Constitutive expression of thymidylate synthase from LCDV-C induces a transformed phenotype in fish cells

Thymidylate synthase (TS), an essential enzyme in DNA synthesis and repair, plays a key role in the events of cell cycle regulation and tumor formation. Here, an investigation was presented about subcellular location and biological function of viral TS from lymphocystis disease virus from China (LCDV-C) in fish cells. Fluorescence microscopy revealed that LCDV-C TS was predominantly localized in the cytoplasm in fish cells. Cell cycle analysis demonstrated that LCDV-C TS promoted cell cycle progression into S and G2/M phase in the constitutive expressed cells. As a result, the cells have a faster growth rate compared with the control cells as revealed by cell growth curves. For foci assay, the TS-expressed cells gave rise to foci 4-5 weeks after incubation. Microscopic examination of the TS-induced foci revealed multilayered growth and crisscross morphology characteristic of transformed cells. Moreover, LCDV-C TS predisposed the transfected cells to acquire an anchorage-independent phenotype and could grow in 0.3% soft agar. So the data reveal LCDV-C TS is sufficient to induce a transformed phenotype in fish cells in vitro and exhibits its potential ability in cell transformation. To our knowledge, it is the first report on viral TS sequences associated with transforming activity. (C) 2007 Elsevier Inc. All rights reserved.

Subcellular localization and characterization of G protein-coupled receptor homolog from lymphocystis disease virus isolated in China

G protein-coupled receptors (GPCRs) constitute a large superfamily involved in various types of signal transduction pathways, and play an important role in coordinating the activation and migration of leukocytes to sites of infection and inflammation. Viral GPCRs, on the other hand, can help the virus to escape from host immune surveillance and contribute to viral pathogenesis. Lymphocystis disease virus isolated in China (LCDV-C) contains a putative homolog of cellular GPCRs, LCDV-C GPCR. In this paper, LCDV-C GPCR was cloned, and the subcellular localization and characterization of GPCR protein were investigated in fish cells. LCDV-C GPCR encoded a 325-amino acid peptide, containing a typical seven-transmembrane domain characteristic of the chemokine receptors and a conserved DRY motif that is usually essential for receptor activation. Transient transfection of GPCR-EGFP in fathead minnow (FHM) cells and epithelioma papulosum cyprini (EPC) cells indicated that LCDV-C GPCR was expressed abundantly in both the cytoplasm and nucleoplasm. Transient overexpression of GPCR in these two cells cannot induce obvious apoptosis. FHM cells stably expressing GPCR showed enhanced cell proliferation and significant anchorage-independent growth. The effects of GPCR protein on external apoptotic stimuli were examined. Few apoptotic bodies were observed in cells expressing GPCR treated with actinomycin D (ActD). Quantitative analysis of apoptotic cells indicated that a considerable decrease in the apoptotic fraction of cells expressing GPCR, compared with. the control cells, was detected after exposure to ActD and cycloheximide. These data suggest that LCDV-C GPCR may inhibit apoptosis as part of its potential mechanism in mediating cellular transformation.

