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Biomedical subjects

D P Wang

Publications and source records attributed to D P Wang.

At least 19 recordsLinked to original sources
PubMed2006

Hypoxia-inducible factor 1alpha cDNA cloning and its mRNA and protein tissue specific expression in domestic yak (Bos grunniens) from Qinghai-Tibetan plateau.

Adaptation to hypoxia is regulated by hypoxia-inducible factor 1 (HIF-1), a heterodimeric transcription factor consisting of an oxygen-regulated alpha-subunit and a constitutively expressed beta-subunit. How animals living on Qinghai-Tibetan plateau adapt to the extreme hypoxia environment is known indistinctly. In this study, the Qinghai yak, which has been living at 3000-5000 m altitude for at least two millions of years, was selected as the model of high hypoxia-tolerant adaptation species. The HIF-1alpha ORFs (open reading frames) encoding for two isoforms of HIF-1alpha have been cloned from the brain of the domestic yak. Its expression of HIF-1alpha was analyzed at both mRNA and protein levels in various tissues. Both its HIF-1alpha mRNA and protein are tissue specific expression. Its HIF-1alpha protein's high expression in the brain, lung, and kidney showed us that HIF-1alpha protein may play an important role in the adaptation to hypoxia environment.

Amino Acid Sequence↗
PubMed2006

Leptin cDNA cloning and its mRNA expression in plateau pikas (Ochotona curzoniae) from different altitudes on Qinghai-Tibet Plateau.

Leptin, an adipocyte-derived hormone, plays an important role in body energy homeostasis. Plateau pika (Ochotona curzoniae), an endemic and keystone species living only at 3000-5000 m above sea level on Qinghai-Tibet Plateau, is a typically high hypoxia and low temperature tolerant mammal with high resting metabolic rate (RMR), non-shivering thermogenesis (NST), and high ratio of oxygen utilization to cope with harsh plateau environment. To explore the molecular mechanism of ecological acclimation in plateau pika, we first cloned pika leptin cDNA and compared its mRNA expression in different altitudes (3200 and 3900 m) using real-time RT-PCR (Taqman probe) technology. The full-length pika leptin cDNA was 3015 with 504 bp open-reading frame encoding the precursor peptide of 167 amino acids including 21 residues of signal peptide. Pika leptin was 70-72% homologous to that of other species and was of similarly structural characteristics with other species. The pika-specific genetic diversity in leptin sequence occurred at twenty sites. With the increase in altitude, there were larger fat store and high level of ob gene expression in plateau pika. Our results indicated that leptin is sensitive to cold and hypoxia plateau environment and may play one of important roles in pika's ecological adaptation to harsh plateau environment.

Altitude↗
PubMed2005

Induction of systemic lupus erythematosus-like syndrome in syngeneic mice by immunization with activated lymphocyte-derived DNA.

OBJECTIVES: Systemic lupus erythematosus (SLE) is the prototype of autoimmune disease and the mechanisms underlying the disease have not yet been elucidated. Thus, animal models of SLE would facilitate investigation of pathogenetic mechanisms involved in the development of the disease. This study characterizes a murine model of SLE-like syndrome induced by syngeneic activated lymphocyte-derived DNA (referred to as ALD DNA). METHODS: Normal BALB/c mice were immunized subcutaneously with highly purified ALD DNA. Anti-double-stranded DNA (anti-dsDNA) antibodies were determined by enzyme-linked immunosorbent assay. Other SLE-associated autoantibodies were examined by indirect immunofluorescence and anti-ENA (extractable nuclear antigen) profile assay. Pathological changes were analysed by light microscopy and electron microscopy. Kidney cryostat sections were viewed by immunofluorescence for the presence of glomerular IgG and C3 deposits. Proteinuria was measured by Coomassie brilliant blue assay. RESULTS: High levels of anti-dsDNA antibodies and other autoantibodies frequently appearing in SLE were detectable in the sera of ALD DNA-immunized mice. Glomerulonephritis and glomerular deposition of IgG plus C3 were observed in the kidney sections. Moreover, proteinuria was seen in the immunized mice. CONCLUSIONS: SLE-like syndrome can be induced by ALD DNA in normal mice. This induced model may be useful for elucidating the mechanisms involved in autoimmunity to DNA and the development of SLE.

