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Biomedical subjects

W Wang

Publications and source records attributed to W Wang.

At least 865 records · Page 48Linked to original sources
PubMed1998

Neuronal nitric oxide synthase is expressed in principal cell of collecting duct.

We used the RT-PCR technique and immunocytochemical methods to determine the expression of endothelial nitric oxide synthase (eNOS) or neuronal nitric oxide synthase (nNOS) in the cortical collecting duct (CCD) in rats on high-K+ diet. The microdissected CCDs of the rat kidney were lysed, and RT-PCR was carried out using rat nNOS and eNOS gene-specific primers. Southern analysis showed the presence of mRNA of nNOS but not eNOS in the CCD. The presence of nNOS in the CCD was further confirmed by light microscopy. We used the polyclonal nNOS antibody in immunocytochemical studies of the isolated CCD. We found that immunoreactivity to nNOS was present in the CCD and heterogeneous with positive and negative immunostaining. We performed the immunocytochemical studies in the split-open CCD and found that the immunoreactivity to nNOS was detected only in principal cells but not in intercalated cells. We conclude that nNOS is expressed in the rat CCD in rats on high-K+ diet. The presence of nNOS in the CCD is heterogeneous and mainly located in principal cells.

Animals↗
PubMed1998

Carrier-mediated GABA release activates GABA receptors on hippocampal neurons.

gamma-Aminobutyric acid (GABA) transporters are electrogenic and sodium-dependent and can operate in reverse when cells are depolarized or when there is reversal of the inward sodium gradient. However, the functional relevance of this phenomenon is unclear. We have examined whether depolarization induced by a physiologically relevant increase in extracellular [K+] leads to sufficient amounts of carrier-mediated GABA release to activate GABAA receptors on neurons. Patch-clamp recordings were made from rat hippocampal neurons in culture with solutions designed to isolate chloride currents in the recorded neuron. Pressure microejection was used to increase extracellular [K+] from 3 to 12 mM. After blockade of vesicular GABA release by removal of extracellular calcium, this stimulus induced a large conductance increase in hippocampal neurons [18.9 +/- 6.8 (SD) nS; n = 16]. This was blocked by the GABAA receptor antagonists picrotoxin and bicuculline and had a reversal potential that followed the Nernst potential for chloride, indicating that it was mediated by GABAA receptor activation. Similar responses occurred after block of vesicular neurotransmitter release by tetanus toxin. GABAA receptors also were activated when an increase in extracellular [K+] (from 3 to 13 mM) was combined with a reduction in extracellular [Na+] or when cells were exposed to a decrease in extracellular [Na+] alone. These results indicate that depolarization and/or reversal of the Na+ gradient activated GABA receptors via release of GABA from neighboring cells. We found that the GABA transporter antagonists 1-(4, 4-diphenyl-3-butenyl)-3-piperidinecarboxylic acid hydrochloride (SKF89976A; 20-100 microM) and 1-(2-([(diphenylmethylene)amino]oxy)ethyl) -1, 2, 5, 6 - tetrahydro - 3 - pyridine - carboxylic acid hydrochloride (NO-711; 10 microM) both decreased the responses, indicating that the release of GABA resulted from reversal of the GABA transporter. We propose that carrier-mediated GABA release occurs in vivo during high-frequency neuronal firing and seizures, and dynamically modulates inhibitory tone.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1998

Modified ultrafiltration in paediatric cardiopulmonary bypass.

Cardiopulmonary bypass (CPB), a nonphysiological procedure, is associated with haemodilution and the inflammatory response, causing the accumulation of body water and organ dysfunction. The purpose of this study was to evaluate the efficacy of modified ultrafiltration. Forty paediatric patients undergoing cardiac operations were randomized into a control group and a modified ultrafiltration group. Blood cells, protein and cytokine concentrations were recorded for 24 h postoperatively. As the fluid was removed at 50 ml/min, both blood cells and protein were concentrated by modified ultrafiltration (p < 0.001). The tumour necrosis factor (TNF)-alpha concentration was increased and interleukin-8 (IL-8) and endothelin (ET) concentrations were unaltered after ultrafiltration. After correction for albumin, TNF-alpha concentration changed little, and IL-8 and ET concentrations (36.75 +/- 12.35, 42.89 +/- 15.54) were decreased significantly (21.47 +/- 13.87, 26.06 +/- 12.54) after ultrafiltration. Modified ultrafiltration is an effective method for removing excess tissue fluid and concentrating blood after CPB. This technique can also filter out some cytokines.

