血吸虫不相容可能是宁国市有螺无病地区形成的主要原因之一.     

Dermal responses of various hosts to infection with Schistosoma japonicum cercariae

The present investigation concerns with the dermal reactions produced in various host species by the invading schistosomula in primary infection. Eight animal species including mouse, rat, hamster, jird, guinea pig, rabbit, rhesus monkey and pigeon were used. A small area of abdominal skin exposed to 300 Schistosoma japonicum cercariae was taken at different time intervals from each animal species.     

3种保虫宿主日本血吸虫特征性差异表达基因的筛选与验证

目的筛选日本血吸虫3种重要保虫宿主来源虫体的特征性差异表达基因,寻找可以用于鉴别这3种宿主来源虫体的遗传标记。方法日本血吸虫尾蚴人工感染黄牛、水牛和山羊,收集感染后49 d成虫,分别抽提RNA,逆转录成cD NA,体外转录为cRNA并进行片段化;采用定制的血吸虫芯片分别进行杂交,筛选每种宿主来源虫体的特征性差异表达基因。采用实时定量PCR法对所筛选差异表达基因的表达水平做进一步测定,验证芯片数据的结果,并对这些特征性差异表达基因进行验证。结果通过芯片杂交,分析筛选获得实验黄牛组来源虫体特征性差异表达基因3个,水牛组来源虫体4个,山羊组来源虫体7个。其中黄牛组2个、水牛组3个、山羊组5个特征性差异表达基因在另一批次3种宿主来源虫体的重复试验中得到验证。结论芯片杂交筛选出10个3种宿主来源虫体的特征性差异表达基因,这些基因可能可以作为这3种不同宿主来源虫体的遗传标记。     

日本血吸虫终宿主粪便中虫卵存活力的实验观察

目的 观察不同宿主粪便中日本血吸虫虫卵存活时间.方法 采集日本血吸虫虫卵阳性居民、水牛、猪、兔子和鼠等不同宿主粪便,观察不同宿主粪便中虫卵的孵化率,以及不同温度和不同时间内虫卵的存活情况.结果 来自猪、鼠、人、兔和水牛新鲜粪便中血吸虫虫卵的孵化率分别为78.9%、53.7%、41.7%、28.9%和22.1%.在37℃、30℃、20℃和4℃温度下不同时段粪便中虫卵存活率差异很大,其中干燥和样本粪便的大小是影响虫卵存活的重要因素.水牛和猪粪便中虫卵存活率高,随着温度的降低存活率逐渐增加.结论 不同宿主粪便中虫卵的孵化率,以及不同温度和不同时间内虫卵的存活情况存在显著性差异.     

Studies on the strain differences of Schistosoma japonicum in the mainland of China. II. Susceptibility of mammalian hosts

Six species of animals were percutaneously exposed to cercariae obtained from pools of naturally infected snails from different isolates of S. japonicum in the mainland of China as shown in Tables 1 and 2. Our results indicated that with the exception of rat, all animals under study were permissive host though their worm recovery rates varied with different isolates. The mean prepatent period in different host species were 35.0 +/- 0.8 to 36.4 +/- 1.0 days for Anhui isolate; 34.5 +/- 1.2 to 36.4 +/- 1.2 days for Hubei isolate; 34.5 +/- 1.3 to 35.8 +/- 0.6 days for Sichuan isolate; 35.1 +/- 1.0 to 37.3 +/- 1.9 days for Guangxi isolate and 36.1 +/- 1.9 to 37.8 +/- 0.8 days for Yunnan isolate. In general, the prepatent period was longer in the C57 BL inbred mice, hamsters and rhesus monkeys infected with Yunnan and Guangxi isolates, than that with Sichuan isolate. This result also indicates that the prepatent period of the strain of S. japonicum defined by Dr. Vogel as a Chinese strain which had been originated from the cercariae in infected Oncomelania hupensis hupensis snails from Jiaxing, Zhejiang (Kashing, Chekiang), China, and established in Tropeninstitut in Hamburg since 1937 by repeated passages in dogs and laboratory-bred O. h. hupensis snails, was 5-8 days longer than that of all the isolates under our study, probably due to some behavioral change. We suggest, therefore, that the Vogel's stock of S. japonicum should be termed as "Chinese Vogel strain".     

