Biomedical subjects

S Fang

Publications and source records attributed to S Fang.

At least 19 recordsLinked to original sources

PubMed2001

The tumor autocrine motility factor receptor, gp78, is a ubiquitin protein ligase implicated in degradation from the endoplasmic reticulum.

gp78, also known as the tumor autocrine motility factor receptor, is a transmembrane protein whose expression is correlated with tumor metastasis. We establish that gp78 is a RING finger-dependent ubiquitin protein ligase (E3) of the endoplasmic reticulum (ER). Consistent with this, gp78 specifically recruits MmUBC7, a ubiquitin-conjugating enzyme (E2) implicated in ER-associated degradation (ERAD), through a region distinct from the RING finger. gp78 can target itself for proteasomal degradation in a RING finger- and MmUBC7-dependent manner. Importantly, gp78 can also mediate degradation of CD3-delta, a well-characterized ERAD substrate. In contrast, gp78 lacking an intact RING finger or its multiple membrane-spanning domains stabilizes CD3-delta. gp78 has thus been found to be an example of a mammalian cellular E3 intrinsic to the ER, suggesting a potential link between ubiquitylation, ERAD, and metastasis.

Animals↗

PubMed2001

Identification of rice black-streaked dwarf fijivirus in maize with rough dwarf disease in China.

Three virus isolates from maize with rough dwarf in different provinces in China were analyzed at the molecular level. When compared to an isolate from diseased rice plants in Hubei Province, all four isolates had identical genomic RNA electrophoretic profiles, which were composed of ten double-stranded (ds) RNAs. Full-length cDNAs of segment 10 (S10) from each of the four isolates were cloned by RT-PCR and the complete sequences were determined. Analysis of the sequences revealed that each consisted of 1801 nucleotides and contained a single open reading frame (ORF) which potentially encoded a protein with 558 amino acids. Further, the sequences showed more than 97.0% and 98.0% identity at nucleotide and amino acid levels, respectively. In addition, their identities to rice black-streaked dwarf virus S10 were significantly higher than those to maize rough dwarf virus S10. Based on these results, it is suggested that the virus which causes this maize disease in China is rice black-streaked dwarf virus.

China↗

PubMed2001

Three-dimensional imaging of embryonic mouse kidney by two-photon microscopy.

Developing mammalian embryonic kidney becomes progressively more elaborate as the ureteric bud branches into undifferentiated mesenchyme. Morphological perturbations of nephrogenesis, such as those seen in inherited renal diseases or induced in transgenic animals, require careful and often tedious documentation by multiple methodologies. We have applied a relatively quick and simple approach combining two-photon microscopy and advanced three-dimensional (3-D) imaging techniques to visualize and evaluate these complex events. As compared with laser confocal microscopy, two-photon microscopy offers superior optical sectioning deep into biological tissues, permitting analysis of large, heterogeneous, 3-D structures such as developing mouse kidney. Embryonic and newborn mouse kidneys were fluorescently labeled with lectins, phalloidin, or antibody. Three-dimensional image volumes were then collected. The resulting volume data sets were processed using a novel 3-D visualization technique. Reconstructed image volumes demonstrate the dichotomous branching of ureteric bud as it progresses from a simple, symmetrical structure into an elaborate, asymmetrical collecting system of multiple branches. Detailed morphology of in situ cysts was elucidated in a transgene-induced mouse model of polycystic kidney disease. We expect this integration of two-photon microscopy with advanced 3-D image analysis will provide a powerful tool for illuminating a variety of complex developmental processes in multiple dimensions.

Animals↗

PubMed2001

Prediction of early tolerance to enteral feeding in preterm infants by measurement of superior mesenteric artery blood flow velocity.

AIMS: To evaluate whether serial Doppler measurements of superior mesenteric artery (SMA) blood flow velocity after the first enteral feed could predict early tolerance to enteral feeding in preterm infants. METHODS: When clinicians decided to start enteral feeds, Doppler ultrasound blood flow velocity in the SMA was determined before and after a test feed of 0.5 ml milk. The number of days taken for infants to tolerate full enteral feeding (150 ml/kg/day) was recorded. RESULTS: Fourteen infants (group 1) achieved full enteral feeding within seven days. Thirty infants (group 2) took 8-30 days. There was no difference in the preprandial time averaged mean velocity (TAMV) between the groups at a median age of 3 (2-30) days. In group 1, there was a significant increase in TAMV (p<0.01) above the preprandial level at 45 and 60 minutes, but this did not occur in group 2. An increase in TAMV by more than 17% at 60 minutes has a sensitivity of 100% and a specificity of 70% for the prediction of early tolerance to enteral feeds. CONCLUSIONS: There is a significant correlation between an increase in mean SMA blood flow velocity and early tolerance of enteral feeding. Doppler measurements of SMA blood flow velocity may be useful for deciding when to feed high risk preterm infants.

