Lentivirus vectors construction of SiRNA targeting interference GPC3 gene and its biological effects on liver cancer cell lines Huh-7

Objective:To build GPC3 gene short hairpin interference RNA(shRNA)slow virus veclor.observe expression of Huh-7 GPC3 gene in human liver cell line proliferation apoptosis and the effect of GPC3 gene influencing on liver cancer cell growth,and provide theoretical basis for genc therapy of liver cancer.Methods:Hepatocellular carcinoma cell line Huh-7 wsa transfected by a RNA interference technique.GPC3 gene expression in a variety of liver cancer cell lines was detected by fluorescence quantitative PCR.Targeted GPC3 gene seqnences of small interfering RNA(siRNA)PGC-shRNA-GPC3 were restructured.Stable expression cell linse of siRNA were screened and established with the heplp of liposomes(lipofectamine^(TM2000))as carrier transfcetion of human liver cell lines.In order to validate siRNA interference efficiency.GPC3 siRNA mRNA expression was detected after transfection by using RT-PCR and Western blot.The absorbance value of the cells of blank group,untransfection group and transfection group,the cell cycle and cell apoptosis were calculated,and effects of GPC3 gene nn Huh-7 cell proliferation and apoptosis were observed.Results:In the liver cancer cell lines Huh-7 GPC3 gene showed high expression.PGC-shRNA-GPC3 recombinant plasmid was constructde successfully via sequencing validation.Stable recombinant plasmid transfected into liver cancer cell linse Huh-7can obviously inhibit GPC3 mRNA expression level.Conclusions:The targeted GPC3 siRNA can effectively inhibit the expression of GPC3.

Correction of Observation Interference on Z component of Geomagnetic Field From DC Power Lines

为减小直流输电线路对地磁场观测的干扰,采用数学计算的方法精确求解了线路对地磁Z分量的干扰水平。以观测点为原点建立了地磁仪器观测点坐标系,推导了观测点坐标系各坐标轴在地心坐标系中的单位矢量表达式。根据空间矢量点积的定义,得到了空间电流磁场矢量投影到地磁Z分量方向的求解公式。利用推导的公式和直流线路电流磁场三维计算模型,最终求得直流线路对地磁Z分量观测的干扰水平。由此,开发了＂直流线路对地磁观测干扰校正系统＂,并开展了地磁台站和野外联合地磁观测干扰实验。结果表明,直流线路和地磁观测台站距离不超过10km时,提出的数学模型和计算方法误差不超过5%。

Development in Researches on Reradiation Interference from UHV Power Lines

Reradiation interference(RRI) from ultra high voltage(UHV) power lines has become a hotspot for researches in electromagnetic(EM) interference between UHV power grids and adjacent radio stations.The mechanism of RRI,numerical simulations,methods of protecting distance calculation,and resonance characteristics of RRI are reviewed in this paper using results of works reported by IEEE and Chinese publications.We conclude in this review that RRI at short and medium wavelengths can be simulated using method of moment(MoM) and two commonly used models,the wire model and the surface model,which have different applicable conditions.We indicate that the accurate simulation of RRI at higher frequencies using uniform geometrical theory of diffraction is still beyond our capability because it requires studies of the relative simulation methods.We also suggest that further researches of the mechanism of RRI and the prediction of resonance frequencies above 1.7 MHz are necessary for dealing with the interference between the existing power lines and radio stations because resonance frequencies proposed by IEEE are less than 1.7 MHz.

Inhibition of Hepatitis C Virus Genotype 1a Non-Structural Proteins by Small Interference RNA in Human Hepatoma Cell Lines

Hepatitis C virus (HCV) infection and associated liver diseases are still challenging and represent a significant health care burden around the world. Although, the treatment strategies have been improved by the development of novel direct-acting antivirals, but such therapeutic options are still expensive and beyond the financial range of the most infected individuals in developing or even in resource replete countries. It demands an urgent need to search novel and improved alternate treatment strategies to treat the infection. The present study was aimed to develop an in vitro stable cell culture system, persistently expressing HCV genotype 1a non-structural genes and to characterize the inhibitory effects of synthetic siRNAs (short interference RNA) directed against the most conserved regions of nonstructural genes in an in vitro cell culture model. The continuous expression of nonstructural genes for more than 30 days post transfection was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis in stable human hepatoma cell line (Huh-7). The gene expression studies revealed significantly reduced gene expression of HCV nonstructural genes (i.e., NS2, NS4A and NS5A) both at mRNA and protein levels when treated against genome specific synthetic siRNAs in stable cell lines (51%, 47% and 54% respectively, p < 0.05). Similarly, a vivid decrease in HCV viral titer was exhibited by synthetic siRNAs in an in vitro viral replicate cell culture model (58%, 48% and 50%, respectively, p < 0.05) determined by quantitative Real-Time PCR (qPCR). Our data indicate that siRNA mediated gene silencing may be considered a promising alternate treatment strategy against HCV in combination with other effective therapeutic regimens in future.

