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Y Xia

Publications and source records attributed to Y Xia.

At least 55 records · Page 3Linked to original sources
PubMed2001

Heat-shock protein 90 augments neuronal nitric oxide synthase activity by enhancing Ca2+/calmodulin binding.

Heat-shock protein 90 (hsp90) has been shown to facilitate neuronal NO synthase (nNOS, type 1) activity in vivo. But the direct effect of hsp90 on purified nNOS has not been determined yet. Moreover, the mechanism underlying the action of hsp90 is not known. nNOS activity is primarily initiated and regulated by the binding of Ca(2+)/calmodulin (CaM). Therefore, we explored whether hsp90 modulates nNOS activity by affecting CaM binding. Recombinant rat nNOS was purified from the stably transfected cells by affinity chromatography. hsp90 increased nNOS activity in a dose-dependent manner with an EC(50) of 24.1+/-6.4 nM. In the presence of hsp90, the CaM-nNOS dose-response curve was shifted markedly to the left and the maximal activity was also elevated. Further in vitro protein-binding experiments confirmed that hsp90 increased the binding of CaM to nNOS. Taken together, these data indicate that hsp90 directly augments nNOS catalytic function and that this effect is, at least partially, mediated by CaM-binding enhancement.

Animals↗
PubMed2001

Detection of hydroxyl radicals by D-phenylalanine hydroxylation: a specific assay for hydroxyl radical generation in biological systems.

Hydroxylation of l-phenylalanine (Phe) by hydroxyl radical (*OH) yields 4-, 3-, and 2-hydroxyl-Phe (para-, meta-, and ortho-tyrosine, respectively). Phe derivative measurements have been employed to detect *OH formation in cells and tissues, however, the specificity of this assay is limited since Phe derivatives also arise from intracellular Phe hydroxylase. d-Phe, the d-type enantiomer, is not hydroxylated by Phe hydroxylase. We evaluate whether d-Phe reacts with *OH as well as l-Phe, providing a more reliable probe for *OH generation in biological systems. With *OH generated by a Fenton reaction or xanthine oxidase, d- and l-Phe equally gave rise to p, m, o-tyr and this could be prevented by *OH scavengers. Resting human neutrophils (PMNs) markedly converted l-Phe to p-tyr, through non-oxidant-mediated reactions, whereas d-Phe was unaffected. In contrast, when PMNs were stimulated in the presence of redox cycling iron the *OH formed resulted in more significant rise of p-tyr from d-Phe (9.4-fold) than l-Phe (3.6-fold) due to the significant background formation of p-tyr from l-Phe. Together, these data indicated that d- and l-Phe were equally hydroxylated by *OH. Using d-Phe instead of l-Phe can eliminate the formation of Phe derivatives from Phe hydroxylase and achieve more specific, sensitive measurement of *OH in biological systems.

Cell Survival↗
PubMed2001

Oxygen radical-mediated reduction in basal and agonist-evoked NO release in isolated rat heart.

Oxygen free radicals (OFR) play a primary role in ischemia-reperfusion-mediated vascular dysfunction and this is paralleled by a loss of endothelial nitric oxide synthase (eNOS) activity. The authors tested whether a direct exposure to OFR may affect vascular relaxation by altering nitric oxide (NO) release. Effects of electrolysis(EL)-generated OFR on basal and agonist-evoked NO release were monitored in isolated rat hearts by oxyhemoglobin assay. Electrolysis-induced changes were compared with those obtained after 30 min perfusion with NOS and cyclooxygenase (COX) inhibitors NG-nitro-L-arginine methyl ester (L-NAME, 100 microM) and indomethacin (INDO, 1 m M). Electrolysis-generated hydroxyl radical (.OH) formed by.O2-and H2O2 via the Fenton reaction as revealed by Electron Paramagnetic Resonance (EPR). After EL, basal NO release declined by 60% and coronary perfusion pressure (CPP) increased by approximately 70%. L-NAME/INDO perfusion similarly lowered NO release (-63%) but increased CPP less than EL (56+/-3%P<0.03 v post-EL). In presence of excess substrates and cofactors eNOS activity was not affected by EL. Both acetylcholine (ACh; 1 microM) and bradykinin (BK; 10 n M) had minimal effect in reversing EL-induced vasoconstriction, whereas both partially reversed L -NAME/INDO-mediated constriction. Sodium nitroprusside (SNP, 1 microM) completely reversed L-NAME/INDO constriction and partly countered that after EL (-38+/-2.5, P<0.001). Acetylcholine-evoked NO release was nearly abolished by both treatments whereas BK still elicited partial NO release after eNOS/cyclooxygenase inhibition (P<0.001) but not after EL. In conclusion, OFR severely impair NO-mediated coronary vasorelaxation affecting both basal and agonist-evoked NO release but not eNOS activity. However, EL also significantly blunts NOS/COX-independent vasodilation suggesting alteration of other vasodilatative pathways.

