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Biomedical subjects

H Y Xu

Publications and source records attributed to H Y Xu.

At least 19 recordsLinked to original sources

The effect of C-type natriuretic peptide on delayed rectifier potassium currents in gastric antral circular myocytes of the guinea-pig.

C-type natriuretic peptides (CNP) play an inhibitory role in smooth muscle motility of the gastrointestinal tract, but the effect of CNP on delayed rectifier potassium currents is still unclear. This study was designed to investigate the effect of CNP on delayed rectifier potassium currents and its mechanism by using conventional whole-cell patch-clamp technique in guinea-pig gastric myocytes isolated by collagenase. CNP significantly inhibited delayed rectifier potassium currents [I(K (V))] in dose-dependent manner, and CNP inhibited the peak current elicited by depolarized step pulse to 86.1+/-1.6 % (n=7, P<0.05), 78.4+/-2.6 % (n=10, P<0.01) and 67.7+/-2.3 % (n=14, P<0.01), at concentrations of 0.01 micromol/l, 0.1 micromol/l and 1 micromol/l, respectively, at +60 mV. When the cells were preincubated with 0.1 micromol/l LY83583, a guanylate cyclase inhibitor, the 1 ?micromol/l CNP-induced inhibition of I(K (V)) was significantly impaired but when the cells were preincubated with 0.1 micromol/l zaprinast, a cGMP-sensitive phosphodiesterase inhibitor, the 0.01 micromol/l CNP-induced inhibition of I(K (V)) was significantly potentiated. 8-Br-cGMP, a membrane permeable cGMP analogue mimicked inhibitory effect of CNP on I(K (V)). CNP-induced inhibition of I(K (V)) was completely blocked by KT5823, an inhibitor of cGMP-dependent protein kinase (PKG). The results suggest that CNP inhibits the delayed rectifier potassium currents via cGMP-PKG signal pathway in the gastric antral circular myocytes of the guinea-pig.

Animals↗

Structural, optical, and magnetic properties of Mn-doped ZnO thin film.

The Zn(1-x)Mn(x)O (x = 0, 0.16, and 0.25) thin films were grown on fused quartz substrates by reactive magnetron cosputtering. X-ray-diffraction measurement revealed that all the films were single phase and had wurtzite structure with c-axis orientation. As Mn concentration increased in the Zn(1-x)Mn(x)O films, the c-axis lattice constant and band-gap energy increased gradually. In Raman-scattering studies, an additional Mn-related vibration mode appeared at 520 cm(-1). E(2H) phonon line of Zn(1-x)Mn(x)O alloy was broadened asymmetrically and redshifted as a result of microscopic structural disorder induced by Mn(2+) random substitution. The Zn(0.84)Mn(0.16)O film exhibited a ferromagnetic characteristic with a Curie temperature of approximately 62 K. However, with increasing Mn concentration to 25 at. %, ferromagnetism disappeared due to the enhanced antiferromagnetic superexchange interactions between neighboring Mn(2+) ions.

Journal Article↗

Structural analysis of anti-tumor heteropolysaccharide GFPS1b from the cultured mycelia of Grifola frondosa GF9801.

A 21-kDa heteropolysaccharide, coded as GFPS1b, was obtained from the cultured mycelia of Grifola frondosa GF9801 by hot-water extraction, ethanol precipitation, and fractioned by DEAE Sepharose Fast-flow, followed by the purification with Sephadex G-100 column chromatography using an AKTA purifier. It exhibited more potent anti-proliferative activity on MCF-7 cells than other polysaccharide fractions. GFPS1b was an acidic polysaccharide with approximately 16.60% protein and 4.3% uronic acid. Gas chromatography of absolute acid hydrolysate of GFPS1b suggested that it was composed of D-glucose, D-galactose, and L-arabinose with a molar ratio of 4:2:1. Periodate oxidation, Smith degradation, partial acid hydrolyzation, methylation analysis, FT-IR, and (1)H, (13)C NMR spectroscopy analysis revealed that GFPS1b had a backbone consisting of alpha-(1-->4)-linked D-galacopyranosyl and alpha-(1-->3)-linked D-glucopyranosyl residues substituted at O-6 with glycosyl residues composed of alpha-L-arabinose-(1-->4)-alpha-D-glucose (1--> linked residues.

