1.Detection of major allergen parvalbumin in fish
Qiufeng CAI ; Guangming LIU ; Ruifang WANG ; Xichang WANG ; Wenjin SU ; Minjie CAO
Chinese Journal of Immunology 2000;0(08):-
Objective:To develop an indirect enzyme-linked immunosorbent assay (ELISA) for semi-quantification of major allergen parvalbumin in fish.Methods:The soluble proteins were prepared from both white and dark muscles of seven species of freshwater fish and five species of marine fish.Tricine-SDS-PAGE and Western blot were performed to examine the protein patterns of fish muscle extracts.Natural parvalbumin being used to make calibration curve was purified from silver carp (Hypophthalmichthy molitrix) by ammonium sulphate fractionation,followed by ion exchange and gel filtration chromatography.The molecular mass of purified protein was estimated by Tricine-SDS-PAGE and identified by Western blot with anti-frog parvalbumin monoclonal antibody PARV-19.ELISA using PARV-19 was carried out to evaluate parvalbumin contents in white and dark muscles.Results:Tricine-SDS-PAGE revealed species-specific differences in proteins of heated extracts.Western blot confirmed that the major bands were showed in Tricine-SDS-PAGE with the molecular masses of 10-14 kD corresponded to parvalbumins recognized by PARV-19 and various numbers of isoforms of parvalbumin existed in different species of fish.There might be some differences in the parvalbumin contents and the epitope region was recognized by PARV-19 based on the differences in relative intensities of protein immunodetection.The ELISA showed that the contents of parvalbumin were 4 to 33 folds higher in the white muscle than in the dark muscle and varied greatly in different species of fish.Conclusion:These results validate that the dark muscle might be less allergenic than the white muscle due to the lower content of parvalbumins,and it is suggested that the commercial anti-parvalbumin antibody PARV-19 can be used to detect parvalbumins from the commercially important species of fish tested in this study and the method we develope succeeds to detect the major allergen in various species of fish.
2. Association between the internal exposure levels of phthalates and executive function of preschool children
Xiuxiu CAI ; Hui GAO ; Shuangqin YAN ; Jie SHENG ; Chunli GU ; Zhongxiu JIN ; Juan QI ; Yuanyuan XU ; Qiufeng ZHANG ; Hui CAO ; Fangbiao TAO ; Jiahu HAO
Chinese Journal of Preventive Medicine 2018;52(3):296-302
Objective:
To investigate the association between the internal exposure levels of phthalic acid esters (PAEs) and executive function (EF) of preschool children.
Methods:
Between October 2008 and October 2010, pregnant women who accepted pregnancy health care services in four municipal medical and health institutions in Ma'anshan city, Anhui Province, were recruited as study objects. A total of 5 084 pregnant women and 4 669 singleton live births were enrolled in this cohort. The follow-up study was conducted from April 2014 to April 2015. A total of 3 725 data-completed preschool children aged 3 to 6 years older entered in this study. The method of analysis seven metabolites of phthalates in urine was high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and divided objects into low (