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Biomedical subjects

G Wang

Publications and source records attributed to G Wang.

At least 505 records · Page 28Linked to original sources

[Simulated experiment of hemodynamics changes in aorta during high energetic trauma through abdomen].

In order to study the mechanism of remote effect of missile wound, we designed and carried out a simulated experiment. The change of hemodynamics in aorta was examined with a pressure transducer during gunshot and then calculated. The results revealed superhigh pressure, high speed of pressure rising, accelerated velocity of blood flow, substantial pressure difference and profuse blood flow in aorta during gunshot.

Abdominal Injuries↗

[A comparison of amino acid composition of proteins from thermophiles and mesophiles].

The thermophilic feature of thermophiles are due to the high thermostability of their proteins. In order to investigate the mechanism of the high thermostability of thermophilic proteins and compare the difference of amino acid composition between thermophilic and mesophilic proteins, 110 pairs of homologous protein sequences from thermophiles and mesophiles respectively are collected and the amino acid composition, percentage of hydrophobic amino acids, hydropathy index, percentage of charged amino acids of the two protein groups are compared. It reveals that the two groups have evidently different composition in several kinds of amino acids, and thermophilic proteins have higher average hydropathy and charged amino acids composition. A comparison of the aliphatic indices between the two protein groups indicates that the reason that thermophilic proteins have higher aliphatic index is that thermophilic proteins have higher leucine composition, and thus sheds doubt on the validity of aliphatic index. It shows that it is possible to disclose some general rules with regard to the mechanism of protein thermostability by comparing a large amount of protein sequence data.

Amino Acids↗

[Analysis of chemical components from Pheretima].

The total fat, total nitrogens, free amino acids, inorganic elements of Pheretima have been determined. It will provide scientific evidence for the establishment of quality standards of pheretima.

Amino Acids↗

[Experimental study on the effect of abstinence with herbal preparation qingjunyin].

The effect of herbal preparation Qingjunyin (QJY) on the animal model of drug dependence was studied. After the morphinistic models of white rats and mice were made by ever increasing doses of morphine each time, the animals were divided into large dosage QJY group, small dosage QJY group, sustained morphine group and control group. The withdrawal symptoms were observed after naloxone was given to the animals intraperitoneally. The results showed that QJY groups withdrawal symptoms of the addicted white rats were alleviated, the number of jumps of the addicted mice was reduced, the scores of the principal symptoms of the animals were decreased, while the body weights of the animals were gained, comparing with the control group, the difference was significant (P < 0.01). The effects of QJY enhanced in the large dosage group, which suggests QJY has abstinence effects.

Animals↗

[Determination of trace selenium and tellurium in steel by hydride-generation nondispersive atomic fluorescence spectrometry].

A method was developed for the determination of trace Se and Te in steel by ion-exchange-separation hydride-generation nondispersive atomic fluorescence spectrometry. 40% (V/V) chlorhydric acid was selected as the best medium for hydride reaction. The detection limits of Se and Te are 0.27 and 0.33 ng/mL, relative standard deviations are 6.99 and 6.11%, respectively. The recovery was in the range of 96.7-102.0%. The results are satisfactory.

English Abstract↗

[Analysis of the properties of plasma produced by pulsed laser ablation of metal Al at different ambient pressures].

A time and space resolved diagnostic technique was used to study the properties of plasma produced by a pulsed laser on an Al target irradiated at different ambient pressures. The velocities of projected particles were measured. The projected ranges for 1-10keV AI2+ ions ejecting into ambient gas at different pressures were calculated in theory. The calculated results can explain the observed experimental phenomena successfully.

English Abstract↗

Confocal microscopy assessment of lymphoid tissues with follicular hyperplasia from patients infected with human immunodeficiency virus type 1.

