VEGF gene silencing by cytomegalovirus promoter driven ShRNA expression vector results in vascular development defects in zebrafish

Zebrafish has been generally considered as an excellent model in case of drug screening, disease model establishment, and vertebrate embryonic development study. In this work, the ability of human cytomegalovirus immediate early promoter (CMV promoter)-driven short hairpin RNA (shRNA) expression vector to induce shRNA against VEGF gene in zebrafish was tested, and its effect on vascular development was assed, too. Using RT-qPCR, blood vessel staining, and in situ hybridization, we confirmed certain transcriptional activity and down regulation of gene expression by the vector. In situ hybridization analysis indicated selective inhibition of NRP1 expression in the VEGF gene loss of function model, which might imply in turn that VEGF could not only activate endothelial cells directly but also could contribute to stimulating angiogenesis in vivo by a mechanism that involved up-regulation of its cognate receptor expression in zebrafish. This contributed to a better understanding of molecular mechanisms of cardiovascular development. The system improved the success rate in making inducible knockdown and widened the possibilities for better therapeutic targets in zebrafish.

The development of an inhomogeneous damage field in an elastic-plastic model

By making use of the evolution equation of the damage field as derived from the statistical mesoscopic damage theory, we have preliminarily examined the inhomogeneous damage field in an elastic-plastic model under constant-velocity tension. Three types of deformation and damage field evolution are presented. The influence of the plastic matrix is examined. It seems that matrix plasticity may defer the failure due to damage evolution. A criterion for damage localization is consistent with the numerical results.

Failure process analysis of cement under explosive loading with a generalized driven nonlinear threshold model

The microstructural heterogeneity and stress fluctuation play important roles in the failure process of brittle materials. In this paper, a generalized driven nonlinear threshold model with stress fluctuation is presented to study the effects of microstructural heterogeneity on continuum damage evolution. As an illustration, the failure process of cement material under explosive loading is analyzed using the model. The result agrees well with the experimental one, which proves the efficiency of the model.

Development And Application Of Flexible Substrate Sensors In Instantaneous Heat Flux Measurement

A new type of sensor with the flexible substrate is introduced. It is applicable in measuring instantaneous heat flux on the model surface in a hypersonic shock tunnel. The working principle, structure and manufacture process of the sensor are presented. The substrate thickness and the dynamic response parameter of the sensor are calculated. Because this sensor was successfully used in measuring the instantaneous heat flux on the surface of a flat plate in a detonation-driven shock tunnel, it may be effective in measuring instantaneous heat flux on the model surface.

Theoretical model for a periodically driven semiconductor laser subject to optical feedback from a microcantilever

We propose a theoretical model for analyzing the dynamics of a periodically driven semiconductor laser subject to optical feedback from a microcantilever. We numerically investigate the temporal evolution of the light intensity of the semiconductor laser, and we show the interspikes of the light intensity. These interspikes of light intensity are also demonstrated in our experiment. The validity of the theoretical model is verified. The observed phenomenon has a potential application for resonant sensing. (C) 2008 Optical Society of America.

scenario-focused development method for a pen-based user interface: model and applications

Pen-based user interface (PUI) has drawn significant interest, owing to its intuitiveness and convenience. While much of the research focuses on the technology, the usability of a PUI has been relatively low since human factors have not been considered sufficiently. Scenario-centric designs are ideal ways to improve usability. However, such designs possess some problems in practical use. To cope with these design issues, the concept of “interface scenarios” is proposed in to facilitate the interface design, and to help users understand the interaction process in such designs. The proposed scenario-focused development method for PUI is coupled with a practical application to show its effectiveness and usability.

Rotation-driven prolate-to-oblate shape phase transition in W-190: A projected shell model study

A shape phase transition is demonstrated to occur in W-190 by applying the projected shell model, which goes beyond the usual mean-field approximation. Rotation alignment of neutrons in the high-j, i(13/2) orbital drives the yrast sequence of the system, changing suddenly from prolate to oblate shape at angular momentum 10h. We propose observables to test the picture.

