Biomedical subjects

H Wang

Publications and source records attributed to H Wang.

At least 235 records · Page 13Linked to original sources

PubMed2002

Evaluation of apolipoprotein A-II as a positional candidate gene for familial Type II diabetes, altered lipid concentrations, and insulin resistance.

AIMS/HYPOTHESIS: We hypothesized that apolipoprotein A-II sequence variation was responsible for the observed linkage of Type II (non-insulin-dependent) diabetes mellitus to the apolipoprotein A-II region in Northern European families ascertained for multiple diabetic siblings, and might also influence insulin sensitivity and secretion, non-esterified fatty acids, and lipids. METHODS: We recruited 698 members of 63 families for pedigree studies and additional unrelated people providing 117 diabetic and 130 control subjects. We screened the apolipoprotein A-II gene by single strand conformation polymorphism analysis and fluorescent sequence analysis. Variants were typed by oligonucleotide ligation assay, restriction digest of amplification products, or radioactive fragment analysis for the microsatellite polymorphism. Association of each variant with Type II diabetes was tested in the case-control population by chi-square analysis, or using transmission disequilibrium test in families. Haplotypes were established in families using SIMWALK and tested for association with diabetes and quantitative traits. RESULTS: No detected variant altered the coding sequence of the gene. Three single nucleotide polymorphisms showed modest evidence for an association, but no variant or haplotype was associated with diabetes in families. Similarly, we found no association with non-esterified fatty acid concentrations, HDL concentrations, or fasting insulin. In contrast, we found evidence for an association of some haplotypes and individual variants with 2-h post-challenge glucose and measures of insulin secretion. CONCLUSION/INTERPRETATION: Apolipoprotein A-II is not likely to explain the observed linkage of Type II diabetes, but variation in this gene could alter insulin secretion and post-challenge glucose.

Apolipoprotein A-II↗

PubMed2002

Variability analyses suggest that supraspino-spinal interactions provide dynamic stability in motor control.

Effects of supraspino-spinal feedforward-feedback (FF-FB) interactions on variability in locomotor rhythm and coordination were examined in in vitro brain-spinal cord lamprey preparations. Spinal locomotor networks were activated by applying 0.2 mM N-methyl-DL-aspartate (NMA) to three spinal pools: gill, rostral and caudal. Bathing the brain with zero Ca(2+) saline altered supraspinal-spinal drive and FF-FB interaction while spino-supraspinal feedback was changed by applying NMA to the caudal pool only. Wavelet analyses indicated a non-uniform energy distribution in ventral root (VR) activity that shifted between frequency bands on FF-FB interruption. Wavelet analysis was used to extract 300-s long epochs of low frequency burst rhythm. These were analyzed using a sliding-window time-varying covariance method. From the autocovariance in each window, the cycle period and height of the first side lobe peak were determined. Rostral VR variability (determined from standard deviation and coefficient of variation of all cycle periods and the mean peak height) was significantly higher than caudal VR variability. FF-FB interruption significantly decreased the rostral VR cycle period and variability but the rostro-caudal gradient remained. The intersegmental delay was also affected. The caudal VR rhythm with NMA in the caudal pool only was slower but more variable than with NMA over the entire cord. These results indicate that the locomotor rhythm in the presence of supraspino-spinal interactions is slower but has a higher variability. The higher variability may reflect a dynamic stability of the system. Additionally, differences in local neural organization likely contribute to rostro-caudal differences in variability of the motor output.

Algorithms↗

PubMed2002

Oral colonization and immune responses to Streptococcus gordonii expressing a pertussis toxin S1 fragment in mice.

