1.Formation of pyrrole adducts in 2,5-hexanedione-containing human serum cultured in vitro.
Ming-xing ZHU ; Hong-yin YIN ; Ke-qin XIE
Chinese Journal of Industrial Hygiene and Occupational Diseases 2013;31(8):611-614
OBJECTIVETo investigate the relationship between formation of pyrrole adducts and concentration of 2, 5-hexanedione (2, 5-HD) and to provide an experimental basis for the study on toxicity of n-hexane.
METHODSSerum samples were collected from normal persons and were then filtered and sterilized. They were mixed with 2,5-HD to obtain sera with final 2, 5-HD concentrations of 10, 25, 50, 100, and 200 mg/L, and blank serum was also prepared. The sera were cultured at 37°C and taken at different time points. Colorimetry was used to quantify the pyrrole adducts formed in sera, and gas chromatography was used to measure the remaining 2, 5-HD levels in sera.
RESULTSThe content of pyrrole adducts increased as the culture proceeded and was dependent on the dose of 2, 5-HD; at the end of the experiment, the content of pyrrole adducts differed significantly across all concentration groups (P < 0.5). The concentrations of 2,5-HD decreased as the culture proceeded; at the end of the experiment, the concentrations of 2, 5-HD, from the highest to the lowest, decreased by 29%, 55%, 22%, 44%, and 40%, respectively. The decrease in 2, 5-HD had a positive correlation with the increase in pyrrole adducts, and the correlation coefficients for 200∼10 mg/L 2, 5-HD were 0.865, 0.697, 0.835, 0.823, and 0.814, respectively.
CONCLUSIONThe content of formed pyrrole adducts increases as the concentration of 2,5-HD rises; there is a positive correlation between the decrease in 2, 5-HD and the increase in pyrrole adducts in human serum.
Hexanones ; chemistry ; Humans ; Oxidation-Reduction ; Pyrroles ; chemistry ; Serum ; chemistry
3.Method for determining 2, 4-D butylate in serum by gas chromatography.
Baoying JIN ; Yi QIAN ; Shuming DU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2014;32(2):145-146
OBJECTIVETo establish a method for determining 2, 4-D butylate in serum by gas chromatography (GC)and to provide a basis for the diagnosis and treatment of clinical poisoning.
METHODSSerum 2, 4-D butylate level was determined by the following steps: mixing serum (0.5 ml)with trichloromethane (2.0 ml), adequately shaking for extraction, standing for 5 min, centrifuging at 4 000 rpm for 10 min, blow-drying the trichloromethane layer with nitrogen, adding ethanol (50 µl)to a certain volume, adding the sample (1.0 µl), and performing GC with a hydrogen flame ionization detector.
RESULTSSerum 2, 4-D butylate level showed a linear relationship within 5∼40 µg/ml, with a regression equation of y = 1 831.6.4x-899.4 (r = 0.999 2); the minimum detectable concentration was 1.0 µg/ml. The recovery rate was 88.7%∼103.0% (relative standard deviation (RSD) 3.8%∼5.0%). The intra-day RSD and inter-day RSD were 3.87-4.92% and 3.33%∼5.34%, respectively.
CONCLUSIONThis determination method is simple, efficient, and accurate and provides a good means for rapid diagnosis and treatment of 2, 4-D butylate poisoning.
Chromatography, Gas ; methods ; Humans ; Serum ; chemistry ; Thiocarbamates ; blood
5.The molecular mechanism between baicalin metal complexes and bovin serum album.
Ming GUO ; Xian TAN ; Ying WANG ; Xiao-yan GAO ; Zhou-ling WU ; Li-jun ZHANG
Acta Pharmaceutica Sinica 2015;50(5):613-620
In this paper, the new carbon nanotube modified glassy carbon electrode (F-CNTs/GCE) was prepared to establish a new method for studying the molecular interaction mechanism between baicalin metal complexes (BMC) and bovine serum album (BSA), and the principle of this method was discussed deeply. Under the physiological condition, the thermodynamics and kinetics properties of interaction between BMC and BSA were studied by cyclic voltammetry (CV) to inference their molecular effective mechanism. The results show that the presence of F-CNTs can accelerate the electron transfer, and better response signal was showed in the BMC/BMC-BSA system. The detection of interaction of BMC-BSA used new method show that BMC-BSA generates stable thermodynamically non-covalent compounds, and the obtained average binding sites of BMC-BSA were 1.7; the number of electron transfer in BMC/BMC-BSA reaction process was 2, and non electroactive supramolecular compounds of BMC-BSA were generated by this interacting reaction. The relevant research work provides a new way to study the molecular mechanism for the interaction of drugs with protein, and with a certain reference value for discussion on the non covalent interactions.