悬索桥隧道式锚碇与围岩系统的破坏模式研究

The anchorages are unparalleled structures only in a suspension bridge, and as main bearing facilities, play an important role in connecting the superstructures and the ground. The tunnel anchorage, as one alternative type of the anchorages, has more advantages over its counterpart, the gravity anchorage. With the tunnel anchorages adopted, not only can surface excavation be reduced to protect the environment, and natural condition of the rock be utilized and potential bearing capacity of surrounding rock be mobilized to save engineering cost, but also the technological predominance of auxiliary engineering measures, such as prestressed concrete, anchoring piles, rock anchors and collar beam between the two separated anchorages, can be easily cooperated to work together harmoniously under the circumstances of poor rock quality. There are plentiful high mountains and deep canyons in west part of China, and long-span bridge construction is inevitably encountered in order to realize leapfrogging development of the transportation infrastructure. Western mountainous areas usually possess the conditions for constructing tunnel anchorages, and therefore, the tunnel anchorages, which are conformed to the conception of resource conservative and sustainable society, extremely have application and popularization value in western underdeveloped region. The scientific and technological problem about the design, construction and operation of tunnel anchorages should be further investigated. Combining the engineering of western tunnel anchorages for the Balinghe Suspension Bridge, this paper probed into the survey method and in-situ test method for tunnel anchorages, scientific rock quality evaluation of surrounding rock to provide reasonable physical and mechanical parameters for design, construction and operation of tunnel anchorages, bearing capacity estimation for tunnel anchorage, deformation prediction of the anchorage-rockmass system, tunnel-anchorage slope stability analysis and the evaluation of excavation stability and degree of safety of the anchorage tunnel. The following outcomes were obtained: 1. Materials of tunnel anchorages of suspension bridge built (and in progress) at home and abroad were systematically sorted out, with the engineering geological condition and geomechanical property of surrounding rock around the anchorage tunnel, the design size of anchorages and the construction method of anchorage tunnel paid more emphasis on, to unveil the internal relationship between the engineering geological conditions of surrounding rock and the design size and axis angle of anchorages and provide references for future design, construction and study of tunnel anchorages. 2. Physical and mechanical parameters were recommended based on three domestic and foreign methods of rock quality evaluation. 3. In-situ tests, adopting the back-thrust method, of two kinds of reduced scale model, 1/30 and 1/20, for the tunnel anchorages were conducted in the declining exploration drift with rock mass at the test depth being the same as surrounding rock around real anchorages, and reliable field rockmass displacement data were acquired. Attenuation relation between the increment of distance from the anchorage and the decrement of rockmass displacement under maximum test load, and influential scope suffered by anchorage load were obtained. 4. Using similarity theory, the magnitude of real anchorage and rockmass displacement under design load and degree of safety of the anchorage system were deduced. Furthermore, inversion analysis to deformation modulus of slightly weathered dolomite rock, the surrounding rock of anchorage tunnel, was performed by the means of numerical simulation. 5. The influential law of the geometrical size to the limit bearing capacity of tunnel anchorage was studied. 6. Based on engineering geological survey data, accounting for the combination of strata layer and adverse discontinuities, the failure patterns of tunnel anchorage slope were divided into three modes: sliding of splay saddle pier slope, superficial-layer slippage, and deep-layer slippage. Using virtual work principle and taking anchorage load in account, the stability of the three kinds of failure patterns were analyzed in detail. 7. The step-by-step excavation of anchorage tunnel, the numerical overload and the staged decrement of rock strength parameters were numerically simulated to evaluate the excavation stability of surrounding rock around anchorage tunnel, the overload performance of tunnel anchorage, and the safety margin of strength parameters of the surrounding rock.

边坡工程的极限分析上限法

Theory of limit analysis include upper bound theorem and lower bound theorem. To deal with slope stability analysis by limit analysis is to approximate the real solution from upper limit and lower limit. The most used method of limit analysis is upper bound theorem, therefore it is often applied to slope engineering in many cases. Although upper bound approach of limit analysis can keep away from vague constitutive relation and complex stress analyses, it also can obtain rigorous result. Assuming the critical surface is circular slip surface, two kinematically admissible velocity fields for perpendicular slice method and radial slice method can be established according to the limit analysis of upper bound theorem. By means of virtual work rate equation and strength reduction method, the upper-bound solution of limit analysis for homogeneous soil slope can be obtained. A log-spiral rotational failure mechanism for homogeneous slope is discussed from two different conditions which represent the position of shear crack passing the toe and below the toe. In the dissertition, the author also establishes a rotational failure mechanics with combination of different logarithmic spiral arcs. Furthermore, the calculation formula of upper bound solution for inhomogeneous soil slope stability problem can be deduced based on the upper bound approach of rigid elements. Through calculating the external work rate caused by soil nail, anti-slide pile, geotechnological grid and retaining wall, the upper bound solution of safety factor of soil nail structure slope, slip resistance of anti-slide pile, critical height of reinforced soil slope and active earth pressure of retaining wall can be obtained by upper bound limit analysis method. Taking accumulated body slope as subject investigated, with study on the limit analysis method to calculate slope safety factor, the kinematically admissible velocity fields of perpendicular slice method for slope with broken slip surface is proposed. Through calculating not only the energy dissipation rate produced in the broken slip surfaces and the vertical velocity discontinuity, but also the work rate produced by self-weight and external load, the upper bound solution of slope with broken slip surface is deduced. As a case study, the slope stability of the Sanmashan landslide in the area of the Three Gorges reservoir is analyzed. Based on the theory of limit analysis, the upper bound solution for rock slope with planar failure surface is obtained. By means of virtual work-rate equation, energy dissipation caused by dislocation of thin-layer and terrane can be calculated; furthermore, the formulas of safety factor for upper bound approach of limit analysis can be deduced. In the end, a new computational model of stability analysis for anchored rock slope is presented after taking into consideration the supporting effect of rock-bolts, the action of seismic force and fissure water pressure. By using the model, not only the external woke-rate done by self-weight, seismic force, fissure water pressure and anchorage force but also the internal energy dissipation produced in the slip surface and structural planes can be totally calculated. According to the condition of virtual work rate equation in limit state, the formula of safety factor for upper bound limit analysis can be deduced.

