Search PubMed⌕ Search

Biomedical subjects

D W Wang

Publications and source records attributed to D W Wang.

At least 37 records · Page 2Linked to original sources
PubMed1999

Apoptosis of circulating lymphocytes induced by whole body gamma-irradiation and its mechanism.

We studied the apoptosis of mousecirculating lymphocytes and its mechanism induced by 2, 4, 6, and 8 Gy of whole-body gamma-irradiation and the expression of bax and bcl-2 gene products as related to apoptosis. We found that, in the early stage after irradiation (4th-7th day), the percentage of lymphocyte apoptosis increased rapidly. Four hours after 2, 4, 6, and 8 Gy of gamma-irradiation, the apoptotic lymphocytes were 2.6, 3.8, 5.5, and 10.4 times those in the controls, respectively. A good correlation was found between the intensity of apoptosis and the radiation dose. As the radiation dose increased, the absolute counts of peripheral lymphocytes decreased sharply; 4 hours after 2, 4, 6, and 8 Gy of gamma-irradiation, the lymphocyte counts were 82, 63, 47, and 22% of the controls, respectively. The peak value of lymphocyte apoptosis was observed on the 7th day after irradiation using the in situ terminal labeling method, and the apoptotic lymphocytes were found to be 16 times the number in the controls. These results were in accordance with those obtained by the May-Grünwald-Giemsa staining method. The absolute counts of peripheral lymphocytes dropped to their lowest value on the 7th day after irradiation, suggesting that lymphocyte apoptosis might be the major cause of lymphocytopenia in the early stage after irradiation. The abnormal expression of bax and bcl-2 gene products in irradiated lymphocytes was closely related to apoptosis in peripheral lymphocytes.

Animals↗
PubMed1998

The transient outward current in mice lacking the potassium channel gene Kv1.4.

1. The transient outward current (Ito) plays a prominent role in the repolarization phase of the cardiac action potential. Several K+ channel genes, including Kv1.4, are expressed in the heart, produce rapidly inactivating currents when heterologously expressed, and may be the molecular basis of Ito. 2. We engineered mice homozygous for a targeted disruption of the K+ channel gene Kv1.4 and compared Ito in wild-type (Kv1.4+/+), heterozygous (Kv1.4+/-) and homozygous 'knockout' (Kv1.4-/-) mice. Kv1.4 RNA was truncated in Kv1.4-/- mice and protein expression was absent. 3. Adult myocytes isolated from Kv1.4+/+, Kv1.4+/- and Kv1.4-/- mice had large rapidly inactivating outward currents. The peak current densities at 60 mV (normalized by cellular capacitance, in pA pF-1; means +/- s.e.m.) were 53.8 +/- 5. 3, 45.3 +/- 2.2 and 44.4 +/- 2.8 in cells from Kv1.4+/+, Kv1.4+/- and Kv1.4-/- mice, respectively (P < 0.02 for Kv1.4+/+ vs. Kv1.4-/-). The steady-state values (800 ms after the voltage clamp step) were 30.9 +/- 2.9, 26.9 +/- 3.8 and 23.5 +/- 2.2, respectively (P < 0.02 for Kv1.4+/+ vs. Kv1.4-/-). The inactivating portion of the current was unchanged in the targeted mice. 4. The voltage dependence and time course of inactivation were not changed by targeted disruption of Kv1.4. The mean best-fitting V (membrane potential at 50 % inactivation) values for myocytes from Kv1.4 +/+, Kv1.4+/- and Kv1. 4-/- mice were -53.5 +/- 3.7, -51.1 +/- 2.6 and -54.2 +/- 2.4 mV, respectively. The slope factors (k) were -10.1 +/- 1.4, -8.8 +/- 1.4 and -9.5 +/- 1.2 mV, respectively. The fast time constants for development of inactivation at -30 mV were 27.8 +/- 2.2, 26.2 +/- 5. 1 and 19.6 +/- 2.1 ms in Kv1.4+/+, Kv1.4+/- and Kv1.4-/- myocytes, respectively. At +30 mV, they were 35.5 +/- 2.6, 30.0 +/- 2.1 and 28. 7 +/- 1.6 ms, respectively. The time constants for the rapid phase of recovery from inactivation at -80 mV were 32.5 +/- 8.2, 23.3 +/- 1.8 and 39.0 +/- 3.7 ms, respectively. 5. Nearly the entire inactivating component as well as more than 60 % of the steady-state outward current was eliminated by 1 mM 4-aminopyridine in Kv1.4+/+, Kv1.4+/- and Kv1.4-/- myocytes. 6. Western blot analysis of heart membrane extracts showed no significant upregulation of the Kv4 subfamily of channels in the targeted mice. 7. Thus, Kv1.4 is not the molecular basis of Ito in adult murine ventricular myocytes.

