Screen Serum Biomarkers With Relation To The HBsAg Carrier And Cirrhosis Research

Viral hepatitis is the strongest infectious disease ,it can result in Cirrhosis of liver. cirrhosis (Full name: Cirrhosis after hepatitis B virus infection ) is the sixth canse of ten causes of diseasing death in china . it is frequent-occurring and common diseasesAbove of them are none of special symptoms in the early stage .Once the patient with the typical symptoms and signs .Pathogenetic conditions tend to present at a late clinical stage with poor prognosis and therapeutic effects are not fine .For this reason, improving the level of early stage diagnosis and assessing the level of serious of them are reality meaning in clinical medicine.With the completion of the human genome sequence project and the highly speed development of molecule biology, Science research experienced tremendous transformation from genome to proteome .The development of knowledge and technology in proteomics has provided a new way to diagnosis of the disease in the early stage .Surface enhanced laser desorption -ionization time of fight mass Spectrometry (SELDI-TOF-MS) is one of the recently novel developed technologies,which to be able to analyse the complex biological sample powerfully .Render a strong and useful platform of research the occurrence and development of disease in proteomic .It has been already used in the early measuring and diagnosticing study of many kinds of diseases. To identify proteins in the Cirrhosis after hepatitis B virus infection (CHBVI) and HBsAg asymptomatic carrier (ASC)proteome that could potentially be used for detection of them and establish a serum protein fingerprinting technique coupled with a pattern-matching algorithm to identify CHBVI and ASC ,To provide new ideas of discovering disease markers of CHBVI and ASC ,To make diagnosis and the sourse and theory for the later research .Here we detect proteomes in serum from CHBVI, ASC and normal controls by SELDI-TOF-MS ,and try to use software to identify proteomic difference among CHBVI , ASC and the controls. While decision tree classification algorithms of CHBVI , ASC were determined by biomarker patterns software.Part one: Screening suitable protein chips for this research.Purpose: this part of research was to screen a suitable protein chip from two kinds of chips(CM10 and IMAC3).Method: Serum samples from 13 cases of CHBVI patients,13 cases of ASC patients and 13 cases of normal control people were collected respectively ,special protient or peptide spectra was determined by SELDI-TOF-MS measurement after treating the sample onto two kinds of protein chips for each case. According to the protein profiling results ,we screen suitable protein chips.Result : analysis of serum protein fingerprints show CM10 chip combined the most proteins, more than IMAC3 chip.Conclusion: CM10 chips can detect more significant difference protein peaks compared to IMAC3 chips and preferably apply to the research of the serum protein fingerprints.Part two:Screen serum biomarkers of CHBVI and ASC, Establish the diagnostic patternsPurpose:Discover the proteins with relation to CHBVI and ASC ,and provide new idea of finding new biomarkers of CHBVI and ASC, while establish diagnostic models of CHBVI and ASC, investigate its feasibility of the clinical application.Method:Serum samples were collected from 26 cases of the CHBVI ,28 cases of the ASC and 26 cases of normal people .the samples were profiled by SELDI-TOF-MS measurement using CM10 chips of ciphergen corporation .ciphergen biosystems software was used to analyze the proteomic profiles.Results:1.Comparison of the group of the CHBVI and the group of normal people : analysis two groups of 52 samples and get 69 proteins peaks, among them 25 protein peaks show the difference obviously in two groups compared with the normal people group ,9 proteins were up-regulated expression and 16 proteins were down-regulated expression in the CHBVI .Make use of those differenced proteins, we can get a model sensitivity of this model is 73%(19/26), specificity is 73%(19/26). Use this model to measure the 52 unknown serum samples. Put the serum protein mass sepctometry of each example into the model to estimate the result ,accurate rate is 73%(38/52).2.Comparison of the group of the CHBVI and the ASC: analysis two groups of 54 samples and get 69 proteins peaks , among them 26 protein peaks show the difference obviously in two groups . Compared with the ASC ,16 proteins were up-regulated expression and 10 proteins were down-regulated expression in the CHBVI. Make use of those differenced proteins, we can get a model sensitivity of this model is 92%(24/26), specificity is 86%(24/28). Use this model to measure the 54 unknown serum samples. Put the serum protein mass sepctometry of each example into the model to estimate the result: accurate rate is 89%(48/54).3.Comparison of the group of the ASC and the group of normal people: analysis two groups of 54 samples and get 69 proteins peaks , among them 28 protein peaks show the difference obviously in two groups .Compared with the normal people group ,5 proteins were up-regulated expression and 23 proteins were down-regulated expression in the ASC. Make use of those differenced proteins, we can get a model sensitivity of this model is 93%(26/28), specificity is 77%(20/26). Use this model to measure the 54 unknown serum samples, put the serum proteins mass sepctometry of each example into the model to estimate the result: accurate rate is 85%(46/54).4.Comparison of the group of the CHBVI ,the group of the ASC and the group of normal people ,we find two proteins that show the difference obviously in three groups .The m/z value of two proteins are 9422.89Da and 4656.59Da respectively. The protein the m/z value is 9422.89Da is up-regulated expression in the group of normal people, followed by the group of the ASC, and is down-regulated expression in the group of the CHBVI, The protein the m/z value is 4656.59 Da is up-regulated expression in the group of normal people, followed by the group of the ASC and is down-regulated expression in the group of the CHBVI.Those proteins the m/z value are 7774.40Da ,8150.87Da, 4604.42Da,9199.05Da, 4098.00Da are up-regulated expression in the group of normal people ,and down-regulated expression in the group of the ASC and the CHBVI. The protein the m/z value is 3478.03Da is up-regulated expression in the group of the ASC and the group of the CHBVI ,and is down-regulated expression in the group of normal people ,while the expression of this protein is no the difference obviously in group of the CHBVI and the group of the ASC.Conclusion:1.There is significant difference among the group of the CHBVI, the group of the ASC and the group of normal people in serum proteome, and those different proteins maybe the proteins related to the CHBVI and the ASC.2.Three models which established in this research can classify the patients of the CHBVI and normal people , the ASC and normal people , the ASC and the CHBVI, and screen the patients of the CHBVI from the patients of the ASC.3.The technology of SELDI-TOF-MS and protein chip combined with bioinformatics software show great potential for the detection of the ASC and the CHBVI in the clinical medicine .