Suppression of zebrafish VEGF gene by cytomegalovirus promoter-driven short hairpin constructs induces vascular development defects and down regulation NRP1 expression

The usage of RNA interference for gene knockdown in zebrafish through expression of the small interfering RNA mediators from DNA vectors has created a lot of excitement in the research community. In this work, the ability of human cytomegalovirus immediate early promoter (CMV promoter)-driven short hairpin RNA (shRNA) expression vector to induce shRNA against vascular endothelial growth factor (VEGF) gene in zebrafish was tested, and its effects on VEGF-mediated vasculogenesis and angiogenesis were evaluated. Altogether four vectors targeting various locations of VEGF gene were constructed, and pSI-V4 was proven to be the most effective one. Microinjection of pSI-V4 into the zebrafish embryos resulted in defective vascular formation and down regulation of VEGF expression. In situ hybridization analysis indicated that silencing VEGF gene expression by pSI-V4 resulted in down regulation of neuropilin-1 (NRP1), a potent VEGF receptor. Knockdown of VEGF expression by morpholino gave the same result. This provided evidence that the VEGF-mediated angiogenesis in zebrafish was in part dependent on NRP1 expression. The results contributed to a better understanding of molecular mechanisms of cardiovascular development and provided a potential promoter for making inducible knockdown in zebrafish.

Origin, development and evolutionary model of shelf desertization environment in late stage of Upper Pleistocene

Based on the study of palaeo-environmental evolution in the shelves of the Eastern China Seas, the concept of ''shelf desertization'' in the late stage of Upper Pleistocene is defined; the environmental background and evolutionary process of shelf desertization are analysed. Study on the records of subbottom profiling and the data of core samples from shelf areas revealed that during low sea-level stages, the sedimentary environment in the exposed shelf plains was dominated by aeolian depositional process under cold and dry climatic conditions, i.e. under the action of strong winter-monsoon winds. Parts of the exposed marine strata were disintegrated, and aeolian sand dunes were formed on the disintegrated marine deposits, from which the finer sediment grains were blown away by wind and deposited in the downwind areas to form the derivative loess deposits. Thus a desertization environmental system was formed in the exposed shelf plains of the Eastern China Seas.

Development of an in vitro multicellular tumor spheroid model using microencapsulation and its application in anticancer drug screening and testing

In this study, an in vitro multicellular tumor spheroid model was developed using microencapsulation, and the feasibility of using the microencapsulated. multicellular tumor spheroid (MMTS) to test the effect of chemotherapeutic drugs was investigated. Human MCF-7 breast cancer cells were encapsulated in alginate-poly-L-lysine-alginate (APA) microcapsules, and a single multicellular spheroid 150 mu m in diameter was formed in the microcapsule after 5 days of cultivation. The cell morphology, proliferation, and viability of the MMTS were characterized using phase contrast microscopy, BrdU-Iabeling, MTT stain, calcein AM/ED-2 stain, and H&E stain. It demonstrated that the MMTS was viable and that the proliferating cells were mainly localized to the periphery of the cell spheroid and the apoptotic cells were in the core. The MCF-7 MMTS was treated with mitomycin C (MC) at a concentration of 0.1, 1, or 10 times that of peak plasma concentration (ppc) for up to 72 h. The cytotoxicity was demonstrated. clearly by the reduction in cell spheroid size and the decrease in cell viability. The MMTS was further used to screen the anticancer effect of chemotherapeutic drugs, treated with MC, adriamycin (ADM) and 5-fluorouracil (5-FU) at concentrations of 0.1, 1, and 10 ppc for 24, 48, and 72 h. MCF-7 monolayer culture was used as control. Similar to monolayer culture, the cell viability of MMTS was reduced after treatment with anticancer drugs. However, the inhibition rate of cell viability in MMTS was much lower than that in monolayer culture. The MMTS was more resistant to anticancer drugs than monolayer culture. The inhibition rates of cell viability were 68.1%, 45.1%, and 46.8% in MMTS and 95.1%, 86.8%, and 91.6% in monolayer culture treated with MC, ADM, and 5-FU at 10 ppc for 72 h, respectively. MC showed the strongest cytotoxicity in both MMTS and monolayer, followed by 5-FU and ADM. It demonstrated that the MMTS has the potential to be a rapid and valid in vitro model to screen chemotherapeutic drugs with a feature to mimic in vivo three-dimensional (3-D) cell growth pattern.