Action Potentials↗
PubMed1998

Improvement of beamline 4B9A and results of an X-ray diffraction experiment.

4B9A is a focusing and monochromatic photon beam at the BSRF, which was constructed in 1990. During the second phase of the BSRF program, the surface of the cylindrical mirror has been coated with Pt, covering the original Ni, and the monochromator has been upgraded. The maximum photon energy extends to 11 keV and the intensity has increased about tenfold with respect to the previous intensity at 6 keV. Synchrotron X-ray diffraction patterns for the Hg-1223 (HgBa(2)Ca(2)Cu(3)O(8+delta)) superconducting bulk and thin film have been measured at 1.54014 A. Results indicate that the bulk and film can be indexed as possessing tetragonal symmetry; lattice parameters a = 3.856 A and c = 15.851 A for the bulk Hg-1223 compound, and a = 3.8517 A and c = 15.8511 A for the film. Their structures are similar.

Journal Article↗
PubMed1998

Febrile seizures and generalized epilepsy associated with a mutation in the Na+-channel beta1 subunit gene SCN1B.

Febrile seizures affect approximately 3% of all children under six years of age and are by far the most common seizure disorder. A small proportion of children with febrile seizures later develop ongoing epilepsy with afebrile seizures. Segregation analysis suggests the majority of cases have complex inheritance but rare families show apparent autosomal dominant inheritance. Two putative loci have been mapped (FEB1 and FEB2), but specific genes have not yet been identified. We recently described a clinical subset, termed generalized epilepsy with febrile seizures plus (GEFS+), in which many family members have seizures with fever that may persist beyond six years of age or be associated with afebrile generalized seizures. We now report linkage, in another large GEFS+ family, to chromosome region 19q13.1 and identification of a mutation in the voltage-gated sodium (Na+)-channel beta1 subunit gene (SCN1B). The mutation changes a conserved cysteine residue disrupting a putative disulfide bridge which normally maintains an extracellular immunoglobulin-like fold. Co-expression of the mutant beta1 subunit with a brain Na+-channel alpha subunit in Xenopus laevis oocytes demonstrates that the mutation interferes with the ability of the subunit to modulate channel-gating kinetics consistent with a loss-of-function allele. This observation develops the theme that idiopathic epilepsies are a family of channelopathies and raises the possibility of involvement of other Na+-channel subunit genes in febrile seizures and generalized epilepsies with complex inheritance patterns.

Amino Acid Sequence↗
PubMed1997

Constitutive and impaired signaling of leptin receptors containing the Gln --> Pro extracellular domain fatty mutation.

Leptin (OB), an adipocyte-secreted circulating hormone, and its receptor (OB-R) are key components of an endocrine loop that regulates mammalian body weight. In this report we have analyzed signal transduction activities of OB-R containing the fatty mutation [OB-R(fa)], a single amino acid substitution at position 269 (Gln --> Pro) in the OB-R extracellular domain that results in the obese phenotype of the fatty rat. We find that this mutant receptor exhibits both ligand-independent transcriptional activation via interleukin 6 and hematopoietin receptor response elements and ligand-independent activation of signal transducer and activator of transcription (STAT) proteins 1 and 3. However, OB-R(fa) is unable to constitutively activate STAT5B and is highly impaired for ligand induced activation of STAT5B compared with OB-R(wt). Introduction of the fatty mutation into a OB-R/G-CSF-R chimera generates a receptor with constitutive character that is similar but distinct from that of OB-R(fa). Constitutive mutant OB-R(fa) receptor signaling is repressed by coexpression of OB-R(wt). The implications of an extracellular domain amino acid substitution generating a cytokine receptor with a partially constitutive phenotype are discussed both in terms of the mechanism of OB-R triggering and the biology of the fatty rat.