Susceptibility of Schistosoma japonicum to praziquantel in China

To look for possible evidence of the development of resistance in Schistosoma japonicum to praziquantel, we conducted a field study in China. During the non-transmission period of schistosomiasis a random sample of 2860 individuals from six villages in three provinces of China were examined using a parasitological stool examination. Of the 372 stool-positive subjects, 363 subjects were treated with a single oral dose of 40 mg/kg of praziquantel. Six to Seven weeks after treatment, of 334 subjects examined using the same stool examination, stool-negative results were found in 319 patients which represents a 95.5% parasitologic cure rate. Fifteen subjects still excreting eggs were treated a second time with the same dose of praziquantel. All stool samples, including those from participants re-treated with praziquantel, were re-examined 12 weeks after the first treatment and no stool-positive subjects were found. The results indicate that there was no evidence for reduced susceptibility of S. japonicum to praziquantel despite its extensive use in the main endemic areas of China for more than 10 years. The in vitro responses to praziquantel of cercariae, miracidia and eggs of S. japonicum compared with S. mansoni demonstrate that the cercariae, miracidia and eggs of S. japonicum are more sensitive to praziquantel than those of S. mansoni. More sensitive worms would be less likely to develop resistance and this could explain why no evidence for resistance was found in S. japonicum in China.     

湖南省日本血吸虫线粒体atp6基因部分序列的多态性分析

目的研究湖南省日本血吸虫不同地域自然隔离群线粒体ATP合成酶F0亚单位6基因（atp6）部分序列的遗传多态性,为湖南不同地域自然隔离群日本血吸虫的遗传特性研究提供实验依据。方法采用试剂盒抽提基因组总DNA,以特异性引物对线粒体atp6基因进行PCR扩增,通过单链构象多态性技术（SSCP）筛选出差异带型并进行测序,用DNAStar 5.0及Mega 4.0软件进行比对分析。结果湖南省5个流行区的日本血吸虫PCR扩增后获得了483 bpatp6部分序列,检测出17个变异位点,变异率为3.52%,雌雄虫之间的差异为0.0%～1.3%,不同地理来源虫株间的差异率为0.0%～1.5%。聚类分析结果表明：湖南不同地域自然隔离群日本血吸虫的种系发育不能按照其来源归为相应的系统发育进化树。结论湖南不同地域自然隔离群日本血吸虫的atp6基因存在明显的个体差异,是否由于生存环境的选择压力造成了同一地理来源日本血吸虫个体之间atp6基因的遗传差异,其原因尚有待于进一步研究。     

日本血吸虫感染适宜与非适宜宿主的免疫学特征初步研究

目的研究3种不同易感性宿主感染日本血吸虫后免疫应答特征的差异,初步探讨适宜和非适宜鼠类宿主感染血吸虫后免疫应答的机制。方法 C57BL/6小鼠、Sprague Dawley(SD)大鼠和东方田鼠(Microtus fortis)各12只,均随机分为感染组和未感染组,每组6只。C57BL/6小鼠、SD大鼠和东方田鼠的感染组每鼠经腹部皮肤分别感染日本血吸虫尾蚴20、200和1000条。感染后42d,剖杀各组动物,观察门脉系统成虫寄生及肝脏肉芽肿情况。收集血清,ELISA法检测细胞因子白细胞介素10(IL-10)、γ干扰素(IFN-γ)和血清特异性抗体IgG、IgG_(2a)及IgG_1的水平。结果感染日本血吸虫后42d,C57BL/6小鼠和SD大鼠均检获日本血吸虫成虫,并在宿主肝脏发现虫卵肉芽肿,而东方田鼠未检获血吸虫成虫及虫卵,肝脏正常。SD大鼠血清中IL-10的含量[(2.21±0.12)pg/ml]明显高于东方田鼠[(1.64±0.39)pg/ml](P<0.05)和C57BL/6小鼠[(0.10±0.04)pg/ml)](P<0.01),而东方田鼠也显著高于C57BL/6小鼠(P<0.01);SD大鼠血清...     