Acetylcholine↗
PubMed2001

Sodium-hydrogen exchangers and sodium-bicarbonate co-transporters: ontogeny of protein expression in the rat brain.

We used western blotting to examine the developmental profiles (at embryonic day 16 and postnatal days 1, 13, 23, 33 and 105) of protein expression for three sodium-hydrogen exchanger isoforms (1, 2 and 4) and for a sodium-bicarbonate co-transporter in three CNS regions (cortex, cerebellum and brainstem-diencephalon). In microsomal preparations, sodium-hydrogen exchanger isoform 1 and sodium-bicarbonate co-transporter protein expression in the CNS increases gradually from embryonic day 16 (25-40% of the adult level) to postnatal day 105. In contrast, sodium-hydrogen exchanger isoform 2 and 4 expression reaches a maximum (three to 20 times the adult level) at around three to four weeks of age. There is significant regional heterogeneity in the expression of sodium-hydrogen exchanger and sodium-bicarbonate co-transporter proteins in the rat CNS. Sodium-hydrogen exchanger isoform 1 was highly expressed in the brainstem-diencephalon, whereas the sodium-bicarbonate co-transporter was robustly expressed in the cerebellum and brainstem-diencephalon. These data indicate that the expression of sodium-hydrogen exchanger and sodium-bicarbonate co-transporter proteins varies as a function of both development and specific brain region.

Acid-Base Equilibrium↗
PubMed2001

High spatial resolution in vivo 2D (1)H magnetic resonance spectroscopic imaging of human muscles with a band-selective technique.

This report demonstrates a 2D (1)H magnetic resonance spectroscopic imaging (MRSI) technique that can address some technical difficulties often encountered in MRS studies of human muscles. A preliminary application of this whole-slice technique in human skeletal muscles demonstrates clearly noticeable differences in (1)H metabolite spectra between different human muscles. This observation illustrates the importance of multi-voxel and high spatial resolution in a heterogeneous environment. This technique is robust, can be easily implemented on a commercial MR scanner, and should prove useful for investigators in both basic and clinical (1)H MRS studies.

Adult↗
PubMed2001

3D H(aro)-NOESY-CH3NH and C(aro)-NOESY-CH3NH experiments for double labeled proteins.

Precision in the determination of the 3D structures of proteins by NMR depends on obtaining an adequate number of NOE restraints. Ambiguity in the assignment of NOE cross peaks between aromatic and other protons is an impediment to high quality structure determination. Two pulse sequences, 3D H(aro)-NOESY-CH3NH and 3D C(aro)-NOESY-CH3NH, based on a modification of a technique for simultaneous detection of 13C-1H (of CH3) and 15N-1H correlations in one measurement, are proposed in the present work. These 3D experiments, which are optimized for resolution in the 13C and 15N dimensions, provide NOE information between aromatic protons and methyl or amide protons. CH2 moieties are filtered out and the CH groups in aromatic rings are selected, allowing their NOE cross peaks to be unambiguously assigned. Unambiguous NOEs connecting aromatic and methyl or amide protons will provide important restraints for protein structure calculations.

Carbon Isotopes↗
PubMed2001

Quantification of the graphical details of collagen fibrils in transmission electron micrographs.

A novel 2D image analysis technique is demonstrated. Using the digitized images of articular cartilage from transmission electron microscopy (TEM), this technique performs a localized 'vector' analysis at each region that is large enough to include several or tens of collagen fibrils but small enough to provide a fine resolution for the whole tissue. For each small and localized region, the morphology of the collagen fibrils can be characterized by three quantities essential to the nature of the tissue: the concentration of the fibrils, the overall orientation of the fibrils, and the anisotropy of the fibrils. This technique is capable of providing new insight to the existing technology by assigning quantitative attributes to the qualitative graphics. The assigned quantities are sensitive to the fine structure of the collagen matrix and meaningful in the architectural nature of the collagen matrix. These quantities could provide a critical linkage between the ultrastructure of the tissue and the macroscopic behaviours of the material. In addition, coarse-graining the microscopic resolution of EM without compromising the essential features of the tissue's structure provides a direct view of the tissue's morphology and permits direct correlations and comparisons among interdisciplinary techniques.

