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Biomedical subjects

H W Wang

Publications and source records attributed to H W Wang.

At least 19 recordsLinked to original sources
PubMed2006

Relationships among acylation stimulating protein, adiponectin and complement C3 in lean vs obese type 2 diabetes.

OBJECTIVE: The purpose of this study was to determine the relationships between adipocyte hormones acylation stimulating protein (ASP), adiponectin, complement C3 (C3) (ASP precursor) and insulin, C-reactive protein (CRP), lipid profiles and insulin resistance in lean vs obese type 2 diabetes subjects. SUBJECTS: Lean type 2 diabetes subjects (DL n = 27) vs obese type 2 diabetes subjects (DO n = 55) were compared to age-matched nondiabetic groups (Obese, OB n = 55 and control, CTL n = 50). RESULTS: The DO group demonstrated significant increases in plasma ASP and C3 with decreases in plasma adiponectin as compared to CTL. Interestingly, these increases in ASP and C3 were as high, or greater, in the DL group in spite of normal weight. By contrast adiponectin in the DL group was comparable to CTL, in spite of marked insulin resistance. C3 correlated with insulin, glucose and homeostasis model assessment of insulin resistance (HOMA-IR); ASP correlated with body mass index (BMI), glucose, insulin and plasma lipid parameters (non-esterified fatty acids (NEFA), triglyceride, cholesterol and apolipoprotein B). Adiponectin correlated with BMI, glucose, NEFA, triglyceride, high-density lipoprotein cholesterol and apolipoprotein A1 but not HOMA-IR, ASP or C3. CRP correlated only with HOMA-IR. CONCLUSION: Increased ASP and C3 are both associated with diabetes and related lipid factors but are not regulated coordinately. Adiponectin appears to be more closely related to body size (decreased in obese subjects) than insulin resistance in these subjects.

Adiponectin↗
PubMed2005

Engineering and functional evaluation of a single-chain antibody against HIV-1 external glycoprotein gp120.

The HIV-1 envelope glycoprotein surface subunit gp120 is an attractive target for molecular intervention. This is because anti-HIV-1 gp120 neutralizing antibodies display the potential ability to inhibit HIV-1 infection. The present investigation describes the construction of a genetically engineered single chain antibody (scFv102) against HIV-1 gp120, its expression and functional evaluation. The parental hybridoma cell line (102) produces an immunoglobulin directed against the conserved CD4-binding region of gp120. cDNAs encoding the variable regions of the heavy (V(H)) and light (V(L)) chains were prepared by reverse transcription PCR and linked together with an oligonucleotide encoding a linker peptide (Gly(4)Ser)(3) to produce a single chain antibody gene. The resulting DNA construct was cloned into a prokaryotic expression vector (pET28) and recombinant scFv102 was expressed in Eserichia coli as an insoluble protein. The denatured scFv102 was refolded and purified by immobilized metal ion affinity chromatography. Purified scFv102 had the same specificity as the intact IgG in immuno-blotting assays and immuno-fluorescence (IF) detection, but ELISA analyses demonstrated the affinity of scFv102 to be 5-fold lower than that of the parental monoclonal antibody. In neutralization assays, scFv102 at concentrations lower than 40 microg/ml exhibited efficient interference with viral replication and inhibition of viral infection (90%) across a range of primary isolates of subtype B HIV-1. These results suggest that the constructed anti-HIV-1 gp120 scFv102 has good biological activity and can potentially be used for in vitro diagnostic and in vivo therapeutic applications.

Amino Acid Sequence↗
PubMed2004

Topical 5-aminolaevulinic acid-photodynamic therapy for the treatment of urethral condylomata acuminata.

BACKGROUND: Electrocoagulation and laser evaporation for urethral condylomata acuminata have high recurrence rates and can be associated with urethral malformations. OBJECTIVES: To investigate the effect of photodynamic therapy (PDT) with topical 5-aminolaevulinic acid (ALA) on urethral condylomata acuminata and to examine the histological changes in lesions of condylomata acuminata after ALA-PDT. METHODS: Patients with urethral condylomata (n = 164) were given topical ALA followed by intraurethral PDT through a cylindrical fibre. Patients included 11 individuals with 16 penile or vulval condylomatous lesions which were biopsied before or after treatment; the histological changes were then evaluated by light microscopy and electron microscopy. RESULTS: The complete response rate was 95% and the recurrence rate was 5% after 6-24 months of follow-up. Light microscopy revealed keratinocytes in the middle and upper layers of the epidermis showing marked vacuolation and some necrocytosis 1 and 3 h after PDT. Necrosis in all layers of the epidermis was noted 5 h after PDT. Electron microscopy of keratinocytes revealed distinct ultrastructural abnormalities of mitochondria and the endoplasmic reticulum, and membrane damage. Apoptotic bodies were detected 3 h after PDT and a large number of keratinocytes exhibited necrosis 5 h after PDT. CONCLUSIONS: Results suggest that, compared with conventional therapies, topical ALA-PDT is a simple, effective, safe and well-tolerated treatment for urethral condylomata acuminata that is associated with a low recurrence rate. The mechanism might be the triggering of both apoptosis and necrosis by ALA-PDT in human papillomavirus-infected keratinocytes.

