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Biomedical subjects

S Wang

Publications and source records attributed to S Wang.

At least 73 records · Page 4Linked to original sources
PubMed2004

Dynamics of single polymers in a stagnation flow induced by electrokinetics.

An electrokinetics-induced stagnation flow was created inside a microscale cross-channel. Compared to hydrodynamic-induced microfluidics, this flow system can be readily assembled and the operation is very simple due to a low pressure drop. Through image analysis, a fairly homogeneous, two-dimensional elongational flow was observed. The initial conformation of DNA molecules and residence time inside the flow field play important roles in determining the extent of DNA stretching. A coarse-grain molecular simulation agrees reasonably well with experimental observations.

Computer Simulation↗
PubMed2004

Inhibition of AKT survival pathway by a small molecule inhibitor in human endometrial cancer cells.

The PTEN (phosphatase and tensin homolog deleted on chromosome 10) tumour suppressor is mutated in 40-50% of human endometrial cancers. PTEN exerts its effects in part via inhibition of the antiapoptotic protein AKT. We demonstrate that two endometrial cancer cell lines that harbour PTEN mutations, Ishikawa and RL95-2, have high levels of phosphorylated AKT and high AKT kinase activity. Two additional endometrial cancer cell lines that express wild-type PTEN, Hec1A and KLE, have little phosphorylated AKT and minimal demonstrable AKT kinase activity. We tested a potential inhibitor of the AKT pathway, API-59CJ-OMe, in these four cell lines. We found that API-59CJ-OMe inhibits AKT kinase activity and induces apoptosis in the Ishikawa and RL95-2 cell lines with high AKT activity, but has little effect on Hec1A and KLE cells without AKT activity. API-59CJ-OMe may therefore have therapeutic potential for those endometrial cancers that harbour PTEN mutations and AKT activation.

Apoptosis↗
PubMed2004

Targeting specific neuronal populations using adeno- and lentiviral vectors: applications for imaging and studies of cell function.

We employ viral vectors to address questions related to the function of specific types of neurones in the central control of blood pressure. Adenoviral vectors (AVVs) or lentiviral vectors (LVVs) can be used to visualize specifically living GABAergic or noradrenergic (NAergic) neurones or to interfere with intracellular signalling within these cell types. Here, we review recent in vitro, in situ and in vivo applications of these vectors in the rat brainstem as performed in our laboratories. In organotypic slice cultures prepared from defined cardiovascular brainstem areas, viral vectors were used to study the electrophysiological properties, intracellular signalling and gene expression in selected neuronal phenotypes. In vivo, vectors were microinjected into brainstem nuclei to inhibit specific aspects of cell signalling by expression of dominant negative proteins, for example. Outcomes for cardiovascular control were measured either acutely in situ or chronically in vivo with radio telemetry in freely moving rats. We showed that AVVs and LVVs have distinct properties that need to be considered prior to their application. For example, LVVs can be manufactured very quickly, have no immunogenicity and can be pseudotyped to display higher tropism for neurones than glia. However, comparatively lower production yields of LVVs may limit their use for some types of applications. In contrast, AVVs require a lengthy construction period, are easy to amplify to high yields at moderate cost but may trigger an immune response when used at high titres in vivo. These features make AVVs particularly suitable for in vitro applications. As the two vector types complement each other in several ways we generated a shuttle system that simplifies transfer of transgene cassettes between the backbones of AVVs and LVVs. Thus, AVVs and LVVs are powerful experimental tools that can be used in a variety of experimental designs in vivo, in situ and in vitro.

Adenoviridae↗
PubMed2004

The BioCAT undulator beamline 18ID: a facility for biological non-crystalline diffraction and X-ray absorption spectroscopy at the Advanced Photon Source.

The 18ID undulator beamline of the Biophysics Collaborative Access Team at the Advanced Photon Source, Argonne, IL, USA, is a high-performance instrument designed for, and dedicated to, the study of partially ordered and disordered biological materials using the techniques of small-angle X-ray scattering, fiber diffraction, and X-ray absorption spectroscopy. The beamline and associated instrumentation are described in detail and examples of the representative experimental results are presented.

Academies and Institutes↗
PubMed2004

Genome-wide analysis of defense-responsive genes in bacterial blight resistance of rice mediated by the recessive R gene xa13.

