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Biomedical subjects

Q Chen

Publications and source records attributed to Q Chen.

At least 415 records · Page 23Linked to original sources
PubMed1998

Purification and kinetic characterization of hexokinase and glucose-6-phosphate dehydrogenase from Schizosaccharomyces pombe.

Hexokinase and D-glucose-6-phosphate dehydrogenase (G6PDH) from Schizosaccharomyces pmbe have been purified 250-fold by an identical three-step. Both enzymes are dimeric with a molecular mass of 88 kDa for the kinase and 112 kDa for the dehydrogenase. Steady-state kinetic studies were performed on hexokinase and G6PDH, which form the glucose phosphate branch of the oxidative pentose phosphate pathway of S. pombe (fission yeast). Hexokinase promotes Mg(2+)-activated phosphorylation of D-glucose by the equilibrium random Bi Bi mechanism with formation of the abortive enzyme-ADP-glucose complex. ADP inhibits the kinase competitively versus ATP and noncompetitively versus D-glucose. The Mg2+ activation of hexokinase is associated with an increase in the maximal velocity by its interaction with the ternary complex to facilitate the transfer of the phosphoryl group. G6PDH catalyzes NADP(+)-linked oxidation of D-glucose-6-phosphate by the ordered Bi Bi mechanism with NADP+ as the leading reactant. High NADP+ concentration inhibits the dehydrogenase by forming the dead-end ternary complex. In addition, G6PDH is also subjected to product inhibition by NADPH and noncompetitive inhibition by A(G)TP. Thus, the oxidative pentose phosphate pathway in S. pombe may be regulated via inhibition of hexokinase by ADP in conjunction with inhibition of G6PDH by NADPH and ATP.

Glucosephosphate Dehydrogenase↗
PubMed1998

Regulation of D-glucose-6-phosphate dehydrogenase from Schizosaccharomyces pombe.

D-Glucose-6-phosphate dehydrogenase is a regulatory enzyme of the oxidative pentose phosphate pathway in Schizasaccharomyces pombe. The enzyme is subject to negative cooperative regulation by D-glucose-6-phosphate as characterized by the Hill coefficient of 0.68 +/- 0.04. D-Glyceraldehyde-3-phosphate and D-ribulose-5-phosphate rectify the negative cooperativity as evidenced from a change in the Hill coefficients to 0.98 +/- 0.05 and 1.02 +/- 0.05, respectively. These pentose phosphate pathway intermediates also inhibit the enzyme competitively with respect to D-glucose-6-phosphate. Thus, D-glucose-6-phosphate dehydrogenase provides an avenue for regulating the partitioning of D-glucose between the redundant branches of the oxidative phosphate pathway in S. pombe.

Dose-Response Relationship, Drug↗
PubMed1998

Impaired G protein function in gallbladder muscle from progesterone-treated guinea pigs.

This study was designed to elucidate the mechanism of action of progesterone on gallbladder smooth muscle in guinea pigs. Adult male guinea pigs were treated with either progesterone (2 mg.kg-1.day-1) or saline for 7 days. Gallbladder muscle cells were isolated by enzymatic digestion with collagenase. Contractile responses to agonists were expressed as percent shortening from control cell length. [35S]guanosine 5'-O-(3-thiotriphosphate) ([35S]GTP gamma S)-binding properties of G proteins were assessed in crude membranes of gallbladder muscle with or without cholecystokinin octapeptide (CCK-8) stimulation. Gallbladder muscle cells from progesterone-treated guinea pigs exhibited an impaired contractile response to CCK-8, GTP gamma S, or aluminum fluoride but a normal response to potassium chloride or D-myo-inositol 1,4,5-trisphosphate compared with controls. Western blot analysis of gallbladder muscle revealed the presence of Gi1-2, Gi3, Gq/11, and Gs proteins. The maximal contraction induced by CCK-8 was blocked by pertussis toxin and Gi alpha 3-specific antibodies, but not by Gi alpha 1-2 or Gq/11 alpha antibodies. CCK-8 caused a significant increase in [35S]GTP gamma S binding to Gi alpha 3, but not to Gq/11 alpha or Gi alpha 1-2. The stimulation of Gi alpha 3 binding, however, was significantly reduced in gallbladder muscle membranes from progesterone-treated guinea pigs compared with that in control animals. In conclusion, progesterone might cause gallbladder hypomotility by downregulating Gi3 proteins.

