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D Sun

Publications and source records attributed to D Sun.

At least 73 records · Page 4Linked to original sources
PubMed2001

Effect of estrogen on flow-induced dilation in NO deficiency: role of prostaglandins and EDHF.

To investigate the role of estrogen in flow-induced dilation (FiD) in nitric oxide (NO) deficiency, FiD was examined in isolated gracilis arterioles of ovariectomized (OVX) and OVX rats with estrogen replacement (OVE). Both groups of rats were treated chronically with N(omega)-nitro-L-arginine methyl ester. Plasma concentration of NO(2)/NO(3) was reduced in both groups. Plasma concentration of estradiol was lower in OVX than in OVE rats. FiD was similar in vessels of the two groups; calculated wall shear stress and basal tone were significantly greater in OVX vs. OVE rats. Indomethacin did not affect FiD in vessels from OVE rats but abolished dilation in vessels from OVX rats. Valeryl salicylate or NS-398 inhibited FiD by approximately 50%, whereas their simultaneous administration eliminated the response in arterioles from OVX rats. In vessels from OVE rats, miconazole or charybdotoxin eliminated FiD. Thus, in NO deficiency, prostaglandins derived from both cyclooxygenase isoforms mediate FiD in gracilis arterioles of OVX rats. Estrogen replacement switches the mediation, showing dependence on endothelium-derived hyperpolarizing factor in the arterioles of OVE rats.

Animals↗
PubMed2001

Stimulation of Na-K-2Cl cotransporter in neurons by activation of Non-NMDA ionotropic receptor and group-I mGluRs.

In a previous study, we found that Na(+)-K(+)-2Cl(-) cotransporter in immature cortical neurons was stimulated by activation of the ionotropic N-methyl-D-aspartate (NMDA) glutamate receptor in a Ca(2+)-dependent manner. In this report, we investigated whether the Na(+)-K(+)-2Cl(-) cotransporter in immature cortical neurons is stimulated by non-NMDA glutamate receptor-mediated signaling pathways. Expression of the Na(+)-K(+)-2Cl(-) cotransporter and metabotropic glutamate receptors (mGluR1 and 5) was detected in cortical neurons via immunoblotting and immunofluorescence staining. Significant stimulation of cotransporter activity was observed in the presence of both trans-(+/-)-1-aminocyclopentane-trans-1,3-dicarboxylic acid (trans-ACPD) (10 microM), a metabotropic glutamate receptor (mGluR) agonist, and (RS)-3,5-dihydroxyphenylglycine (DHPG) (20 microM), a selective group-I mGluR agonist. Both trans-ACPD and DHPG-mediated effects on the cotransporter were eradicated by bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid-AM, a Ca(2+) chelator. In addition, DHPG-induced stimulation of the cotransporter activity was inhibited in the presence of mGluRs antagonist (RS)-1-aminoindan-1,5-dicarboxylic acid (AIDA) (1 mM) and also with selective mGluR1 antagonist 7-(hydroxyimino)cyclopropa[b]chromen-1a-carboxylate ethyl ester (CPCCOEt) (100 microM). A DHPG-induced rise in intracellular Ca(2+) in cortical neurons was detected with Fura-2. Moreover, DHPG-mediated stimulation of the cotransporter was abolished by inhibition of Ca(2+)/CaM kinase II. Interestingly, the cotransporter activity was increased by activation of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor. These results suggest that the Na(+)-K(+)-2Cl(-) cotransporter in immature cortical neurons is stimulated by group-I mGluR- and AMPA-mediated signal transduction pathways. The effects are dependent on a rise of intracellular Ca(2+).

Action Potentials↗
PubMed2001

Drug inhibition of Gly-Sar uptake and hPepT1 localization using hPepT1-GFP fusion protein.

