1.Significance of ~(99m)Tc-Diethyl Iminodiacetic Acid Hepatobliliary Planar and Tomographic Scintigraphy in Diagnosis on Biliary Atresia
ji-gang, YANG ; da-qing, MA ; chun-lin, LI ; lan-fang, ZOU
Journal of Applied Clinical Pediatrics 2006;0(19):-
Objective To evaluate the clinical value of 6 h 99mTc-diethyl iminodiacetic acid (99mTc-EHIDA) planar hepatobiliary scintigraphy (HBS),6 h tomographic HBS and 24 h planar HBS in diagnosis on biliary atresia(BA).Methods Seventy cases(32 male,38 female) with continuous jaundice received planar and tomographic HBS in Beijing Friendship Hospital from Jan.2005 to Dec.2007.The mean age was 48.7 d (29 d-4 months).According to final diagnosis,all cases were divided into BA group (45 cases) and non-BA group (25 cases).All cases fasted at least 4 hours before HBS.The equipment was 3 head IRIX from Philips company with low energy high resolution collimator.The tracer was 99mTc-EHIDA and the radiochemistry purity was more than 95 percent.The dosage was 7.4 MBq/kg.All diagnosis demonstrated by operation pathology and clinical follow-up.All cases received HBS at 5,10,15,20,30 min and 1,6 h after tracer injection.HBS would ended if radioactivity appeared in gallbladder or intestine.These cases would received tomographic HBS and 24 h HBS if radioactivity did not appear in gallbladder or intestine at 6 h post injection.All these images were analyzed by 2 or more nuclear medicine physicians.Results There were not radioactivity appearing in gallbladder and intestine on planar and tomographic HBS of 27 cases,which suggested the BA.There were radioactivity appearing in gallbladder and intestine on planar and tomographic HBS of 30 cases,which suggested the non-BA.Positive rate of 6 h tomographic HBS was significantly higher than that of 6 h planar HBS and there was significantly difference between the 2 methods.Positive rate of 6 h tomographic HBS was significantly higher than that of 24 h planar HBS and there was significant difference between the 2 methods.Conclusions 99mTc-EHIDA HBS is a noninvasive,safety,valuable examing method and has definitely clinical value in the diagnosis on BA.The clinical value of 6 h tomographic HBS is significantly higher than that of 6 h planar HBS and 24 h planar HBS.
2.Cloning, Expression and Property Analysis of Arabinosidase in Pichia pastoris
Dong-Sheng YAO ; Hui-Mei TAN ; Hui HUANG ; Da-Ling LIU ; Chun-Fang XIE ;
China Biotechnology 2006;0(04):-
SMART-RACE was performed after isolating the total RNA of Armillariella tabescens to amplify the full-length cDNA of arabinosidase (GenBank Accession No. AJ620046). Bioinformatics analysis was used to analyze the code frame of arabinosidase, to predict its structure and function. Recombinant plasmid pPIC9-AF was constructed and then electroporated into methylotrophic yeast Pichia pastoris GS115. The secreted 6 ? His fusion protein was purified to analyze its enzymology property. This arabinosidase had high activity at 30-35℃ under acid condition, and was stable within wide range of pH and temperature. It maintained about 80% activity at the range of pH4. 0-8.0 and 20-40℃,wider than many other cloned arabinosidase. So it was worthy to go step further to study this enzyme, and recombinant expression provided a chance of highly expressing arabinosidase.
3.Inducement, Purification and Characterization of?-mannanase from Armillariella tabescens EJLY2098
Dong-Sheng YAO ; Xiao-Kui HUANG ; Da-Ling LIU ; Chun-Fang XIE ; Rong HU ;
China Biotechnology 2006;0(07):-
Armillariella tabescens EJLY2098 was capable of secreting p-mannanase by konjac inducement. A 34 orthogonal design was applied to determine the optimum medium of inducing mannanase by Armillariella tabescens EJLY2098. The results suggested that Armillariella tabescens EJLY2098 secreted the high-activity enzyme in the optimum medium, which was composed of 2% konjac, 1% peptone, 25% potato juice,0.3% KH2PO4,15% MgSO4?7H2O, 0.01% VitB1. Purified by DEAE-anion exchange chromatography, two eluting peaks (P1 and P2) with the p-mannanase activity were obtained, and one of them (named?-mannanase P2) was a single band by the SDS-PAGE, and the molecular weight of?-mannanase P2 was 78. 9kDa. The isoelectric point of?-mannanase P2 was estimated to be 4.0-4. 1. The optimum activity for the enzyme was found at 60℃and pH4. 0 - 6. 0, and the enzyme was stable between pH4. 5 - 6. 0. The activity of?-mannanase P2 were enhanced by Na+ and Ba2+ . This?-mannanase can be used in feed industy. a new fungi secreting?-mannanase was obtained, providing an important base for cloning mannanase gene and constructing recombin microbe expressing?-mannanase .