Aluminum Compounds↗
PubMed1998

Signal transduction pathways mediating CCK-induced gallbladder muscle contraction.

The signal transduction that mediates CCK-induced contraction of gallbladder muscle was investigated in the cat. Contraction was measured by scanning micrometry in single muscle cells isolated enzymatically with collagenase. Production of D-myo-inositol 1,4, 5-trisphosphate (IP3) and sn-1,2-diacylglycerol (DAG) was quantitated using HPLC and TLC, respectively. Protein kinase C (PKC) activity was determined by measuring the phosphorylation of a specific substrate peptide from myelin basic protein, Ac-MBP-(4-14). CCK-induced contraction was blocked by incubation in strontium medium, pertussis toxin (PTx), and antibodies against Gialpha3 or betagamma-subunits but was not blocked by Ca2+-free medium or by antibodies against Gq/11alpha, Gialpha1-2, or Goalpha. The contraction induced by CCK was inhibited by the phospholipase C (PLC) inhibitor U-73122, anti-PLC-beta3 antibody, and the IP3 receptor antagonist heparin but was not inhibited by the the phospholipase D inhibitor propranolol or antibodies against PLC-beta1 or PLC-beta2. Western blot analysis of gallbladder muscle revealed the presence of PLC-beta2 and PLC-beta3 but not PLC-beta1. CCK caused a 94% increase in IP3 generation and an 86% increase in DAG generation. A low dose of CCK caused PKC translocation, and CCK-induced contraction was blocked by the PKC inhibitor H-7. A high dose of CCK, however, caused no PKC translocation, and its contraction was blocked by the calmodulin antagonist CGS9343B. In conclusion, CCK contracts cat gallbladder muscle by stimulating PTx-sensitive Gi 3 protein coupled with PLC-beta3, producing IP3 and DAG. Low doses activate PKC, whereas high doses activate calmodulin.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1998

Mechanism for secretagogue-induced surfactant protein A binding to lung epithelial cells.

Secretagogues stimulate both secretion and reuptake of surfactant components by pulmonary type II cells as well as enhance surfactant protein A (SP-A) binding. We have evaluated the possibility that the observed increase in SP-A binding is due to the movement of SP-A receptors from an intracellular pool to the plasma membrane. We utilized an anti-idiotypic monoclonal antibody, A2R, which recognizes an SP-A binding protein on type II cell membranes. Immunocytochemistry studies showed that A2R reacted with cellular antigens on type II cell membranes and paranuclear granules. A2R inhibited cell association of 125I-SP-A to type II cells plated on Transwell membranes as well as those plated on plastic dishes and also inhibited the SP-A-stimulated incorporation of phosphatidylcholine liposomes into type II cells. On exposure to secretagogues, the binding of 125I-A2R and 125I-SP-A to type II cells increased in parallel. With permeabilized type II cells on Transwell membranes, one-sixth of the binding sites were located on the plasma membrane, with the remainder being intracellular; phorbol 12-myristate 13-acetate treatment increased the binding of A2R to the cell surface but did not affect the total binding of A2R. Ligand blots of type II cell plasma membranes showed that SP-A and A2R both bound proteins with molecular masses of approximately 32 and 60 kDa, respectively, reduced. Under nonreducing conditions, the mass of the SP-A and A2R binding protein was approximately 210 kDa, indicating that the SP-A receptor is composed of disulfide-linked subunits. The results support our hypothesis that secretagogues increase SP-A binding sites by accelerating recruitment of receptors to the cell surface.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1998

Comparative changes in uterine artery blood flow waveforms in singleton and twin pregnancies.