Blood Cell Count↗
PubMed1998

Inner ear and maternal reproductive defects in mice lacking the Hmx3 homeobox gene.

The Hmx homeobox gene family is of ancient origin, being present in species as diverse as Drosophila, sea urchin and mammals. The three members of the murine Hmx family, designated Hmx1, Hmx2 and Hmx3, are expressed in tissues that suggest a common functional role in sensory organ development and pregnancy. Hmx3 is one of the earliest markers for vestibular inner ear development during embryogenesis, and is also upregulated in the myometrium of the uterus during pregnancy. Targeted disruption of the Hmx3 gene results in mice with abnormal circling behavior and severe vestibular defects owing to a depletion of sensory cells in the saccule and utricle, and a complete loss of the horizontal semicircular canal crista, as well as a fusion of the utricle and saccule endolymphatic spaces into a common utriculosaccular cavity. Both the sensory and secretory epithelium of the cochlear duct appear normal in the Hmx3 null animals. The majority of Hmx3 null females have a reproductive defect. Hmx3 null females can be fertilized and their embryos undergo normal preimplantation development, but the embryos fail to implant successfully in the Hmx3 null uterus and subsequently die. Transfer of preimplantation embryos from mutant Hmx3 uterine horns to wild-type pseudopregnant females results in successful pregnancy, indicating a failure of the Hmx3 null uterus to support normal post-implantation pregnancy. Molecular analysis revealed the perturbation of Hmx, Wnt and LIF gene expression in the Hmx3 null uterus. Interestingly, expression of both Hmx1 and Hmx2 is downregulated in the Hmx3 null uterus, suggesting a hierarchical relationship among the three Hmx genes during pregnancy.

Alleles↗
PubMed1998

N-acetylcysteine as an antidote in methylmercury poisoning.

Methylmercury is a ubiquitous environmental pollutant and potent neurotoxin. Treatment of methylmercury poisoning relies almost exclusively on the use of chelating agents to accelerate excretion of the metal. The present study demonstrates that oral administration of N-acetylcysteine (NAC), a widely available and largely nontoxic amino acid derivative, produces a profound acceleration of urinary methylmercury excretion in mice. Mice that received NAC in the drinking water (10 mg/ml) starting at 48 hr after methylmercury administration excreted from 47 to 54% of the 203Hg in urine over the subsequent 48 hr, as compared to 4-10% excretion in control animals. When NAC-containing water was given from the time of methylmercury administration, it was even more effective at enhancing urinary methylmercury excretion and at lowering tissue mercury levels. In contrast, excretion of inorganic mercury was not affected by oral NAC administration. The ability of NAC to enhance methylmercury excretion when given orally, its relatively low toxicity, and is wide availability in the clinical setting indicate that it may be an ideal therapeutic agent for use in methylmercury poisoning.

Acetylcysteine↗
PubMed1998

Adult carboxypeptidase E-deficient fat/fat mice have a near-total depletion of brain CCK 8 accompanied by a massive accumulation of glycine and arginine extended CCK: identification of CCK 8 Gly as the immediate precursor of CCK 8 in rodent brain.

