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Articles by Yu Sun in JoVE

 JoVE Bioengineering

Microfabricated Platforms for Mechanically Dynamic Cell Culture


JoVE 2224 12/26/2010

1Department of Mechanical and Industrial Engineering, University of Toronto, 2Institute of Biomaterials and Biomedical Engineering, University of Toronto, 3Faculty of Dentistry, University of Toronto

In this protocol, we demonstrate the fabrication of a microactuator array of vertically displaced posts on which the technology is based, and how this base technology can be modified to conduct high-throughput mechanically dynamic cell culture in both two-dimensional and three-dimensional culture paradigms.

Other articles by Yu Sun on PubMed

[Sequence Analysis for Hexon Genes of Types 3, 7 and 11 of Adenoviruses Identified in Beijing]

The objective of this study was to develop a rapid, sensitive and specific method for identifying and typing for adenovirus from clinical specimens and to learn about the viruses identified in Beijing on the molecular bases.

A New Carotid Artery Transplantation Model of Rats

To establish a murine carotid artery transplantation model for the study of the chronic rejection, 80 rats were divided into two groups, an allotransplant (ACI-Lewis) group and an isotransplant (Lewis-Lewis) group (control group). The donor carotid artery and the recipient carotid artery were anastomosed by using a polyethylene cuff (internal diameter: 0.7 mm, length: 3 mm).The pathological changes of carotid artery transplant were observed 14, 28 and 56 days after the transplantation. The results showed that the model was successfully established in 95% of the animals. The chronic rejection-associated arteriosclerosis was induced 28 days after the transplantation. The new chronic rejection model of carotid artery by using cuff technique caused fewer traumas and was easy to make. The pathological changes of the transplant mimicked the chronic rejection-associated arteriosclerosis found in human transplant.

Aerosolised Surfactant Generated by a Novel Noninvasive Apparatus Reduced Acute Lung Injury in Rats

Exogenous surfactant has been explored as a potential therapy for acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). In the present study, a nebuliser driven by oxygen lines found in the hospital was developed to deliver aerosolised porcine pulmonary surfactant (PPS). We hypothesised that aerosolised surfactant inhaled through spontaneous breathing may effectively reduce severe lung injury.

A Useful Device for Difficult Nasal Tracheal Intubation in China

[Isolation and Identification of Compounds from Marine Mangrove Plant Avicennia Marina]

To investigate the chemical constituents from Avicennia marina.

Exploring Pathways from Gene Co-expression to Network Dynamics

One of the major challenges in post-genomic research is to understand how physiological and pathological phenotypes arise from the networks or connectivity of expressed genes. In addressing this issue, we have developed two computational algorithms, CoExMiner and PathwayPro, to explore static features of gene co-expression and dynamic behaviors of gene networks. CoExMiner is based on B-spline approximation followed by the coefficient of determination (CoD) estimation for modeling gene co-expression patterns. The algorithm allows the exploration of transcriptional responses that involve coordinated expression of genes encoding proteins which work in concert in the cell. PathwayPro is based on a finite-state Markov chain model for mimicking dynamic behaviors of a transcriptional network. The algorithm allows quantitative assessment of a wide range of network responses, including susceptibility to disease, potential usefulness of a given drug, and consequences of such external stimuli as pharmacological interventions or caloric restriction. We demonstrated the applications of CoExMiner and PathwayPro by examining gene expression profiles of ligands and receptors in cancerous and non-cancerous cells and network dynamics of the leukemia-associated BCR-ABL pathway. The examinations disclosed both linear and nonlinear relationships of ligand-receptor interactions associated with cancer development, identified disease and drug targets of leukemia, and provided new insights into biology of the diseases. The analysis using these newly developed algorithms show the great usefulness of computational systems biology approaches for biological and medical research.

[Studies on Application of Ultra-filtration to Purifying Polysaccharides from Rhubarb]

To purify Rhubarb polysaccharides via micro-filtration and ultra-filtration.

Microengineered Platforms for Cell Mechanobiology

Mechanical forces play important roles in the regulation of various biological processes at the molecular and cellular level, such as gene expression, adhesion, migration, and cell fate, which are essential to the maintenance of tissue homeostasis. In this review, we discuss emerging bioengineered tools enabled by microscale technologies for studying the roles of mechanical forces in cell biology. In addition to traditional mechanobiology experimental techniques, we review recent advances of microelectromechanical systems (MEMS)-based approaches for cell mechanobiology and discuss how microengineered platforms can be used to generate in vivo-like micromechanical environment in in vitro settings for investigating cellular processes in normal and pathophysiological contexts. These capabilities also have significant implications for mechanical control of cell and tissue development and cell-based regenerative therapies.

Wogonoside Inhibits Lipopolysaccharide-induced Angiogenesis in Vitro and in Vivo Via Toll-like Receptor 4 Signal Transduction

Wogonoside, one flavonoid derived from the root of Scutellaria baicalensis Georgi, has been reported for its anti-inflammation activity; however, whether it can inhibit inflammation-induced angiogenesis is still unclear. In the present study, we evaluated the effect of wogonoside on lipopolysaccharide (LPS)-induced angiogenesis in vitro and in vivo. Wogonoside suppressed the LPS-stimulated migration and tube formation of human umbilical vein endothelial cells (HUVECs), as well as microvessel sprouting from rat aortic rings in vitro. Moreover, wogonoside also inhibited LPS-stimulated vessel growth of Chicken chorioallantoic membrane (CAM) in vivo. The mechanism revealed that wogonoside inhibited LPS-induced toll-like receptor 4 (TLR4) up-regulation and its downstream mitogen-activated protein kinases (MAPKs) activation, by decreasing the phosphorylation of c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase. The results suggest that wogonoside inhibits LPS-induced angiogenesis both in vitro and in vivo, and that it might have a therapeutic potential for the diseases associated with the development of both inflammation and angiogenesis progress.

A Comparative Study of Feature Extraction and Blind Source Separation of Independent Component Analysis (ICA) on Childhood Brain Tumour 1H Magnetic Resonance Spectra

Independent component analysis (ICA) has the potential of determining automatically the metabolite signals which make up MR spectra. However, the reliability with which this is accomplished and the optimal approach for investigating in vivo MRS have not been determined. Furthermore, the properties of ICA in brain tumour MRS with respect to dataset size and data quality have not been systematically explored. The two common techniques for applying ICA, blind source separation (BSS) and feature extraction (FE) were examined in this study using simulated data and the findings confirmed on patient data. Short echo time (TE 30 ms), low and high field (1.5 and 3 T) in vivo brain tumour MR spectra of childhood astrocytoma, ependymoma and medulloblastoma were generated by using a quantum mechanical simulator with ten metabolite and lipid components. Patient data (TE 30 ms, 1.5 T) were acquired from children with brain tumours. ICA of simulated data shows that individual metabolite components can be extracted from a set of MRS data. The BSS method generates independent components with a closer correlation to the original metabolite and lipid components than the FE method when the number of spectra in the dataset is small. The experiments also show that stable results are achieved with 300 MRS at an SNR equal to 10. The FE method is relatively insensitive to different ranges of full width at half maximum (FWHM) (from 0 to 3 Hz), whereas the BSS method degrades on increasing the range of FWHM. The peak frequency variations do not affect the results within the range of +/-0.08 ppm for the FE method, and +/-0.05 ppm for the BSS method. When the methods were applied to the patient dataset, results consistent with the synthesized experiments were obtained.

Targeted Connexin26 Ablation Arrests Postnatal Development of the Organ of Corti

Mutations in the gene coding for connexin26 (Cx26) is the most common cause of human nonsyndromic hereditary deafness. To investigate deafness mechanisms underlying Cx26 null mutations, we generated three independent lines of conditional Cx26 null mice. Cell differentiation and gross cochlear morphology at birth seemed normal. However, postnatal development of the organ of Corti was stalled as the tunnel of Corti and the Nuel's space were never opened. Cell degeneration was first observed in the Claudius cells around P8. Outer hair cell loss was initially observed around P13 at middle turn when inner hair cells were still intact. Massive cell death occurred in the middle turn thereafter and gradually spread to the basal turn, resulting in secondary degeneration of spiral ganglion neurons in the corresponding cochlear locations. These results demonstrated that Cx26 plays essential roles in postnatal maturation and homoeostasis of the organ of Corti before the onset of hearing.

Blind Intubation Device for Nasotracheal Intubation in 100 Oral and Maxillofacial Surgery Patients with Anticipated Difficult Airways: a Prospective Evaluation

The blind intubation device is a newly developed light-guided intubation device for difficult nasotracheal intubation. The aim of this study was to evaluate its performance in adult patients with anticipated difficult airways.

Scribble Interacts with Beta-catenin to Localize Synaptic Vesicles to Synapses

An understanding of how synaptic vesicles are recruited to and maintained at presynaptic compartments is required to discern the molecular mechanisms underlying presynaptic assembly and plasticity. We have previously demonstrated that cadherin-beta-catenin complexes cluster synaptic vesicles at presynaptic sites. Here we show that scribble interacts with the cadherin-beta-catenin complex to coordinate vesicle localization. Scribble and beta-catenin are colocalized at synapses and can be coimmunoprecipitated from neuronal lysates, indicating an interaction between scribble and beta-catenin at the synapse. Using an RNA interference approach, we demonstrate that scribble is important for the clustering of synaptic vesicles at synapses. Indeed, in scribble knockdown cells, there is a diffuse distribution of synaptic vesicles along the axon, and a deficit in vesicle recycling. Despite this, synapse number and the distribution of the presynaptic active zone protein, bassoon, remain unchanged. These effects largely phenocopy those observed after ablation of beta-catenin. In addition, we show that loss of beta-catenin disrupts scribble localization in primary neurons but that the localization of beta-catenin is not dependent on scribble. Our data supports a model by which scribble functions downstream of beta-catenin to cluster synaptic vesicles at developing synapses.

Incidence of Thromboembolic Complications in Lumbar Spinal Surgery in 1,111 Patients

Deep venous thrombosis (DVT) and pulmonary embolism (PE) cause significant morbidity and mortality in orthopaedic surgical practice, although the incidence following surgery to the lumbosacral spine is less than following lower limb surgery. Our objective was to compare our rate of thromboembolic complications with those published elsewhere and investigate whether the adoption of additional pharmacological measures reduced the incidence of clinically evident DVT and PE. This retrospective study was undertaken to investigate the incidence of DVT/PE during the 10 years from 1 January 1985 to 31 December 1994, and then to assess the effectiveness of an anticoagulant policy introduced during 1995 using low dose aspirin or LMH in high risk cases. All records for spinal operations were reviewed for thrombo-embolic complications by reference to the Scottish Morbidity Record form SMR1. To ensure that all patients were compliant with the policy, data for the whole of 1995 was omitted and the period 1 January 1996 to 31 December 2003 was taken to assess its effectiveness. Surgery was done with the patient in the kneeling, seated prone position which leaves the abdomen free and avoids venous kinking in the legs. Records of a total of 1,111 lumbar spine operations were performed from 1 January 1985 to 31 December 2004 were reviewed. The overall incidence of thrombo-embolic complications was 0.29%. A total of 697 operations were performed from 1 January 1985 to 31 December 1994 with two cases of DVT and no cases of PE giving thromboembolic complication rate of 0.29%. During the period 1 January 1996 to 31 December 2003, 414 operations resulted in one case of DVT and no cases of PE, a rate of 0.24%. The incidence of symptomatic thrombo-embolic complications in lumbar spinal surgery is low in the kneeling, seated prone operating position, whether or not anticoagulation is used.

Exogenous Porcine Surfactants Increase the Infiltration of Leukocytes in the Lung of Rats

Several studies have investigated the influence of exogenous surfactants on inflammatory response in the lung, however results reported about effects of surfactants on the lung infiltration of leukocytes are controversial. Our previous study noticed that treatment of porcine surfactant (PS) significantly increased the lung infiltration of leukocytes in rats with acute lung injury (ALI). The objective of this study was to verify the effect of exogenous PS on the lung infiltration of leukocytes in vivo and investigate the possible mechanisms involved in vitro.

[Gonadotropin-releasing Hormone Antagonist Protocol in Patients with Risk of Poor Response to Ovarian Stimulation in IVF-ET]

To evaluate the application of gonadotrophin-releasing hormone antagonist (GnRH-ant) in patients with risk of poor response to controlled ovarian stimulation in IVF-ET.

Use of Serum Homocysteine to Predict Stroke, Coronary Heart Disease and Death in Ethnic Chinese. 12-year Prospective Cohort Study

Prospective data about the association between serum total homocysteine (Hcy) and vascular disease in Asia is limited because few investigations have evaluated the cutpoint of Hcy for predicting the risk of vascular disease and death.

Cortical Interactive Network During Mental Rotation of Chinese Character

Mental rotation (MR) of Chinese characters has been proposed to employ distinct strategies depending on task difficulty. Cognitive process in MR is associated with multi-component neural networks, and elucidation of specific cortical interactions taking place during MR will assist understanding of the cognitive processes involved. In this study, we investigated cortical interactive networks involved in Chinese character MR tasks of different difficulties. Scalp electroencephalogram (EEG) signals were recorded from nine subjects (male/female=6/3) during MR of a Chinese character presented at different orientations (0 degrees, +/-60 degrees, +/-120 degrees and 180 degrees). Partial directed coherence (PDC) analysis based on multivariate Granger causality (GC) was used to assess cortical interactions. At +/-60 degrees and +/-120 degrees, lateral interactions from right to left counterparts were found in both the parietal and motor-related areas, and they were enhanced with the increase of rotation angle. The main interactions between parietal and motor-related areas showed feedforward at rotations of +/-60 degrees and +/-120 degrees, while feedback interactions appeared at rotations of +/-120 degrees. However, at 180 degrees of rotation, neither lateral interactions within motor-related areas nor feedback interactions from motor-related to parietal areas were found. These findings show that during MR of Chinese character (1) cortical interactive networks change according to task difficulty, and (2) the right hemisphere plays an initiating role in bilateral cortical activation.

Integrating Polyurethane Culture Substrates into Poly(dimethylsiloxane) Microdevices

Poly(dimethylsiloxane) (PDMS)-based microdevices have enabled rapid, high-throughput assessment of cellular response to precisely controlled microenvironmental stimuli, including chemical, matrix and mechanical factors. However, the use of PDMS as a culture substrate precludes long-term culture and may significantly impact cell response. Here we describe a method to integrate polyurethane (PU), a well-studied and clinically relevant biomaterial, into the PDMS multilayer microfabrication process, enabling the exploration of long-term cellular response on alternative substrates in microdevices. To demonstrate the utility of these hybrid microdevices for cell culture, we compared initial cell adhesion, cell spreading, and maintenance of protein patterns on PU and PDMS substrates. Initial cell adhesion and cell spreading after three days were comparable between collagen-coated PDMS and PU substrates (with or without collagen coating), but significantly lower on native PDMS substrates. However, for longer culture durations (> or = 6 days), cell spreading and protein adhesion on PU substrates was significantly better than that on PDMS substrates, and comparable to that on tissue culture-treated polystyrene. Thus, the use of a generic polyurethane substrate in microdevices enables longer-term cell culture than is possible with PDMS substrates. More generally, this technique can improve the impact and applicability of microdevice-based research by facilitating the use of alternate, relevant biomaterials while maintaining the advantages of using PDMS for microdevice fabrication.

Human Bocavirus Infections Are Common in Beijing Population Indicated by Sero-antibody Prevalence Analysis

Human bocavirus (HBoV) is a newly identified human parvovirus that was originally detected in the respiratory secretions of children with respiratory infections. This study aimed to learn about the importance of HBoV infections by revealing the prevalence of serum antibodies against HBoV in Beijing population.

Effects and Mechanism of Oridonin on Pulmonary Hypertension Induced by Chronic Hypoxia-hypercapnia in Rats

Pulmonary arterial hypertension (PAH) is characterized by suppressing apoptosis and enhancing cell proliferation in the vascular wall. Inducing pulmonary artery smooth muscle cells (PASMC) apoptosis had been regarded as a therapeutic approach for PAH. Oridonin can cause apoptosis in many cell lines, while little has been done to evaluate its effect on PASMC.

[Profile of Serum Soluble Endoglin in Pregnant Women with Severe Pre-eclampsia and Eclampsia]

To discuss the serum endoglin expression in severe pre-eclampsia and eclampsia women and their relationships.

Microfabricated Glass Devices for Rapid Single Cell Immobilization in Mouse Zygote Microinjection

This paper presents the design and microfabrication of a vacuum-based cell holding device for single-cell immobilization and the use of the device in mouse zygote microinjection. The device contains many through-holes, constructed via two-sided glass wet etching and polydimethylsiloxane (PDMS)-glass bonding. Experimental results of mouse zygote immobilization and microinjection demonstrate that the device is effective for rapid cell immobilization and does not produce negative effect on embryonic development.

Early Cortical Connective Network Relating to Audiovisual Stimulation by Partial Directed Coherence Analysis

Cross-modal sensory integration between the auditory and visual systems has long been a topic of interest in cognitive neuroscience, but spatiotemporal cortical interdependence under audiovisual stimulation is still poorly understood. In this letter, parametric spectral analysis, specifically partial directed coherence (PDC), is used to study the cortical connective causal-interdependence network under audiovisual cognitive processes within the time window of the N1 component, i.e., 96--145 ms poststimulus in the event-related potential (ERP). A total of 13 subjects (age: 22.9+/-0.8 years; male/female = 9/4) performed an audiovisual recognition task. Multichannel EEG signals were analyzed using an adaptive multivariate autoregressive model to identify causal influences between different cortical areas. Three different cortical connective networks were detected, and by comparing the cortical PDC network in the early cognitive process of audiovisual (VA) perception with that in unimodal situations (V or A), it was found that the left temporal cortex (TC) might be responsible for audiovisual integration. Such electrophysiological findings were highly consistent with previous functional neuroimaging results. Moreover, the results presented here show the dynamics of lateral causal influences during audiovisual integration.

Retrograde Conversion of Oral to Nasotracheal Intubation Using a Suction Catheter in a Pediatric Patient

Airway Rescue with an Esophageal Airway in a Child with a Huge Venous Malformation in Oral and Maxillofacial Region--a Case Report

Connexin30 Null and Conditional Connexin26 Null Mice Display Distinct Pattern and Time Course of Cellular Degeneration in the Cochlea

Mutations in connexin26 (Cx26) and Cx30 are the most common cause of nonsyndromic inherited deafness in humans. To understand the underlying molecular mechanisms, we investigated the pattern and time course of cellular degeneration in the cochlea of conditional Cx26 (cCx26) null and Cx30 null mice. In cCx26 null mice, initial degeneration was observed around postnatal day 14 in outer hair cells (OHCs) and supporting cells surrounding the OHCs. All cells in the middle turn organ of Corti were lost 1 month after birth, and degeneration gradually spread to the basal and apical turns. Most spiral ganglion (SG) neurons in the middle and basal turns disappeared in the first 3 months, whereas significant amounts of apical SG neurons survived. In the cochlea of Cx30 null mice, survival of most inner HCs, supporting cells, and SG neurons was observed for up to 18 months. The most severe degeneration was found in apical SG neurons and OHCs. OHC loss followed a slow time course and a base to apex gradient. Gross structures of the endolymphatic space and stria vascularis observed at the light microscope level were unchanged in either Cx null mouse models. This study revealed that cellular degeneration in the cochlea of cCx26 null mice was dramatically more rapid and widespread than that observed in Cx30 null mice. The radically different pathogenesis processes displayed by cCx26 and Cx30 null mice suggest heterogeneous underlying deafness mechanisms, despite co-assembly of Cx26 and Cx30 in forming gap junctions in the cochlea.

Treatment with Immunosuppressants FTY720 and Tacrolimus Promotes Functional Recovery After Spinal Cord Injury in Rats

Spinal cord injury (SCI) occurs frequently and is a leading cause of permanent disability in young adults. Many immune inhibitors including tacrolimus (FK506) are shown to be helpful in the regeneration of neural tissue following spinal cord injury. FTY720 belongs to a new class of immunosuppressants. The combination of FTY720 and tacrolimus has been reported to elicit synergistic immunosuppresive effects in rat allograft models without causing critical adverse effects. This study was to determine whether the combination of FTY720 and tacrolimus is superior to FTY720 or tacrolimus alone in the treatment of SCI. Forty-eight rats were subjected to a weight-drop contusion at the tenth thoracic level (a 10-g rod dropped from a height of 25 mm). At 30 min after the operation, they were randomly divided into four groups and received treatment with either FTY720 (0.5 mg/kg), tacrolimus (0.5 mg/kg), FTY720 + tacrolimus (0.5 mg/kg and 0.5 mg/kg respectively) or saline via gavage. Functional recovery was evaluated during 42 days after SCI via open-field test, inclined plane test, footprint analysis, somatosensory evoked potentials (SSEPs), and electron microscopic analysis. Rats from three treatment groups showed significantly better locomotor functional outcomes, higher SSEP amplitude, shorter SSEP latency, and milder pathological changes compared with those of control group. Moreover, rats treated with a combination of FTY720 and tacrolimus demonstrated significantly greater functional recovery by day 14 after SCI than those treated with either FTY720 or tacrolimus alone. These results suggest that the combination of FTY720 and tacrolimus could be a potentially effective therapeutic strategy to treat SCI.

A Facile Access to a Novel Bidentate Enantiomerically Pure P,N-donor Ligand

The synthesis of a new chiral P,N-donor ligand containing a phosphite and a pyrazole site and its coordination chemistry with transition metals are described.

A Monolithic Polymeric Microdevice for PH-responsive Drug Delivery

A drug-delivery microdevice integrating pH-responsive nano-hydrogel particles functioning as intelligent nano valves is described. The polymeric microdevices are monolithic without requiring peripheral control hardware or additional components for controlling drug-release rates. pH-responsive nanoparticles were synthesized and embedded into a composite membrane. The resulting pH-responsive composite membranes were integrated with PDMS micro reservoirs via a room-temperature transfer bonding technique to form the proof-of-concept microdevices. In vitro release characterization of the microdevices was conducted in which the release rate of Vitamin B(12) (VB(12)) as a model drug increased dramatically when the local pH value was decreased from 7.4 to 4. This device concept can serve as a platform technology for intelligent drug delivery in response to various in vivo environmental signals.

Dynamic Analysis of Bone Marrow Mesenchymal Stem Cells Migrating to Pancreatic Islets Using Coculture Microfluidic Chips: An Accelerated Migrating Rate and Better Survival of Pancreatic Islets Were Revealed

A coculture microfluidic chip had been fabricated to investigate the dynamic process of bone marrow mesenchymal stem cells migrating to the pancreatic islets in vitro.

Phosphorylation of Caveolin-1 Regulates Oxidant-induced Pulmonary Vascular Permeability Via Paracellular and Transcellular Pathways

Oxidants are important signaling molecules known to increase endothelial permeability, although the mechanisms underlying permeability regulation are not clear.