We prospectively studied 99 women with uncomplicated singleton pregnancies and 24 women with uncomplicated twin pregnancies to examine the effect of the location of the placenta on the pulsatility index (PI) in the uterine arteries in twin pregnancies and to compare singleton and twin pregnancies with regard to changes in the PI of uterine arteries over the course of the pregnancy. The flow velocity waveforms in both the right and left parametrial uterine arteries were recorded longitudinally at 2-week intervals from a gestational age of 19 weeks until delivery. The location of the placenta was determined by real-time ultrasonography at 18-20 weeks of gestation to determine which uterine artery was the ipsilateral artery. In both the ipsilateral and contralateral uterine arteries, the PI exhibited a steady decrease toward term in singleton pregnancies. In twin pregnancies, the PI exhibited a steady decrease until 27 weeks of gestation and remained unchanged thereafter. The PI in the ipsilateral artery was consistently lower than that in the contralateral artery in singleton as well as twin pregnancies. The mean values of PI of the ipsilateral and contralateral uterine arteries of twin pregnancies was consistently lower than that of singleton pregnancies at any gestational week. These results suggest that the ipsilateral uterine artery contributes more to placental perfusion than does the contralateral uterine artery in twin as well as in singleton pregnancies. Twin pregnancies differed from singleton pregnancies with respect to changes in PI in the uterine artery according to gestational age. The PI of uterine arteries declined more rapidly and reached a plateau earlier in twin than in singleton pregnancies.

Adult↗
PubMed1998

Physiological and molecular properties of AMPA/Kainate receptors expressed by striatal medium spiny neurons.

The mechanisms by which glutamate shapes the activity of striatal medium spiny neurons are of fundamental importance to our understanding of normative and pathological striatal physiology. Non-N-Methyl-D-aspartate (non-NMDA) glutamate receptor expression and function were studied in medium spiny neurons with a combination of single cell RT-PCR, immunocytochemistry and whole-cell voltage-clamp techniques. Reverse transcription polymerase chain reaction analysis found that GluR2 mRNA appeared to be the most abundant and widely distributed AMPA receptor mRNA. GluR1 was also commonly detected. However, GluR3 mRNA was preferentially expressed by neurons coexpressing substance P and enkephalin and GluR4 mRNA was not detected in identified medium spiny neurons. All neuronal classes appeared to express GluR5 or GluR6 and/or GluR7 mRNA in addition to kainate (KA) subunit mRNA. Immunocytochemical studies confirmed the mRNA distributions and also revealed that GluR1 protein was largely restricted to dendritic spines. Although the mRNA and protein for both alpha-amino-3-hydroxy-5-methyl-ioxyzole-4-proprionic acid (AMPA) and KA class subunits was detected, the physiological response to glutamatergic ligands and the benzothiadizine cyclothiazide was characteristic of AMPA, not KA receptors. The AMPA receptor antagonist GYKI 52466 blocked the response to AMPA and all but a small transient component of the response to KA. The current-voltage relationship of the AMPA-evoked currents was relatively linear but Ca2+ fluorometry revealed that substantial changes in intracellular Ca2+ concentration accompanied exposure to either agonist. These results argue that somatodendritic non-NMDA glutamate receptors in medium spiny neurons are primarily GluR2-containing receptors of the AMPA class but that activation of these receptors as a group nevertheless results in a significant Ca2+ influx.

Animals↗
PubMed1998

Genetics, molecular mechanisms and management of long QT syndrome.