Analysis of Variance↗

PubMed2001

[Prevalent forecast of kiwifruit bacterial canker caused by Pseudomonas syringae pv. actinidiae].

The prevalent analysis of kiwifruit bacterial canker for several years showed that the effective ecological factors of severe degree were the precipitation (x1) in the second and last ten-days of March, and the average temperature (x2) of January. The model was y = 2.1359 + 0.0107x1 - 0.6061x2. The main factor of the prevalence was the relative variation of ten-days average temperature and precipitation in Winter and in early Spring, and the regression equation was y = -8.127 + 22.739x - 13.254x2. The forecast effect of the equation was obviously significant after testing.

Actinidia↗

PubMed2001

[Ecological factors affecting prevalence of kiwifruit bacterial canker and bacteriostatic action of bacteriocides on Pseudomonas syringae pv. actinidae].

According to the systematic study for 5 years, this paper dealt with the ecological factors affecting prevalence of kiwifruit bacterial canker. The disease was more serious in the orchards with above 750 m in elevation, and more serious at mountain slope facing south than facing north. Incidence differed significantly among cultivars, with easy disease infection for Jinfeng, and strong disease resistance for Jinkui. The older the trees, the more the diseased plants. The one year old twigs had highest death twig incidence of diseased twigs and death twig incidence than other twigs. Among 6 tested bacteriocides, Jiaruinong and streptomycin showed best bacteriostatic effect.

Actinidia↗

PubMed2000

Effects of gas adsorption and collisions on electrical transport in single-walled carbon nanotubes

Degassing of bundles of single-walled carbon nanotubes in vacuum at 500 K is found to drive the thermoelectricpower (TEP) strongly negative, indicating that the degassed metallic tubes in a bundle are n type. The magnitude of the negative TEP indicates that important asymmetry in the electronic carbon pi bands exists near the Fermi energy. Easily measurable increases in the TEP ( approximately 5-10 &mgr;V/K) and resistivity ( 2%-10%) are observed at 500 K upon exposure to N2 and He, suggesting that even gas collisions with the nanotube wall can contribute significantly to the transport properties.

Journal Article↗

PubMed2000

Ubiquitin protein ligase activity of IAPs and their degradation in proteasomes in response to apoptotic stimuli.

To determine why proteasome inhibitors prevent thymocyte death, we examined whether proteasomes degrade anti-apoptotic molecules in cells induced to undergo apoptosis. The c-IAP1 and XIAP inhibitors of apoptosis were selectively lost in glucocorticoid- or etoposide-treated thymocytes in a proteasome-dependent manner before death. IAPs catalyzed their own ubiquitination in vitro, an activity requiring the RING domain. Overexpressed wild-type c-IAP1, but not a RING domain mutant, was spontaneously ubiquitinated and degraded, and stably expressed XIAP lacking the RING domain was relatively resistant to apoptosis-induced degradation and, correspondingly, more effective at preventing apoptosis than wild-type XIAP. Autoubiquitination and degradation of IAPs may be a key event in the apoptotic program.

Animals↗

PubMed2000

Mdm2 is a RING finger-dependent ubiquitin protein ligase for itself and p53.

Mdm2 has been shown to regulate p53 stability by targeting the p53 protein for proteasomal degradation. We now report that Mdm2 is a ubiquitin protein ligase (E3) for p53 and that its activity is dependent on its RING finger. Furthermore, we show that Mdm2 mediates its own ubiquitination in a RING finger-dependent manner, which requires no eukaryotic proteins other than ubiquitin-activating enzyme (E1) and an ubiquitin-conjugating enzyme (E2). It is apparent, therefore, that Mdm2 manifests an intrinsic capacity to mediate ubiquitination. Mutation of putative zinc coordination residues abrogated this activity, as did chelation of divalent cations. After cation chelation, the full activity could be restored by addition of zinc. We further demonstrate that the degradation of p53 and Mdm2 in cells requires additional potential zinc-coordinating residues beyond those required for the intrinsic activity of Mdm2 in vitro. Replacement of the Mdm2 RING with that of another protein (Praja1) reconstituted ubiquitination and proteasomal degradation of Mdm2. However, this RING was ineffective in ubiquitination and proteasomal targeting of p53, suggesting that there may be specificity at the level of the RING in the recognition of heterologous substrates.