Brassinosteroid Signal Transduction from Cell-surface Receptor Kinases to Nuclear Transcription Factors

Brassinosteroid (BR) regulates gene expression and plant development through a receptor kinase-mediated signal transduction pathway. Despite the identification of many components of this pathway, it remains unclear how the BR signal is transduced from the cell surface to the nucleus. Here we describe a complete BR signalling pathway by elucidating key missing steps. We show that phosphorylation of BSK1 (BR-signalling kinase 1) by the BR receptor kinase BRI1 (BR-insensitive 1) promotes BSK1 binding to the BSU1 (BRI1 suppressor 1) phosphatase, and BSU1 inactivates the GSK3-like kinase BIN2 (BR-insensitive 2) by dephosphorylating a conserved phospho-tyrosine residue (pTyr 200). Mutations that affect phosphorylation/dephosphorylation of BIN2 pTyr200 (bin2-1, bin2-Y200F and quadruple loss-of-function of BSU1-related phosphatases) support an essential role for BSU1-mediated BIN2 dephosphorylation in BR-dependent plant growth. These results demonstrate direct sequential BR activation of BRI1, BSK1 and BSU1, and inactivation of BIN2, leading to accumulation of unphosphorylated BZR (brassinazole resistant) transcription factors in the nucleus. This study establishes a fully connected BR signalling pathway and provides new insights into the mechanism of GSK3 regulation.

[The Mechanism of Counter-modulatory Effect of Migration Inhibitory Factor on Anti-inflammatory Action of Glucocorticoids]

Plasma Level of Soluble C-Met is Tightly Associated with the Clinical Risk of Preeclampsia

The objective of the study was to examine the relevance of the soluble form c-Met (sMet) with the clinical risk for severe preeclampsia.

Elevated CO2 Changes Interspecific Competition Among Three Species of Wheat Aphids: Sitobion Avenae, Rhopalosiphum Padi, and Schizaphis Graminum

Effects of elevated CO2 (twice ambient) on the interspecific competition among three species of wheat aphids (Sitobion avenae, Rhopalosiphum padi, and Schizaphis graminum) and on wheat-aphid interactions were studied. Wheat plants had higher biomass and yield and lower water and nitrogen content of grain when grown under elevated CO2 than under ambient CO2; levels of condensed tannins, total phenols, and total nonstructural carbohydrates were also higher in wheat ears under elevated CO2. Compared with ambient CO2, elevated CO2 increased the abundance of R. padi when introduced solely but reduced its abundance when S. avenae was also present. The spatial distribution of wheat aphids was apparently influenced by CO2 levels, with significantly more S. avenae on ears and a more even distribution of R. padi on wheat plants under elevated CO2 versus ambient CO2. Elevated CO2 did not affect the abundance and spatial distribution of S. graminus when inoculated solely. Moreover, when S. avenae was present with either R. padi or S. graminum, spatial niche overlap was significantly decreased with elevated CO2. When three species co-occurred, elevated CO2 reduced spatial niche overlap between S. avenae and S. graminum and between R. padi and S. graminum. Our results suggest that increases in atmospheric CO2 would alleviate interspecific competition for these cases, which would accentuate the abundance of and the damage caused by these wheat aphids.

[Effects of Elevated O3 Concentration on Anti-oxidative Enzyme Activities in Pinus Tabulaeformis]

An open-top chamber experiment was conducted to study the effects of high concentration O3 (80 nmol x mol(-1)) on the superoxide anion radical (O2*-) generation rate, hydrogen peroxide (H2O2) content, activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR) and glutathione reductase (GR), and ascorbic acid content in Pinus tabulaeformis leaves. Under high concentration O3 exposure, the superoxide anion radical generation rate and H2O2 and malondialdehyde (MDA) contents increased, while the ascorbic acid content and the activities of SOD, APX, DHAR, MDHAR, and GR increased in early growth season but decreased then to a level lower than the control, which illustrated that the antioxidant system of P. tabulaeformis did respond in an acclimation way in the early growth season, but could not bear the damage of long-term elevated O3 exposure.

Antagonistic HLH/bHLH Transcription Factors Mediate Brassinosteroid Regulation of Cell Elongation and Plant Development in Rice and Arabidopsis

In rice (Oryza sativa), brassinosteroids (BRs) induce cell elongation at the adaxial side of the lamina joint to promote leaf bending. We identified a rice mutant (ili1-D) showing an increased lamina inclination phenotype similar to that caused by BR treatment. The ili1-D mutant overexpresses an HLH protein homologous to Arabidopsis thaliana Paclobutrazol Resistance1 (PRE1) and the human Inhibitor of DNA binding proteins. Overexpression and RNA interference suppression of ILI1 increase and reduce, respectively, rice laminar inclination, confirming a positive role of ILI1 in leaf bending. ILI1 and PRE1 interact with basic helix-loop-helix (bHLH) protein IBH1 (ILI1 binding bHLH), whose overexpression causes erect leaf in rice and dwarfism in Arabidopsis. Overexpression of ILI1 or PRE1 increases cell elongation and suppresses dwarf phenotypes caused by overexpression of IBH1 in Arabidopsis. Thus, ILI1 and PRE1 may inactivate inhibitory bHLH transcription factors through heterodimerization. BR increases the RNA levels of ILI1 and PRE1 but represses IBH1 through the transcription factor BZR1. The spatial and temporal expression patterns support roles of ILI1 in laminar joint bending and PRE1/At IBH1 in the transition from growth of young organs to growth arrest. These results demonstrate a conserved mechanism of BR regulation of plant development through a pair of antagonizing HLH/bHLH transcription factors that act downstream of BZR1 in Arabidopsis and rice.

Comparison of Methods for Correcting Population Stratification in a Genome-wide Association Study of Rheumatoid Arthritis: Principal-component Analysis Versus Multidimensional Scaling

ABSTRACT : Population stratification (PS) represents a major challenge in genome-wide association studies. Using the Genetic Analysis Workshop 16 Problem 1 data, which include samples of rheumatoid arthritis patients and healthy controls, we compared two methods that can be used to evaluate population structure and correct PS in genome-wide association studies: the principal-component analysis method and the multidimensional-scaling method. While both methods identified similar population structures in this dataset, principal-component analysis performed slightly better than the multidimensional-scaling method in correcting for PS in genome-wide association analysis of this dataset.

Genome-wide Association Study for Empirically Derived Metabolic Phenotypes in the Framingham Heart Study Offspring Cohort

ABSTRACT : We used data reduction and clustering methods to identify five phenotypically homogeneous groups of study participants with similar profiles for cardiovascular disease risk factors. We constructed both qualitative (binary subgroup membership) and quantitative traits (probability of subgroup membership) for each individual. The Cluster 1 comprised individuals who were generally healthy and had some history of smoking. Cluster 2 was dropped from the analyses due to the preponderance of missing data. Cluster 3 was used as the control group, healthy non-smokers. Members of Cluster 4 had features of the metabolic syndrome and were generally not as obese as Cluster 5. Obesity was the hallmark of Cluster 5, the members of which also had some features of the metabolic syndrome.We then examined the genetic associations with both qualitative and quantitative representations of these empirically derived traits. Genetic analyses of the qualitative traits were conducted, comparing each of the affected groups with the unaffected cluster alone and, to increase statistical power, the unaffected group and healthy smokers combined. One single-nucleotide polymorphism on chromosome 4 met a conservative genome-wide significance level, but the effect was muted when we accounted for population stratification. The results for the quantitative traits were similar, with a small number of genome-wide significant findings muted by control for admixture. The directional findings will provide the basis for hypothesis generation for syndromes such as the metabolic syndrome and obesity.

[Role of Transforming Growth Factor Beta1 in the Development of Atrophic Gastritis]

To investigate the role of transforming growth factor beta1 (TGFbeta1) in the development of Helicobacter pylori (H.pylori)-associated non-metaplastic atrophic gastritis.

Antibody-drug Conjugates As Targeted Cancer Therapeutics

Traditional chemotherapy has become one of the essential treatments of cancer. However, cytotoxic agents are not tumor specific, which would cause serious side effects. Antibody-drug conjugates (ADCs), also called immunoconjugates, belong to the "targeted chemotherapeutics" category of anti-cancer drugs. ADCs are composed of three components including the cytotoxic drug, the monoclonal antibody, and the linker connecting the drug to the antibody. With the special-binding between antibody and antigen expressed on the surface of targeted cancer cells, ADCs provide a method to achieve excellent localization of the drug at the desired site in the body. The internalization and formation of ADCs are crucial in designing and applying an antibody conjugate to a particular disease model. In this review, we summarize three distinct internalization routes of ADCs and analysis the structure of ADCs. We also discuss in detail the categories and interaction of every component, as well as their influence to targeting property, liability and activity.

Defective Phagocytosis of Apoptotic Cells by Macrophages in Atherosclerotic Lesions of Ob/ob Mice and Reversal by a Fish Oil Diet

The complications of atherosclerosis are a major cause of death and disability in type 2 diabetes. Defective clearance of apoptotic cells by macrophages (efferocytosis) is thought to lead to increased necrotic core formation and inflammation in atherosclerotic lesions.

Isoflurane Preconditioning Ameliorates Endotoxin-induced Acute Lung Injury and Mortality in Rats

The effects of isoflurane pretreatment on pulmonary proinflammatory cytokines and survival in severe endotoxin-induced acute lung injury (ALI) have not been studied systemically. We investigated the effect of preadministration of isoflurane on ALI induced by lipopolysaccharide (LPS) in rats.

[The Expression of the Capsid Protein VP2 from Human Bocavirus Identified in Beijing and the Formation of Virus-like Particles (VLPs) in Insect Cells]

The aim of this study was to obtain the capsid protein VP2 of human bocavirus (HBoV) identified in Beijing recently and construct virus-like particles (VLPs) in insect cells for further study of this virus. The full-length VP2 gene of HBoV from BJ3722 was inserted into the baculovirus expression transfer vector (pFastBac 1) to obtain the recombinant Bacmid, and generation of recombinant baculoviruses was followed by transfection of the recombinant Bacmid into insect cells. Then the recombinant VP2 protein was recognized by SDS-PAGE using Coomassie-blue staining and Western blot using hyper-immune serum against VP2 of HBoV from rabbit. The recombinant baculoviruses were harvested and amplified to gain large amounts of viruses with high titers to infect insect cells at a multiplicity of infection (MOI) of 0. 5. After 7-10 days or 4-5 days of the infection, the supernatants of culture or the cell lysates treated with lysing solution were harvested, and ultracentrifuged twice through 40% sucrose cushion to obtain purified VLPs, which were followed by Western blot and IFA for VLPs' composition and specificity analysis, by electron microscopy for VLPs' morphologic structure. The recombinant VP2 protein with molecular weight of approximately 61 kD expressed in recombinant baculoviruses was recognized by SDS-PAGE using Coomassie-blue staining and Western blot. The presence of VP2 on VLPs was demonstrated by Western blot and IFA from samples collected during the purification of VLPs from the supernatants of culture or the cell lysates, and the expression of VP2 in insect cells led to the formation of VLPs which formed the typical icosahedral appearance of parvoviruses with a diameter of approximately 20 nm. In conclusion, the recombinant baculoviruses were constructed, the HBoV VP2 protein was expressed in insect cells with high specific antigenicity and VLPs was formed successfully.

[Env Gene Sequencing and HIV-1 Subtyping of Infected MSM from Beijing]

OBJECTIVE: To comprehend the latest HIV-I epidemic tendency and the character of V3 loop in MSM population of Beijing. METHODS; The C2-V3 regions of the HIV envelop gene were amplified by nest-PCR and sequenced from 11 HIV-l-infected MSM in Beijing in 2007. The subtype and sequences of V3 loop was analyzed. RESULTS There are 4 subtype B strains, 5 CRF AE, 1 CRFO7BC and 1 CRF15-01B strains within all 11 strains. There are five types of central motifs of the 11 samples, in which GPGR and GPGQ are most common. CONCLUSION: Recombination subtype of HIV-1 are spread extensively in MSM population of Beijing.

Inflammatory Stimulation and Hypoxia Cooperatively Activate HIF-1{alpha} in Bronchial Epithelial Cells: Involvement of PI3K and NF-{kappa}B

The transcription factor hypoxia-inducible factor (HIF)-1 plays a central physiological role in oxygen and energy homeostasis, and is activated during hypoxia by stabilization of the subunit HIF-1α. Recent studies have demonstrated that non-hypoxic stimuli can also activate HIF-1α in a cell-specific manner. Here, we demonstrate that stimulation of BEAS-2B cells and primary human bronchial epithelial cells by proinflammatory cytokines TNFα/IL-4 strongly induced expression and transcriptional activity of HIF-1α under normoxic conditions and amplified hypoxic HIF-1α activation. TNFα/IL-4 stimulated de novo HIF-1α gene transcription and translation rather than affected HIF-1α protein degradation and mRNA decay process. The activation of HIF-1α by TNFα/IL-4 was countered by the phosphoinositol 3-kinase (PI3K) inhibitor LY-294002 and rapamycin, an antagonist of mammalian target of rapamycin (mTOR), but not by inhibition of the MAPK pathway. In line, TNFα/IL-4 also activated NF-κB, whereas blocking of NF-κB by an inhibitor or silencing NF-κB subunit p65 attenuated HIF-1α activation by TNFα/IL-4. We also found the collaborative induction of VEGF, a potent angiogenic factor required for airway remodeling, by TNFα/IL-4 and hypoxia partially via HIF-1α pathway in BEAS-2B cells. This study reports the previously unsuspected collaborative regulation of HIF-1α by TNFα/IL-4 and hypoxia in bronchial epithelial cells partially via PI3K-mTOR and NF-κB pathway, and thereby will lead to the elucidation of the importance of HIF-1 in integrating inflammatory and hypoxic response in the pathogenesis of airway diseases.

Incidence of Creutzfeldt-Jakob Disease in Taiwan: a Prospective 10-year Surveillance

This study was performed to estimate the incidence of Creutzfeldt-Jakob Disease (CJD) in Taiwan from 1998 to 2007. Suspected cases of CJD were reported to the Taiwan Creutzfeldt-Jakob Disease Surveillance Unit, a nationwide, hospital-based case report system initiated since 1996 to prospectively conduct a CJD epidemiological study. Consecutive patients who met the diagnostic criteria recommended by the World Health Organization were enrolled. The clinical information of each suspected case was collected and case ascertainment was performed by an expert committee. A total of 123 sporadic CJD were identified without any iatrogenic or new variant CJD cases. The overall annual incidence rate (95% CI) was 0.55 (0.46-0.65) cases per million person-year. There was no statistically significant difference between the calendar year of disease onset (P = 0.97). The incidence rates were not significantly different between women and men (P = 0.63). Age was the main factor for the risk of CJD (P < 0.0001). Age-specific incidence rate increased after the age of 40 years with the peak being in the 70-79 years age group. Our data showed low annual incidence rate and high frequency of methionine homozygous prion protein genotype of sCJD in Taiwan. This report provided important epidemiological data on ethnic Chinese.

Cloning, Expression and Characterization of Xylose Isomerase, XylA, from Caldanaerobacter Subterraneus Subsp. Yonseiensis

The xylA gene, coding for xylose isomerase, from the extreme thermophile, Caldanaerobacter subterraneus subsp. yonseiensis was cloned, sequenced, and expressed in Escherichia coli. The nucleotide sequence of the xylA gene encoded a polypeptide of 438 residues with a calculated molecular weight of 50,170 Da. The purified XylA showed high sequence homology (92% identity) with that of Thermoanaerobacter thermohydrosulfuricus. The recombinant enzyme expressed in Escherichia coli was purified by heat treatment and gel chromatography. The purified enzyme was thermostable with optimal activity at 95 degrees C. The enzyme required divalent cations including Zn(2+) for its maximal activity and thermostability.

Detection of Multiple SNPs in Numerous Samples with Polyacrylamide Gel-based Microarray

Previously we have developed polyacrylamide gel-based DNA microarray to genotype single nucleotide polymorphism (SNP) in a large number of samples, which has been proved as a simple and robust platform for high-throughput SNP screening. Here we improved this method to detect multiple SNPs by introducing multiplex polymerase chain reaction (multiplex-PCR) and immobilizing the products of multiplex-PCR to fabricate gel-based microarray and applying universal dual-color detectors in hybridization. In this report, five SNPs (rs191296, rs2280073, rs17599165, rs17599416 and rs7660336) of GABRA4 gene were chosen and successfully analyzed with the improved platform. Our experiment demonstrated that 3-dimentional polyacrylamide gel-based microarray of multiplex-PCR products make the platform for multiple SNPs genotyping becoming more labor-saving and time-saving. Appling the universal dual-color fluorescent detectors can reduce the cost over two-thirds for multiple SNPs analysis. It is concluded that the multiplex-PCR combined with the gel-based microarray hybridized with universal dual-color fluorescent detectors is efficient, rapid and simple for the detection of a single nucleotide mismatch, and may be very competitive in the efficiency, fidelity and cost for constructing DNA microarrays.

[The Genetic Polymorphisms of 6 Short Tandem Repeat Loci in Native Han Population in Jiangxi Province]

To investigate the genetic polymorphisms of 6 short tandem repeats (STR) loci, namely, D6S1043, D2S1772, D7S3048, D22-GATA198B05, D8S1132 and D11S2368 in native Han population of Jiangxi province, China.

Inflammatory Cytokine Gene Polymorphisms Increase the Risk of Atrophic Gastritis and Intestinal Metaplasia

To investigate the effects of interleukin-8 (IL-8), macrophage migration inhibitory factor (MIF) gene polymorphisms, Helicobacter pylori (H. pylori) infection, on the risk of developing severe chronic atrophic gastritis (SCAG) and intestinal metaplasia (IM).

Lack of Association Between Total Serum Homocysteine and Extracranial Cerebral Flow

High homocysteine (Hcy) concentration is associated with slow coronary flow. This study examined the association between Hcy and hemodynamic status in the extracranial cerebral arteries in healthy individuals.

A Blue-emitting Organic Compound 9-hydroxyl-3-hydroxyethyl-2-methyl-4H-pyrido[1,2-a]pyrimidin-4-one: Synthesis, Crystal Structure and Luminescent Properties

A blue-emitting organic compound, 9-hydroxyl-3-hydroxyethyl-2-methyl-4H-pyrido[1,2-a]pyrimidin-4-one (I), was synthesized by the reaction of 2-amino-3-hydroxylpridine with 2-acetylbutyrolactone. The crystal of the title compound in larger size was reported for the first time through the slow evaporation method. The whole molecule in the crystal is non-planar, but the two conjugated rings are almost co-planar (with an average dihedral angle of only about 1.621degrees). The hydroxyl group is in trans position to the pyrido[1,2-a]pyrimidin heterocyclic moiety. The molecules are linked by the N-HO, O-HO and C-HO hydrogen bonds into two-dimensional sheets. Additionally, the pi-pi interactions (average distance 3.3680 A) interconnect the sheets stabilizing the crystal structure. At room temperature, the compound exhibits an intense blue emission at 432 nm upon 323 nm excitation in the solid state. The simple EL device with the configuration of ITO/I + PMMA/Ag was fabricated, where the compound (I) was used as a main emitting material. The EL device fabricated had a maximum brightness of 289 cd m-2. The thermal stability of the compound was also investigated by thermogravimetric analysis (TGA).

Airway Management in a Neonate with Oral Teratoma

Age-related Changes in the Central Auditory System: Comparison of D-galactose-induced Aging Rats and Naturally Aging Rats

One of the most common complaints among aging individuals is difficulty in understanding speech in a compromised listening environment, such as when background noise is present. Age-related hearing loss (presbycusis) is associated with both peripheral and central neural processing deficits, as it occurs even in those with only a mild peripheral hearing impairment. The current study was designed to investigate potential causative mechanisms of this impairment by using a rat model in which presbycusis is inducible by administration of D-galactose (D-gal). One group of these rats was injected subcutaneously with 150 mg D-gal daily for 8 weeks, while control animals received vehicle only. These groups were compared to naturally aged rats (24 months) that had received no other treatment. Central auditory function of the three groups was evaluated by measuring the auditory brainstem response (ABR) and middle latency response (MLR). A TaqMan real time PCR assay was used to quantify a 4834-bp deletion in the mitochondrial DNA (mtDNA) of the auditory cortex (AC), inferior colliculus (IC) and cochlear nucleus (CN). We assessed changes in lipid peroxidation levels and apoptosis rates, and examined pathological changes corresponding to D-gal-induced aging and natural aging. Both groups of aged rats exhibited delayed ABR latencies (III, IV, V), MLR Pa latency, and I-IV interpeak latency. Moreover, increased mtDNA 4834 bp deletion rates, lipid peroxidation levels, rates of neuronal apoptosis and neurodegenerative changes in the AC, IC and CN were similar among the D-gal induced and NA rats. However, the threshold of ABR in the D-gal group showed no significant change from the control group. These observations suggest that age-related central auditory dysfunction and its corresponding pathological changes are present in both naturally aging rats and the D-gal mimetic aging model. Oxidative stress, large-scale mtDNA 4834 bp deletion, and apoptosis are likely to be involved in the progressive weakening of the central auditory system associated with the aging process.

In Situ Mechanical Characterization of Mouse Oocytes Using a Cell Holding Device

This paper presents a cellular force measurement technique that allows for mechanical characterization of mouse oocytes during microinjection (i.e., in situ) without requiring a separate characterization process. The technique employs an elastic cell holding device and a sub-pixel computer vision tracking algorithm to resolve cellular forces in real time with a nanonewton force measurement resolution (2 nN at 30 Hz). Mechanical properties (i.e., stiffness) of both healthy and defective mouse oocytes are characterized. The experimental results suggest that the in situ obtained force-deformation data are useful for distinguishing healthy mouse oocytes from those with aging-induced cellular defects, promising an approach for oocyte quality assessment during microinjection. Biomembrane and cytoskeleton structures of the healthy and defective oocytes are also investigated in an attempt to correlate the measured subtle mechanical difference to cellular structure changes.

Terminal Osseous Dysplasia is Caused by a Single Recurrent Mutation in the FLNA Gene

Terminal osseous dysplasia (TOD) is an X-linked dominant male-lethal disease characterized by skeletal dysplasia of the limbs, pigmentary defects of the skin, and recurrent digital fibroma with onset in female infancy. After performing X-exome capture and sequencing, we identified a mutation at the last nucleotide of exon 31 of the FLNA gene as the most likely cause of the disease. The variant c.5217G>A was found in six unrelated cases (three families and three sporadic cases) and was not found in 400 control X chromosomes, pilot data from the 1000 Genomes Project, or the FLNA gene variant database. In the families, the variant segregated with the disease, and it was transmitted four times from a mildly affected mother to a more seriously affected daughter. We show that, because of nonrandom X chromosome inactivation, the mutant allele was not expressed in patient fibroblasts. RNA expression of the mutant allele was detected only in cultured fibroma cells obtained from 15-year-old surgically removed material. The variant activates a cryptic splice site, removing the last 48 nucleotides from exon 31. At the protein level, this results in a loss of 16 amino acids (p.Val1724_Thr1739del), predicted to remove a sequence at the surface of filamin repeat 15. Our data show that TOD is caused by this single recurrent mutation in the FLNA gene.