Adult↗
PubMed2002

Correlation between voice handicap index and voice laboratory measurements in dysphonic patients.

Factors underlying voice disorders can be categorized into three distinct domains: emotional, physical, and functional. The Voice Handicap Index (VHI) subjectively evaluates voice disorders in terms of these three factors. On the other hand, Voice Laboratory Measurements (VLM) use objective criteria to evaluate the severity of voice disorders. Use of these two different tests (VHI and VLM) on dysphonic patients has, however, tended to yield results that vary widely in their conclusions. This report reviewed 135 testing sessions on dysphonia patients. Seventy-nine of the tests were VHI, and 56 were VLM. All VHI and VLM parameters were entered into a statistical program and analyzed using a Pearson correlation. The results show that each VHI parameter provides a significant level of reliability (P < 0.01) when compared with other VHI parameters. Four VLM parameters also demonstrated significant reliability (P < 0.01) in comparison with other VLM parameters. However, when comparing across testing methods, VHI and VLM parameter reliability is shown to be poor (P > 0.05). With such a large discrepancy between the results of VHI and VLM testing, no objective parameter can yet be regarded as a definitive prognostic factor in a subjective evaluation of dysphonic patients.

Adolescent↗
PubMed2001

Dissociation and subunit rearrangement of membrane-bound human C-reactive proteins.

As one of the most important acute-phase reactants in human serum, C-reactive protein plays its physiological roles mainly on membranes. Here we show that the human C-reactive protein is two-dimensionally crystallized upon specific adsorption on the phosphorylcholine ligand containing membranes by monolayer approach. The 2.0-nm resolution projection structure of the two-dimensional crystals analyzed by electron microscopy and image reconstruction reveals open-ring-like pentamers in the crystals. The electron microscope graphs also show that the dissociated pentamers with open-ring-like structure occur in a closed packing region (not two-dimensionally crystallized). These results indicate a membrane-induced dissociation and rearrangement of hCRP, which may relate to the variety of hCRP's physiological functions.

C-Reactive Protein↗
PubMed2001

Two-dimensional assembly of pentameric rabbit C-reactive proteins on lipid monolayers.

The problem of pentamer packing on a two-dimensional plane is of concern not only in physics and mathematics but also in biology. The packing styles of pentamers may either be related to or reflect the physiological or biochemical properties of biological macromolecules. C-reactive protein (CRP), one of the classical members of the petraxin family, was recently two-dimensionally (2D) crystallized by us on lipid monolayers by specific adsorption (Wang, H. W., and Sui, S. F., 1999, J. Struct. Biol. 127, 283-286). Another type of the protein's 2D crystal under the same conditions was obtained in the present work. The new 2D crystal was studied using electron microscopy of negatively stained specimens followed by image processing. A projection map at 2.2-nm resolution was obtained. The previous 2D crystal (PI) and the current 2D crystal (PII) show different pentamer-packing styles. Both of them are closely related to the fivefold symmetry of the molecule itself. The coexistence and the spatial contiguity of the two types of pentamer assembly were observed in a visual field. The fivefold symmetrical macromolecule can form a pentiling pattern on a two-dimensional plane, which has never been reported in biological system before. The possible mechanism of the two-dimensional assembly of pentameric CRP on lipid monolayers is discussed.

Adsorption↗
PubMed2001

Gene therapy attenuates the elevated blood pressure and glucose intolerance in an insulin-resistant model of hypertension.