OBJECTIVE: To characterize human immunodeficiency virus (HIV) infection of lymphoid tissues during follicular hyperplasia. METHODS: We examined 10 tonsil/adenoid, 3 parotid lymphoepithelial cyst, and 7 lymph node specimens that had been surgically removed from 13 patients infected with HIV-1. Characteristics of productive HIV-1 infection were assessed using immunocytochemistry for HIV-1 p24. Cellular colocalization was determined with the aid of a confocal microscope using double immunofluorescent staining for HIV-1 p24 and cell-specific markers. RESULTS: All specimens showed follicular hyperplasia. Using confocal microscopy with three-dimensional reconstruction, HIV-1 p24 was seen to be "intimately" colocalized with CD21 within the germinal centers. While lymphoid follicles were generally hyperplastic, only a subset of these follicles contained HIV-1 p24. Occasional HIV-1-expressing mononuclear cells identified outside follicles stained for CD68 or CD3. CONCLUSIONS: The differential involvement of hyperplastic follicles by HIV-1 within individual lymphoid tissues and the intimate colocalization of HIV-1 p24 and CD21 suggest that infected follicular dendritic cells may be an important reservoir of HIV-1 during follicular hyperplasia.

Adult↗

Homology modeling of tissue-type plasminogen activator K1 domain and studies on the interactions between kringles and lysine.

The 3-D structure of t-PA K1 domain was predicted by the method of homology modeling. The putative lysine-binding pockets of t-PA K1, UK K, PLG K1 and K4 were determined by superimposing their 3-D structures to that of t-PA K2 domain, of which the lysine-binding pockets had been revealed previously. After that the key residues of lysine-binding pockets of kringles were identified. The structural analyses showed that both the electrostatic potential and hydrophobic complementarity were well matched between lysine and the binding pockets of t-PA K2, PLG K1 and K4, but for t-PA K1 and UK K domains the complementarity did not match well in one or both respects. It is proposed that this is the reason that t-PA K1 and UK K domains do not bear the ability to bind lysine. With the respect of improving the affinity for fibrin, new types of mutants of t-PA K1 and UK K domains were designed, and structural changes caused by mutation were predicted by simulating the residue replacements. The mutant structural models demonstrated that the molecular design was reasonable.

Amino Acid Sequence↗

Molecular genetic probing of energy coupling by the yeast plasma membrane proton pump.

Genetic probing of PMA1, which encodes the plasma membrane H(+)-ATPase, has highlighted the putative role of the N-terminal half of the enzyme in the coupling process. Recent second-site suppressor studies indicate that significant interactions occur between the region near the site of phosphorylation, stalk segment 3 (S3), and the N-terminal transmembrane segments. Saturation mutagenesis was used to explore I183 in S2, which partially uncouples proton transport when converted to alanine. Numerous substitutions could be made at this position. However, stable substitutions with Arg, Tyr or Asn were often accompanied by second-site mutations at the extreme C-terminus, suggesting a close interaction between these regions. Several mutations in the putative stalk domain are known to alter coupling, and scanning glycine and proline mutagenesis was used to probe the predicted alpha-helical character of the stalk segments. The results indicate that the introduction of proline or glycine in S2, S4 or S5, was highly disruptive to enzyme function often resulting in cell death. Similar substitutions in stalk 3 yielded viable but significantly altered enzymes. These results suggest that the helical properties of these segments may be important for catalysis. Finally, the stalk region has been modeled as a helical bundle, which helps account for the effects of specific perturbations in this region.

Adenosine Triphosphate↗

In vivo measures of cochlear length and insertion depth of nucleus cochlear implant electrode arrays.

Three-dimensional cochlear canal lengths, electrode array intracochlear insertion depths, and characteristic frequency ranges were estimated for 20 Nucleus implant recipients on the basis of in vivo computed tomography (CT) scans. Ultra-high resolution images were reconstructed from spiral CT data with 0.1-mm slicing and expanded attenuation scales. Canal length estimates (mean 33.01 mm; SD 2.31) were consistent with previous findings for normal human temporal bones. Intracochlear array insertion depths estimated by 3-dimensional (3-D) spiral calculations (mean 20.19 mm; SD 2.86) and by a computerized array tracking algorithm (mean 20.36 mm; SD 2.66) were not significantly different. Estimates from surgical observations were significantly longer (mean 21.03 mm; SD 2.31) because array compressions were not detectable. Characteristic frequencies at apical electrodes estimated from Greenwood's equations ranged from 387 Hz to 2,596 Hz. The results show that significant variations in cochlear anatomy and array distribution among implant patients that may impact implant performance can be reliably detected and quantified by using in vivo high-resolution CT and 3-D reconstructions.

Adult↗

Highly sensitive time-resolved fluoroimmunoassay of human immunoglobulin E by using a new europium fluorescent chelate as a label.