Antineoplastic Agents↗

Improving the blood compatibility of polyurethane using carbon nanotubes as fillers and its implications to cardiovascular surgery.

Blood compatibility has been an occlusion for biomaterials used in the cardiovascular system. In this work, a multiwalled carbon nanotubes-polyurethane composite (MWNT-PU) was prepared through a controlled co-precipitation. The surface chemical composition of treated carbon nanotubes was analyzed with XPS and the thermal behaviors of composite were characterized by DSC. The platelet adhesion and activation caused by the composite were evaluated by using SEM and flow cytometric analysis, respectively, and the disruption of red blood cells was analyzed through measuring the absorbance of free hemoglobin. The experimental results demonstrated that: (1) Multiwalled carbon nanotubes (MWNTs) with oxygen-containing functional groups could be well dispersed in polyurethane matrix through a controlled coprecipitation; (2) the composite surface displayed a significantly improved anticoagulant function, which can be indicative of the promising potentials of carbon nanotube-based materials in the implants and medical devices applied in blood-contacting environments.

Biocompatible Materials↗

Optimization of the medium composition for production of mycelial biomass and exo-polymer by Grifola frondosa GF9801 using response surface methodology.

In this work, a three-level Box-Behnken factorial design was employed combining with response surface methodology (RSM) to optimize the medium composition for the production of the mycelial biomass and exo-polymer in submerged cultures by Grifola frondosa GF9801. A mathematical model was then developed to show the effect of each medium composition and their interactions on the production of mycelial biomass and exo-polymer. The model estimated that, a maximal yield of mycelial biomass (17.61 g/l) could be obtained when the concentrations of glucose, KH2PO4, peptone were set at 45.2 g/l, 2.97 g/l, 6.58 g/l, respectively; while a maximal exo-polymer yield (1.326 g/l) could be achieved when setting concentrations of glucose, KH2PO4, peptone at 58.6 g/l, 4.06 g/l and 3.79 g/l, respectively. These predicted values were also verified by validation experiments. Compared with the values obtained by other runs in the experimental design, the optimized medium resulted in a significant increase in the yields of mycelial biomass and exo-polymer. Maximum mycelial biomass yield of 22.50 g/l was achieved in a 15-l fermenter using the optimized medium.

Biomass↗

An autosomal genomic scan for loci linked to type 2 diabetes in northern Han Chinese.

We report the results of a genome-wide scan conducted in 219 individuals from 34 large multiplex nuclear pedigrees from the northern Han Chinese population at an average resolution of about 10 cM. Nonparametric two-point and multipoint linkage analyses were performed to detect evidence of linkage with type 2 diabetes in this study. On chromosome 1 four regions showed evidence of linkage with type 2 diabetes in northern Han Chinese. Of these regions a marker D1S193 (73 cM) showed evidence of linkage (two-point nonparametric linkage 2.409), and another region (around 190 cM) was a replication of several other studies performed in different ethnic populations. Evidences of linkage have been confirmed by typing additional markers (average distance 1-5 cM) flanking these two positive regions on chromosome 1. We also found indication of linkage with type 2 diabetes on chromosomes 2, 10, 12, 18, 20, and 22 by two-point linkage analyses.

Adult↗

Further identification and characterization of novel intermediate and mature cleavage products released from the ORF 1b region of the avian coronavirus infectious bronchitis virus 1a/1b polyprotein.

