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D Wang

Publications and source records attributed to D Wang.

At least 433 records · Page 24Linked to original sources
PubMed1999

Purification and characterization of TEF1, a transcription factor that controls the human transforming growth factor-alpha promoter.

Transforming growth factor-alpha (TGF-alpha) is a member of the epidermal growth factor family. It activates signal transduction pathways leading to cell proliferation through the interaction with cell surface epidermal growth factor receptor. The overexpression of TGF-alpha has been found in many types of cancers and is thought to be involved in the genesis and maintenance of these tumors. Recent results also implicate this growth factor in the development of certain diabetic complications, such as atherosclerosis. The function of TGF-alpha can be tightly controlled at the level of transcription of its gene. We have previously characterized the proximal TGF-alpha promoter and identified two neighboring regulatory elements that appeared to cooperate with each other in the regulation of TGF-alpha transcription. The transcription factor that functions through the distal element was identified as AP-2, a protein that was found to be induced by the oncoprotein, Ras. However, what factor binds and controls the proximal regulatory element (PRE) is still unclear. Here, we report the purification and preliminary characterization of the PRE-binding transcription factor TEF1 by sequence-specific DNA-affinity chromatography from rat kidney nuclear extracts. The purified TEF1 migrates on the SDS-PAGE at a molecular mass of about 36 kDa. It specifically interacts with the PRE and was able to strongly activate transcription from the TGF-alpha promoter in HeLa cell nuclear extracts in an in vitro transcription assay. The UV cross-linking experiment confirmed that this 36 kDa protein is indeed the protein that specifically binds the PRE. We also show that the spacing between the AP-2 and the TEF1 sites in the TGF-alpha promoter has little effect on the transcription from the TGF-alpha promoter. The purification of TEF1 furthers our understanding of how TGF-alpha expression is regulated and may help us understand the upstream signaling events that lead to the elevated expression of this growth factor.

Animals↗
PubMed1999

Encapsulation of plasmid DNA in biodegradable poly(D, L-lactic-co-glycolic acid) microspheres as a novel approach for immunogene delivery.

A plasmid DNA encoding bacterial beta-galactosidase gene was encapsulated in poly(d,l-lactic-co-glycolic acid) (PLGA) microspheres. Plasmid DNA extracted from PLGA microspheres retained both structural and functional integrity as evidenced by its restriction endonuclease digestion pattern and its ability to transfect COS-1 cells in vitro. PLGA microspheres protected plasmid DNA from digestion by deoxyribonuclease I (DNase I) in vitro. The encapsulation efficiency of plasmid DNA and its release rate depended on the molecular mass of PLGA. Lastly, J-774A macrophages phagocytosed PLGA microspheres loaded with plasmid DNA. Co-encapsulated monophosphoryl lipid A increased the rate of phagocytosis. These results suggest that biodegradable PLGA microspheres can deliver intact and functional plasmid DNA at controlled rates. Thus, PLGA microspheres may be used to jointly deliver genes and other biologically active molecules, e.g., immunomodulators, to antigen presenting cells.

Animals↗
PubMed1999

Micellar electrokinetic capillary chromatography of psoralen and isopsoralen.

In this paper, a micellar electrokinetic capillary chromatography (MEKC) method to analyze the active components of psoralen and isopsoralen in the Chinese herb Psoralea corylifolia L. was developed. It was found that the optimum separation condition for the analysis of these compounds was a buffer solution which contained phosphate, borate and sodium dodecyl sulfate (pH 9.2). Through optimization of the experimental condition, the two isomers could be baseline-separated and their contents in the methanol-extracted sample were quantitatively analyzed.

Buffers↗
PubMed1999

Effects of nitric oxide synthase inhibitors on systemic hypotension, cytokines and inducible nitric oxide synthase expression and lung injury following endotoxin administration in rats.