A new europium fluorescent chelate, 4,4'-bis(1",1",1",2",2",3",3"-heptafluoro-4",6"-hexanedione-6"-yl) - chlorosulfo-o-terphenyl (BHHCT)-Eu3+, was used as a label for highly sensitive time-resolved fluoroimmunoassay of human IgE. Two assay formats were employed in the analysis. In the first format, an immunoconjugate of rabbit anti-human IgE antibody-human IgE-biotinylated goat anti-human IgE antibody-BHHCT-Eu(3+)-labeled SA (or BHHCT-Eu(3+)-labeled BSA-SA; BSA, bovine serum albumin; SA, streptavidin) was used for measurement. The method gives the detection limits of 3.6 x 10(-2) IU/ml (labeled SA) and 1.1 x 10(-2) IU/ml (labeled SA-BSA). In the second format, an immunoconjugate of goat anti-human IgE antibody-human IgE-rabbit anti-human IgE antibody-biotinylated goat anti-rabbit IgG antibody-BHHCT-Eu(3+)-labeled SA (or BHHCT-Eu(3+)-labeled BSA-SA) was used for measurement. The detection limits of these methods are 3.0 x 10(-3) IU/ml (labeled SA) and 1.5 x 10(-3) IU/ml (labeled BSA-SA). The above detection limits are one to two orders of magnitude lower than those of the conventional radioimmunoassay and enzyme immunoassay. The CV of the present method is less than 7%, and the recovery is in the range of 85-105% for serum samples.

Animals↗

Conformational changes of P-glycoprotein by nucleotide binding.

P-glycoprotein (Pgp) is a membrane protein that transports chemotherapeutic drugs, causing multidrug resistance in human cancer cells. Pgp is a member of the ATP-binding cassette superfamily and functions as a transport ATPase. It has been suggested that the conformation of Pgp changes in the catalytic cycle. In this study, we tested this hypothesis by using limited proteolysis as a tool to detect different conformational states trapped by binding of nucleotide ligands and inhibitors. Pgp has high basal ATPase activity; that is, ATP hydrolysis by Pgp is not rigidly associated with drug transport. This activity provides a convenient method for studying the conformational change of Pgp induced by nucleotide ligands, in the absence of drug substrates which may generate complications due to their own binding. Inside-out membrane vesicles containing human Pgp were isolated from multidrug-resistant SKOV/VLB cells and treated with trypsin in the absence or presence of MgATP, Mg-adenosine 5'-[beta,gamma-imido]triphosphate (Mg-p[NH]ppA) and MgADP. Changes in the proteolysis profile of Pgp owing to binding of nucleotides were used to indicate the conformational changes in Pgp. We found that generation of tryptic fragments, including the loop linking transmembrane (TM) regions TM8 and TM9 of Pgp, were stimulated by the binding of Mg-p[NH]ppA, MgATP and MgADP, indicating that the Pgp conformation was changed by the binding of these nucleotides. The effects of nucleotides on Pgp conformation are directly associated with the binding and/or hydrolysis of these ligands. Four conformational states of Pgp were stabilized under different conditions with various ligands and inhibitors. We propose that cycling through these four states couples the Pgp-mediated MgATP hydrolysis to drug transport.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Infrared spectroscopy of human apolipoprotein fragments in SDS/D2O: relative lipid-binding affinities and a novel amide I assignment.

Infrared absorption spectra are reported for six apolipoprotein fragments in SDS/D2O. Five of the peptides correspond to proposed lipid-binding domains of human apolipoproteins [apoC-I(7-24), apoC-I(35-53), apoA-II(18-30)+, apoA-I(166-185), apoE(267-289)], and the sixth is the de novo lipid associating peptide LAP-20. The amide I infrared absorption patterns are generally consistent with predominantly helical structures (as determined previously by NMR spectroscopy and distance geometry calculations) and further suggest that apoA-I(166-185) and apoE(267-289) are bound to SDS relatively weakly in comparison to the other four peptides. The latter conclusion is also supported by the temperature dependence of the infrared spectra, as increasing temperature promotes a distinct increase in random coil structure only for apoA-I(166-185) and apoE(267-289). In addition to features readily ascribed to helices, the infrared spectra of all the peptides show absorptions in the spectral region 1630-1635 cm-1 that is usually associated with beta-structure, a motif that is clearly absent from the NMR-derived structures. Parallel difficulties also arose in the analyses of the circular dichroism spectra. We suggest that both the low-frequency infrared absorptions and the ambiguities in interpreting the CD spectra may be due to unusual structures at the peptide C-termini, involving C=O groups that form hydrogen bonds simultaneously either with two solvent molecules or with donors from the backbone (NH) and the solvent (OH). Analogous absorptions may be a general feature of solvent-exposed helices, which suggests a need for caution in assigning amide I bands below 1640 cm-1.

