2.Overexpression of 14-3-3 protein alleviates the toxicity of MPP~+ to PC12 cell
Xiao-Wu CHEN ; Sheng-Gang SUN ; Dao-Bin CHENG ; You-Yong TIAN ; Jing HE ;
Chinese Journal of Neurology 2001;0(02):-
Objective To investigate the protective effects of the 14-3-3 protein overexpression on the injury of PC12 cell induced by MPP~+ and its mechanisms.Methods For expression in mammalial cells, pcDNA3.1(+)-14-3-3 plasmid was constructed and transfeeted into PC12 cell with Lipofectamine~(TM)2000. The overexpression of transfected 14-3-3 gene in PC12 cell was determined by immunofluorescence and Western blotting.The effects of 14-3-3 overexpressing on the cells viability,apoptotie ratio and the activity of superoxide dismutase(SOD)as well as glutathione peroxidase(GSH-Px)of PC 12 cell treated with MPP~+ were measured by MTT assay,flow cytometry analysis and microplate reader respectively.Results The expression of 14-3-3 protein in transfection group(1.19?0.06)increased evidently compared with control group(0.75?0.05).And the antioxidant enzyme activity assession,MTT assay and flow cytometry analysis shows that the overexpression of 14-3-3 protein elevates the activity of SOD(transfection group:(9.13? 0.41)U/mg protein,MPP~+ group:(6.45?0.52)U/mg protein)and GSH-Px(transfection group: (89.66?3.42)?mol/mg,protein MPP~+ group:(82.73?4.15)?mol/mg protein),increases the cell viability(transfection group:0.78?0.06,MPP~+ group:0.54?0.07),and inhibits cell apoptosis (transfeetion group:11.87%?3.26%,MPP~+ group:36.30%?2.39%)of PC12 induced by MPP~. Conclusion The overexpression of 14-3-3 protein could elevate the activity of antioxidant enzymes SOD and GSH-Px,reduce oxidant stress,alleviate MPP~+ toxicity,and thus inhibit the apoptosis of PC12 cell induced by MPP~+.
3.Detection of a novel glyoxalase in human adipocytes and analysis of its structure and function
Xiao-Hua LI ; Sheng ZHENG ; You-Ping LIU ; Tian-Hong LUO ; Min LUO ;
Chinese Journal of Endocrinology and Metabolism 1986;0(03):-
Objective To identify novel proteins of human adipocytes by proteomic study,and analyze their strueture and function.Methods The expressed proteins of human adipocytes were identified by 2-D electrophoresis and mass spectrometry,and the structure and function of the novel proteins were predicated via bioinforrnatics.Results My027 is a novel protein of human adipocytes identified by 2-D electrophoresis and mass spectrometry.This protein was confirmed by RT-PCR,sequencing,and prokaryotic expression.My027 protein is located in the cytoplasm and has highly conserved structure,indicating that it is an important functional protein. This protein is highly homologous to glyoxalase superfamily with a whole domain of glyoxalase.It shares 96.7% homologous to glycoxalase between 141-253 amino acids and 95.7% homologous to glycoxalaseⅠbetween 144-244 amino acids.Conclusion The results show that protein My027 appears to have the same function as glyoxalaseⅠ.
4.Isolation and Identification of Osmotolerant Yeast from "Swollen Can" Soy Sauce
You-Sheng OUYANY ; Xiao-Bao XIE ; Jiao-Di CHEN ; Hua XUE ; Yi-Ben CHEN ;
Microbiology 1992;0(04):-
Three osmotolerant yeasts were isolated from three batches of "swollen can" soy sauce produced by a Guangdong condiment plant. These strains grew faster in the media containing 50%~60% glucose or 15% NaCl than in common yeast media. The three yeasts were identified as Pichia etchellsii by using morphological characteristics, physiological and biochemical tests.
5.The Microorganism Constitutes Analysis of Soy Sauce Liquor from High-Salt-Level Watery State Fermentation
Xiao-Bao XIE ; You-Sheng OUYANG ; Hai-Yan ZENG ; Chun-Hua WANG ; Yi-Ben CHEN ;
Microbiology 1992;0(03):-
A system analysis about the microbial flora of normal and abnormal soybean sauce liquor from the high-salt-level watery state fermentation was made and the dominant bacteria and yeasts were identified.On the other hand,a discussion of effect of temperature on microbial flora was made.The results indicated that there were no obvious differences about the count of aerobe,spore-producing bacteria,enterobacteriaceae,lactic acid bacteria and anaerobe between the normal and abnormal soybean sauce liquor and there were marked differences about the count of yeasts and salt-tolerant bacteria.The predominant yeasts in normal soybean sauce were Torulopsis and Saccharomyces,accounting for 55.9% and 35.3% of the total yeasts separately,and in abnormal soybean sauce were Pichia,candida and Saccharomyces,accounting for 62.8%,17.9% and 9.0% respectively.
6.The Developments of Microbial Resistance to Industrial Antiseptics and Disinfectants
Chun-Hua WANG ; Xiao-Bao XIE ; Hai-Yan ZENG ; You-Sheng OUYANG ; Yi-Ben CHEN ;
Microbiology 1992;0(04):-
These years with industrial antiseptics and disinfectants widely used, especially unscientifically used, the resistance of microorganisms is more and more serious, which brought much more difficulties to industrial producing. The developments on the resistance of microorganisms to industrial antiseptics and disinfectants, their resistant mechanisms and the control strategies are reviewed, which aids us to reasonably use industrial organic antiseptics and disinfectants in existence and provides academic and scientific basis.