Cholecystokinin (CCK) amide concentrations were reduced over 85% in all the major brain regions of carboxypeptidase E (Cpe)(fat)/Cpe(fat) mice in comparison to control mice. Using an radioimmunoassay (RIA) specific for glycine-extended CCK (CCK Gly), low levels of CCK Gly were detected in control (0.65 ng/g tissue) and were even lower in Cpe(fat)/Cpe(fat) (0.246 ng/g) mice brain extracts. After treatment with carboxypeptidase B, the level of CCK Gly in Cpe(fat)/Cpe(fat) in these brain extracts was elevated to 33.5 ng/g, about 51-fold higher than in control. On gel-filtration chromatography and high-performance liquid chromatography (HPLC), this material coeluted with CCK 8 Gly. These results demonstrate that CPE is required for the correct processing of arginine- and glycine-extended CCK in all major regions of the mouse brain. These results support the hypothesis that CCK 8 Gly is the immediate precursor of CCK 8 amide in mouse brain, not larger amidated forms like CCK 22 or CCK 33.

Animals↗
PubMed1998

G protein signalling pathways in myometrium: affecting the balance between contraction and relaxation.

Heterotrimeric G proteins are actively involved in intracellular signalling in the myometrium and play important roles in regulating myometrial contraction and relaxation. Increases in intracellular calcium can be induced by agents that stimulate uterine contractions. In a number of instances, these increases in intracellular calcium are attributed to stimulation of phospholipase C by either G alpha or G betagamma subunits as a result of activation of G protein-coupled plasma membrane receptors. This mechanism also stimulates calcium entry through calcium release-activated channels, either directly or indirectly. Thus, while phospholipase C can be activated by other pathways and calcium can enter myometrial cells through other channels, G proteins play a major role in these processes. Similarly, activation of protein kinase A and protein kinase C are consequences of G protein activation. Protein kinase A and protein kinase C exert a number of regulatory influences on phospholipase C, ion channel activity and other processes in the myometrium. The mitogen-activated protein kinase pathway can also be activated directly or indirectly by the action of G proteins in myometrium. Responsiveness to G proteins can be altered during pregnancy and depends on the relative expression of all of the components of the signalling pathways involved. The balance between G protein-mediated stimulatory and inhibitory signalling pathways has important consequences for the control of myometrial contractile activity.

Adenylyl Cyclases↗
PubMed1998

Far UV photolysis of uracil and cytosine in phosphate solution.

The photolysis of nucleobases, nucleosides and nucleotides (NA) was enhanced by phosphate under irradiation of medium pressure mercury lamp (MPML). Uracil and Cytosine in phosphate solution were selected to study the mechanism of phosphate effect. Photoproducts were produced in the irradiated uracil and cytosine of phosphate solution which have been isolated by anion exchange resin. Ultraviolet irradiation (190-220nm) of uracil in 0.05 mol/dm3 phosphate buffered solution at pH 8-9 leads to the production of a novel compound C4H5N2O6P which has been identified by UV, 1H-NMR spectroscopy and LC/MS/MS. The formation mechanism of the photoproduct and the kinetics were studied.

Amino Acids↗
PubMed1998

Chiral beta-particles interact differentially with enantiomers--theory to solve a long-existing debate.

The crucial organic molecules associated with life are chiral. Beta irradiation on the causal origin for the great preference of biomolecules in life over their corresponding enantiomers has been extensively studied. The left helical beta electrons, with spin and momentum antiparallel, should propagate with different velocities in the two enantiomers. Much effort has been done to investigate possible mechanism of inducing asymmetry, using longitudinally polarized beta rays to decompose asymmetrically racemic mixtures of biomolecules, but the results were inconclusive and contradictory. In this paper we present our viewpoint that the direct inelastic scattering between polarized electrons and chiral molecules is the dominant one in producing the asymmetry of cross sections for both beta+ and beta- irradiation. The inelastic cross section sigma (0 --> n) is dependent on the dipole strength Dn, rotatory strength Rn of the molecule. According to the theoretical study, the asymmetry of cross section F is approximately 10(-6) and it depends on the sign of Rn+(L type). When Rn+ > 0, sigma D > sigma L, it favors L-amino acid surviving. When Rn+ < 0, sigma D < sigma L, it favours D-amino acid surviving. Our results show that helical electrons do distinguish between molecules of opposite chirality, and the disputation and suspicion about the controversial experimental results between Garay and Darge & Thiemann can be explained.

