Proteomics Analysis And Sensitivity Of Praziquantel Of Different Isolates Of Schistosoma Japonicum In Mainland Of China

Schistosomiasis is a serious parasitic zoonosis caused by worms of the Genus Schistosoma,which can infect human being. China is schistosomiasis japonica epidemic area, the disease had been prevalent in Yangtze River and the south of 12 provinces of China. After nearly 60 years of efforts, the epidemic has been effectively controlled, through synthesize measures, such as to control and detect snails, to detect and treat disease, to improve drinking water and lavatories and even to creat better environment. But, at present, there are still seven provinces that have schistosomiasis prevalence in our country. Chemotherapy is an important method to reduce the damage of schistosomiasis patient's condition and control its dissemination. Especially, the widespread use of praziquantel, a high effective and low-toxic drug , played a key role in schistosomiasis control. However, on account of the increase of in highly epidemic areas to take a widely range of praziquantel, with the same susceptible population to increase the frequency of repeatedly taking praziquantel, as well as the increase in implementing nationwide chemotherapy to take inadequate doses of praziquantel , so monitoring on Schistosoma of sensitivity to praziquantel and the related research has also been highlighted. Investigation and mechanism research of different geographical isolates of Schistosoma japonicum to praziquantel sensitivity have great significance to monitor and prevent of resistance to praziquantel.This article taken from 3 geographical isolates of Schistosoma japonicum of mainland of Jiangsu, Jiangxi and Hunan Provinces adult worms carried out research work as follows:1 To campare the sensitivity of different isolates of Schistosoma japonicum to praziquantel1.1 Objective: To monitoring the sensitivity of different isolates of Schistosoma japonicum to praziquantel .1.2 Method :with 3 different isolates of Schistosoma japonicum cercariae infecting the mice, after 35d infections, to respectively give 31.25mg/kg×3d, 62.5mg/kg×3d, 125mg/kg×3d, 250mg/kg×3d and 500mg/kg×3d dose of praziquantel by different groups. After 15d of administration, to anatomy infected mice and calculate worm reduction rate of each groups, and through the different groups reduction rate of worms to calculate the praziquantel ED50 value of different isolates of Schistosoma.1.3 Results: Using graphpad prism 5 application software to fitting dose-response relationship of sigmoidal shape curve, and also calculate praziquantel median effective lethal (ED50) of the three isolates of schistosoma japonicum respectively : the isolates of Jiangsu S.j is 68.87mg/kg, and its 95% confidence interval is 60.51 to 78.38mg/kg; the isolates of Jiangxi S.j is 133.3mg/kg, and its 95% confidence interval is 112.5 to 157.8mg/kg; the isolates of Hunan S.j is 84.18mg/kg, and its 95% confidence interval is 73.98 to 95.78mg/kg.1.4 Conclusion: praziquantel sensitivity of different isolates of Schistosoma japonicum in mainland of China has obvious differences, and the sensitivity of praziquantel of the isolates of Jiangxi S.j is the worst.2 Proteomic analysis of different isolates of Schistosoma japonicum in mainland of China2.1 Soluble Proteins analysis of different isolates of Schistosoma japonicum in mainland of China by two-dimensional electrophoresis analysis2.1.1 Objective: To compare the adult proteomic of different isolates of Schistosoma japonicum in mainland of China2.1.2 Methods: use three differences isolates of Schistosoma japonicum ceraariae to infect rabbits ,and then collect the male and female worms and the soluble proteins were prepared from the different isolates of Schistosoma japonicum adult worms.After two-dimensional (2-D) gel electrophoresis was performed, the results were analyzed by PDQuest 8.0.2.1.3 Results: There were (698±10), (650±19) and (639±23) protein spots detected from three geographical isolates of male S.japonicum of Hunan, Jiangxi and Jiangsu respectively, and (670士12), (682±22) and (625±28) protein spots from female. About 90% of proteins had a molecular weight of 20KD to 90KD, and theoretical pI (isoelectric point) values of 5 to 8. The pairwise comparison between different isolates of the female and male worms showed that there were 14 differentially expressed protein spots in male worms, In Hunan and Jiangsu isolates there is 1 separate protein spots respectively, 1 protein spot did not express in Jiangsu isolates and the other 11 protein spots are upregulated or downregulated; and 18 spots in female worms, including 1 independent expressed spot in Jiangsu isolates and 1 protein spot did not express in Hunan isolates ,and the other 16 protein spots are upregulated or downregulated.2.1.4 Conclusion: There are different protein expresse among the different isolates of Schistosoma japonicum in mainland of China.2.2 Identification of the differentially expressed soluble protein in different geographical isolates of Schistosoma japonicum by MS.2.2.1 Objective: Identification and functional analysis of differentially expressed proteins.2.2.2 Methods: Cut and enzyme the differentially expressed protein spots from two-dimensional electrophoresis gel, and then use MS to identify the enzyme derived peptides, and combine with bioinformatics to analyse the function of the identified protein.2.2.3 Results: The selected 22 differentially expressed protein spots were identified by mass spectrometry analysis, there are 10 protein spots which were successfully determined peptide fingerprints, including 7 males and 3 females differentially expressed proteins. The results showed that seven male differentially expressed proteins identified were: SJCHGC00821 protein, SJCHGC00475 protein, glyceraldehyde 3-phosphate dehydrogenase, glutathione transferase, Glutathione S-Transferase, D-ribulose-5-phosphate 3-epimerase, G protein alpha subunit AgGq1, and 3 female proteins were: predicted protein, GAF sensor signal transduction histidine kinase and Cathepsin B-like cysteine proteinase precursor.2.2.4 Conclusion: The identified 10 different proteins are mainly related to the processes of protein post-translational modification, protein metabolism, carbohydrate metabolism, signal transduction, cell regulation and energy metabolism and so on. it is worth further study that some differentially expressed proteins are closely related to the sensitivity of praziquantel to worm.