Objective To analyze and identify the proteins relevant to Ganyang Huafeng Syndrome(GYHF) and Yinxu Shengfeng Syndrome(YXSF) of cerebral infarction,and investigate the essence of GYHF and YXSF.Methods 3 groups,GYHF of cerebral infarction,YXSF of cerebral infarction and healthy people were designed with 15 cases of each group.Total protein were extracted from lymphocyte of peripheral venous blood,and two dimensional electrophoresis was used to separate total proteins from norma land model ratsynovium.Having been stained,proteins were analyzed by PDquest software.Some selected protein spots were identified by peptide mass fingerprint(PMF) based on Matrix assisted laser desorption/ ionizationtime of flight mass spectrometry(MALDI-TOF-MS) and database.Results 23 different protein spots and 8 identical peotein spots were found out from GYHF and YXSF group,22 proteins were identified from the different protein spots including 15 kinds of proteins and 6 protein spots including 5 kinds of proteins.GYHF of cerebral infarction related with proteins such as metabolic enzymes,signal transmission proteins,cytoskeletal protein,antioxydation proteins and so forth.Conclusion GYHF and YXSF of cerebral infarction have different proteins and identical proteins,which indicates that the two syndromes have the same matter substructure and different essential connotation.

BACKGROUND:Vascular endothelial growth factor is found the strongest to promote angiogenesis, which show strong expression in synovial cells in rheumatoid arthritis.
OBJECTIVE:To evaluate the relation between the expression of vascular endothelial growth factor and microvessel density in rheumatoid arthritis.
METHODS:Col agen II and complete Freund’s adjuvant were used to induce experimental arthritis in Sprague-Dawley rats. Synovium expression of vascular endothelium growth factor and microvessel density were assayed at different time points using immunohistochemistry method, and changes in new vessels, vascular endothelial growth factor expression and arthritis index were observed along the time course. The correlations between vascular endothelial growth factor expression and number of new blood vessels as wel as between arthritis index and vascular endothelial growth factor expression were studied with linear correlation analysis and rank correlation analysis, respectively.
RESULTS AND CONCLUSION:With the time lasting, new blood vessels in the synovium proliferated, the synovium was thickened, and the arthritis index rose gradual y as wel as vascular endothelial growth factor expression and number of new microvessels were on the rise. Vascular endothelial growth factor expression was positively correlated with arthritis index (r=0.535, P<0.05) and significantly positively correlated with number of new microvessels (r=0.860, P<0.01). Vascular endothelial growth factor is involved in the course of synovium pannus response, and vascular endothelial growth factor and microvessel density may play an important role in rheumatoid arthritis pathogenesis.

BACKGROUND: Pathological change of synovium in rheumatoid arthritis (RA) has the characteristic of tumor-like growth, it appears thickening of the synovium tissue and the formatiom of pannus, which generate periarticular erosion and destruction. Multiplicate cell factors and growth factors participate in the development course of tumor-like lesion of synovium, and the tumor necrosis factor-α(TNF-α) and vascular endothelial growth factor(VEGF)play important roles in the development of RA and the formation of pannus.OBJECTIVE: To observe the contents of plasma TNF-α of collagen-induced arthritis and the expression change of VEGF of synovium at different time point, and investigate the effect and correlation of TNF-α and VEGF in the pathogenesis of RA.DESIGN: Randomized grouping experiment taking animals as subjects.SETTING: Institute integrated traditional and western medicine of Xiangya Hospital of Central South University.MATERIALS: The experiment was finished from July to November 2003 in the laboratory of institute integrated traditional and western medicine.Forty SD rats aged 45-50 days were selected, the rats were randomly divided into the normal control group (n=10) and collagen-induced arthritis model group(n=30).METHODS: 10 mg Cattle collagen type Ⅱ and 5 ml full Freund adjuvant were grinded together, 100 μL of which were intradermally injected at the root of tails of rats, collagen-induced arthritis model of rats were re-immunized with the above-mentioned methòd and dosage after 21 days. The accumulated points of arthritis index was evaluated based on degree and extent of flare and the condition of tumefaction and deformation, the higher the accumulated points of arthritis index were, the more serious the arthritis symptom was. The blood was obtained by decapitation after 25 days in normal control group, and in the collagen-induced arthritis model group the blood was obtained by decapitation 25,30,35,40,45 days after immunization (model establishing), the contents of plasma TNF-α of collagen-induced arthritis rats at different time point was detected by radio-immune assay, the level of expression of VEGF in synovium tissue were detected with immunohistochemical method simultaneously, the correlation of invasion time and the neovascularization of synovium. The accumulated points of arthritis index . TNF-α and VEGF was observed. The correlation of TNF-α and VEGF was analyzed with the linear correlation analysis, the correlation of the accumulated points of arthritis index and TNF-α. VEGF was analyzed with the rank correlation analysis.MAIN OUTCOME MEASURES: The correlation of invasion time of collagen-induced arthritis with the accumulated points of arthritis index, with the plasma content of TNF-α and the expression of VEGF, the correlation analysis of the plasma content of TNF-α of collagen- induced arthritis rats with the expression of VEGF in synovium.RESULTS: Forty rats attended the experiment, all of them entered the final analysis. the neovascularization of synovium was increased, synovium was thickening, the accumulated points of arthritis index was gradually increased, and the contents of TNF-α and level of VEGF were increased accordingly with the process of the invasion time of the collagen-induced arthritis rats; Its accumulated points of arthritis index had the positive correlation with the level of expression of VEGF (r=0.535 ,P ＜ 0.05)and had the correlated increasing tendency with the contents of TNF-α, but there was no significant difference(r=0.371 ,P ＞ 0.05 ). the plasma contents of TNF-α had the positive correlation with the level of expressions of VEGF (r=0.893,P ＜ 0.01 ).CONCLUSION: The TNF-α and VEGF have an important effect on the inflammatory reaction of RA and Cytokine network of neovascularization of synovium, they are possibly mutually influenced and promoted and have the effect of mediating the malignant network circulation; They are the key factors among multiplicate ones which mediate the generation and development of RA, bone erosion and Mutilation.