Radio Interference Characteristic of Ultra-high Voltage AC Transmission Lines by Using Stochastic Modeling

For developing ultra-high voltage(UHV) AC power transmission systems,it is important to precisely estimate and to limit the radio interference(RI) level of power lines.Based on the stochastic characteristics in amplitude and repetition rate of induced corona current,by using the probability theory and mathematical statistics,we establish a stochastic model for the wide-sense stationary random process of corona discharges.Then combining the stochastic model with model-propagation-analysis method,the RI levels under three-phase UHV AC transmission lines are calculated.The results of the calculation based on stochastic model method and International Council on Large Electric Systems(CIGRE) excitation function are compared with that based on semi-empirical method and some other excitation functions.The stochastic model based on different excitation functions is also adopted to simulate the RI levels under finite test lines with two opened terminations.The results indicate that with the same average maximum gradient on conductor surface and the same conductor type,the number of corona discharge per unit length is one of the main reasons that causes the difference between different excitation functions.It is also concluded that for a long test line,the effect of standing wave on RI field strength is negligible in the middle of the line,but obvious near both terminations: for a 10-km line,the maximum difference in RI field strength is 2.78 dB,between the peak value of the standing wave near the ends and the steady value near the middle of the line.

Analysis of field coupling to transmission lines with random rotation over the ground

In this paper we analyze plane wave coupling to transmission lines rotating randomly over an infinite and perfectly conducting ground and present an efficient method to calculate average voltage. Under the assumption of small rotation quantity, the factors affecting the induced voltage and their effects are analyzed and then an efficient method to calculate the average voltage is presented when the distribution of the random rotation angles is uniform in [-π, π]. The results show that voltage variation is mainly due to the change of the source term. The effects of the source term increase linearly with the magnitude of the incident wave, change periodically with the rotation angle, and are larger in the high frequency range than in the low frequency range. The results show that the average voltages obtained by the proposed method agree well with those via the Monte Carlo method and the proposed method is much more efficient. The results also imply that the effect of random rotation is more important than that of random translation.

Neutrino oscillation interference phase in Kerr space time

The mass neutrino interference phases along the null trajectory and the geodesic line in Kerr space-time are studied on the plane θ = π/2. Because of the rotation object in Kerr space-time, a particle travelling along the radial geodesic must have a dragging effect produced by the angular momentum of the central object. We give the correction of the phase due to the rotation of the space-time. We find that the type-I interference phase along the geodesic remains the double of that along the null on the condition that the rotating quantity parameter a^2 is preserved and the higher order terms are negligible （e.g. a^4）. In addition, we calculate the proper oscillation length in Kerr space-time. All of our results can return to those in Schwarzschild space-time as the rotating parameter a approaches zero.

RNA interference affects tumorigenicity and expression of insulin-like growth factor-1,insulin-like growth factor-1 receptor,and basic fibroblast growth factor-2 in rat C6 glioma cells