7.Study of the follow-up management strategies after installation of improved stoves to prevent coal-burning type endemic fluorosis in Guizhou province
Bo-you, ZHANG ; Da-sheng, LI ; Yin, LIANG ; Xiao-qiang, HU ; Rui-zhi, ZHANG
Chinese Journal of Endemiology 2011;30(6):697-700
Objective To study the follow-up management strategies after improving stoves for controlling coal-burning type endemic fluorosis in Guizhou and to provide a scientific basis for exploring the follow-up management measures.Methods In 2006 - 2009,three counties of Puding,Bijie and Liuzhi with improved stoves in 2005 and implemented follow-up management measures for subsequent three years were chosen,3 towns were chosen randomly in each chosen county,2 villages were selected randomly in each chosen town,10 households were investigated randomly in each chosen village,and 20 students were investigated of the knowledge of fluorosis control.The investigation included also the usage of the stoves,dehydration methods of grain and related conditions.Results ①The head of the family and student awareness of fluorosis control were 82.6%(743/900)and 91.7% (1650/1800),respectively,and 35.0% ( 385/1100 ) and 61.0% (6605/10 835 ),respectively,before the test,and the difference was statistically significant( x2 =33.04,1189.12,all P < 0.01 ).②The rates of chimney out of the house were 88.5%(69/78) and 100.0%(102/102),respectively,compared with those before the trial[74.4%(2125/2856) and 2.1%(104/4984),respectively],the difference was statistically significant(x2 =720.56,4295.38,all P < 0.01).The iron stove and the table stove's airtight utilization rates were 85.9%(67/78) and 100.0%(102/102),respectively.③The corn and the hot pepper's correct drying rates were 100.0%(180/180).Compared with those[27.2%(49/180) and 32.2%(58/180),respectively]before the trial,the difference was statistically significant(x2 =26.68,37.38,all P < 0.01 ).The corn and the hot pepper's washing rates before eating were 95.0%(57/60) and 98.3%(177/180),respectively.Compared with those[85.0%(153/180) and 77.8%(140/180)]before the trial,the difference was statistically significant(x2 =135.00,490.82,all P < 0.01 ).Conclusions The target population's awareness and knowledge of fluorosis prevention and related behavior are significantly elevated and enhanced after implementation of the three years post-management,which has reached the desired goal.The strategies of the follow-up management suits Guizhou province and widespread application is suggested.
8.The Concentration and Variation of Airborne Microbe in Guangzhou City
You-Sheng OUYANG ; Xiao-Bao XIE ; Yi-Ben CHEN ; Xiao-Mo HUANG ; Hong PENG ; Qing-Shan SHI ;
Microbiology 1992;0(03):-
One year survey on the concentrations and monthly or seasonal variations of airborne microbe in Guangzhou city were analysed and studied with JWL-IIB airborne microbial sampler. The results showed that the yearly average airborne microbe content of outdoor was 2, 298 cfu/m3, and that of indoor was 1,792 cfu/m3 in Guangzhou city. The monthly variation range of outdoor airborne microbe was from 1,073 to 4,096 cfu/m3, the highest content was 4,096 cfu/m3 in March, and the lowest content was 1,073 cfu/m3 in October. The outdoor airborne bacteria and fungi counts were the highest in spring, next in summer, lower in winter and the lowest in autumn in the four seasons . The yearly average concentrations of outdoor airborne microbe at the Garbage compression station, the business walk street, the key traffic route, the residential area, the industrial district and the garden were 4, 573, 3, 835, 1, 580, 1,413, 1, 197 and 1, 187 cfu/m3respectively; and Ones of indoor at the key traffic route, the tourist three star-route hotel and the subway station were 2,511, 1,699and 1,167 cfu/m3 respectively . The study on airborne microbe can be used for the research of health prevention and environment control measures in Guangzhou .
9.Isolation and Identification of BLN-2 Producing Poly-?-Glutamic Acid and Studies on Its Solid-state Fermentation
Jing FENG ; Qing-Shan SHI ; Xiu-Lin SHU ; Xiao-Ping LIN ; You-Sheng OUYANG ; Yi-Ben CHEN ;
Microbiology 2008;0(09):-
A poly-?-glutamic acid producing strain--BLN-2, was isolated from the soybean products. According to the biochemical characteristics and 16S rRNA, the strain was identified as Bacillus subtilis. Using soybeans as culture, the solid-state fermentation conditions of BLN-2 have been studied. The results showed that the optimal carbon and nitrogen sources of BLN-2 were glucose, fructose, NaNO3 and KNO3, respectively. The orthrogonal experiments showed, when the final concentration of the fructose which was added to the soybean culture was 0.5%, the glucose, NaNO3 and KNO3 final concentraion were 2.0%, the production of ?-PGA was the highest--89.05 g/kg. It is 48.42% higher than other comparable soybean medium under the same conditions.
10.Prokaryotic Expression and Acetylation Assays of Histone Acetyltransferase PCAF
Wei-Zhong LI ; Dan-Gui ZHANG ; Jun ZENG ; Ge-Fei WANG ; Xiao-Xuan CHEN ; You-Ying CHEN ; Kang-Sheng LI ;
China Biotechnology 2006;0(01):-
P300/CBP-associated factor(PCAF),an important member of histone acetyltransferase family(HATs) within eukaryotic cells,is capable of inducing the acetylation of histone,promoting the transcription of specific genes and involving in many biological effects.In the present study,full-length cDNA of PCAF was inserted into plasmid pGEX-5x-1,then the soluble protein GST-PCAF was expressed in E.coli BL21(DE3) after the optimization of inducing conditions.The recombinant protein was further purified with affinity chromatography and tested the activity by in vitro acetylation assays.High efficient PCAF protein produced by this method could serve for the study on the role of PCAF in gene regulation and the interaction between PCAF and other proteins.