Endotoxin shock is characterized by systemic hypotension, hyporeactiveness to vasoconstrictors and acute lung edema. A nitric oxide synthase (NOS) inhibitor, NG-monomethyl-L-arginine (L-NMMA) has been shown to be effective in reversing acute lung injury. In the present study, we evaluated the effects of NOS blockade by different mechanisms on the endotoxin-induced changes. In anesthetized rats, lipopolysaccharide (LPS, Klebsiella pneumoniae) was administered intravenously in a dose of 10 mg/kg. LPS caused sustained systemic hypotension accompanied by an eightfold increase of exhaled NO during an observation period of 4 h. After the experiment, the lung weight was obtained and lung tissues were taken for the determination of mRNA expressions of inducible NOS (iNOS), interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha-(TNF-alpha). Histological examination of the lungs was also performed. In the control group injected with saline solution, mRNA expressions of iNOS, IL-1beta and TNF-alpha were absent. Four hours after LPS, the mRNA expressions of iNOS and IL-1beta were still significantly enhanced, but TNF-alpha was not discernibly expressed. LPS also caused a twofold increase in lung weight. Pathological examination revealed endothelial damage and interstitial edema. Various NOS inhibitors were given 1 h after LPS administration. These agents included Nomega-nitro-L-arginine methyl ester (L-NAME, 10 mg/kg), a constitutive NOS and iNOS inhibitor; S, S'-1,4-phenylene-bis-(1,2-ethanedinyl) bis-isothiourea dihydrobromide (1,4-PBIT, 10 mg/kg), a relatively specific iNOS inhibitor, and dexamethasone (3 mg/kg), an inhibitor of iNOS expression. These NOS inhibitors all effectively reversed the systemic hypotension, reduced the exhaled NO concentration and prevented acute lung injury. The LPS-induced mRNA expressions of iNOS and IL-1beta were also significantly depressed by these NOS inhibitors. Our results suggest that NO production through the iNOS pathway is responsible for endotoxin-induced lung injury. Certain cytokines such as IL-1beta are possibly involved. These changes are minimized by NOS inhibitors through different mechanisms.

Animals↗
PubMed1999

Effects of L-tetrahydropalmatine on energy metabolism, endothelin-1 and NO during acute cerebral ischemia-reperfusion of rats.

To investigate the effects of L-tetrahydropalmatine (L-THP) on energy metabolism, endothelin-1 (ET-1) and NO during acute cerebral ischemia-reperfusion of rats, 24 Wistar rats were randomly divided into four groups, with 6 rats in each group: sham-operation group, simple ischemia group, ischemia-reperfusion group and treatment group (L-THP group). Cerebral ATP, lactate, ET-1 and NO levels were measured in all groups. Our results showed that treatment with L-THP could increase cerebral ATP levels, but decrease cerebral lactate, ET-1 and NO concentrations during ischemia-reperfusion in the treatment group. It is concluded that L-THP could improve cerebral energy metabolism and protect the injured brain tissue, the mechanism of which might be related to suppression of overproduction of ET-1 and NO.

Animals↗
PubMed1999

Effects of temperature and photoperiod on thermogenesis in plateau pikas (Ochotona curzoniae) and root voles (Microtus oeconomus).

We examined the effects of temperature and photoperiod on metabolic thermogenesis and the thermogenic characteristics of brown adipose tissue in plateau pikas (Ochotona curzoniae) and root voles (Microtus oeconomus), the dominant species of small mammals in the alpine meadow ecosystems on the Qinghai-Tibetan Plateau. Pikas and voles were acclimated in the following groups: (1) Long day-warm temperature (16L:8D, 23 degrees C) (2) Long day-cold temperature (16L:8D, 5 degrees C), (3) short day-warm temperature (8L:16D, 23 degrees C), and (4) short day-cold temperature (8L:16D, 5 degrees C). Both temperature and photoperiod were important environmental cues for changes in thermogenesis for both species. Low temperature and short photoperiod induced increases in metabolic rate, nonshivering thermogenesis (NST), mitochondrial protein contents of brown adipose tissue, and cytochrome C oxidase activity of brown adipose tissue mitochondria in both species. Plateau pikas were more sensitive to cold (79% of the total NST response) than to short photoperiod (21%), while root voles were more sensitive to short photoperiod (60% of the total NST response) than to cold (40%), although cold clearly enhanced thermogenesis. Their thermogenic characteristics correlated with their preferred habitats: plateau pikas are found mainly in more exposed microhabitats in open sunny meadow, while root voles live in more sheltered microhabitats in relatively closed shrub. Our results also showed that temperature and photoperiod combined induce thermogenic adjustments in both species in seasonal acclimatization in their alpine meadow macrohabitat.

