DNA Methylation Analysis of Exogenous Genes Transformed into Sheep by Different Methods

[Objective] This study aimed to investigate the methylation levels of exogenous genes and promoters and the differences of protein expression in transgenic sheep obtained by different transgenic technologies. [Method] Exogenous genes eGFP （enhanced green fluorescent protein） and FGF5 （fibroblast growth factor 5） were separately transformed into sheep by somatic cell cloning, stem cell cloning and perivitelline injection to obtain transgenic sheep, with CMV as the promoter. Bisulfite sequencing method was adopted to detect the methylation status of the promoter region and coding region of exogenous genes in tail tissues of transgenic sheep. Western blot was adopted to detect the expression level of exogenous genes. [Result] The methylation level of the promoter region with stem cell cloning was the highest, followed by somatic cell cloning, while that with perivitelline injection was the lowest; the methylation level of the eGFP coding region with perivitelline injection was the highest, followed by stem cell cloning; the methylation level of the FGF5 coding region with somatic cell cloning was higher than that with perivitelline injection. The exogenous protein expression level was negatively correlated with the methylation level of the promoter region. [Conclusion] This study indicates that different transgenic methods may influence the methylation level of exogenous genes, thus affecting exogenous gene expression.

Exact Periodic-Wave Solutions to Nizhnik-Novikov-Veselov Equation

Exact periodic-wave solutions to the generalized Nizhnik-Novikov-Veselov (NNV) equation are obtained by using the extended Jacobi elliptic-function method, and in the limit case, the solitary wave solution to NNV equation are also obtained.

Metabolically based liver damage pathophysiology in patients with urea cycle disorders-A new hypothesis

The underlying pathophysiology of liver dysfunction in urea cycle disorders(UCDs) is still largely elusive. There is some evidence that the accumulation of urea cycle(UC) intermediates are toxic for hepatocyte mitochondria. It is possible that liver injury is directly caused by the toxicity of ammonia. The rarity of UCDs, the lack of checking of iron level in these patients, superficial knowledge of UC and an underestimation of the metabolic role of fumaric acid, are the main reasons that are responsible for the incomprehension of the mechanism of liver injury in patients suffering from UCDs. Owing to our routine clinical practice to screen for iron overload in severely ill neonates, with the focus on the newborns suffering from acute liver failure, we report a case of citrullinemia with neonatal liver failure and high blood parameters of iron overload. We hypothesize that the key is in the decreased-deficient fumaric acid production in the course of UC in UCDs that causes several sequentially intertwined metabolic disturbances with final result of liver iron overload. The presented hypothesis could be easily tested by examining the patients suffering from UCDs, for liver iron overload. This could be easily performed in countries with a high population and comprehensive national register for inborn errors of metabolism. Conclusion: Providing the hypothesis is correct, neonatal liver damage in patients having UCD can be prevented by the supplementation of pregnant women with fumaric or succinic acid, prepared in the form of iron supplementation pills. After birth, liverdamage in patients having UCDs can be prevented by supplementation of these patients with zinc fumarate or zinc succinylate, as well.

Variable Separation Approach to Solve （2 ＋ 1）-Dimensional Generalized Burgers System： Solitary Wave and Jacobi Periodic Wave Excitations

By means of the standard truncated Painlevé expansion and a variable separation approach, a general variable separation solution of the generalized Burgers system is derived. In addition to the usual localized coherent soliton excitations like dromions, lumps, rings, breathers, instantons, oscillating soliton excitations, peakons, foldons, and previously revealed chaotic and fractal localized solutions, some new types of excitations — compacton and Jacobi periodic wave solutions are obtained by introducing appropriate lower dimensional piecewise smooth functions and Jacobi elliptic functions.