The Effect of Mitomycin C in Reducing Intraarticular Adhesion After Knee Surgery in Rabbits

Mitomycin C (MMC) is known to inhibit fibroblast proliferation through suppressing DNA dependent RNA synthesis. Based on this knowledge, we illustrated the effect of MMC on inhibiting fibroblast collagen synthesis and reducing intraarticular fibrous adhesion in a rabbit model. Forty-eight New Zealand white rabbits were randomly divided into four groups. Approximately 10 x 10 mm squares of the cortical bone was removed from both sides of the femoral condyle, and the underneath cancellous bone was exposed. MMC in various concentrations or saline were then applied to the decorticated areas. The intraarticular adhesions were evaluated after four weeks by macroscopic evaluation, histological evaluation and biochemical analysis of hydroxyproline content. The results demonstrated that MMC could suppress the formation of intraarticular fibrous adhesion in a dose-dependent manner. The intraarticular adhesion score, the hydroxyproline content and the fibroblast number in 0.1mg/ml MMC group were significantly less than those in 0.05 mg/ml MMC group, 0.01 mg/ml MMC group and control group. However, dense adhesions were found in 0.01 mg/ml MMC group and control group. These results indicated that topical application of 0.1mg/ml MMC could reduce intraarticular adhesion through inhibiting fibroblast proliferation in rabbits.

Phospho-sulindac (OXT-328), a Novel Sulindac Derivative, is Safe and Effective in Colon Cancer Prevention in Mice

Nonsteroidal anti-inflammatory drugs (NSAIDs) are effective cancer chemopreventive agents. However, chronic administration of NSAIDs is associated with significant side effects, mainly of the gastrointestinal tract. Given these limitations, we synthesized phospho-sulindac (P-S; OXT-328), a novel sulindac derivative.

[Human Metapneumoviruses Were Isolated from Infants and Children with Acute Respiratory Infections in Beijing]

To obtain isolated human metapneumovirus (HMPV) strains from clinical specimens collected from infants and children in Beijing and to promote the investigation on this important respiratory pathogen.

[Effect of Hydrogen-rich Saline on Blood Pressure and Antioxidant Ability of Lung Tissue in Scalded Rats Following Delayed Resuscitation]

To study the effect of hydrogen-rich saline on blood pressure and antioxidant ability of lung tissue in scalded rats following delayed resuscitation.

Quantitative in Vivo Brain Magnetic Resonance Spectroscopic Monitoring of Neurological Involvement in Mucopolysaccharidosis Type II (Hunter Syndrome)

Neurological involvement in X-linked mucopolysaccharidosis type II (Hunter syndrome) is indicative of more severe disease, but is not attenuated by current enzyme replacement therapy which does not significantly penetrate the blood-brain barrier. Magnetic resonance spectroscopy is an objective method of determining brain metabolites and has the potential to identify disease biomarkers with utility in evaluating current and novel therapies. MRS studies from seven patients with MPSII all receiving enzyme replacement therapy were compared with a large cohort of children with various neurocognitive disorders with normal MR imaging. All studies were completed on 1.5Tesla clinical MR scanners. Brain metabolite concentrations were determined from basal ganglia and parieto-occipital white matter using LCModel quantification. Serial trends in brain metabolites were analysed. Examination of mean spectra and quantitative metabolite concentrations demonstrated significantly decreased white matter N-acetylaspartate (a neuronal marker), total choline and glutamate, and elevated myo-inositol (glial marker) in MPSII patients. Analysis of serial determinations of white matter N-acetylaspartate demonstrated no change in two patients with stable MR imaging features but decreasing N-acetylaspartate in two patients more severely affected or deteriorating. These data demonstrate the utility of MRS to monitor serial alterations in brain metabolites including N-acetylaspartate which could be used as biomarkers of progressive neurological disease in MPSII. Integrated as an adjunct to MRI, such an approach could aid the evaluation of the efficacy of current ERT and also novel CNS-targeted therapies in MPSII.

A Comparison Between Simulated and Experimental Basis Sets for Assessing Short-TE in Vivo ¹H MRS Data at 1.5 T

A number of algorithms designed to determine metabolite concentrations from in vivo (1)H MRS require a collection of single metabolite spectra, known as a basis set, which can be obtained experimentally or by simulation. It has been assumed that basis sets can be used interchangeably, but no systematic study has investigated the effects of small variations in basis functions on the metabolite values obtained. The aim of this study was to compare the results of simulated with experimental basis sets when used to fit short-TE (1)H MRS data of variable quality at 1.5 T. Two hundred and twelve paediatric brain tumour spectra were included in the analysis, and each was analysed twice with LCModel™ using a simulated and experimental basis set. To determine the influence of data quality on quantification, each spectrum was assessed and 152 were classified as being of 'good' quality. Bland-Altman statistics were used to measure the agreement between the two basis sets for all available spectra and only 'good'-quality spectra. Monte-Carlo simulations were performed to investigate the influence of minor shifts in metabolite frequencies on metabolite concentration estimates. All metabolites showed good agreement between the two basis sets, and the average metabolite limits of agreement were approximately ±3.84 mM for all available data and ±0.99 mM for good-quality data. Errors obtained from the Monte-Carlo analysis were found to be more accurate than the Cramer-Rao lower bounds (CRLB) for 12 of 15 metabolites when metabolite frequency shifting was considered. For the majority of purposes, a level of agreement of ±0.99 mM between simulated and experimental basis sets is sufficiently small for them to be used interchangeably. Multiple analyses using slightly modified basis sets may be useful in estimating fitting errors, which are not predicted by CRLBs.

[Gradual Algorithm of Postconditioning Reduced Reperfusion Injury Through Mitochondrion Pathway in Rats]

The effects of various postconditioning algorithm on reperfusion injury and the role of mitochondrion pathway were investigated in a rat model of reperfusion/injury.

Single Cell Deposition and Patterning with a Robotic System

Integrating single-cell manipulation techniques in traditional and emerging biological culture systems is challenging. Microfabricated devices for single cell studies in particular often require cells to be spatially positioned at specific culture sites on the device surface. This paper presents a robotic micromanipulation system for pick-and-place positioning of single cells. By integrating computer vision and motion control algorithms, the system visually tracks a cell in real time and controls multiple positioning devices simultaneously to accurately pick up a single cell, transfer it to a desired substrate, and deposit it at a specified location. A traditional glass micropipette is used, and whole- and partial-cell aspiration techniques are investigated to manipulate single cells. Partially aspirating cells resulted in an operation speed of 15 seconds per cell and a 95% success rate. In contrast, the whole-cell aspiration method required 30 seconds per cell and achieved a success rate of 80%. The broad applicability of this robotic manipulation technique is demonstrated using multiple cell types on traditional substrates and on open-top microfabricated devices, without requiring modifications to device designs. Furthermore, we used this serial deposition process in conjunction with an established parallel cell manipulation technique to improve the efficiency of single cell capture from ∼80% to 100%. Using a robotic micromanipulation system to position single cells on a substrate is demonstrated as an effective stand-alone or bolstering technology for single-cell studies, eliminating some of the drawbacks associated with standard single-cell handling and manipulation techniques.

[Analysis of Clinical Manifestations of 159 Hospitalized Children Infected with 2009 Novel Influenza A (H1N1) Virus]

To analyze the clinical characteristics of hospitalized pediatric patients infected with 2009 H1N1 influenza.

Electrical Power Free, Low Dead Volume, Pressure-driven Pumping for Microfluidic Applications

This paper presents a simple-to-construct, low dead volume pump capable of generating a wide range of positive and negative pressures for microfluidic applications. The pump generates pressure or vacuum by changing the volume of air confined inside a syringe and is able to generate pressures between -95 and +300 kPa with a resolution as high as 1 Pa. Different from syringe pumps and electrokinetic pumping, which are capable of controlling flow rates only, our pump can be used to generate constant flow rates or constant pressures, which are required for certain applications such as the aspiration of biological cells for biophysical characterization. Compared to syringe pumps, the new pump has almost zero dead volume and does not exhibit pulsatile flows. Additionally, the system does not require electrical power and is cost effective (∼$100). To demonstrate the capabilities of the pump, we used it to aspirate osteoblasts (MC3T3-E1 cells) and to determine Young's modulus of the cells, to generate a concentration gradient, and to produce variable-sized droplets in microchannels using hydrodynamic focusing.

Palmitate-induced Inhibition of Insulin Gene Expression in Rat Islet β-cells Involves the Ceramide Transport Protein

Endogenous ceramide plays an important role in the palmitate (Palm) impairment of proinsulin gene expression in pancreatic islet β-cells. Changes in the liposoluble ceramide levels not only depend on metabolic enzymes but also on its transport to subcellular sites in response to Palm stimuli. In this study, we show that suppression of ceramide transport protein (CERT) mRNA with small interfering RNA contributed to intracellular ceramide accumulation in response to chronic Palm exposure and impairment of proinsulin gene expression, similar to the effect of inhibiting ceramide scavenging enzyme sphingomyelin synthase (SMS). High dose Palm treatment increased protein kinase D (PKD)-induced phosphorylation of CERT and its dysfunction. Intracellular accumulation of ceramide was associated with reduction of PDX-1 nuclear localization and MafA protein levels and stimulation of CCAAT/enhancer binding protein β (C/EBP β) expression. These conditions also corresponded with a reduction of PDX-1 and MafA and an increase of C/EBP β binding to the insulin promoter. Furthermore, down-regulation of C/EBP β could block ceramide impairment of proinsulin gene expression. The results reveal that Palm-mediated dysfunction of ceramide transport may contribute to intracellular ceramide accumulation and result in dysfunction of pancreatic beta cells by affecting binding of transcription factors to the insulin promoter.

Integration of Brassinosteroid Signal Transduction with the Transcription Network for Plant Growth Regulation in Arabidopsis

Brassinosteroids (BRs) regulate a wide range of developmental and physiological processes in plants through a receptor-kinase signaling pathway that controls the BZR transcription factors. Here, we use transcript profiling and chromatin-immunoprecipitation microarray (ChIP-chip) experiments to identify 953 BR-regulated BZR1 target (BRBT) genes. Functional studies of selected BRBTs further demonstrate roles in BR promotion of cell elongation. The BRBT genes reveal numerous molecular links between the BR-signaling pathway and downstream components involved in developmental and physiological processes. Furthermore, the results reveal extensive crosstalk between BR and other hormonal and light-signaling pathways at multiple levels. For example, BZR1 not only controls the expression of many signaling components of other hormonal and light pathways but also coregulates common target genes with light-signaling transcription factors. Our results provide a genomic map of steroid hormone actions in plants that reveals a regulatory network that integrates hormonal and light-signaling pathways for plant growth regulation.

Phospho-sulindac (OXT-922) Inhibits the Growth of Human Colon Cancer Cell Lines: a Redox/polyamine-dependent Effect

Non-steroidal anti-inflammatory drugs such as sulindac are promising chemoprevention agents against colon cancer, but their weak potency and side effects limit their use for both chemoprevention and chemotherapy. Here, we evaluated the effect of a new sulindac derivative, phospho-sulindac or OXT-922, on the growth of human cancer cell lines and its mechanism of action. OXT-922 inhibited the growth of human cancer cell lines originating from colon, pancreas and breast ~11- to 30-fold more potently than sulindac. This effect was mediated by a strong cytokinetic effect. Compared with control, OXT-922 inhibited cell proliferation by up to 67%, induced apoptosis 4.1-fold over control and blocked the G(1) to S cell cycle phase transition. OXT-922 suppressed the levels of cell cycle regulating proteins, including cyclins D(1) and D(3) and Cyclin-dependent kinases (CDK) 4 and 6. The levels of intracellular reactive oxygen species (ROS), especially those of mitochondrial O₂ⁱ⁻, were markedly elevated (5.5-fold) in response to OXT-922. ROS collapsed the mitochondrial membrane potential and triggered apoptosis, which was largely abrogated by antioxidants. OXT-922 suppressed nuclear factor-kappaB activation and downregulated thioredoxin-1 expression. It also suppressed the production of prostaglandin E(2) and decreased cyclooxygenase-1 expression. Similar to sulindac, OXT-922 enhanced spermidine/spermine N(1)-acetyltransferase activity, reduced the cellular polyamine content and synergized with difluoromethylornithine to inhibit cancer cell proliferation and induce apoptosis. Our results suggest that OXT-922 possesses promising anticancer properties and deserves further evaluation.

A New Approach Utilizing Real-time QPCR to Detect in Vitro Base Excision Repair

DNA lesions in mammalian cells may be induced by reactive oxygen species, alkylation, and ionizing radiation. This damage can then be repaired via the base excision repair (BER) pathway, which includes single strand break repair (SSBR). Thus, the BER (SSBR) pathway plays a critical role in maintaining genomic integrity, and may help us to better understand the mechanisms of aging, tumor formation, and degenerative diseases. AP site (apurinic/apyrimidinic site) or damaged base excision, nucleotide insertion and ligation are included in the BER (SSBR) pathway, which are facilitated by different enzymes at each step. Most previous in vitro BER studies have used modified radiolabeled (32)P oligonucleotide substrates. Which is a very conventional method for in vitro BER assay. However, the use of radioactive isotope material was limited in various laboratories which are unable to handle radioactive hazard. In this study, we developed a novel technique using real-time quantitative PCR (qPCR) to quantify BER activity in in vitro assays. Single strand breaks, DNA ligase activity, and glycosylase activity were detected to establish the feasibility and advantages of this qPCR technique for in vitro BER profiling.

Protective Roles of Alpha-lipoic Acid in Rat Model of Mitochondrial DNA4834bp Deletion in Inner Ear

The protective roles of alpha-lipoic acid in the rat model of mitochondrial DNA (mtDNA) 4834bp deletion in inner ear were investigated. Forty female Wistar rats at 4 weeks of age were divided into four groups: group A (D-galactose group, n=10), group B (D-galactose+alpha-lipoic acid group, n=10), group C (alpha-lipoic acid group, n=10), and group D (control group, n=10). Auditory brainstem response (ABR) was used to detect the hearing threshold. Colorimetry was used to analyze activity of superoxide dismutase (SOD) and concentration of malondialdehyde (MDA). The percentage of mtDNA4834bp deletion in inner ear was identified by real-time PCR. There was no significant difference in ABR threshold shift among all groups. The percentage of mtDNA4834bp deletion in group A was higher than that in other groups, but there was no significant difference in percentage of mtDNA4834bp deletion among groups B, C, and D. The activity of SOD in group A was lower than that in other groups. The concentration of MDA in group A was higher than that in other groups. It was concluded that there was no significant hearing loss when the percentage of mtDNA4834bp deletion was lower than 12.5%. Alpha-lipoic acid could prevent the reactive oxygen species (ROS)-induced mtDNA4834bp deletion in inner ear of rats.

[Correlation and Risk Factors Between Vascular Endothelial Dysfunction and Erectile Dysfunction]

To study the correlation between risk factors of vascular endothelial dysfunction and erectile dysfunction(ED).

Integration of Light- and Brassinosteroid-signaling Pathways by a GATA Transcription Factor in Arabidopsis

Light and brassinosteroid (BR) antagonistically regulate the developmental switch from etiolation in the dark to photomorphogenesis in the light in plants. Here, we identify GATA2 as a key transcriptional regulator that mediates the crosstalk between BR- and light-signaling pathways. Overexpression of GATA2 causes constitutive photomorphogenesis in the dark, whereas suppression of GATA2 reduces photomorphogenesis caused by light, BR deficiency, or the constitutive photomorphogenesis mutant cop1. Genome profiling and chromatin immunoprecipitation experiments show that GATA2 directly regulates genes that respond to both light and BR. BR represses GATA2 transcription through the BR-activated transcription factor BZR1, whereas light causes accumulation of GATA2 protein and feedback inhibition of GATA2 transcription. Dark-induced proteasomal degradation of GATA2 is dependent on the COP1 E3 ubiquitin ligase, and COP1 can ubiquitinate GATA2 in vitro. This study illustrates a molecular framework for antagonistic regulation of gene expression and seedling photomorphogenesis by BR and light.

[Genomic Characterization of WU Polyomavirus Identified from Pediatric Patients with Acute Respiratory Infections in Beijing, China]

To characterize the genomic sequence and arrangement of WU polyomavirus (WU virus) identified in clinical specimens collected from children with acute respiratory infections in Beijing, China, the sequences of capsid proteins VP1, VP2, and the large tumor antigen (LTAg), as well as the 5'-terminal sequence of WU virus, were amplified from the clinical specimen with ID number of BJF5276 which was determined as WU virus positive by PCR amplification. The PCR amplicons were sequenced, and genomic sequence analysis was performed by using the software DNAStar. In addition, VP2 coding-region sequences were amplified from other 21 clinical specimens identified as WU virus positive to investigate the gene diversity of WU virus. The genomic sequence of WU virus BJF5276 with accession number of HQ218321 in GenBank was 5,229 base pairs in length with 3 major coding domain sequences (CDS) sited on one strand coding for capsid proteins VP2, VP3 and VP1, and two CDS sited on the complementary strand coding for small tumor antigen (STAg) and LTAg; These 22 VP2 CDS sequences including 5 sequences submitted to GenBank were compared with 64 corresponding sequences downloaded from GenBank by MegAlign of DNAStar software, indicated that these sequences coming from children in Beijing shared high homology (over 98.8%) with those from GenBank. Phylogenetic analysis of these VP2 CDS by using Neighbor-joining (NJ) analyses with 2,000 bootstraps (Mega 4.0) showed that 20 sequences out of 22 belonged to clade Ia, and other 2 of them belonged to clade III, including 1 clustered in IIIa and 1 in a novel cluster proposed as IIIc. In conclusion, the genomic sequence of WU polyomavirus detected from clinical specimens from children in Beijing is closely related to other WU polyomaviruses in the feature of genomic coding region arrangement. Overall variation of VP2 CDS was very low, and there were different clades circulating in Beijing with a dominant clade Ia, which is different from dominated Ib circulating in other parts of the world reported previously, and a novel clade IIIc was proposed.

TIMI Myocardial Perfusion Frame Count: a New Method to Assess Myocardial Perfusion and Its Predictive Value for Short-term Prognosis

We sought to develop a new quantitative method to evaluate the degree of myocardial perfusion.

[Establishment of Orthotopic Lung Cancer Model Expressing Enhanced Green Fluorescent Protein]

In vivo molecular imaging with mouse model could continuously and in real-time monitor the changes of the tumor. The aim of this study is to establish stable enhanced green fluorescent protein (EGFP) expressing NCI-H460 cell lines and relevant mouse model via orthotopic transplantation, and to study the characteristic of this model and the quantitative detection method of the primary tumor and metastatic lesions.

Surface Water Quality and Its Control in a River with Intensive Human Impacts--a Case Study of the Xiangjiang River, China

Surface water quality and its natural and anthropogenic controls in the Xiangjiang River were investigated using multivariate statistical approaches and a comprehensive observation dataset collected from 2004 to 2008. Cluster analysis (CA) grouped the 15 different sampling stations into five clusters with similar hydrochemistry characteristics and pollution levels. Four principal components (PCs), nutrients, heavy metals, natural components, and organic components, were extracted from the entire dataset. Comparison of the different regional characteristics of these four PCs revealed a decreasing trend for heavy metals and an increasing trend for organic factor on an annual scale, and the seasonal trend was only observed for natural factor. We also conducted analysis of variance (ANOVA) in combination with principal component analysis (PCA) to quantify the relative contribution of spatial and temporal variations to each of the four PCs. The results revealed that 62% of the contributions from the spatial sites were responsible for variations in heavy metals, while 83% of the contributions from the sampling time were responsible for natural variations observed. However, no significant spatial or temporal contributions were found to be responsible for the nutrient and organic variations. Finally, some suggestions regarding water management were put forward based on the current status and future trends of surface water quality in the Xiangjiang River.

Adult SVZ Lineage Cells Home to and Leave the Vascular Niche Via Differential Responses to SDF1/CXCR4 Signaling

Neural progenitor cells (NPCs) in the adult subventricular zone (SVZ) are associated with ependymal and vasculature niches, which regulate stem cell self-renewal and differentiation. Activated Type B stem cells and their progeny, the transit-amplifying type C cells, which express EGFR, are most highly associated with vascular cells, indicating that this niche supports lineage progression. Here, we show that proliferative SVZ progenitor cells home to endothelial cells in a stromal-derived factor 1 (SDF1)- and CXC chemokine receptor 4 (CXCR4)-dependent manner. We show that SDF1 strongly upregulates EGFR and alpha6 integrin in activated type B and type C cells, enhancing their activated state and their ability to bind laminin in the vascular niche. SDF1 increases the motility of type A neuroblasts, which migrate from the SVZ toward the olfactory bulb. Thus, differential responses to SDF1 can regulate progenitor cell occupancy of and exit from the adult SVZ vascular niche.

Activation of Hypoxia-inducible Factor-1alpha Via Nuclear Factor-kappa B in Rats with Chronic Obstructive Pulmonary Disease

Accumulating data suggested that hypoxia inducible factor (HIF)-1alpha plays an important role in the evolution and propagation of the inflammatory process. To characterize the activation of HIF-1alpha in rats with chronic obstructive pulmonary disease (COPD) and examine the possible role of nuclear factor (NF)-kappaB in this process, rats were challenged by introtracheal instillation of lipopolysaccharide (LPS) and exposure to cigarette smoke. Pyrrolidine dithiocarbamate (PDTC) was administered via the oral route 1 h before LPS or cigarettes administration. Four weeks later, pulmonary function and histology were tested; bronchoalveolar lavage fluid (BALF) and arterial blood gases were assayed. Activation of pulmonary NF-kappaB was assessed by quantitative PCR, immunoblot analysis, and electrophoretic mobility shift assay, respectively. Results showed that LPS and smog induced the characteristics of COPD seen in human. PDTC alleviated the development of COPD and the levels of cytokines in BALF of PDTC+COPD group were significantly decreased compared with that of COPD group. The activation of pulmonary NF-kappaB was inhibited by PDTC and the accumulation of HIF-1alpha gene expression in the COPD group was attenuated by PDTC pretreatment. Furthermore, the mRNA levels of HIF-1alpha target genes heme oxygenase-1 (HO-1) and vascular endothelial growth factor (VEGF) were parallel to the attenuation of HIF-1alpha by PDTC. These findings indicated that the activation of HIF-1alpha pathway via NF-kappaB contributes to the development of COPD, and administration of NF-kappaB inhibitor may attenuate the development of COPD.