Amino Acids↗
PubMed1998

HPLC analysis of isoflavonoids and other phenolic agents from foods and from human fluids.

A fast, precise and selective diode array HPLC method is presented for the extraction and analysis of soy isoflavonoids from foods and from human urine, plasma, and breast milk in support of mechanistic and epidemiologic studies assessing the potential cancer protective role of soya or isoflavones. Solid phase or solvent extraction was chosen for isolation, and enzymatic or acid hydrolysis procedures were used for aglycone production depending on the matrix to be analyzed. C-18 reversed-phase HPLC was applied to selectively separate and quantitate daidzein (1), glycitein (3), and genistein (4), including their malonyl (a) and acetyl (b) esters, and their mammalian metabolites equol (6) and O-desmethylangolensin (7), as well as formononetin (2), biochanin-A (5), and coumestrol (8) using a gradient elution system. UV absorbance scans and authentic standards were applied for identification purposes, additional to fluorometric monitoring, electrochemical detection, and GC/ MS analysis after trimethyl silylation. Detection limits of 20-microl injections were found to be 1.09, 0.53, 3.28, and 1.00 pmoles for daidzein, genistein, equol, and O-desmethylangolensin (DMA), respectively, by monitoring at the individual compound's absorption maximum. The proposed method was applied to monitor isoflavone levels in soy foods and in human plasma, urine and breast milk after challenge with roasted soybeans. Implications of the presented results on the potential activity of isoflavones to prevent cancer by exposing newborn infants to these agents are discussed.

Chromatography, High Pressure Liquid↗
PubMed1998

Radioimmunoassay determination of formononetin in murine plasma and mammary glandular tissue.

Formononetin is an isoflavonoid phytoestrogen found in animal fodder and in certain human foodstuffs. Since high consumption of formononetin has been implicated in infertility among ruminants and may relate to human breast cancer, we developed a radioimmunoassay method and applied it to quantitate formononetin in murine plasma and mammary glandular tissue for animal model studies. The radioimmunoassay utilized an antiserum raised in rabbits following immunization with formononetin-7-O(carboxymethyl)ether coupled to bovine serum albumin. The tracer was an [3H]leucine derivative of formononetin synthesized by mixed anhydride reaction between formononetin-7-O(carboxymethyl)ether and [3H]leucine. The bound and free forms of formononetin were separated by adding dextran-coated charcoal. This radioimmunoassay procedure enabled the quantification of 4 ng/ml of plasma or 50 pg/mg of mammary tissue, and the antiserum showed no marked cross-reaction with the reactants tested. The reliability and reproducibility of the assay were demonstrated by intra- and inter-assay variation that was 6.5% and 11.9%, respectively. This radioimmunoassay was compared with a high-performance liquid chromatographic method by determining concentrations of formononetin in ethanol extracts of red clover. Good correlation existed between the radioimmunoassay and high-performance liquid chromatographic method (r = 0.980), but the radioimmunoassay values were on the average 5% higher than high-performance liquid chromatographic values. Furthermore, we used the radioimmunoassay to assess the pharmacokinetics of formononetin in castrated female BALB/c mice after a single subcutaneous injection at 40 mg/kg dose. The sensitivity of the radioimmunoassay permitted the detection of a prolonged elimination phase for formononetin in both plasma and mammary glandular tissues, with mean elimination half-lives of 2 and 2.5 hr, respectively. A specific, sensitive, and fast radioimmunoassay procedure has been developed for the determination of formononetin in murine plasma and mammary glandular tissue. We conclude that the presented radioimmunoassay is a valid alternative method to the quantification of formononetin in biological fluids and/or plants.

Animals↗
PubMed1998

[The role of fetus decalcified bone matrix (FDBM) in inducing pure titanium-bone implant integration].