Defense responses triggered by dominant and recessive disease resistance (R) genes are presumed to be regulated by different molecular mechanisms. In order to characterize the genes activated in defense responses against bacterial blight mediated by the recessive R gene xa13, two pathogen-induced subtraction cDNA libraries were constructed using the resistant rice line IRBB13--which carries xa13--and its susceptible, near-isogenic, parental line IR24. Clustering analysis of expressed sequence tags (ESTs) identified 702 unique expressed sequences as being involved in the defense responses triggered by xa13; 16% of these are new rice ESTs. These sequences define 702 genes, putatively encoding a wide range of products, including defense-responsive genes commonly involved in different host-pathogen interactions, genes that have not previously been reported to be associated with pathogen-induced defense responses, and genes (38%) with no homology to previously described functional genes. In addition, R-like genes putatively encoding nucleotide-binding site/leucine rich repeat (NBS-LRR) and LRR receptor kinase proteins were observed to be induced in the disease resistance activated by xa13. A total of 568 defense-responsive ESTs were mapped to 588 loci on the rice molecular linkage map through bioinformatic analysis. About 48% of the mapped ESTs co-localized with quantitative trait loci (QTLs) for resistance to various rice diseases, including bacterial blight, rice blast, sheath blight and yellow mottle virus. Furthermore, some defense-responsive sequences were conserved at similar locations on different chromosomes. These results reveal the complexity of xa13-mediated resistance. The information obtained in this study provides a large source of candidate genes for understanding the molecular bases of defense responses activated by recessive R genes and of quantitative disease resistance.

Chromosomes, Plant↗
PubMed2004

Objective facial paralysis grading based on Pface and eigenflow.

To provide physicians with an objective and quantitative measurement of single-sided facial paralysis, the paper presents a computer-based approach that is different from the nine existing, subjective and hand-performed international scales, such as House-Brackman. For voluntary expressions of a patient, this approach used Pface, which stems from Dface, to measure the asymmetry between two sides of the face and used eigenflow to measure the expression variations between the patient and normal subjects. The results from Pface and eigenflow were then combined by the support vector machine produce to Pdegree. A study of 25 subjects revealed that Pdegree could differentiate paralysis states (Pdegree > or = 0) and normal states (Pdegree < 0), with the ability to grade facial paralysis automatically. Moreover, the Pface of specific facial areas can be used in the supervision of the rehabilitation process.

Adolescent↗
PubMed2004

Alteration of heme-oxygenase-carbon monoxide pathway in calcified rat vascular smooth muscle cells.

OBJECTIVE: The aim of the present study was to investigate the changes in heme-oxygenase (HO)-carbon monoxide (CO)-cyclic guanosine monophosphate (cGMP) pathway in clacified rat vascular smooth muscle cells (VSMCs). METHODS: Calcification of cultured rat VSMCs was induced by incubation of VSMCs with beta-glycerophosphate. Cellular calcium content, ALP activities and (45)Ca uptake were measured. HO activity, HbCO formation and content of cGMP in VSMCs were determined. Immunocytochemistry for HO-1 expression was observed. RESULTS: In comparison of control VSMCs, the cellular calcium content, ALP activity and (45)Ca uptake in calcified VSMCs were obviously increased. Immunocytochemistry showed that HO-1 expression was weak and not well distributed in calcified cells as compared to non-calcified VSMCs, but interestingly, there was stronger staining in calcified nodules than in VSMCs. Compared with VSMCs, HO-1 activity in calcified cells decreased by 42.7% [36.4 +/- 2.8 pmol (mg Pr x h)(-1) vs 63.5 x 5.3 pmol (mg Pr x h)(-1), p < 0.01], and HbCO formation decreased by 39.2% (3.38 x 0.69 micromol/mg Pr vs 5.56 +/- 0.48 micromol/mg Pr, p < 0.05). The cGMP content in calcified VSMCs was 78.1% lower than that of non-calcified VSMCs (4.3 +/- 0.51 vs 19.6 +/- 1.2 pmol/mg, p < 0.01). CONCLUSION: The results showed that HO-CO-cGMP pathway in calcified vascular cells obviously changed, which might contribute to disturbance of vascular function.

Animals↗
PubMed2004

Efects of the pine needle abortifacient, isocupressic acid, on bovine oocyte maturation and preimplantation embryo development.