Influence of Aging on the Calbindin-D-28k Immunoreactive Positive Dopaminergic Neurons in the Substantia Nigra Pars Compacta of Rats

We studied the relationship between aging and the vulnerability of substantia nigra pars compacta (SNc) calbindin-D-28k immunoreactive positive (CB+) dopaminergic (DA) neurons. Immunohistochemistry and cell counting were used to determine the number of CB+ DA neuron in aged rats (24 mon) compared to adult rats (5 mon). Furthermore, the expression of CB mRNA and protein levels in SN was studied by semi-quantitative RT-PCR and Western blotting. An 11% loss of CB+ DA neurons was detected in both the rostral (8.9%) and caudal (1.7%) segments but not in the intermedial segment of SNc in aged rats compared to adult rats (P<0.05). No difference was detected in CB mRNA and protein levels between aged and adult rats (P>0.05). These data suggest that expression levels of CB mRNA and protein may increase in the existing SNc DA neurons, which may compensate for the partial age dependent loss of CB+ DA neurons in the SNc.

Identification of WU Polyomavirus from Pediatric Patients with Acute Respiratory Infections in Beijing, China

A novel polyomavirus (WU virus) has been identified in pediatric patients with acute respiratory tract infections (ARI), but its role as a respiratory pathogen has not yet been demonstrated. To investigate if WU virus is related to acute respiratory infections in infants and children in Beijing, specimens collected from 674 pediatric patients with ARI from April 2007 to May 2008 and from 202 children without ARI were used for this investigation. Common respiratory viruses were tested by virus isolation and/or antigen detection by indirect immunofluorescent assay followed by RT-PCR or PCR for other viruses associated with respiratory infections in specimens collected from patients with ARI before WU virus DNA was detected. WU virus DNA was detected by initial screening and secondary confirmation PCR for all specimens. The region encoding the VP2 gene of the virus was amplified from 17 WU-virus-positive clinical specimens, and sequence analysis was performed. Thirty-eight of 674 (5.6%) specimens from patients with ARI and 3 of 202 (1.5%) specimens from children without ARI yielded PCR products with the predicted molecular weight, using either screening or confirmation primer sets, indicating that these specimens were WU virus positive. However, more than 60% of the 38 WU-virus-positive specimens from patients with ARI were also positive for one or more respiratory viruses. The nucleotide and deduced amino acid sequences of the region encoding the VP2 gene from 17 Beijing WU viruses shared high homology (>98.5%) with sequences from GenBank and among themselves. The data indicated that WU virus in Beijing occurred 3.7 times more frequently in pediatric patients with ARI than in those without ARI (p < 0.05).

Exploring the Concept of Aromaticity on Complexes of a Fourfold Benzannulated Cyclopentadienyl Ligand

The ligand dibenzo[c,g]fluorenide (Dbf(-)), combines a cyclopentadienide ligand and 1,1'-binaphthyl fragment in one molecule. Preliminary investigations confirmed the special electronic situation in this 6pi-electron donor by way of a series of novel transition-metal complexes. Herein, the electron delocalization was investigated in detail by means of DFT calculations in combination with calculations on the nucleus-independent chemical shifts (NICS). These results clearly prove that the Dbf(-) ion gains the largest aromatic stabilization among all benzannulated fluorenides, and there are two almost olefinic C=C bonds in this structure. These bonds undergo cyclopropanation when treated with ZnEt(2) and ClCH(2)I in a modified Simmons-Smith reaction.

Benign Metastasizing Leiomyoma: Report of Two Cases and Literature Review

A Microfabricated Platform for High-throughput Unconfined Compression of Micropatterned Biomaterial Arrays

High-throughput screening techniques for cellular response are often unable to account for several factors present in the in vivo environment, many of which have been shown to modulate cellular response to the screened parameter. Culture in three-dimensional biomaterials and active mechanical stimulation are two such factors. In this work, we integrate these microenvironmental parameters into a versatile microfabricated device, capable of simultaneously applying a range of cyclic, compressive mechanical forces to cells encapsulated in an array of micropatterned biomaterials. The fabrication techniques developed here are broadly applicable to the integration of three-dimensional culture systems in complex multilayered polymeric microdevices. Compressive strains ranging from 6% to 26% were achieved simultaneously across the biomaterial array. As a first demonstration of this technology, nuclear and cellular deformation in response to applied compression was assessed in C3H10T1/2 mouse mesenchymal stem cells encapsulated within poly(ethylene glycol) hydrogels. Biomaterial, cellular, and nuclear deformations were non-linearly related. Parametric finite element simulations suggested that this phenomenon was due to the relative stiffness differences between the hydrogel matrix and that of the encapsulated cell and nucleus, and to strain stiffening of the matrix with increasing compression. This complex mechanical interaction between cells and biomaterials further emphasizes the need for high-throughput approaches to conduct mechanically active experiments in three-dimensional culture.

Effect of Carvedilol on Coronary Flow Reserve in Patients with Hypertensive Left-ventricular Hypertrophy

Patients with hypertensive left-ventricular hypertrophy (LVH) have lower coronary flow reserve (CFR). Whether carvedilol can improve CFR of patients with hypertensive LVH is unknown. We aimed to investigate the effects of carvedilol on CFR in patients with hypertensive LVH.

Factors Affecting Survival in Adult Patients with Massive Burns

To identify treatment-related factors associated with mortality in massively burned adult patients.

A Phase 2 Study of Irinotecan, Cisplatin, and Simvastatin for Untreated Extensive-disease Small Cell Lung Cancer

BACKGROUND:: The objective of this study was to investigate the efficacy of simvastatin in combination with irinotecan and cisplatin in chemotherapy-naive patients with extensive-disease small-cell lung cancer (ED-SCLC). METHODS:: In this phase 2 study, 61 patients received treatment with irinotecan (65 mg/m(2)) and cisplatin (30 mg/m(2)) on Days 1 and 8 every 3 weeks until either death or disease progression occurred. Patients also received oral simvastatin (40 mg daily) during the course of chemotherapy. The primary endpoint was 1-year survival. Secondary endpoints included the response rate (RR), progression-free survival (PFS), and toxicity. RESULTS:: The 1-year survival rate was 39.3%. The median overall survival (OS) was 11 months, and the median PFS was 6.1 months. Overall, the RR was 75%. The most common grade 3/4 toxicity was neutropenia (67%). Efficacy of the treatment was associated significantly with smoking status. Compared with never-smokers, ever-smokers had a better RR (40% vs 78%; P = .01), a longer PFS (2.5 months vs 6.4 months; P = .018), and had a trend toward an improved OS (9.0 months vs 11.2 months; P = .095). The effect of smoking on survival was apparent when ever-smokers were subdivided according to pack-years (PY) of smoking. Ever-smokers who had smoked >65 PY had a significantly longer OS compared with ever-smokers who had smoked ≤65 PY or never-smokers (20.6 months vs 10.6 months vs 9.0 months, respectively; log-rank P = 0.032). In multivariate analysis, PY >65 was predictive of longer survival (hazard ratio, 0.280; 95% confidence interval, 0.113-0.694). CONCLUSIONS:: The current results indicated that simvastatin in combination with irinotecan and cisplatin did not improve the survival of patients with ED-SCLC. Although the subgroup analysis by smoking status was exploratory, the addition of simvastatin to irinotecan and cisplatin may improve the outcome of heavy smokers with ED-SCLC. Cancer 2010. © 2010 American Cancer Society.

40 Gbits/s All-optical Clock Recovery for Degraded Signals Using an Amplified Feedback Laser

All-optical clock recovery for the return-to-zero modulation format is demonstrated experimentally at 40 Gbits/s by using an amplified feedback laser. A 40 GHz optical clock with a root-mean-square (rms) timing jitter of 130 fs and a carrier-to-noise ratio of 42 dB is obtained. Also, a 40 GHz optical clock with timing jitter of 137 fs is directly recovered from pseudo-non-return-to-zero signals degraded by polarization-mode dispersion (PMD). No preprocessing stage to enhance the clock tone is used. The rms timing jitter of the recovered clock is investigated for different values of input power and for varying amounts of waveform distortion due to PMD.

Application of Cervical Arthroplasty with Bryan Cervical Disc: Long-term X-ray and Magnetic Resonance Imaging Follow-up Results

Cervical disc arthroplasty is a new technique for treating degenerative cervical disease. Its goal is to avoid the degeneration of adjacent levels by preserving motion at the treated level. The aims of this study were to evaluate the radiologic outcomes of Bryan cervical disc replacement and the degenerative status of adjacent segments.

[Identification and Typing of Adenoviruses from Pediatric Patients with Acute Respiratory Infections in Beijing from 2003 to 2008]

Adenovirus (ADV) is one of the most common causes of acute respiratory infections in infants and children. The objective of this study was to investigate the prevalence of adenovirus infection among pediatric patients with acute respiratory infections in Beijing and the types of the adenoviruses circulating in Beijing on the molecular bases.

Microfabricated Arrays for High-throughput Screening of Cellular Response to Cyclic Substrate Deformation

Mechanical forces play an important role in regulating cellular function and have been shown to modulate cellular response to other factors in the cellular microenvironment. Presently, no technique exists to rapidly screen for the effects of a range of uniform mechanical forces on cellular function. In this work, we developed and characterized a novel microfabricated array capable of simultaneously applying cyclic equibiaxial substrate strains ranging in magnitude from 2 to 15% to small populations of adherent cells. The array is versatile, and capable of simultaneously generating a range of substrate strain fields and magnitudes. The design can be extended to combinatorially manipulate other mechanobiological culture parameters in the cellular microenvironment. As a first demonstration of this technology, the array was used to determine the effects of equibiaxial mechanical strain on activation of the canonical Wnt/beta-catenin signaling pathway in cardiac valve mesenchymal progenitor cells. This high-throughput approach to mechanobiological screening enabled the identification of a novel co-dependence between strain magnitude and duration of stimulation in controlling beta-catenin nuclear accumulation. More generally, this versatile platform has broad applicability in the fields of mechanobiology, tissue engineering and pathobiology.

Cardiovascular Responses and Airway Complications Following Awake Nasal Intubation with Blind Intubation Device and Fibreoptic Bronchoscope: a Randomized Controlled Study

The blind intubation device (BID) was introduced for awake nasotracheal intubation recently. The aim of this study was to compare the cardiovascular responses and associated airway complications of fibreoptic bronchoscope (FOB) with those of BID. The intubation attempts and intubation time were also compared.

[Predictive Value of Cervical Length by Transvaginal Sonography for Preterm Pregnancy During Mid- and Late-trimester of Pregnancy]

To study the value of cervical length (CL) by transvaginal sonography in the mid-trimester and late-trimester for the prediction of preterm delivery.

Influence of Aging on the Dopaminergic Neurons in the Substantia Nigra Pars Compacta of Rats

To study the relationship between aging and the vulnerability of substantia nigra pars compacta (SNc) tyrosine hydroxylase immunoreactive positive (TH+) dopaminergic (DA) neurons. We determined the number of TH(+) DA neurons in aged rats (24 mon) compared to adult rats (5 mon) using immunohistochemistry and cell counting. Furthermore, the expression of TH mRNA and protein levels in SN was studied by semi-quantitative RT-PCR and Western blotting. A 13.6% loss of neurons was detected in rostral segment of SNc, and the expression of TH mRNA levels was also reduced (P < 0.05), however, no difference was detected in TH protein levels (P > 0.05). These data suggest that expression of TH protein may increase in the existing SNc DA neurons, which may compensate for the partial loss of TH+ DA neurons.

Effects of Dihydroxylphenyl Lactic Acid on Inflammatory Responses in Spinal Cord Injury

The initial mechanical tissue disruption of spinal cord injury (SCI) is followed by a period of secondary injury that increases the size of the lesion. Secondary injuries are associated with edema, inflammation, excessive cytokine release, excitotoxicity and cell apoptosis. 3,4-dihydroxyphenyl lactic acid (DLA) is one of the major water-soluble components of chemical constituents from Salvia miltiorrhiza (SM). To investigate the inhibition effects of DLA on secondary injury of SCI, focusing especially on suppression of inflammatory responses and the mechanism of this effect, the following studies were performed: Basso, Beattie, and Bresnahan (BBB) scores to assess motor functions till 10 days after SCI; Nissl and Fast Blue histological staining and immunohistochemistry of inhibitory-kappa B-alpha (IκB-α) and nuclear factor-kappa B (NF-κB) p65 subunit protein; levels of myeloperoxidase (MPO) activity analysis as an indicator of polymorphonuclear infiltration; IL-6 production in plasma 10 days after SCI; Western blot analysis to determine cytoplasm levels of IκB-α and NF-κB p65 subunit proteins in the nuclear fractions 10 days after SCI. DLA significantly attenuated the motor function and tissue damage following SCI in rats, significant reduced polymorphonuclear cell infiltration and IL-6 production, as well as reduced cytoplasm IκB-α degradation and the nuclear translocation of NF-κB p65 subunit protein after SCI. In conclusion, the results clearly demonstrate that DLA inhibit the inflammation responses induced by SCI via inhibiting effect of production of IL-6 and nuclear translocation of NF-κB.

The "X-Factor" Index: a New Parameter for the Assessment of Adolescent Idiopathic Scoliosis Correction

The correction rate (CR) and fulcrum bending correction index (FBCI) based on the fulcrum bending radiograph (FBR) were parameters introduced to measure the curve correcting ability; however, such parameters do not account for contributions by various, potential extraneous "X-Factors" (e.g. surgical technique, type and power of the instrumentation, anesthetic technique, etc.) involved in curve correction. As such, the purpose of the following study was to propose the concept of the "X-Factor Index" (XFI) as a new parameter for the assessment of the correcting ability of adolescent idiopathic scoliosis (AIS). A historical cohort radiographic analysis of the FBR in the setting of hook systems in AIS patients (Luk et al. in Spine 23:2303-2307, 1998) was performed to illustrate the concept of XFI. Thirty-five patients with AIS of the thoracic spine undergoing surgical correction were involved in the analysis. Plain posteroanterior (PA) plain radiographs were utilized and Cobb angles were obtained for each patient. Pre- and postoperative PA angles on standing radiograph and preoperative fulcrum bending angles were obtained for each patient. The fulcrum flexibility, curve CR, and FBCI were determined for all patients. The difference between the preoperative fulcrum bending angle and postoperative PA angle was defined as Angle(XF), which accounted for the correction contributed by "X-Factors". The XFI, designed to measure the curve correcting ability, was calculated by dividing Angle(XF) by the fulcrum flexibility. The XFI was compared with the curve CR and FBCI by re-evaluating the original data in the original paper (Luk et al. in Spine 23:2303-2307, 1998). The mean standing PA and FBR alignments of the main thoracic curve were 58.3° and 24.5°, respectively. The mean fulcrum flexibility was 58.8%. The mean postoperative standing PA alignment was 24.7°. The mean curve CR was 58.0% and the mean FBCI was 101.1%. The mean XFI was noted as 1.03%. The CR was significantly positively correlated to curve flexibility (r = 0.66; p < 0.01).The FBCI (r = -0.47; p = 0.005) and the XFI (r = -0.45; p = 0.007) were significantly negatively correlated to curve flexibility. The CR was not correlated to Angle(XF) (r = 0.29; p = 0.089).The FBCI (r = 0.97; p < 0.01) and the XFI (r = 0.961; p < 0.01) were significantly positively correlated to Angle(XF). Variation in XFI was noted in some cases originally presenting with same FBCI values. The XFI attempts to quantify the curve correcting ability as contributed by "X-Factors" in the treatment of thoracic AIS. This index may be a valued added parameter to accompany the FBCI for comparing curve correction ability among different series of patients, instrumentation, and surgeons. It is recommended that the XFI should be used to document curve correction, compare between different techniques, and used to improve curve correction for the patient.

[National Examinations for Dentist in Japan and Implications]

Questions such as the law that was accorded by National Board Examinations for Dentist in Japan, the qualification of the examinee, the organization of the committee, the implementation of the exam, testing items development and the eligibility criteria were introduced in details in this article. The evolution of the number of qualified candidates and the issues for development needed to be explored were proposed.

Graphic Patterns of Cortical Functional Connectivity of Depressed Patients on the Basis of EEG Measurements

Considerable evidences have shown a decrease of neu-ronal activity in the left frontal lobe of depressed patients, but the underlying cortical network is still unclear. The present study intends to investigate the conscious-state brain network patterns in depressed patients compared with control individuals. Cortical functional connectivity is quantified by the partial directed coherence (PDC) analysis of multichannel EEG signals from 12 depressed patients and 12 healthy volunteers. The corresponding PDC matrices are first converted into unweighted graphs by applying a threshold to obtain the topographic property in-degree (K(in)). A significantly larger K(in) in the left hemisphere is identified in depressed patients, while a symmetric pattern is found in the control group. Another two topographic measures, i.e., clustering coefficients (C) and characteristic path length (L), are obtained from the original weighted PDC digraphs. Compared with control individuals, significantly smaller C and L are revealed in the depression group, indicating a random network-like architecture due to affective disorder. This study thereby provides further support for the presence of a hemispheric asymmetry syndrome in the depressed patients. More importantly, we present evidence that depression is characterized by a loss of optimal small-world network characteristics in conscious state.

Virtually Transparent Epidermal Imagery for Laparo-endoscopic Single-site Surgery

This paper presents a novel design, and prototype implementation, of a virtually transparent epidermal imagery (VTEI) system for laparo-endoscopic single-site (LESS) surgery. The system uses a network of multiple, micro wireless cameras and multiview mosaicing technique to obtain a panoramic view of the surgery area. This view provides visual feedback to surgeons with large viewing angles and areas of interest so that the surgeons can improve the safety of surgical procedures by being better aware of where the surgical instruments are relative to tissue and organs. The prototype VTEI system also projects the generated panoramic view on the abdomen area to create a transparent display effect that mimics equivalent, but higher risk, open-cavity surgeries.

[Genetic Variability of Matrix (M), Small Hydrophobic (SH) and Attachment (G) Proteins of Human Metapneumovirus Circulating in Children in Beijing from 2006 to 2010]

Human metapneumovirus (hMPV) is associated with acute respiratory tract infections (ARTI) in all age groups. However, there is limited information of genetic analysis of hMPV circulating in Beijing. To learn the characteristics of structural protein genes of human metapneumovirus circulating in children in Beijing, sequence analysis of matrix (M), small hydrophobic (SH) and attachment (G) proteins of hMPV from 2006 to 2010 was performed. Phylogenetic analysis of nucleotide sequences of 42 full length M genes, 49 SH gene and 55 G gene revealed that the hMPVs from pediatric patients were divided into sub-genotypes A2, B1 and B. There were highly conserved identities among M gene, with 7 conserved mutations of amino acids between A and B genotypes which were fairly conserved in the same genotype A or B. The amino acid identities of SH were 60.7% to 64.4% between different genotypes, 93.3% - 100% among same sub-genotype and 84.7% - 88.7% between different sub-genotypes. Use of alternative transcription-termination codon, nucleotide deletion and insertion resulted in variable length of nucleotide and deduced amino acid of G protein. Amino acid identities within same genotype ranged from 81.5% - 100%, whereas sequence identities between two genotypes ranged from 34.0% - 38.6% at the amino acid level. A new cluster of G genes in sub-genotype B2 appeared due to the same mutations and insertion of two amino acids in G protein encoding genes amplified from specimens collected from 2008 to 2010. Prediction of antigen sites of SH and G protein indicated that the variation of antigen sites between different sub-genotypes existed.

Controlled Synthesis and Up-conversion Emission of Rare-earth Tri-doped NaYF4 Nanocrystals Under Femtosecond-laser Excitation

Cubic nanocrystal and hexagonal micro-rods NaYF4, with predictable size, shape and phase, have been successfully synthesized through hydrothermal reaction. The growth mechanism and the effect of mass transfer on the morphology of hexagonal micro-prism are both discussed in detail. The increase of tri-doping lanthanide ion concentration decreased the size of crystal particle, which was explained by the Arrhenius rate equation together combined with the Gibbs-Thomson relationship. Furthermore, the dopants did not only affect the sizes of tri-doped NaYF4 micro-rods, but also impacted upon fluorescence intensity. The fluorescence of tri-doped NaYF4: Nd3+/Yb3+/Er3+ system, excited by an 800 nm femtolaser, was intensified with the increase of doped lanthanide ions concentration. Nevertheless owing to the fluorescence quenching, the other two systems (NaYF4: Nd3+/Ho3+/Er3+ and NaYF4: Nd3+/Tm3+/Er3+) did not show the same phenomenon.

β-Pix Modulates Actin-mediated Recruitment of Synaptic Vesicles to Synapses

Presynaptic compartments are formed through the recruitment of preassembled clusters of proteins to points of cell-cell contact, however, the molecular mechanism(s) underlying this process remains unclear. We demonstrate that clusters of polymerized actin can recruit and maintain synaptic vesicles to discrete sites along the axon, and that cadherin/β-catenin/scribble/β-pix complexes play an important role in this event. Previous work has demonstrated that β-catenin and scribble are important for the clustering of vesicles at synapses. We demonstrate that β-pix, a Rac/Cdc42 guanine nucleotide exchange factor (GEF), forms a complex with cadherin, β-catenin, and scribble at synapses and enhances localized actin polymerization in rat hippocampal neurons. In cells expressing β-pix siRNA or dominant-negative β-pix that lacks its GEF activity, actin polymerization at synapses is dramatically reduced, and synaptic vesicle localization is disrupted. This β-pix phenotype can be rescued by cortactin overexpression, suggesting that β-pix-mediated actin polymerization at synapses regulates vesicle localization.

Prediction of Infarct Growth and Neurologic Deterioration in Patients with Positive Perfusion-diffusion Mismatch

BACKGROUND: To assess the value of baseline clinical severity and perfusion-diffusion mismatch as predictors for further infarct growth and clinical outcome. METHODS: Patients with acute ischemic stroke and initial perfusion-diffusion mismatch within 72h were enrolled. Baseline perfusion defects on time-to-peak (TTP) and cerebral blood volume (CBV) maps were measured. Infarct volume and stroke severity were assessed by diffusion-weighted image (DWI) and NIHSS, and were repeatedly assessed 7days later. The predictive value of baseline NIHSS and perfusion defects on further infarct growth and neurologic deterioration was determined. RESULTS: Fifty-two patients (mean age 68.3±12.8years, 42% women) were enrolled. CBV defects were significantly associated with infarct growth (CBV, p=0.02). Initial stroke severity, but not TTP and CBV mismatch (p=0.65 and 0.76, respectively), significantly inversely correlated with neurologic deterioration (p=0.001). CONCLUSIONS: In patients with mismatch, those with severe symptoms initially are more likely to have infarct growth, while those with minor symptoms tend to suffer from larger extent of neurologic deterioration within 1week. CBV is associated with further infarct growth but not clinical deterioration.

