Application of SHSP60 for preparing diagnostic reagent for monitoring comprehensive hepatic pathology injury degree of schistosomiasis patient
A technology of damage degree and schistosomiasis, applied in the field of immunology, to achieve the effect of broad application prospects
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Embodiment 1
[0036] Example 1: In order to study whether the serum SjHSP60-IgG and its subtype IgG1 antibody titers are positively correlated with the average area of single egg granuloma and fibrosis degree of the corresponding liver in the rabbit model infected with Schistosoma japonicum.
[0037] 1. Put the snails in clean dechlorinated water at 25°C under light conditions to escape the cercariae. The cercariae were picked with an inoculation loop, moved to a cover glass (12mm×12mm), and counted under a dissecting microscope. Each group of mice was infected with 12, 20, 40 cercariae, respectively. The mice and rabbits were bound and fixed, and their abdomens were shaved. The skin was moistened with dechlorinated water, and the coverslip with cercariae was covered on it, and kept for 20 minutes. Mice and rabbits were sacrificed at 8 and 23 weeks after infection, respectively (Xu, Wen et al. 2010).
[0038] 2. Take the right anterior lobe liver. After being fixed with 10% neutral for...
Embodiment 2
[0041] Example 2: In order to study whether the serum SjHSP60-IgG antibody titer is positively correlated with the average area of single egg granuloma and the degree of fibrosis of the corresponding liver in the rabbit model infected with Schistosoma japonicum.
[0042] 1. According to the method in Example 1, each group of rabbits was infected with 20, 40 and 800 cercariae respectively. The level of SjHSP60 IgG antibody in serum was detected by ELISA.
[0043] 2. Then take the right anterior lobe liver. After being fixed with 10% neutral formaldehyde, routinely dehydrated and embedded in paraffin, the sections were stained with H.E. The granuloma area of a single egg was observed under a Zeiss microscope, and 30 areas were observed for each egg, and the average value was calculated. To evaluate the degree of liver fibrosis in infected rabbits, collagen fibers I, III and α-SMA were sectioned in paraffin. Then, an experienced pathologist evaluated the protein expression ...
experiment example 3
[0047] Experimental Example 3: In order to study whether the serum SjHSP60 antibody titer is also positively correlated with the degree of liver fibrosis in the schistosome infection mouse model and other factors that promote liver fibrosis.
[0048] 1. According to the method in Example 1 (1), the mice were infected with Schistosoma japonicum, and the mice were treated with carbon tetrachloride and PBS in different groups at the same time, and the levels of SjHSP60 IgG and its subtype IgG1 antibodies in the serum of the mice in each group were detected.
[0049] 2. Then take the right anterior lobe liver. After being fixed with 10% neutral formaldehyde, routinely dehydrated and embedded in paraffin, the sections were stained with H.E. The granuloma area of a single egg was observed under a Zeiss microscope, and 30 areas were observed for each egg, and the average value was calculated.
[0050] 3. SjHSP60 and MouseHSP60 were used to coat the plate with OVA protein, and ELIS...
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