The ability of a recombinant Streptococcus gordonii RJM4 expressing the N-terminal 179-amino acid fragment of S1 subunit of pertussis toxin (PT) as a SpaP/S1 fusion protein was tested for oral colonization and immunogenicity in BALB/c mice. Following two consecutive oral inoculations, 100% of the mice were colonized by the recombinant strains for up to 9 weeks and >60% of the animals still retained the strains after 19 weeks. RJM4 recovered from the animals continued to express the SpaP/S1 fusion protein indicating the stability of the fusion gene in vivo. Secretory (S) IgA antibody against PT could be detected in saliva from RJM4-colonized mice but not from the control groups of mice. SIgA against SpaP was also detected in saliva from the RJM4-inoculated and the S. gordonii SL3 (control)-inoculated mice. Serum antibodies against PT and SpaP were not detected in these animals. In conclusion, long-term oral colonization of BALB/c mice with our recombinant S. gordonii was established and the colonization elicited mucosal antibodies against PT and SpaP.

Animals↗

PubMed2002

Regulation of Kv1 subunit expression in oligodendrocyte progenitor cells and their role in G1/S phase progression of the cell cycle.

Proliferative oligodendrocyte progenitor cells (OPs) express large, delayed outward-rectifying K(+) currents (I(K)), whereas nondividing immature and mature oligodendrocytes display much smaller I(K). Here, we show that up-regulation of I(K) occurs in G(1) phase of the cell cycle in purified cultured OPs and is the result of an RNA synthesis-dependent, selective increase of the K(+) channel subunit proteins Kv1.3 and Kv1.5. In oligodendrocyte cells acutely isolated from developing rat brain, a decrease of cyclin D expression is observed as these cells mature along their lineage. This is accompanied by a decrease in Kv1.3 and Kv1.5 subunit expression, suggesting a role for these subunits in the proliferative potential of OPs in situ. I(K) expressed in OPs in subventricular zone and developing white matter in acutely isolated slice preparations were selectively blocked by antagonists of Kv1.3, illustrating the functional presence of this subunit in situ. Interestingly, Kv1.3 block inhibited S-phase entry of both purified OPs in culture and in tissue slice cultures. Thus, we employ both in vitro and in situ experimental approaches to show that (i) RNA-dependent synthesis of Kv1.3 and Kv1.5 subunit proteins occurs in G(1) phase of the OP cell cycle and is responsible for the observed increase in I(K), and (ii) currents through Kv1.3-containing channels play a crucial role in G(1)/S transition of proliferating OPs.

Animals↗

PubMed2002

Cognitive impairment in Parkinson's disease revealed by event-related potential N270.

Disturbed cognitive function is a well-recognized feature of idiopathic Parkinson's disease (PD). The aim of this study was to find a susceptive index to reveal the minor cognitive impairment in PD patients. Thirty PD patients without clinical dementia and thirty-four age-matched normal controls performed a matching task while event-related potentials (ERPs) were recorded from their scalp. There were two kinds of stimulus pairs in this study: match condition, the second stimulus (S2) in a pair was identical to the first one (S1); conflict condition, S2 conflicted with S1 in the color attribute. Subjects were required to press a button in the match condition and another button in the conflict condition. A negative ERP component, N270, which was considered to reflect the conflict processing activity in human brain, was evoked by the S2 of the conflict condition. The patient group showed a delayed and smaller N270 than the control group. The prolongation of its peak latency was significant at P3 and P4 electrodes and the reduction of its mean amplitude was significant at P3 electrode. The amplitude of P300 elicited in the match condition was decreased in the patient group at P4 electrode but its latency did not differ from the control group. These results indicate that PD patients as a group showed cognitive decline even in the absence of clinical dementia. N270 is a sensitive index in revealing this minor cognitive impairment.

Adult↗

PubMed2002

Concurrent measurement of the survival of two populations of rabbit platelets labeled with either two PKH lipophilic dyes or two concentrations of biotin.