The Clock gene clone and its circadian rhythms in Pelteobagrus vachelli

The Clock gene,a key molecule in circadian systems,is widely distributed in the animal kingdom. We isolated a 936-bp partial c DNA sequence of the C lock gene( Pva- clock) from the darkbarbel catfish P elteobagrus vachelli that exhibited high identity with C lock genes of other species of fish and animals(65%–88%). The putative domains included a basic helix-loop-helix(b HLH) domain and two period-ARNT-single-minded(PAS) domains,which were also similar to those in other species of fish and animals. P va- Clock was primarily expressed in the brain,and was detected in all of the peripheral tissues sampled. Additionally,the pattern of P va- Clock expression over a 24-h period exhibited a circadian rhythm in the brain,liver and intestine,with the acrophase at zeitgeber time 21:35,23:00,and 23:23,respectively. Our results provide insight into the function of the molecular C lock of P. vachelli.

Invasion and Morphological Variation of the Non- Indigenous Barnacle Chthamalus challengeri(Hoek, 1883) in Yangshan Port and its Surrounding Areas

Invasive species generally possess unique characteristics that allow them to survive the invasion process in order to es- tablish and spread in new habitats. Successful invaders must resist both physical and physiological stresses associated with the changing environment. A common littoral barnacle, Chthamalus challengeri Hoek, 1883 （Crustacea, Cirripedia）, which is native to Japan, South Korea and northern China, has become established in the high-littoral zone adjacent to Yangshan Port, Shanghai, China. A comparison of the morphology of Chthamlus species from Zhoushan archipelago with previous description indicates the occur- rence of C. challengeri. The new immigrant becomes a dominant species in certain high-intertidal habitats of the adjacent area to of Yangshan Port. C. challengeri was found in part of sampling sites in Zhoushan in 2010; however, it dispersed to all the eleven sam- piing sites in 2012. Densities of C.challengeri had increased over 10 times in the last 2 years, with the highest mean value reaching 39533 ± 6243 ind. m-2 in the new habitat. The specific ratios of both operculum area （Sa） to base area （SA） and average height of pa- rietal plates （H） to length of base （L） revealed that C. challengeri displays morphological changes to resist stronger currents in the new habitats for invasion.

Effects of STI571 and p27 gene clone on proliferation and apoptosis of K562 cells

AIM: To investigate the combined effect of STI571 and p27 gene clone on the regulation of proliferation, cell cycle and apoptosis of K562 cell line.METHODS: p27 gene was obtained by RT-PCR, and its sequence was approved to be correct. Then p27-pcDNA3.1 vector was constructed and transfected into K562 cell line.p27-pcDNA3.1-K562 cell clone was screened by G418 after transfection, p27 protein was identified by Western blot.MTT was used to detect the survival rate of the cell. Flow cytometry was used to detect cell cycle and apoptosis index.RESULTS: The expression of p27 protein could be detected by Western blot in p27-pcDNA3.1-K562 cells. A strong inhibition of cell proliferation was observed in p27-pcDNA3.1-K562 cells as compared with that of the control (pcDNA3.1-K562 cells). The cells at G0/G1 phase were significantly increased, and cells at S phase were greatly declined.The apoptosis index was increased greatly after p27-pcDNA3.1-K562 cells were treated with STI571, and survival rate of the cell was markedly declined (0.35-0.58,P<0.05-0.048 vs STI571-K562 cell, 0.35-0.72, P<0.01-0.001 vs p27-K562 cell).CONCLUSION: p27 and STI571 have a synergistic action on inhibition of proliferation and induction of apoptosis on K562 cells.

TP53 Arg72Pro and CCND1 A870G Polymorphisms and Esophageal Cancer Risk

The authors examined 184 residents from Kazakhstan to reveal an association of the CCND1 gene A870G and TP53 gene Arg72Pro polymorphisms with esophageal cancer risk. 86 of them were control group and 98 were patients with esophageal cancer. DNA samples were genotyped by direct sequencing method and TaqMan allelic discrimination method. Statistical analysis was performed using GraphPad InStatTM Software and ＂Case-Control Study Estimating Calculator＂ from TAPOTILI company. A significant association was revealed between CCND1 homozygous genotype （A870A, OR=2.654） and TP53 heterozygous （Arg72Pro, OR= 1.417） and homozygous （Pro72 Pro, OR=2.860） genotypes increased risk of esophageal cancer.