Alkylation↗

PubMed2000

Novel rearranged ions observed for protonated peptides via metastable decomposition in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

C-terminal rearrangement ions [b(n-1) + H2O] (where n refers to the total number of residues of peptides) are frequently observed for peptides which contain basic amino acid(s), especially arginine, at or near their N termini in low- and high-energy collision-induced dissociation or post-source decay (PSD) spectra. Here we report a novel rearrangement, associated with PSD for serine- or threonine-containing peptides that are susceptible to C-terminal rearrangement. Based on PSD analyses of serine- or threonine-containing bradykinin and its analogs, which have been ethyl-esterified or 18O labeled at their C termini, the [b(k) + H2O] (where k denotes the position adjacent to the left of the Ser/Thr residue) ion is generally thought to be formed by the transfer of the hydroxyl moiety of a serine or threonine residue to the carbonyl group of the residue to its left accompanied by the loss of the remaining C-terminal portion of the peptide. When the Ser/Thr is at or near the C terminus, the present [b(k) + H2O] ion could be formed via two pathways, i.e., the Ser/Thr-related rearrangement and the conventional C-terminal rearrangement, which has been clearly verified by 18O labeling at the C terminus. In addition, the ions which are formally designated as [y(m)b(l) + H2O], where y(m)b(l) denotes a b-type internal ion, are also briefly described.

Bradykinin↗

PubMed2000

Development of a transgenic mouse that overexpresses a novel product of the growth hormone-releasing hormone gene.

The GH-releasing hormone (GHRH) precursor molecule contains a 30-amino acid C-terminal region that has been designated GHRH-related peptide (GHRH-RP). To begin to understand the physiological role of GHRH-RP, transgenic (Tg) mice that constituitively express this peptide were developed. To generate these mice, a transgene (SS-RP) was constructed by overlap primer extension PCR. This transgene, under the control of the mouse phosphoglycerate kinase gene, selectively expresses GHRH-RP, but not GHRH. Western blot analysis confirmed that the transgene produces GHRH-RP. Animals were evaluated for the effect of excess GHRH-RP on growth, fertility, behavior, stem cell factor (SCF) expression, and hematopoiesis. Northern blot and RT-PCR were used to demonstrate ubiquitous expression of the transgene in tissues from GHRH-RP Tg animals. These tissues also had marked overexpression of SCF messenger RNA compared with controls. Tg animals had significantly increased cell cycling for granulocyte-macrophage, erythroid, and multilineage progenitor cells. Transgenic animals did not differ from control mice in their growth, fertility, or behavior. These findings demonstrate, for the first time, that in vivo the C-terminal peptide of the pro-GHRH molecule is a biologically active peptide that is capable of stimulating the expression of SCF and hematopoiesis in vivo and suggests that GHRH-RP may play a role in normal blood cell development.

Animals↗

PubMed1999

RING fingers mediate ubiquitin-conjugating enzyme (E2)-dependent ubiquitination.

A RING finger-containing protein (AO7) that binds ubiquitin-conjugating enzymes (E2s) and is a substrate for E2-dependent ubiquitination was identified. Mutations of cation-coordinating residues within AO7's RING finger abolished ubiquitination, as did chelation of zinc. Several otherwise-unrelated RING finger proteins, including BRCA1, Siah-1, TRC8, NF-X1, kf-1, and Praja1, were assessed for their ability to facilitate E2-dependent ubiquitination. In all cases, ubiquitination was observed. The RING fingers were implicated directly in this activity through mutations of metal-coordinating residues or chelation of zinc. These findings suggest that a large number of RING finger-containing proteins, with otherwise diverse structures and functions, may play previously unappreciated roles in modulating protein levels via ubiquitination.

Amino Acid Sequence↗

PubMed1999

Antigenic epitopes of the hepatitis A virus polyprotein.

Forty-two antigenic domains were identified across the hepatitis A virus (HAV) polyprotein by using a set of 237 overlapping 20-mer synthetic peptides spanning the entire HAV polyprotein and a panel of serum samples from acutely HAV-infected patients. The term "antigenic domain" is used in this study to define a protein region spanned with consecutive overlapping immunoreactive peptides. Nineteen antigenic domains were found within the structural proteins, and 22 were found within the nonstructural proteins, with 1 domain spanning the junction of VP1 and P2A proteins. Five of these domains were considered immunodominant, as judged by both the breadth and the strength of their immunoreactivity. One domain is located within the VP2 protein at position 57-90 aa. A second domain, located at position 767-842 aa, contains the C-terminal part of the VP1 protein and the entire P2A protein. A third domain, located at position 1403-1456 aa, comprises the C-terminal part of the P2C protein and the N-terminal half of the P3A protein. The fourth domain, located at position 1500-1519 aa, includes almost the entire P3B, and the last domain, located at position 1719-1764 aa, contains the C-terminal region of the P3C protein and the N-terminal region of the P3D protein. It is interesting to note that four of the five most immunoreactive domains are derived from small HAV proteins and/or encompass protein cleavage sites separating different HAV proteins. The HAV-specific immunoreactivity of each antigenically reactive peptide was confirmed by using seven HAV seroconversion panels. Collectively, these data demonstrate that HAV structural and nonstructural proteins contain antigenic epitopes that can be efficiently modeled with short synthetic peptides.