[Role of Perforin in Severe Preeclampsia]

To investigate the possible role of perforin (PFN) in the pathogenesis of severe preeclampsia.

Autoantibodies Against the Catalytic Domain of BRAF Are Not Specific Serum Markers for Rheumatoid Arthritis

Autoantibodies to the catalytic domain of v-raf murine sarcoma viral oncogene homologue B1 (BRAF) have been recently identified as a new family of autoantibodies involved in rheumatoid arthritis (RA). The objective of this study was to determine antibody responses to the catalytic domain of BRAF in RA and other autoimmune diseases. The association between RA-related clinical indices and these antibodies was also assessed.

Reflection Symmetry Integrated Image Segmentation

This paper presents a new symmetry integrated region-based image segmentation method. The method is developed to obtain improved image segmentation by exploiting image symmetry. It is realized by constructing a symmetry token that can be flexibly embedded into segmentation cues. Interesting points are initially extracted from an image by the SIFT operator and they are further refined for detecting the global bilateral symmetry. A symmetry affinity matrix is then computed using the symmetry axis and it is used explicitly as a constraint in a region growing algorithm in order to refine the symmetry of the segmented regions. A multi-objective genetic search finds the segmentation result with the highest performance for both segmentation and symmetry, which is close to the global optimum. The method has been investigated experimentally in challenging natural images and images containing man-made objects. It is shown that the proposed method outperforms current segmentation methods both with and without exploiting symmetry. A thorough experimental analysis indicates that symmetry plays an important role as a segmentation cue, in conjunction with other attributes like color and texture.

[Epidemiological Investigation of Burn Inpatients in 6 Hospitals of Hainan Province: a Retrospective Analysis in 8 Years]

To analyze epidemiological characteristics of burn inpatients in Hainan province over 8 years.

P53 Codon 72 Polymorphism and Colorectal Cancer: a Meta-analysis of Epidemiological Studies

Background/Aims: Studies investigating the association between p53 codon 72 polymorphism and colorectal cancer risk report conflicting results. In order to clarify this, we carried out a meta-analysis using published data to obtain more precise estimates of risk. Methodology: Electronic searches of PubMed and EMBASE were conducted to select studies for this meta- analysis. The principal outcome measure was the odds ratio (OR) with 95% confidence interval (CI) for the risk of colorectal cancer associated with p53 codon 72 genotype. Results: We identified seven epidemiological studies, which included 1964 colorectal cancer cases and 2943 controls. The combined results based on all studies showed that there was no significant difference in genotype distribution [Arg/Arg (OR=0.86, 95% CI=0.68, 1.08); Pro/Pro (OR=1.27, 95% CI=0.96, 1.68); Pro/Arg (OR=1.04, 95% CI=0.92, 1.17)] between colorectal cancer and non-cancer patients. When stratifying for race, we found that patients with colorectal cancer had a significantly higher frequency of Pro/ Pro (OR=1.79, 95% CI=1.37, 2.35) and lower frequency of Arg/Arg (OR=0.66, 95% CI=0.44, 0.98) than controls among Asians. No statistical association was found between this genotype and alcohol, tobacco, stage, histological differentiation, lymph node metastasis of colorectal cancer. Conclusions: Our meta-analysis suggests that the p53 codon 72 polymorphism may be associated with colorectal cancer among Asians.

A New RNA Vaccine Platform Based on MS2 Virus-like Particles Produced in Saccharomyces Cerevisiae

mRNA vaccines are potentially attractive alternatives to DNA vaccines more often discussed, as they are generally considered safer than their DNA counterparts. The major limitations on the potency of RNA vaccines are their instability and inability to spread in vivo. Virus-like particles (VLPs) based on the bacteriophage MS2 have demonstrated remarkably high stability and may provide an improved platform for RNA-based genetic vaccination. However, no in vivo study of an MS2 VLP-mediated RNA vaccine has been reported. Therefore, we developed a model vaccine wherein MS2 VLPs packaging HIV-1 gag mRNAs (1544 bases) were produced in Saccharomyces cerevisiae, and then, used to immunize BALB/c mice. Serological analyses showed that antigen-specific antibody responses were elicited by immunization. These findings suggest that MS2 VLPs can be used in the design and construction of novel and safe phage-based mRNA delivery vectors.

Phosphorylation State of Olig2 Regulates Proliferation of Neural Progenitors

The bHLH transcription factors that regulate early development of the central nervous system can generally be classified as either antineural or proneural. Initial expression of antineural factors prevents cell cycle exit and thereby expands the pool of neural progenitors. Subsequent (and typically transient) expression of proneural factors promotes cell cycle exit, subtype specification, and differentiation. Against this backdrop, the bHLH transcription factor Olig2 in the oligodendrocyte lineage is unorthodox, showing antineural functions in multipotent CNS progenitor cells but also sustained expression and proneural functions in the formation of oligodendrocytes. We show here that the proliferative function of Olig2 is controlled by developmentally regulated phosphorylation of a conserved triple serine motif within the amino-terminal domain. In the phosphorylated state, Olig2 maintains antineural (i.e., promitotic) functions that are reflected in human glioma cells and in a genetically defined murine model of primary glioma.

Phospho-ibuprofen (MDC-917) is a Novel Agent Against Colon Cancer: Efficacy, Metabolism, and Pharmacokinetics in Mouse Models

We have developed a novel chemical modification of conventional nonsteroidal anti-inflammatory drugs to reduce their toxicity and enhance their efficacy. Phospho-ibuprofen [(PI) 2-(4-isobutyl-phenyl)-propionic acid-4-(diethoxy-phosphoryloxy)-butyl ester (MDC-917)], a novel derivative of ibuprofen, strongly inhibited the growth of human colon cancer cells in vitro and SW480 human colon cancer xenografts in nude mice. PI was metabolized minimally by cultured cells, but extensively by liver microsomes and mice, undergoing regioselective oxidation to produce 1-OH-PI and carboxyl-PI, which can be hydrolyzed to 1-OH-ibuprofen and carboxyl-ibuprofen, respectively. PI also can be hydrolyzed to release ibuprofen, which can generate 2-OH-ibuprofen, carboxyl-ibuprofen, and ibuprofen glucuronide. After a single oral administration (400 mg/kg) of PI, ibuprofen and ibuprofen glucuronide are the main plasma metabolites of PI; they have, respectively, C(max) of 530 and 215 μM, T(max) of 1 and 2 h, elimination t(1/2) of 7.7 and 5.3 h, and area under the concentration-time curve (0-24 h) of 1816 and 832 μM × h. Intact PI was detected in several tissues but not in plasma; at a higher PI dose (1200 mg/kg), PI plasma levels were 12.4 μM. PI generated the same metabolites in mouse plasma as conventional ibuprofen, but with much lower levels, perhaps accounting for the enhanced safety of PI. The antitumor effect of PI was significantly associated with plasma ibuprofen levels (p = 0.016) but not with xenograft ibuprofen levels (p = 0.08), suggesting a complex anticancer effect. These results provide a pharmacological basis to explain, at least in part, the anticancer efficacy and safety of this promising compound and indicate that PI merits further evaluation as an anticancer agent.

High-spin Cyclopentadienyl Complexes: a Single-molecule Magnet Based on the Aryl-iron(II) Cyclopentadienyl Type

[Effects of Thymosin Alpha-1 on Radiation-induced Pneumonitis]

Radiation-induced lung injure is one of the major factors of limitation in radiotherapy for lung cancer. Whether the use of thymosin and radiotherapy simultaneously would increase the radiation-induced lung injure is unclear. The aim of this study is to evaluate the effects of thymosin alpha-1 on radiation induced pneumonitis in mice.

[Chest Radiographic Findings in Children with 2009 Influenza A (H1N1) Virus Infection]

To evaluate chest radiographic findings of children with 2009 influenza (H1N1) virus infection.

Hyperglycemia Induces Apoptosis of Pancreatic Islet Endothelial Cells Via Reactive Nitrogen Species-mediated Jun N-terminal Kinase Activation

Hyperglycemia significantly stimulates pancreatic islet endothelial cell apoptosis; however, the precise mechanisms are not fully understood. In the present study, treating pancreatic islet endothelial (MS-1) cells with high glucose (30mmol/l) but not mannitol significantly increased the number of apoptotic cells as compared with a physiological glucose concentration (5.5mmol/l). Hyperglycemia significantly stimulated the expression of inducible nitric oxide synthase (iNOS) and production of NO and peroxynitrite (ONOO(-)), relevant to MS-1 cell apoptosis. Moreover, induced reactive nitrogen species (RNS) significantly increased the expression of bax, cleaved caspase-3 and poly adenosine diphosphate (ADP)-ribose polymerase (PARP) via JNK activation, but the expression of bcl-2 was not altered. Furthermore, SP600125 (a specific inhibitor of JNK) and 1400W (a specific inhibitor of iNOS) significantly attenuated cell apoptosis induced by high glucose. Therefore, hyperglycemia triggers MS-1 cell apoptosis by activating an intrinsic-dependent apoptotic pathway via RNS-mediated JNK activation.

Engineered Tendon with Decellularized Xenotendon Slices and Bone Marrow Stromal Cells: an in Vivo Animal Study

The purpose of this study was to investigate an engineered composite of multilayer acellular tendon slices seeded with bone marrow stromal cells (BMSCs) as a possible solution for tendon reconstruction. BMSCs were harvested from 15 rabbits and infraspinatus tendons were harvested from 17 dogs. The decellularized tendons were sectioned in longitudinal slices with a thickness of 50 µm. The BMSCs were seeded on the slices and then the slices were bundled into one composite. The composite was implanted into a rabbit patellar tendon defect. Tendon slices without BMSCs were implanted into the contralateral patellar tendon as a control. The composites were evaluated by histology and qRT-PCR. The viability of BMSCs was assessed using a fluorescent marker. Histology showed viable cells between the collagen fibres on the cell-seeded side. Analysis by qRT-PCR showed higher tenomodulin, collagen type III, MMP3 and MMP13 expressions and lower collagen type I expression in the cell-seeded composite than in the tendon slices without BMSCs. We conclude that BMSCs can survive in a multilayer composite, express a tendon phenotype and enhance the metabolism of tendon in vivo. This in vivo study suggests a potential utility of this composite in tendon reconstruction. Copyright © 2011 John Wiley & Sons, Ltd.

Glucagon-like Peptide 1 Protects Microvascular Endothelial Cells by Inactivating the PARP-1/iNOS/NO Pathway

Increasing studies suggest that the activity of GLP-1 might be of significant importance in the development of type 2 diabetes beyond its serum glucose-lowering effects. However, to date, the anti-apoptosis mechanism by which GLP-1 acts on MILE SVEN 1 (MS-1) cells has not been fully explored with regard to the intracellular signaling pathway. Increasing evidence shows that apoptosis of islet microvascular endothelial cells (IMECs) participates in the pathogenesis of diabetes. We wondered whether GLP-1 exerts its anti-apoptosis effects by inactivating the PARP-1/iNOS/NO pathway in oxidized low-density-lipoprotein (oxLDL)-induced apoptosis in mouse IMECs (MS-1 cells), which may linked to GLP-1R/cAMP levels. MTT assay revealed that 2-h pre-incubation with GLP-1 markedly restored the oxLDL-induced loss of MS-1 viability in a concentration-dependent manner. This effect was accompanied by a significant decrease in intracellular nitric oxide (NO) activity. Moreover, GLP-1 suppressed lipid peroxidation, restored the activities of endogenous antioxidants, and decreased the level of NO. Pre-incubating MS-1 cells with GLP-1 reduced cell apoptosis. Finally, GLP-1 could efficiently prevent the upregulation of poly(ADP-ribose) polymerase-1/nitrotyrosine and inducible NO synthase protein. Simultaneously, the expression of GLP-1 receptor and the level of cAMP was consistent with the administration of GLP-1. Our findings suggest that GLP-1 can effectively protect MS-1 cells against oxLDL-induced apoptosis, which may be important in preventing the pathogenesis of diabetes mellitus.

Phospho-sulindac (OXT-328) Combined with Difluoromethylornithine Prevents Colon Cancer in Mice

The nonsteroidal anti-inflammatory drug (NSAID) sulindac and the ornithine decarboxylase (ODC) antagonist difluoromethylornithine (DFMO), individually and together, are effective inhibitors of colon carcinogenesis. However, chronic use of sulindac is associated with significant side effects. We evaluated the chemopreventive efficacy of phospho-sulindac (P-S, OXT-328), an apparently safe derivative of sulindac, together with DFMO, in HT-29 human colon cancer xenografts. Nude mice were divided into four groups as follows: group 1 received vehicle (corn oil); group 2 received P-S (100 mg/kg/d) by oral gavage; group 3 received DFMO (2% in drinking water); and group 4 received P-S (100 mg/kg/d) by gavage plus DFMO (2% in drinking water; P-S/DFMO). Eighteen days after implantation, compared with controls, tumor volume was inhibited 65.9% by P-S, 52.9% by DFMO, and 70.9% by P-S/DFMO (P < 0.01 for all). P-S/DFMO reduced cell proliferation 27.1% and increased apoptosis 38.9% compared with controls (P < 0.05 for both). Compared with controls, P-S reduced the levels of thioredoxin-1 (Trx-1) and thioredoxin reductase (TrxR), whereas DFMO reduced polyamine content (putrescine and spermidine) and TrxR levels. Importantly, P-S/DFMO decreased putrescine and spermidine levels and the expression of Trx-1, TrxR, and cyclooxygenase (COX) 2. Of these molecular targets, TrxR most consistently correlated with tumor growth. Study results show that P-S/DFMO is an efficacious drug combination for colon cancer prevention and also show the safety of P-S, which may overcome the limiting side effects of conventional sulindac. P-S/DFMO has an intricate mechanism of action extending beyond polyamines and including the thioredoxin system, an emerging regulator of chemoprevention. P-S/DFMO merits further evaluation.

Contribution of Common Deletion to Total Deletion Burden in Mitochondrial DNA from Inner Ear of D-galactose-induced Aging Rats

Mitochondrial DNA (mtDNA) mutations, especially deletions, have been suggested to play an important role in aging and degenerative diseases. In particular, the common deletion in humans and rats (4977bp and 4834bp deletion, respectively) has been shown to accumulate with age in post-mitotic tissues with high energetic demands. Among numerous deletions, the common deletion has been proposed to serve as a molecular marker for aging and play a critical role in presbyacusis. However, so far no previous publication has quantified the contribution of common deletion to the total burden of mtDNA deletions in tissues during aging process. In the present study, we established a rat model with various degrees of aging in inner ear induced by three different doses of d-galactose (d-gal) administration. Firstly, multiple mtDNA deletions in inner ear were detected by nested PCR and long range PCR. In addition to the common deletion, three novel mtDNA deletions were identified. All four deletions, located in the major arc of mtDNA, are flanked by direct repeats and involve the cytochrome c oxidase (COX) subunit III gene, encoded by mtDNA. Additionally, absolute quantitative real-time PCR assay was used to detect the level of common deletion and total deletion burden of mtDNA. The quantitative data show that the common deletion is the most frequent type of mtDNA deletions, exceeding 67.86% of the total deletion burden. Finally, increased mtDNA copy number, reduced COX activity and mosaic ultrastructural impairments in inner ear were identified in d-gal-induced aging rats. The increase of mtDNA replication may contribute to the accelerated accumulation of mtDNA deletions, which may result in impairment of mitochondrial function in inner ear. Taken together, these findings suggest that the common deletion may serve as an ideal molecular marker to assess the mtDNA damage in inner ear during aging.

Differential Genome-wide Profiling of Tandem 3' UTRs Among Human Breast Cancer and Normal Cells by High-throughput Sequencing

Tandem 3' UTRs produced by alternative polyadenylation (APA) play an important role in gene expression by impacting mRNA stability, translation, and translocation in cells. Several studies have investigated APA site switching in various physiological states; nevertheless, they only focused on either the genes with two known APA sites or several candidate genes. Here, we developed a strategy to study APA sites in a genome-wide fashion with second-generation sequencing technology which could not only identify new polyadenylation sites but also analyze the APA site switching of all genes, especially those with more than two APA sites. We used this strategy to explore the profiling of APA sites in two human breast cancer cell lines, MCF7 and MB231, and one cultured mammary epithelial cell line, MCF10A. More than half of the identified polyadenylation sites are not included in human poly(A) databases. While MCF7 showed shortening 3' UTRs, more genes in MB231 switched to distal poly(A) sites. Several gene ontology (GO) terms and pathways were enriched in the list of genes with switched APA sites, including cell cycle, apoptosis, and metabolism. These results suggest a more complex regulation of APA sites in cancer cells than previously thought. In short, our novel unbiased method can be a powerful approach to cost-effectively investigate the complex mechanism of 3' UTR switching in a genome-wide fashion among various physiological processes and diseases.

[A Hair Prosthesis Consisting of Allogeneic Hair and Polypropylene Mimicking Follicular Units: Long-term Result and Histocompatibility in Rabbits]

To develop a follicular unit-like construct with allogeneic hair, evaluate its histocompatibility and long-term stability after transplantation, and explore the possibility of its clinical application.

Robotic ICSI (intracytoplasmic Sperm Injection)

This paper is the first report of robotic intracytoplasmic sperm injection (ICSI). ICSI is a clinical procedure performed worldwide in fertility clinics, requiring pick-up of a single sperm and insertion of it into an oocyte (i.e., egg cell). Since its invention 20 years ago, ICSI has been conducted manually by a handful of highly skilled embryologists; however, success rates vary significantly among clinics due to poor reproducibility and inconsistency across operators. We leverage our work in robotic cell injection to realize robotic ICSI and aim ultimately, to standardize how clinical ICSI is performed. This paper presents some of the technical aspects of our robotic ICSI system, including a cell holding device, motion control, and computer vision algorithms. The system performs visual tracking of single sperm, robotic immobilization of sperm, aspiration of sperm with picoliter volume, and insertion of sperm into an oocyte with a high degree of reproducibility. The system requires minimal human involvement (requiring only a few computer mouse clicks), and is human operator skill independent. Using the hamster oocyte-human sperm model in preliminary trials, the robotic system demonstrated a high success rate of 90.0% and survival rate of 90.7% (n=120).

A Microfluidic Device for Simultaneous Electrical and Mechanical Measurements on Single Cells

This paper presents a microfluidic device for simultaneous mechanical and electrical characterization of single cells. The device performs two types of cellular characterization (impedance spectroscopy and micropipette aspiration) on a single chip to enable cell electrical and mechanical characterization. To investigate the performance of the device design, electrical and mechanical properties of MC-3T3 osteoblast cells were measured. Based on electrical models, membrane capacitance of MC-3T3 cells was determined to be 3.39±1.23 and 2.99±0.82 pF at the aspiration pressure of 50 and 100 Pa, respectively. Cytoplasm resistance values were 110.1±37.7 kΩ (50 Pa) and 145.2±44.3 kΩ (100 Pa). Aspiration length of cells was found to be 0.813±0.351 μm at 50 Pa and 1.771±0.623 μm at 100 Pa. Quantified Young's modulus values were 377±189 Pa at 50 Pa and 344±156 Pa at 100 Pa. Experimental results demonstrate the device's capability for characterizing both electrical and mechanical properties of single cells.

A Phase 2 Study of Irinotecan, Cisplatin, and Simvastatin for Untreated Extensive-disease Small Cell Lung Cancer

The objective of this study was to investigate the efficacy of simvastatin in combination with irinotecan and cisplatin in chemotherapy-naive patients with extensive-disease small-cell lung cancer (ED-SCLC).

[Prokaryotic Expression for Fusion Protein of Human Metapneumovirus and Its Preliminary Application As an Antigen for Antibody Detection]

To understand the effectiveness of prokaryotic expression of fusion protein (F) of human metapneumovirus (hMPV) and its application as antigen, F proteins from different genotypes of hMPV were expressed in prokaryotic expression system and purified by Ni-NTA affinity chromatography column. According to the hydrophobicity, antigen index and surface probability of F protein, the subunit 1 (F1) region of F protein was generated and expressed in E. Coil. BL21(DE3). The 6-His-F1 proteins with molecular weight of approximately 37 kD generated from hMPV of two genotypes were expressed efficiently mainly in inclusion body. The antigenicity and specificity of the expressed proteins were tested and confirmed by Western Blot using polyclonal antibody against hMPV and one serum specimen from a patient with confirmed hMPV acute infection,and polyclonal antibodies against human respiratory syncytial virus and parainfluenza virus 2 and 3. The results of preliminary use of the expressed proteins for detecting antibodies against hMPV in 457 serum specimens collected from different age groups in Beijing indicated that 66%-67% of sera in all age groups were positive. The positive rate of antibodies declined in children in age groups from birth to 2-year-old and then rose along with the increase in age, in which the lowest was in age group from 1 to 2-year-old and the highest in newborn and people older than 60 years. The data indicated the existence of maternal transferred antibodies against hMPV in infants and the risk of hMPV infections in children younger than 2 years old.

Prevalence of Human Herpesvirus 8 Infection in Systemic Lupus Erythematosus

For decades, scientists have tried to understand the environmental factors involved in the development of systemic lupus erythematosus (SLE), in which viral infections was included. Previous studies have identified Epstein-Barr virus (EBV) to incite SLE. Human herpesvirus 8 (HHV-8), another member of the gammaherpesvirus family, shares a lot in common with EBV. The characteristics of HHV-8 make it a well-suited candidate to trigger SLE.

MR Spectroscopy-based Brain Metabolite Profiling in Propionic Acidaemia: Metabolic Changes in the Basal Ganglia During Acute Decompensation and Effect of Liver Transplantation

Propionic acidaemia (PA) results from deficiency of Propionyl CoA carboxylase, the commonest form presenting in the neonatal period. Despite best current management, PA is associated with severe neurological sequelae, in particular movement disorders resulting from basal ganglia infarction, although the pathogenesis remains poorly understood. The role of liver transplantation remains controversial but may confer some neuro-protection. The present study utilises quantitative magnetic resonance spectroscopy (MRS) to investigate brain metabolite alterations in propionic acidaemia during metabolic stability and acute encephalopathic episodes.

Mitochondrial Transcription Factor A Overexpression and Base Excision Repair Deficiency in the Inner Ear of Rats with D-galactose-induced Aging

Oxidative damage to mtDNA is associated with excessive reactive oxygen species production. The mitochondrial common deletion (mtDNA 4977-bp and 4834-bp deletion in humans and rats, respectively) is the most typical and frequent form of mtDNA damage associated with aging and degenerative diseases. The accumulation of the mitochondrial common deletion has been proposed to play a crucial role in age-related hearing loss (presbycusis). However, the mechanisms underlying the formation and accumulation of mtDNA deletions are still obscure. In the present study, a rat mimetic aging model induced by D-Gal was used to explore the origin of deletion mutations and how mtDNA repair systems modulate this process in the inner ear during aging. We found that the mitochondrial common deletion was greatly increased and mitochondrial base excision repair capacity was significantly reduced in the inner ear in D-Gal-treated rats as compared with controls. The overexpression of mitochondrial transcription factor A induced by D-Gal significantly stimulated mtDNA replication, resulting in an increase in mtDNA copy number. In addition, an age-related loss of auditory sensory cells in the inner ear was observed in D-Gal-treated rats. Taken together, our data suggest that mitochondrial base excision repair capacity deficiency and an increase in mtDNA replication resulting from mitochondrial transcription factor A overexpression may contribute to the accumulation of mtDNA deletions in the inner ear during aging. This study also provides new insights into the development of presbycusis.