Elliptic Equation and Its Direct Applications to Nonlinear Wave Equations

Elliptic equation is taken as an ansatz and applied to solve nonlinear wave equations directly. More kinds of solutions are directly obtained, such as rational solutions, solitary wave solutions, periodic wave solutions and so on.It is shown that this method is more powerful in giving more kinds of solutions, so it can be taken as a generalized method.

New Doubly Periodic Solutions for the Coupled Nonlinear Klein-Gordon Equations

By using the general solutions of a new coupled Riccati equations, a direct algebraic method is described to construct doubly periodic solutions (Jacobi elliptic function solution) for the coupled nonlinear Klein-Gordon equations.It is shown that more doubly periodic solutions and the corresponding solitary wave solutions and trigonometric function solutions can be obtained in a unified way by this method.

German Labor Sports Movement in Slovakia and Its Orientation in Period 1918-1938

This is a study of German Labor Sport organizations and unions in Slovakia in the time parameters area 1918-1938 German workers living in Slovakia started to found their own union after 1918. They got a strong support from similar organizations of German workers living in Sudety. The coordinating role was played by the German Socio-Democratic party in Slovakia. The major concentration of these unions was in Bratislava and Eastern Slovakia. Concerning their specialized field, they can be divided into tourism-practising, physical and sport ones However, the union activities were strongly influenced by political parties. Activities of every union were regional and therefore a strong mutual isolation existed

Characterization of a full-length infectious clone of bovine foamy virus 3026

The biological features of most foamy viruses(FVs) are poorly understood, including bovine foamy virus(BFV). BFV strain 3026(BFV3026) was isolated from the peripheral blood mononuclear cells of an infected cow in Zhangjiakou, China. A full-length genomic clone of BFV3026 was obtained from BFV3026-infected cells, and it exhibited more than 99% amino acid(AA) homology to another BFV strain isolated in the USA. Upon transfection into fetal canine thymus cells, the full-length BFV3026 clone produced viral structural and auxiliary proteins, typical cytopathic effects, and virus particles. These results demonstrate that the full-length BFV3026 clone is fully infectious and can be used in further BFV3026 research.

走工业服务道路 打造钢铁配供中心——走进江苏大明金属制品有限公司

2002年,江苏大明金属制品有限公司在国道312与通江大道的交汇处投资兴建无锡不锈钢市场和市内的东方钢材城、南方不锈钢市场形成三足鼎立之势。如今,东方钢材城、南方不锈钢市场仍还保持着市场经营的模式。

DIRECT CONSTRUCT OF PERIODIC SOLUTIONS TO PERTURBED KLEIN-GORDON EQUATIONS BY NEWTON ITERATION

By introducing the block estimate technique and directly using the Newton iteration method, the author constructs Cantor families of time periodic solutions to a class of nonlinear wave equations with periodic boundary conditions. The Lyapunov-Schmidt decomposition used by J. Bourgain, W. Craig and C. E. Wayne is avoided. Thus this work simplifies their framework for KAM theory for PDEs.

Almost periodic solution of Nicholson＇s blowflies model with linear harvesting term and impulsive effects

This paper is concerned with impulsive Nicholson＇s blowflies model with linear harvesting term. By using contraction mapping fixed point theorem, we obtain sufficient conditions for the existence of unique almost periodic positive solution. Moreover, we investigate exponential convergence of the almost periodic positive solution by Lyapunov functional.