Animals↗
PubMed2001

Quantitative in situ correlation between microscopic MRI and polarized light microscopy studies of articular cartilage.

OBJECTIVE: To establish the correlation between the non-invasive imaging by magnetic resonance microscopy (microMRI) and the histological imaging by polarized light microscopy (PLM) accurately, quantitatively, at the highest possible MRI resolution (13.7 microm), and based on the same piece of tissue (articular cartilage from canine shoulder joint). DESIGN: In microMRI experiments, the laminar appearance (the magic angle effect) of the proton intensity images and the anisotropic characteristics of the T(2)relaxation images were analysed. In PLM experiments, the images of the optical retardation and collagen-fibre orientation in cartilage were constructed in two dimensions. RESULTS: The T(2)profile has a distinctly asymmetric bell-shaped curve and three featured zones. The retardation profile has a non-zero minimum at the middle of the transitional zone of the tissue. The angle profile has a smooth variation across the transitional zone. These facts suggest that the collagen fibres in the transitional zone are not entirely random but have a residual order. In addition, the peak of the T(2)profile coincides with the minimum of the retardation profile, both represent the most isotropic region of the tissue. A hyperbolic tangent function was found to best describe the transition of the collagen fibres in cartilage. A set of criteria was developed for each technique to define the features in the quantitative measurements. CONCLUSIONS: The criteria offer, for the first time, a set of quantitative and objective means to subdivide the tissue thickness into the zones in histology and in MRI. It is shown that the microMRI zones based on the T(2)characteristics are statistically equivalent to the histological zones based on the collagen fibre orientation (t-probabilities of 0.730, 0.973, 0.647, 0.850 for the superficial, transitional, radial zones and the total thickness).

Animals↗
PubMed2001

Biochemical (and functional) imaging of articular cartilage.

Over the coming decades nondestructive biochemical imaging by magnetic resonance imaging (MRI) will provide an adjunct or surrogate for the destructive histologic and biochemical assays used today. A number of MRI methods demonstrate image contrast that, although influenced by the biochemical composition, is not normally specific to a particular measure of the biochemical state. The most widely used of these is T2-weighted imaging, which variably reveals collagen ultrastructure, hydration (or collagen content), and, to a lesser extent, glycosaminoglycan (GAG) concentration (each of these biochemical metrics is an important determinant of the functional integrity of cartilage). The lack of specificity of this technique (and others discussed herein) confounds efforts to improve strategies for evaluating cartilage. However, three methods permit a very specific measure of the cartilage biochemical state. Each of these three methods, explored in detail in this article, is rooted in a biophysical theory that relates the image signal intensity to a specific biochemical feature. Proton-density imaging directly measures water content (hydration), a parameter that might increase approximately 5% with significant degeneration. Magic-angle imaging, in which the angle dependence of T2 is measured, can provide a specific measure of collagen (or macromolecular) ultrastructure. The difficulty in getting the angle dependence presently precludes its use clinically. Delayed gadolinium-enhanced MRI of cartilage provides a specific measure of the distribution of GAGs. This method measures the distribution of a charged contrast agent, which in turn reflects the distribution of charge associated with GAG. This technique can be used in a clinical setting, and ongoing studies will explore its utility in monitoring therapeutic efficacy and disease progression. Although none of these techniques are presently in routine clinical use, emerging data provide promise that the future will see patient-specific biochemical analysis of cartilage, an outcome almost unimaginable 20 years ago.

Biomechanical Phenomena↗
PubMed2001

3,9-Bis(dicyanomethylene)-2,4,8,10-tetrathiaspiro[5.5]undecane.

The title compound, 2,2'-(2,4,8,10-tetrathiaspiro[5.5]undecane-3,9-diylidene)bis(propanedinitrile), C(13)H(8)N(4)S(4), has been designed and synthesized for use as a potential new organic molecular electronic material. The spiro-annulated structure has twofold symmetry and is formed by two equal push-pull ethylene units, with the cycloalkylthio groups as electron donors and the cyano groups as electron acceptors. The intermolecular S.N non-bonded separation within a layer in the lattice is 3.296 (6) A, indicating a strong intermolecular interaction between the cyano groups and the S atoms, while the S atoms in two neighbouring molecules have a shortest S.S contact of 3.449 (3) A. In addition, attractive C-H.N and C-H.S interactions bridge adjacent molecules either within a layer or between layers. In short, these four types of intermolecular interactions combine to form an extended three-dimensional network in the lattice, resulting in a highly ordered array of molecular packing.

