Alignment Prediction of Air Spring Vibration Isolation System for Marine Shafting Based on Genetic Algorithm-Back Propagation Neural Network.

Shafting alignment plays an important role in the marine propulsion system, which affects the safety and stability of ship operation. Air spring vibration isolation systems (ASVISs) for marine shafting can not only reduce mechanical noise but also help control alignment state by actively adjusting air spring pressures. Alignment prediction is the first and a key step in the alignment control of ASVISs. However, in large-scale ASVISs, due to factors such as strong interference and raft deformation, alignment prediction faces problems such as alignment measurement sensors failure and difficulty in establishing a mathematical model. To address this problem, a data model for predicting alignment state is developed based on a back propagation (BP) neural network, fully taking advantage of its self-learning and self-adaption abilities. The proposed model exploits the collected data in the ASVIS instead of the alignment measurement data to calculate the alignment state, providing another alignment prediction approach. Then, in order to solve the local optimum issue of BP neural network, we introduce the genetic algorithm (GA) to optimize the weights and thresholds of the BP neural network, and an improved GA-BP model is designed. The GA-BP model can leverage the advantages of the global search capability of GA as well as the BP neural network's fast convergence in local search. Finally, we conduct experiments on a real ASVIS and evaluate the prediction models using different criteria. The experimental results show that the proposed prediction model with the GA-BP neural network can accurately predict the alignment state, with a mean-square error (MSE) of 0.0114. And compared to the BP neural network, the GA-BP neural network reduces the MSE by approximately 74%.

Mechanical characteristics analysis of high dimensional vibration isolation systems based on high-static-low-dynamic stiffness technology.

Large floating raft vibration isolation systems (FRVISs) based on high-static-low-dynamic stiffness (HSLDS) technology offer excellent low frequency vibration isolation performance with broad application prospects. However, the design process for these complex high-dimensional coupled nonlinear systems remains poorly developed, particularly when applied for ocean-going vessels that experience rolling and pitching motions. The present work addresses this issue by establishing a six-degree-of-freedom HSLDS vibration isolation model for FRVISs composed of eight isolators, and the model is applied to fully analyze the swing stability and multidimensional vibration isolation performance of these systems. The influence of nonlinearity on the mechanical properties of the vibration isolators is analyzed more clearly by assuming that each vibration isolator realizes nonlinear HSLDS characteristics in the z direction and linear characteristics in the x and y directions. The results demonstrate that the swing displacement responses of the system are greatly reduced under weak nonlinearity, which reflects the high static stiffness and high static stability characteristics of an HSLDS system. The multidimensional vibration isolation performance of the system is evaluated according to the impacts of nonlinearity, the installation height Hz of the isolators, and the relative position Dr of the two middle isolators. The results of analysis demonstrate that applying a value of Hz = 0 produces the best vibration isolation performance overall under strong nonlinearity by avoiding unnecessary secondary peaks in the force transmission rate under harmonic mechanical excitation and ensuring a maximum high-frequency vibration isolation effect. However, applying a weak nonlinearity is better than a strong nonlinearity if Hz is not zero. In contrast, Dr has little effect on the vibration isolation effect of the raft in the x, y, and z directions. Therefore, an equidistant installation with Dr = 0.5 would be considered ideal from the standpoint of installation stability.

Numerically Exploring the Potential of Abating the Energy Flow Peaks through Tough, Single Network Hydrogel Vibration Isolators with Chemical and Physical Cross-Links.

Traditional vibration isolation systems, using natural rubber vibration isolators, display large peaks for the energy flow from the machine source and into the receiving foundation, at the unavoidable rigid body resonance frequencies. However, tough, doubly cross-linked, single polymer network hydrogels, with both chemical and physical cross-links, show a high loss factor over a specific frequency range, due to the intensive adhesion-deadhesion activities of the physical cross-links. In this study, vibration isolators, made of this tough hydrogel, are theoretically applied in a realistic vibration isolation system, displaying several rigid body resonances and various energy flow transmission paths. A simulation model is developed, that includes a suitable stress-strain model, and shows a significant reduction of the energy flow peaks. In particular, the reduction is more than 30 times, as compared to the corresponding results using the natural rubber. Finally, it is shown that a significant reduction is possible, also without any optimization of the frequency for the maximum physical loss modulus. This is a clear advantage for polyvinyl alcohol hydrogels, that are somewhat missing the possibility to alter the frequency for the maximum physical loss, due to the physical cross-link system involved-namely, that of the borate esterification.

[Establishment and optimization of systems for protoplasts isolation of soybean and chickpea that used in subcellular location].

Young leaves of Kabuli chickpea as well as soybean Xiangdou No.3, which are the current plants that studied in our laboratory were selected as materials. Effects on protoplasts yield and survival rate of different enzyme combination, concentration of D-Mannitol in enzyme combinations, pH of enzyme combinations and enzymolysis time are detected. The results showed that, the best condition for Xiangdou No.3 leaf protoplasts isolation is to rotate the cut materials for 6 hours in enyzme solution under temperature of 27 ℃ and rotate speed of 45 r/min for 6 h. Onozuka R-10 (0.5%), Hemicellulase (0.8%), Macerozyme R-10 (0.8%) in combination with Pectolyase Y-23 (0.4%) dissolving in CPW solution with MES (0.1%) and Mannitol (10%), pH 6.0 was found best for protoplasts isolation of Xiangdou No.3 leaves.The best condition for protoplasts isolation of Kabuli chickpea is to put the cut materials into enzymatic hydrolysate enzymolyse for 7 to 8 hours under temperature of 27 ℃ and rotate speed of 45 r/min on water bath shaker, the optimum combination of enzyme consists of Onozuka R-10 (0.5%), Hemicellulase (0.8%), Macerozyme R-10 (0.8%), MES (0.1%) and Mannitol (10%) dissolved in CPW solution with pH 4.8. The protoplasts prepared with the methods above are used in subcellular location and the effects show well.