Animals↗
PubMed2001

Stability of 4-DMAP in solution.

A stability-indicating reversed-phase performance liquid chromatographic method was developed for the detection of 4-(N,N-dimethylamino)phenol (4-DMAP) and its degradation products under accelerated degradation conditions. The degradation kinetics of 4-DMAP in aqueous solution over a pH range of 1.12-6.05 and its stability in solutions based on propylene glycol or polyethylene glycol 400 were investigated. The observed rate constants were shown to follow, apparent first-order kinetics in all cases. The pH rate profile shows that maximum stability of 4-DMAP was observed in the pH range 2.0 to 3.0. Acid/base catalysis of 4-DMAP was not affected by systems of various ionic strengths. Incorporation of nonaqueous propylene glycol or polyethylene glycol 400 in the pH 3.05 solution of 4-DMAP showed an increase in the stability at 55 degrees C +/- 0.5 degrees C.

4-Aminopyridine↗
PubMed2001

[Study on in vitro culture and cryopreservation by vitrification of blastocysts derived from single mouse 2-cell embryos' blastomeres].

The present experiments were designed to study the effects of glucose, EDTA, glutamine on the in vitro development of single blastomeres from 2-cell embryos in mouse, and the efficiency of cryopreservation of blastocysts from single blastomers with different vitrification. Single blastomeres derived from female ICR x male BDF1 2-cell embryos were cultured in mKRB with or without glucose, EDTA and glutamine, respectively. The expanded blastocyst rates were significantly different between in mKRB with glucose and without glucose (34% vs 65%); The blastomeres were cultured in mKRB with EDTA and glutamine but glucose, the expanded blastocyst rate (90%) was significantly higher than other groups. The blastocysts derived from single blastomeres were vitrified in liquid nitrogen after equilibration in GFS40 for 0.5-2 min, the survival rate 24%-51%. The blastocysts were pretreated in mPBS with 10% glycerol for 5 min, followed by exposure to GFS40 at 25 degrees C for 0.5 min, then vitrified in liquid nitrogen(two-step method), the survival rate was 61%. However, the survival rates increased to 64% and 70% when the blastocysts were vitrified(one-step method) ater equilibration in EFS40 at 25 degrees C for 0.5-1 min.

Animals↗
PubMed2000

Formulation optimization of controlled-release pellets of metoclopramide hydrochloride using dissolution fit factor approach.

The purpose of this study was to optimize the formulation variables for the preparation of ethyl cellulose-coated nonpareils loaded with metoclopramide hydrochloride (MCL). The approach to evaluate the effectiveness of formulation parameters was monitored by release rate testing using dissolution fit factors as a tool. The content of ethyl cellulose used in the formulation was based on the drug-loaded weight. The interrelationship of each developed formulation and the reference formulation Gastro-Timelets and their respective dissolution curves were evaluated using Moore's equation: [equation: see text]. The relationship between the ethyl cellulose content in the formulation and the dissolution fit factor f2 can be described as the following regression equation: Y = -0.054X2 + 3.347X - 1.915 (r2 = 0.99). The optimum ethyl cellulose content obtained from the equation was 30.8%. The type and content of plasticizer used in the formulation to achieve the greatest f2 were determined to be Myvacet 9-40 at the concentration of 25%. Results indicated that using the release rate testing approach with the dissolution fit factor as a tool could provide valuable information for formulation optimization.

Cellulose↗
PubMed2000

[Effects of hypotonic stress on the survival of hatched mouse blastocysts pericryopreservation].