Ultrastructure of cultured cells from Schistosoma japonicum

Ultrastructures and their dynamic changes of the cultured cells from Schistosoma japonicum were observed in the present experiments. Several types-including polygonal, round granular, deltaic fan-shaped and flagellated cells-were found in the cultures. The polygonal cells took a major ratio in the cultures from adult S. japonicum, while the majority from schistosomula was round granular cells. The ultrastuctures on the cell surface were different between the cells from adults and schistosomula. Some papilla-like tubercula, microvilli and pinocytotic vesicles were observed on the surface of adult cells, but none were found on schistosomula cells. However, more or less mitochondria, endoplasmic reticula, ribosomes and glycogen were observed in the cytoplasm of the cultured cells from both adults and schistosomula. Golgi complexes were rarely found. The nucleus was round, with round nucleolus inside and clear pores on the unit membrane. There was much lumpish heterochromatin located near to the nuclear membrane. Cells from different worm tissues had their own organelles. The germ cells, vitelline cells, flame cells, multinucleate subtegumental cells and nerve cells could be observed in the cultures from adults. The vitelline cells were the greatest in number and nerve cells were the least in number among them. Similarly, there were germ cells, sustentacular cells, flame cells, nerve cells, mast cells, muscle cells, multinucleate subtegumental cells, interstitial cells and penetration gland cells in the cultures from the schistomomula. In addition, a few division cells were also found. It indicated that the schistosomula cells had greater potential ability to proliferate than the adult cells in in vitro culture. Along with the prolongation of the culture time, degeneration of schistosomal cell occurred more and more. Generally, the electron density of cultures gradually got lower, the cristae of mitochondria blurred and disappeared and the mitochondria themselves swelled and finally vacuoled completely. Vitelline cells were most sensitive to the changes of the in vitro condition in all cultures. Their degeneration showed the following characteristics: (1) vitelline globules fused each other, the space between vitelline globules and the membrane surrounding them broadened gradually and vitelline globules were released and uncovered; (2) rough-surfaced endoplasmic reticula enlarged, vacuolated and the ribosomes dropped; and (3) the number and volume of lipid increased. The ultrastructural changes of most of the cultures from schistosomula had the following trends: (1) heterochromatin increased and euchromatin decreased gradually; and (2) endoplasmic reticula changed into short tubes and vacuoles and disappeared finally. The degenerative process of the cultures from S. japonicum consisted of necrosis according to the ultrastructural changes of the mitochondria, vitelline globules, chromatin and endoplasmic reticula within the cells. The changes of the above structures could be used to estimate whether the culture conditions were appropriate.     

不同阶段日本血吸虫对吡喹酮敏感性的研究

目的探索成虫、尾蚴、毛蚴、虫卵等不同阶段日本血吸虫对吡喹酮的敏感性.方法于血吸虫病非流行季节,在江西、湖北、湖南6个村,采用吡喹酮40 mg/kg顿服治疗粪检阳性者363例,以观察日本血吸虫成虫对吡喹酮的敏感性.采集阳性粪便收集虫卵,孵化毛蚴,感染钉螺收集尾蚴.观察尾蚴在10-5、6×10-7、4×10-7、10-7mol/L4种浓度吡喹酮溶液中断尾率.毛蚴在5×10-6、10-6、5×10-7、10-7 mol/L 4种浓度吡喹酮溶液中的形态改变.以及虫卵在5×10-6、10-6、5×10-7、10-7mol/L 4种浓度吡喹酮溶液浸泡24 h后的孵出率.结果治疗6～7周后复查334例,319例阴性,阴转率95.5%,2次治疗阴转率达100%.尾蚴在10-5mol/L吡喹酮溶液中出现强烈挛缩、断尾,40 min断尾率为96.6%～100%.毛蚴在5×10-7mol/L吡喹酮溶液中发生形态改变,5 min形态改变率92.3%～100%.虫卵在5×10-6mol/L吡喹酮溶液中浸泡24 h后孵出率为0.结论与曼氏血吸虫相比较,日本血吸虫成虫、尾蚴、毛蚴和虫卵对吡喹酮更为敏感.     

不同血吸虫病流行区人群循环抗原和抗体检测结果分析

血清学诊断作为血吸虫病治疗、扩大化疗及疗效考核的依据已被采用。检测循环抗体 (CAb)和循环抗原 (CAg)因其敏感性和特异性好 ,操作简便、快速 ,结果准确而被认可[1 - 3] ,并用于血吸虫病诊断和血清流行病学调查。本文对不同血吸虫病流行区人群进行检测的结果报告如下。1　受检对象和检测方法1.1　受检对象血吸虫病流行区、基本控制区及非流行的对照区受检对象 ,分别选择荆州市、枝江市及宜城市的机关工作人员和农村常住居民。1.2　检测方法CAb和 CAg检测 ,分别用血吸虫病抗体快速测定试剂盒和单克隆抗体斑点酶联免疫吸附试验诊断血吸…