TNF-α release capacity is suppressed immediately after hemorrhage andresuscitation

Purpose： It has been suggested that patients with traumatic insults are resuscitated into a state of an early systemic inflammatory response. We aimed to evaluate the influence of hemorrhagic shock and resuscitation （HSR） upon the inflammatory response capacity assessed by overall TNF-α secretion capacity of the host compared to its release from circulating leukocytes in peripheral circulation. Methods： Rats （8/group） subjected to HS （MAP of 30e35 mmHg for 90 min followed by resuscitation over 50 min） were challenged with Lipopolysaccharide （LPS）, 1 mg/kg intravenously at the end of resuscitation （HSR-LPS group） or 24 h later （HSR-LPS24 group）. Control animals were injected with LPS without bleeding （LPS group）. Plasma TNF-α was measured at 90 min after the LPS challenge. In addition, whole blood （WB） was obtained either from healthy controls （CON） immediately after resuscitation （HSR）, or at 24 h post-shock （HSR 24）. WB was incubated with LPS （100 ng/mL） for 2 h at 37 C. TNF-α concentration and LPS binding capacity （LBC） was determined. Results： Compared to LPS group, HSR followed by LPS challenge resulted in suppression of plasma TNF-a in HSR-LPS and HSR-LPS24 groups （1835 ± 478, 273 ± 77, 498 ± 200 pg/mL, respectively）. Compared to CON the LPS-induced TNF-a release capacity of circulating leukocytes ex vivo was strongly declined both at the end of resuscitation （HSR） and 24 h later （HSR24） （1012 ± 259, 313 ± 154, 177 ± 63 ng TNF/mL, respectively）. The LBC in WB was similar between CON and HSR and only moderately enhanced in HSR24 （57 ± 6, 56 ± 6, 71 ± 5 %, respectively）. Conclusion： Our data suggest that the overall inflammatory response capacity is decreased immediately after HSR, persisting up to 24 h, and is independent of LBC.

Time Periodic Electroosmotic Flow of The Generalized Maxwell Fluids in a Semicircular Microchannel

Analytical solutions are presented using method of separation of variables for the time periodic electroosmotic flow (EOF) of linear viscoelastic fluids in semicircular microchannel. The linear viscoelastic fluids used here are described by the general Maxwell model. The solution involves analytically solving the linearized Poisson-Boltzmann (P -B) equation, together with the Cauchy momentum equation and the general Maxwell constitutive equation. By numerical computations, the influences of electric oscillating Reynolds number Re and Deborah number De on velocity amplitude are presented. For small Re, results show that the larger velocity amplitude is confined to the region near the charged wall when De is small. With the increase of the Deborah number De, the velocity far away the charged wall becomes larger for large Deborah number De. However, for larger Re, the oscillating characteristic of the velocity amplitude occurs and becomes significant with the increase of De, especially for larger Deborah number.

Comparing gene expression profiles of Kashin-Beck and Keshan diseases occurring within the same endemic areas of China

In this study, differentially expressed genes in peripheral blood from patients with Kashin-Beck disease and Keshan disease were compared to further investigate the etiology and pathogenesis of both diseases, which occur in a common endemic area of China. Twenty Kashin-Beck disease patients and 12 healthy controls, and 16 Keshan disease patients and 16 healthy controls, were grouped into four pairs. Patients and controls were selected from common endemic areas for the two diseases. Total RNA was isolated from peripheral blood mononuclear cells from all patients and controls, and gene expression profiles analyzed by oligonucleotide microarrays. Sixteen genes differentially expressed in both Kashin-Beck disease and Keshan disease (versus controls) were identified, and comprised nine genes showing synchronous and seven asynchronous expression. The Comparative Toxicogenomics Database shows that expression and biological function of these genes can be affected by multiple environmental factors, including mycotoxin and selenium content, potential environmental risk factors for the two diseases. Thus, these shared differentially expressed genes may contribute to the distinct organ lesions, caused by common environmental risk factors of Kashin-Beck disease and Keshan disease.