An hPepT1-GFP fusion construct was made to study drug inhibition of dipeptide uptake and apical, basolateral, or subcellular hPepT1 localization. The hPepT1 stop codon was mutated by polymerase chain reaction and was subsequently cloned into the pEGFP-N1 vector. The hPepT1-GFP fusion construct was then transfected into Caco-2 and HeLa cells, and drug inhibition was studied by inhibiting 3H-Gly-Sar uptake. Western blot analysis was used to determine hPepT1-GFP expression levels and confocal microscopy was used to examine the localization. Both anti-hPepT1 antibody and anti-GFP antibody recognized a 120-kd hPepT1-GFP fusion protein in the transfected cells. The 3H-Gly-Sar uptake in transfected HeLa cells was enhanced more than 20 times compared with the control. Valacyclovir (5 mmol/L) was able to completely inhibit 3H-Gly-Sar uptake in these transfected cells. Confocal microscopy showed that the hPepT1-GFP mainly localized in the Caco-2 cell apical membrane, but was present throughout the entire HeLa cell membranes. The hPepT1-GFP fusion protein was not found in either early endosome or lysosome of Caco-2 cells under normal conditions; however, it was detected in some subsets of lysosomes and early endosomes in phorbol 12-myristate 13-acetate (PMA)-treated Caco-2 cells.

Acyclovir↗
PubMed2001

Characterization of the microtubule binding domain of microtubule actin crosslinking factor (MACF): identification of a novel group of microtubule associated proteins.

MACF (microtubule actin cross-linking factor) is a large, 608-kDa protein that can associate with both actin microfilaments and microtubules (MTs). Structurally, MACF can be divided into 3 domains: an N-terminal domain that contains both a calponin type actin-binding domain and a plakin domain; a rod domain that is composed of 23 dystrophin-like spectrin repeats; and a C-terminal domain that includes two EF-hand calcium-binding motifs, as well as a region that is homologous to two related proteins, GAR22 and Gas2. We have previously demonstrated that the C-terminal domain of MACF binds to MTs, although no homology was observed between this domain and other known microtubule-binding proteins. In this report, we describe the characterization of this microtubule-binding domain of MACF by transient transfection studies and in vitro binding assays. We found that the C-terminus of MACF contains at least two microtubule-binding regions, a GAR domain and a domain containing glycine-serine-arginine (GSR) repeats. In transfected cells, the GAR domain bound to and partially stabilized MTs to depolymerization by nocodazole. The GSR-containing domain caused MTs to form bundles that are still sensitive to nocodazole-induced depolymerization. When present together, these two domains acted in concert to bundle MTs and render them stable to nocodazole treatment. Recently, a study has shown that the N-terminal half of the plakin domain (called the M1 domain) of MACF also binds MTs. We therefore examined the microtubule binding ability of the M1 domain in the context of the entire plakin domain with and without the remaining N-terminal regions of two different MACF isoforms. Interestingly, in the presence of the surrounding sequences, the M1 domain did not bind MTs. In addition to MACF, cDNA sequences encoding the GAR and GSR-containing domains are also found in the partial human EST clone KIAA0728, which has high sequence homology to the 3' end of the MACF cDNA; hence, we refer to it as MACF2. The C-terminal domain of mouse MACF2 was cloned and characterized. The microtubule-binding properties of MACF2 C-terminal domain are similar to that of MACF. The GAR domain was originally found in Gas 2 protein and here we show that it can associate with MTs in transfected cells. Plectin and desmoplakin have GSR-containing domains at their C-termini and we further demonstrate that the GSR-containing domain of plectin, but not desmoplakin, can bind to MTs in vivo.

Amino Acid Sequence↗
PubMed2001

Development of a rapid, quantitative method for LDL subfractionation with use of the Quantimetrix Lipoprint LDL System.