4.Preliminary Study on Solid-state Fermentation of Streptomyces Strain Men-myco-93-63
Chun-Ying HAO ; Qing-Fang MENG ; Guo-Tong CHI ; Wen-Xiang YANG ; Da-Qun LIU ;
Microbiology 2008;0(09):-
The culture medium and cultural conditions of solid-state fermentation of Streptomyces Menmyco-93-63 were tested in this study. The suitable medium which contains rice, sorghum, millet bran, and rice hull with the proportion of 2:2:3:3 was developed for the spore production of Streptomyces Men-myco-93-63 using single substrate screening, mixture substrate screening and orthogonal experiments, and the sporulation was up to 2.52?109 CFU/g. And then, initial charge, initial ratio of water to solid, inoculating quantity, and culture temperature impact to sporulation of Streptomyces Men-myco-93-63 were tested. The favorite cultural conditions are developed as the following: the initial charge is 15 g in 500 mL Erlenmeyer flask; initial ratio of water to solid is 1.7:1.0 (V/W, rice hull excluding), inoculating quantity is 7 mL, culture temperature is 28℃.
5.Role of transporters in hepatic drug disposition.
Chun-Ying GAO ; Xiao-Yan CHEN ; Da-Fang ZHONG
Acta Pharmaceutica Sinica 2012;47(5):565-572
Liver is regarded as one of the most important organs for drug clearance in the body, which mediates both the metabolism and biliary excretion of drugs. Transporters are a class of functional membrane proteins and control the movement of substances into or out of cells. Transporters, which are extensively expressed in the liver, play important roles in the drug hepatic disposition by regulating the uptake of drugs from blood into hepatocytes or the efflux of drugs and their metabolites into bile. In this review, the localization, functions and substrate selectivity of the major transporters in the liver will be summarized, and the impacts of these transporters on drug hepatic disposition, the potential drug-drug interactions as well as their genetic polymorphisms will also be reviewed.
ATP Binding Cassette Transporter, Sub-Family G, Member 2
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ATP-Binding Cassette Transporters
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genetics
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metabolism
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ATP-Binding Cassette, Sub-Family B, Member 1
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genetics
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metabolism
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Bile
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metabolism
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Biological Transport
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Drug Interactions
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Humans
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Liver
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metabolism
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Membrane Transport Proteins
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genetics
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metabolism
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Metabolic Clearance Rate
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Multidrug Resistance-Associated Proteins
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genetics
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metabolism
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Neoplasm Proteins
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genetics
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metabolism
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Organic Anion Transporters
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genetics
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metabolism
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Organic Anion Transporters, Sodium-Dependent
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metabolism
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Organic Anion Transporters, Sodium-Independent
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genetics
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metabolism
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Organic Cation Transport Proteins
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genetics
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metabolism
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Pharmacokinetics
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Polymorphism, Genetic
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Symporters
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metabolism
7.Construction of a cDNA Library and Cloning of an Arabinosidase cDNA from Armillariella tabescens
Dong-Sheng YAO ; Hui HUANG ; Long ZHAO ; Chun-Fang XIE ; Da-Ling LIU
China Biotechnology 2005;25(6):65-70
The expression cDNA library of A. tabescens was constructed by SMART technique, which useλTriplEx2 as a vector. The titer and the percentage of the constructed library were about 1.0 × 106pfu/mland 98.3% respectively, and the titer and the capacity of the amplified library were about 3.1 × 108pfu/mland 4.2 × 1010. The library was used to provide expressed sequence tags (ESTs). 147 Expressed SequenceTaqs (ESTs) were gained from 176 clones, which were selected randomly and sequenced at the 5'end. Thesequences were submitted to the EMBL database. Blasting the sequences in the GenBank, 43 of them werefound that they have significant similarity with data in GenBank. EST AJ620046 was has significantsimilarity with the arabinosidase of Bacteroides thetaiotaomicron. Using SMART-RACE a full-length cDNA ofAJ620046 was successfully obtained. In order to initially characterize the biochemical properties ofAJ620046, the ORF of AJ620046 named AF was cloned and expressed in Pichia Pastoris yeast.Recombinant pHIL-S1-AF constructed by inserting AF into pHIL-S1 was transformed into Pichia PastorisGS115. Preliminary experiments indicated that AJ620046 was expressed as a 32 kDa protein in recombinantyeast.
