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Biomedical subjects

M Jiang

Publications and source records attributed to M Jiang.

At least 55 records · Page 3Linked to original sources

Quantification of red blood cell fragmentation by automated haematology analyser XE-2100.

The extent of red blood cell fragmentation in peripheral blood is useful for diagnosis and follow-up in many diseases, e.g. haemolytic uremic syndrome, transplantation-associated thrombotic microangiopathy (BMT-TMA). However, this quantification still relies on manual counting of fragmented red cells on blood smears. We have developed a quantification system by gating a fixed area of fragmented red blood cells (Gate 1) on an automated haematology analyser (XE-2100, Sysmex Co., Kobe, Japan). The fragmented red cell percentage (FRC%) calculated with this system, from 100 samples, was highly correlated with the manual count (r=0.902, P < 0.0001). Because microcytic anaemia specimens usually occupy a lower position on the XE-2100 scattergram, with microcytic cells overlapping Gate 1 and causing a spuriously high FRC% calculation, a supplementary gate (Gate 2) was added. Using the particle number in this gate as well as in Gate 1, a revised method for such samples was developed and its validity confirmed (revised FRC% correlated with a manual count for 10 subjects (P < 0.001). Because this gating system can be programmed on any XE-2100, it is likely to prove useful for accurate quantification of red blood cell fragmentation and for the monitoring of the development of BMT-TMA.

Anemia, Iron-Deficiency↗

Effect of abscisic acid on active oxygen species, antioxidative defence system and oxidative damage in leaves of maize seedlings.

Leaves of maize (Zea mays L.) seedlings were supplied with different concentrations of abscisic acid (ABA). Its effects on the levels of superoxide radical (O(2)(-)), hydrogen peroxide (H(2)O(2)) and the content of catalytic Fe, the activities of several antioxidative enzymes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR), the contents of several non-enzymatic antioxidants such as ascorbate (ASC), reduced glutathione (GSH), alpha-tocopherol (alpha-TOC) and carotenoid (CAR), and the degrees of the oxidative damage to the membrane lipids and proteins were examined. Treatment with 10 and 100 microM ABA significantly increased the levels of O(2)(-) and H(2)O(2), followed by an increase in activities of SOD, CAT, APX and GR, and the contents of ASC, GSH, alpha-TOC and CAR in a dose- and time-dependent pattern in leaves of maize seedlings. An oxidative damage expressed as lipid peroxidation, protein oxidation, and plasma membrane leakage did not occur except for a slight increase with 100 microM ABA treatment for 24 h. Treatment with 1,000 microM ABA led to a more abundant generation of O(2)(-) and H(2)O(2) and a significant increase in the content of catalytic Fe, which is critical for H(2)O(2)-dependent hydroxyl radical production. The activities of these antioxidative enzymes and the contents of alpha-TOC and CAR were still maintained at a higher level, but no longer further enhanced when compared with the treatment of 100 microM ABA. The contents of ASC and GSH had no changes in leaves treated with 1,000 microM ABA. These results indicate that treatment with low concentrations of ABA (10 to 100 microM) induced an antioxidative defence response against oxidative damage, but a high concentration of ABA (1,000 microM) induced an excessive generation of AOS and led to an oxidative damage in plant cells.

Abscisic Acid↗

Lack of muscarinic regulation of Ca(2+) channels in G(i2)alpha gene knockout mouse hearts.

