Biomedical subjects

Yuguang Du

Publications and source records attributed to Yuguang Du.

14 recordsLinked to original sources

PubMed2006

Induction of tobacco genes in response to oligochitosan.

Oligochitosan has a variety of biological activities. To understand its mechanism, DDRT-PCR, reverse Northern blot and quantitative relative RT-PCR were used to identify and isolate genes whose transcription were altered in cultured Nicotiana tabacum (var. Samsun NN) plants that were treated with oligochitosan. Three genes whose mRNA levels significantly changed in response to oligochitosan were isolated and identified. One gene is up-regulated, and two genes are down-regulated. These genes encode a DNAJ heat shock N-terminal domain-containing protein, a histone H1 gene and a hypothetical protein, whose function is unknown. The results suggest that the usefulness of mRNA differential display technique for the detection of plant metabolic pathways affected by oligochitosan.

Blotting, Northern↗

PubMed2006

Isolation and characterization of angiotensin I-converting enzyme inhibitory peptides derived from porcine hemoglobin.

Animal blood is potentially an untapped source of drugs and value-added food production. More than 400 million pigs are slaughtered each year but porcine blood is usually discarded in China. This study describes the isolation and characterization of angiotensin I-converting enzyme (ACE) inhibitory peptides derived from porcine hemoglobin. The most active hydrolysate was obtained from the peptic digestion of porcine hemoglobin. After the purification of ACE-inhibitory peptides with Sephadex LH-20 gel chromatography and reversed-phase high-performance liquid chromatography (RP-HPLC) on C(18) column, two active fractions were obtained. They were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS). They were LGFPTTKTYFPHF and VVYPWT, corresponding to the 34-46 fragment of the alpha chain and the 34-39 fragment of the beta chain of porcine hemoglobin, with IC(50) values of 4.92 and 6.02 microM, respectively. They were the first found from porcine hemoglobin; in particular, LGFPTTKTYFPHF was a novel ACE-inhibitory peptide. In addition, the purified ACE inhibitors both competitively inhibited ACE, and maintained inhibitory activity even after incubation with gastrointestinal proteases. This suggests that these peptides might have a potential antihypertensive effect.

Amino Acid Sequence↗

PubMed2006

Structural features in carrageenan that interact with a heparin-binding hematopoietic growth factor and modulate its biological activity.

The effects of carrageenans' structural features on its interaction with granulocyte colony-stimulating factor (G-CSF) and on the growth and differentiation of a G-CSF dependent leukemia cell line (NFS-60) were studied. lambda, iota, and kappa carrageenans, with decreasing contents of sulfation, bound to G-CSF with binding constants of (6.2+/-0.6) x 10(5)M(-1), (7.4+/-0.5) x 10(5)M(-1) and (6.0+/-0.4) x 10(5)M(-1), and with 27.7+/-0.2, 17.4+/-0.1 and 8.4+/-0.1 binding sites, respectively. However, kappa carrageenan oligosaccharide had no affinity for G-CSF. The three carrageenans significantly inhibited G-CSF-induced growth of NFS-60 cells. The high sulfate content lambda carrageenan could also induce the maturation of the cells, but relatively low sulfate content iota and kappa carrageenans could not. The results suggested that G-CSF-carrageenan bindings were dependent on carrageenans' sulfate contents and chain lengths, which could also affect the growth and differentiation of NFS-60 cells.

Carbohydrate Conformation↗

PubMed2006

A normal mucin-binding lectin from the sponge Craniella australiensis.

A lectin, Craniella australiensis (CAL), was isolated from sponge C. australiensis by ion-exchange on DEAE-Sephacel and purified by gel filtration on Sephadex G-150 and HPLC on DEAE-5PW. The purified lectin was a trimeric protein as revealed by SDS-PAGE and MALDI-TOF analysis. SDS-PAGE showed that the CAL protein had a molecular mass of 54 kDa, and consisted of three 18 kDa subunits. Gel filtration of purified lectin on Sephadex G-200 indicates that it exists as a 54 kDa protein in its native state. The amino acid composition was rich in Thr and Glx. CAL was found to agglutinate native and trypsinized human A, B erythrocytes, and agglutinate native erythrocytes of mouse, sheep, rabbit and chicken, and trypsinized erythrocytes of sheep and rabbit. The hemagglutination activity was inhibited by glycoproteins such as PSM and asialo-PSM, but not by any of the monosaccharides tested. The activity was stable between 20 and 70 degrees C. Significant CAL activity was observed between pH 5 and 8. The lectin reaction is independent of the presence of divalent cations Ca2+ and Mg2+. The sequence of N-terminal residues of CAL was determined as TSSCQSIVVE. The lectin showed a potent mitogenic response towards BALB/c splenocytes.

Amino Acid Sequence↗

PubMed2006

Interactions of dextran sulfates with granulocyte colony-stimulating factor and their effects on leukemia cells.

