Study On Form Mechanism Of Fibrosis Capsule Wall Around Hepatichydaid

Object:After E.G invaded into human body,it will form focus of infection-Cy.In the process,liver will have some reactions,which includes apoptosis of cells, immune response,and so on.there will be appearance of fibrous structure,the part near to E.G called capsula externa and the part near to liver called adventitia.We want to find reasons of fibrous structure and find some related genes or protein.1. we want to find some difference.between normal and disease liver proteome through contradistinction.2. Using software of bioinformation to analysis SSH cDNA sequence and copy cDNA Lib.Methods:1. Through 2-DE technology,we want to find difference between proteome.2. we hope use SSH to find some difference in mRNA.3. Copy cDNA Lib through the cultivation of bacteria.4. Use software of bioinformation to get information of gene sequence and protein.5. Trough hybridization of RNA spots,we check the different genes.Results:1. Because of specificity of disease liver,we improved the methods of extracting proteome and got one suitable method,which include components and CO of lysate and extracting methods.Now,we used the lysate of 8M Urea to dissolve proteome,use hypersound to clear nucleic acid and through 45000g centrifugal force coμLd ensure the integrity of proteome.2. we regulated capacity of sample,choose right pH of strips,find suitable conditions of IPQchoose better method of dyeing,and so on.we got one suitable method of 2-DE about disease liver.we used 800μg sample,strip of pH5-8,80000Vhr of IPG and R-250 to get some good atlases.3. We got 72 protein spots through PDQuest and MALDI-TOF-MS and check names and descriptions of 42 protein in NCBI database.these protein include eleven variety. Material and energy metabolism account 16%,development of focus of infection and immune response each account 10.5%, Apoptosis of cells or Inflammatory reaction and translocation of materials each account 7.9%, biosynthesis, drμg resistance, direction of liver injury, conduction of signal each account 5.3%,unknown protein accout 7.7%4. We referred 67 EST sequences to NCBI database from SSH cDNA Lib through analysis of Seqman,DNAstar and ESTin.Genebank ID:G0349048-G0349115.we also got different cDNA sequence of 20 genes.5. We copy the cDNA lib twice and reduced the probability of contamination.6. We checked expression of 5 genes in disease liver.P40,P150 and cytochrome c oxidase subunitⅢhave high expression level, ATP synthase F0 subunit 6 and cysteine protease have low expression level.Conclusion:1. We find some protein educe some effects in disease.such as protein of immune response, Apoptosis of cells or Inflammatory reaction,translocation of materials,and so on.their expression in disease liver are different.2. We checked expression of 5 genes in disease liver throμgh SSH and hybridization of RNA spots,we think they may have association of Inflammatory reaction.