Apparent Diffusion Coefficient of Diffusion Weighted MRI in Endometrial Carcinoma-Relationship with Local Invasiveness

OBJECTIVE: To evaluate the relationship between apparent diffusion coefficient (ADC) value and the local invasiveness of endometrial carcinoma. METHODS AND MATERIALS: The MR imaging of seventy-three patients with endometrial carcinoma proved by post-operative pathology and sixty-four patients with healthy uteri were retrospectively reviewed. All MR examinations included axial T2WI and T1WI, sagittal T2WI and diffusion-weighted sequences (b=0 and b=1000s/mm(2)). Tumor size, mean ADC value (ADCm) and quartile ADC (ADCq) were acquired on post-processing workstation using voxel-analysis software. Differences between the ADC values among three layers of normal uterine body and endometrial carcinomas were compared by ANOVA test. Groups were divided according to pathologic type, histologic grade, depth of myometrial infiltration, presence of cervical invasion and lymphovascular space invasion, and lymph node metastasis. Tumor size and ADC values were compared and analyzed. RESULTS: ADC values were different in three zones of uterine body (P<0.001), with the lowest in junctional zone [(1.126±0.190)×10(-3)mm(2)/s] and highest in outer myometrium [(1.496±0.196)×10(-3)mm(2)/s]. Mean ADC value of endometrial carcinomas [(1.011±0.121)×10(-3)mm(2)/s] was lower than the normal uterine body. Quartile ADC and tumor size were greater in groups with more invasive pathologic factors (P<0.05). Deep myometrial infiltration, cervical invasion, lymphovascular space invasion and lymph node metastasis were more common as quartile ADC values and tumor sizes increased. CONCLUSION: Mean ADC value was lower in endometrial carcinoma was lower than the normal uterus. Quartile ADC, representing the intra-tumor heterogeneity of water movement, had a profound relationship with invasiveness of endometrial carcinomas, while mean ADC value did not. ADC values may serve as a quantitative indicator to complement routine sequences.

Two Polymorphs of (2-carboxyethyl)(phenyl)phosphinic Acid

Two polymorphs of (2-carboxyethyl)(phenyl)phosphinic acid, C(9)H(11)O(4)P, crystallize in the chiral P2(1)2(1)2(1) space group with similar unit-cell parameters. They feature an essentially similar hydrogen-bonding motif but differ slightly in their detailed geometric parameters. For both polymorphs, the unequivocal location of the hydroxy H atoms together with the expected differences in the P-O bond lengths establish unequivocally that both forms contain the S isomer; the protonated phosphinic acid and carboxy O atoms serve as hydrogen-bond donors, while the second phosphinic acid O atom acts as a double hydrogen-bond acceptor and the remaining carboxy O atom is not involved in hydrogen bonding. Thus, an undulating two-dimensional supramolecular layer aggregate is formed based on an R(4)(3)(20) ring unit. Such polymorphism derives from the rotation of the C-C single bonds between the two hydrogen-bond-involved carboxy and phosphinic acid moieties.

Oxidative Stress Mediates Through Apoptosis the Anticancer Effect of Phospho-nonsteroidal Anti-inflammatory Drugs: Implications for the Role of Oxidative Stress in the Action of Anticancer Agents

We assessed the relationship between oxidative stress, cytokinetic parameters, and tumor growth in response to novel phospho-nonsteroidal anti-inflammatory drugs (NSAIDs), agents with significant anticancer effects in preclinical models. Compared with controls, in SW480 colon and MCF-7 breast cancer cells, phospho-sulindac, phospho-aspirin, phospho-flurbiprofen, and phospho-ibuprofen (P-I) increased the levels of reactive oxygen and nitrogen species (RONS) and decreased GSH levels and thioredoxin reductase activity, whereas the conventional chemotherapeutic drugs (CCDs), 5-fluorouracil (5-FU), irinotecan, oxaliplatin, chlorambucil, paclitaxel, and vincristine, did not. In both cell lines, phospho-NSAIDs induced apoptosis and inhibited cell proliferation much more potently than CCDs. We then treated nude mice bearing SW480 xenografts with P-I or 5-FU that had an opposite effect on RONS in vitro. Compared with controls, P-I markedly suppressed xenograft growth, induced apoptosis in the xenografts (8.9 ± 2.7 versus 19.5 ± 3.0), inhibited cell proliferation (52.6 ± 5.58 versus 25.8 ± 7.71), and increased urinary F2-isoprostane levels (10.7 ± 3.3 versus 17.9 ± 2.2 ng/mg creatinine, a marker of oxidative stress); all differences were statistically significant. 5-FU's effects on tumor growth, apoptosis, proliferation, and F2-isoprostane were not statistically significant. F2-isoprostane levels correlated with the induction of apoptosis and the inhibition of cell growth. P-I induced oxidative stress only in the tumors, and its apoptotic effect was restricted to xenografts. Our data show that phospho-NSAIDs act against cancer through a mechanism distinct from that of various CCDs, underscore the critical role of oxidative stress in their effect, and indicate that pathways leading to oxidative stress may be useful targets for anticancer strategies.

Genetic Polymorphisms of a Novel Vascular Susceptibility Gene, Ninjurin2 (NINJ2), Are Associated with a Decreased Risk of Alzheimer's Disease

Accumulated evidences have shown that vascular risk factors, e.g., hypertension, diabetes mellitus and hyperlipidemia, may be related to the risk of dementia. This study investigated the association between genetic polymorphisms of a vascular susceptibility gene, Ninjurin2 (NINJ2), and the risk of dementia, which has not been explored previously.

Role of Claudins in Oxidant-induced Alveolar Epithelial Barrier Dysfunction

Claudins are the most important components of the tight junctions at the interface of the basolateral and apical membranes of polarized epithelial and endothelial cells. They determine the barrier properties of cell-cell contact existing between two neighboring cells and regulate paracellular permeability. Although maintenance of barrier properties requires intact epithelial tight junctions, relatively little is known about the role of claudins expressed in the alveolar epithelium in the regulation of epithelial permeability in response to inflammatory stimuli and oxidative stress, or injury. The present study was undertaken to determine whether differential expression of tight junction claudins is a mechanism for regulation of oxidant-induced pulmonary epithelial hyperpermeability. Here, we show that claudin-2 plays an important role in the regulation of epithelial barrier function during oxidative stress.

Characterization of Human Metapneumovirus from Pediatric Patients with Acute Respiratory Infections in a 4-year Period in Beijing, China

Human metapneumovirus (hMPV) was discovered by scientists in the Netherlands as a novel respiratory virus in 2001 and had been found in children with acute respiratory tract infections (ARTI) in China. The objective of this study was to determine the importance of hMPV infection in children in Beijing and the genotypes of the circulating virus by the surveillance during a four-consecutive-year period.

STK33 Kinase Activity is Nonessential in KRAS-dependent Cancer Cells

Despite the prevalence of KRAS mutations in human cancers, there remain no targeted therapies for treatment. The serine-threonine kinase STK33 has been proposed to be required for the survival of mutant KRAS-dependent cell lines, suggesting that small molecule kinase inhibitors of STK33 may be useful to treat KRAS-dependent tumors. In this study, we investigated the role of STK33 in mutant KRAS human cancer cells using RNA interference, dominant mutant overexpression, and small molecule inhibitors. As expected, KRAS downregulation decreased the survival of KRAS-dependent cells. In contrast, STK33 downregulation or dominant mutant overexpression had no effect on KRAS signaling or survival of these cells. Similarly, a synthetic lethal siRNA screen conducted in a broad panel of KRAS wild-type or mutant cells identified KRAS but not STK33 as essential for survival. We also obtained similar negative results using small molecule inhibitors of the STK33 kinase identified by high-throughput screening. Taken together, our findings refute earlier proposals that STK33 inhibition may be a useful therapeutic approach to target human KRAS mutant tumors.

E2F1 is Involved in DNA Single-strand Break Repair Through Cell-cycle-dependent Upregulation of XRCC1 Expression

The X-ray repair cross complementing group 1 (XRCC1) protein is involved in DNA base excision repair and its expression varies during the cell cycle. Although studies have demonstrated that rapid XRCC1-dependent single-strand break repair (SSBR) takes place specifically during S/G(2) phases, it remains unclear how it is regulated during the cell cycle. We found that XRCC1 is a direct regulatory target of E2F1 and further investigated the role of XRCC1 in DNA repair during the cell cycle. Saos2 primary osteosarcoma cells stably transfected with inducible E2F1-wt or mutant E2F1-132E were treated with hydroxurea (HU) for 36h and were subsequently withdrawn HU for 2-24h to test whether cell-cycle-dependent DNA SSBR requires E2F1-mediated upregulation of XRCC1. We found that SSBR activity, as determined using a qPCR-base method, was correlated with E2F1 levels at different phases of the cell cycle. XRCC1-positive (AA8) and negative (EM9) CHO cells were used to demonstrate that the alterations in SSBR were mediated by XRCC1. The results indicate that E2F1-mediated regulation of XRCC1 is required for cell-cycle-dependent SSBR predominantly in G(1)/S phases. Our observations have provided new mechanistic insight for understanding the role of E2F1 in the maintenance of genomic stability and cell survival during the cell cycle. The regulation of XRCC1 by E2F1 during cell-cycle-dependent SSBR might be an important aspect for practical consideration for resolving the problem of drug resistance in tumor chemotherapies.

[Sequence and Antigenicity Analysis for VP4s of EV71 Strains Isolated from Children in Beijing]

In order to learn about the correlation between the sequences of VP4 of EV71 and clinical symptoms of patients and analyze the antigenicity of VP4 of EV71, as well as the cross-reactivity with VP4 of CA16, the sequences of VP4 gene from 10 EV71 strains isolated from infants and children with hand, foot and mouth diseases (HFMD) during 2007 to 2009 were determined through standard molecular cloning protocols, and the results were analyzed by EditSeq and MegAlign of DNAStar. Full-length genes of VP4s of EV71 and CA16 were amplified from virus isolates and expressed in E. coli. Then the expressed VP4s were used as antigens to detect IgG antibody in 189 sera samples from people taking health check up and patients of non-HFMD by Western-Blot. They were also used to detect IgM antibody in 14 of sera samples from infants and children with EV71 infection and 12 of sera samples from those with CA16 infection. The nucleotides identities among these 10 sequences of VP4s isolated in our lab were 94.20% - 100.00% and the deduced amino acids were identical. There was no consistent divergence between the sequences of serious cases and those from general HFMD cases. Phylogenetic analysis based on VP4s indicated that these 10 VP4s of EV71 belonged to C4. The nucleotide identities between EV71 VP4 (s67) and CA16 VP4 (s401) was 69.60% and the deduced amino acids identities was 78.60%. In the detection of IgG, the sera-positive rate for EV71 VP4 was 38.10% and the sera-positive rate of CA16 VP4 was 58.20%. The difference in the sera-positive rate between them was significant (chi2 = 15.30, P < 0.01), suggesting that the expressed VP4s of EV71 and CA16 were of good antigenicity and not cross-reactive. There was no positive reaction detected for IgM against VP4s for EV71 or CA16. The data from this study reveal important information for the further study of EV71.

Automated Microinjection of Recombinant BCL-X into Mouse Zygotes Enhances Embryo Development

Progression of fertilized mammalian oocytes through cleavage, blastocyst formation and implantation depends on successful implementation of the developmental program, which becomes established during oogenesis. The identification of ooplasmic factors, which are responsible for successful embryo development, is thus crucial in designing possible molecular therapies for infertility intervention. However, systematic evaluation of molecular targets has been hampered by the lack of techniques for efficient delivery of molecules into embryos. We have developed an automated robotic microinjection system for delivering cell impermeable compounds into preimplantation embryos with a high post-injection survival rate. In this paper, we report the performance of the system on microinjection of mouse embryos. Furthermore, using this system we provide the first evidence that recombinant BCL-XL (recBCL-XL) protein is effective in preventing early embryo arrest imposed by suboptimal culture environment. We demonstrate that microinjection of recBCL-XL protein into early-stage embryos repairs mitochondrial bioenergetics, prevents reactive oxygen species (ROS) accumulation, and enhances preimplantation embryo development. This approach may lead to a possible treatment option for patients with repeated in vitro fertilization (IVF) failure due to poor embryo quality.

Motion-compensated Noncontact Imaging Photoplethysmography to Monitor Cardiorespiratory Status During Exercise

With the advance of computer and photonics technology, imaging photoplethysmography [(PPG), iPPG] can provide comfortable and comprehensive assessment over a wide range of anatomical locations. However, motion artifact is a major drawback in current iPPG systems, particularly in the context of clinical assessment. To overcome this issue, a new artifact-reduction method consisting of planar motion compensation and blind source separation is introduced in this study. The performance of the iPPG system was evaluated through the measurement of cardiac pulse in the hand from 12 subjects before and after 5 min of cycling exercise. Also, a 12-min continuous recording protocol consisting of repeated exercises was taken from a single volunteer. The physiological parameters (i.e., heart rate, respiration rate), derived from the images captured by the iPPG system, exhibit functional characteristics comparable to conventional contact PPG sensors. Continuous recordings from the iPPG system reveal that heart and respiration rates can be successfully tracked with the artifact reduction method even in high-intensity physical exercise situations. The outcome from this study thereby leads to a new avenue for noncontact sensing of vital signs and remote physiological assessment, with clear applications in triage and sports training.

Effect of Nanowire Number, Diameter, and Doping Density on Nano-FET Biosensor Sensitivity

Semiconductive nanowire-based biosensors are capable of label-free detection of biological molecules. Nano-FET (field-effect transistor) biosensors exhibiting high sensitivities toward proteins, nucleic acids, and viruses have been demonstrated. Rational device design methodologies, particularly those based on theoretical predictions, were reported. However, few experimental studies have investigated the effect of nanowire diameter, doping density, and number on nano-FET sensitivity. In this study, we devised a fabrication process based on parallel approaches and nanomanipulation-based post-processing for constructing nano-FET biosensor devices with carefully controlled nanowire parameters (diameter, doping density, and number). We experimentally reveal the effect of these nanowire parameters on nano-FET biosensor sensitivity. The experimental findings quantitatively demonstrate that device sensitivity decreases with increasing number of nanowires (4 and 7 nanowire devices exhibited a ∼38 and ∼82% decrease in sensitivity as compared to a single-nanowire device), larger nanowire diameters (sensors with 81-100 and 101-120 nm nanowire diameters exhibited a ∼16 and ∼37% decrease in sensitivity compared to devices with nanowire diameters of 60-80 nm), and higher nanowire doping densities (∼69% decrease in sensitivity due to an increase in nanowire doping density from 10(17) to 10(19) atoms·cm(-3)). These results provide insight into the importance of controlling nanowire properties for maximizing sensitivity and minimizing performance variation across devices when designing and manufacturing nano-FET biosensors.

Classification of Cell Types Using a Microfluidic Device for Mechanical and Electrical Measurement on Single Cells

This paper presents a microfluidic system for cell type classification using mechanical and electrical measurements on single cells. Cells are aspirated continuously through a constriction channel with cell elongations and impedance profiles measured simultaneously. The cell transit time through the constriction channel and the impedance amplitude ratio are quantified as cell's mechanical and electrical property indicators. The microfluidic device and measurement system were used to characterize osteoblasts (n=206) and osteocytes (n=217), revealing that osteoblasts, compared with osteocytes, have a larger cell elongation length (64.51 ± 14.98 μm vs. 39.78 ± 7.16 μm), a longer transit time (1.84 ± 1.48 s vs. 0.94 ± 1.07 s), and a higher impedance amplitude ratio (1.198 ± 0.071 vs. 1.099 ± 0.038). Pattern recognition using the neural network was applied to cell type classification, resulting in classification success rates of 69.8% (transit time alone), 85.3% (impedance amplitude ratio alone), and 93.7% (both transit time and impedance amplitude ratio as input to neural network) for osteoblasts and osteocytes. The system was also applied to test EMT6 (n=747) and EMT6/AR1.0 cells (n=770, EMT6 treated by doxorubicin) that have a comparable size distribution (cell elongation length: 51.47 ± 11.33 μm vs. 50.09 ± 9.70 μm). The effects of cell size on transit time and impedance amplitude ratio were investigated. Cell classification success rates were 51.3% (cell elongation alone), 57.5% (transit time alone), 59.6% (impedance amplitude ratio alone), and 70.2% (both transit time and impedance amplitude ratio). These preliminary results suggest that biomechanical and bioelectrical parameters, when used in combination, could provide a higher cell classification success rate than using electrical or mechanical parameter alone.

[Diagnosis and Treatment of Complicated Deep Infiltrated Endometriosis: a Case Report and Literature Review]

To explore diagnostic and therapeutic methods of patients with deep infiltrating endometriosis.

[Neuroendocrine Differentiation is Not a Malignant Index of Non-small Cell Lung Cancer]

A debate has been ongoing whether non-small cell lung cancer(NSCLC) with neuroendocrine(NE) differentiation likely indicates malignant behavior, poor prognosis, and sensitivity to chemotherapy. In response to this issue, we retrospectively investigated NE differentiation in NSCLC patients who underwent anatomical pulmonary surgery.

[Diagnosis and Treatment for Pulmonary Sclerosing Hemangioma]

The concept of pulmonary sclerosing hemangioma (PSH) was proposed only 50 years ago. PSH features several characteristics that should be differently diagnosed with lung cancer. The aim of this study is to retrospectively review the diagnosis and treatment of 48 cases of PSH, review recent publications about this topic, and address reasonable diagnosis and treatment methods.

A Polymeric Nanoparticle Encapsulated Hedgehog Pathway Inhibitor HPI-1 (NanoHHI) Inhibits Systemic Metastases in an Orthotopic Model of Human Hepatocellular Carcinoma

PURPOSE: To illustrate the prognostic significance of hedgehog (Hh) signaling in hepatocellular carcinoma (HCC) patients, and to evaluate the efficacy of a novel nanoparticle-encapsulated inhibitor of the Hh transcription factor, Gli-1 ("NanoHHI") using in vitro and in vivo models of human HCC.EXPERIMENTAL DESIGN: Patched1 (Ptch) expression was detected in tumor tissue microarrays of 396 HCC patients who underwent curative surgical resection during 2/2000 to 12/2002. Prognostic significance was assessed using Kaplan-Meier survival estimates and log-rank tests. The effects of NanoHHI alone and in combination with sorafenib were investigated on HCC cell lines. Primary HCC tumor growth and metastasis were examined in vivo using subcutaneous and orthotopic HCC xenografts in nude mice. RESULTS: Elevated expression of Ptch in HCC tissues was significantly related to disease recurrence, as well as a shorter time to recurrence in HCC patients. In vitro, NanoHHI significantly inhibited the proliferation and invasion of HCC cell lines. NanoHHI potently suppressed in vivo tumor growth of HCC xenografts in both subcutaneous and orthotopic milieus, and in contrast to sorafenib, resulted in significant attenuation of systemic metastases in the orthotopic setting. Further, NanoHHI significantly decreased the population of CD133-expressing HCC cells, which have been implicated in tumor initiation and metastases.CONCLUSIONS: Downstream Hh signaling has prognostic significance in HCC patients as it predicts early recurrence. Gli inhibition through NanoHHI has profound tumor growth inhibition and anti-metastatic effects in HCC models, which may provide a new strategy in the treatment of HCC patients and prevention post-operative recurrence.

Role of Caveolin-1 in the Regulation of Pulmonary Endothelial Permeability

Endothelial permeability measurements of intact vascular beds and monolayer cultures are used to describe transport of small molecules (ions, water, and nutrients), macromolecules and plasma protein across the vascular endothelia. Disruption of the endothelial barrier leads to vascular hyper-permeability and protein-rich edema which is a key hallmark of inflammation. Transport of the most abundant plasma protein, albumin, occurs by means of transcellular and paracellular pathways. In healthy, noninflamed vessels, endothelial cell-cell contacts significantly restrict the paracellular permeability of albumin, whereas its transcellular transport from the blood to the abluminal perivascular interstitium occurs via caveolae. Thus, caveolae-mediated transport is a primary determinant of the basal endothelial permeability properties. Increased paracellular permeability induced during inflammation is thought to be due to the opening of interendothelial cell-cell junctions and disruption of endothelial cell-matrix contacts within the vasculature. We recently demonstrated that caveolae-mediated transendothelial transport (transcytosis) of macromolecules through the microvascular endothelial barrier is also an important mechanism responsible for inflammation-evoked pulmonary vascular hyperpermeability and protein-rich edema formation. Moreover, caveolin-1, a structural and scaffolding protein required for caveolae formation and transcellular transport, also plays an important role in oxidant-induced paracellular hyperpermeability. This review highlights the methods used to assess transcellular and paracellular permeability properties of the intact mouse lung and cultured endothelial cell monolayers.

Nasotracheal Intubation Using the Blind Intubation Device in Anaesthetised Adults with Mallampati Class 3: a Comparison with the Macintosh Laryngoscope

We hypothesised that the Blind Intubation Device (BID) would be effective for nasotracheal intubation (NTI) in anaesthetised adults with Mallampati class 3. We also hypothesised that BID may cause less haemodynamic changes due to the avoidance of direct stimulation induced by the Macintosh blade.

(Micro)managing the Mechanical Microenvironment

Mechanical forces are critical components of the cellular microenvironment and play a pivotal role in driving cellular processes in vivo. Dissecting cellular responses to mechanical forces is challenging, as even "simple" mechanical stimulation in vitro can cause multiple interdependent changes in the cellular microenvironment. These stimuli include solid deformation, fluid flows, altered physical and chemical surface features, and a complex transfer of loads between the various interacting components of a biological culture system. The active mechanical and biochemical responses of cells to these stimuli in generating internal forces, reorganizing cellular structures, and initiating intracellular signals that specify cell fate and remodel the surrounding environment further complicates cellular response to mechanical forces. Moreover, cells present a non-linear response to combinations of mechanical forces, materials, chemicals, surface features, matrix properties and other effectors. Microtechnology-based approaches to these challenges can yield key insights into the mechanical nature of cellular behaviour, by decoupling stimulation parameters; enabling multimodal control over combinations of stimuli; and increasing experimental throughput to systematically probe cellular response. In this critical review, we briefly discuss the complexities inherent in the mechanical stimulation of cells; survey and critically assess the applications of present microtechnologies in the field of experimental mechanobiology; and explore opportunities and possibilities to use these tools to obtain a deeper understanding of mechanical interactions between cells and their environment.