BACKGROUND： Human gliomas are more likely to express basic fibroblast growth factor-2 （FGF-2） insulin-like growth factor-1（IGF-1）, and IGF-1 receptor （IGF-1R） than normal brain tissue. These factors activate signal transduction systems of Ras/MAPK and PI3K/Akl, which promote glioma growth. OBJECTIVE： To utilize RNA interference （RNAi） technique to down-regulate FGF-2, IGF-1, and IGF-1R gene expression, and to investigate the effects of these genes on rat C6 glioma cells, as well as the feasibility of RNAi for treating glioma. DESIGN, TIME AND SETTING： This neurooncological, randomized, controlled, in vivo and in vitro experiment, which used RNAi methodology, was performed at the Laboratory of Molecular Biology, Institute of Biochemistry, Chinese Academy of Sciences between August 2005 and February 2008. MATERIALS： Rat C6 cell lines were purchased from Shanghai Institute of Cellular Biology Affiliated to Chinese Academy of Sciences. Small interfering RNA （siRNA） was synthesized by Shanghai GenePharma. Anti-IGF-1, anti-IGF-1R, anti-FGF-2, anti-mouse and anti-rabbit IgG G1-HRP antibodies were provided by Santa Cruz Biotechnology, USA. Four to six week-old BALB/c nude mice were purchased from the Laboratory Animal Center, Chinese Academy of Sciences. METHODS： C6 glioma cells were transfected with siRNA, which was chemically synthesized in vitro to correspond to endogenous FGF-2, IGF-1, and IGF-1R genes. The inhibition ratio of targeting mRNA expression was detected by semiquantitative RT-PCR, and protein expression was determined by Western blot analysis. C6 glioma cell proliferation was observed using a growth curve C6 glioma cell apoptosis rate and cell cycle were detected by flow cytometry. C6 glioma cell growth regression was observed by transwell migration assay. In addition, nude mouse subcutaneous tumor models were used in this study. For studying the anti-tumor effects of IGF-1 and IGF-1R siRNA, two blank control groups, with six mice each, were set up： A （2.5 μg siRNA was injected one week after C6 cells were inoculated, Le., when tumor volume reached 8 mm × 8 mm） and B （siRNA was injected at the same time with C6 cells were inoculated. To study the effects of FGF-2 siRNA, the groups consisted of a blank control group, negative control group, 2.6 μg siRNA group, 4 μg siRNA group, and 5.3 μg siRNA group, with six mice each. MAIN OUTCOME MEASURES： mRNA and protein inhibition ratio of FGF-2, IGF-1, and IGF-1 R; C6 glioma cell proliferation, apoptosis, and cycle growth arrest; C6 glioma cell growth regression and subcutaneous tumorigenicity rates. RESULTS： All siRNA constructs proved to be effective. After 48 hours, transfection of 200 nmol/L siRNA resulted in a FGF-2 or IGF-1R gene inhibition ratio 〉 80% and an IGF-1 gene inhibition ratio of approximately 70%. Protein expression levels for FGF-2, IGF-1, and IGF-1R decreased in a dose-dependent manner following siRNA transfection, with an inhibition rate 〉 85%, 60%, and 50%, respectively. C6 glioma cell proliferation and apoptosis rates increased in proportion to siRNA. The apoptosis rate of C6 glioma cells induced by FGF-2, IGF-1, and IGF-1R siRNA was 39.96%, 15.07% and 22.47%, respectively （P 〈 0.01）. Transfection of 200 nmol/L IGF or IGF-1R siRNA for 48 hours suppressed C6 glioma cell migration. At 30 days after intratumoral injection of 2.6, 4, and 5.3 tJg FGF-2 siRNA, tumor growth regression rate of FGF-2 siRNA was 56%, 67%, and 86%, respectively. The tumor growth regression rate was 71.88% and 45.71%, respectively, when IGF-1 or IGF-1R siRNA was intratumorally injected 1 week after C6 glioma cell transplantation. When IGF-1 or IGF-1 R siRNA was intratumorally injected during C6 glioma cell transplantation, the tumor growth regression rate was 78.13% and 74.29%, respectively. CONCLUSION： siRNA transfection downregulated gene expression of FGF-2, IGF-1, and IGF-1R In addition, siRNA treatment markedly suppressed glioma cell proliferation, growth, and migration, and concomitantly reduced subcutaneous tumorigenicity.

A Compact Test System for Simulating Multipath Interference

Fading and inter-symbol interference (ISI) arising from multipath effects are a common source of both latency and packet errors in wireless communications. Test methods often require large environments to produce long delays that are enough to affect the communications between a transmitter and receiver. This paper presents a simple, compact test method to produce and isolate effects from multipath interference simulating these effects produced by discrete distances. Signals with controlled delays can be created and combined using multiple antennas in two isolated small (2 foot/60 cm) cubes constructed with microwave absorber. We demonstrated this with a pulsed RF signal and a signal from an 802.11 n access point with an internal antenna. This method can be further extended to provide a compact test-bed for almost any wireless interference or coexistence test.

Interference Phase of Neutrino Oscillation in Schwarzschild-de Sitter Space-Time

We study the mass neutrino interference phase in Schwarzschild-de Sitter space time along the null trajectory and the geodesic line and obtain the effects of cosmological constant A on the neutrino oscillation. Firstly, in the high energy limit, we find that the phase along the geodesic keeps the double of that along the null. Secondly, we calculate the phase on the condition that the cosmological constant, A, is a small quantity. The correction of the phase due to A is given. Finally, we calculate the proper oscillation length in Schwarzschild-de Sitter space-time, which increases because of the existence of A, compared with the result in Schwarzschild space-time. All of our results can be reduced to those in Schwarzschild space-time as A approaches to zero.