Objective:With proteomic approach,to find the differentially expressed proteins spots in the peripheral blood mononuclear cells(PBMCs) of healthy people and intracerebral hemorrhage patients with liver yang forming wind syndrome(LYFWS) treated by Zhenganxifeng decoction(ZGXFD) to investigate the possible proteins of LYFWS and the cure mechanism of ZGXFD.Methods: 10 intracerebral hemorrhage patients with LYFWS were treated by ZGXFD for 7-10 days.10 healthy people were selected taken as healthy controls.10ml venous blood was drawed from every subjects and PBMCs were isolated from blood by using lymphocytes separation medium.The total proteins was extracted from PBMCs.The total proteins from either intracerebral hemorrhage patients or healthy controls were prepared by means of immobilized pH gradient based on two-dimensional gel electrophoresis.After Coomassie brilliant blue G250 staining,gel-image analysis was performed by PDQuest.The differentially expressed proteins spots were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry(MALD I-TOF-MS).Results: Gel-image analysis revealed that there were 22 proteins spots expressed differentially.20 proteins spots were expressed differentially in the intracerebral hemorrhage patients with LYFWS as compared with healthy control,of which 8 proteins were down-regulated,7 proteins were up-regulated and 5 proteins were lost.After treated by ZGXFD,4 proteins were down-regulated,5 proteins were up-regulated and 2 proteins were lost,9 proteins weren’t expressed di erentially as compared with healthy control.8 of 20 di erentially expressed proteins spots in the intracerebral hemorrhage patients with LYFWS identi ed by MALDI-TOF-MS as compared with healthy control.These proteins are related to cell metabolism,signal transduction and so forth,of which 6 proteins were down-regulated,1 proteins were up-regulated and 1 proteins were lost.After treated by ZGXFD,2 proteins were down-regulated,1 proteins were lost,5 proteins weren’t expressed di erentially as compared with healthy control.Conclusion: 2-DE pro les of intracerebral hemorrhage patients with LYFWS in PBMCs had been established.The proteins of intracerebral hemorrhage patients with LYFWS had been identi ed,which were related to cell metabolism,signal transduction and so forth.This research presented that ZGXFD could adjust multiple di erentially expressed proteins of intracerebral hemorrhage patients with LYFWS.It was also indicated that the cure mechanism of ZGXFD was to regulate multitarget proteins.

Objective To explore the related protein which lead to rheumatoid arthritis (RA) and to find different proteins associated with active RA by comparing the expression levels of proteins in the peripheral blood mononuclear cells (PBMCs) of healthy individuals to patients with rheumatoid arthritis using a proteomics approach. Methods Samples of peripheral blood were collected from 9 patients diagnosed as active RA and 9 healthy individuals. PBMCs were isolated from blood using lymphoeytes separation medium. The total protein was extracted from the peripheral blood mononuclear cells. The total protein from either RA patients or normal controls was prepared by means of immobilized pH gradient based on two-dimensional gel eleetrophoresis. After Coomassie brilliant blue G250 staining, gel-image analysis was performed by using PDQuest.The differentially expressed proteins were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALD I-TOF-MS). Then APOA-I was validated by RT-PCR. Results 2-DE patterns of PBMCs from controls and RA patients were presented. The results showed that the average number of protein spots was 556 and 579 respectively, and the corresponding average matching rate was 89.4％ and 88.5％ respectively. Gel-image analysis revealed that there were 23 differential protein spots. Fourteen of total 18 differential protein spots were successfully identified by MALD I-TOF-MS, of which 8 proteins were upregulated such as actin beta, fibrinogen beta chain, ApoA-I ; and 6 proteins such as peroxiredoxin-2, glu-tathione S-transferase omega 1 were down-regulated when compared with controls. The result of ApoA-I by RT-PCR was consistent with the proteomics results. Conclusion Some differentially expressed proteins are observed in the PBMCs of patients with rheumatoid arthritis, which may play a potential role in the pathogenesis of RA.

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