Study On Proteomics Of CSF In Central Nervous System With Infection And Dementia

(?) Study on proteomics of CSF in central nervous system with inflammation and dementia[Objectives] To investigate the citrullination difference in cerebrospinal fluid of sCJD patients and controls, in order to explore a potential biomarker in sCJD patients and better understand the pathogenesis and mechanism, so as to prevent and intervene on the disease. On the other point, we analyzed the PrPc?A?42 and Tau protein expression among the dementia and inflammation patients, to investigate the difference of those proteins expression. In order to find a relationship between dementia and inflammation cases, so as to explore a possible better treatment.[Methods] We conducted a case-control study.30 cases sCJD patients and 30 cases controls were enrolled. Use Western blot and 2D electrophoresis plus mass spectrum to detect citrullinated proteins in different groups, and later use Immunofluorescence double staining to localize the citrullinated proteins in brain sections. On the other subject,17 cases AD patients,12cases sCJD patients, 14cases encephalitis and 15 cases meningitis were enrolled, using Western blot and ELISA to check and analyze the different expression of PrPc, A?42 and Tau protein among those groups.[Results]1. Remarkable increase of citrullinated protein in CSF of sCJD is observed when comparable to the controls, which demonstrates the highly possible relationship between the pathogenicity of sCJD and abnormal increase of citrullinated proteins.2. The expression and activity of PAD2 in CSF of sCJD cases is significantly increased comparing to control group.3. Over-expressed citrullinated GFAP proteins was observed in CSF of sCJD cases.4. It was demonstrated by the double immunofluorescence that GFAP and citrullinated proteins are co-localized in frontal cortex of sCJD cases. The same result was also observed for PAD2 and GFAP proteins.5 The PrPc expression level in sCJD is 70% of control, in AD is lower than control but higher than sCJD group by Western blot. The PrPcexpression in inflammation group is the lowest among the above groups. PrPc value in sCJD is 138±75 ng/ml, in AD is 160±76 ng/ml by ELISA test, the control value is 225±6ng/ml.6 The A?42 expression level in sCJD, AD and inflammation groups are decreased compared to control. The decrease in sCJD group is remarkable, the level is nearly 40%of control by western blot analysis.7. The Tau expression level in sCJD and AD is remarkably increased comparing to control,the inflammation groups Tau level is also increased than control but lower comparing to sCJD and AD groups.[Conclusions]1. By using 2D-electrophoresis and MS analysis, the over-expressed citrullinated proteins in sCJD group are indicated as GFAP. Also we found citrullinated GFAP and citrullinated proteins are co-localized by immunoflurescence. Citrullination of GFAP is a possible potential biomarker for neurological degeneration disease.2. The higher expression and activity of PAD2 in sCJD group indicate that it plays an important role in GFAP citrullination.3. The concentration of calcium-ion plays the crucial role in the process of PAD2 catalyzed citrullination. Based on our results and literatures, it was concluded that high concentration of the calcium-ion in sCJD activated PAD2 enzyme, thus leading to GFAP citrullination.4. The relatively lower expression of PrPc proteins in sCJD group, AD and inflammation compared to the controls suggests that certain underlying mechanism related to PrPc proteins exit between the CNS degeneration disease and CNS inflammation disease.5. The lower expression of A?42 in sCJD AD inflammation group than control reflecting the relationship between the CNS degeneration disease and CNS inflammation disease.6. The expression level of Tau protein in sCJD group, AD and inflammation is higher compared to controls, among which the sCJD group is the highest. (?)In vitro cytotoxicity of monodispersed hematite nanoparticles on Hek 293 cells[Background]With its rapid development, nanotechnology has found its way into our daily life step by step, and subsequently more and more nano materials emerged around the surrounding environment. Due to the unique physical and chemical performs resulting from the quantum size effect, nano materials show promising applications in many fields, such as chemical engineering, ceramics, microelectronics, metrology, electrics, optics and information communication system. Furthermore, recent studies demonstrated its potentials in biology and medicine, including disease diagnosis, molecular imaging, and fluorescence marker for bio-sensor, transmission of drugs and genes, and tissue engineering. As a result, human beings are subject to be exposed in nanomaterials environment in various ways, for example, breath, eating and skin touching. After invading into the human body, nanomaterials will disperse in different organs, and even penetrate through the bio-barriers. More and more attentions have been paid to the biotoxicity and bio-compatibility of nanomaterials interacting with human beings and environment. Enhancing the investigation of the underlying toxicity and interaction mechanism of nanomaterials on human beings and environment would facilitate helping us enjoy the great advantages nanotechnology bringing to us, meanwhile avoid the potential harm from the nanomaterials to a high degree.[Aims&Methods]Iron oxides are widely used in our daily life, on the one hand, as pigment, catalyst, ceramics and energy conversion materials, and on the other hand, as an important candidate of magnetic materials for smart target therapy and drug carrier in medical field. However, good bio-compatibility without obvious biotoxicity is required for these applications. Inspired from these demands for the practical applications, this work focuses on the investigation of biotoxicity effect and the principle mechanism ofiron oxide nanoparticles on the human HEK293 cells with the MTT colorimetry, activity of SOD and GSH, LDH and MDA measurements, aims to provide the basic knowledge for evaluating the advantage and disadvantage of the nanoparticles in medical application.[Results]Following results were demonstrated based on the various measurements. Dosage-and time-dependent biotoxicity was observed for HEK 293 cells when exposed in iron oxide nanoparticles as-synthesized in the context. Both large dosage and long-term exposure in nanoparticle environment can lead to the serious cell toxicity. SOD and GSH-Px activity decrease for all dosages of hematite nanoparticles, thus leading to the increase of LDH leaking and MDA concentration.[Conclusions]In summary, some conclusions were reached as follows:(1) Iron oxide nanoparticles display the dosage-and time-dependent biotoxicity on the HEK 293 cells. (2) Reactive oxygen freeradicals and oxidative stress were proposed to be the dominant biotoxic mechanism in this study. The interaction between nanoparticles and cells leads to the increase of reactive oxygen freeradicals such as superoxide anion, hydroxyl freeradical and peroxide, which breaks through the anti-oxidization barrier and finally causes the damage of epicyte and the apoptosis of cells.