BACKGROUND: Recent evidence suggests that the presence of small, dense LDL is independently associated with increased risk of developing coronary artery disease. Current methods to subfractionate LDL are time-consuming and/or technically demanding. Therefore, we have sought the development of a less complex LDL subfractionation procedure. METHODS: LDL subfractions were separated using the Quantimetrix Lipoprint(TM) LDL System. High-resolution 3% polyacrylamide gel tubes were scanned densitometrically (610 nm) with a Helena EDC system. A computerized method to identify and quantitatively score the resolved LDL subfractions was developed. Results from the Quantimetrix method were compared using 51 plasma samples with values obtained by nondenaturing gradient gel electrophoresis (NDGGE) and nuclear magnetic resonance (NMR) spectroscopy. RESULTS: LDL subfractionation scores correlated significantly (P <0.05) with triglyceride, HDL-cholesterol, apolipoprotein B100, and LDL-cholesterol/apolipoprotein B100 (r = 0.591, -0.392, 0.454, and -0.411, respectively). For 51 samples, the Quantimetrix method classified 21 with small, 14 with intermediate, and 16 with large LDL. Of the 21 samples classified as small by Quantimetrix, 20 (95%) were classified as small (n = 18) or intermediate (n = 2) by NDGGE. All of the 16 specimens classified as large by Quantimetrix were either large (n = 14) or intermediate (n = 2) by NDGGE. LDL score was inversely correlated (r = -0.674; P <0.0001) with LDL particle size determined by NMR spectroscopy. CONCLUSIONS: A quantitative method for the assessment of LDL particle size phenotype was developed using the Quantimetrix Lipoprint LDL System. The method can be performed in less than 3 h in batch mode and is suitable for routine use in clinical laboratories.

Apolipoprotein B-100↗
PubMed2001

[Effects of the alterations of membrane shear elastic modulus and viscosity on the deformation and orientation of RBCs].

Neuraminidase can partly remove the surface charge of RBCs through a biochemical interaction; thus it can give rise to alterations in the microstructure of membrane, the shear elastic modulus (E) and the viscosity(micron) of membrane. Changing the time of treatment and the dose of neuraminidase and using a new ektacytometry that can separate deformation index DI into orientation index (DI)or and small deformation index (DI)d for RBCs in shear flow field of low viscosity, we measured (DI)d and the half time t0.5 when the DI recovered to half of the maximum in the process of relaxation for every treated sample. (DI)d and t0.5 were put respectively into the RBC membrane shear elastic modulus formula and the membrane viscosity formula which were put forward by Wen Zong-yao and Yan Zong-yi et al[1]. The rules of the alterations of E and micron were obtained. We also measured DI and (DI)or. It was found that E and micron increased greatly but DI and (DI)or decreased when the dose of neuraminidase and the time of treatment were increased. There was a contrary correlation between them. These data demonstrated that the increase of E and micron weakened the deformability and the ability of orientation of RBCs.

Animals↗
PubMed2001

Elevated expression of DNA ligase I in human cancers.

PURPOSE: Human DNA ligase I plays an essential role in DNA replication, recombination, and repair by catalyzing the formation of phosphodiester bonds between adjacent 5'-phosphoryl and 3'-hydroxyl termini at single breaks in duplex DNA molecules. DNA ligase I is responsible for the majority of DNA ligase activity present in proliferating cells among four different forms of DNA ligases, designated DNA ligase I, II, III, and IV. In this study, we were interested in comparing DNA ligase I level in human tumors versus normal tissues and in studying whether the inhibition of DNA ligase I could lead to tumor cell death. EXPERIMENTAL DESIGN: DNA ligase I level was measured by Western immunoblot assay in various human malignant tumor specimens and benign tissues obtained from patients, in peripheral blood lymphocytes obtained from healthy donors, and in human tumors grown in nude mice. We also have designed antisense oligonucleotides (ODNs) targeting the mRNA of DNA ligase I and tested whether DNA ligase I antisense ODNs could control tumor cell growth. RESULTS: The amount of DNA ligase I enzyme in malignant tumors was considerably higher than that in benign normal tissues and peripheral blood lymphocytes. The level of DNA ligase I in human tumors grown in nude mice was also very high, and the expression of DNA ligase I appears to be constitutive during in vivo tumor development. We have observed that DNA ligase I antisense ODN inhibited tumor cell growth in a dose-response manner, but nonspecific control ODNs had little effect on the growth of the same cell lines. CONCLUSIONS: The presence of DNA ligase I at higher levels in human tumors than in benign normal tissues and normal peripheral lymphocytes suggests DNA ligase I plays more of a role in proliferating cells than in resting cells. These results, together with our finding that DNA ligase I antisense ODNs can suppress tumor cell proliferation, warrant the design and testing of human DNA ligase I inhibitors as new anticancer agents.