The purpose of the present study was to examine the role of G(i2)alpha in Ca(2+) channel regulation using G(i2)alpha gene knockout mouse ventricular myocytes. The whole cell voltage-clamp technique was used to study the effects of the muscarinic agonist carbachol (CCh) and the beta-adrenergic agonist isoproterenol (Iso) on cardiac L-type Ca(2+) currents in both 129Sv wild-type (WT) and G(i2)alpha gene knockout (G(i2)alpha-/-) mice. Perfusion with CCh significantly inhibited the Ca(2+) current in WT cells, and this effect was reversed by adding atropine to the CCh-containing solution. In contrast, CCh did not affect Ca(2+) currents in G(i2)alpha-/- ventricular myocytes. Addition of CCh to Iso-containing solutions attenuated the Iso-stimulated Ca(2+) current in WT cardiomyocytes but not in G(i2)alpha-/- cells. These findings demonstrate that, whereas the Iso-G(s)alpha signal pathway is intact in G(i2)alpha gene knockout mouse hearts, these cells lack the inhibitory regulation of Ca(2+) channels by CCh. Therefore, G(i2)alpha is necessary for the muscarinic regulation of Ca(2+) channels in the mouse heart. Further studies are needed to delineate the possible interaction of G(i) and other cell signaling proteins and to clarify the level of interaction of G protein-coupled regulation of L-type Ca(2+) current in the heart.

Adrenergic beta-Agonists↗

Integration of perspective and disparity cues in surface-orientation-selective neurons of area CIP.

We investigated the effects of linear perspective and binocular disparity, as monocular and binocular depth cues, respectively, on the response of surface-orientation-selective (SOS) neurons in the caudal part of the lateral bank of the intraparietal sulcus (area CIP). During the single-unit recording, monkeys were required to perform the delayed-matching-to-sample (successive same/different discrimination) of discriminating surface orientation in stereoscopic computer graphics. Of 211 visually responsive neurons, 66 were intensively tested using the solid-figure stereogram (SFS) of a square plate with both disparity and perspective cues (D+P condition), and 62 of these were identified as SOS neurons for responding selectively to the orientation of stimuli. All these neurons were further tested using a solid figure with perspective cues alone (P-only condition), and 58% (36/62) of these showed selective response to the orientation of the stimuli. Of the 62 SOS neurons, 35 neurons were also tested using SFS with disparity cues alone (D-only condition) in addition to the D+P and P-only conditions. We classified these 35 neurons into four groups by comparing the response selectivity under the P-only and D-only conditions. More than one-half of these (19/35) were sensitive to both perspective and disparity cues (DP neurons), and nearly one-third (11/35) of these were sensitive to disparity cues alone (D neurons), but a few (2/35) were sensitive to perspective cues alone (P neurons). The remaining (3/35) neurons exhibited orientation selectivity only when both cues were present. In DP neurons, the preferred orientation under the D+P condition was correlated to those under the D-only and P-only conditions, and the response magnitude under the D+P condition was greater than those under the D-only and P-only conditions, suggesting the integration of both cues for the perception of surface orientation. However, in these neurons, the orientation tuning sharpness under the D+P and D-only conditions was higher than that under the P-only condition, suggesting the dominance of disparity cues. After the single-unit recording experiments, muscimol was microinjected into the recording site to temporarily inactivate its function. In all three effective cases out of six microinjection experiments, discrimination of a three-dimensional (3D) surface orientation was impaired when disparity cues alone were present. In only one effective case, when a relatively large amount of muscimol was microinjected, discrimination of a 3D surface orientation was impaired even when both disparity and perspective cues were present. These results suggest that linear perspective is an important cue for representations of a 3D surface of SOS neurons in area CIP, although it is less effective than binocular disparity, and that both of these depth cues may be integrated in area CIP for the perception of surface orientation in depth.

Algorithms↗

Molecular cloning and characterization of a novel human gene (HSPBAP1) from human fetal brain.

Rat PASS1 is a novel protein binding specifically to hsp27 and thus plays a role in regulating stress response in the living cell. Here we report on a human homologue of PASS1, encoded by the gene HSPBAP1. The gene was cloned and identified during a large-scale sequencing analysis of a human fetal brain cDNA library. The human protein shared 80% amino acid identity with rat PASS1. According to bioinformatics analysis, the HSPBAP1 gene is located on chromosome 3q21. RT-PCR analysis indicated that HSPBAP1 was abundant in thymus and pancreas but had a ubiquitously low expression pattern in other human adult tissues except for brain where it was absent.

