1.MAR-FISH Technique and Its Application in Study of Environmental Microbial Community and Function
Xiao-Hui WANG ; Xiang-Hua WEN ;
Microbiology 1992;0(01):-
The major goal of microbial ecology is to study the structure and function of complex micro-bial communities. New molecular biological techniques have been successfully applied to analyze mi-crobial community structure. However they do not provide information on the physiologic properties of the detected microorganisms. A new tool for structure-function analyses in microbial ecology, micro-autoradiography combined with fluorescence in situ hybridization (MAR-FISH) can be used to simul-taneously examine the phylogenetic identity and the specific activity of microorganisms within a com-plex microbial community at a single-cell level. This article reviews the principle, experimental steps of MAR-FISH technique. The application of this technique in study of the environmental microbial com-munity and function is also summarized.
2.Transfusion transmitted virus infection in the neonates
Wen-Xiang WANG ; Ai XIONG ; Xiaoxin HUA ;
Chinese Journal of Infectious Diseases 2001;0(03):-
Objective To investigate the relationship between transfusion transmitted virus (TTV)infection and neonatal hyperbilirubinemia,the effect of TTV infection on the liver function, and analyse the feature of nucleotide sequences in TTV ORF1.Methods Serum TTV DNA,which were from 58 neonates with high direct bilirubin(DB,including 5 with hepatitis Syndrome),92 ones with high indirect bilirubin(IB),and 85 normal ones,was detected using a nested polymerase chain reaction technique(nPCR),electropherosis and sequence analyse,and serum alanine amniotransferase (ALT)was determined in all neonates.Results In DB neonates,TTV DNA were detected in 7 neo- nates(12.1%,including 3 neonates with hepatitis syndrome);in IB and normal ones,1 neonate had positive TTV DNA(1.1% and 1.25),respectively.Even if there were point mutations in Guangdong's TTV,the homology of Guangdong's TTV(GD1-9)and Japanese TTV(N22)ranged from 87.1%~97.8% at nucleotide level.Conclusion TTV infection may be one of important pathogenesis resulting in neonatal hyperbilirubinemia and liver damage in such patients.Guangdong's and Japanese TTV isolates had the same genotype,some gene mutations maybe increase the pathogenicity in TTV.
3.Reference methods and reference measurement principles in clinical biochemistry
Wen-Xiang CHEN ; Zi-Yu SHEN ; Zhen-Hua YANG ;
Chinese Journal of Laboratory Medicine 2001;0(03):-
Characteristics and measurement principles of reference methods in clinical biochemistry were described.Implementation of reference systems is one of the most effective approaches to improve the accuracy and comparability of clinical laboratory test results.Reference methods are the key components of reference systems.Reference methods should have measurement uncertainties that meet the requirements of the intended use,and thus should be based on reliable measurement principles.For the well-defined biochemistry analytes,reference methods have been almost all based on instrumental analysis.Isotope dilution mass spectrometry (ID/MS) is considered most reliable and has been the major analytical principle of the reference methods.ID/MS analysis is accurate but expensive.Use of other validated instrumental analyses as reference measurement principles would be justified.
5.Protection of extract of Radix Atragali composite against acute hepatic injury
Hua XU ; Jie WEN ; Liping YU ; Xiaoming YANG ; Jizhou XIANG
Chinese Journal of Pathophysiology 1986;0(02):-
AIM: To study the protection of extract of Radix Atragali composite against acute hepatic injury. METHODS: Fed with the extract of Radix Atragali composite, m ice were injected with D-galactosamine intraperitoneally (800 mg/kg) and rats were i njected with carbon tetrachloride hypodermically (5 mL/kg) to induce acute hepat ic injury on the 8th day. ALT, AST and bilirubin in serum were examined. Patholo gical changes of liver tissue were observed. RESULTS: Compared with model group, activities of ALT and AST, c oncentrations of bilirubin were markedly decreased and pathological scores also showed that degeneration and necrosis of hepatic cell were lighter in the the ex tract of Radix Atragali composite treatment group. CONCLUSION: The extract of Radix Atragali composite attenuat es hepatic injury induced by D-galactosamine or carbon tetrachloride.
6.Screening of High Daptomycin-producing Strain by He-Ne Laser Irradiation and Streptomycin Resistance Screening Method
Wen-Yu LU ; Jian-Ping WEN ; Jing-Hua FAN ; Bo-Xiang CAO ; Bing SUN ;
Microbiology 1992;0(03):-
The spores suspension of Streptomyces roseosporus D-38 irritated with 20mW He-Ne laser for 20 min were incubated on G1 medium plates containing 1. 9?g/mL of streptomycin. Ten percent of mutants increased the potency of daptomycin by streptomycin-resistance method, including the mutant LC-54, which could produce daptomycin 81. 2 mg/L, which was 39% higher than that of the beginning strain by flask fermentation.