Acclimatization↗
PubMed1999

Serum prostate-specific antigen and prostate volume predict long-term changes in symptoms and flow rate: results of a four-year, randomized trial comparing finasteride versus placebo. PLESS Study Group.

OBJECTIVES: To determine whether baseline prostate-specific antigen (PSA), in addition to prostate volume, is associated with long-term changes in symptoms and urinary flow rate. METHODS: Three thousand forty men with benign prostatic hyperplasia enrolled in the PLESS trial were randomly assigned to finasteride 5 mg or placebo for 4 years. Symptoms and flow rate were assessed every 4 months, and data were analyzed by dividing the patients into three groups by baseline PSA tertiles (0 to 1.3, 1.4 to 3.2, and 3.3 ng/mL or greater) and baseline prostate volume tertiles (14 to 41, 42 to 57, and 58 to 1 50 mL). RESULTS: After the initial placebo effect, a slow deterioration in symptoms over time was observed in the placebo-treated men with a baseline PSA 1.4 ng/mL or greater. However, placebo-treated men in the lowest PSA tertile (less than 1.4 ng/mL) had sustained symptomatic improvement that was not seen in placebo-treated men in the higher tertiles (P<0.001). In all finasteride-treated groups, there was initial improvement followed by maintenance or continued symptom improvement over time (approximately 3 to 3.5 points by the end of 4 years). The differences in symptom score improvement between placebo and finasteride were marginal for men with baseline PSA levels less than 1.4 ng/mL (P = 0.128) but were highly significant for men with PSA levels 1.4 ng/mL or greater (P<0.001). Urinary flow rate results were similar to those observed for symptoms. Analysis of symptom and flow rate data by prostate volume tertiles in a 10% subset of men yielded similar results, namely a deterioration of symptoms and flow rate in the two higher tertiles treated with placebo (greater than 41 mL) and a sustained improvement in all three groups of finasteride-treated patients. CONCLUSIONS: Baseline PSA and prostate volume are good predictors of long-term symptomatic and flow rate changes. Baseline PSA levels of 1.4 ng/mL or greater and enlarged prostate glands predict the best long-term response to finasteride compared with placebo.

Enzyme Inhibitors↗
PubMed1999

Effect of intravenous calcitonin gene-related peptide antagonist on the laryngeal chemoreflex in piglets.

The laryngeal chemoreflex (LCR) is characterized by mixed apnea and cardiovascular instability and is elicited by applying water to the laryngeal mucosa of developing animals. The LCR may be fatal in very young animals, and the reflex has been postulated as a possible mechanism of sudden infant death syndrome. Several antagonists have been found to alter the severity of the LCR, but the primary neurotransmitters involved in mediating the reflex response are not yet well understood. This study investigates the effect, on the LCR, of the pharmacologic antagonism of calcitonin gene-related peptide (alphaCGRP), a neurochemical found in abundance in the mammalian laryngeal mucosa and its innervating system. The LCR was elicited in 10 mixed breed piglets, 17.7 days of age (12 to 22 days), before and during infusion of alphaCGRP 8-37, a pharmacologic inhibitor of alphaCGRP, and cardiorespiratory and laryngeal responses were compared. The duration of obstructive apnea decreased from 17.9 to 9. 8 seconds (P < 0.03) in the presence of alphaCGRP 8-37. Mean central apnea did not change for the group (P > 0.05), although it was completely inhibited in 2 animals. Cardiovascular changes were not significantly altered by the alphaCGRP inhibitor. alphaCGRP appears to play a regulatory role in the apneic response of the LCR, particularly its obstructive component, but not the cardiovascular response.

Animals↗
PubMed1999

Differences in soft tissue profile changes following mandibular setback in Chinese men and women.