Animals↗
PubMed1997

Pharmacological targeting of long QT mutant sodium channels.

The congenital long QT syndrome (LQTS) is an inherited disorder characterized by a delay in cardiac cellular repolarization leading to cardiac arrhythmias and sudden death often in young people. One form of the disease (LQT3) involves mutations in the voltage-gated cardiac sodium channel. The potential for targeted suppression of the LQT defect was explored by heterologous expression of mutant channels in cultured human cells. Kinetic and steady state analysis revealed an enhanced apparent affinity for the predominantly charged, primary amine compound, mexiletine. The affinity of the mutant channels in the inactivated state was similar to the wild type (WT) channels (IC50 approximately 15-20 microM), but the late-opening channels were inhibited at significantly lower concentrations (IC50 = 2-3 microM) causing a preferential suppression of the late openings. The targeting of the defective behavior of the mutant channels has important implications for therapeutic intervention in this disease. The results provide insights for the selective suppression of the mutant phenotype by very low concentrations of drug and indicate that mexiletine equally suppresses the defect in all three known LQT3 mutants.

Anti-Arrhythmia Agents↗
PubMed1997

Expression of transforming growth factor beta in radiation interstitial pneumonitis.

We investigated the changes in transforming growth factor beta 1 (TGF-beta 1) mRNA and TGF-beta 3 protein expression that occur in radiation interstitial pneumonitis. We used TGF-beta 1-cDNA probe in situ hybridization and TGF-beta 3 polyclonal antibody in immunohistochemical techniques. Our results showed that the distribution of TGF-beta 1 mRNA and TGF-beta 3 protein basically coincided in blood vessels, airways, lung parenchyma, and alveolar macrophages. However, bronchial epithelial cells expressed only TGF-beta 3 proteins and no TGF-beta 1 mRNA. We found an increased expression of TGF-beta 1 mRNA and TGF-beta 3 proteins in radiation interstitial pneumonitis.

Animals↗
PubMed1996

Characterization of human cardiac Na+ channel mutations in the congenital long QT syndrome.

The congenital long QT syndrome (LQTS) is an inherited disorder characterized by a prolonged cardiac action potential. This delay in cellular repolarization can lead to potentially fatal arrhythmias. One form of LQTS (LQT3) has been linked to the human cardiac voltage-gated sodium channel gene (SCN5A). Three distinct mutations have been identified in the sodium channel gene. The biophysical and functional characteristics of each of these mutant channels were determined by heterologous expression of a recombinant human heart sodium channel in a mammalian cell line. Each mutation caused a sustained, non-inactivating sodium current amounting to a few percent of the peak inward sodium current, observable during long (> 50 msec) depolarizations. The voltage dependence and rate of inactivation were altered, and the rate of recovery from inactivation was changed compared with wild-type channels. These mutations in diverse regions of the ion channel protein, all produced a common defect in channel gating that can cause the long QT phenotype. The sustained inward current caused by these mutations will prolong the action potential. Furthermore, they may create conditions that promote arrhythmias due to prolonged depolarization and the altered recovery from inactivation. These results provide insights for successful intervention in the disease.

Animals↗
PubMed1996

Comparison of the effects of class I anti-arrhythmic drugs, cibenzoline, mexiletine and flecainide, on the delayed rectifier K+ current of guinea-pig ventricular myocytes.

The effect of class I anti-arrhythmic drugs, cibenzoline, mexiletine and flecainide, on the delayed rectifier potassium current (IK) in guinea-pig ventricular myocytes was studied using whole cell voltage clamp techniques and under blockade of the L-type calcium current by 5 microM nitrendipine. IK consisted of two different current systems, as reported by Sanguinetti and Jurkiewicz (1990), i.e. an E4031-sensitive rapidly activating component (IKr) with a strong inward-going rectification property and an E4031-insensitive slowly activating component (IKs) with little rectification. Cibenzoline (30 microM) decreased both IKr and IKs while flecainide (10 and 30 microM) decreased the IKr exclusively. Mexiletine (30 microM), in contrast, affected neither IKr nor IKs. Since the inhibition of IK(r) and/or IKs prolongs duration of action potentials and refractory periods, class I drugs which also possess the class III effect may have additional effects in treating certain re-entrant arrhythmias.