Cardiac arrhythmias cause more than 300,000 sudden deaths each year in the USA alone. Long QT syndrome (LQT) is a cardiac disorder that causes sudden death from ventricular tachyarrhythmias, specifically torsade de pointes. Four LQT genes have been identified: KVLQT1 (LQT1) on chromosome 11p15.5, HERG (LQT2) on chromosome 7q35-36, SCN5A (LQT3) on chromosome 3p21-24, and MinK (LQT5) on chromosome 21q22. SCN5A encodes the cardiac sodium channel, and LQT-causing mutations in SCN5A lead to the generation of a late phase of inactivation-resistant whole-cell inward currents. Mexiletine, a sodium channel blocker, is effective in shortening the QT interval corrected for heart rate (QTc) of patients with SCN5A mutations. HERG encodes the cardiac I(Kr) potassium channel. Mutations in HERG act by a dominant-negative mechanism or by a loss-of-function mechanism. Raising the serum potassium concentration can increase outward HERG potassium current and is effective in shortening the QTc of patients with HERG mutations. KVLQT1 is a cardiac potassium channel protein that interacts with another small potassium channel MinK to form the cardiac I(Ks) potassium channel. Like HERG mutations, mutations in KVLQT1 and MinK can act by a dominant-negative mechanism or a loss-of-function mechanism. An effective treatment for LQT patients with KVLQT1 or MinK mutations is expected to be developed based on the functional characterization of the I(Ks) potassium channel. Genetic testing is now available for some patients with LQT.

Chromosome Mapping↗
PubMed1998

Purification and kinetic characterization of 6-phosphogluconate dehydrogenase from Schizosaccharomyces pombe.

6-Phosphogluconate dehydrogenase is the pivotal enzyme that links the gluconate route and the oxidative phase of the pentose phosphate pathway in Schizosaccharomyces pombe. The enzyme differs from the known 6-phosphogluconate dehydrogenases of other sources in that the Schizosaccharomyces enzyme is tetrameric having a subunit mass of 38 kDa, that it requires NADP+ obligatorily for activity, and that it can be activated by divalent metal ions such as Co2+ and Mn2+. Steady-state kinetic studies were undertaken. Initial rate and product inhibition results suggest that 6-phosphogluconate dehydrogenase from Schizosaccharomyces pombe catalyzes NADP(+)-linked oxidative decarboxylation of 6-phosphogluconate by an equilibrium random mechanism with two independent binding sites, namely one site for the nicotinamide coenzyme, NADP+/NADPH, and another site for 6-phosphogluconate-D-ribulose-5-phosphate and for CO2. Studies of pH dependence implicated a basic residue with a pK value of 7.4 in the binding of 6-phosphogluconate and an acidic residue with a pK value of 6.7 in the cation-mediated interaction of NADP+ with the enzyme.

Decarboxylation↗
PubMed1998

[Application of biomarkers in homologous clone identification of enterotoxigenic Escherichia coli].

OBJECTIVE: To study the application of multiple molecular biotechniques and multi-biomarkers in homologous clone identification of enterotoxigenic Escherichia coli (ETEC). METHODS: On the basis of detection and typing for toxigenic genes, plasmid analysis combined with multilocus enzyme electrophoresis (MEE) was used for the first time in the comprehensive research on ETEC homologous clone. RESULTS: Two hundred and six strains of ETEC could be classified into 93 plasmid types with plasmid profiling, and 140 strains with unique genotype. Those 206 strains of ETEC could be classified into 148 electrophoresis-types by MEE further, and altogether 190 strains with unique genotype, by combination of three methods. CONCLUSION: Multiple molecular biotechniques and multi-biomarkers can provide much more and more reliable information in homologous clone identification.

Diarrhea↗
PubMed1998

[Establishment of transgenic cell line CHL-3A4 and its metabolic activation].