Amino Acid Sequence↗

Changes of serum selenium in pregnant women with gestational diabetes mellitus.

Gestational diabetes is one of the most common diseases in pregnancy. In the present work, the possible relationship between serum selenium concentration and gestational diabetes was investigated. Blood samples of 234 pregnant women were collected, including 98 subjects with impaired glucose tolerance (IGT), 46 subjects with gestational diabetes mellitus (GDM), and 90 normal pregnant women (NPW). An additional 17 samples of normal women of fertile age (NW) were collected for comparison. The hydride generation atomic fluorescence spectrometry was used for selenium determination. The mean serum selenium levels obtained for each group were 0.0741 +/- 0.0167 mg/L for NPW, 0.0631 +/- 0.0132 mg/L for IGT, 0.0635 +/- 0.0120 mg/L for GDM, and 0.108 +/- 0.0170 mg/L for NW. Serum selenium levels were significantly lower in pregnant woman with IGT (p < 0.001) and GDM (p < 0.001) than in NPW. Furthermore, an inverse correlation between the serum selenium concentration and the gestational period was also observed. Selenium supplementation during gestation for pregnant women, especially with IGT and GDM, should be considered.

Adult↗

[Invasive rhinocerebral aspergillosis occurred during myelosuppressive phase after chemotherapy: a case report and literature review].

OBJECTIVE: To report a case of invasive rhinocerebral aspergillosis (IRA) during myelosuppressive phase after chemotherapy and the treatment outcome. METHODS: A patient with acute non-lymphoblastic leukemia (M(2a)) occurred IRA during myelosuppressive phase after chemotherapy and was treated with amphotericin B (AmB, including AmB-L), garlicin, 5-flucytosine (5-FC), fluconazole, itraconazole, ketoconazole, econazole and griseofulvin with twice of cerebrotomy lesion resections and cerebral decompressions. RESULTS: Though the survival time of this patient is a little longer than the others, she finally died of uncontrolled infection. CONCLUSION: For most of the IRA, there remains no effective treatment. The best treatment is AmB in combination with surgical excision.

Adult↗

[Long-term efficacy of vaccination against hepatitis B in newborns: 13 years' follow-up].

OBJECTIVE: To evaluate long-term efficacy of hepatitis B vaccination in newborns. METHODS: In the past 13 years we followed up the children who had been immunized with hepatitis B vaccine at birth in 1986 every other year. At each follow-up visit, every child was tested for HBsAg, anti-HBs and anti-HBc. RESULTS: HBsAg positive rate of these birth cohort was low (range between 0.46% and 0.98%) during 13 years' follow-up. Long-term efficacy of the vaccination in preventing HBsAg carriage was confirmed. CONCLUSIONS: Thirteen years later no booster was after needed.

Female↗

[Application of video laryngoscope in diagnosis and treatment of laryngeal diseases].

OBJECTIVE: To investigate the value of video laryngoscope in diagnosis of laryngeal diseases. To study the effect of video laryngoscope in treatment of laryngeal diseases. METHOD: Pentax VNL-1530T and Olympus BF-240T video laryngoscope were used in diagnosis of 3628 cases of laryngeal diseases and in treatment of 1221 cases of laryngeal diseases. RESULT: 3628 cases were examined by video laryngoscope, and correct diagnosis was made. 768 cases of vocal cord polyp and 249 cases of vocal cord node were excised, 113 cases of foreign bodies in hypopharynx or larynx were removed. 91 cases of other laryngeal diseases were treated under video laryngoscope. CONCLUSION: Video laryngoscope is a new excellent instrument in diagnosis and treatment of laryngeal disease.

Adolescent↗

The light response of ON bipolar neurons requires G[alpha]o.