Because of its high biological compatibility, titanium has been a good biomaterial. The implanted artificial bone made from titanium can contact with the vital and mature osseous tissue directly within 3-6 months, the so-called osteointergration. In order to promote the process of osteointergration, FDBM of rabbit was prepared and was combined with pure titanium so as to speed up osteointergration. The study focused on bone density, bone intergration rate, new bone growth rate around the pure titanium, and the Ca2+ and PO(4)3- density of titanium-bone interface. A control group of pure titanium inplant without FDBM was set up. The results showed FDBM had no antigenicity. It could induce and speed up the new bone formation at titanium-bone interface. The titanium-bone intergration time was within 2 months. It was suggested that there were more bone morphogenesis protein (BMP) or other bone induction and bone formation factors in brephobone than that in child and adult bone. As a kind of bone induction material, FDBM was easy prepared, cheap in price, easy to storage, no antigenicity and obvious bone-inductive function.

Animals↗
PubMed1998

[Determination of intracellular calcium ions in fibroblasts of contractured scar].

Free calcium ions, as a kind of message-transport substance, is important in cellular activity such as cell movement, cell differentiation and cell proliferation. In order to investigate the relationship between free calcium ions and scar contracture, the fibroblasts which originated from hypertrophic scar, keloid and normal skin were used as the experimental target. The fibroblasts from 4th-6th generations of different sources were used; Then the intracellular free calcium ions concentrations were measured respectively by the fluorescent Ca2+ indicator Fura-2/AM and Image analysis system. The results showed that the level of Ca2+ in fibroblasts of hypertrophic scar was higher than that in keloid and normal skin (P < 0.01). There was no significant difference between the level of Ca2+ in keloid and in normal skin. The conclusion was that the concentration of intracellular free calcium ions played an important role in the scar contract, but the exact mechanism was still unclear and required further study.

Biological Transport, Active↗
PubMed1998

Determination of S/R ratio of mephenytoin in human urine by chiral HPLC and ultraviolet detection and its comparison with gas chromatography.

AIM: To improve HPLC method for rapid determination of urinary S/R-ratio of mephenytoin, a widely used metabolic index for cytochrome P-450 2C19 (CYP2C19) activity. METHODS: Aliquots of 0-8-h urine sample after dosing racemic mephenytoin 100 mg underwent one-step extraction with dichloromethane. Analysis was performed on a chiral column (250 mm x 4 mm, 5 microns) at lambda = 207 nm. The eluent was a mixture of acetronitrile and water containing both 0.1% glacial acetic acid and 0.2% triethylamine (14:86, vol/vol) at a flow-rate of 0.9 mL.min-1. RESULTS: The enatiomers of mephenytoin in urine were well separated within 9 min. A linear correlation was observed between 50-5000 micrograms.L-1 with the detection limit of 12.5 micrograms.L-1 for both enantiomers of mephenytoin. This HPLC analysis was comparable to gas chromatography in accuracy and sensitivity, but with much shorter retention time and better resolution. CONCLUSION: The present HPLC method is good for rapid determination of the ability of subjects to hydroxylate S-mephenytoin after oral administration of the racemic drug.

Aryl Hydrocarbon Hydroxylases↗
PubMed1998

[Expression of proliferating cell nuclear antigen in hemangioma and vascular malformations].

OBJECTIVE: This study was to compare the proliferative activity of endothelial cells of various types of hemangiomas and vascular malformations in different phases. METHODS: Forty-one specimens of hemangiomas or vascular malformations from biopsy or operative resection were stained with immunohistochemical method for proliferating cell nuclear antigen (PCNA). The proliferative activity of endothelial cells was expressed by labeling index (LI). RESULTS: PCNA was positive in all of proliferating strawberry, composite hemangiomas and some of cavernous hemangiomas in children. PCNA was negative in all of adult involuted hemangiomas and some of cavernous hemangiomas in children. There were no statistically significant differences in labeling indices of PCNA between various types of proliferating hemangiomas in children while there were statistically significant differences between child proliferating hemangiomas and involuted hemangiomas in children or all of adult hemangiomas. CONCLUSION: The results may be helpful to answer some bewildering questions about hemangiomas and provide a basis for new classification.

Adult↗