PURPOSE: This study investigated the soft tissue profile changes after surgical correction of mandibular prognathism in Chinese patients and evaluated the sex differences in the ratios of soft tissue to hard tissue change. PATIENTS AND METHODS: Forty-three adult Chinese patients (18 men and 25 women) with mandibular prognathism were treated by intraoral oblique or vertical ramus osteotomy. Lateral cephalometric radiographs were taken presurgically and between 6 to 12 months postsurgically, and specific soft and hard tissue points were digitized and analyzed with a computer-aided system. RESULTS: As a result of surgical setback of the mandible, the soft tissue to hard tissue change ratios were: Li:Ii = 0.71:1, Si:B = 0.90:1, Pg':Pg = 0.94:1 for the males; and Li:Ii = 0.82:1, Si:B = 0.92:1, Pg':Pg = 1.06:1 for the females. CONCLUSION: The changes in soft tissue were closely correlated with the hard tissue movement after surgical setback of mandible. The average ratios of soft tissue to hard tissue change in horizontal direction appear to show a gender difference, which suggests the need for different ratios when predicting the results of orthognathic surgery.

Adult↗
PubMed1999

A 1H MRS study of probable Alzheimer's disease and normal aging: implications for longitudinal monitoring of dementia progression.

In order to evaluate the capability of 1H MRS to monitor longitudinal changes in subjects with probable Alzheimer's disease (AD), the temporal stability of the metabolite measures N-acetylaspartate and N-acetylaspartylglutamate (NA), total Creatine (Cr), myo-Inositol (mI), total Choline (Chol), NA/Cr, mI/Cr, Chol/Cr and NA/mI were investigated in a cohort of normal older adults. Only the metabolite measures NA, mI, Cr, NA/Cr, mI/Cr, and NA/mI were found to be stable after a mean interval of 260 days. Relative and absolute metabolite measures from a cohort of patients with probable AD were subsequently compared with data from a sample of normal older adult control subjects, and correlated with mental status and the degree of atrophy in the localized voxel. Concentrations of NA, NA/Cr, and NA/mI were significantly reduced in the AD group with concomitant significant increases in mI and mI/Cr. There were no differences between the two groups in measures of Cr, Chol, or Chol/Cr. Significant correlations between mental status as measured by the Mini-Mental State Examination and NA/mI, mI/Cr and NA were found. These metabolite measures were also significantly correlated with the extent of atrophy (as measured by CSF and GM composition) in the spectroscopy voxel.

Aged↗
PubMed1999

An evaluation of the time dependence of the anisotropy of the water diffusion tensor in acute human ischemia.

We have performed MRI examinations to determine the water diffusion tensor in the brain of six patients who were admitted to the hospital within 12 h after the onset of cerebral ischemic symptoms. The examinations have been carried out immediately after admission, and thereafter at varying intervals up to 90 days post admission. Maps of the trace of the diffusion tensor, the fractional anisotropy and the lattice index, as well as maps of cerebral blood perfusion parameters, were generated to quantitatively assess the character of the water diffusion tensor in the infarcted area. In patients with significant perfusion deficits and substantial lesion volume changes, four of six cases, our measurements show a monotonic and significant decrease in the diffusion anisotropy within the ischemic lesion as a function of time. We propose that retrospective analysis of this quantity, in combination with brain tissue segmentation and cerebral perfusion maps, may be used in future studies to assess the severity of the ischemic event.

Anisotropy↗
PubMed1999

Reduction of myocardial infarct size by inhibition of inducible nitric oxide synthase.