ON bipolar neurons in retina detect the glutamate released by rods and cones via metabotropic glutamate receptor 6 (mGluR6), whose cascade is unknown. The trimeric G-protein G(o) might mediate this cascade because it colocalizes with mGluR6. To test this, we studied the retina in mice negative for the alpha subunit of G(o) (Galpha(o)-/-). Retinal layering, key cell types, synaptic structure, and mGluR6 expression were all normal, as was the a-wave of the electroretinogram, which represents the rod and cone photocurrents. However, the b-wave of the electroretinogram, both rod- and cone-driven components, was entirely missing. Because the b-wave represents the massed response of ON bipolar cells, its loss in the Galpha(o) null mouse establishes that the light response of the ON bipolar cell requires G(o). This represents the first function to be defined in vivo for the alpha subunit of the most abundant G-protein of the brain.

Animals↗

Exploitation of a cellulose-binding domain from Neurospora crassa.

After eight decades as a purely research organism, Neurospora crassa is becoming a production system for heterologous peptides. The present work exploits the cbh-1 gene, which encodes a class C cellobiohydrolase (EC 3.2.1.91) and has, at its carboxy-terminus, a domain with homology to other fungal cellulose-binding domains. We describe the construction of two translational fusions of the putative cellulose-binding domain with a reporter gene, which is the catalytic domain of the gla-1 glucoamylase gene of the same species, their transformation back into the organism, and expression of the constructs as cellulose-binding glucoamylase activity. This adds to the developing biotechnology of the organism the potential for enzyme/protein immobilisation.

Journal Article↗

Novel hippocampal interneuronal subtypes identified using transgenic mice that express green fluorescent protein in GABAergic interneurons.

The chief inhibitory neurons of the mammalian brain, GABAergic neurons, are comprised of a myriad of diverse neuronal subtypes. To facilitate the study of these neurons, transgenic mice were generated that express enhanced green fluorescent protein (EGFP) in subpopulations of GABAergic neurons. In one of the resulting transgenic lines, called GIN (GFP-expressing Inhibitory Neurons), EGFP was found to be expressed in a subpopulation of somatostatin-containing GABAergic interneurons in the hippocampus and neocortex. In both live and fixed brain preparations from these mice, detailed microanatomical features of EGFP-expressing interneurons were readily observed. In stratum oriens of the hippocampus, EGFP-expressing interneurons were comprised almost exclusively of oriens/alveus interneurons with lacunosum-moleculare axon arborization (O-LM cells). In the neocortex, the somata of EGFP-expressing interneurons were largely restricted to layers II-IV and upper layer V. In hippocampal area CA1, two previously uncharacterized subtypes of interneurons were identified using the GIN mice: stratum pyramidale interneurons with lacunosum-moleculare axon arborization (P-LM cells) and stratum radiatum interneurons with lacunosum-moleculare axon arborization (R-LM cells). These newly identified interneuronal subtypes appeared to be closely related to O-LM cell, as they selectively innervate stratum lacunosum-moleculare. Whole-cell patch-clamp recordings revealed that these cells were fast-spiking and showed virtually no spike frequency accommodation. The microanatomical features of these cells suggest that they function primarily as "input-biasing" neurons, in that synaptic volleys in stratum radiatum would lead to their activation, which in turn would result in selective suppression of excitatory input from the entorhinal cortex onto CA1 pyramidal cells.

Action Potentials↗

Spine loss and other dendritic abnormalities in epilepsy.

Studies of neurons from human epilepsy tissue and comparable animal models of focal epilepsy have consistently reported a marked decrease in dendritic spine density on hippocampal and neocortical pyramidal cells. Spine loss is often accompanied by focal varicose swellings or beading of dendritic segments. An ongoing excitotoxic injury of dendrites (dendrotoxicity), produced by excessive release of glutamate during seizures, is often assumed to produce these abnormalities. Indeed, application of glutamate receptor agonists to dendrites can produce both spine loss and beading. However, the cellular mechanisms underlying the two processes appear to be different. One recent study suggests NMDA-induced spine loss is produced by Ca2+-mediated alterations of the spine cytoskeleton. In contrast, dendritic beading is not dependent on extracellular Ca2+; instead, it appears to be produced by the movement of Na+ and Cl- intracellularly and an obligate movement of water to maintain osmolarity. A decrease in dendritic spine density was recently reported in a model of recurrent focal seizures in early life. Unlike results from other models, dendritic beading was not observed, and other signs of neuronal injury and death were absent. Thus, additional mechanisms to those of excitotoxicity may produce dendritic spine loss in epileptic tissue. A hypothesis is presented that spine loss can be a product of a partial deafferentation of pyramidal cells, resulting from an activity-dependent pruning of neuronal connectivity induced by recurring seizures. The dendritic abnormalities observed in epilepsy are commonly suggested to be a product and not a cause of epilepsy. However, anatomical remodeling may be accompanied by alterations in molecular expression and targeting of both voltage- and ligand-gated channels in dendrites. It is conceivable that such changes could contribute to the neuronal hyperexcitability of epilepsy.