OBJECTIVE: To establish a model cell line CHL-3A4 to study metabolic pathways for some genotoxic chemicals. METHODS: Complimentary DNA (cDNA) of cloned cytochrome P450-3A4 gene in human liver tissue was transferred to Bluescript M13 vector with reverse transcription polymerase chain reaction (RT-PCR) and DNA recombinant techniques. Restriction endonuclease map analysis and sequencing of partial cloned fragment proved that CYP3A4 cDNA was cloned into Bluescript M13 vector. Then, recombinant expression plasmid pREP9-3A4 was constructed in eukaryotic cell and transfected into Chinese hamster CHL cells. RESULTS: It was proved that a CHL-3A4 transgenic cell line was established, which could lead metabolic activation for aflatoxin B1 (AFB1), sterigmatocystin (STC) and cyclophosphamide (CPA). CONCLUSION: The CHL-3A4 cell line established did express human cytochrome P450 3A4 and could lead metabolic activation for three genotoxic chemicals AFB1, STC and CPA.

Animals↗
PubMed1998

[An in vitro study on toxic effect of vanadium-titanium-magnetite dust on alveolar macrophage in rabbits].

OBJECTIVE: To study the toxic effect of vanadium-titanium-magnetite (VTM) dust on alveolar macrophage (AM) and its hazardous extent. METHODS: Survival rates, morphology and function of AM were compared in rabbits exposed to dust of VTM, vanadium oxide, titanium dioxide and silica in various doses and length of time with in vitro cell culture and putamen membrane cover glass transmission electron microscopy, and changes in activities of lactic dehydrogenase (LDH) and acid phosphatase (ACP) in cell culture were measured. RESULTS: Exposure to all the four kinds of dust could lead to decrease in survival rate of AM, increase in activities of LDH and ACP in the cell culture, and changes in their morphology and function to the extent dependent on the nature of dust. CONCLUSION: Toxic effect of exposure to VTM dust was lower than that to vanadium oxide and silica, but higher than that to titanium dioxide, which had slight toxic effect.

Acid Phosphatase↗
PubMed1998

[Changes of tissue oxygenation and relationship between oxygen delivery and consumption in patients with poor cardiac function undergoing valve replacement].

To study the relationship between oxygen delivery(DO2) and consumption(VO2) and tissue oxygenation in patients during cardiac valve replacement, DO2, VO2, arterial blood lactate (ABL) SvO2 were measured. The results showed a poor correlation between DO2 and VO2 and a normal tissue oxygenation before CPB. There was a good positive correlation between DO2 and VO2 within 20 h after the end of CPB(P < 0.01), and changes of ABL and SvO2 showed tissue hypoxia. The level of ABL in patients showing O2 supply-dependency was lower, and SvO2 was higher than those in 15 patients showing O2 supply-independency, respectively (P < 0.05). These data suggest that O2 supply-dependency was existed after the end of CPB, and there was a good correlation between O2 supply-dependency and ABL. It might not exclude tissue hypoxia in O2 supply-independency and normal level of SvO2.

Adult↗
PubMed1998

[The morphometric study of endometrial spiral arterioles before and after insertion of gamma CuI and TCu 220C intrauterine devices].

OBJECTIVE: To investigate the morphologic changes of endometrial spiral arterioles and its relationship with bleeding pattern after insertion of gamma-shape copper indomethacin-medicated (gamma CuI) and T-shape copper (TCu 220C) intrauterine devices (IUD). METHODS: Endometrium specimens of late secretory phase were obtained from fertile age women: 10 from preinsertion, 10 obtained after insertion of TCu 220C IUD, and 9 obtained after insertion of gamma CuI IUD. Samples were sectioned serially and morphometric analysis of endometrial spiral arterioles was performed under light microscope. RESULTS: The average cross section area (Area), maximum diameter (Dmax) and minimum diameter (Dmin) of spiral arterioles in both spongeous and dense layers of endometrium increased significantly after insertion of TCu 220C IUD. After insertion of gamma CuI IUD, the Area and Dmax increased in dense layer only, though less obviously than that occurred in TCu 220C group. However, the Dmin increased more obviously in both spongeous and dense layers than after insertion of TCu 220C IUD, implying that the shape of spiral arterioles was more regular in gamma CuI group. CONCLUSION: gamma CuI IUD has less effects on the morphological changes of endometrial spiral arterioles, and this may relate to its indomethacin-contained which causes less bleeding.

Adult↗