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Objective: proteomics detected in healthy volunteers (Ⅰ) with chronic hepatitis B (referred to as chronic hepatitis B) damp heat resistance (II), liver depression and spleen (III), blood stasis network (VI), liver and kidney Yin (V) and spleen deficiency (Ⅳ) five TCM peripheral blood of patients differentially expressed proteins in the pulp, to analyze the relationship between these differentially expressed proteins and liver function and inflammatory reaction condition, chronic initially explored hepatitis B TCM syndromes the inflammatory reaction conditions proteomics law for TCM Clinical Treatment of Chronic Hepatitis B available reference objective indicators, and ultimately provide a scientific basis to further investigate chronic hepatitis B TCM syndrome essentially. : Accordance with viral hepatitis the TCM Syndrome standard \blood stasis, liver and kidney and spleen and kidney yang Five TCM patients with a total of 100 cases of the observation group, five cases of healthy subjects as the control group. Using two-dimensional electrophoresis technology in the PH4-7 IPG gel DryStrip to about 130μg sample volume, electrophoresis conditions of 75000v.h preliminary better resolution plasma protein expression profiling. Gel imaging system of differential protein spots were scanning plasma protein expression profiling analysis ImageMaster2D software. Experimental group and the normal control group standardized total gray value (% Vol) difference of up to 2 times or more, and chronic hepatitis B five kinds of traditional Chinese medicine syndrome the apparent regularity differentially expressed protein spots as differentially expressed proteins, by matrix assisted laser desorption time of flight mass spectrometry (MADLDI-TOF-MS) to detect the differentially expressed proteins, binding of each protein points relative to the isoelectric point (PI), the relative molecular weight (Mw) to Swis s-prot protein database match retrieval, identified the differentially expressed protein. Screening TCM syndromes in chronic hepatitis B five kinds the obvious regularity differentially expressed and embody the condition of the liver inflammatory response protein by ELISA method to expand the sample size syndrome protein expression to validate the above differences. Measurement data with X ± S) SNK-q statistical methods to analyze all the data are in the computer processing SPSS v15.0 statistical software. Results: (1) to obtain the resolution and reproducibility was good plasma proteome two-dimensional electrophoresis silver staining. (2) through the comparative analysis of the normal control group, damp heat resistance syndrome group of patients with chronic hepatitis B, stagnation and spleen deficiency syndrome group, blood stasis syndrome group, the liver and kidney syndrome group and spleen deficiency syndrome group dimensional electrophoresis profiles found that there are a the apparent regularity differentially expressed protein spots were a total of five, five syndromes were identified by mass spectrometry haptoglobin protein (HP), immunoglobulin J chain (Ig-J), retinol binding protein ( RBP), C-reactive protein (CRP), vitronectin protein (VN). Its function is mainly related to the decline in hepatic synthetic function, inflammation, immune pathological damage. (3) screening in the hepatitis B five kinds of traditional Chinese medicine syndrome there are the obvious regularity differences in expression and liver inflammation status embodies protein CRP, RBP and VN (CRP and liver inflammation-related, the RBP and VN degree of liver damage that liver closely related functions), the above differences in protein expression by ELISA method to verify expand the sample size syndrome, syndromes group retinol binding protein (RBP) relatively normal control group dropped its reducing the order of ( reduce the maximum row before) Ⅵ gt; Ⅴ gt; Ⅳ gt; Ⅲ gt; Ⅱ. TCM syndrome waiting group and normal control group were significant difference (P lt; 0.05) TCM designate group comparison Ⅳ and Ⅲ between there is a significant difference (P lt; 0.05); glass even protein (VN) relative the normal group declined, reducing the order (reduce the maximum row before) VI gt; Ⅴ gt; the gt; Ⅳ II gt; III. TCM syndrome waiting group and normal control group were significant difference (P lt; 0.05) TCM designate group comparison Ⅵ and Ⅴ between there are significant with difference (P lt; 0.05), Ⅱ and Ⅲ there are significant differences (P lt; 0.05); relatively normal control group were elevated C-reactive protein (CRP), higher order (higher up, the top) Ⅵ gt; the Ⅴ gt; Ⅳ Ⅲ gt; Ⅱ. Each TCM syndrome group and normal control group, there were significant differences (P lt; 0.05), comparison between TCM syndrome group I and V were significant differences (P lt; 0.05). Conclusion: (1) to obtain the resolution and reproducibility was good plasma proteome two-dimensional electrophoresis silver staining, the initial establishment of chronic hepatitis B stagnation and spleen deficiency group, damp heat resistance group, the group of liver-kidney, spleen and kidney yang group, blood stasis group and the healthy control group plasma proteome expression profiling. (2) there are several differences in protein expression between chronic hepatitis B five TCM and normal, show that CHB existence of multi-protein exceptions, this is the essence of traditional Chinese medicine, \(3) by ELISA method to expand the sample size syndrome cases the expression above differences reflect the extent of the damage of liver inflammation and liver function in protein CRP, RBP and VN verify discovered protein which reflect the body's inflammatory response status - CRP group of relatively normal elevated, elevated to the order of the (higher up, the top) Ⅵ gt; the Ⅴ gt; Ⅳ gt; Ⅲ gt; Ⅱ prompted CRP in differential expression of the inflammatory response with chronic hepatitis B five TCM climate conditions there is a certain regularity, that stasis stasis inflammatory response syndrome group heaviest, followed by the liver and kidney syndrome group, spleen deficiency syndrome group, stagnation and spleen deficiency syndrome group, liver and gallbladder damp-heat syndrome group relative to the lightest. Reflect the damage of liver function the protein - RBP and VN relatively normal group decreased, consistent lower order, respectively (reduce the maximum row before) VI gt; the Ⅴ gt; Ⅳ gt; Ⅲ gt; Ⅱ and Ⅵ gt; Ⅴ gt; Ⅳ gt; Ⅱ gt; Ⅲ. Are blood stasis obstructing collaterals lowest, followed by liver kidney yin deficiency, spleen deficiency syndrome, and liver depression and spleen deficiency syndrome, hot and humid the highest relative retention syndrome, suggesting that the RBP and VN differentially expressed in patients with chronic hepatitis B five TCM climate liver function between the extent of the damage there is a certain regularity, that blood stasis syndrome group liver function damaged the most important, followed by liver kidney yin deficiency, spleen deficiency syndrome, and liver spleen deficiency syndrome in the hot and humid retention syndrome, liver dysfunction is relatively light. (4) chronic hepatitis B five syndromes, CRP, RBP and VN three indicators relative to the normal control group increased and reduced order consistent (CRP ↑: Ⅵ gt; Ⅴ gt; Ⅳ gt; Ⅲ gt; Ⅱ ; RBP ↓: Ⅵ gt; Ⅴ gt; Ⅳ gt; Ⅲ gt; Ⅱ: VN ↓: Ⅵ gt; Ⅴ gt; Ⅳ gt; Ⅱ gt; Ⅰ Ⅱ), patients with liver inflammatory reaction prompted chronic hepatitis B five TCM climate heavier positive correlation between the degree of liver dysfunction, inflammatory response, the more severe the degree of impairment of liver function.
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