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Biomedical subjects

G Luo

Publications and source records attributed to G Luo.

At least 145 records · Page 8Linked to original sources

[The role of thermal injury on nitric oxide (NO) production by mouse macrophages and its possible mechanism].

To study the production NO by macrophages (M phi) after burn injury and the possible mechanism, we determined the NO production and the iNOS (inducible NO synthase) activity of peritoneal M phi (PM phi) in scald mouse, and the effect of iNOS specific blocker NMMA. Actinomycin D (AD) and protein tyrosine kinase (PTK), special inhibitor Genisteinon them in vitro. It was found that PM phi produced excessive NO in the early postburn phase and the iNOS activity was increased significantly, and there was a positive correlation between them. NMMA, AD as well as Genistein could inhibit the NO production, the activity of iNOS was decreased by both AD and Genistein. The results suggested that M phi was activated after thermal injury and it could produce NO excessively through initiating iNOS synthesis. PTK signal system was involved in the NO production by M phi via affecting the iNOS synthesis after burn injury.

Animals↗

[The study of local electroretinogram in age-related macular degeneration].

PURPOSE: Observing the features of electro-activities at macular region in age-related macular degeneration (AMD), and evaluating the sensitivities of local electroretinogram (LERG). METHODS: The LERG of 2.5, 5, 7.5 degrees at macula had been recorded in 99 eyes (68 dry form, 31 wet form) of 80 AMD patients. The contrast study had also been carried out with visual acuities and lesion areas at the corresponding region. RESULTS: The results showed that there were no significant differences between the decreased amplitudes of a and b waves of LERG in dry form AMD with normal controls (P > 0. 05). While significant differences existed in wet form AMD (P < 0.01). The changes of LERG at 2.5 degrees were greater than those of the other two regions. The correlation study on b wave amplitude and visual acuity of AMD patients showed that there was a positive correlation existed between them only in wet form AMD at 2.5 degrees region (r = 0.516, P < 0.05). The negative correlations were observed between amplitudes of b wave and lesion areas in wet form AMD at the three macular regions. Drusen of dry form AMD had no effects on the amplitudes of LERG. CONCLUSIONS: The impairments on macular function from two types of AMD are different. This study reveals that the LERG is a sensitive method of visual electrophysiology for measuring macular function. The smaller the testing area, the higher the sensitivity.

Aged↗

[Analysis of cinnamic acid in storax and its original plant by HPLC].

The contents of free and hydrolytic cinnamic acid in storax and its original plant (Liquidambar orientalis) were determined by high performance liquid chromatography, and the chromatographic behaviors were studied. In styrax gummi, the contents of free and hydrolytic cinnamic acid were 7.03% and 25.26% respectively; and in its original plant, they were 0.3% evenly.

Chromatography, High Pressure Liquid↗

Human neoplasms elicit multiple specific immune responses in the autologous host.

Expression of cDNA libraries from human melanoma, renal cancer, astrocytoma, and Hodgkin disease in Escherichia coli and screening for clones reactive with high-titer IgG antibodies in autologous patient serum lead to the discovery of at least four antigens with a restricted expression pattern in each tumor. Besides antigens known to elicit T-cell responses, such as MAGE-1 and tyrosinase, numerous additional antigens that were overexpressed or specifically expressed in tumors of the same type were identified. Sequence analyses suggest that many of these molecules, besides being the target of a specific immune response, might be of relevance for tumor growth. Antibodies to a given antigen were usually confined to patients with the same tumor type. The unexpected frequency of human tumor antigens, which can be readily defined at the molecular level by the serological analysis of autologous tumor cDNA expression cloning, indicates that human neoplasms elicit multiple specific immune responses in the autologous host and provides diagnostic and therapeutic approaches to human cancer.

Antibodies, Neoplasm↗

Cell-free synthesis of polyketides by recombinant erythromycin polyketide synthases.

Modular polyketide synthases (PKSs) are complex multi-enzyme proteins that catalyse the bacterial biosynthesis of many pharmaceutically useful polyketides. The PKSs are organized into a series of modules, each containing the active catalytic sites required for one step in the synthesis process. Here we report a method for cell-free enzymatic synthesis of 6-deoxyerythronolide B (6-dEB), the parent molecule of the antibiotic erythromycin A, using recombinant 6-deoxyerythronolide B synthase (DEBS), a modular PKS with at least 28 distinct active sites. We have also synthesized in vitro a triketide lactone by using a truncated mutant of DEBS. The availability of such cell-free synthetic routes will allow direct investigation of the structural and mechanistic basis for the unusual combination of high substrate specificity and tolerance to genetic reprogramming found in this enzyme family.

Amino Acid Sequence↗

BMP-7 is an inducer of nephrogenesis, and is also required for eye development and skeletal patterning.