Amino Acid Sequence↗

The helix-hinge-helix structural motif in human apolipoprotein A-I determined by NMR spectroscopy.

The conformation of a synthetic peptide of 46 residues from apoA-I was investigated by fluorescence, CD, and 2D NMR spectroscopies in lipid-mimetic environments. ApoA-I(142-187) is mainly unstructured in water but helical in SDS or dodecylphosphocholine (DPC), although the peptide only associates with DPC at approximately the critical micellar concentration. Solution structures of apoA-I(142-187) were determined by distance geometry calculations based on 450 (in DPC-d38) or 397 (in SDS-d25) NOE-derived distance restraints, respectively. Backbone RMSDs for superimposing the two helical regions 146-162 and 168-182 are 0.98 +/- 0.22 (2.38 +/- 0.20) and 1.99 +/- 0.42 (2.02 +/- 0.21) A in DPC (SDS), respectively. No interhelical NOE was found, suggesting that helix-helix interactions between the two helical domains in apoA-I(142-187) are unlikely. Similar average, curved helix-hinge-helix structures were found in both SDS and DPC micelles with the hydrophobic residues occupying the concave face, indicating that hydrophobic interactions dominate. Intermolecular NOESY experiments, performed in the presence of 50% protonated SDS, confirm that the two amphipathic helices and Y166 in the hinge all interact with the micelle. The involvement of Y166 in lipid binding is supported by fluorescence spectroscopy as well. On the basis of all the data above, we propose a model for the peptide-lipid complexes wherein the curved amphipathic helix-hinge-helix structural motif straddles the micelle. The peptide-aided signal assignment achieved for apoA-I(122-187) (66mer) and apoA-I suggests that such a structural motif is retained in the longer peptide and most likely in the intact protein.

Amino Acid Sequence↗

HIV type 1 subtypes, coreceptor usage, and CCR5 polymorphism.

Identification of the chemokine receptors CCR5 and CXCR4 as the major coreceptors for HIV-1 entry has greatly assisted our understanding of HIV-1 pathogenesis, transmission, and tropism. However, most of our current knowledge on coreceptor usage comes from studies using HIV-1 strains or env genes derived from the genetic subtype B predominant in North America and western Europe. In this report, the coreceptor usage of 20 primary viral isolates representative of genetic subtypes A, B, C, D, E, and group O was examined. Thirty-nine full-length CCR5 sequences from individuals of diverse geographic origins were also obtained to examine the possible effect of CCR5 polymorphism on HIV-1 subtype distribution. Our results indicate that (1) CCR5 and CXCR4 serve as the two major coreceptors for viruses belonging to HIV-1 subtypes A, B, C, D, E, and group O, whereas other chemokine receptors such as CCR2b and CCR3 play only a minor role in facilitating viral entry into stimulated PBMCs; (2) the coreceptor usage is determined by the viral phenotype rather than its genotype because all NSI strains, irrespective of their subtype classification, utilize CCR5, whereas all SI strains are able to use CXCR4; and (3) there is no geographic clustering of CCR5 polymorphism in different ethnic populations, suggesting that CCR5 diversity is not the underlying explanation for differences in the spread of different HIV-1 subtypes. Therefore, the uneven worldwide distribution of HIV-1 subtypes is more likely the result of stochastic dissemination.

Amino Acid Sequence↗

Comparison of eight commercial kits for quantitation of antineutrophil cytoplasmic antibodies (ANCA).