预应力结构抗滑桩设计计算方法及其在滑坡治理中的应用研究

Landslide is a kind of serious geological hazards and its damage is very great. In recent years, landslides become more and more frequent along with increase of scale of engineering constructions and cause greater loss. Consequently, how to protect landslides has become important research subject in the engineering field. This paper improves the method how to compute landslide thrust and solves the irrational problem in the design of piles because of the irrational landslide thrust according to the theory and technology of existed anti-slide piles and pre-stressed cable anti-slide piles. Modern pre-stressing technology has been introduced and load balancing method has been used to improve the stressing behavior of anti-slide piles. Anchor cables, anti-slide piles and modern pre-stressing technology have been used to prevention complicated landslide. It is an important base to select values for the landslide thrust. An improved method to calculate design thrust of anti-slide piles has been presented in this paper on the base of residual thrust method by comparing existing methods to select values of landslide thrust in the design of anti-slide piles. In the method, residual landslide thrust behind the anti-slide piles and residual skid resistance before the piles has been analyzed, equitable distribution of residual landslide thrust behind the piles has been realized, and the method to select value of design thrust becomes more reasonable. The pre-stressed cable anti-slide piles are developed from the common anti-slide piles and are common method to prevent landslide. Their principle is that internal force of anti-slide piles is adjusted and size of section is diminished by changing constraint conditions of anti-slide piles. For landslides with deep slip surface and large scale of slopes, limitation of the method appears. Such landslides are in need of long piles and anchor cables which are not only non-economic but also can generate larger deformation and leave potential danger after prevention. For solving the problem, a new kind of anti-slide piles, inner pre-stressing force anti-slide piles, is presented in this paper, and its principle is that an additional force, which is generated in the inner anti-slide piles by arranging pre-stressed reinforcement or tight wire in a certain form in interior of anti-slide piles and stretching the steel reinforcement or tight wire, may balance out the internal force induced by landslide thrust whole or partly (load balancing method). The method will change bending moment which anti-slide piles are not good at bearing into compressive stress which piles are good at bearing, improve stressing performance of anti-slide piles greatly, diminish size of section, and make anti-slide piles not fissured in the natural service or postpone appearance of the fissures, and improve viability of anti-slide piles. Pre-stressed cable anti-slide piles and inner pre-stressing force anti-slide piles go by the general name of pre-stressed structure anti-slide piles in the paper, and their design and calculation method is also analyzed. A new calculation method is provided in the paper for design of anti-slide piles. For pre-stressed structure anti-slide piles, a new computation mode is firstly presented in the paper on the foundation of cantilever piles. In the mode, constraint form of load-bearing section of the anti-slide piles should be confirmed according to reservoir conditions in order to figure out amount of pre-stress of the anchor cables, and internal force should be analyzed for the load-bearing section of pre-stressed structure anti-slide piles so as to confirm anchorage section of anti-slide piles. Pre-stressed cables of the pre-stressed cable anti-slide piles can be arranged as required. This paper analyzes the load-bearing section of single-row and double-row pre-stressed cable anti-slide piles and provides a calculation method for design of the pre-stressed cable anti-slide piles. Inner pre-stressing force anti-slide piles are a new kind of structural style. Their load-bearing section is divided into four computation modes according to whether pre-stressed cables are applied for exterior of the anti-slide piles, and whether single-row or double-row exterior pre-stressed cables are applied. The load balancing method is used to analyze the computation modes for providing a method to design the inner pre-stressing force anti-slide piles rationally. Pre-stressed cable anti-slide piles and inner pre-stressing force anti-slide piles are applied to research on Mahe landfall in Yalong Lenggu hydropower station by the improved method to select value of design thrust of anti-slide piles. A good effect is obtained in the analysis.