The coronavirus 3C-like proteinase is one of the viral proteinases responsible for processing of the 1a and 1a/1b polyproteins to multiple mature products. In cells infected with avian coronavirus infectious bronchitis virus (IBV), three proteins of 100, 39, and 35 kDa, respectively, were previously identified as mature cleavage products released from the 1b region of the 1a/1b polyprotein by the 3C-like proteinase. In this report, we show the identification of two more cleavage products of 68 and 58 kDa released from the same region of the polyprotein. In addition, two stable intermediate cleavage products with molecular masses of 160 and 132 kDa, respectively, were identified in IBV-infected cells. The 160-kDa protein was shown to be an intermediate cleavage product covering the 100- and 68-kDa proteins, and the 132-kDa protein to be an intermediate cleavage product covering the 58-, 39-, and 35-kDa proteins. Immunofluorescent staining of IBV-infected cells and cells expressing individual cleavage products showed that the 100-, 68-, and 58-kDa proteins were associated with the membranes of the endoplasmic reticulum, and the 39- and 35-kDa proteins displayed diffuse distribution patterns.

Animals↗

Induction of caspase-dependent apoptosis in cultured cells by the avian coronavirus infectious bronchitis virus.

Avian coronavirus infectious bronchitis virus (IBV) is the causative agent of chicken infectious bronchitis, an acute, highly contagious viral respiratory disease. Replication of IBV in Vero cells causes extensive cytopathic effects (CPE), leading to destruction of the entire monolayer and the death of infected cells. In this study, we investigated the cell death processes during acute IBV infection and the underlying mechanisms. The results show that both necrosis and apoptosis may contribute to the death of infected cells in lytic IBV infection. Caspase-dependent apoptosis, as characterized by chromosomal condensation, DNA fragmentation, caspase-3 activation, and poly(ADP-ribose) polymerase degradation, was detected in IBV-infected Vero cells. Addition of the general caspase inhibitor z-VAD-FMK to the culture media showed inhibition of the hallmarks of apoptosis and increase of the release of virus to the culture media at 16 h postinfection. However, neither the necrotic process nor the productive replication of IBV in Vero cells was severely affected by the inhibition of apoptosis. Screening of 11 IBV-encoded proteins suggested that a 58-kDa mature cleavage product could induce apoptotic changes in cells transiently expressing the protein. This study adds one more example to the growing list of animal viruses that induce apoptosis during their replication cycles.

Amino Acid Chloromethyl Ketones↗

[Pharmacokinetics of meloxicam in healthy Chinese volunteers].

AIM: To assess the pharmacokinetic profile of single doses of meloxicam in healthy Chinese volunteers. METHODS: The plasma concentrations of meloxicam after an oral dose of 15 mg to twenty healthy male volunteers were analyzed by means of a validated HPLC method. The pharmacokinetic parameters were subjected to Shapiro-Wilk test to determine whether these data were fitted to a normal distribution. RESULTS: The twenty volunteers can be classified into extensive metabolizers and poor metabolizers according to pharmacokinetic parameters. The main parameters in the two groups obtained were as follows: T 1/2 were 21 +/- 4 and 38 +/- 9 h, AUC0-infinity were 49 +/- 10 and 110 +/- 8 micrograms.h.mL-1, respectively. Even the AUC data in extensive metabolizers were 1.7 times as that reported in White volunteers following the same doses of meloxicam. CONCLUSION: There were significant individual differences in the pharmacokinetics of meloxicam in Chinese volunteers, which may be due to the genetic polymorphism of CYP2C9.

Adult↗

[Determination of bambuterol in human plasma by liquid chromatography-electrospray tandem mass spectrometry: application to pharmacokinetic study].