Antineutrophil cytoplasmic antibodies (ANCA) are used as diagnostic markers for systemic vasculitis. However, the specificity and sensitivity of ANCA detection differs from centre to centre due in large part to variations in methodology. We compared 8 commercial ELISA kits and an in-house method (HM) for their specificity and sensitivity in detecting ANCA against proteinase 3 (PR3-ANCA, 7 kits) and meyloperoxidse (MPO-ANCA, 8 kits). Sera from 5 patients with systemic lupus erythematosus (SLE), 28 with Wegener's granulomatosis (WG), 22 with microscopic polyangiitis (MPA), 5 with idiopathic rapidly progressive glomerulonephritis (RPGN), and 5 healthy controls were examined by both the indirect immunofluorescence technique (IFT) and the ELISA kits. Sera from healthy controls and patients with SLE or cANCA-negative WG were shown to be PR3-ANCA negative by all 7 PR3-ANCA kits. In 25 cANCA-positive sera from WG patients, PR3-ANCA positivity ranged from 44% to 84%. An absolute concordance among the 7 kits was noted in 56% of the cANCA-positive samples. The PR3-ANCA levels in 5 of the 7 kits correlated with the cANCA titers in IFT. Sera from the healthy controls and 4 out of the 5 SLE and pANCA-negative patients were found to be MPO-ANCA negative in all 8 MPO-ANCA kits. In 20 pANCA-positive sera, MPO-ANCA positivity ranged from 25% to 75%. Thirty-five percent of MPO-ANCA-positive sera were confirmed by capture ELISA, immunoblot and inhibition assay. The concordance rate was only 30% among pANCA-positive sera in the 8 MPO-ANCA kits. No significant correlation was observed between pANCA titers and MPO-ANCA levels. The HM showed that 65% of cANCA-positive sera were PR3-ANCA positive, and 45% of pANCA-positive sera were MPO-ANCA positive. Our results indicate that the sensitivities and specificities for ANCA detection differ significantly among the commercial kits tested and underline the necessity of establishing uniform international standards for ANCA ELISA procedures in order to permit more reliable interpretation and comparison of data.

Antibodies, Antineutrophil Cytoplasmic↗

Dendritic cells retrovirally transduced with a model antigen gene are therapeutically effective against established pulmonary metastases.

Dendritic cells (DCs) are bone marrow-derived leukocytes that function as potent antigen presenting cells capable of initiating T cell-dependent responses from quiescent lymphocytes. DC pulsed with tumor-associated antigen (TAA) peptide or protein have recently been demonstrated to elicit antigen-specific protective antitumor immunity in a number of murine models. Transduction of DCs with TAA genes may allow stable, prolonged antigen expression as well as the potential for presentation of multiple, or unidentified, epitopes in association with major histocompatibility complex class I and/or class II molecules. To evaluate the potential efficacy of retrovirally transduced DCs, bone marrow cells harvested from BALB/c mice were transduced with either a model antigen gene encoding beta-galactosidase (beta-gal) or a control gene encoding rat HER-2/neu (Neu) by coculture with irradiated ecotropic retroviral producer lines. Bone marrow cells were differentiated into DC in vitro using granulocyte/macrophage colony-stimulating factor and interleukin-4. After 7 d in culture, cells were 45-78% double positive for DC phenotypic cell surface markers by FACS(R) analysis, and DC transduced with beta-gal were 41-72% positive for beta-gal expression by X-gal staining. In addition, coculture of beta-gal transduced DC with a beta-gal-specific T cell line (CTLx) resulted in the production of large amounts of interferon-gamma, demonstrating that transduced DCs could process and present endogenously expressed beta-gal. DC transduced with beta-gal and control rat HER-2/neu were then used to treat 3-d lung metastases in mice bearing an experimental murine tumor CT26.CL25, expressing the model antigen, beta-gal. Treatment with beta-gal-transduced DC significantly reduced the number of pulmonary metastatic nodules compared with treatment with Hank's balanced salt solution or DCs transduced with rat HER-2/neu. In addition, immunization with beta-gal-transduced DCs resulted in the generation of antigen-specific cytotoxic T lymphocytes (CTLs), which were significantly more reactive against relevant tumor targets than CTLs generated from mice immunized with DCs pulsed with the Ld-restricted beta-gal peptide. The results observed in this rapidly lethal tumor model suggest that DCs transduced with TAA may be a useful treatment modality in tumor immunotherapy.

Animals↗