The interactions between granulocyte colony-stimulating factor (G-CSF) and dextran sulfate (DS) with different chain lengths and sulfate contents were studied by capillary zone electrophoresis. It was found that DS with a molecular mass of 500 kDa (DS500) could bind to G-CSF and the binding constant and binding sites were determined using Scatchard plot to be 1.17 x 10(6) M(-1) and 3, respectively. DS with a molecular mass of 40 kDa also had the affinity to G-CSF and the binding constant and binding sites were 1.01 x 10(6) M(-1) and 8, respectively. However, DS with a molecular mass of 8 kDa and the non-sulfated saccharide, dextran, had no affinity to G-CSF. The results indicate that the interactions between G-CSF and DS are dependent on the chain lengths and sulfate contents of the saccharides. In addition, the effects of G-CSF-binding DS on a G-CSF-dependent leukemia cell line were investigated using biological methods. Results show that DS500 plus G-CSF has potential therapeutic effect on cancers because their combination could inhibit the growth and induce the differentiation of the leukemia cells.

Animals↗

PubMed2006

Quantitative investigation of the interaction between granulocyte-macrophage colony-stimulating factor and heparin by capillary zone electrophoresis.

The interactions between granulocyte-macrophage colony-stimulating factor (GM-CSF) and heparin or low-molecular weight heparin (LMWH) were studied by CZE. It was found that GM-CSF could bind to both heparin and LMWH. The binding constants were calculated from Scatchard regression to be (6.5 +/- 0.8) x 10(5)/M and (11.2 +/- 0.7) x 10(5)/M, respectively. The specificity of the interaction between GM-CSF and heparin was also studied by employing another sulfated K carrageenan oligosaccharide as a control. Results showed that K carrageenan oligosaccharide could not interact with GM-CSF, indicating that GM-CSF could specifically interact with heparin.

Carrageenan↗

PubMed2005

Pharmacological studies of anti-diarrhoeal activity of Gentianopsis paludosa.

The pharmacological studies of anti-diarrhoeal activity of Gentianopsis paludosa were carried out. The 75% ethanol extract (100-400 mg/kg) of Gentianopsis paludosa inhibited castor oil-induced diarrhoea and also reduced gastrointestinal motility in the charcoal meal test in mice. In the rabbit-isolated ileum, the extract showed inhibitory effects not only on its spontaneous contraction, but also on acetylcholine (Ach, 0.1 mM)- and KCl (60 mM)-induced contractions. Furthermore, the ethanol extract (0.4-0.6 mg/ml) shifted the CaCl2 dose-effect curves to the right in a non-competitive manner and significantly reduced the acetylcholine-induced contraction of isolated ileum in Ca2+-free solution like verapamil (0.1 mM), which suggest that the extract interferes with Ca2+ movement. These results support the traditional use of Gentianopsis paludosa in the treatment of diarrhoea.

Acetylcholine↗

PubMed2005

Capillary zone electrophoresis characterization of low molecular weight heparin binding to interleukin 2.

A method based on capillary zone electrophoresis (CZE) was used to study the interaction between low molecular weight heparin (LMWH) and interleukin 2 (IL-2). The results showed that the increase of the concentration of LMWH led to the decrease of the peak height and the increase of the peak width of IL-2, but the peak areas were kept constant. The binding constant of IL-2 with LMWH was calculated as 1.2 x 10(6)M(-1) by Scatchard analysis, which is in good agreement with the results found in the references using enzyme-linked immunosorbent assay (ELISA). The results demonstrated that the interaction between IL-2 and LMWH is of fast on-and-off kinetic binding reaction. CZE might be used to study not only slow on-and-off rates interactions, but also fast on-and-off rates ones. The binding constant can be calculated easily, and the method can be applied to study a wide range of heparin-protein interactions.

Benzoic Acid↗

PubMed2005

Separation, identification, and interaction of heparin oligosaccharides with granulocyte-colony stimulating factor using capillary electrophoresis and mass spectrometry.

A capillary electrophoresis (CE) method was developed for the separation of heparin oligosaccharides compatible to study the interactions between the oligosaccharides and granulocyte-colony stimulating factor (G-CSF). Unfractionated heparin was eliminitively degraded to heparin oligosaccharides by an endolytic heparinase. The degraded smaller oligosaccharides (M(r) < 1000) were baseline-separated by CE under a 50 mM phosphate buffer (pH 9.0) in 10 min. Standard heparin disaccharides and larger oligosaccharides (1000 < M(r) < 8000) were all separated under optimized separation conditions. Compared with standard heparin disaccharides, smaller oligosaccharides contained one nonsulfated, two monosulfated, and two disulfated disaccharides, but trisulfated disaccharides were not found. The smaller oligosaccharides were also identified and molecular mass was deduced by electrospray ionization-mass spectrometry (ESI-MS). Furthermore, interactions between G-CSF and the oligosaccharides were studied by using capillary zone electrophoresis (CZE) under the above separation conditions. It was found that larger oligosaccharides could interact with G-CSF while smaller oligosaccharides were not observed to bind to G-CSF under the experimental conditions. In conclusion, the purified heparinase could selectively degrade heparin into oligosaccharides and the interaction between G-CSF and heparin was correlated with the chain length of heparin.