Bone morphogenetic proteins (BMPs) are multifunctional growth factors originally identified by their ability to induce ectopic bone formation. To investigate the function of one of the BMPs, BMP-7, we have generated BMP-7-deficient mice using embryonic stem cell technology. BMP-7-deficient mice die shortly after birth because of poor kidney development. Histological analysis of mutant embryos at several stages of development revealed that metanephric mesenchymal cells fail to differentiate, resulting in a virtual absence of glomerulus in newborn kidneys. In situ hybridization analysis showed that the absence of BMP-7 affects the expression of molecular markers of nephrogenesis, such as Pax-2 and Wnt-4 between 12.5 and 14.5 days postcoitum (dpc). This identifies BMP-7 as an inducer of nephrogenesis. In addition, BMP-7-deficient mice have eye defects that appear to originate during lens induction. Finally, BMP-7-deficient mice also have skeletal patterning defects restricted to the rib cage, the skull, and the hindlimbs.

Animals↗

The matrix Gla protein gene is a marker of the chondrogenesis cell lineage during mouse development.

Matrix Gla protein (MGP) is, along with osteocalcin, a skeletal member of the family of extracellular mineral-binding Gla proteins. Although the precise function of these proteins remains obscure, circumstantial evidence suggests that they play a role in endochondral ossification. As a first step toward understanding MGP function we have performed a preliminary characterization of its promoter element and studied the developmental pattern of expression of this gene. DNA transfection experiments indicate that the mouse MGP promoter functions better in cells expressing the MGP gene than in cells that do not express the gene. During mouse development, MGP gene expression is detectable as early as day 10.5 of embryonic development (E10.5), before any skeletal structures are identifiable. In situ hybridization analysis shows that MGP mRNA is initially present at the mesenchymal epithelial interphase in lung and limb buds. As development proceeds, MGP gene is predominantly expressed in cells of the chondrocytic lineage in areas that will undergo endochondral ossification as well as in areas that will remain cartilaginous, such as the trachea and bronchi. In growth plate cartilage, MGP mRNA is present in resting, proliferative, and late hypertrophic chondrocytes. Surprisingly, MGP mRNA is absent from the early hypertrophic chondrocytes and from the osteoblasts. Finally, the MGP gene is expressed at a lower level in kidney medulla and uterus smooth muscle but not in brain, spleen, or heart during development. This study demonstrates that during development MGP gene expression occurs early and is predominant at the epithelial mesenchymal interfaces, principally of lung and limb buds, and in cells of the chondrocytic lineage. This finding raises the intriguing possibility that MGP may play distinct roles during embryogenesis and in the adult organism.

Animals↗

Implementation of strain rate as a bone remodeling stimulus.

Strain rate is implemented as a stimulus for surface bone remodeling. Using idealized models for trabecular bone structures, the surface remodeling predictions using the strain rate as the stimulus are compared with the predictions using the peak strain magnitude as the stimulus. For a uniaxially loaded cruciform shape, the comparison shows that the two surface remodeling stimuli predict the same final shape under a periodic compressive load, but the two evolutionary paths to final shapes are different. Two biaxially loaded regular grid models of trabecular structure were considered, one a grid of square diamond shaped elements and the other a brick wall patterned grid. For both of these idealized trabecular structures, the comparison shows that the two surface remodeling stimuli predict the same final shape under a periodic compressive load, even from these distinctly different initial grid patterns, and the evolutionary paths to final shapes are quite different. In general the two stimuli do not predict the same remodeling and the conditions under which they do are derived. The models developed are also applied to the data from the animal experiments reported in Goldstein et al. (1991), and it is shown that the strain rate stimulus predicts bone remodeling similar to what was experimentally observed.

Animals↗

Growth impairment resulting from expression of influenza virus M2 protein in Saccharomyces cerevisiae: identification of a novel inhibitor of influenza virus.

The gene encoding M2, the ion channel-forming protein of influenza virus A, was expressed under the control of an inducible promoter in Saccharomyces cerevisiae. By using single and multicopy plasmids containing GAL promoter-M2 fusions, a correlation was observed between plasmid copy number and growth in medium inducing M2 expression. Cells expressing M2 from multicopy plasmids have reduced growth rates, suggesting that high levels of M2 are toxic to growth. The addition of amantadine, a compound known to block the ion channel activity of certain M2 alleles, restores the growth rates to wild-type levels in cells expressing an amantadine-susceptible allele of M2 but not an amantadine-resistant allele of M2, suggesting that M2 expression in S. cerevisiae results in the formation of functional M2 ion channels. Measurements of extracellular acidification by microphysiometry suggest that proton efflux in M2-expressing cells is altered and that the addition of amantadine permits the reestablishment of the proton gradient. The growth impairment phenotype resulting from M2 expression was used to develop a high-capacity screening assay which identified a novel inhibitor possessing an antiviral profile similar to that of amantadine.

