华支睾吸虫病是一种危害严重的鱼源性人兽共患寄生虫病，全球约有3500万人感染，仅我国的感染人数就高达1500万。人体长期或慢性感染不但会引起肝脏组织的病变，更引发胆管上皮细胞的癌变，故国际癌症研究署(IARC)已将华支睾吸虫列为1类致癌物。目前对华支睾吸虫的致癌机制尚未明确，其在寄生过程中产生的排泄/分泌代谢产物被认为是重要分子，但这些代谢产物的成份十分复杂，有效分离其中致癌相关的蛋白和基因成为研究其致癌机制的关键。本研究除利用经典的噬菌体展示技术进行互作蛋白的筛选外，还结合了华支睾吸虫转录组EST数据分析以及双向电泳和质谱分离鉴定技术对华支睾吸虫排泄/分泌产物中的致癌相关蛋白进行筛选，并通过动物实验和细胞实验进行致癌特性鉴定，探索其在分子水平上的致癌机制。该研究不仅对胆管癌的病因学研究及预防具有重要意义，更可望为华支睾吸虫病的有效防控奠定理论与实验基础。

Clonorchiasis is a serious fish-borne zoonosis. Globally, nearly 35 million people were infected with Clonorchis sinensis, with approximately 15 million in China. Chronic infection can not only cause histopathological changes in liver, but also induce canceration of biliary epithelial cells, so C. sinensis has been categorized by the International Agency for Research on Cancer (IARC) as Group 1 carcinogens. The carcinogenic mechanism of this parasite is not particularly clear at present. The excretory-secretory(ES) products are considered as the most important factor, however, the composition of these ES products is complex, thus separation of ES products is the key to expound the carcinogenic mechanisms. In this study, we use classical phage display technology to screen the interacting proteins, and separate ES products by using 2-D electrophoresis and mass spectrometry identification technology combining with analysis of EST data. Characteristics of these proteins are identified by animal experiments and cell experiments. Above all molecular mechanisms of carcinogenesis can not only make a contribution to the etiology and prevention of cholangiocarcinoma, but also lay a foundation for the control and prevention of clonorchiasis.

华支睾吸虫病是一种危害严重的食源性人兽共患寄生虫病，人体长期或慢性感染不但可导致肝脏组织的病变，更可引发胆管上皮细胞的癌变，故国际癌症研究署(IARC)将华支睾吸虫列为1类致癌物。WHO食源性吸虫疾病全球负担报告称，据推算全球约有1531万华支睾吸虫感染者，每年约造成5500余例死亡。本研究拟在之前研究基础上进一步采用噬菌体cDNA表达文库技术和展示技术对华支睾吸虫童虫的标识性和保护性抗原基因进行大规模无渗漏筛选，从中鉴定出高特异性诊断抗原基因和强保护性抗原基因，为建立适用于人华支睾吸虫病临床免疫学检验及有效疫苗的研制奠定基础。本项目调整为应急管理项目，因此首先成功构建了华支睾吸虫童虫的cDNA 表达文库和T7 噬菌体展示cDNA 文库，为进一步华支睾吸虫童虫特异抗原的筛选和与胆管上皮细胞互作蛋白的筛选奠定了前提和基础。