Chromatography, Gel↗

PubMed2005

Production and properties of an inhibitor of the Pseudomonas autoinducer by Pseudomonas aeruginosa.

An inhibitor was found in the culture fluid of Pseudomonas aeruginosa PAO1, which could inhibit the activity of the Pseudomonas autoinducer (PAI). The maximal inhibitory activity occurred in stationary phase culture sup ernatant. The PAI inhibitor did not influence the cell growth and the PAI production by P. aeruginosa PAO1 when the PAI inhibitor was added into culture medium. The induced expression of lacZ in the reporter strain Agrobacterium tumefaciens NT1 was suppressed by this PAI inhibitor, whereas inhibition could be relieved by increasing the auto inducer concentration. The quorum sensing of P. aeruginosa was inhibited presumably by inhibiting the inducing activity of Pseudomonas autoinducer but not by inhibiting the production of Pseudomonas autoinducer. It was demonstrated that the structure of the PAI inhibitor was different from that of acyl-homoserine lactones.

4-Butyrolactone↗

PubMed2004

Integrated lectin affinity microfluidic chip for glycoform separation.

Lectin affinity chromatography was miniaturized into a microfluidic format, which results in improvement of performance, as compared to the conventional method. A lectin affinity monolith column was prepared in the microchannel of a microfluidic chip. The porous monolith was fabricated by UV-initiated polymerization of ethylene dimethacrylate (EDMA) and glycidyl methacrylate (GMA) in the presence of porogeneities, followed by immobilization of pisum sativum agglutinin (PSA) on the monolith matrix. Using electroosmosis as the driven force, lectin affinity chromatographies of three kinds of glycoprotein, turkey ovalbumin (TO), chicken ovalbumin (CO), and ovomucoid (OM), were carried out on the microfluidic system. All the glycoproteins were successfully separated into several fractions with different affinities toward the immobilized PSA. The integrated system reduces the time required for the lectin affinity chromatography reaction to approximately 3%, thus, the overall analysis time from 4 h to 400 s. Only 300 pg of glycoprotein is required for the whole separation process. Moreover, troublesome operations for lectin affinity chromatography are simplified.

Adsorption↗

PubMed2004

Capillary zone electrophoresis investigation of the interaction between heparin and granulocyte-colony stimulating factor.

The interaction between standard heparin, low-molecular-weight heparin (LMWH), and granulocyte-colony stimulating factor (G-CSF) was studied by capillary zone electrophoresis. Both qualitative and quantitative characterizations of the heparin-protein binding were determined. The binding constants of the two different groups of heparins with G-CSF, calculated from the Scatchard plot by regression, were 4.805 x 10(5) M(-1) and 4.579 x 10(5) M(-1), respectively. The two binding constants measured are of the same order of magnitude at 10(5) M(-1), indicating that LMWH contains most of the functional groups bound to G-CSF by standard heparin.

Electrophoresis, Capillary↗

PubMed2004

Determination of SARS-coronavirus by a microfluidic chip system.

We have developed a new experimental system based on a microfluidic chip to determine severe acute respiratory syndrome coronavirus (SARS-CoV). The system includes a laser-induced fluorescence microfluidic chip analyzer, a glass microchip for both polymerase chain reaction (PCR) and capillary electrophoresis, a chip thermal cycler based on dual Peltier thermoelectric elements, a reverse transcription-polymerase chain reaction (RT-PCR) SARS diagnostic kit, and a DNA electrophoretic sizing kit. The system allows efficient cDNA amplification of SARS-CoV followed by electrophoretic sizing and detection on the same chip. To enhance the reliability of RT-PCR on SARS-CoV detection, duplex PCR was developed on the microchip. The assay was carried out on a home-made microfluidic chip system. The positive and the negative control were cDNA fragments of SARS-CoV and parainfluenza virus, respectively. The test results showed that 17 positive samples were obtained among 18 samples of nasopharyngeal swabs from clinically diagnosed SARS patients. However, 12 positive results from the same 18 samples were obtained by the conventional RT-PCR with agarose gel electrophoresis detection. The SARS virus species can be analyzed with high positive rate and rapidity on the microfluidic chip system.

Autopsy↗

PubMed2003

Analysis of chicken and turkey ovalbumins by microchip electrophoresis combined with exoglycosidase digestion.

The polypeptide and carbohydrate patterns of two glycoproteins, chicken ovalbumin (CO) and turkey ovalbumin (TO), were analyzed by microchip electrophoresis (ME), following digestion with proteases and exoglycosidases. Glycopeptides derived from ovalbumin were obtained by digestion with Pronase, followed by dialysis, and then separated by ME. Using CO as model, the method was developed to deduce the structure of glycans from glycoproteins by comparing the electropherograms of glycopeptides with and without digestion of exolycosidases. Applying the same approach, the structure of oligosaccharides linked to TO was determined. TO was found to contain high-mannose type oligosaccharides and oligosaccharides with terminal N-acetylglucosamine residues. The complete primary analysis of CO and TO by ME described in this paper provides a basis for an analysis of glycoproteins with an integrated microfluidic chip.

Amino Acid Sequence↗