BACKGROUND: To avoid radioisotopic labeling and permit comparison of the survival of two platelet populations concurrently in one animal, we compared simultaneous recoveries and survival times of homologous rabbit platelets labeled in vitro with the lipophilic dyes PKH26 (red fluorescing) and PKH67 (green fluorescing) and with two levels of biotin (low, 1 microg/ml; high, 10 microg/ml). METHODS: Blood samples were drawn up to 96 h postinfusion and analyzed by flow cytometry. Biotin-labeled samples were incubated with phycoerythrin-streptavidin before analysis. RESULTS: Recovery of PKH26-labeled platelets at 1 h was lower (37.5%) than that of PKH67-labeled platelets (47.3%; P < 0.001). Platelet survival times were 62.4 and 61.9 h. Recoveries at 1 h of platelets labeled with two levels of biotin were similar (86.6% and 84.6%) and greater than those of PKH-labeled platelets (P < 0.001). Survival of platelets labeled with biotin did not differ (low, 83.3 h; high, 85.2 h) and was longer than for PKH-labeled platelets (P < 0.01). Labeling methods did not activate platelets (measured by P-selectin expression), nor did they affect platelet responses to adenosine diphosphate (ADP), collagen, or thrombin. CONCLUSIONS: Labeling with two levels of biotin is superior to labeling with PKH dyes, and is useful for measuring concurrently the survival of two differing platelet populations.

Adenosine Diphosphate↗

PubMed2002

National study of adverse reactions after vaccination with bacille Calmette-Guérin.

Few large prospective studies of adverse reactions after bacille Calmette-Guérin (BCG) vaccination are available. In a prospective national study of such adverse reactions among 918 subjects (aged 1 day to 54 years) over a 14-month period, 45 vaccinees (5%) reported 53 adverse reactions (23 injection-site abscesses, 14 severe local reactions, 10 cases of lymphadenitis, and 6 other reactions). Only 1% of vaccinees required medical attention. Reactions, particularly lymphadenitis, were significantly less common in infants <6 months old (but not in subjects aged > or =6 months) vaccinated by trained (vs. untrained) providers (relative risk [RR], 0.24; 95% confidence interval [CI], 0.09-0.68). Injection-site abscesses (RR, 2.96; 95% CI, 1.11-7.90) and severe local reactions (RR, 4.93; 95% CI, 1.11-21.90) were significantly more common in older vaccinees. Local reactions were more frequently reported by adult females than by adult males (RR, 7.18; 95% CI, 1.59-32.45). Adverse reactions were not significantly associated with any currently available vaccine batch, previous receipt of BCG vaccine, or concomitant administration of other vaccines.

Adolescent↗

PubMed2002

High mobility group box chromosomal protein 1: a novel proinflammatory mediator in synovitis.

OBJECTIVE: High mobility group box chromosomal protein 1 (HMGB-1) is a ubiquitous chromatin component expressed in nucleated mammalian cells. It has recently and unexpectedly been demonstrated that stimulated live mononuclear phagocytes secrete HMGB-1, which then acts as a potent factor that causes inflammation and protease activation. Macrophages play pivotal roles in the pathogenesis of arthritis. The aim of this study was to determine whether synovial macrophage expression of HMGB-1 is altered in human and experimental synovitis. METHODS: Intraarticular tissue specimens were obtained from healthy Lewis rats, Lewis rats with Mycobacterium tuberculosis-induced adjuvant arthritis, and from patients with rheumatoid arthritis (RA). Specimens were immunohistochemically stained for cellular HMGB-1. Extracellular HMGB-1 levels were assessed in synovial fluid samples from RA patients by Western blotting. RESULTS: Immunostaining of specimens from normal rats showed that HMGB-1 was primarily confined to the nucleus of synoviocytes and chondrocytes, with occasional cytoplasmic staining and no extracellular matrix deposition. In contrast, inflammatory synovial tissue from rats with experimental arthritis as well as from humans with RA showed a distinctly different HMGB-1 staining pattern. Nuclear HMGB-1 expression was accompanied by a cytoplasmic staining in many mononuclear cells, with a macrophage-like appearance and an extracellular matrix deposition. Analysis of synovial fluid samples from RA patients further confirmed the extracellular presence of HMGB-1; 14 of 15 samples had HMGB-1 concentrations of 1.8-10.4 microg/ml. CONCLUSION: The proinflammatory mediator HMGB-1 was abundantly expressed as a nuclear, cytoplasmic, and extracellular component in synovial tissues from RA patients and from rats with experimental arthritis. These findings suggest a pathogenetic role for HMGB-1 in synovitis and indicate a new potential therapeutic target molecule.

Adult↗

PubMed2002

Cellular and molecular alterations in spinal cord injury patients with pressure ulcers: a preliminary report.