8.A study on the relationship between alcohol drinking and fatty liver
Fang-Chao ZHU ; Chun-Wei HUANG ; Da PAN
Journal of Preventive Medicine 2015;(5):450-453
Objective To explore the relationship between alcohol drinking and fatty liver disease and its influencing factors.Methods From October to December,2013,a total of 394 people who underwent a physical examination in the medical examination center of Wenzhou central hospital were selected for this study.Anthroposomatology measurement and biochemical tests were conducted.Results There were significantly statistical differences of triglyceride,uric acid and cholesterol in different drinking groups (P <0.05).The prevalence of high triglyceride and uric acid were increased with alcohol consumption (P <0.05).There was no significant difference of alcohol consumption between fatty liver and non -fatty liver group (P =0.42).Logistic regression showed that waist -hip ratio,hypertension,overweight,obesity and hypertriglyceridemia were risk factors of fatty liver,while daily alcohol consumption cannot be thought as risk factor yet. Conclusion Waist -hip ratio,hypertension,overweight -obesity and hypertriglyceridemia were the risk factors of fatty liver.Alcohol consumption could contribute to the prevalence of triglyceride and uric acid.
9.The effect of separating red blood cells from bone marrow graft in vitro by methylcellulose.
Miao ZHANG ; Bo-Long ZHANG ; Hai-Jie JIN ; Chun-Ji GAO ; Fang-Ding LOU ; Wan-Ming DA
Journal of Experimental Hematology 2002;10(6):561-563
It has need to separate red blood cells (RBC) from marrow graft in ABO group unmatched BMT and auto-BMT with purging tumor cells, the separating effect of methylcellulose was observed. The mixture of 0.5% methylcellulose and bone marrow was laid up in an open transfusion system, and then sedimentation of RBC was performed in the transfusion tube. The separating results of 18 marrow grafts showed that the recovery rates of mononuclear cells and CD34(+) cells were (83.8 +/- 55.2)% and (90.3 +/- 7.2)%, respectively. RBC residual rate was (4.3 +/- 1.5)%. The yield of CFU-GM was (60.8 +/- 22.4)/2 x 10(5) MNC, and there was no difference to [(69.8 +/- 23.4)/2 x 10(5) MNC] yielded from same marrow samples, separated by Ficoll-Hypaque separation. It is concluded that this method could be used for bone marrow transplantation.
Bone Marrow Transplantation
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methods
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Cell Separation
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methods
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Erythrocytes
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immunology
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Humans
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Methylcellulose
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pharmacology
10.The primary study on the detection of sterigmatocystin by biologic enzyme electrode modified with the multiwall carbon nanotubes.
Dong-Sheng YAO ; Sheng-Mei WEN ; Da-Ling LIU ; Chun-Fang XIE ; Yan BAI ; Yan-Hong RAN
Chinese Journal of Biotechnology 2004;20(4):601-606
Sterigmatocystin (ST), the secondary metabolite of many kinds of filamentous fungi, is a potent carcinogen structurally related to the aflatoxins (AFT). With similar chemical structure, sterigmatocystion behaves much the homogeneous properties to aflatoxins, both of these mycotoxins exhibit similar biological properties due to their bisfuranoid structure. Since the common, and even heavier pollution, found in foods and feeds-stuff, sterigmatocystion is more harmful than aflatoxins. The reported detection methods of sterigmatocystion included the Thin-layer Chromatography, the High-Performance-Liquid Chromatography, the Enzyme-Linked Immunosorbant Assay and the PCR detection to the toxic gene, however studies about both easy and inexpensive electro-chemical methods have not been found. Our previous studies had discovered that Sterigmatocystin (ST) exist similar sensitivity towards aflatoxin-detoxifizyme (ADTZ), which we had isolated from a fungus, as aflatoxin does. In this work, the preliminary study on electrochemical analysis and determination of ST with triplet electrode enzyme-biosensor system (Ag/AgCl as the reference electrode, Pt and Au as the pair and work electrode, respectively) was carried out. Multiwall-carbon-nanotube (MWNT) had been used to increase the electron transportation on electrode. In the research, the Au electrode was modified by MWNT-immobilized ADTZ, and then the voltammertric behavior of ST was studied by means of cyclic voltammogram analysis and different pulse analysis. Autoprobe CP Research Atomic Force Microscope and TECNAI 10 Transmission Electron Microscope, had been used to detect the MWNT as well as the surface of MWNT-modified ADTZ. The voltammertric behavior of ST was studied by means of cyclic voltammogram analysis and different pulse analysis. The results show that the red-ox peak potential of ST is at the point of -600 mV, the linear detection range is from 8.32 x 10(-5) to 66.56 x 10(-5) mg/mL, the detection limit is at 8.32 x 10(-5) mg/mL, and the response time is 10 seconds. This study provided a good basic work for further research.
Biosensing Techniques
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methods
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Electrochemistry
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Microscopy, Atomic Force
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Microscopy, Electron, Transmission
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Nanotubes, Carbon
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chemistry
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Sterigmatocystin
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analysis