7.Dimerization of retroviral RNA genomes.
Xu GAO ; Rong-Xian SHEN ; Wen-Hua XIANG ; Jian-Hua ZHOU
Chinese Journal of Virology 2008;24(6):487-491
Base Pairing
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Dimerization
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Genome, Viral
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RNA, Viral
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chemistry
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genetics
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Retroviridae
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chemistry
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genetics
8.Effects of mycorrhizal colonization and medicine quality of Paris polyphylla var. yunnanensis inoculated by different foreign AM fungi species.
Zhou NONG ; Bo DING ; Yuan FENG ; Wen-hua QI ; Hua ZHANG ; Dong-qin GUO ; Jun XIANG
China Journal of Chinese Materia Medica 2015;40(16):3158-3167
After 28 foreign species of AM fungi were inoculated in sterilized soil, the effects of the AM mycorrhizal colonization and the medicine quality of Paris polyphylla var. yunnanensis were observed by combination of inoculation test in pot at room temperature and instrumental analysis. The results showed that, compared with control group (CK), the inoculation of foreign AM fungi in the soil influenced the spore density, mycorrhizal infection rate, and colonization intensity of AM fungi in root system of P. polyphylla var. yunnanensis. The inoculation of foreign AM fungi enhanced the mycorrhiza viability of P. polyphylla var. yunnanensis by increasing the activity of succinic dehydrogenase (SDH) and alkaline phosphatase (ALP) in intraradical hyphae. The content of single steroid saponin in rhizome of P. polyphylla var. yunnanensis showed variation after P. polyphylla var. yunnanensis was inoculated by different foreign species of AM fungi, which was beneficial for increasing the medicine quality; however, the kinds of steroid saponin showed no difference. In a degree, there was a selectivity of symbiosis between P. polyphylla var. yunnanensis and foreign AM fungi. And we found that the Claroideoglomus claroideum and Racocetra coralloidea were best foreign AM fungi species for cultivating P. polyphylla var. yunnanensis under field condition.
Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal
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analysis
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Fungi
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classification
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growth & development
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Liliaceae
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chemistry
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growth & development
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microbiology
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Mycorrhizae
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classification
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growth & development
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Plant Roots
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chemistry
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growth & development
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microbiology
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Quality Control
9.Identification of serum biomarkers for rheumatoid arthritis using protein fingerprint
Wen-Bo LIU ; Xing-Fu LI ; Feng DING ; Hua-Xiang LIU ;
Chinese Journal of Rheumatology 2000;0(06):-
Objective To identify serum biomarkers for rheumatoid arthritis(RA)by protein finger- print pattern. Methods One hundred and forty-one serum samples of 90 RA patients, 20 systemic lupus ery- thematosus(SLE)patients, and 31 healthy individuals were randomly divided into training set(n=93, 60 RA patients, 13 SLE patients and 20 healthy individuals)and test set(n=48, 30 RA patients, 7 SLE patients and 11 healthy individuals). They were detected by surface enhanced laser desorption/ionization-time of flight- mass spectrometry(SELDI-TOF-MS). The protein fingerprint pattern obtained from SELDI-TOF was trained by a multi-layer artificial neural network(ANN)to establish a diagnostic model. Results The detective mod- el obtained by ANN was used to detect the 48 unknown serum samples. The sensitivity and specificity for RA detection was 90% and 90.9% respectively. Conclusion In comparison with traditional methods, SELDI- TOF-MS could identify new serum biomarkers in RA. Combined with ANN, it provides high sensitivity and specificity for RA diagnosis.
10.Two-step Tandem Chromatography Purification of Anti-human CD80 Monoclonal Antibody 4E5 from Mouse Ascites
Hong-Bing MA ; Yu-Hua QIU ; Ran TAO ; Wen-Xiang LI ; Ying XU ; Xue-Guang ZHANG ;
China Biotechnology 2006;0(08):-
A two-step method for the purification of blocking-type anti-human CD80 monoclonal antibody 4E5 from mouse ascites was developed using anion exchange and gel filtration in combination. The ascites was first purified by anion exchange after centrifugation and filtration. The experimental parameters of sample loading and elution were optimized. The optimized loading condition was pH 8.0,50 mmol/L Tris-HCl and satisfactory results were obtained using a 0~0.5mol/L NaCl step elution. The fraction containing the protein of interest was directly loaded on gel filtration column and eluted using a 20 mmol/L phosphate buffer at pH 7.2. The purity of the obtained monoclonal antibody was up to 95% with a recovery of 61%. The purity of mAb could efficiently inhibit the growth of Daudi cells. The amplification of the method was also studied using a Bio-Scale Q5 column and the result was satisfied.