Isocupressic acid (ICA) [15-hydroxylabda-8 (17), 13E-dien-19-oic acid], a labdane diterpene acid, isolated from ponderosa pine (Pinus ponderosa), Lodgepole pine (Pinus contorta), common juniper (Juniperus communis) and Monterey cypress (Cupressus macrocarpa), induces abortion in pregnant cows when ingested primarily during the last trimester. The objective of this study was to investigate the effects of isocupressic acid on bovine oocyte maturation (in vitro maturation (IVM)-Experiment I) and preimplantation embryo development (in vitro culture (IVC)-Experiment II) using in vitro embryo production techniques and to subsequently evaluate viability and developmental competence of ICA-cultured embryos via embryo transfer to recipient heifers (Experiment III). A complete randomized block experimental design was used. In Experiment I and II, isocupressic acid was added to IVM or IVC media at 0 (TRT1, control), 1.3 (TRT2), and 2.6 microg/ml (TRT3) Results from Experiment I and II indicated that ICA did not inhibit oocyte maturation and did not adversely affect preinpiantation embryo development. Furthermore, results from Experiment II demonstrated that isocupressic acid enhanced bovine preimplantation embryo development in vitro in a dose dependent manner. Subsequently, Day 8 (Day 0 = IVF) blastocysts cultured in vitro in the medium containing 2.6 microg/ml ICA were transferred to recipient heifers and resulted in normal pregnancies as determined by ultrasound imaging. Subsequently, all but two births were normal as evaluated by post natal veterinary examination. In conclusion, ICA showed no adverse effects on oocyte maturation and preimplantation embryo development in vitro or subsequent viability in vivo using the ICA concentrations and in vitro culture parameters of this study.

Animals↗
PubMed2004

Fabrication of nano-structured porous PLLA scaffold intended for nerve tissue engineering.

Nerve tissue engineering (NTE) is one of the most promising methods to restore central nerve systems in human health care. Three-dimensional distribution and growth of cells within the porous scaffold are of clinical significance for NTE. In this study, an attempt was made to develop porous polymeric nano-fibrous scaffold using a biodegradable poly(L-lactic acid) (PLLA) for in vitro culture of nerve stem cells (NSCs). The processing of PLLA scaffold has been carried out by liquid-liquid phase separation method. The physico-chemical properties of the scaffold were fully characterized by using differential scanning calorimetry and scanning electron microscopy. These results confirmed that the prepared scaffold is highly porous and fibrous with diameters down to nanometer scale. As our nano-structured PLLA scaffold mimics natural extracellular matrix, we have intended this biodegradable scaffold as cell carrier in NTE. The in vitro performance of NSCs seeded on nano-fibrous scaffold is addressed in this study. The cell cultural tests showed that the NSCs could differentiate on the nano-structured scaffold and the scaffold acted as a positive cue to support neurite outgrowth. These results suggested that the nano-structured porous PLLA scaffold is a potential cell carrier in NTE.

Animals↗
PubMed2004

LIF receptor signaling modulates neural stem cell renewal.

Activation of the leukemia inhibitory factor (LIF) receptor has been reported to promote gliogenesis and also to support neural stem cell (NSC) renewal. To investigate this paradox, we isolated NSCs and generated neurospheres from embryonic mice either wild-type, heterozygous, or homozygous null for LIF receptor (LIFR)-beta. Exogenous LIF abrogated neurosphere formation and promoted expression of GFAP by all cells in wild-type and heterozygous cultures. LIF also stimulated a twofold increase in the number of multipotential clones generated from these cultures in comparison with those pretreated with EGF and FGF-2 (E+F) alone. In contrast, the clonogenicity of low-density cultures of LIFR knockout cells was reduced in comparison with that of wild-type cells grown in E+F and was unaffected by LIF. Thus, although LIFR signaling is not necessary for NSC self-renewal, it enhances both the clonogenicity and the expression of GFAP by these multipotential cells.

Animals↗
PubMed2004

Regressive and reactive changes in the connectivity patterns of rod and cone pathways of P23H transgenic rat retina.

We have used the P23H line 1 homozygous albino rat to study how progressive photoreceptor degeneration affects rod and cone relay pathways. We examined P23H retinas at different stages of degeneration by confocal microscopy of immunostained sections and electroretinogram (ERG) recordings. By 21 days of age in the P23H rat retina, there is already substantial loss of rods and reduction in rod bipolar dendrites along with reduction of metabotropic glutamate receptor 6 (mGluR6) and rod-associated bassoon staining. The cone pathway is relatively unaffected. By 150 days, when rods are absent from much of the retina, some rod bipolars remain and dendrites of rod and cone bipolar cells form synaptic complexes associated with cones and horizontal cell processes. These complexes include foci of mGluR6 and bassoon staining; they develop further by 270 days of age. Over the course of degeneration, beginning at 21 days, bipolar axon terminals atrophy and the inner retina undergoes further changes including a reduced and disorganized AII amacrine cell population and thinning of the inner plexiform layer. Electroretinogram (ERG) results at 23 days show reductions in a-wave amplitude, in rod and cone-associated b-waves (using a double flash paradigm) and in the amplitude of oscillatory potentials (OPs). By 38 days, rod scotopic a-wave responses and OPs are lost. B-wave amplitudes decline until 150 days, at which point they are purely cone-driven and remain stable up to 250 days. The results show that during the course of photoreceptor loss in the P23H rat, there are progressive degenerative changes, particularly in the rod relay pathway, and these are reflected in the changing ERG response patterns. Later reactive changes involving condensation of cone terminals and neurotransmitter receptors associated with rod and cone bipolar dendrites and with horizontal cell processes suggest that at this stage, there are likely to be complex changes in the relay of sensory information through the retina.