Amino Acid Sequence↗

PubMed1999

Binding and uptake of cationic lipid:pDNA complexes by polarized airway epithelial cells.

To better understand the barriers associated with cationic lipid-mediated gene transfer to polarized epithelial cells, Fischer rat thyroid (FRT) cells and polarized normal human bronchial epithelial (NHBE) cells grown on filter supports at an air-liquid interface were used to study the binding and uptake of cationic lipid:plasmid DNA (pDNA) complexes. The efficiencies of binding and uptake of cationic lipid:pDNA complexes by these cell systems were monitored using fluorescence microscopy of fluorescently tagged lipid or pDNA probes. Fluorescent probe bound to the cell surface was differentiated from internalized probe by adding trypan blue, which quenched the fluorescence of bound but not internalized probes. For proliferating cells, binding and internalization of the cationic lipid:pDNA complexes were determined to be efficient. In contrast, little binding or internalization of the complexes was observed using polarized epithelial cells. However, after aspirating a small area of cells from the filter support, virtually all of the cells adjoining this newly formed edge bound and internalized the cationic lipid:pDNA complexes. To determine if their uptake in edge cells was related to the ability of the complexes to access the basolateral membranes of these cells, the binding and uptake of complexes was monitored in polarized NHBE cells that had been pretreated with EGTA or Ca2+-free media, strategies known to disrupt tight junctions. Cells treated in this manner bound and internalized cationic lipid:pDNA complexes efficiently and also expressed significant levels of transgene product. Control cells with intact tight junctions neither bound complexes nor expressed significant transgene product. These data confirm and extend earlier observations that the polarized apical membranes of airway epithelial cells are resistant to transfection by lipid:pDNA complexes. Further, in contrast to previous studies that have shown the entry step of complexes is not an important barrier for COS and HeLa cells, binding and entry of complexes in polarized NHBE cells appear to be rate limiting. These findings suggest that strategies designed to open the tight junctions of polarized epithelial cells may improve gene delivery to these cells for diseases such as cystic fibrosis (CF).

Alkaline Phosphatase↗

PubMed1999

Phylogeny and evolution of the Drosophila nasuta subgroup based on mitochondrial ND4 and ND4L gene sequences.

The sequences of the mitochondrial ND4 gene (1339 bp) and the ND4L gene (290 bp) were determined for all the 14 extant taxa of the Drosophila nasuta subgroup. The average A + T content of ND4 genes is 76.5% and that of ND4L genes is 83.5%. A total of 114 variable sites were scored. The ND4 gene sequence divergence ranged from 0 to 5.4% within the subgroup. The substitution rate of the ND4 gene is about 1.25% per million years. The base substitution of the genes is strongly transition biased. Neighbor-joining and parsimony were used to construct a phylogeny based on the resultant sequence data set. According to these trees, five distinct mtDNA clades can be identified. D. niveifrons represents the most diverged lineage. D. sulfurigaster bilimbata and D. kepulauana form two independent lineages. The other two clades are the kohkoa complex and the albomicans complex. The kohkoa complex consists of D. sulfurigaster sulfurigaster, D. pulaua, D. kohkoa, and Taxon-F. The albomicans complex can be divided into two groups: D. nasuta, D. sulfurigaster neonasuta, D. sulfurigaster albostrigata, and D. albomicans from Chiangmai form one group; and D. pallidifrons, Taxon-I, Taxon-J, and D. albomicans from China form the other group. High genetic differentiation was found among D. albomicans populations. Based on our phylogenetic results, we hypothesize that D. niveifrons diverged first from the D. nasuta subgroup in Papua New Guinea about 3.5 Mya. The ancestral population spread to the north and when it reached Borneo, it diversified sequentially into the kohkoa complex, D. s. bilimbata, and D. kepulauana. About 1 Mya, another radiation occurred when the ancestral populations reached the Indo-China Peninsula, forming the albomicans complex. Discrepancy between morphological groupings and phylogenetic results suggests that the male morphological traits may not be orthologous.