Action Potentials↗
PubMed1996

Comparison of heterologously expressed human cardiac and skeletal muscle sodium channels.

In this study we have expressed and characterized recombinant cardiac and skeletal muscle sodium channel alpha subunits in tsA-201 cells under identical experimental conditions. Unlike the Xenopus oocyte expression system, in tsA-201 cells (transformed human embryonic kidney) both channels seem to gate rapidly, as in native tissue. In general, hSkM1 gating seemed faster than hH1 both in terms of rate of inactivation and rate of recovery from inactivation as well as time to peak current. The midpoint of the steady-state inactivation curve was approximately 25 mV more negative for hH1 compared with hSkM1. In both isoforms, the steady-state channel availability relationships ("inactivation curves") shifted toward more negative membrane potentials with time. The cardiac isoform showed a minimal shift in the activation curve as a function of time after whole-cell dialysis, whereas hSkM1 showed a continued and marked negative shift in the activation voltage dependence of channel gating. This observation suggests that the mechanism underlying the shift in inactivation voltage dependence may be similar to the one that is causing the shift in the activation voltage dependence in hSkM1 but that this is uncoupled in the cardiac isoform. These results demonstrate the utility and limitations of measuring cardiac and skeletal muscle recombinant Na+ channels in tsA-201 cells. This baseline characterization will be useful for future investigations on channel mutants and pharmacology.

Animals↗
PubMed1996

Distinct local anesthetic affinities in Na+ channel subtypes.

Lidocaine is a widely used local anesthetic and antiarrhythmic drug that is believed to exert its clinically important action by blocking voltage-gated Na+ channels. Studies of Na+ channels from different species and tissues and the complexity of the drug-channel interaction create difficulty in understanding whether there are Na+ channel isoform specific differences in the affinity for lidocaine. Clinical usage suggests that lidocaine selectively targets cardiac Na+ channels because it is effective for the treatment of arrhythmias with few side effects on muscle or neuronal channels except at higher concentrations. One possibility for this selectivity is an intrinsically higher drug-binding affinity of the cardiac isoform. Alternatively, lidocaine may appear cardioselective because of preferential interactions with the inactivated state of the Na+ channel, which is occupied much longer in cardiac cells. Recombinant skeletal muscle (hSkM1) and cardiac sodium channels (hH1) were studied under identical conditions, with a whole-cell voltage clamp used to distinguish the mechanisms of lidocaine block. Tonic block at high concentrations of lidocaine (0.1 mM) was greater in hH1 than in hSkM1. This was also true for use-dependent block, for which 25-microM lidocaine produced an inhibition in hH1 equivalent to 0.1 mM in the skeletal muscle isoform. Pulse protocols optimized to explore inactivated-state block revealed that hSkM1 was five to eight times less sensitive to block by lidocaine than was hH1. The results also indicate that relatively more open-state block occurs in hSkM1. Thus, the cardiac sodium channel is intrinsically more sensitive to inhibition by lidocaine.

Anesthetics, Local↗
PubMed1996

[Experimental study of Salvia miltiorrhiza on prevention of restenosis after angioplasty].

For evaluating the possibility of Salvia miltiorrhiza (SM) on the prevention of arterial restenosis after angioplasty, we investigated the effect of SM on the intimal thickening of air-injured carotid artery of rats. The results showed that the maximal intimal thickness of the injured arteries was much thinner in the treatment group than that in the control group, indicated that SM could prevent experimental restenosis in rat model. Furthermore, we examined the effect of SM on the proliferation of the isolated rabbit arterial smooth muscle cells (SMC) cultivated in vitro. The results showed that SM inhibited the 3H-TdR take-up and proliferation of SMC in a dose-dependent manner. The study raised the possibility that SM could be used to prevent arterial restenosis after angioplasty.