Amacrine Cells↗
PubMed2004

Quantitative comparison of placental expression of three aquaporin genes.

Three water channel proteins, aquaporins, have been shown to be expressed in the placentae of humans and sheep-AQP1, 3, 8; AQP1 is in the vasculature, whereas AQP3, 8 are in the trophoblast cells. In this study we used the sensitive and reproducible technique of real-time PCR to compare the level of expression of the mRNAs for AQP1, 3 and 8 in the ovine placenta at five stages of gestation (27, 45, 66, 100 and 140 days-where term approximately 150 days). AQP3 was quantitatively the most highly expressed AQP at 66, 100, and 140 days). At 27 days before significant trophoblast development had occurred, the only AQP present was AQP1, in the vasculature. The expression of these aquaporins underlie the high water and urea permeability of the ovine placenta in the last half of pregnancy.

Animals↗
PubMed2004

Preservation of vision following cell-based therapies in a model of retinal degenerative disease.

This study examines whether subretinal transplantation of a human retinal pigment epithelial cell line (ARPE19) or human Schwann cells early in the course of degeneration can limit the loss of visual acuity that normally occurs in dystrophic Royal College of Surgeons (RCS) rats as they age. The acuity of both transplanted groups was significantly better than controls at all ages tested, though the rescue profile of each cell type was unique. These data indicate that the transplantation of cells with very different phenotypes can be used to limit the deterioration of spatial vision in an animal model of retinal degenerative disease.

Animals↗
PubMed2004

Umbilical cord blood transplant for adult patients with severe aplastic anemia using anti-lymphocyte globulin and cyclophosphamide as conditioning therapy.

Allo-CBSCT (cord blood stem cell transplant) has been applied in six adult patients with severe aplastic anemia (SAA). Anti-lymphocyte globulin (ALG) 40 mg kg(-1) d(-1) x 3 days combined with cyclophosphamide (CTX) 20 mg kg(-1) d(-1) x 3 days constituted a lower intensive conditioning regimen. The prophylaxis of GVHD consisted of standard CsA and MTX. Patients are all male having a mean age of 26.5 years (range 22-38), and a median weight of 55.6 kg (range 52-60 kg). Cord blood searches were all conducted at Guangzhou Cord Blood Bank. Three of six patients in our study received one unit of cord blood in a procedure, whereas for another three patients, two units of cord blood (double units) were infused at the same time in a transplant protocol. The nine units of umbilical cord blood (UCB) infused contained 1.6-10.7 x 10(7) nucleated cells/kg body weight of the recipient after thawing. HLA antigens were identical in one unit, 1 antigen mismatched in seven, 2 antigens mismatched in 1. As of February 2003, after a median follow up of 20 months (range 7-50), four patients are alive and disease free. Five patients engrafted with molecular biology analyses showing donor-recipient mixed chimerism post transplant which is stable and persistent. One patient died of severe infection in the third month from transplant and another patient died in the early stage post transplant of serious aspergillus infection without evidence of engraftment.

Adult↗
PubMed2004

CMV enhancer/human PDGF-beta promoter for neuron-specific transgene expression.