The inducible nitric oxide synthase isoform (iNOS) is upregulated by cytokines and endotoxins in many types of cells, including cardiac myocytes. Nitric oxide (NO) induced by cytokines can be cytotoxic, and has been implicated in the pathophysiology of myocardial infarction, cardiomyopathy, and septic shock. To examine the role of iNOS in the ischemic myocardium, we studied: 1) the time course of expression of iNOS mRNA after myocardial infarction (MI) in male Sprague-Dawley rat hearts and expression of iNOS protein in the infarcted region; 2) whether hypoxia in vitro is a potential mediator of the induction of iNOS mRNA; and 3) whether inhibition of iNOS by two different selective inhibitors (aminoguanidine and S-methylisothiourea sulfate) in vivo influences infarct size. Myocardial infarction was induced by ligation of the left anterior descending coronary artery (LAD), and tissue was collected at selected times thereafter from both ligated and sham-operated rats. iNOS mRNA was induced in the infarcted region of the left ventricle for 7 days; iNOS protein was also detected in the infarcted area. We next tested whether hypoxia would induce iNOS in vitro. In cultured neonatal ventricular myocytes, iNOS mRNA was slightly induced by 6 to 24 h of hypoxia; however, iNOS protein was only detected when the cytokine interleukin-1beta was present. To study whether iNOS activity contributed to myocardial damage (eg, infarct size), we administered the first dose of the NOS inhibitors 24 h before LAD occlusion and then a second dose after surgery. Inhibition of iNOS activity with aminoguanidine reduced infarct size by 20% but had no effect on infiltration by neutrophils, whereas the more selective inhibitor S-methylisothiourea sulfate reduced infarct size by 41%. These data suggest that NO derived from the iNOS isoform contributes to some of the myocardial injury following MI, possibly by causing myocardial cell death in areas bordering the ischemic region of the heart.

Animals↗
PubMed1999

A novel nociceptor signaling pathway revealed in protein kinase C epsilon mutant mice.

There is great interest in discovering new targets for pain therapy since current methods of analgesia are often only partially successful. Although protein kinase C (PKC) enhances nociceptor function, it is not known which PKC isozymes contribute. Here, we show that epinephrine-induced mechanical and thermal hyperalgesia and acetic acid-associated hyperalgesia are markedly attenuated in PKCepsilon mutant mice, but baseline nociceptive thresholds are normal. Moreover, epinephrine-, carrageenan-, and nerve growth factor- (NGF-) induced hyperalgesia in normal rats, and epinephrine-induced enhancement of tetrodotoxin-resistant Na+ current (TTX-R I(Na)) in cultured rat dorsal root ganglion (DRG) neurons, are inhibited by a PKCepsilon-selective inhibitor peptide. Our findings indicate that PKCepsilon regulates nociceptor function and suggest that PKCepsilon inhibitors could prove useful in the treatment of pain.

Acetic Acid↗
PubMed1999

Effect of lidocaine on improving cerebral protection provided by retrograde cerebral perfusion: a neuropathologic study.

OBJECTIVE: To determine whether lidocaine can improve the neuropathologic results in canine brains after retrograde cerebral perfusion (RCP). DESIGN: Randomized, blinded, experimental study. SETTING: University animal laboratory. PARTICIPANTS: Mongrel dogs. INTERVENTIONS: Fourteen mongrel dogs were placed on 120 minutes of hypothermic (20 degrees C) RCP. Following the RPC, they then resumed cardiopulmonary bypass and rewarming for 60 minutes. In the lidocaine group (n = 8), lidocaine was administered continuously; in the control group (n = 6), normal saline was administered. Cerebral perfusion fixation was performed at the end of the experiment. MEASUREMENTS AND MAIN RESULTS: The number of ischemic cells in 200 neurons was counted in the parietal cortex, CA1 sector of the hippocampus, CA3 sector of the hippocampus, ventral posterolateral nucleus of the thalamus, and Purkinje cells of the cerebellar cortex. Those in the parietal cortex, CA1 sector of the hippocampus, and ventral posterolateral nucleus of the thalamus were significantly less in the lidocaine group than in the control group (25.8+/-17.3 v 53.7+/-12.0; p < 0.01; 17.0+/-8.5 v 54.7+/-22.1; p < 0.01; and 16.9+/-17.8 v 49.7+/-28.4; p < 0.05, respectively). The total number of ischemic cells in the five examined regions was also significantly less in the lidocaine group than in the control group (89.5+/-19.4 v 219.5+/-45.5; p < 0.01). CONCLUSION: Continuous lidocaine significantly alleviated the ischemic neuropathologic injury after RCP and thus possibly improved cerebral protection.

Anesthetics, Local↗
PubMed1999

Stat5 is required for IL-2-induced cell cycle progression of peripheral T cells.