Animals↗

Delayed rectifier K currents have reduced amplitudes and altered kinetics in myocytes from infarcted canine ventricle.

OBJECTIVE: The rapid (I(Kr)) and slow (I(Ks)) components of delayed rectifier currents play an important role in determining the cardiac action potential configuration. Abnormalities in their function may contribute to arrhythmogenesis under pathological conditions. We studied the effects of myocardial infarction on I(Kr) and I(Ks) in canine ventricular myocytes and their molecular basis. METHODS: Infarct zone myocytes (IZs) were isolated from a thin layer of surviving epicardium overlying an infarct 5 days after a total occlusion of the left anterior descending (LAD) coronary artery. Normal myocytes (NZs) were isolated from the corresponding region of control hearts for comparison. Currents were recorded under the whole-cell patch clamp conditions. RESULTS: Both I(Kr) and I(Ks) current densities were reduced in IZs versus NZs. Kinetic analysis further suggests an acceleration of I(Kr) activation and I(Ks) deactivation. RNase protection assays were used to quantify the mRNA levels of I(Kr) and I(Ks) channel subunits (dERG, dIsK and dKvLQT1) in tissue immediately adjacent to the region where myocytes were isolated. mRNA levels of all three subunits were reduced 2 days after LAD occlusion (by 48+/-9%, 68+/-5%, and 45+/-4% for dERG, dIsK and dKvLQT1, respectively, n=8 each). By day 5, the dKvLQT1 message returned to control while those of dERG and dIsK remained reduced (by 52+/-7% and 76+/-6%, respectively). CONCLUSIONS: The decrease in I(Kr) and I(Ks) amplitudes and changes in their kinetics in infarcted tissue might be due to a decrease in functional channels and/or changes in their subunit composition. Heterogeneous changes in I(Kr) and I(Ks) in infarcted hearts may impact on the effects of varying heart rate or neurohumoral modulation on repolarization.

Analysis of Variance↗

On-demand electrochemical release of DNA from gold surfaces.

Electrochemical quartz crystal microbalance (EQCM) is used to probe the electrochemically triggered release of nucleic acids from gold surfaces to solutions of physiological pH. The immobilization of nonthiolated DNA onto the gold surface is followed by an electrostatic desorption at -1.0 V (vs. Ag/AgCl). Steady-state frequency signals, corresponding to the removal of 261- and 644-ng/cm2 single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA), respectively, were attained within 75 and 330 s. As expected for electrostatic repulsion, the amount released can be manipulated by tuning the potential. The small nonsteady-state frequency signals observed at lower potentials indicate promise for a sustained DNA release. Applicability to gold ultramicroelectrodes (12.5-microm radius) is demonstrated in connection with voltammetric blocking experiments. We expect that such on-demand electrochemical release would be a useful addition to the arsenal of nonviral gene delivery routes.

DNA↗

Multiple histidine kinases regulate entry into stationary phase and sporulation in Bacillus subtilis.