A Hybrid Method of Application of Independent Component Analysis to In vivo (1) H MR Spectra of Childhood Brain Tumours

Independent component analysis (ICA) can automatically extract individual metabolite, macromolecular and lipid (MMLip) components from a series of in vivo MR spectra. The traditional feature extraction (FE)-based ICA approach is limited, in that a large sample size is required and a combination of metabolite and MMLip components can appear in the same independent component. The alternative ICA approach, based on blind source separation (BSS), is weak when dealing with overlapping peaks. Combining the advantages of both BSS and FE methods may lead to better results. Thus, we propose an ICA approach involving a hybrid of the BSS and FE techniques for the automated decomposition of a series of MR spectra. Experiments were performed on synthesised and patient in vivo childhood brain tumour MR spectra datasets. The hybrid ICA method showed an improvement in the decomposition ability compared with BSS-ICA or FE-ICA, with an increased correlation between the independent components and simulated metabolite and MMLip signals. Furthermore, we were able to automatically extract metabolites from the patient MR spectra dataset that were not in commonly used basis sets (e.g. guanidinoacetate). Copyright © 2011 John Wiley & Sons, Ltd.

Regulation of Heat Shock Protein 27 Phosphorylation During Microcystin-LR-induced Cytoskeletal Reorganization in a Human Liver Cell Line

Acute exposure to microcystin-LR (MC-LR) can induce the reorganization or disruption of the cytoskeleton, but proteins or enzymes correlated with this stress response have not been fully identified. Here, we report alterations to HSP27 during MC-LR-induced cytoskeletal reorganization in the human liver cell line HL7702. The cells incubated with MC-LR exhibited the rearrangement of filamentous actins and microtubules. The activity of protein phosphatase 2A was greatly decreased by MC-LR exposure. Furthermore, MC-LR markedly increased the level of HSP27 phosphorylation with the enhanced distribution of phosphorylated HSP27 to the cytoskeleton. To further determine the regulation of MC-LR-induced HSP27 phosphorylation, the activation of the MAPK superfamily was assessed. The result showed phospho-activation of p38 MAPK, JNK and ERK1/2 by MC-LR. Increases in HSP27 phosphorylation were suppressed by pretreating cells with SB203580 or SP600125, which are inhibitors of p38 MAPK or JNK, respectively. These data suggest that phosphorylated HSP27 is involved in cytoskeletal reorganization and is regulated by MAPKs, possibly as a consequence of PP2A inhibition. Moreover, the regulation of HSP27 phosphorylation may be important in MC-LR-induced cytoskeleton reassembly, which may provide helpful insights into the mechanism of MC-LR toxicity.

Microcystin-LR Induces Cytoskeleton System Reorganization Through Hyperphosphorylation of Tau and HSP27 Via PP2A Inhibition and Subsequent Activation of the P38 MAPK Signaling Pathway in Neuroendocrine (PC12) Cells

Cyanobacteria-derived microcystin-LR (MC-LR) commonly characterized as a hepatotoxin has recently been documented to show potential neurotoxicity, but the detailed neurotoxic effects of MC-LR and its mechanisms are unclear. In the present study, the neuroendocrine PC12 cell line was used to investigate whether MC-LR causes alterations of neuronal morphology and abnormalities in the phosphorylation status of cytoskeletal-associated proteins, and to elucidate the underlying mechanisms. The results showed that treatment of PC12 cells with MC-LR-triggered microtubule (MT) and actin cytoskeleton rearrangement, leading to a loss of their filamentous distribution and the display of a similar rearrangement pattern with decreased amounts of tubules or actin fibers in the cytosol and increased amounts of these structures in the cell periphery. An increase in MT tyrosination and a decrease in MT acetylation, which demonstrated MT destabilization, were also found. Moreover, MC-LR-induced hyperphosphorylation of the neural microtubule-associated protein tau, which correlated with an increase in soluble tau and a decrease in cytoskeleton-associated tau. Besides, the phosphorylation of the actin-associated protein HSP27 was also increased in MC-LR-treated cells. Furthermore, MC-LR caused a concentration-dependent decrease in the activity of protein phosphatase 2A (PP2A), and a dramatic activation of p38 mitogen-activated protein kinase (MAPK). The dephosphorylated tau dissociated from PP2A, whereas the tau phosphorylation status paralleled the activation of p38 MAPK. Pretreatment with the p38 MAPK inhibitor SB203580 effectively abolished hyperphosphorylation of tau and HSP27, and blocked MC-LR-triggered cytoskeletal alterations. Taken together, MC-LR leads to the reorganization of cytoskeletal architectures in PC12 cells and hyperphosphorylation of tau and HSP27, which may be caused by direct PP2A inhibition and indirect p38 MAPK activation.

Chemotherapeutic Properties of Phospho-nonsteroidal Anti-inflammatory Drugs, a New Class of Anticancer Compounds

Nonsteroidal anti-inflammatory drugs (NSAID) exhibit antineoplastic properties, but conventional NSAIDs do not fully meet safety and efficacy criteria for use as anticancer agents. In this study, we evaluated the chemotherapeutic efficacy of 5 novel phospho-NSAIDs, each of which includes in addition to the NSAID moiety a diethylphosphate linked through a butane moiety. All 5 compounds inhibited the growth of human breast, colon, and pancreatic cancer cell lines with micromolar potency. In vivo investigations confirmed the antitumor activity of phospho-aspirin (PA) and phospho-sulindac (PS) in inhibiting tumor growth in established human xenograft models, in which cell proliferation was suppressed and apoptosis enhanced in the absence of detectable animal toxicity. Notably, all of the phospho-NSAIDs tested induced reactive oxygen and nitrogen species in cultured cells, with PA and PS inducing detectable levels of oxidative stress in vivo that were associated positively with apoptosis and negatively with proliferation. Potentially explaining these effects, all of the phospho-NSAIDs tested also inhibited the thioredoxin system and the redox sensitive transcription factor NF-κB. Taken together, our findings show the strong anticancer efficacy and promising safety of phospho-NSAIDs in preclinical models of breast, colon, and pancreatic cancer, suggesting further evaluation as anticancer agents.

Microfluidic Approaches for Gene Delivery and Gene Therapy

Recent advances in microfluidics have created new and exciting prospects for gene delivery and therapy. The micro-scaled environment within microfluidic systems enables precise control and optimization of multiple processes and techniques used in gene transfection and the production of gene and drug transporters. Traditional non-viral gene transfection methods, such as electroporation, microinjection and optical gene transfection, are improved from the use of innovative microfluidic systems. Additionally, microfluidic systems have also made the production of many viral and non-viral vectors controlled, automated, and reproducible. In summary, the development and application of microfluidic systems are producing increased efficiency in gene delivery and promise improved gene therapy results.

HLA-DRB1 May Be Antagonistically Regulated by the Coordinately Evolved Promoter and 3'-UTR Under Stabilizing Selection

HLA-DRB1 is the most polymorphic MHC (major histocompatibility complex) class II gene in human, and plays a crucial role in the development and function of the immune system. Extensive polymorphisms exist in the promoter and 3'-UTR of HLA-DRB1, especially a LTR (Long terminal repeat) element in the promoter, which may be involved in the expression regulation. However, it remains unknown how the polymorphisms in the whole promoter region and 3'-UTR to regulate the gene expression. In this study, we investigated the extensive polymorphisms in the HLA-DRB1 promoter and 3'-UTR, and how these polymorphisms affect the gene expression in both independent and jointly manners. It was observed that most of the haplotypes in the DRB1 promoter and 3'-UTR were clustered into 4 conserved lineages (H1, H2, H3 and H4), and showed high linkage disequilibrium. Compared with H1 and H2 lineage, a LTR element in the promoter of H3 and H4 lineage significantly suppressed the promoter activity, whereas the activity of the linked 3'-UTR increased, leading to no apparent difference in the final expression product between H1/H2 and H3/H4 lineage. Nevertheless, compared with the plasmid with a promoter and 3'-UTR from the same lineage, the recombinant plasmid with a promoter from H2 and a 3'-UTR from H3 showed about double fold increased luciferase activity, Conversely, the recombinant plasmid with a promoter from H3 and a 3'-UTR from H2 resulted in about 2-fold decreased luciferase activity. These results indicate that the promoter and 3'-UTR of HLA-DRB1 may antagonistically regulate the gene expression, which may be subjected to stabilizing selection. These findings may provide a novel insight into the mechanisms of the diseases associated with HLA-DRB1 genes.

Comparing Enterovirus 71 with Coxsackievirus A16 by Analyzing Nucleotide Sequences and Antigenicity of Recombinant Proteins of VP1s and VP4s

Enterovirus 71 (EV71) and Coxsackievirus A16 (CA16) are two major etiological agents of Hand, Foot and Mouth Disease (HFMD). EV71 is associated with severe cases but not CA16. The mechanisms contributed to the different pathogenesis of these two viruses are unknown. VP1 and VP4 are two major structural proteins of these viruses, and should be paid close attention to.

Bis(η-1-tert-butyl-inden-yl)nickel(II)

The title compound, [Ni(C(13)H(15))(2)], shows a slightly distorted sandwich structure with two independent mol-ecules in the asymmetric unit. Both Ni atoms are located on crystallographic centres of inversion.

Surface-enhanced Fluorescence from Silver Fractallike Nanostructures Decorated with Silver Nanoparticles

Fluorescence emission of fluorophore molecules in the close vicinity of a nanostructured metal surface can be enhanced through a local electromagnetic field with the help of surface plasmon resonance. The fluorescence enhancement effect is very sensitive to the topography and dielectric property of the metal substrate. In the current work, metal substrates with complex structures, which are made of silver fractallike structures and nanoparticles (NPs), are prepared through electrochemical reduction followed by physical deposition. The surface-enhanced fluorescence of Rhodamine 6G monolayer molecules deposited on the prepared complex substrates are investigated with the laser spectroscopic technique. The experimental results show that the fractallike structure decorated with silver NPs presents stronger fluorescence enhancement, compared with silver NPs or pure silver fractallike structures.

[Analysis of Clinical Manifestations of Hospitalized Children Infected with Seasonal Influenza A Virus and 2009 Novel Influenza A (H1N1) Virus in Beijing]

The novel influenza A (H1N1) virus firstly detected in April 2009 in Mexico rapidly spread to many countries including the United States and Canada where humans were infected with the H1N1 virus and deaths were reported. The pandemic virus strain had never been detected in specimen of human beings and swine. It was so highly contagious and widely spread that threatened life of humans globally. This study aimed to analyze clinical data of hospitalized children patients with 2009 novel H1N1 influenza A virus infection confirmed by etiologic tests, and compared with that of seasonal influenza A.

Sagittal Alignment Comparison of Bryan Disc Arthroplasty with ProDisc-C Arthroplasty: a Prospective, Randomized Controlled Clinical Trial

A prospective, randomized study of the radiographic outcomes in patients undergoing single-level cervical arthroplasty with Bryan disc and ProDisc-C prosthesis.

Effect of the Decompressive Extent on the Magnitude of the Spinal Cord Shift After Expansive Open-door Laminoplasty

A retrospective study to analyze the effect of decompressive extent on the posterior shift of spinal cord after expansive open-door laminoplasty (ELAP).

Automated Sperm Immobilization for Intracytoplasmic Sperm Injection

Sperm immobilization is a requisite step in intracytoplasmic sperm injection (ICSI). Conventionally, sperm immobilization is performed manually, which entails long training hours and stringent skills. Manual sperm immobilization also has the limitation of low success rates and poor reproducibility due to human fatigue and skill variations across operators. This paper presents a system for fully automated sperm immobilization to eliminate limitations in manual operation. Integrating computer vision and motion control algorithms, the automated system is able to visually track a sperm and control a micropipette to immobilize the sperm. A robust sperm tail tracking algorithm is developed to locate the optimal position on the sperm tail for sperm immobilization. The system demonstrates: 1) an average sperm tail tracking error of 0.95 μm; 2) a sperm tail visual tracking success rate of 96%; 3) a sperm immobilization success rate of 88.2% (based on 1000 trials); and 4) a speed of 6-7 s per successful immobilization.

The Increased Range of Cervical Flexed Motion Detected by Radiographs in Hirayama Disease

Cervical flexion movement is supposed to play an important role in the pathogenesis of Hirayama disease. But there is no report on the range of cervical flexed motion in this disease. The purpose of current study was to compare the range of cervical flexed motion in patients with Hirayama disease with the one in healthy controls using conventional lateral flexion radiographs of the cervical spine, and to investigate the diagnostic value of radiographs for Hirayama disease.

Demographic Characteristics and Prevalence of Serologic Markers Among Blood Donors Who Use Confidential Unit Exclusion (CUE) in São Paulo, Brazil: Implications for Modification of CUE Policies in Brazil

This study evaluated demographic profiles and prevalence of serologic markers among donors who used confidential unit exclusion (CUE) to assess the effectiveness of CUE and guide public policies regarding the use of CUE for enhancing safety versus jeopardizing the blood supply by dropping CUE.

A Fast and Simple Method to Fabricate Circular Microchannels in Polydimethylsiloxane (PDMS)

A simple method to fabricate circular microchannels in polydimethylsiloxane (PDMS) is presented. A coating of liquid PDMS is applied on the walls of rectangular microchannels, fabricated using standard soft-lithography, by introducing a pressurized air stream inside the PDMS filled microchannels. Surface tension of the liquid PDMS forces the coating to take a circular cross-section which is preserved by baking the device to cure the coated layer. Diameters ranging from a few micrometres to a few hundreds of micrometres were achieved. The method was verified to work on microchannel networks as well as in straight channels. Different coating conditions were systematically tested. Design curves are reported for one to choose appropriate coating conditions for obtaining a desired diameter. A comparison between the performance of square and circular microchannels in trapping SiHa cells (cervical cancer cell line) is shown.

A Superficial Colon Tumor Model Involving Subcutaneous Colon Translocation and Orthotopic Transplantation of Green Fluorescent Protein-expressing Human Colon Tumor

The orthotopic transplantation model of human tumor has been demonstrated to be more patient-like animal tumor model. However, observations of tumor progression and metastasis are limited by the deep location of the colon or limited deep penetration ability of fluorescence through tissue. The purpose of this study is to establish a superficial orthotopic model to allow easier real-time visualization and more sensitive monitoring of fluorescent orthotopic colon tumor. Human colon cancer HT-29 cells were transduced with a pLPCX expression retroviral vector containing green fluorescent protein and neomycin resistance genes. For superficial orthotopic transplantation model, the cecum was identified and pulled out of the peritoneal cavity, the space between the cecum and peritoneum was sutured, the cecum was pulled to subcutaneous tissue, and incision was made on the cecal serosa followed by the implantation of a 1-mm tumor tissue to the cecum. For comparison, a conventional orthotopic transplantation model was established in a separate group of mice simultaneously. When tumor sizes reached 5 mm in diameter, half the mice in each model received 5-FU treatment. Primary tumor and metastases were monitored by fluorescent imaging or caliber measurement. Tumor fluorescence was observed as early as 3 days (median time of 4.7 ± 1.3 days) post-transplantation in the superficial orthotopic transplantation model, which was much earlier than 21 days (median time of 26.2 ± 9.9 days) in conventional orthotopic transplantation model. Although tumor growth of 5-FU-treated mice in conventional orthotopic model was lower than those of the untreated mice, the difference was not significant. However, in superficial orthotopic model, tumor growth was significantly inhibited in 5-FU-treated mice relative to the untreated mice. Fluorescence imaging showed similar metastasis incidence between the superficial and conventional orthotopic transplantation models. The fluorescent superficial orthotopic transplantation colon model allows easier real-time visualization and more sensitive monitoring of tumor growth as well as convenient repeated sampling. It is a valuable orthotopic implantation model for study of colon cancer and evaluation of new anti-cancer therapy.

Agreement Among HLA Antibody Detection Assays is Higher in Ever-pregnant Donors and Improved Using a Consensus Cutoff

HLA antibodies might contribute to the pathogenesis of transfusion-related acute lung injury (TRALI). HLA antibody detection methods include ELISA, flow cytometry, and multiplex bead-based assays, as well as the older lymphocytotoxicity assay, and it is not obvious how to compare results across platforms.

Increased Mitochondrial DNA Damage and Decreased Base Excision Repair in the Auditory Cortex of D-galactose-induced Aging Rats

Aging has been associated with mitochondrial DNA (mtDNA) common deletion (CD). Age changes in the central auditory system are well known to affect speech perception. Base excision repair (BER) is the major type of DNA repair in mitochondria. The current study was designed to investigate potential causative mechanisms of central presbycusis by using a rat mimetic aging model induced by subcutaneous administration of D-galactose (D-gal). Quantitative real-time PCR and Western blotting analyses were performed to identify the mtDNA 4834 bp deletion and selected mitochondrial DNA repair enzymes, DNA polymerase γ (pol γ) and 8-oxoguanine DNA glycosylase (OGG1). Cell apoptosis in the auditory cortex was detected using terminal deoxynucleotidyltransferase mediated UTP nick-end labeling (TUNEL). Our data showed that mtDNA 4834 bp deletion and TUNEL-positive cells were significantly increased and the expression of pol γ and OGG1 were remarkably down-regulated in the auditory cortex in D-gal-treated rats compared to control rats. During aging, increased mtDNA damage likely results from decreased DNA repair capacity in the auditory cortex. DNA repair enzymes such as pol γ and OGG1 may provide novel pharmacological targets to promote DNA repair and rescue the central auditory system in patients with degenerative diseases.

Genetic Variation Associated with Bortezomib-induced Peripheral Neuropathy

To develop a predictive genetic signature for the development of bortezomib-induced peripheral neuropathy (PN).

Macroscopic Investigation of the Transient Hydrodynamic Memory Behavior of Preparative Packed Chromatography Beds

The goal of the present work was to examine the hydrodynamic behavior of preparative scale packed chromatography beds during long-term cyclical operation at high loads using an experimental set-up with a high resolution measuring device of bed height. One agarose-based resin and one methacrylic-based resin were examined in a 140 mm column. Both resins exhibited hysteresis behavior during compression/relaxation cycles. The hystereses were less pronounced with decreasing hydrodynamic stress rate. The occurrence of hystereses was an indication for hydrodynamic memory behavior of the chromatography packing. During long-term cyclical operation at high loads of the column filled with methacrylic resin, oscillations of the steadily with time decreasing flow rate were observed for the first time. These oscillations were attributed to the viscoelasticity of the polymer particles network representing a system with materials with fading memory. Such nonlinear systems with feed-back are known to exhibit inherent self-oscillations. A decoupling of the two processes of bed compression and decrease of bed permeability was observed. The presented results explain why preparative packed-bed chromatography often yields unsatisfactory reproducible data and why unwanted phenomena like medium wall detachment or other symptoms of deteriorated chromatography beds are frequently observed.

An Automated Microfluidic Sample Preparation System for Laser Scanning Cytometry

Laser scanning cytometry (LSC) is emerging as a clinical tool. In one application a "Clatch" slide, named after the inventor, is used in conjunction with LSC for cell surface marker immunophenotyping of patient samples. The slide requires time consuming and laborious pipetting steps, making a test tedious and prone to handling errors. The Clatch slide also uses a significant number of cells, limiting the number of analyses on paucicellular samples. This paper presents an automated microfluidic system consisting of a control circuit, a microfluidic system, and an aluminum frame, capable of performing immunophenotyping procedures. This prototype system reduces 36 pipetting steps to 1, reduces the amount of cell sample from 180 μL to 56 μL, and shortens the time used by technicians.

Efficiency of Anthropometric Indicators of Obesity for Identifying Cardiovascular Risk Factors in a Chinese Population

To evaluate the predictive value of the body mass index (BMI), waist circumference (WC), and waist-to-height ratio (WHtR) for the presence of several cardiovascular risk conditions-hypertension, dyslipidaemia, metabolic syndrome (MS), and type 2 diabetes-in a Chinese population in Jinan, China.

Innate Immune Function of the Adherens Junction Protein P120-catenin in Endothelial Response to Endotoxin

Sepsis-induced acute lung injury is a common clinical disorder in critically ill patients that is associated with high mortality. In this study, we investigated the role of p120-catenin (p120), a constituent of endothelial adherens junctions, in regulating the innate immune function of lungs. In mice in which acute lung injury was induced by i.p. administration of LPS, we observed a rapid decrease in the expression of p120 in lungs. The p120 protein expression was correlated inversely with severity of inflammation. Suppression of p120 expression in lung endothelial cells in mice using small interfering RNA resulted in high sensitivity to endotoxin and greatly increased the mortality compared with controls. Knockdown of p120 also increased the expression of ICAM-1, neutrophil recruitment, production of cytokines TNF-α and IL-6, pulmonary transvascular protein permeability, and lung water content in response to LPS. We demonstrated that endothelial p120 modulates lung innate immune function by interfering with the association of TLR4 with its adaptor MyD88 to block TLR4 signaling and NF-κB activation in endothelial cells. In conclusion, these studies have uncovered a novel innate immune function of endothelial p120 in downregulating the lung inflammatory response to endotoxin through the suppression of TLR4 signaling.

Methylation-mediated Regulation of E2F1 in DNA Damage-induced Cell Death

E2F1 promotes DNA damage-induced apoptosis and the post-translational modifications of E2F1 play an important role in the regulation of E2F1-mediated cell death. Here, we found that Set9 and LSD1 regulate E2F1-mediated apoptosis upon DNA damage. Set9 methylates E2F1 at lysine 185, a conserved residue in the DNA-binding domain of E2F family proteins. The methylation of E2F1 by Set9 leads to the stabilization of E2F1 and up-regulation of its proapoptotic target genes p73 and Bim, and thereby induces E2F1-mediated apoptosis in response to genotoxic agents. We also found that LSD1 demethylates E2F1 at lysine 185 and reduces E2F1-mediated cell death. The identification of the methylation/demethylation of E2F1 by Set9/LSD1 suggests that E2F1 is dynamically regulated by epigenetic enzymes in response to DNA damage.

[Digital Inlaid Design Based on Anatomical Characteristics and Preliminary Implementation of Its Program]

To explore the technical guideline of inlays computer aided design (CAD) for further development, to test the feasibility of processing for the design results of the technical guideline, and to assist co-operation with automatic inlay CAD software development.

Establishing Assay Cutoffs for HLA Antibody Screening of Apheresis Donors

Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion-related deaths. Donor HLA antibodies have been implicated in TRALI cases. Blood centers are implementing TRALI risk reduction strategies based on HLA antibody screening of some subpopulations of ever-pregnant apheresis platelet (PLT) donors. However, if screening assay cutoffs are too sensitive, donation loss may adversely impact blood availability.