Animals
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Cattle
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Coordination Complexes
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chemistry
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Electrodes
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Flavonoids
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chemistry
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Kinetics
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Nanotubes, Carbon
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Serum Albumin, Bovine
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chemistry
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Thermodynamics
6.Preparation and characteristics of iron oxide nanoparticles modified by bovine serum albumin.
Peng ZHAO ; Jie WENG ; Peng HOU
Journal of Biomedical Engineering 2009;26(5):1005-1009
Ultrafine iron oxide nanoparticles were prepared in aqueous solution using chemical coprecipitation method. Adsorption of bovine serum albumin (BSA) on magnetic particles was studied in the presence of silane coupling KH550. Characterizations of magnetic particles were carried out by X-ray diffraction, Transmission electron microscopy, Fourier transform infrared spectrometer and vibrating-sample magnetometer. The experimental results showed that magnetic nanoparticles were well dispersed with a small decrease of the magnetic saturation after modification. Magnetic nanoparticles wrapped in KH550 were favorable to the adsorption of BSA while the capability of hydrophile increased consequently, which could be used for target carrier in biomedicine.
Animals
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Cattle
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Ferrosoferric Oxide
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chemistry
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Magnetics
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Metal Nanoparticles
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chemistry
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Serum Albumin, Bovine
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chemistry
7.MC3T3-E1 osteoblasts adhesion to micropatterned surfaces.
Peiqing YING ; Gang JIN ; Zulai TAO
Journal of Biomedical Engineering 2002;19(3):370-373
Cell adhesion to material surface plays an important role in regulating cell function such as proliferation and differentiation. Surface patterning provides a useful method to control cell spatial distribution and adhesion to substance. Here microcontact printing and microfluidic channels were introduced to pattern MC3T3 E1 osteoblasts on silicon substance. Dichlordimethylsilane (DMS) was used in microcontact printing to generate the alternating domains of DMS and non-DMS, and cells preferentially adhered to the non-DMS and hydrophilic region. On the patterned surfaces generated from collagen and albumin solutions with microfluidic channels, cells preferentially localized in the collagen-coated region. The results also showed that micropatterning could be a useful method to study the effect of surface chemistry on cell adhesion and other functions.
Cell Adhesion
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Cells, Cultured
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Collagen
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chemistry
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Dimethylpolysiloxanes
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chemistry
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Osteoblasts
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physiology
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Serum Albumin, Bovine
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chemistry
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Surface Properties
8.Spectroscopic studies on binding of beta-elemene to human serum albumin.
Miao ZHANG ; Lu-Yong ZHANG ; Xian-Zhe DONG ; Ping LIU
China Journal of Chinese Materia Medica 2014;39(11):2117-2120
Beta-Elemene is an antitumor drug which is isolated from the traditional Chinese medicinal herb Curcumae Phaeocaulis Rhizoma, it is the main component of elemene which is extracted from the plant and delivered via blood circulation after intravenous injection. The antitumor effect of beta-elemene in vitro and in vivo was definite, and beta-elemene could improve the patient immunity and no sever side effect, drug resistance or bone marrow suppression were found during the clinical studies. And human serum albumin (HSA) is a primary extracellular protein which has a high concentration distribution in blood plasma and has many characteristic physiological functions. Therefore, the binding of beta-elemene to protein may be very important for absorption, distribution, metabolism and elimination. Therefore, the study on the interaction of beta-elemene with drug-carrying protein is very important. In this work, molecular binding of beta-elemene to human serum albumin (HSA) was investigated by using spectrofluorometer. the binding constants suggested that a strong interaction and the formation of a complex between beta-elemene and HSA. This clearly implies that beta-elemene can be stored and removed by the proteins in the body. Furthermore, the fluorescence quenching results showed that the HSA fluorescence was quenched by beta-elemene through static quenching mechanism. Thermodynamic parameters showed that hydrophobic interactions play a role in the binding of beta-elemene to HSA. The negative deltaH(0) and positive deltaS(0) in case of beta-elemene therefore showed that electrostatic attraction play a role in the binding of beta-elemene to HSA.
