ABSTRACT
Objective As SELDI-TOF-MS (Surface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry) has been broadly used to screen biomarkers for a variety of diseases, the identification and validation of the revealed biomarkers requires more focused attention.Method In this paper, the serum samples from 60 cholangiocarcinoma, 146 lung cancer, 65 LGC and 58 LPC, 49 benign diseases of hepatobiliary and 53 normal individuals were analyzed by SELDI-TOF-MS. Results Among a set of proteins automatically selected as specific biomarkers by Biomarker Wizard software, three protein peaks, with molecular weights of 13. 71 × 103 , 13.83 × 103 and 13. 99 ×103 , were found significantly decreased in cholangiocarcinoma samples. The candidate biomarkers obtained from Tricine-SDS-PAGE gel bands by matching the molecular weight with peaks on CM10 chips were identified by Mass spectrometry as the native transthyretin(native TTR),cysTTR and glutTTR.These preliminary results were further proven by immunoprecipitation using commercial TTR antibodies. This allowed us to re-measure the TTR levels in all the groups more simply by ELISA assay. It showed a firm consistency between ELISA and SELDI analysis. In addition, while TTR levels in cholangiocarcinoma were found to be lower than those in normal healthy controls, TTR levels in benign diseases of the hepatobiliary system were found to be higher than those in healthy controls.Conclusion TTR could be a biomarker that better discriminates cholangiocarcinoma patients from the benign diseases compared to other biomarkers presently available.
ABSTRACT
Objective Because in the environment there are many compounds to cause variation of cells, the study for high sensitivity’s method in which the variation of cells by environment can be tested is necessary. For example, the reports of detailed study for the application of cadmium on the human body cell’s lethal toxicity and the variation of cells are very few. This research is about a high sensitivity method of heavy metal cadmium’s variation active processed by cadmium in which the variation of cells can be tested by ultraviolet ray. Methods The study used cultured cells of human being and derivation. The assay of mutagenicity OuaR (Ouabain resistance) variation used picking law and Differential dot-blot of cancer gene K-ras variation the hybridization examination method. In order to test the induction’s frequency of cell variation, MTT (methyl thiazolyl tetrazoliun) and survival efficiency of formation were tested. The clarifing of mutagenicity mechanism used Western blot and the siRNA method to analysis the molecular of heat shock protein. Results Cadmium clarifiing mutagenicity variation detection had the same degree with the method of ultraviolet ray. The DNA was injury corresponding to the HSP27 protein cell’s change.The cell variation frequency also presented the high value. Conclusion Human fibroblasts in the role of the heavy metal cadmium showed that HSP27 protein expression in high-UVr-1 cells RSa has lowor-frequency variations than OuaR cells. K-ras dot blot method proved that RSa cells variation is higher than the cells UVr-1 mutation. In siHSP27 cells, we observed OuaR high variability of expression. Cadmium on DNA damage showns UVr-1 fibroblast cell is lower than RSa fibroblasts cell.
ABSTRACT
The BHK-21 (baby harmster kidney cells) growing on glass are observed by SEM after critical point drying. The cell shape, the main surface structure, and their alteration during the cell cycle are described. As a particular stage, the senile cells are also mentioned.Three other cell lines: HeLa, IB-RS-2 and chicken fibroblast, sampled in same way, are studied for comparison. In all these four cell lines the changes of cell surface morphology follow a common pattern. And notably, the microvilli of all of them have the similar diameters, though their shapes and distributions are distinct from each other.Using cytochalasin B and colchicine as specific inhibitors it is shown that the microvilli and blebs are mainly related to the microfilaments, while the filapodia are to microtubules.
ABSTRACT
Objective To establish an efficient, simple, low cytotoxicity and cheap transfaction system. Methods We have used the cationic polymer polyethylenimine(PEI) to study transient transfection in MCF-7 cells by testing different conditions, including cell concentrations, DNA concentrations, the ratio of PEI nitrogen to DNA phosphate(PEI-N∶DNA-P) and the time of cells grown in serum-free culture together with PEI-DNA complex. Results The optimized cell concentrations were 2?10 5 cells seeded per well in 24-well dishes 18-24?h before transfection. The DNA concentrations and ratio of PEI-N∶DNA-P are very important for optimal transfection and they affect each other. For 1??g DNA per well, the optimal PEI-N∶DNA-P is about 33∶1, however, as for 4??g DNA, it is 9∶1. The best time of cells grown in serum-free culture together with PEI-DNA complex is about 5-7?h.Conclusion With optimized conditions, we can establish an efficient, simple, low cytotoxicity and cheap transfection system by using PEI.