Many cytokines activate two highly homologous Stat proteins, 5a and 5b. Mice deficient in both genes lack all growth hormone and prolactin functions but retain functions associated with cytokines such as erythropoietin. Here, we demonstrate that, while lymphoid development is normal, Stat5a/b mutant peripheral T cells are profoundly deficient in proliferation and fail to undergo cell cycle progression or to express genes controlling cell cycle progression. In addition, the mice lack NK cells, develop splenomegaly, and have T cells with an activated phenotype, phenotypes seen in IL-2 receptor beta chain-deficient mice. These phenotypes are not seen in mice lacking Stat5a or Stat5b alone. The results demonstrate that the Stat5 proteins, redundantly, are essential mediators of IL-2 signaling in T cells.

Animals↗
PubMed1999

Synthesis of a spin-labeled photoaffinity ATP analogue, and its use to specifically photolabel myosin cross-bridges in skeletal muscle fibers.

A spin-labeled photoaffinity ATP analogue 3'(2')-O-[4-[4-oxo-(4-amido-2,2,6,6-tetramethyl-piperidino-1-oxyl)]-benz oyl]benzoyl adenosine 5'-triphosphate (SL-Bz2ATP) was synthesized and used to photolabel myosin in muscle fibers. Previous work has shown that 3'(2')-O-(4-benzoyl)benzoyl adenosine 5'-triphosphate (Bz2ATP) photolabeled Ser-324 of the 50 kDa tryptic fragment of skeletal S1 heavy chain. In this work, [alpha-32P]SL-Bz2ATP was hydrolyzed and trapped as the diphosphate analogue with Co2+ and orthovanadate at the active site of myosin in rabbit psoas muscle fibers. After UV irradiation, the myosin heavy chain was the only protein band found to be significantly photolabeled as assayed by gel electrophoresis and radioactivity counting. The labeling was localized after brief trypsin digestion by SDS-PAGE to be on the 50 kDa tryptic fragment of the S1 heavy chain. Ca. 35% of the myosin in fibers was covalently photolabeled. The fibers photolabeled with SL-Bz2ATP had the same active tension and maximum shortening velocity as the control fibers. The resulting spin label on myosin was too mobile to report the orientation of the heads in fibers. Nonetheless, this is the first work to show the feasibility of utilizing active site binding and photoaffinity labeling to place covalent spectroscopic probes at the myosin active site in fibers with high specificity and yield without affecting mechanical function.

Adenosine Triphosphate↗
PubMed1999

Defective glucose transport across brain tissue barriers: a newly recognized neurological syndrome.

Impaired glucose transport across brain tissue barriers causes infantile seizures, developmental delay and acquired microcephaly. Since the first report in 1991 (De Vivo et al, NEJM, 1991) 17 patients have been identified with the glucose transporter protein syndrome (GTPS). The diagnostic feature of the syndrome is an unexplained hypoglycorrhachia in the clinical setting of an infantile epileptic encephalopathy. We review our clinical experience by highlighting one illustrative case: a 6-year old girl who presented at age 2 months with infantile seizures and hypoglycorrhachia. The CSF/blood glucose ratio was 0.33. DNA sequencing identified a missense mutation in exon 7 (C1108T). Erythrocyte GLUT1 immunoreactivity was normal. The time course of 3-O-methyl-glucose (3OMG) uptake by erythrocytes of the patient was 46% that of mother and father. The apparent Km was similar in all cases (2-4 mmol/L), but the apparent Vmax in the patient was only 28% that of the parents (500 versus 1,766 fmol/s/10(6)RBC; p < 0.004). In addition, a 3-month trial of oral thioctic acid also benefited the patient and increased the Vmax to 935 fmol/s/10(6) RBC (p < 3 x 10(-7)). Uptake of dehydroascorbic acid by erythrocytes of the patient was impaired to the same degree as that of 3OMG (Vmax was 38% of that of the mother's), which supports previous observations of GLUT1 being multifunctional. These studies confirm the molecular basis of the GTPS and the multifunctional role of GLUT1. The need for more effective treatment is compelling.

3-O-Methylglucose↗