The objective of this study to evaluate the effect of hypotonic stress on developmental potential of hatched blastocysts perivitrification. Hatched mouse blastocysts were vitrified in liquid nitrogen after equilibration in 10% or 20% GL for 5 min and in GFS40 for 30 sec respectively, the survival rates were 93%-97% after the frozen-thawed embryos were cultured in vitro for 24 h. There were no statistical difference between the frozen and the fresh group (P > 0.05). In order to evaluate effects of hypotonic stress on developmental abilities, fresh hatched mouse blastocysts were respectively exposed to 1.00 x, 0.50 x, 0.30 x, 0.25 x and 0.20 x PBS for 30 min, then cultured in mKRB for 24 h, the survival rates were 98%, 99%, 92%, 92% and 50% respectively. The rate in 0.20 X PBS group was significantly lower than in other groups (P < 0.01). When frozen-thawed embryos were directly treated with different osmotic solutions, the survival rates were 88%, 72%, 58%, 11% and 0 respectively in 1.00 x, 0.50 x, 0.30 x, 0.25 x and 0.20 x PBS group. The rate in 1.00 x PBS group was significantly higher than in other groups (P < 0.05). However, when frozen-thawed embryos were first cultured in vitro for 12 h, then exposed to 1.00 x, 0.50 x, 0.30 x, 0.25 x and 0.2 x PBS, the survival rates were 98%, 94%, 82%, 58% and 26% respectively. There was no statistical difference between 1.00 x and 0.50 x PBS group (P > 0.05). Although the rate in 0.30 x, 0.25 x and 0.20 x PBS group was significantly lower than in 1.00 x group(P < 0.01), it was significantly higher than in the same treatment group without in vitro culture(P < 0.05).

Animals↗
PubMed1999

Formulation development of allopurinol suppositories and injectables.

Allopurinol was formulated into injectable and suppository dosage forms. The injectable formulation was prepared by dissolving allopurinol in a cosolvent system consisting of dimethyl sulfoxide (DMSO) and propylene glycol (v/v = 50/50). The stability of allopurinol in the cosolvent system was studied under accelerated storage conditions, and results indicate first-order degradation kinetics with an activation energy of 24.3 kcal/mol. The development of suppository dosage forms was performed by formulating allopurinol with polyethylene glycol (PEG) mixtures of different molecular weights. In vitro release profiles of suppositories formulated with different polyethylene bases were obtained in the pH 7.4 buffer solution using the USP 23 paddle method at 100 rpm. Results indicate that the release rate of the suppository formulations containing PEG 1500/PEG 4000 at the ratio (w/w) of 2.5/10 to 10/2.5 appeared to be similar. However, the addition of sodium lauryl sulfate in the suppository decreased the release rate of allopurinol significantly. A future study to establish in vitro/in vivo correlation (iv/ivc) is suggested.

Allopurinol↗
PubMed1999

Percutaneous transport of diclofenac sodium from mixtures of fatty alcohol (or fatty acid) and propylene glycol through the rabbit abdominal skin.

Diclofenac sodium is a nonsteroidal anti-inflammatory drug with analgesic, antipyretic, and anti-inflammatory activity. When used in a topical application, diclofenac can diffuse through the skin and into the subcutaneous tissue to effect the anti-inflammatory action. In this study, in vitro evaluations of the percutaneous transport of diclofenac sodium in various bases containing fatty alcohols/propylene glycol or fatty acid/propylene glycol mixtures through the abdominal skin of the rabbit were investigated. Results show that the transdermal flux of diclofenac sodium in the fatty alcohol/propylene glycol bases of the same ratio is affected by the chain length of the fatty alcohol, and its effect is in the order of C10 > C12 > C14 > C18. However, the transdermal flux of diclofenac sodium in the fatty acid/propylene glycol bases of the same ratio is also affected by the chain length of the fatty acid, but no absolute relationship was found. For the same chain length of fatty acid and fatty alcohol used in the formulation base that was otherwise the same, the transdermal flux of diclofenac sodium is higher in the formula containing fatty alcohol than that containing fatty acid.

Abdomen↗
PubMed1999

Uncv (uncovered): a new mutation causing hairloss on mouse chromosome 11.