Journal Article↗
PubMed2001

Determination of the enhancing action of HSP90 on neuronal nitric oxide synthase by EPR spectroscopy.

Recent studies showed that heat shock protein 90 (HSP90) enhances nitric oxide (NO) synthesis from endothelial and neuronal NO synthase (eNOS and nNOS, respectively). However, these findings were based on indirect NO measurements. Moreover, although our previous studies showed that the action of HSP90 involves increased Ca(2+)/calmodulin (Ca(2+)/CaM) binding, quantitative measurements of the effect of HSP90 on CaM binding to nNOS have been lacking. With electron paramagnetic resonance spectroscopy, we directly measured NO signals from purified nNOS. HSP90 augmented NO formation from nNOS in a dose-dependent manner. Tryptophan fluorescence-quenching measurements revealed that HSP90 markedly reduced the K(d) of CaM to nNOS (0.5 +/- 0.1 nM vs. 9.4 +/- 1.8 nM in the presence and absence of HSP90, P < 0.01). Ca(2+) ionophore triggered strong NO production from nNOS-transfected cells, and this was significantly reduced by the HSP90 inhibitor geldanamycin. Thus these studies provide direct evidence demonstrating that HSP90 enhances nNOS catalytic function in vitro and in intact cells. The effect of HSP90 is mediated by the enhancement of CaM binding to nNOS.

Benzoquinones↗
PubMed2001

Pre- and postsynaptic inhibition by nociceptin in guinea pig small intestinal myenteric plexus in vitro.

Actions of nociceptin on electrical and synaptic behavior of morphologically and neurochemically identified neurons in the guinea pig duodenal myenteric plexus were studied with conventional techniques. Nociceptin hyperpolarized the membrane potential in 104 of 121 AH-type and 28 of 51 S-type neurons with an EC(50) of 11.9 +/- 1.2 nM. Increased K(+) conductance accounted for the hyperpolarizing responses that were blocked by pertussis toxin and unaffected by naloxone. The selective opioid receptor-like (ORL)(1) receptor antagonist [Phe(1)-psi(CH(2)-NH)-Gly(2)]nociceptin(1--13)-NH(2) suppressed the nociceptin-evoked responses while behaving like a partial agonist. The nonselective ORL(1) antagonist naloxone benzoylhydrazone competitively suppressed nociceptin actions with a pA(2) value of 5.8. Nociceptin acted at presynaptic inhibitory receptors to suppress fast excitatory nicotinic postsynaptic potentials in 25 of 30 neurons (EC(50) = 22.5 +/- 4.4 nM) and slow synaptic excitation in 38 of 45 neurons (EC(50) = 15.1 +/- 1.6 nM). Presynaptic inhibitory action of nociceptin was unaffected by naloxone and was antagonized by [Phe(1)-psi(CH(2)-NH)-Gly(2)]nociceptin(1--13)-NH(2) or naloxone benzoylhydrazone. The results suggest that nociceptin acts both pre- and postsynaptically by activating an ORL(1) receptor that is distinct from typical naloxone-sensitive opioid receptors.

Animals↗
PubMed2001

Extraction kinetics of phenol with N, N-di (1-methyl-heptyl) acetamide-kerosene using hollow fiber membrane extractor.

Extraction kinetics of phenol with N,N-di(1-methyl-heptyl) acetamide-kerosene using hollow fiber membrane extractor has been studied. The rate regularities and kinetic types of forward and backward extraction were obtained respectively by determining the forward and backward extraction rate under various experimental conditions. The mass transfer mechanism was discussed. Both the forward and backward extraction of phenol might be controlled by diffusion processes, and the diffusion resistance for both forward and backward extraction mainly exists in aqueous phase. In addition, ways to optimize the extraction process of phenol were discussed as well.

Acetamides↗
PubMed2001

Arsenic contamination of groundwater and prevalence of arsenical dermatosis in the Hetao plain area, Inner Mongolia, China.