Using cell-type-specific promoters to restrict expression of therapeutic genes to particular cells is an attractive approach for gene therapy, but often hindered by inefficient transcriptional activities of the promoters. Knowing the enhancer for the human cytomegalovirus (CMV) immediate-early gene improves activities of several cell-type- or tissue-type-specific promoters, we set out to investigate whether it improves neuronal transgene expression driven by a neuron-specific promoter, the platelet-derived growth factor B-chain (PDGF-beta) promoter. A hybrid promoter was constructed by appending a 380-bp fragment of the CMV enhancer 5' to the PDGF-beta promoter. The plasmid containing the promoter was complexed with polyethylenimine for in vitro and in vivo gene transfer. In cultured cells, the plasmid with the hybrid promoter significantly augmented expression of a luciferase reporter gene, providing expression levels 8- to 90-fold and 7- to 178-fold higher than those from two baseline constructs containing the PDGF-beta promoter alone and the CMV enhancer alone, respectively. In particular, the activities of the hybrid promoter in two neural cell lines were close to or higher than that of the CMV immediate-early gene enhancer/promoter, a transcriptional control element that has been considered to be the most robust one identified thus far. After stereotaxic injection into the hippocampus and striatum in rats, the hybrid promoter displayed a neuronal specificity, driving gene expression almost exclusively in neurons. Transgene expression in the brain driven by the hybrid promoter was detectable 24 h after injection, being 10-fold higher than that driven by the PDGF-beta promoter alone. The expression peaked around 5 days at 1.5 x 10(5) relative light units per brain and lasted for at least 4 weeks. This differed strikingly from the expression driven by the PDGF-beta promoter, which was no longer detectable on day 3. The new gene regulatory construct reported in this study will be useful to improve neuronal transgene expression required for gene therapy of neurological disorders and functional studies of the nervous system.

Animals↗
PubMed2004

CNS gene transfer mediated by a novel controlled release system based on DNA complexes of degradable polycation PPE-EA: a comparison with polyethylenimine/DNA complexes.

Nonviral gene delivery systems based upon polycation/plasmid DNA complexes are quickly gaining recognition as an alternative to viral gene vectors for their potential in avoiding immunogenicity and toxicity problems inherent in viral systems. We investigated in this study the feasibility of using a controlled release system based on DNA complexed with a recently developed polymeric gene carrier, polyaminoethyl propylene phosphate (PPE-EA), to achieve gene transfer in the brain. A unique feature of this gene delivery system is the biodegradability of PPE-EA, which can provide a sustained release of DNA at different rates depending on the charge ratio of the polymer to DNA. PPE-EA/DNA complexes, naked DNA, and DNA complexed with polyethylenimine (PEI), a nondegradable cationic polymer known to be an effective gene carrier, were injected intracisternally into the mouse cerebrospinal fluid. Transgene expression mediated by naked DNA was mainly detected in the brain stem, a region close to the injection site. With either PPE-EA or PEI as a carrier, higher levels of gene expression could be detected in the cerebral cortex, basal ganglia, and diencephalons. Transgene expression in the brain mediated by PPE-EA/DNA complexes at an N/P ratio of 2 persisted for at least 4 weeks, with a significant higher level than that produced by either naked plasmid DNA or PEI/DNA at the 4-week time point. Furthermore, PPE-EA displayed much lower toxicity in cultured neural cells as compared to PEI and did not cause detectable pathological changes in the central nervous system (CNS). The results established the potential of PPE-EA as a new and biocompatible gene carrier to achieve sustained gene expression in the CNS.

Animals↗
PubMed2004

Functional expression of ATM gene carried by HSV amplicon vector in vitro and in vivo.

Ataxia-telangiectasia (AT) is a human autosomal recessive disease with a pleiotropic phenotype characterized by cerebellar degeneration, immunodeficiency, premature aging, cancer predisposition, and radiation sensitivity. The gene mutated in AT, ATM (for AT-mutated), had been cloned and found to have ionizing radiation and oxidative stress-inducible kinase activity. No treatment can stop the progression of the disease. In this study, the complete open-reading frame of ATM cDNA was cloned into a Herpes simplex virus type-1 (HSV-1) amplicon vector (pTO-ATM), and the transduction of cultured AT cells was demonstrated by immunohistochemistry and Western blot analysis. Functional gene expression was evaluated by cell colony-forming assays following exposure to oxidative stress. The survival of AT cells with ATM gene transduction was about 100% higher compared to nontransduced cells after t-butyl hydroperoxide treatments. Next, the normal ATM gene expression in different regions of the rat brain was studied. Immunohistochemistry staining demonstrated weak endogenous ATM protein expression in neurons of the caudate-putamen, with significantly higher levels of expression detected in neurons in other brain regions. Exogenous ATM gene expression from pTO-ATM after viral transduction in the caudate-putamen of the adult rat was examined. At 3 days after injection of the pTO-ATM viral vector, abundant positive ATM staining of the neurons was found at the injection sites, in comparison to the controls. These data demonstrate that the relatively large ATM cDNA can be transduced and expressed in vitro and in vivo from an HSV amplicon viral vector. These data provide initial evidence that the replacement of the ATM gene into the cells of AT patients might be possible some day.

Animals↗