OBJECTIVE: Fructose feeding in male Sprague-Dawley (SD) rats results in a mild hypertension and glucose intolerance. Although the mechanism of this glucose intolerance and hypertension is not completely understood, a role for the renin-angiotensin system (RAS) has been proposed. In the current study our aim was to test the hypothesis that intervention of the RAS with a gene therapy approach would be effective in preventing the development of hypertension and glucose intolerance in this animal model. DESIGN AND METHODS: Five-day-old SD rats were administered either an empty retroviral vector (LNSV) or retroviral vector containing AT1 receptor antisense DNA (AT1R-AS). The virus (25 microl, 8 x 10(9) CFU/ml) was injected into the heart and the animals were returned to their mothers. After weaning, half the animals from each group were placed on breeder's chow or a 60% fructose diet. Indirect blood pressures (BP) were determined and an oral glucose tolerance test (OGTT) was performed when the animals had been on the respective diets for 2 months. RESULTS: Fructose-fed animals developed mild hypertension (145 +/- 3 versus 132 +/- 4 mmHg) by 6 weeks of dietary intervention. This increase in BP was prevented by AT1R-AS treatment (125 +/- 3 mmHg). At 2 months of age, fasting blood glucose was comparable among the four groups; however, the glucose excursion during the OGTT was significantly greater and more prolonged in the LNSV-treated, fructose-fed group than the other three groups. AT1R-AS treatment significantly prevented glucose intolerance in the fructose rat to levels observed in the controls. CONCLUSIONS: Early fructose dietary treatment results in moderate hypertension and glucose intolerance, which is prevented by a single neonatal treatment with AT1R-AS. These results suggest that the RAS is involved in the glucose intolerance associated with fructose feeding and that genetic intervention is effective in this rat model.

Animals↗
PubMed2001

Performance of triple ditch: effects of duration and periodic operation programs.

The periodic variation of operation states in triple ditch (BIO-DENITRO process) was investigated in a pilot-scale and a full-scale triple ditch treating raw wastewater containing 70% petrochemical wastewater and 30% domestic wastewater. Mathematical models describing the periodic variation of activated sludge concentrations in each ditch were proposed based on theoretical analysis and were verified in the pilot-scale and full-scale triple ditches respectively. The existence of optimal cycle time and time arrangement of periodic operation programs were demonstrated and discussed according to the mathematical models and the experimental results of the pilot-scale triple ditch operated in four different stages. Four rulers determining the duration and operation programs are suggested.

Bioreactors↗
PubMed2001

[Effect of ATXII on opening modes of myocyte sodium channel, action potential and QT intervals of ECG].

Single sodium channel currents were recorded by patch clamp technique in isolated guinea-pig ventricular myocytes, and action potentials of papillary muscle and ECG were conventionally measured. Sea anemone toxin (ATXII), a high affinity toxin to sodium channel, was used to change the sodium channel dynamics. Changes in the duration of action potentials and the QT interval of ECG depending on the channel modes were studied in order to provide hints to an understanding of pathogenesis of the long QT syndrome (a genetic disease). With the binding of ATXII to the sodium channel, the occurrence frequency and the open time constant of the "long opening" mode of single Na channels increased significantly, whereas the action potential durations, APD50 and APD90, were prolonged by 23% and 27% respectively. Following application of ATXII, on the other hand, the QT interval and the QTc, a rectified QT interval, increased by 18.6% and 18.9% respectively. The results suggest that the dynamics or modes of Na channel play an important role in determining the action potential duration and the QT interval of ECG. The dynamical changes in Na channels induced by genetic mutation may be partially responsible for the long QT syndrome.

Action Potentials↗
PubMed2001

Induction of cytochromes P450 1A1 and 1B1 by emodin in human lung adenocarcinoma cell line CL5.

Emodin (3-methyl-1,6,8-trihydroxyanthraquinone) is an active compound of many laxative herbal drugs. The present study aimed to determine the effects of emodin on cytochrome P450 (P450)-dependent monooxygenases of human lung adenocarcinoma CL5 cells. Treatment of CL5 cells with 100 microM emodin for 24 h induced benzo[a]pyrene hydroxylation, 7-ethoxyresorufin O-deethylation, and 7-ethoxycoumarin O-deethylation activities of S9 fractions. Immunoblot analysis of CL5 S9 proteins revealed that emodin induced proteins immunorelated to P450s 1A1 and 1B1. Northern blot analysis of total cellular RNA showed that emodin induced P450s 1A1 and 1B1 mRNA levels in CL5 cells. These inductive effects on P450 monooxygenase activity, protein, and mRNA were concentration- and time-dependent. Addition of emodin to CL5 cell microM S9 inhibited its 7-ethoxycoumarin O-deethylation activity. Treatment of CL5 cells with 10 microM 3-methylcholanthrene for 24 h induced monooxygenase activity and P450s 1A1 and 1B1 proteins and mRNA levels. Treatment of the lung cells with 100 microM emodin or purpurin (1,2,4-trihydroxyanthraquinone) for 24 h produced greater induction of P450s 1A1 and 1B1 mRNA than did anthraflavic acid (2,6-dihydroxyanthraquinone) or anthraquinone. The emodin treatment induced P450s 1A1 and 1B1 mRNA in human lung carcinoma NCI-H322 and breast cancer MCF-7 cells. Emodin induced P450 1A1, but not 1B1, mRNA in human hepatoma HepG2 cells. The present study demonstrates that emodin is an inducer of P450s 1A1 and 1B1 protein and mRNA in human lung adenocarcinoma CL5 cells. Modulation of P450 by emodin may be an important factor affecting metabolism and toxicity of the hydroxyanthraquinone in humans.