AIM: To develop a sensitive, specific and accurate method for quantifying bambuterol in human plasma and to study pharmacokinetics of bambuterol in male healthy Chinese. METHODS: Plasma samples were prepared based on a simple liquid-liquid extraction. The extracted samples were analyzed on liquid chromatography using a Zorbax SB C18 column interfaced with a triple quadrupole tandem mass spectrometer and detected by use of selected reaction monitoring mode. RESULTS: The linear calibration curves were obtained in the concentration range of 0.05-4.0 ng.mL-1. The limit of quantification was 0.05 ng.mL-1. The intra- and inter-run precision was measured to be below 7%. The inter-run accuracy was less than 8% for the analyte. After an oral administration of 10 mg bambuterol hydrochloride to 18 healthy Chinese volunteers the main pharmacokinetic parameters of bambuterol were as follows: Tmax was (2.3 +/- 1.3) h; Cmax was (3.95 +/- 2.20) ng.mL-1; T1/2 was (11.4 +/- 6.1) h and AUC0-t was (26.85 +/- 11.77) ng.h.mL-1. CONCLUSION: The method is shown to be accurate, robust and convenient, and suitable for pharmacokinetic studies of bambuterol. It was found that there was marked inter-individual difference in the pharmacokinetics of bambuterol in Chinese volunteers after a single oral dose, which may be attributed to the difference of activity of cholinesterase, an enzyme catalyzing bambuterol metabolism.

Adult↗

[Rapid analysis of terbutaline by combined solid phase extraction/liquid chromatography tandem mass spectrometry].

AIM: To develop a liquid chromatography-electrospray ionization tandem mass spectrometry method for rapid analysis of terbutaline at level of 50 pg.mL-1 in human plasma. METHODS: Samples containing terbutaline and salbutamol (internal standard, IS) were extracted using C18 solid-phase extraction cartridges, followed by liquid chromatographic separation and mass spectrometric detection. The mobile phase consisted of acetonitrile-water-formic acid (20:80:1), at a flow-rate of 0.4 mL.min-1. Selected reaction monitoring with mass transitions m/z 226-->151 and m/z 240-->148 were used for terbutaline and IS, respectively. RESULTS: The chromatographic analysis time for each sample was approximately 3.8 min. The assay was linear from 0.05 to 8.0 ng.mL-1. The between-run precision and accuracy of the quality controls (QCs, 0.1, 0.4 and 4.0 ng.mL-1) were characterized by relative standard deviation (RSD) of 2.5% to 7.1% and relative errors of -3.1% to 5.7%, respectively. The within-run precision of QCs was characterized by RSD of 4.2% to 6.6%. This method was applied to the analysis of samples taken up to 60 h after oral administration of 10 mg bambuterol in healthy volunteers. CONCLUSION: The method is shown to be suitable for clinical investigation of terbutaline pharmacokinetics, which offers advantages of specificity, speed, and greater sensitivity over previously reported methods.

Bronchodilator Agents↗

Developmental changes of NADPH-diaphorase positive structures in the isthmic nuclei of the chick.

Nicotine adenine dinucleotide phosphate-diaphorase staining was used to study nitric oxide synthase activity and distribution in the midbrain visual structures of white leghorn chick embryos and post-hatched chickens. Enzyme staining first appeared in the isthmic region at the tenth embryonic day (E10) in the neuropil of the nucleus isthmi, pars parvocellularis. At E11 faint enzyme positivity appeared also in the nucl. isthmi pars magnocellularis, the nucl. semilunaris and the nucl. isthmo-opticus. The staining intensity of the isthmic nuclei dramatically increased between the 12.5th and the 13th days of incubation. The nucl. isthmi, pars parvocellularis showed the strongest enzyme reaction throughout embryonic life. A day before hatching all the isthmic nuclei were heavily stained, however, nicotine adenine dinucleotide phosphate-diaphorase-positive cells occurred exclusively in the nucleus isthmo-opticus. In the tectum opticum, intensely stained cells occupied the stratum fibrosum et griseum superficiale. The layer containing the projection neurons to the isthmo-optic nucleus was unstained. In the isthmic region, the intensity of staining surpassed that of the tectum and reached its maximum at E17 and then slowly decreased till the end of the experimental period (120 days post-hatched). The tractus isthmo-opticus showed nicotine adenine dinucleotide phosphate-diaphorase activity throughout the investigated period of life of the chicken, but the tractus tectoisthmo-opticus was unstained. Our results suggest that in the isthmic nuclei, nicotine adenine dinucleotide phosphate-diaphorase-positive neurons occur only in the isthmo-optic nucleus and optic tectum. The other positively stained structures are the fibers and terminals of tectal cells. In most brain areas nicotine adenine dinucleotide phosphate-diaphorase indicates nitric oxide synthase that produces nitric oxide. The transient appearance of this molecule is probably necessary for neuronal differentiation or the establishment of synaptic connections in the isthmic nuclei, and these developmental changes are under the control of the optic tectum.