The study was designed to investigate the changes, both numerically and functionally, of the molecules critical to wound healing in spinal cord injury (SCI) patients. Spinal cord injury patients who demonstrated delayed healing of their pressure ulcers were used as study subjects. Age-matched healthy individuals served as controls. Adhesion molecule expression of the peripheral blood leukocytes, including lymphocytes and granulocytes, was measured by flow cytometric analysis. Binding capacity of the lymphocytes was evaluated using human umbilical cord vein endothelial cells (HUVECs) as the binding matrix. Samples from pressure ulcers of the patients were immunostained to define fibronectin, kalinin, beta4 integrin, alpha2beta1, alpha3beta1, alpha5beta1, and CD138 expression. Compared to healthy controls, there was decreased expression of CD11a, CD11b, CD18, CD49b, CD49c, CD49d, CD54, and CD8 in patients' lymphocyte populations and CD11a, CD18, CD49c, CD49d, and CD8 in patients' granulocyte populations. The binding capacity, expressed as percentage binding of the lymphocytes to the HUVEC matrix, was greatly diminished in the patients. There was markedly diminished immunohistochemical staining of fibronectin in pressure ulcers. These findings showed that delayed healing of pressure ulcers in SCI patients can be attributed to reduced adhesion molecule expression, impaired cell-cell interaction, and lack of extracellular matrix structural and functional protein.

Adult↗

PubMed2002

Genomic-scale analysis of gene expression profiles in TNF-alpha treated human umbilical vein endothelial cells.

OBJECTIVE AND DESIGN: TNF-alpha is a potent proinflammatory cytokine that plays an important role in immunity and inflammation, and in the control of cell proliferation, differentiation and programmed cell death. However, it is known that TNF-alpha is also the founding member of a still growing family of cytokines with diverse bioregulative functions. Its detailed molecular mechanisms on endothelial activation and injury remain to be elucidated. This study was aimed at determining genomic-scale gene expression profiles in TNF-alpha treated human endothelial cells. MATERIALS AND METHODS: In this study cultured human umbilical vein endothelial cells (HUVECs) were stimulated with TNF-alpha (10 ng/ml) for 2 and 16 h, respectively, and the gene expression pattern was profiled using a cDNA array representing 14,000 gene/cDNA clusters. RESULTS: In total, 72 known human genes were identified the expression levels of which altered over 2-fold in response to TNF-a stimulation. Such alteration was confirmed for IL-8 and MCP-1, with an independent quantitative mRNA assay. It was observed that genes with related biological functions were often temporally co-regulated. CONCLUSIONS: These results indicate the transcriptional pathways mediated by TNF-alpha inside the HUVECs. Expression profiling in HUVECs responding to TNF-alpha stimulation should give an understanding of the molecular mechanisms involved in vascular inflammation.

Cells, Cultured↗

PubMed2002

Mechanism of integration and excision in conjugative transposons.

Translocation of conjugative transposons proceeds via excision of the element to generate a circular molecule that can then integrate into a new site, which can be in the same or a different cell. This review summarises some of the different mechanisms used for excision and integration of conjugative transposons.

Bacteriophages↗

PubMed2002

Mu- and delta-opioid receptor mRNAs are expressed in periaqueductal gray neurons projecting to the rostral ventromedial medulla.