Adenocarcinoma↗
PubMed2001

[Insect juvenile hormone enhancing gene expression in silkworm baculovirus vector system].

5th instar silkworms were infected with recombinant baculovirus containing phytase gene or wild type BmNPV at 48 hr after ecdysis, then treated with 100 ppm Juvenile hormone. It showed that the expression level of phytase gene and polyhedrin gene per silkworm was increased by 30% and 40%, respectively. The LT50 was lengthened for more than 4 h, and the average weight of sick silkworm was increased by 10%. The results indicated that the improvement of expression efficiency of phytase gene and polyhedrin gene was mainly caused by longer time of virus replication in silkworm after the treatment of Juvenile hormone.

6-Phytase↗
PubMed2001

[Dynamic pre-concentration method of gas sample].

By utilizing the combination of frontal chromatography at ambient temperature, back-flushing and temperature gradient thermal desorption, a prototype of a dynamic pre-concentration system has been designed and evaluated. It realized the enrichment for gas samples of boiling point higher than - 103 degrees C without cryogenic facilities. To reduce the sample volume and enhance enrichment factor, the technique of momentarily non-steady state produced at the beginning of back-flushing, and the negative temperature gradient generated during thermal desorption were utilized, resulting a very sharp desorption band. Standard samples were tested to demonstrate the applicability of the method. At 10 mL sample volume, the enrichment factor was 100.

Absorption↗
PubMed2001

[Separation of two isomers in the products of condensation reaction with trimethoxybenzaldehyde as raw material and quick determination of the concentration of its intermediates by high performance liquid chromatography].

The content ratio of the two isomers in the products of condensation reaction with trimethoxybenzaldehyde as raw material is the key factor to improve the productivity of trimethoprim(TMP). A quantitative method using Nova-Pak C18 column (3.9 mm i.d. x 150 mm, 4 microns) and tetrahydrofuran(THF)-water (30:70, V/V) as mobile phase with flow rate of 1.0 mL/min and UV 320 nm detector was established. A good linearity was obtained in the concentration range of 0.01 mg.L-1-10 mg.L-1. The detectable limits of trimethoxybenzaldehyde and alpha-(3,4,5-trimethoxy-benzal)-beta-methoxy-propionitrile were 2.0 micrograms/L and 1.0 microgram/L respectively. The method has been used to analyze real samples with satisfactory results.

Benzaldehydes↗
PubMed2000

Zebrafish ftz-f1 gene has two promoters, is alternatively spliced, and is expressed in digestive organs.

Fushi-tarazu Factor-1 (FTZ-F1) is a family of nuclear receptors involved in various developmental processes. We have cloned a zebrafish FTZ-F1 gene, termed ff1, which belongs to the fetoprotein transcription factor/liver receptor homologue-1 (FTF/LRH-1) subgroup of the FTZ-F1 family. Four transcripts arise as a result of differential promoter usage and alternative splicing at the 3'-most exons. The longer transcript, form A, encodes a transcriptional activator. The shorter transcript, form B, lacks the activation domain, and hence could not activate transcription. The difference in promoter usage generates FF1 proteins with different N-terminal sequences. All four transcripts appear to be expressed in most of the adult tissues, whereas, during embryo development, the IIA form is the predominant transcript. Reverse transcriptase-PCR and in situ hybridization experiments showed that the ff1 transcript is expressed in the hypothalamus, spinal cord, mandibular arch and digestive organs, including pancreas, liver, and intestine. The expression of ff1 in the digestive organs implies its function in gut development.

Alternative Splicing↗
PubMed2000

Induction of cytochrome P-450 1A1 in human hepatoma HepG2 and lung carcinoma NCI-H322 cells by motorcycle exhaust particulate.

