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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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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.
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In this article,stereologic method was applied in analyzing 17 morphological parametera of cell, nucleus and nucleolus in 20 cases of mid stage and late stage of the simple carcinoma of breast (the stylebooks of EM photos were 320). We analyzed the difference of each sort of cell with T test and Q test. Compared with the normal cells, the cancer cells of mid stage were found obviously not only decreased in their nucleus surface density Svn and nucleus contrast surface density Sn but also increased in their nucleus average section An and nucleus average perimeter Bn. Compared with the cancer cells of the mid stage the cancer cell of the late stage were found obviously increased in their nucleus average volume Vn and Bn. Compared with the normal cells, the cancer cells of the late stage were found obviously not only decreased in their Svn and Sn but also increased in their Vn, An, Bn and nucleolus average perimeter Bu.
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Purpose:To study the expression of P53 and its relationship with hepatitis B virus(HBV) infection in hepatocellular carcinomas (HCC) and chronic liver diseases.Methods:HBsAg, HBcAg and P53 were detected by immunohistochemical staining and HBV DNA by in situ hybridization with total 98 cases of chronic hepatitis, liver cirrhosis, paratumor cirrhosis , HCC and normal liver tissue. Results:The positive rates of HBsAg, HBcAg and HBV DNA were 61.9%(13/21),42.9%(9/21) and 75.0%(12/16) in chronic hepatitis;64.0%(16/25),36.0%(9/25) and 83.3%(15/18) in liver cirrhosis; 72.7%(16/22),61.1%(11/18) and 85.7%(12/14) in the paratumor cirrhosis,as well as 45.2%(14/31),50.0%(14/28) and 64.3%(9/14) in HCC . The positive rate of P53 protein was 27.9%(12/43) in HCC. It was negative in the other groups.Conclusions:The expression of P53 occurs in the later stage of HCC .The infection of HBV is not related to the positive rate of P53 protein in HCC.