Opioid antinociception appears to be mediated at least in part by a pathway that projects from the periaqueductal gray (PAG) to the rostral ventromedial medulla (RVM), but the relationship between opioid receptors and PAG-RVM projection neurons is unclear. Previous electrophysiological studies have suggested that opioids act directly on some PAG neurons projecting to the RVM. However, immunoreactivity for neither the cloned mu-opioid receptor (MOR1) nor the cloned delta-opioid receptor (DOR1) has been observed in PAG cells retrogradely labeled from the RVM. In the present study, we examined the expression of DOR1 and MOR1 mRNAs in PAG neurons projecting to RVM using quantitative in situ hybridization and retrograde tract-tracing. Mesencephalic neurons were labeled in three male Sprague-Dawley rats by microinjection of Fluoro-Gold into the RVM. Five micrometer cryostat sections were cut and in situ hybridization was performed using full-length cRNA probes labeled with 35S-UTP. Retrogradely labeled neurons that were also labeled for MOR1 or DOR1 mRNA were observed in the dorsomedial, lateral, and ventrolateral portions of the PAG. Quantification was performed in the dorsomedial and ventrolateral PAG using the physical disector. We found that of 219 retrogradely labeled neurons, 50 +/- 14% expressed DOR1 mRNA. In a second set of 120 Fluoro-Gold-labeled neurons, 27 +/- 8% expressed MOR1 mRNA. Significantly more PAG-RVM projection neurons were labeled for MOR1 mRNA in the ventrolateral subregion of the PAG than in the dorsomedial subregion. However, no significant difference was observed in the proportions of retrogradely labeled neurons labeled for DOR1 mRNA in the ventrolateral subregion compared to the dorsomedial subregion. We conclude that opioids are likely to exert direct effects on PAG-RVM projection neurons through both delta- and mu-opioid receptors. In addition, direct effects on PAG-RVM projection neurons from activation of MOR1 appear more likely to be exerted in the ventrolateral PAG than in the dorsomedial PAG.

Animals↗

PubMed2002

Naturally occurring free D-aspartate is a nuclear component of cells in the mammalian hypothalamo-neurohypophyseal system.

It is generally believed that only L-amino acids have a physiological role in species other than bacteria. Recently, the existence of some D-amino acids, particularly D-aspartate, in various organs of several higher animals has been reported. Here we demonstrate that naturally occurring free D-aspartate is localized subcellularly to the heterochromatin in the nucleoli (but not in either the dendrites or axonal terminals) of magnocellular neurosecretory neurons in the rat hypothalamus, and also of microglia and pericytes in the posterior pituitary. Our results imply that naturally occurring free D-aspartate might have a physiological role in nuclear function in mammals. The findings provide new insight for the biological function of D-stereoisomers of amino acids as well as the organization of the nucleus of at least some eukaryotic cells.

Animals↗

PubMed2002

Injection of brain-derived neurotrophic factor in the rostral ventrolateral medulla increases arterial blood pressure in anaesthetized rats.

Brain-derived neurotrophic factor (BDNF) is a unique neurotrophin which not only supports the development of neurons but also modulates the synaptic activity in a number of neuronal systems. BDNF is synthesized in neurons, anterogradely transported and released from nerve terminals and exerts acute effects on synaptic transmission in both peripheral and central nervous systems. Previous studies have shown that BDNF is distributed in several groups of neurons in the brain stem which regulate cardiovascular functions. Here we showed that injection of BDNF (40-400 ng/100 nl) into the rostral ventrolateral medulla resulted in a significant increase in arterial blood pressure (Delta35.5+/-3.5 mmHg) in rats. The duration of change in blood pressure was 145+/-40 s with a latency of 3-5 s. There was no significant effect on the heart rate. The injection of glutamate as a positive control also triggered an increase in blood pressure. Injection of phosphate-buffered saline as a control or the same amount of nerve growth factor did not cause significant changes in blood pressure in different preparations. Immunohistochemistry showed that the nerve terminals immunoreactive for BDNF were localized in several brain stem regions and terminate around spinal projection neurons in the rostral ventrolateral medulla. Neurons in the rostral ventrolateral medulla can uptake exogenous BDNF and express the high affinity receptor trkB. From these results we suggest that BNDF in the medulla may play a role in the regulation of blood pressure.

Amidines↗

PubMed2002

Chronic neuropathic pain is accompanied by global changes in gene expression and shares pathobiology with neurodegenerative diseases.