Amantadine↗

Erythromycin biosynthesis. Highly efficient incorporation of polyketide chain elongation intermediates into 6-deoxyerythronolide B in an engineered Streptomyces host.

Feeding of (2S,3R)-[2,3-13C2]-2-methyl-3-hydroxypentanoyl NAC thioester (1a) to the recombinant organism Streptomyces coelicolor CH999/pCK7 harboring the complete set of eryA genes from Saccharopolyspora erythraea encoding the 6-deoxyerythronolide B synthase (DEBS) resulted in the formation of 6-deoxyerythronolide B (2a) labeled with 13C at C-12 and C-13, as evidenced by the appearance of a pair of enhanced and coupled doublets in the 13C NMR spectrum. The level of 13C enrichment was 15-20 atom% 13C, as much as 100 times higher than the usually observed efficiency of incorporation of NAC thioesters into polyketide metabolites. Similar incorporation of (2S,3R)-[3-2H,3-13C]-2-methyl-3-hydroxypentanoyl NAC thioester (1b) gave 6-deoxyerythronolide B (2b) labeled with both 13C and deuterium at C-13. The intact incorporation of both precursors confirms the normal functioning of the recombinant DEBS proteins in the heterologous host.

DNA, Recombinant↗

Observation and quantitation of exocytosis from the cell body of a fully developed neuron in Planorbis corneus.

We have discovered a neuronal system that releases neurotransmitter via exocytosis from the cell body. In the large dopamine cell of the pond snail Planorbis corneus, depolarization induces rhythmic release of dopamine from the cell body. When a stimulant is applied extracellularly or intracellularly in situ to the cell body, transient dopamine concentration packets that appear in a bursting pattern are observed. Dopamine release is calcium dependent and release is on the time scale expected for exocytosis (2 to 4 msec rise times). Quantitation of individual events reveals an average of 818,000 molecules per exocytotic event. As many as 89,000 individual exocytotic events have been observed following a single stimulation of one cell. Neurotransmitter exocytosis from the neuronal cell body appears to represent an alternative form of neurocommunication to synaptic transmission.

Animals↗

[Determinate titer of tuberculosis antibody with cytomembrane antigen].

Using tuberculous cytomembrane antibody and titer determining methodology, serum specimens from 165 pulmonary tuberculosis patients were analysed. The results showed: 75.7% of specimen is in high level, titer > or = 1:400.20% in low level, titer 1:100-1:200.4. 42% negative. This method is more sensitive than usually used PPD and polymeric OT antigen. At the same time, sputum specimens from 406 TB patients were examined for TB bacilli, the result demonstrated that the positive rate of sputum smear in the patients with titer > or = 800 is up to 95.71%-100%.1:400 is 69.86%, 1:100-1:200 is 14%-22.09%, < 1:100 is 10%.

Antibodies, Bacterial↗

Metabolism of allylbenzene 2',3'-oxide and estragole 2',3'-oxide in the isolated perfused rat liver.

The metabolism of allylbenzene 2',3'-oxide, estragole 2',3'-oxide, allylbenzene and estragole was studied in the isolated perfused rat liver. Formation of dihydrodiol and glutathione conjugate metabolites was detected for both epoxides and the presence of dihydrodiol metabolites after perfusion of allylbenzene or estragole indicated the formation of allylic epoxide intermediates in the intact liver. A comparison of elimination kinetics for parent compounds and epoxides indicated that epoxides were relatively rapidly detoxified and probably do not accumulate on formation in vivo. Acute toxicity of epoxides, measured as the release of alanine aminotransferase activity into the perfusate, or genetic toxicity, determined as covalent binding of radiolabeled epoxide to DNA, were not observed. It was concluded that both epoxide hydrolases and glutathione S-transferases can effectively detoxify the allylic epoxides derived from either allylbenzene or estragole and effectively prevent cellular or genetic toxicity of these reactive intermediates. Epoxide hydrolases appear to play the major role in the detoxication of these epoxides in vivo.

Allyl Compounds↗

[Experimental study of implantation of autologous vascular endothelial cells on dacron grafts].