Age Factors↗

Proteolytic mapping of the coronavirus infectious bronchitis virus 1b polyprotein: evidence for the presence of four cleavage sites of the 3C-like proteinase and identification of two novel cleavage products.

We have previously reported that the 3C-like proteinase of the coronavirus infectious bronchitis virus (IBV) is responsible for processing of the 1a and 1a/1b polyproteins to three mature products of 24, 10, and 100 kDa (Liu et al., 1994, 1997; Ng and Liu, 1998). The C-terminal cleavage site of the 100-kDa protein was defined to be the Q891(1b)-S892(1b) dipeptide bond encoded by nucleotides 15,129 to 15,134 (Liu and Brown, 1995). In this report, other cleavage sites of the 3C-like proteinase in the polyprotein encoded by the ORF 1b region were mapped by coexpression, deletion, and site-directed mutagenesis studies. Using two ORF 1b-specific antisera, V58 and V17, three more Q-S(G) dipeptide bonds, encoded by nucleotides 16,929 to 16,934, 18,492 to 18,497, and 19,506 to 19,511, respectively, were demonstrated to be the cleavage sites of the 3C-like proteinase. Cleavage at these four positions would result in the release of four mature products with molecular masses of approximately 68, 58, 39, and 35 kDa. Among them, the 39- and 35-kDa proteins were specifically identified in IBV-infected cells. Taken together with the 100-kDa protein previously identified, these results suggest that the ORF 1b region of IBV mRNA1 may be able to encode five mature products.

Animals↗

Proteolytic processing of the polyprotein encoded by ORF1b of the coronavirus infectious bronchitis virus (IBV).

We present here evidence demonstrating that four previously predicted Q-S(G) cleavage sites, encoded by the IBV sequences from nucleotide 15,129 to 15,134, 16,929 to 16,934, 18,492 to 18,497, and 19,506 to 19,511, respectively, can be recognised and transcleaved by the 3C-like proteinase. Five mature products with sizes of approximately 100 kDa, 65 kDa, 63 kDa, 42 kDa and 35 kDa are released from the ORF1b polyprotein by the 3C-like proteinase-mediated cleavage at these positions. Meanwhile, expression of plasmids containing only the ORF1b region showed no autocleavage of the polyprotein encoded, suggesting that the 3C-like proteinase may be the sole proteinase involved in processing of the 1b polyprotein. These data may therefore represent a complete processing map of the polyprotein encoded by ORF1b of mRNA1.

Antibodies↗

Regulation of mRNA 1 expression by the 5'-untranslated region (5'-UTR) of the coronavirus infectious bronchitis virus (IBV).

In this report, we show that expression of the coronavirus IBV mRNA1 is regulated by its 5'-UTR. Evidence presented demonstrates that the IBV sequence from nucleotide 1 to 1904 directs very inefficient synthesis of a product of approximately 43 kDa. Deletion of either the first 362 bp or the whole part of the 5'-UTR, however, dramatically increased the expression of the 43 kDa protein species. The mechanisms involved were investigated by two different approaches. Firstly, translation of the same construct in the presence of [3H]-leucine ruled out the possibility that initiation of small reading frames from non-AUG codons located in the 5'-UTR may compete with the authentic AUG initiation codon, and therefore inhibit the expression of ORF 1a. Secondly, expression and deletion analyses of a dicistronic construct showed that translation of the 43 kDa protein was initiated by ribosome internal entry mechanism. These studies suggest that a 'weak' ribosome internal entry signal is located in the 5'-UTR and is involved in the regulation of mRNA1 expression.

5' Untranslated Regions↗