An investigation was carried out on arsenic contamination of groundwater and prevalence of arsenical dermatosis in the Hetao plain of Inner Mongolia Autonomous Region, China. Based on the screening of water samples from 96 randomly selected wells in this Region, two areas (Wuyuan and Alashan) were chosen as highly contaminated areas because arsenic in the water samples was higher than 50 microg/l. Arsenic was measured using an arsenic silver diethyl dithiocarbamate method for 326 water samples from all the wells in these areas. The results show arsenic contaminated groundwater from tubule-type wells of depths about 15-30 m was serious compared with open-type wells where depth is about 3-5 m. In the Wuyuan area, 96.2% of water samples from tubule-type wells contained arsenic above 50 microg/l and 69.3% in Alashan area; the highest value was 1354 microg/l and 1088 microg/l, respectively. In these two areas, a health survey was carried out for arsenical dermatosis. The results show the prevalence of arsenical dermatosis in the Wuyuan area was 44.8%, higher than 37.1% prevalence of arsenical dermatosis in the Alashan area. The prevalence of arsenical dermatosis was highest in the over 40-year-old age group. There was no sex difference in the prevalence. Further investigation is needed to clarify the actual situation of arsenic pollution of groundwater in Inner Mongolia, China in order to reduce the adverse health effect among residents exposed to arsenic.

Adolescent↗
PubMed2001

[The scleral buckling combined with argon laser photocoagulation for retinal detachment].

OBJECTIVE: To investigate the effects and indications of postoperative photocoagulation with argon laser instead of intraoperative cryotherapy for closing the retinal breaks during the scleral buckling operation in rhegmatogenous retinal detachment (RD) METHODS: An analysis of clinical data in 35 RD cases (38 eyes) was conducted retrospectively. These cases were treated by argon laser photocoagulation after the scleral buckling surgery from December, 1999 to April, 2000 in our hospital. RESULTS: Thirty-eight eyes were followed-up from 3 to 6 months. In 36 of 38 eyes, the retina was reattached after surgery (94.7%). Postoperatively, thirty-one eyes (86. 1%) had a visual acuity (VA) of >or= 0.05 and the best VA was 1.0. CONCLUSIONS: The method using postoperative photocoagulation instead of cryotherapy during scleral buckling surgery can simplify the surgical manipulations, alleviate the vessel damage of the retina and uvea and reduce the postoperative complications. It is a practicable way to treat RD. Surely, photocoagulation can not totally replace cryotherapy. Proper selection of indications and avoidance of complications of photocoagulation can elevate the successful rate of reattachment of RD, and promote the recovery of visual function.

Adolescent↗
PubMed2001

[Production and detection of monoclonal anti-idiotype antibodies against Vibrio anguillarum].

Vibrio anguillarum is the pathogenic bacteria of Vibriosis, which is an infectious disease found in various fish species. Seven monoclonal anti-idiotype antibodies(mAb2) were raised against mAb1 4A6. Identification of subgroup showed that 1H5, 1D1, 2B12 and 2F12 belonged to IgG2b, 2H12 and 1H12 to IgG2a and 1E10 to IgG3. The titers of these mAb2 ascites were 1 x 10(-4)-1 x 10(-6). The capacity of the mAb2 to inhibit the binding between mAb1 and antigen was investigated with the competitive inhibition ELISA. The results showed that 1D1, 1E10 1H5 and 2H12 mAb2 were able to inhibit this binding. Another experiment demonstrated that four mAb2(1D1, 1E10, 1H5 and 2H12) might induce Balb/c mice to produce Ab3 and these Ab3 competed the same antigen epitopes with Ab1. These results indicate that mAb2(1D1, 1E10, 1H5 and 2H12) are likely to represent internal image of antigen and belong to Ab2 beta. They might be employed to induce antibodies against pathogenic epitopes of V. anguillarum in vivo so as to gave the safe and effective vaccine.

Animals↗
PubMed2001

[Studies on the determination of serum thymic factor by capillary zone electrophoresis].

The determination of serum thymic factor (FTS) by capillary zone electrophoresis was described. The optimum ionic concentration and the pH value of buffer system was examined through orthogonal analysis. The optimum conditions found to be 0.05 mol/L sodium tetraborate, 0.01 mol/L phosphate buffer at pH 8.70, working voltage at 12 kV and wavelength at 200 nm. The recoveries for FTS standards added to the bovine serum albumin and serum samples were measured. The values of the relative standard deviation(RSD), recovery and detection limit were 7.62%, 80.28% and 40 ng/ml respectively. The development of this method provide another way for studying the relations of zinc and thymic factor and investigating the effect of zinc on immune function.

Electrophoresis, Capillary↗