CORRECTION OF WING INTERFERENCE WITH KALMAN FILTERING IN INDUCTIVELY COUPLED PLASMA ATOMIC EMISSION SPECTROMETRY

A Kalman filter was developed for correction of wing interference in ICP-AES.Modeling wing interference theoretically instead of experimentally, the filter can compensate the shift in wavelength position in scans, and therefore reduce the effect of the interference on detection limit.

C-Fe谱线干扰修正对飞灰含碳量LIBS测量的影响

在燃煤电厂,飞灰含碳量是直接反映锅炉燃烧效率的重要指标,控制含碳量水平和低氮燃烧之间的平衡要求实现含碳量的在线(或快速)检测。将激光诱导击穿光谱技术应用于飞灰含碳量的快速测量,针对测量中248nm附近的C和Fe谱线干扰问题,提出了利用Fe谱线修正的方法以提取重叠峰中C谱线的积分强度,对比分析了Fe 248.33nm,Fe 254.60nm和Fe 272.36nm谱线分别作为Fe 247.98nm的修正谱线时提取的C修正积分强度对飞灰含碳量定标曲线和未知样品重复测量精确度的影响。研究结果表明,对C和Fe谱线干扰进行强度修正可以提高含碳量定标曲线的拟合度,并且可以显著改善低含碳量样品重复测量的精确度。但同时需要注意用于修正的Fe谱线的合理选取,防止在对低含碳量样品中C谱线强度的过度修正。从定标曲线和重复测量精确度总体评价而言,Fe 254.60nm谱线最适用于LIBS测量飞灰含碳量时的C和Fe谱线干扰的修正。

Knocking-down of Nogo-A Gene Expression in PC12 Cell Line by Plasmid-based RNAi

To study the inhibitory effect of Nogo-A shRNA on cell line PC12, the Nogo-A shRNA （short hairpin RNA, or shRNA） was designed and synthesized. The annealed shRNA template was inserted into plasmid pGenesil-1 containing enhanced green fluorescent protein （EGFP） gene by gene cloning technique to generate eukaryotic expression vector. The recombinant plasmid was transfected into PC12 cells by lipofecamine2000 and the mRNA and protein expression level of Nogo-A gene was detected by RT-PCR and Western blotting 48 h after the transfection. Gene sequencing showed that that the Nogo-A shRNA eukaryotic expression vector was successfully constructed. No significant change was found in the Nogo-A mRNA and protein expression level in empty vector-transfected group as compared with controls （P〉0.05）, while the expression level in shRNA-transfected group decreased significantly （P〈0.05）. It is concluded that the pGenesil-1/Nogo-AshRNA recombinant plasmid can effectively suppress the expression of Nogo-A gene in PC12 cells.

Statistical Analysis of Radio Interference of 1000 kV UHV AC Double-circuit Transmission Lines in Foul Weather

Analyzing the impact of radio interference(RI)variation during foul weather conditions is an area that has received limited study.This paper provides a statistical analysis of RI measurements obtained from a long-term observation station close to the world’s first commercially operating 1000 kV UHV AC double-circuit transmission line in China.During six months of observations,the impact of RI was studied on the line during fog,drizzle,and light snow and rain.It was found that RI increases linearly with the natural logarithm of the precipitation intensity.The Levenberg-Marquardt algorithm(LMA)is employed to fit the RI value with the precipitation intensity.The reasonable distribution of RI in different foul weather is verified by one-sample K-S test.This test is seen as beneficial for further RI prediction based on statistical weather mode.