Application of GFAT As a Novel Selection Marker to Mediate Gene Expression

The enzyme glutamine: fructose-6-phosphate aminotransferase (GFAT), also known as glucosamine synthase (GlmS), catalyzes the formation of glucosamine-6-phosphate from fructose-6-phosphate and is the first and rate-limiting enzyme of the hexosamine biosynthetic pathway. For the first time, the GFAT gene was proven to possess a function as an effective selection marker for genetically modified (GM) microorganisms. This was shown by construction and analysis of two GFAT deficient strains, E. coli ΔglmS and S. pombe Δgfa1, and the ability of the GFAT encoding gene to mediate plasmid selection. The gfa1 gene of the fission yeast Schizosaccharomyces pombe was deleted by KanMX6-mediated gene disruption and the Cre-loxP marker removal system, and the glmS gene of Escherichia coli was deleted by using λ-Red mediated recombinase system. Both E. coli ΔglmS and S. pombe Δgfa1 could not grow normally in the media without addition of glucosamine. However, the deficiency was complemented by transforming the plasmids that expressed GFAT genes. The xylanase encoding gene, xynA2 from Thermomyces lanuginosus was successfully expressed and secreted by using GFAT as selection marker in S. pombe. Optimal glucosamine concentration for E. coli ΔglmS and S. pombe Δgfa1 growth was determined respectively. These findings provide an effective technique for the construction of GM bacteria without an antibiotic resistant marker, and the construction of GM yeasts to be applied to complex media.

New Hexanuclear Group 11 Pyrazolate Complexes: Synthesis and Photophysical Features

The treatment of 2,2'-di(1,2-pyrazol-3-yl)-1,1'-binaphthyl with two equivalents of appropriate monovalent group 11 precursors in the presence of a base leads to a complete exchange of the pyrazole NH protons with M(+) cations. Structural characterisation of the copper(I) complex revealed a hexanuclear complex with a pelton-wheel-like arrangement of the binaphthyl unit. As indicated by their spectroscopic data, all three complexes are isostructural. The complexes show a complex fluorescence behaviour that can be partially related to the ligand system and also to the metal sites, as indicated by the position of the fluorescence peaks and their temperature dependence and lifetimes.

Genetic Polymorphisms of Nerve Growth Factor Receptor (NGFR) and the Risk of Alzheimer's Disease

ABSTRACT: BACKGROUND: Loss of basal forebrain cholinergic neurons is attributable to the proapoptotic signaling induced by nerve growth factor receptor (NGFR) and may link to Alzheimer's disease (AD) risk. Only one study has investigated the association between NGFR polymorphisms and the risk of AD in an Italian population. Type 2 diabetes mellitus (DM) may modify this association based on previous animal and epidemiologic studies. METHODS: This was a case-control study in a Chinese population. A total of 264 AD patients were recruited from three teaching hospitals between 2007 to 2010; 389 controls were recruited from elderly health checkup and volunteers of the hospital during the same period of time. Five common (frequency>=5%) haplotype-tagging single nucleotide polymorphisms (htSNPs) were selected from NGFR to test the association between NGFR htSNPs and the risk of AD. RESULTS: Variant NGFR rs734194 was significantly associated with a decreased risk of AD [GG vs. TT copies: adjusted odds ratio (OR)=0.43, 95% confidence interval (CI)=0.20-0.95]. Seven common haplotypes were identified. Minor haplotype GCGCG was significantly associated with a decreased risk of AD (2 vs. 0 copies: adjusted OR=0.39, 95% CI=0.17-0.91). Type 2 DM significantly modified the association between rs2072446, rs741072, and haplotype GCTTG and GTTCG on the risk of AD among ApoE 4 non-carriers (Pinteraction<0.05). CONCLUSION: Inherited polymorphisms of NGFR were associated with the risk of AD; results were not significant after correction for multiple tests. This association was further modified by the status of type 2 DM.

A Rotating Disk Electrode Study of the Particle Size Effects of Pt for the Hydrogen Oxidation Reaction

By using a catalyst-lean thin-film RDE method, the fast kinetics of the hydrogen oxidation reaction (HOR) on highly dispersed Pt nanoparticle electrocatalysts can be determined, free from the interference of the mass transport of H(2) molecules in solution. Measurements with carbon-supported Pt nanoparticles of different sizes thus allow revealing the particle size effect of Pt for the HOR. It is shown that there is a "negative" particle size effect of Pt on the kinetics of HOR, i.e., the exchange current density j(0) decreases with the increased dispersion (i.e. decreased mean particle size). A maximum mass activity of Pt for the HOR is found at particle sizes of 3-3.5 nm. The observed particle size effect is interpreted in terms of the size dependent distribution of surface atoms on the facets and edges, which is implied by the voltammetric responses of Pt/C catalysts with differently sized Pt particles. The accompanied decrease in the HOR activity with the increase in the edge atom fraction suggests that the edge atoms on the surface of Pt nanoparticles are less active for the HOR than those on the facets.

A New Alternative NF-ΚB Pathway Mediated the Neuroprotection of GDNF on 6-OHDA-induced DA Neurons Neurotoxicity

Glial cell line-derived neurotrophic factor (GDNF) is a potent protective factor for dopaminergic (DA) neurons, but the signaling mechanisms underlying the effect of GDNF on these neurons remain obscure. Here, both our in vivo and in vitro studies demonstrate that the majority of DA neurons express the NF-κB-inducing kinase (NIK), which is the essential kinase for mediating activation of the new alternative NF-κB signaling pathway. Additionally, we also show that GDNF induced the time/dose-dependent phosphorylation of IκB kinase α (IKKα) and p100, facilitated the processing of p100 to p52 and accelerated the translocation of NF-κB dimmers into the nuclei of DA neurons. We furtherly found that the dimmer which translocate into the nucleus was RelA/p52 not RelB/p52. Meanwhile, the attenuation of 6-OHDA-induced DA neuronal apoptosis due to GDNF was reversed subsequent to the inhibition of p100 expression by RNAi while the neuroprotective effect of GDNF on injured DA neurons was strengthened by the overexpression of p100. Our data, therefore, indicate that a new alternative NF-κB signaling pathway, which was not the classic pathway but different from the non-canonical pathway, exists in DA neurons and mediates the neuroprotective effect of GDNF on these neurons.

How Does the Neck Flexion Affect the Cervical MRI Features of Hirayama Disease?

Although flexion cervical MRI has been recommended for the diagnosis of Hirayama disease (HD), no study focused on the MR features at different neck flexion angles. Moreover, no uniform flexion angle has been confirmed in clinical practice. The purpose of this study is to quantitatively investigate the MRI typical signs of HD patients in different neck flexion degree and gives a suggestion to the MR scanning. Cervical MRI in neutral and different flexion positions (cervical flexion angle 20°, 25°, 30°, 35°, and 40°) were performed in 45 HD patients. Three MRI features including anterior shifting of the posterior wall of the cervical dural canal (ASD), widening of cervical epidural space, and epidural flow voids (EFV) at each flexed position were summarized. To evaluate ASD quantitatively, the widest cervical epidural space with the maximum sagittal diameters (d) and cervical canal sagittal diameter (D) at the same level were measured. The d/D values at different angles were calculated and compared. ASD was demonstrated in 34 out of 45 cases (75.6%) at 20° and in all cases (100%) at other 4 angles (χ (2) = 25.728, P < 0.05). Significant difference was demonstrated for the appearance rate of EFV (mean 72.8%) among different angles (χ (2) = 11.373, P = 0.021). The peak mean d/D value was found at 35°. Neck flexion angles have effects on ASD, widening of cervical epidural space and EFV. 25° is recommended as the least effective diagnostic flexion angle for MRI diagnosis of HD, and 35° may be the best one.

Comparison of Annual Versus Twice-yearly Mass Azithromycin Treatment for Hyperendemic Trachoma in Ethiopia: a Cluster-randomised Trial

In trachoma control programmes, azithromycin is distributed to treat the strains of chlamydia that cause ocular disease. We aimed to compare the effect of annual versus twice-yearly distribution of azithromycin on infection with these strains.

A Comparative Study of the Preventive Effects of Mitomycin C and Chitosan on Intraarticular Adhesion After Knee Surgery in Rabbits

We sought to compare the preventive effects of mitomycin-C(MMC) and chitosan on intraarticular adhesion after knee surgery in rabbits. For this purpose, 48 New-Zealand rabbits were randomly and equally divided into MMC, chitosan, and control groups. Approximately 10 × 10 mm(2) of the cortical bone was removed from both sides of left femoral condyle and the cancellous bone underneath was exposed. The decorticated areas were topically treated with MMC and chitosan while control group was treated with physiological saline. The lower left limb was fixed in flexed position with Kirschner-wire for 4 weeks postoperatively. After 4 weeks, gross and histopathological examination, biochemical analysis, and fibroblast counts were performed on knee intraarticular adhesion in each group. The data show mild membrane-like fibrous intraarticular adhesion, presented in loose, in MMC group. There was moderate intraarticular adhesion in chitosan group while in controls; there was large-size compact fibrous tissue adhesion. Hydroxyproline contents and fibroblast quantity of MMC and chitosan groups were lower (P < 0.05) than that of control group. We, therefore, concluded that MMC and chitosan could prevent intraarticular adhesion of the knee in rabbits by inhibiting fibroblast proliferation and reducing collagenous fiber formation while MMC had a better preventive effect than that of chitosan.

Effect of Oscillating Fluid Flow Stimulation on Osteocyte MRNA Expression

Structural adaptation of the bone tissue is mediated by loading-induced interstitial fluid flow within the bone microstructure. Within this framework, osteocytes fulfill the central mechanotransductive role in the bone remodeling process. While osteocytes have been demonstrated to be exquisitely sensitive to various forms of fluid flow stimulus in vitro, the effect of different oscillating fluid flow (OFF) parameters on osteocyte activity has yet to be systematically characterized. In this study, we investigate the effect of three OFF parameters on osteocyte activity in vitro and hypothesize that COX-2, RANKL, and OPG mRNA expression in osteocytes are sensitive to the OFF parameters: peak shear stress amplitude (0.5 Pa, 1 Pa, 2 Pa, and 5 Pa), oscillating frequency (0.5 Hz, 1 Hz, and 2 Hz), and total flow duration (1 h, 2 h, and 4 h). Our findings demonstrate that COX-2 mRNA levels are elevated in osteocytes subjected to higher peak shear stress amplitudes and longer flow durations, while RANKL/OPG mRNA levels decreased to a minimum threshold in response to higher peak shear stress amplitudes, faster oscillating frequencies, and longer flow durations. These findings suggest that dynamic fluid flow with higher peak shear stress amplitudes, faster oscillating frequencies, and longer loading durations provide the best conditions for promoting bone formation.

Sigma-1 Receptor Agonists Provide Neuroprotection Against Gp120 Via a Change in Bcl-2 Expression in Mouse Neuronal Cultures

Although combined antiretroviral therapy has significantly improved the prognosis of HIV-1 infected patients and decreased the incidence of HIV-1 associated dementia, the cumulative prevalence of this disease, in particular, mild or asymptomatic neurocognitive impairment, has not decreased. Thus, in addition to active antiretroviral therapy, the search for an effective neuroprotective approach is very important. Sigma-1 receptors are widely distributed in the central nervous system. Sigma-1 receptor agonists are robustly neuroprotective in many neuropathy and neurotoxicity in vivo and in vitro studies. This study aims to investigate possible neuroprotective effects of sigma-1 receptor agonist, 4-phenyl-1-(4-phenylbutyl) piperidine (PPBP) against HIV-1 protein gp120. Primary cortical neuronal cultures were exposed to gp120 in different concentrations; to investigate neuroprotective effects of sigma-1 receptor agonist, cells were pre-treated with PPBP (10μM) in the presence or absence of pre-incubated sigma-1 receptor antagonist rimcazole (5μM). Cell apoptosis was confirmed with calcein/PI uptake test, lactate dehydrogenase (LDH) leakage assay or TUNEL assay and neurite degeneration was evaluated with morphometry via MAP-2 stained immunofluorescence. The mRNA and protein levels of apoptosis associated bax and bcl-2 were determined with real-time qPCR and Western blot. The results showed that gp120 could induce neuronal apoptosis and neurite degeneration in a concentration dependent manner and PPBP could attenuate the neurotoxicity of gp120. Simultaneously, gp120 could induce low expression of bcl-2 and bax, but only low expression of bcl-2 could be reversed by PPBP. The present data suggest that PPBP, at least, in part protects the neuron against gp120 by regulating bcl-2 expression.

Hypoxia Inducible Factor-1α is Involved in the Neurodegeneration Induced by Isoflurane in the Brain of Neonatal Rats

More and more data show isoflurane, a commonly used volatile anesthetic has dual effects on neuron fate. However, the underlying mechanisms that can explain the apparent paradox are poorly understood. Hypoxia inducible factor (HIF)-1α, a transcription factor, has been found regulating both prosurvival and prodeath pathways in the CNS. Previously, we found that isoflurane can activate HIF-1α under normoxic conditions in vitro and HIF-1α has been found to be involved in the pre-conditioning effect of isoflurane in various organs. Here, we investigated whether HIF-1α is a contributing factor in the neurodegenration in rodent primary cultured neurons and in developing rat brain. Isoflurane dose-dependently induced apoptotic neurodegeneration in neonatal rats as assessed by S100β, cleaved caspase 3 and poly-(ADP-ribose) polymerase (PARP), respectively. Notably, isoflurane up-regulates HIF-1α protein levels in vivo and in vitro during induction of neurodegeneration. Likewise, isoflurane resulted in a significant elevation of cytosonic calcium levels in neuron cultures. Furthermore, knockdown of HIF-1α expression in cultured neurons attenuated isoflurane-induced neurotoxicity. Finally, Morris water maze (MWM) test showed neonatal exposure to isoflurane impaired juvenile learning and memory ability in rats. These findings indicate that HIF-1α is involved in the neurodegeneration induced by isoflurane in the brain of neonatal rats, suggesting HIF-1α may be a candidate for the dual effects of isoflurane on neuron fate.

Sequence Variants of Interleukin 6 (IL-6) Are Significantly Associated with a Decreased Risk of Late-onset Alzheimer's Disease

ABSTRACT: BACKGROUND: Interleukin 6 (IL-6) has been related to beta-amyloid aggregation and the appearance of hyperphosphorylated tau in Alzheimer's disease (AD) brain. However, previous studies relating IL-6 genetic polymorphisms to AD included few and unrepresentative single nucleotide polymorphisms (SNPs) and the results were inconsistent. METHODS: This is a case-control study. A total of 266 patients with AD, aged65, were recruited from three hospitals in Taiwan (2007-2010). Controls (n = 444) were recruited from routine health checkups and volunteers of the hospital during the same period of time. Three common IL-6 haplotype-tagging SNPs were selected to assess the association between IL-6 polymorphisms and the risk of late-onset AD (LOAD). RESULTS: Variant carriers of IL-6 rs1800796 and rs1524107 were significantly associated with a reduced risk of LOAD [(GG + GC vs. CC): adjusted odds ratio (AOR) = 0.64 and (CC + CT vs. TT): AOR = 0.60, respectively]. Haplotype CAT was associated with a decreased risk of LOAD (0 and 1 copy vs. 2 copies: AOR = 0.65, 95% CI = 0.44-0.95). These associations remained significant in ApoE e4 non-carriers only. Hypertension significantly modified the association between rs2069837 polymorphisms and the risk of LOAD (pinteraction = 0.03). CONCLUSIONS: IL-6 polymorphisms are associated with reduced risk of LOAD, especially in ApoE e4 non-carriers. This study identified genetic markers for predicting LOAD in ApoE e4 non-carriers.

OTUB1 Overexpression in Mesangial Cells Is a Novel Regulator in the Pathogenesis of Glomerulonephritis Through the Decrease of DCN Level

OTUB1 is a member of OTUs (Ovarian-tumor-domain-containing proteases), a deubiquitinating enzymes family (DUBs), which was shown as a proteasome-associated DUB to be involved in the proteins Ub-dependent degradation. It has been reported that OTUB1 was expressed in kidney tissue. But its concrete cellular location and function in the kidney remain unclear. Decorin (DCN) in mesangial cells (MC) is considered to be a potentially important factor for antagonizing glomerulonephritides, and its degradation is mediated by ubiquitination. The aim of this study is to investigate the role of OTUB1 expression in MC and its relationship with DCN during glomerulonephritis.

Genetic Polymorphisms of Clusterin Gene Are Associated with a Decreased Risk of Alzheimer's Disease

Reliability of Measurements Performed by Community-drawn Anthropometrists from Rural Ethiopia

Undernutrition is an important risk factor for childhood mortality, and remains a major problem facing many developing countries. Millennium Development Goal 1 calls for a reduction in underweight children, implemented through a variety of interventions. To adequately judge the impact of these interventions, it is important to know the reproducibility of the main indicators for undernutrition. In this study, we trained individuals from rural communities in Ethiopia in anthropometry techniques and measured intra- and inter-observer reliability.

Phospho-ibuprofen (MDC-917) Suppresses Breast Cancer Growth: an Effect Controlled by the Thioredoxin System

ABSTRACT: INTRODUCTION: We have recently synthesized phospho-ibuprofen (MDC-917; P-I), a safer derivative of ibuprofen, which has shown anticancer activity. We investigated its efficacy and mechanism of action in the treatment of breast cancer in preclinical models. METHODS: We evaluated the anti-breast cancer efficacy of P-I alone or incorporated into liposomes (Lipo-P-I) in human ER (+) (MCF-7) and triple-negative (ER(-), PgR(-) and HER2(-); MDA-MB231) breast cancer cell lines, as they represent the most frequent (ER(+)) and the most difficult to treat (triple-negative) subtypes of breast cancer , and their xenografts in nude mice. We assessed the effect of P-I on a) the levels of reactive oxygen nitrogen species in response to P-I using molecular probes; b) the thioredoxin system (expression and redox status of thioredoxin-1 (Trx-1) and thioredoxin reductase activity); c) cyclooxygenase 2 (COX-2), nuclear factor KB (NF-KB) and mitogen-activated protein kinase (MAPK) cell signaling; and d) the growth of xenografts with stably knocked-down Trx-1. RESULTS: Compared to controls, a) P-I 400 mg/kg/day inhibited the growth of MDA-MB231 xenografts 266%; and b) P-I 300 mg/kg/day inhibited the growth of MCF-7 xenografts 51% and Lipo-P-I at the same dose 181%. In both cell lines, P-I induced oxidative stress and suppressed the Trx system (oxidized Trx-1 and decreased its expression; inhibited thioredoxin reductase activity). These changes triggered downstream redox signaling: the activity of NF-KB was suppressed and the Trx-1-ASK1 complex was dissociated, activating the p38 and JNK MAPK cascades. Trx-1 knockdown abrogated P-I's anticancer effect in vitro and in vivo. CONCLUSIONS: P-I is safe and effective against breast cancer. Liposomal formulation enhances its efficacy; its effect is heavily dependent on the induction of oxidative stress and the suppression of the thioredoxin system. P-I merits further evaluation as an agent for the treatment of breast cancer.

Polymorphisms of ICAM-1 Are Associated with Gastric Cancer Risk and Prognosis

To investigate the association between single nucleotide polymorphisms (SNPs) in intercellular adhesion molecule-1 (ICAM-1) and the risk, biological behavior and prognosis of gastric cancer (GC) in Chinese population.

How Big is a Cp? Cycloheptatrienyl Zirconium Complexes with Bulky Cyclopentadienyl and Indenyl Ligands

A combination of phase-transfer and traditional alkylation strategies has been employed to synthesise sterically encumbered 1,3-di(cyclohexyl) and 1,3-di(tert-butyl) substituted indenes in multi-gram quantities. These indenyl ligands and sterically demanding alkyl cyclopentadienyl ligands have been used to prepare a series of [(η(7)-C(7)H(7))Zr(η(5)-L)] (L = Cp and Ind) complexes by straightforward salt metathesis between [(η(7)-C(7)H(7))ZrCl(tmeda)] and the corresponding sodium indenide or cyclopentadienide. All of these Zr complexes have been characterized by elemental analysis, NMR spectroscopy and single crystal X-ray diffraction. The structural information derived from these studies was employed to evaluate the steric demand of these ligands in a realistic manner.

Finite Element Analysis of Cervical Arthroplasty Combined With Fusion Against 2-Level Fusion

STUDY DESIGN:: A biomechanical analysis of cervical arthroplasty and fusion using the finite element method. OBJECTIVE:: The purpose of this study was to compare the biomechanical performances of hybrid surgery (HS, C45Fusion combined with C56ProDisc-C arthroplasty) and 2-level fusion (TLF). SUMMARY OF BACKGROUND DATA:: Cervical disk arthroplasty gained reliable clinical outcomes for treating single-level and 2-level cervical spondylosis. Cervical disk arthroplasty combined with fusion (HS) may be an alternative to 2-level anterior cervical decompression and fusion. METHODS:: The HS model and the TLF model were analyzed using the finite element method. The range of motion (ROM) and adjacent intradikcal pressures (IDPs) under flexion, extension, lateral bending, and axial rotation were calculated and compared for both models. RESULTS:: (1) Compared with the normal model, the ROM of C56 increased by 53.2% in flexion-extension, 69.3% in axial rotation, and 69.8% in lateral bending of ProDisc-C arthroplasty in the HS model. (2) The ROM of C3-C7 in the HS model was 22.9 degrees in flexion-extension, decreased by 18.9%, whereas the ROM of C3-C7 in the TLF model was 17.0 degrees, decreased by 39.7% compared with the normal model. (3) The maximal IDP of TLF model increased by 44.4% at C34 and 40.6% at C67 in flexion, whereas the HS model increased by 5.4% and 9.5%, respectively, compared with the normal model. (4) The ROM of the adjacent segment in TLF increased by 0.1% of C34 and 8.3% of C67 in flexion-extension, whereas that of the HS model decreased by 8.1% of C34 and 2.1% of C67 compared with the normal model. CONCLUSIONS:: (1) The ROM of C56 (ProDisc-C arthroplasty) in HS was increased. (2) The HS model has a better ROM of C3-C7 than the TLF model. (3) The HS model offered less increase of adjacent segmental IDP and ROM than the TLF model.

Automated Nanomanipulation for Nanodevice Construction

Nanowire field-effect transistors (nano-FETs) are nanodevices capable of highly sensitive, label-free sensing of molecules. However, significant variations in sensitivity across devices can result from poor control over device parameters, such as nanowire diameter and the number of electrode-bridging nanowires. This paper presents a fabrication approach that uses wafer-scale nanowire contact printing for throughput and uses automated nanomanipulation for precision control of nanowire number and diameter. The process requires only one photolithography mask. Using nanowire contact printing and post-processing (i.e. nanomanipulation inside a scanning electron microscope), we are able to produce devices all with a single-nanowire and similar diameters at a speed of ∼1 min/device with a success rate of 95% (n = 500). This technology represents a seamless integration of wafer-scale microfabrication and automated nanorobotic manipulation for producing nano-FET sensors with consistent response across devices.

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