Animals↗
PubMed2001

[Intravesical instillation of cytotoxic T-lymphocytes induced by soluble tumor antigen in the prophylaxis of recurrence of bladder cancer].

OBJECTIVE: To explore a new method for preventing the recurrence of bladder cancer. METHODS: With the adjuvant of BCG, cytotoxic T-lymphocytes (CTLs) were induced from human peripheral blood mononuclear cells (PBMC) by bladder tumor soluble antigen which was extracted through salt-infiltrating biochemical procedures from autologous bladder tumor. Intravesical instillation of CTLs together with their supernatants after culture for 8 to 10 days was performed in 37 patients with superficial bladder cancer after surgery. The effects of the treatment were evaluated by endocytoscopy, pathological examinations and ELISA assays for the levels of cytokines IL-2, TNF-alpha and IFN-gamma in the patients' serum and urine. The distribution of T-lymphocyte subpopulations was also monitored by SAP immuno-histochemical techniques. RESULTS: The proportion of CD3+ and CD4+ T-lymphocyte subpopulations increased significantly while CD8+ T-lymphocyte subpopulation was stable. The ratio of CD4+ to CD8+ increased to 1.60 +/- 0.29. The levels of cytokines IL-2, TNF-alpha and IFN-gamma in serum and urine increased significantly, especially the level of TNF-alpha in urine. Pathological studies revealed that non-specific inflammation is the major change in the bladder mucosa after therapy. Follow-up for 6-26 months showed that 1 patient havd moderate heterogeneity in bladder mucosa. The recurrence rate was 2.7%. Side effects were not found except transient lower fever or hematuria. CONCLUSION: Intravesical insitillation of CTLs could effectively prevent the recurrence of superficial bladder cancer after the improvement of immunological reactivity in the whole body.

Administration, Intravesical↗
PubMed2001

Design and synthesis of fluoroquinophenoxazines that interact with human telomeric G-quadruplexes and their biological effects.

In this study we have identified a new structural motif for a ligand with G-quadruplex interaction that results in biological effects associated with G-quadruplex-interactive compounds. Fluoroquinolones have been reported to possess weak telomerase inhibitory activity in addition to their better known bacterial gyrase poisoning. Starting with a fluoroquinobenzoxazine, which has modest potency in a human topoisomerase II assay, we have designed a more potent inhibitor of telomerase that has lost its topoisomerase II poisoning activity. This fluoroquinophenoxazine (FQP) interacts with G-quadruplex structures to inhibit the progression of Taq polymerase in a G-quadruplex polymerase stop assay. In addition, we demonstrate by 1H NMR studies that this compound interacts with telomeric G-quadruplex structures by external stacking to the G-tetrad with both the unimolecular fold-over and the parallel G-quadruplex structures. A photocleavage assay confirms the FQP interaction site, which is located off center of the external tetrad but within the loop region. Molecular modeling using simulated annealing was performed on the FQP-parallel G-quadruplex complex to determine the optimum FQP orientation and key molecular interactions with the telomeric G-quadruplex structure. On the basis of the results of these studies, two additional FQP analogues were synthesized, which were designed to test the importance of these key interactions. These analogues were evaluated in the Taq polymerase stop assay for G-quadruplex interaction. The data from this study and the biological evaluation of these three FQPs, using cytotoxicity and a sea urchin embryo system, were in accord with the predicted more potent telomeric G-quadruplex interactions of the initial lead compound and one of the analogues. On the basis of these structural and biological studies, the design of more potent and selective telomeric G-quadruplex-interactive compounds can be envisaged.

Animals↗
PubMed2001

[Assessing health state utility using the short form 36].

OBJECTIVE: To explore the methods of utility assessment using the short form 36. METHODS: The authors selected 70 people stratified by age, sex, and occupation in a city in Sichuan province, and investigated the preference of the population on each domain or item by paired comparison with other domains or items. Based on the preference, the weights of the domains and items were calculated. RESULTS: The weights of all the 8 domains and the items within 7 domains except the first domain "physical functions" were identified. The total score and the scores of the domains calculated by weighted adding method were significantly different from those calculated by simple adding method. However, only little variation of the scores of the domains between the two methods appeared. CONCLUSION: Utility calculated by weighted adding method may reflect people's preference to the health states more accurately. However, the value is limited in using weighted adding method for calculating the scores of domains.