Drugs, Chinese Herbal
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chemistry
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Humans
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Kinetics
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Protein Binding
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Serum Albumin
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chemistry
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Sesquiterpenes
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chemistry
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Spectrometry, Fluorescence
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Thermodynamics
9.Spectroscopic study on interaction between cistanoside F and bovine serum albumin.
Aizhi WU ; Chaozhan LIN ; Xiaoning ZHAO ; Jialin ZHUO ; Chenchen ZHU
China Journal of Chinese Materia Medica 2012;37(10):1392-1398
OBJECTIVETo study the conjugation reaction characteristics of caffeic acid micromolecule cistanoside F and bovine serum albumin.
METHODThe interaction between bovine serum albumin (BSA) and cistanoside F that was separated from Callicarpa plant for the first time and abbreviated CF was detected by fluorescence (FS), UV-vis absorbance and circular dichroism (CD) under simulative physiological conditions.
RESULTCF-BSA's static apparent binding constant (K(a)), number of binding sites (n), efficiency of energy transfer (E), spatial distance (r), thermodynamic parameters deltaG, deltaH and deltaS and changes in alpha-helical structure content in BSA before and after CF's effect were calculated to define the binding site of CF in BSA and analyze the impact of several common metal ions on the interaction of CF and BSA.
CONCLUSIONGround state compounds formed by CF and BSA could cause intrinsic fluorescence quenching. Their binding constant K(a) of cistanoside F with BSA was 4.36 x 10(4) L x mol at 25 degrees C, the number of binding site n was 1, and the spatial distance r was 3.09 nm. The results indicated that the hydrogen bond played a major role in cistanoside F-BSA association. The displacement experiments confirmed that cistanoside F can bind to site I of BSA. In addition, the binding constant of cistanoside F with BSA was enhanced after the addition of some common metal ions Mg2+, Fe3+, Cu2+ and Zn2+. The intrinsic fluorescence of BSA was quenched by cistanoside F via forming cistanoside F-BSA complex and non-radiation energy transfer. CD spectra showed that the binding of cistanoside F with BSA induced conformational changes in BSA.
Animals ; Caffeic Acids ; chemistry ; Catechols ; chemistry ; Cattle ; Circular Dichroism ; Glycosides ; chemistry ; Serum Albumin, Bovine ; chemistry ; Spectrometry, Fluorescence ; Spectrophotometry, Ultraviolet ; Thermodynamics
10.Study on adsorption of methylene blue by sulfonated polyethersulfone II. The adsorption of methylene blue by sulfonated polyethersulfone in plasma.
Meng TIAN ; Xiaoqing SUN ; Rui ZHONG ; Xiaohua HUANG ; Fang HUANG ; Shudong SUN ; Yilun YUE
Journal of Biomedical Engineering 2008;25(1):135-138
The evaluation of the adsorption of methylene blue (MB) in plasma by sulfonated polyethersulfone (SPES) adsorbent column was carried out in this study. The results indicated the adsorption of MB by SPES adsorbent column was more efficient than that by polyethersulfone (PES). In addition, the changes of the concentration of BSA solution passing through adsorbent column along with the time and the biochemical indices of plasma before and after adsorption treatment were also investigated. The results showed that the adsorption amount of BSA by PES adsorbent column was larger than that by SPES, and the biochemical parameters such as total protein, albumin, glucose, triglyceride and total cholesterol in plasma varied slightly before and after passing through the column, which were still within the clinical indices.
Adsorption
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Humans
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Methylene Blue
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isolation & purification
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Plasma
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chemistry
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Polymers
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chemical synthesis
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chemistry
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Serum Albumin, Bovine
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chemistry
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Sulfones
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chemical synthesis
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chemistry