A pair of mutant mice with a first sparse coat appeared spontaneously in the production stock of BALB/c mice with a normal coat. After being sib-mated, they produced three phenotypes in their progeny: mice with normal hair, mice with a first sparse coat and then a fuzzy coat, and uncovered mice. Genetic studies revealed the mutants had inherited an autosomal monogene that was semi-dominant. By using 11 biochemical loci--Idh, Car2, Mup1, Pgm1, Hbb, Es1, Es10, Gdc, Ce2, Mod1 and Es3--as genetic markers, two-point linkage tests were made. The results showed the gene was assigned to chromosome 11. The result of a three-point test with Es3 and D11Mit8 (microsatellite DNA) as markers showed that the mutation was linked to Es3 with the recombination fraction 7.89 +/- 2.19%, and linked to D11Mit8 with the recombination fraction 26.30 +/- 3.57%. The recombination fraction between Es3 and D11Mit8 was 32.90 +/- 3.81%. It is suggested that the mutation is a new genetic locus that affected the skin and hair structure of the mouse. The mutation was named uncovered, with the symbol Uncv. Further studies showed the mutation affected not only the histology of skin and hair but also the growth and reproductive performance of the mice. The molecular characterization of the Uncv locus needs to be further studied.

Alopecia↗
PubMed1998

Therapeutic effect and pharmacokinetics of ketotifen transdermal delivery system.

Ketotifen transdermal delivery systems were prepared using polyisobutylene, liquid paraffin, and fatty acid. In vitro skin penetration studies were conducted in Franz diffusion cells using excised porcine skin to determine the skin permeation rates of ketotifen patches. A trend of increased skin penetration of ketotifen was observed as the amount of liquid paraffin in the patch was increased. In addition, we found that lauric acid was a suitable enhancer for percutaneous absorption of ketotifen. Challenge tests were performed in guinea pigs to determine the therapeutic effect of the delivery systems for the inhibition of anaphylactic shock using varied concentrations of chicken ovum albumin as sensitizer. Our results showed that compared with the treatment of intramuscular administration, the skin patch was more effective and produced higher survival rates. The pharmacokinetics of the ketotifen patch were determined by applying the skin patch to the dorsal skin of rabbits. The plasma levels were maintained constant (42.5-36.4 ng/ml) from 9 to 30 hr. From our study, the prepared ketotifen patch may further be developed for the treatment or prevention of allergic asthma.

Adhesives↗
PubMed1998

[Localization of the gene for 4 hereditary multiple exostoses families].

We investigated 11 families with hereditary multiple exostoses (EXT) by linkage analysis using 8 short-tandem-repeat (CA)n polymorphic markers on chromosomes 8, 11 and 19. The Lod score in four families indicated that the gene responsible for EXT is located in the pericentromeric region of chromosome 11.

Chromosome Mapping↗
PubMed1996

Transcriptional regulation of the human cholesterol 7 alpha-hydroxylase gene (CYP7A) in HepG2 cells.

A stable HepG2 cell line harboring a human cholesterol 7 alpha-hydroxylase (CYP7A) minigene/luciferase reporter gene construct was selected for studying transcriptional regulation of CYP7A gene promoter. Insulin and phorbol 12-myristate-13-acetate (PMA) strongly repressed the promoter activity as measured with luciferase activity expressed in the cells. The promoter activity of the 5' progressive deletion/luciferase reporter gene constructs was studied in a transient transfection assay in HepG2 cells. PMA represses the promoter activity and the response elements were localized in the -184/-151 and -134/-81 regions. Insulin also represses the promoter activity and response element was mapped in the -298/-81 region. Surprisingly, glucocorticoid receptor (GR) strongly inhibited promoter activity in the presence of dexamethasone, and response elements were localized in the -298/-151 and the -150/+24 regions. Thyroid hormone receptor also repressed promoter activity and response elements were localized in the -150/+24 and upstream regions. Cotransfection of CYP7A chimeric constructs with an expression vector carrying liver-enriched transcription factor HNF3 alpha stimulated the reporter gene activity, but cotransfection with GR plasmid interfered with the HNF3 alpha-stimulated activity possibly through competition for binding to overlapping GR/HNF3 binding sites. Thus, human cholesterol 7 alpha-hydroxylase gene promoter is strongly repressed by insulin, PMA, and steroid/thyroid hormones and results in the low level of cholesterol 7 alpha-hydroxylase expression in the human liver.

Base Sequence↗