Activities of Daily Living↗
PubMed2001

[Specific amplification of the coding sequences of BBTV III, IV, I and their application in BBTV detection].

Banana bunchy top virus disease (BBTD) is a disastrous disease in bananas, and it is spreading in the world (including China) by the banana bunchy top virus(BBTV). At present, virus-free plantlets are used to prevent BBTD in banana production, therefore, it is very important to establish a method to detect BBTV quickly, sensitively and specifically. ELISA is now popularly used to detect BBTV. The sensitivity of this method is not high enough, and needs specific antiserum, otherwise, pseudo-positive results often occur. According to DNA coding sequences of component III, IV and I of BBTV isolates from Zhangzhou, China, three pairs of primers are designed to establish a PCR method to specifically amplify parts of coding sequences of the BBTV coat protein, movement protein and replicase-association. This method is also applicable to detect BBTV of bananas or cultured banana seedlings in other regions.

Base Sequence↗
PubMed2001

[Feasibility of using short form 36 in Chinese population].

OBJECTIVE: To explore the feasibility of using the SF-36 in Chinese population. METHODS: We tested the reliability and validity of the SF-36 in 1603 rural residents, 646 urban residents, 70 high school girls, 122 patients with osteoporosis, and 150 stroke patients. RESULTS: The reliability and validity of the SF-36 in general were acceptable, however, the reliability and validity varied among different domains and in different populations. The reliability and validity of the domains "limitations in physical activities", "limitations in usual role activities because of physical health problems", "bodily pain", and "general health perception" were better than those of the domains "vitality", "limitations in social activities", "limitations in usual role activities because of emotional problems", and "mental health". CONCLUSION: The results indicate that we can use the SF-36 in Chinese population, but we have to pay attentions to the variance of the reliability and validity in different populations, especially for the interpretation of the results of the domains related to mental and social functions.

Activities of Daily Living↗
PubMed2001

[Spectral interferences of rare earth elements observed with a high resolution inductively coupled plasma atomic emission spectrometer. VI. Spectral interferences of lanthanum, terbium, holmium and erbium as matrices].

The spectral interferences of La, Tb, Ho and Er (1,000 micrograms.mL-1, separately) as matrices on the other rare earth elements (REEs, 1 or 5 micrograms.mL-1) were studied with a high resolution ICP atomic emission spectrometer with a grating of 3,600 grooves/mm. Totally 66 analytical lines of 15 REEs were selected as the prominent lines in the spectral interference studies. The overlapping profiles of analytical lines and interference line were obtained by scanning different solutions (blank, analyte, and four REE matrices, individually) within the selected wavelength windows (0.2 nm for each analytical line). Some new spectral lines of La, Tb, Ho and Er were observed. The "best" analytical lines of 14 REEs were recommended based on the Q-values and the true detection limits.

Erbium↗
PubMed2000

Epithelial-mesenchymal transformation is the mechanism for fusion of the craniofacial primordia involved in morphogenesis of the chicken lip.

We have previously demonstrated that epithelial-mesenchymal transformation (EMT) brings about TGF beta 3-induced confluence of craniofacial primordia that derive from the maxillary processes and give rise to the avian palate. The upper lip of the chick embryo forms by confluence of primordia also derived from the maxillary processes, but in this case, they fuse with the intermaxillary segment of the nasofrontal process. Here, we ask whether the bilateral epithelial seams formed when these primordia contact each other in vivo are removed by apoptosis (as formerly was believed to occur in developing palate) or by EMT. We found that, as is the case in the palate, the periderm of the two-layered embryonic epithelium begins to slough shortly before these primordia fuse, bringing the basal epithelial cells into close contact. We show by TUNEL staining and confirm by TEM that apoptosis occurs only in periderm. TEM reveals that basal epithelial cells contacting each other to form the midline seam produce numerous desmosomes with each other. Then, basement membrane begins to disappear, numerous filopodia extend from the basal surfaces of epithelial cells, the space between them enlarges, and the seam breaks apart, leaving mesenchymal cells in its wake. Transformation of the carboxyfluorescein (CCFSE)-labeled epithelial seam is demonstrated in vivo by detection of CCFSE bodies in mesenchymal cells that replace it. This demonstration of EMT in avian lip development lays important groundwork for understanding the causes of human cleft lip and analyzing the mechanism of action of growth factors, such as SHH and BMPs, that have been shown (J. A. Helms et al., 1997, Dev. Biol. 187, 25-35) to be involved in avian lip confluence.