High-Throughput Analysis of Mammalian Receptor Tyrosine Kinase Activation in Yeast Cells.

Tyrosine phosphorylation is an essential posttranslational modification in intracellular signaling molecules. Since tyrosine phosphorylation occurs in less than 0.1 % of all phosphorylated amino acids in mammalian cells, it is difficult to detect the nascent phosphotyrosine at a high signal-to-noise ratio due to high intracellular backgrounds (i.e., unexpected crosstalks among endogenous signaling molecules). In order to address this issue, we reconstituted the mammalian signaling pathway involving an extracellular ligand and a receptor tyrosine kinase (RTK) in Saccharomyces cerevisiae, a lower eukaryote that lacks endogenous tyrosine kinases. In this chapter, we describe a method for high-throughput analysis of ligand-receptor interaction by combining the yeast cell-surface display technique with an automated single-cell analysis and isolation system. Yeast cells coexpressing the cell-wall-anchored form of the human epidermal growth factor (EGF) and the human EGF receptor (EGFR) fused with a signal peptide at the N terminus facilitated the interaction of EGF with EGFR in an autocrine manner, followed by EGFR oligomerization and subsequent autophosphorylation. Furthermore, yeast cells expressing cell-wall-anchored forms of a conformationally constrained random peptide library instead of EGF are treated with a fluorophore-labeled anti-phosphorylated EGFR antibody and then subjected to the automated single-cell analysis and isolation system. The yeast cells with the highest level of fluorescence were shown to display novel and efficient EGFR agonistic peptides. Thus, our yeast display technique serves as a quantitative measurement for RTK activation, which is applicable to high-throughput de novo screening of RTK agonistic peptides.

Establishment and optimization of systems for protoplasts isolation of soybean and chickpea that used in subcellular location / 生物工程学报

Young leaves of Kabuli chickpea as well as soybean Xiangdou No.3, which are the current plants that studied in our laboratory were selected as materials. Effects on protoplasts yield and survival rate of different enzyme combination, concentration of D-Mannitol in enzyme combinations, pH of enzyme combinations and enzymolysis time are detected. The results showed that, the best condition for Xiangdou No.3 leaf protoplasts isolation is to rotate the cut materials for 6 hours in enyzme solution under temperature of 27 ℃ and rotate speed of 45 r/min for 6 h. Onozuka R-10 (0.5%), Hemicellulase (0.8%), Macerozyme R-10 (0.8%) in combination with Pectolyase Y-23 (0.4%) dissolving in CPW solution with MES (0.1%) and Mannitol (10%), pH 6.0 was found best for protoplasts isolation of Xiangdou No.3 leaves.The best condition for protoplasts isolation of Kabuli chickpea is to put the cut materials into enzymatic hydrolysate enzymolyse for 7 to 8 hours under temperature of 27 ℃ and rotate speed of 45 r/min on water bath shaker, the optimum combination of enzyme consists of Onozuka R-10 (0.5%), Hemicellulase (0.8%), Macerozyme R-10 (0.8%), MES (0.1%) and Mannitol (10%) dissolved in CPW solution with pH 4.8. The protoplasts prepared with the methods above are used in subcellular location and the effects show well.

Comparison of RNA isolation methods from insect larvae.

Isolating RNA from insects is becoming increasingly important in molecular entomology. Four methods including three commercial kits RNeasy Mini Kit (Qiagen), SV Total RNA isolation system (Promega), TRIzol reagent (Invitrogen), and a cetyl trimethylammonium bromide (CTAB)-based method were compared regarding their ability to isolate RNA from whole-body larvae of Thaumatotibia leucotreta (Meyrick), Thanatophilus micans (F.), Plutella xylostella (L.), and Tenebrio molitor (L.). A difference was observed among the four methods regarding RNA quality but not quantity. However, RNA quality and quantity obtained was not dependent on the insect species. The CTAB-based method produced low-quality RNA and the Trizol reagent produced partially degraded RNA, whereas the RNeasy Mini Kit and SV Total RNA isolation system produced RNA of consistently high quality. However, after reverse transcription to cDNA, RNA produced using all four extraction methods could be used to successfully amplify a 708 bp fragment of the cytochrome oxidase I gene. Of the four methods, the SV Total RNA isolation system showed the least amount of DNA contamination with the highest RNA integrity number and is thus recommended for stringent applications where high-quality RNA is required. This is the first comparison of RNA isolation methods among different insect species and the first to compare RNA isolation methods in insects in the last 20 years.

Single-cell-based breeding: Rational strategy for the establishment of cell lines from a single cell with the most favorable properties.

For efficient biomolecule production (e.g., antibodies, recombinant proteins), mammalian cells with high expression rates should be selected from cell libraries, propagated while maintaining a homogenous expression rate, and subsequently stabilized at their high expression rate. Clusters of isogenic cells (i.e., colonies) have been used for these processes. However, cellular heterogeneity makes it difficult to obtain cell lines with the highest expression rates by using single-colony-based breeding. Furthermore, even among the single cells in an isogenic cell population, the desired cell properties fluctuate stochastically during long-term culture. Therefore, although the molecular mechanisms underlying stochastic fluctuation are poorly understood, it is necessary to establish excellent cell lines in order to breed single cells to have higher expression, higher stability, and higher homogeneity while suppressing stochastic fluctuation (i.e., single-cell-based breeding). In this review, we describe various methods for manipulating single cells and facilitating single-cell analysis in order to better understand stochastic fluctuation. We demonstrated that single-cell-based breeding is practical and promising by using a high-throughput automated system to analyze and manipulate single cells.