The effects of motorcycle exhaust particulate (MEP) on human cytochrome P-450 (P-450)-dependent monooxygenases were determined using human hepatoma cell line HepG2 and lung carcinoma cell line NCI-H322 treated with organic extracts of MEP from a two-stroke engine. Gas chromatography and mass spectrometry analysis of MEP extract revealed the presence of carcinogens benzo[a]pyrene, benz[a]anthracene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[g,h,i]perylene, chrysene, and indeno[1,2,3-c,d]pyrene in the chemical mixture. Treatment with MEP extract produced concentration- and time-dependent increases of monooxygenase activity in HepG2 cells. Treatment of the cells with 100 microg/ ml MEP extract for 24 h markedly increased benzo[a]pyrene hydroxylation, 7-ethoxycoumarin, and 7-ethoxyresorufin O-deethylation activities in microsomes. Immunoblot analysis of microsomal proteins using mouse monoclonal antibody 1-12-3 against P-450 1A1 revealed that MEP extract induced a P-450-immunorelated protein in the hepatoma cells. RNA blot analysis of cellular total RNA using a human P-450 1A1 3'-end cDNA probe showed that MEP extract increased the level of a hybridizable P-450 mRNA. These P-450 1A1 inductive effects of MEP extract were similar to those from treatment with 10 microM benzo[a]pyrene or 3-methylcholanthrene (3-MC) in HepG2 cells. Treatment of lung carcinoma NCI-H322 cells with 100 microg/ml MEP extract, 10 microM benzo[a]pyrene, or 3-MC resulted in induction of monooxygenase activity, protein, and mRNA of P-450 1A1, similar to the induction observed with the hepatoma cells. The present study demonstrates that MEP extract has the ability to induce human hepatic and pulmonary P-450 1A1 in the liver- and lung-derived cell lines, and the induction involves a pretranslational mechanism. Induction of the human hepatic and pulmonary P-450 1A1 in vitro may provide important information in the assessment of MEP metabolism and toxicity in humans.

Adenocarcinoma, Bronchiolo-Alveolar↗
PubMed2000

Transcriptional repression of p21((Waf1/Cip1/Sdi1)) gene by c-jun through Sp1 site.

Previously, we found that c-jun represses the tumor suppressor p21((Waf1/Cip1/Sdi1)) (p21) gene expression. In this study, we further investigated the mechanism of the inhibitory effect of c-jun on p21. After analysis of a series of deletion and point mutants of p21 promoter, we found that Sp1-3 site (-77 and -83) relative to the transcription start site played an important role for c-jun-repressing-responsive element in the p21 promoter. Both Sp1 and Sp3 transcription factors were the key factors for this event. However, the data from electrophoretic mobility shift assay indicated that c-jun did not change the Sp1 DNA-binding affinity, suggesting that additional factors may be involved in the repression of p21 by c-jun. Furthermore, c-jun could inhibit butyrate-inducing p21 gene expression through Sp1, indicating at least one common pathway whereby p21 expression is affected by c-jun and butyrate in opposing actions. Moreover, the hyperphosphorylated retinoblastoma protein (Rb) increased in c-jun expressing cells, indicating that phosphorylated Rb may play a role in regulating Sp1 to repress p21 expression. This is the first demonstration of how housekeeping factors and oncogene product counteract the function of tumor suppressor genes to control cell cycle progression.

Binding Sites↗
PubMed2000

Trimeric ring-like structure of ArsA ATPase.

ArsA protein is the soluble subunit of the Ars anion pump in the Escherichia coli membrane which extrudes arsenite or antimonite from the cytoplasm. The molecular weight of the subunit is 63 kDa. In the cell it hydrolyzes ATP, and the energy released is used by the membrane-bound subunit ArsB to transport the substrates across the membrane. We have obtained two-dimensional crystals of ArsA in the presence of arsenite on negatively-charged lipid monolayer composed of DMPS and DOPC. These crystals have been studied using electron microscopy of negatively-stained specimens followed by image processing. The projection map obtained at 2.4 nm resolution reveals a ring-like structure with threefold symmetry. Many molecular assemblies with the same ring-shape and dimensions were also seen dispersed on electron microscopy grids, prepared directly from purified ArsA protein solution. Size-exclusion chromatography of the protein sample with arsenite present revealed that the majority of the protein particles in solution have a molecular weight of about 180 kDa. Based on these experiments, we conclude that in solution the ArsA ATPase with substrate bound is mainly in a trimeric form.

Adenosine Triphosphatases↗