Animals↗

PubMed1999

NOS gene transfer inhibits expression of cell cycle regulatory molecules in vascular smooth muscle cells.

The mechanisms of nitric oxide (NO)-mediated inhibition of vascular smooth muscle (VSM) cell proliferation are still obscure. Cyclins A and E in association with cyclin-dependent kinase 2 (cdk2) serve as positive regulators for mammalian cell cycle progression through the G1/S checkpoint of the cell cycle and subsequent cell proliferation. Therefore, we have tested the effect of adenovirus-mediated transfection of the endothelial nitric oxide synthase (eNOS) gene into guinea pig coronary VSM cells on platelet-derived growth factor (BB homodimer) (PDGF-BB)-stimulated cell proliferation and the expression of cell cycle regulatory molecules. Transfection of the eNOS gene (eNOS) into VSM cells significantly inhibited (P < 0.05) [3H]thymidine incorporation into the DNA in response to PDGF-BB stimulation compared with lacZ-transfected control cells. The eNOS transfer significantly inhibited (P < 0.05) PDGF-BB-induced proliferating cell nuclear antigen (PCNA) and cyclin A expression in VSM cells compared with cells transfected with the control vector. The time course of cyclin E expression in response to PDGF-BB stimulation was delayed in eNOS-transfected cells. Levels of cyclin-dependent kinase inhibitors p21 and p27 were not significantly affected by eNOS transfer. eNOS transfer did not decrease PDGF-beta receptor number, affinity, and autophosphorylation measured by radioreceptor assay and Western analysis. These results suggest that inhibition of PDGF-stimulated expression of cyclin A, cyclin E, and PCNA is the target of NO action. These findings could explain, at least in part, NO-mediated inhibition of VSM cell proliferation.

Adenoviridae↗

PubMed1999

Enhanced recovery of injury-caused downregulation of paxillin protein by eNOS gene expression in rat carotid artery. Mechanism of NO inhibition of intimal hyperplasia?

Injury-caused dedifferentiation accompanied by proliferation and migration of smooth muscle cells (SMCs) is an important process in the development of the neointima. Nitric oxide (NO) stimulates differentiation and inhibits proliferation and migration of SMCs. Paxillin has been found to play an important role in cell differentiation, and its phosphorylation is regulated by NO in cultured SMCs. However, the regulation of paxillin by NO in the injured artery has not been investigated. Therefore, the aim of this study was to study the effects of in vivo endothelial NO synthase (eNOS) gene transfection on paxillin expression and intimal hyperplasia. A catheter balloon-denuded rat carotid artery was transfected in vivo with the replication-deficient adenovirus Ad5/RSVeNOS or with Ad5/RSVLacZ as the control. Transfected eNOS gene expression was determined by immunostaining, Western blot analysis, and citrulline assay. The expression of paxillin and its associated proteins was determined in injured arteries by Western blot analysis. The area of the intima and the ratio of intima to media were examined on cross sections by morphometry. The data showed that the expression of paxillin was significantly downregulated after injury. eNOS gene transfer showed no effect on paxillin downregulation 2 days after injury but significantly enhanced the recovery of paxillin protein 5 days and 2 weeks after injury. Vinculin, a paxillin-binding protein, was not altered by vascular injury or by eNOS gene transfer. eNOS gene transfer significantly inhibited intimal hyperplasia for up to 4 weeks. These results suggest that NO inhibition of intimal hyperplasia may be mediated by enhancing the recovery of injury-caused downregulation of paxillin.

Adenoviridae↗

PubMed1999

[Repair of soft tissue defect of heel with myocutaneous flap of flexor digitorum brevis].

OBJECTIVE: To investigate the repairing method of soft tissue defect of heel, pedicled island myocutaneous flap of flexor digitorum brevis was designed. METHODS: From 1984 to 1997, 26 cases with soft tissue defect of heel were adopted in the clinical trial. Among them, the were 18 males, 8 females and the age ranged from 15 to 60 years old. The area of wound ranged from 2.5 cm x 1.5 cm to 8.0 cm x 6.0 cm. RESULTS: After operation, all of the flaps survived. They were followed up for 9 to 72 months. All of the flaps had primary healing except in one there was infection of peripheral of the flap. The contour of heel was satisfactory the sensation of flap was good and the weight-bearing function was also successful. CONCLUSION: It was concluded that the myocutaneous flap of flexor digitorum brevis could be used to repair the soft tissue defect of heel because of its nearby position, hidden location, good recovery of skin sensation and weight-bearing function, Besides, the procedure of this operation was simple and the anti-inflammatory potential of the flap was high. However, Because of the limited donor area, the pre-operative design was important.

Adolescent↗