Neuropathic pain is induced by injury or disease of the nervous system. Studies aimed at understanding the molecular pathophysiology of neuropathic pain have so far focused on a few known molecules and signaling pathways in neurons. However, the pathophysiology of neuropathic pain appears to be very complex and remains poorly understood. A global understanding of the molecular mechanisms involved in neuropathic pain is needed for a better understanding of the pathophysiology and treatment of neuropathic pain. Towards this end, we examined global gene expression changes as well as the pathobiology at the cellular level in a spinal nerve ligation neuropathic pain model using DNA microarray, quantitative real-time PCR and immunohistochemistry. We found that the behavioral hypersensitivity that is manifested in the persistent pain state is accompanied by previously undescribed changes in gene expression. In the DRG, we found regulation of: (1) immediate early genes; (2) genes such as ion channels and signaling molecules that contribute to the excitability of neurons; and (3) genes that are indicative of secondary events such as neuroinflammation. In addition, we studied gene regulation in both injured and uninjured DRG by quantitative PCR, and observed differential gene regulation in these two populations of DRGs. Furthermore, we demonstrated unexpected co-regulation of many genes, especially the activation of neuroinflammation markers in both the PNS and CNS. The results of our study provide a new picture of the molecular mechanisms that underlie the complexity of neuropathic pain and suggest that chronic pain shares common pathobiology with progressive neurodegenerative disease.

Animals↗

PubMed2002

Estrogen receptors alpha and beta in the inner ear of the 'Turner mouse' and an estrogen receptor beta knockout mouse.

Estrogen receptors have earlier been shown in the normal mouse, rat and human inner ear. If estrogens are important in normal hearing and development of presbyacusis in the normal population is not known. However it is known that patients with Turner syndrome, where a lack of estrogens is one of the main characteristics, commonly develop an early presbyacusis. A 'Turner mouse' has been developed, as a model for the ear problems in Turner syndrome, and it shows otitis media and a premature aging of the hearing. Estrogen receptors exist in an alpha and a beta form. In this study inner ear tissue, from the Turner mouse and an estrogen receptor beta knockout mouse (betaERKO), was investigated regarding estrogen receptor alpha and beta using immunohistochemistry. Results show that the Turner mouse has the same pattern of inner ear labeling, both concerning the estrogen receptor alpha and beta, as that of a normal CBA/Ca mouse, with positive staining in the organ of Corti and spiral ganglion. The betaERKO mice show close to normal inner ear morphology and positive estrogen receptor alpha immunostaining at the same locations as the CBA/Ca mouse.

Animals↗

PubMed2002

The chemical syntheses and bioactivities of novel peptide-based endothelin antagonists.

Endothelin antagonists, novel tripeptides containing a series of unnatural amino acids, were synthesized and characterized. A linear peptide BQ-485, perhydroazepin-1-yl-L-leucyl (1)-D-tryptophanyl (2)-D-tryptophan (3), was selected as the parent compound. The introduction of D-Phe derivatives into these peptidic ET antagonists resulted in potent activity against the contraction of rat aortic smooth muscles induced by ET-1 (10 nM) which activated the ET receptors. Among these compounds, 15 tripeptides had high enough antagonistic activity at the level of 10(-7) mol/L (IC50). The activity of three compounds was 10(-6) mol/L (IC50). These HIM-CO-Leu-D-Trp-D-Phe(-R)-OH compounds as ETA antagonists may provide a tool for the development of therapeutic agents in the treatment of putative ET-1-related disorders.

Animals↗

PubMed2002

Melatonin blocks rat hippocampal neuronal apoptosis induced by amyloid beta-peptide 25-35.

To investigate whether melatonin protects neurons from apoptosis. we used amyloid beta-peptide 25-35 (Abeta(25-35)) to induce apoptosis in cultured hippocampal neurons, and monitored the apoptotic activity of the neurons with or without melatonin treatment. Present study shows that melatonin at concentrations of 1 x 10(-6) and 1 x 10(-5) mol/L prevents neuronal morphological changes induced by apoptosis, as characterized by the shrunken and rounded morphology caused by condensed chromatin and fragmented DNA. Melatonin further exhibited the ability to inhibit apoptotic internucleosomal DNA degradation. Moreover, flow cytometric analysis of cell cycle demonstrated that melatonin abolished the Abeta(25-35)-induced apoptotic peak. Our results suggest that melatonin may play an important role to protect neurons from Abeta(25-35)-induced apoptosis.

Amyloid beta-Peptides↗