Effect on proliferation and apoptosis of T24 cell lines via silencing DNMT1 with RNA interference

Expression of DNA methyltransferase 1(DNMT1),which plays an important role on aberrantly methylated CpG in the promoter regions of tumor sup-pressor genes(TSGs),is higher in bladder cancer cells than in normal bladder cells.Therefore,its overexpression is closely related to tumor formation.In this study,the eukaryotic vector pshRNA-DNMT1 was constructed and transfected into T24 cells.Levels of DNMT1 mRNA and protein were detected by reverse transcription-polymerase chain reaction(RT-PCR)and western blot.Relative to the blank control at the 24th,48th and 72nd hour after transfection of pshRNA-DNMT1,the inhibitory rates of DNMT1 mRNA levels in T24 cells were 28.44%,52.48%,70.91%,respectively.Those of DNMT1 proteins were 24.27%,57.79%,and 77.74%,respectively.Proliferation and apoptosis were assayed by MTT and flow cytometry with Annexin-V-FITC/PI staining.The growth inhibition rates of pshRNA-DNMT1 at the 24th,48th and 72nd hour after transfection of pshRNA-DNMT1 were(4.34¡0.76)%,(9.87¡1.54)%and(13.78¡1.93)%,respectively.There were statistically significant differ-ences between pshRNA-DNMT1 and the control blank at each time points(P,0.01);24,48 and 72 hours after T24 cells were transfected by pshRNA-DNMT1,the apoptosis rates of pshRNA-DNMT1 were(3.87¡0.81)%,(8.69¡1.23)%and(11.46¡1.24)%,respectively(P,0.01 vs blank control).Based on this case,our conclu-sion is that the recombinant plasmid pshRNA-DNMT1 can silence the expression of gene DNMT1 mRNA and protein effectively,and to some extent,it also can inhibit the proliferation of bladder cancer cell and promote the cellular apoptosis.

Off-line simulation of robot welding of radar pedestal

A study of the interference simulation based on robot welding of the radar pedestal was carried out by using the KUKA Sim Pro simulation software and off-line program technology. Compared with the actual robot welding process, it was found that the trajectory of the simulated robot welding process in line with that recorded in the actual welding process, and the actual limit and interference appeared at the same place as the simulation process. There was no interference phenomenon on the outside weld-seam; on the internal weld-seam, especially on the weld-joint of support plate connected to the cylinder, a phenomenon of interference appeared. It was helpful to use the simulation method to guide the actual robot welding so as to protect robot from impacting and reduce the weld defects.

gprMax干扰消除方法优化隧道壁后空洞成像特征研究

为解决传统探地雷达图像处理方法在检测隧道衬砌钢筋影响下的壁后空洞时,二维模型不能有效描述空洞三维特征,存在计算效率低、计算精度不足的问题,综合运用平均消除法、指数增益法和时间零点校正法,结合雷达信号随双程窗口时间变化的特点,引入分段函数计算步骤,提出gprMax干扰消除方法,并验证该方法在正演模拟结果处理中的可行性。结合工程实际注浆情况和隧道壁后空洞的检测过程,采用基于时域有限差分法的gprMax软件设计隧道壁后注浆空洞的三维模型,采用gprMax干扰消除方法优化图像结果,对不同工况下的正演模拟图像成像特征进行分析。结果表明:1)gprMax干扰消除方法的正演模拟处理结果与室内混凝土板中圆形空洞结构模型试验的实测结果总体一致;2)水平测线中矩形空洞信号特征呈现出“凹”字形,垂直测线中矩形空洞信号特征呈现出类似直达波的特征,由空洞的三维特征图获取数据可计算得到空洞尺寸和埋深信息;3)gprMax干扰消除方法可以有效增强埋深较大处空洞的电磁信号强度。由研究结果可知,gprMax干扰消除方法在提高了隧道壁后空洞检测数据处理效率的同时保证了计算准确性和精度。

Adaline and FIS Techniques for Fault Identification in HV Transmission Line

This paper is to identify and classify the various types of shunt and line faults in transmission line. The faults may be an insulation failure, lightning or accidental faulty operation. In a transmission line protection important factor is identifying a fault because if any error occurs in finding fault may leads to abnormal operation of the protection system. So either a disturbance or steady state variation is called power quality variation. The proposed test system is modeled based on the neural network and fuzzy algorithm. The online symmetrical components are extracted by this above algorithm. The fuzzy is used to separate the oscillating components and average components. Here input for the fuzzy is trained by using neural network. It is based on current samples and very effective in fault classifier using rule base. This method is very much suitable for online implementation.

多线并行轨道电路电容耦合干扰机理研究

高速铁路车站并行股道多,应用场景复杂,在三线并行情况下,中间股道易受两旁线路的电磁干扰影响。当2条线路均与中间股道同向时,中间股道所受电容耦合干扰为2条线路干扰之和,且邻线耦合电容越大,干扰亦越强。为减少多线并行邻线间的电容耦合干扰,可采取改变线路相对布局、采用屏蔽等设计措施,确保高速铁路车站轨道电路工作的稳定性和可靠性。