Animals↗
PubMed2000

Conversion of methylamine dehydrogenase to a long-chain amine dehydrogenase by mutagenesis of a single residue.

Methylamine dehydrogenase (MADH) is a tryptophan tryptophylquinone (TTQ) dependent enzyme that catalyzes the oxidative deamination of primary amines. Amino acid residues of both the TTQ-bearing beta subunit and the noncatalytic alpha subunit line a substrate channel that leads from the protein surface to the enzyme active site. Phe55 of the alpha subunit is located at the opening of the active site. Conversion of alphaPhe55 to alanine dramatically alters the substrate preference of MADH. The K(m) for methylamine increases from 9 microM to 15 mM. The preferred substrates are now primary amines with chain lengths of at least seven carbons. The K(m) for 1, 10-diaminodecane is 11 microM, compared to 1.2 mM for wild-type MADH. Despite the large variation in K(m) values, k(cat) values are relatively unaffected by the mutation. Molecular modeling of substrates into the crystal structure of the enzyme active site and substrate channel provides an explanation for the dramatic changes in substrate specificity caused by this mutation of a single amino acid residue.

Alanine↗
PubMed2000

Spatio-temporal interaction with disease mapping.

Markov chain Monte Carlo methods are used to estimate mortality rates under a Bayesian hierarchical model. Spatial correlations are introduced to examine spatial effects relative to both regional and regional changes over time by groups. A special feature of the models is the inclusion of longitudinal variables which will describe temporal trends in mortality or incidences for different population groups. Disease maps are used to illustrate the role of different parameters in the model and pinpointing areas of interesting patterns. The methods are demonstrated by male cancer mortality data from the state of Missouri during 1973-1992. Of special interest will be the geographic variations in the trend of lung cancer mortality over the recent past. Marginal posterior distributions are used to examine effects due to spatial correlations and age difference in temporal trends. Numerical results from the Missouri data show that although spatial correlations exist, they do not have a large effect on the estimated mortality rates.

Aged↗
PubMed2000

Inhibition of experimental allergic encephalomyelitis in the Lewis rat by paclitaxel.

Experimental allergic encephalomyelitis (EAE), an animal model for multiple sclerosis (MS), is useful for preclinical testing for agents to be considered for treatment for this human demyelinating disease. Microtubules in lymphocytes play an important role in the cascade of human T cell activation, and paclitaxel (PTX), a microtubule stabilizer, can inhibit T cell function. A new formulation of micellar PTX, free of Cremophor and ethanol, was tested for its effect on the induction of EAE in Lewis rats. Adoptive EAE was induced with an encephalitogenic T cell line activated with guinea pig myelin basic protein (GP MBP) peptide 68-88. PTX (10 mg/kg) was administered 24 and 72 h after cell transfer. The clinical signs, fulminating in controls, were completely blocked by PTX, but mild CNS inflammation remained unaltered. A similar dose of PTX, given on days 6 and 8 to animals developing active EAE after immunization with GP MBP peptide 68-88 in complete Freund's adjuvant, greatly reduced the severity of paralysis and delayed the onset of disease by 8-9 days. Marked weight loss and severe toxicity were noted with higher and more prolonged administration. In vitro micellar PTX inhibited activation of encephalitogenic T cells by both specific antigen and mitogen. Lower doses and longer treatment programs may provide effective treatment with acceptable adverse effects with this agent in the treatment of inflammatory demyelinating disease.

Amino Acid Sequence↗