Ten mongrel dogs underwent carotid artery bypass 17 times with dacron grafts, 4mm in inner diameter and 5cm in length. The grafts in experimental group A were immediately seeded with autologous endothelial cells harvested by 0.25% trypsin solution (n = 7). The grafts in experimental group B were seeded with autologous endothelial cells cryopreserved in liquid nitrogen for 1 week (n = 3). The grafts in contrast group was not seeded with any endothelial cells implanted in the contralateral carotid arteries in group A (n = 7). The number of endothelial cells harvested in the two groups was 0.9 +/- 0.3 x 10. Factor VIII related antigen stain method confirmed the cells harvested by 0.25% trypsin solution to be endothelial cells. The grafts were removed and studied at the end of 2, 4, and 6 week after implantation. The total patency rate of group A was 85.7% (6/7), and that of the contrast group 57% (4/7). The weight of thrombus in group A was 40.6 +/- 36.9mg and the contrast group 85.9 +/- 26.3mg (P < 0.01). Scanning electron microscopy revealed that the cells at the middle segment of the grafts in the group A and B had the characteristics of ellipse, and tight intercellular junction, while the middle segment grafts in contrast group were covered by platelets, erythrocytes, leukocytes and fibrin. Transmission electron microscopy, factor VIII related antigen stain method, and vimentin stain method demonstrated that the cells in the group A and B showed characteristics of endothelial cells. It was concluded that endothelialization of the dacron grafts could be accelerated and the quality could be improved by immediate seeding with endothelial cells or by seeding with endothelial cells cryopreserved in liquid nitrogen for 1 week.

Animals↗

[The change of filtration-index of erythrocyte malonyldialdehyde and superoxide dismutase in patients with pregnancy induced hypertension].

OBJECTIVE: To observe the relationship between the filtration-index of erythrocytes and concentration of erythrocytic superoxide dismutase (SOD), plasma malonyldialdehyde (MDA) in cases of pregnancy induced hypertension (PIH). METHODS: We have measured the level of MDA and filtration-index, SOD content of erythrocytes in maternal and cord blood of 31 cases with PIH (group A) and 25 cases of normal pregnancy (group B) using TBA method, nucleopore membraned cell deformability meter and pyrogallol self-oxidizing method. RESULTS: The mean filtration-index level and erythrocytic SOD were significantly higher in group A than those of group B (P < 0.01 and P < 0.05). The mean content of plasmatic MDA in group A increased markedly (P < 0.01) compared to that of group B. There was no difference of these parameters in cord blood between group A and group B. CONCLUSION: The decrease of red cell deformability due to the increase of plasma MDA level may be the underlying mechanism of PIH.

Adult↗

Amperometric monitoring of stimulated catecholamine release from rat pheochromocytoma (PC12) cells at the zeptomole level.

Catecholamine release from rat pheochromocytoma (PC12) cells has been observed at zeptomole levels using dc-amperometric detection with carbon fiber microelectrodes. Time-resolved individual exocytic events from PC12 cells have been recorded and analyzed with 1.2 ms time resolution. The average area under 1912 current transients from 13 PC12 cells corresponds to 190 zmol (114,300 molecules per release event). The average width at half-height of these current transients is 9.3 ms, in agreement with the time frame of exocytosis. The detection limit of the method reported here is as low as 31 zmol. This is the first report of direct electrochemical observation of quantal release from PC12 cells. The successful application of this electrochemical scheme to monitor catecholamines released from small vesicles also suggests that it may be possible to apply this technique to monitor quantal release from synaptic vesicles.

Animals↗

Purification and characterization of Yersinia enterocolitica envelope proteins which induce antibodies that react with human thyrotropin receptor.

Graves' disease is an autoimmune disease mediated by autoantibodies to the thyrotropin receptor (TSHR). Several studies have suggested that the development of Graves' disease may be linked to infection with the enteric pathogen Yersinia enterocolitica. Using the purified recombinant extracellular domain of human TSHR (ETSHR), we have recently shown that immunization of mice with Y. enterocolitica results in the production of antibodies capable of reacting with the ETSHR. In this study, we identify two low molecular weight (5.5 kDa and 8 kDa) envelope proteins of Yersinia containing epitopes that are crossreactive with the TSHR. Identification of these crossreactive envelope proteins was achieved by Western blotting using affinity-purified anti-Y. enterocolitica antibodies that specifically react with the TSHR and, conversely, for envelope proteins of Yersinia. Confirmation that these Yersinia proteins contained crossreactive epitopes with the ETSHR was obtained by immunizing mice with partially purified envelope proteins, which resulted in the production of Abs that recognized the ETSHR. Further, some of the cross-reactive envelope proteins were purified with SDS-PAGE and HPLC. The crossreactive envelope proteins were shown to be chromosomally encoded, exposed on the surface of bacteria, and produced by virulent as well as avirulent strains of Yersinia (Y. pestis, Y. pseudotuberculosis, Y. enterocolitica VW+, and Y. enterocolitica VW-). These results identify for the first time the Yersinia envelope proteins that are crossreactive with the ETSHR. Availability of these proteins will allow future studies to determine whether patients with Graves' disease have a unique immune response against these proteins when compared with healthy individuals.

Animals↗