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The aim of this study was to investigate the growth characteristics of primarily cultured astrocytes and microglia of different generations and then optimize the method for obtaining primary astrocytes and microglia effectively. Primarily cultured microglia were isolated and purified from the cortices of neonatal mice. The proliferation curve of mixed glia cells was measured by Cell Counting Kit-8 (CCK-8) assay, the proportion of astrocytes and microglia was detected by flow cytometry, and the polarization of the two types of glia cells was identified by immunofluorescence staining. Cell growth results showed that the mixed glia cells of P0 and P1 generation had the best proliferative activity; 97.3% of the high purity microglia could be obtained by mechanical shaking at 170 r/min for 30 min, and there was no significant difference in the morphology of ionized calcium-binding adapter molecule 1 (Iba-1) positive microglia and the proportion of M1 and M2 phenotype among the P0, P1 and P2 generations of microglia isolated by the above methods. Moreover, 95.7 % of the high purity astrocytes could be obtained by astrocyte cell surface antigen-2 (ACSA-2) magnetic beads separation, and there was no significant difference in the morphology of glial fibrillary acidic protein (GFAP) positive astrocyte and the proportion of A1 and A2 phenotype among the P0, P1 and P2 generations of astrocyte isolated by the above methods. Taken together, this study observed the growth characteristics of primarily cultured microglia and astrocyte in vitro, and then proved the best generations for purifying microglia and astrocytes. Finally, we optimized the methods of obtaining microglia and astrocyte, and verified that continuous culture within 2 generations will not affect the functional phenotypes of glia cells. These results provide technical support for studying the molecular mechanism of inflammation-associated diseases in nervous system.
Subject(s)
Mice , Animals , Astrocytes/metabolism , Microglia/metabolism , Cell Count , Flow Cytometry/methods , Cell Proliferation , Cells, CulturedABSTRACT
Based on the chemical structures of magnolol and honokiol,a series of small molecular derivatives were designed for the treatment of Alzheimer's disease.Through the Discovery Studio,five compounds (6a-6e) exhibited the inhibitory activity against Aβ and Tau proteins in all of the designed compounds.Then the five compounds are chemically synthesized and their biological activities were tested by thioflavin T.The result showed that compound 6a had inhibitory effect on the aggregation of two kinds of target proteins at the concentration of 100 μmol/L,which deserves further research.
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A soft coral Sinularia sp., collected from the South China Sea, was selected to investigate the bioactive and chemical constituents. The EtOAc fraction were isolated by repeatedly silica gel and Sephadex LH-20 column chromatography to obtain lobophytolide A (1), 3-dehydroxylpresinularolide B (2), sarcophine (3), 3 beta-acetoxyisolobophytolide (4), Crassocolide D (5), (3E,7E,11E)-6-acetoxy-3,7,11,15(17)-cembratrien-16,14-olide (6). The structures of compounds 1-6 were determined on the basis of spectroscopic data analysis. All compounds were tested against a small panel of human tumor cell lines. And these compounds were obtained for the first time from this coral.
Subject(s)
Animals , Anthozoa , Chemistry , Diterpenes , Chemistry , Molecular StructureABSTRACT
Biological mass spectrometry has been developed for the largE-scale protein identification. The successful identification of protein in proteomic study is based on an effective match of MS data to the sequence in database. Because of the diversity and heterogeneity of protein modification, the experimental data obtained by mass spectrometry does not match the theoretical value sometimes, which makes about 90 percent or more of the tandem mass spectra not be effectively identified. This has become one of the most important technique issues to be resolved in current proteome research. The N-terminal cyclization of peptides, as one of a variety of modification introduced in sample preparation, has been preliminarily studied in this work. The result showed that N-terminal cyclization occurred at the most of the glutamine(Q) or carbamoylmethyl-cysteine(CAM_C) residues and the reaction is often incomplete or partial, both types of peptides could often exist in its respective state at the same time, and the behavior of modified peptides in revered phase chromatography is also changed. The success rate of protein identification could be obviously improved if adding the N-terminal cyclization modification in the database searching. These results will be very helpful in the mass spectrometric data analysis of proteomic study.
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OBJECTIVE:To make clear the antimicrobials used for inpatients in our hospital from September 1999 to March 2000 METHODS:We sampled the records of discharged patients at the ratio of 1/3 of sickbeds in our hospital,and filled in "the questionnaire of antimicrobials used in clinical department" item by item,finally,we carried out comprehensive analysis RESU_LTS:More than half was administration of single and basic antimicrobial Combined use of antimicrobials for severe and mixed infections was rational The rate of using antimicrobials was high(79 64%),especially in preventing perioperative infections,reached nearly 100% The starting-point was high in choice of drug and the newly-developed,special and expensive drugs occupied the front place in order of consumption Combined use of drugs was unduly prevalent Most of treatment depended on experience The rate of bacterial drug resistance test was only performed in 10% of patients CONCLUSION:Rational use of drugs is the main trend,however,the measure directed against existing problem should be worked out
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OBJECTIVE:To select a best therapeutic scheme for diabetes among four kinds of oral hypoglycemic drugs:repaglinide(Novonorm),glibenclamide,gliclazide and glipizide.METHODS:According to the principle of pharmacoeconomics,using cost-effectiveness analysis(CEA),assessment and survey of the influencing factors(price,dosage)were carried out,at the same time,sensitive analysis was performed.RESULTS:The best therapeutic scheme was Novonorm and the rest were gliclazide,glipizide and glibenclamide in sequence.CONCLUSION:The result of sensitive analysis is in keeping with that of CEA,which indicates the credibility of CEA.
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0.05).CONCLUS_ION:The present study in rabbits showed no significant interaction between ciprofloxacin and tacrolimus after combined administration of these two drugs in a single dose.
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Objective: To compare nephrotoxicity and ototoxicity of gentamycin administered in single dose or multiple dose daily in guinea pigs. Methods: Thirty two male guinea pigs were divided into physiological saline control, single dose group daily (gentamycin, 120 mg/kg, 1/d) and multiple dose group daily (gentamycin, 60 mg/kg, 2/d). The physiopathology of renal and cochlea in guinea pigs were examined using auditory brainstem response (ABR), SC sound irritation and electron microscope. The gentamycin concentrations in serum and in perilymph were monitored by fluorescene polarization immunoassay (FPIA). Results: (1) Compared with control group, both gentamycin single and mulitiple daily doses injuried kidney and cochlea to some extent.The injury of multiple dose groups were worse than that the single dose groups ( P