Protein homology studies identified five kinases potentially capable of phosphorylating the Spo0F response regulator and initiating sporulation in Bacillus subtilis. Two of these kinases, KinA and KinB, were known from previous studies to be responsible for sporulation in laboratory media. In vivo studies of the activity of four of the kinases, KinA, KinC, KinD (ykvD) and KinE (ykrQ), using abrB transcription as an indicator of Spo0A approximately P level, revealed that KinC and KinD were responsible for Spo0A approximately P production during the exponential phase of growth in the absence of KinA and KinB. In vitro, all four kinases dephosphorylated Spo0F approximately P with the production of ATP at approximately the same rate, indicating that they possess approximately equal affinity for Spo0F. All the kinases were expressed during growth and early stationary phase, suggesting that the differential activity observed in growth and sporulation results from differential activation by signal ligands unique to each kinase.

Bacillus subtilis↗

Differential processing of propeptide inhibitors of Rap phosphatases in Bacillus subtilis.

In the phosphorelay signal transduction system for sporulation initiation in Bacillus subtilis, the opposing activities of histidine kinases and aspartyl phosphate phosphatases determine the cell's decision whether to continue with vegetative growth or to initiate the differentiation process. Regulated dephosphorylation of the Spo0A and Spo0F response regulators allows a variety of negative signals from physiological processes that are antithetical to sporulation to impact on the activation level of the phosphorelay. Spo0F approximately P is the known target of two related phosphatases, RapA and RapB. In addition to RapA and RapB, a third member of the Rap family of phosphatases, RapE, specifically dephosphorylated the Spo0F approximately P intermediate in response to competence development. RapE phosphatase activity was found to be controlled by a pentapeptide (SRNVT) generated from within the carboxy-terminal domain of the phrE gene product. A synthetic PhrE pentapeptide could (i) complement the sporulation deficiency caused by deregulated RapE activity of a phrE mutant and (ii) inhibit RapE-dependent dephosphorylation of Spo0F approximately P in in vitro experiments. The PhrE pentapeptide did not inhibit the phosphatase activity of RapA and RapB. These results confirm previous conclusions that the specificity for recognition of the target phosphatase is contained within the amino acid sequence of the pentapeptide inhibitor.

Amino Acid Sequence↗

Thyroid hormone-induced overexpression of functional ryanodine receptors in the rabbit heart.

Modifications in the Ca(2+)-uptake and -release functions of the sarcoplasmic reticulum (SR) may be a major component of the mechanisms underlying thyroid state-dependent alterations in heart rate, myocardial contractility, and metabolism. We investigated the influence of hyperthyroid state on the expression and functional properties of the ryanodine receptor (RyR), a major protein in the junctional SR (JSR), which mediates Ca(2+) release to trigger muscle contraction. Experiments were performed using homogenates and JSR vesicles derived from ventricular myocardium of euthyroid and hyperthyroid rabbits. Hyperthyroidism, with attendant cardiac hypertrophy, was induced by the injection of L-thyroxine (200 microg/kg body wt) daily for 7 days. Western blotting analysis using cardiac RyR-specific antibody revealed a significant increase (>50%) in the relative amount of RyR in the hyperthyroid compared with euthyroid rabbits. Ca(2+)-dependent, high-affinity [(3)H]ryanodine binding was also significantly greater ( approximately 40%) in JSR from hyperthyroid rabbits. The Ca(2+ )sensitivity of [(3)H]ryanodine binding and the dissociation constant for [(3)H]ryanodine did not differ significantly between euthyroid and hyperthyroid hearts. Measurement of Ca(2+)-release rates from passively Ca(2+)-preloaded JSR vesicles and assessment of the effect of RyR-Ca(2+)-release channel (CRC) blockade on active Ca(2+)-uptake rates revealed significantly enhanced (>2-fold) CRC activity in the hyperthyroid, compared with euthyroid, JSR. These results demonstrate overexpression of functional RyR in thyroid hormone-induced cardiac hypertrophy. Relative abundance of RyR may be responsible, in part, for the changes in SR Ca(2+) release, cytosolic Ca(2+) transient, and cardiac systolic function associated with thyroid hormone-induced cardiac hypertrophy.

Animals↗