Angioplasty, Balloon, Coronary↗
PubMed1995

Gut-derived endotoxemia and multiple system organ failure following gunshot wounds combined with hemorrhagic shock: an experimental study in the dog.

The purpose of this study was to explore the causative effects of high-velocity bullet injury (5.56 mm, 930 m/sec) combined with hemorrhagic shock on gut-derived endotoxemia and multiple system organ dysfunction or failure. The concentration of endotoxin in both portal and systemic blood was markedly increased after gunshot wounds together with hemorrhagic shock (p < 0.05-0.01). The portal/systemic lipopolysaccharide ratio persistently decreased in the combined wound (CW) group at 24 to 72 hours after injury, and it was much lower in the CW group than that in the single gunshot wound (SW) group. Also, a significant increase of fecal endotoxin content was found in the CW group as compared to the SW group (118.30 +/- 75.36 vs. 8.89 +/- 3.32 micrograms/g, p < 0.01), which was paralleled with the population of gram-negative bacilli in the intestinal tract. The incidence of multiple system organ failure, multiorgan dysfunction (including single organ failure) in the CW group were 30.8% (4 of 13) and 38.5% (5 of 13), respectively. These results suggested that severe trauma could lead to gut origin endotoxemia and bacterial translocation, which might play an important role in the pathogenesis of sepsis and multiple system organ failure following gunshot wounds.

Animals↗
PubMed1995

Abnormalities of K+ and Ca2+ currents in ventricular myocytes from rats with chronic diabetes.

Ionic mechanisms related to the prolongation of cardiac action potential in rats with chronic diabetes mellitus were studied using whole cell voltage-clamp techniques. Diabetes was induced by injection of streptozotocin (STZ; 65 mg/kg body wt) into the tail vein, and ventricular myocytes were isolated from STZ-injected rats (24-30 wk) and from age-matched normal rats. The current densities of transient outward current (Ito), a steady-state outward current, and L-type Ca2+ current (ICa) were significantly smaller in cells from diabetic animals. In addition, the kinetics of Ito of diabetic cells were modified. 1) The decay of Ito was well fitted by a sum of two exponential components in normal cells; there was only one (slow) component in the diabetic cells. 2) The steady-state inactivation curve of Ito in diabetic cells shifted by 5 mV in the negative direction. 3) Recovery from inactivation of Ito was slower in cells from diabetic animals. These alterations in Ito and the steady-state outward current can account for most of the action potential prolongation heretofore documented. The decrease of ICa may possibly be related to the depressed contraction seen in chronic diabetic mellitus.

Animals↗
PubMed1995

Overexpression of c-erbB-2 and p21 oncoproteins in human radiation-induced skin ulcers.

We studied the overexpression of c-erbB-2 and p21 oncoproteins in human radiation-induced skin ulcers using immunohistochemistry. We found that the positive rate of overexpression of c-erbB-2 and p21 oncoproteins was 92.0 and 92.9%, respectively. The overexpression of c-erbB-2 oncoprotein was observed mainly in the cell membrane of squamous epithelial cells and in the cytoplasm of fibroblasts, endothelial cells, leiomyocytes in the media, and in fibrocytes of the adventitia of mesenchymal arterioles. The location of the p21 oncoprotein overexpression was mostly similar to that of c-erbB-2 with stronger staining in the cytoplasm of squamous epithelial cells and weaker staining in mesenchymal arteriolar walls. The overexpression of c-erbB-2 and p21 oncoproteins may be corresponding to the cancer transformation and poor healing of radiation-induced skin ulcers.

Humans↗
PubMed1995

Abnormal location of p16 protein and overexpression of p53 protein in human radiation-induced skin cancer.

To evaluate the role of p16 multiple tumor suppressor (MTS1) and p53 protein in human radiation-induced skin cancer, we examined the expression of p16 and p53 in eight cases using immunohistochemistry. An abnormal location of p16 expression in the cytoplasm was found in seven of eight cases, but the expression of p53 in nuclei was noted in only three of eight cases. These findings suggest that the inactivation of p16 might be related to its binding with another unknown oncoprotein in the cytoplasm and thus lead to carcinogenesis.

Carrier Proteins↗