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PERMANENCE AND ASYMPTOTIC PROPERTIES OF NONLINEAR DELAY DIFFERENCE EQUATIONS
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作者 李万同 《Applied Mathematics and Mechanics(English Edition)》 SCIE EI 2003年第11期1273-1280,共8页
The asymptotic behavior of a class of nonlinear delay difference equation wax studied . Some sufficient conditions are obtained for permanence and global attractivity . The results can be applied to a claxs of nonline... The asymptotic behavior of a class of nonlinear delay difference equation wax studied . Some sufficient conditions are obtained for permanence and global attractivity . The results can be applied to a claxs of nonlinear delay difference equations and to the delay discrete Logistic model and some known results are included. 展开更多
关键词 global attractivity higher-order nonlinear difference equation PERMANENCE DELAY
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IN VIVO COMPARATIVE OBSERVATION ON THE INVASIVENESS OF VARIOUS ORGANS BY DIFFERENT LEUKEMIA CELLS
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作者 褚建新 应红光 丁立 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 1990年第2期27-30,共4页
Using patho-morphological method and transplantation bio-assay, the in vivo invasiveness of leukemia cells is three transplantable mouse T cell leukemia models was comparatively studied. The results showed that the in... Using patho-morphological method and transplantation bio-assay, the in vivo invasiveness of leukemia cells is three transplantable mouse T cell leukemia models was comparatively studied. The results showed that the invasion to the liver was consistent, but that to other organs was obviously different. L615 and L7212 leukemia cells preferred to the bone marrow and spleen than to the peritoneum while L7811 leukemia cells were just the opposite. Transplantation bio-assay demonstrated that leukemia cells were present in the bone marrow of L615 mice as early as 6 hours after leukemic cell inoculation, but no leukemia cells was detected in bone marrow of L7811 mice 2 days after inoculation. In the terminal phase, L615 mice bone marrow became filled with leukemia cells, but L7811 mice bone marrow contained only a few leukemia cells. The difference of invasiveness of leukemia cells among organs is probably related to "homing" receptor. The same type of leukemia cells may possess multiple "homing" receptor. 展开更多
关键词 IN VIVO COMPARATIVE OBSERVATION ON THE INVASIVENESS OF VARIOUS ORGANS BY DIFFERENT LEUKEMIA CELLS bone
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DIFFERENCES IN EXPRESSION AND REGULATION BETWEEN TRANSFORMED CELLS OF THE HUMAN GASTRIC CARCINOMA ONCOGENE Ha-ras AND THE UNTRANSFORMED PARENT CELLS
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作者 韩复生 刘淑萍 +5 位作者 宋建国 袁艳华 张维 施华 邓国仁 刘培楠 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 1989年第1期20-25,共6页
An Ha-ras oncogene was isolated from a cell line of gastric carcinoma called BGE-823 in order to elucidate genetic control and the influence of DNA sequences. The oncogene was cloned and identified as a single nucleot... An Ha-ras oncogene was isolated from a cell line of gastric carcinoma called BGE-823 in order to elucidate genetic control and the influence of DNA sequences. The oncogene was cloned and identified as a single nucleotide substitution of thymine for guanine in the 12th codon through the sequencing of its first axon. We compared the differences of expression and regulation between the transformed Ha-ras cells and untransformed parent cells. Data indicated that the expression of Ha-ras in the transformed cells was five-fold higher than in the untransformed cells and that the Ha-ras gene in the former was hypersensitive toward DNase I. In addition, a nuclear protein of 35 kilodaltons bound strongly to the 2.5 Kb fragment located upstream of the 6.6 Kb Ha-ras gene and contained a CC rich region. These results suggest that there might be another mechanism of activation for the ras gene besides point mutation. 展开更多
关键词 gene DIFFERENCES IN EXPRESSION AND REGULATION BETWEEN TRANSFORMED CELLS OF THE HUMAN GASTRIC CARCINOMA ONCOGENE Ha-ras AND THE UNTRANSFORMED PARENT CELLS
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Image Segmentation Based on Period Difference of the Oscillation
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作者 王直杰 张珏 +1 位作者 范宏 柯克峰 《Journal of Donghua University(English Edition)》 EI CAS 2004年第1期68-71,共4页
A new method for image segmentation based on pulse neural network is proposed. Every neuron in the network represents one pixel in the image and the network is locally connected. Each group of the neurons that corresp... A new method for image segmentation based on pulse neural network is proposed. Every neuron in the network represents one pixel in the image and the network is locally connected. Each group of the neurons that correspond to each object synchronizes while different groups of the neurons oscillate at different period. Applying this period difference, different objects are divided. In addition to simulation, an analysis of the mechanism of the method is presented in this paper. 展开更多
关键词 Image segmentation neural network SYNCHRONIZATION
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Determination of Teicoplanin A2's Partition Coefficient in Different Liquid-Liquid Extraction Systems 被引量:2
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作者 陈勇 Bin Mat Hanapi +2 位作者 徐志南 金志华 岑沛霖 《Chinese Journal of Chemical Engineering》 SCIE EI CAS CSCD 2004年第4期493-497,共5页
Teicoplanin is one of the macrocyclic glycopeptide antibiotics, which is active against Gram-positive bacteria, and has attracted a lot of attention in the field of chiral separation recently. In this work, the partit... Teicoplanin is one of the macrocyclic glycopeptide antibiotics, which is active against Gram-positive bacteria, and has attracted a lot of attention in the field of chiral separation recently. In this work, the partition coefficients and extraction ratio of teicoplanin in three different solvent systems were studied: conventional extraction, reactive extraction and reverse micelle extraction. With conventional solvent extraction, n-butanol demonstrated high partition coefficient for teicoplanin, but low extraction ratio because of its high solubility in water. Reactive extraction of teicoplanin showed the highest partition coefficient with almost 100% recovery in organic phase when tri-n-octylmethyl ammonium chloride (TOMAC) was used as extractant and pH value was above 5.0. A reverse micelle system, consisted of isooctane +10mmol-L-1 TOMAC +1% n-octanol, also offered high separation factor for teicoplanin. The results are beneficial for the design of teicoplanin separation and purification process. 展开更多
关键词 TEICOPLANIN partition coefficients conventional extraction reverse micelle extraction
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Measurement of the Average Molecular Weight of Cotton Cellulose Crosslinked by a Polycarboxylic Acid at Different pH Using Multiple Angle Light Scattering Photometer in a DMAc/LiCl Solvent System
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作者 毛志平 YANG Charles Q 《Journal of Donghua University(English Edition)》 EI CAS 2003年第4期5-7,共3页
Durable press finishing of cotton fabrics with polycarboxylic acid increases fabric wrinkle-resistance at the expense of its mechanical strength. Severe tensile strength loss is the major disadvantage for wrinkle resi... Durable press finishing of cotton fabrics with polycarboxylic acid increases fabric wrinkle-resistance at the expense of its mechanical strength. Severe tensile strength loss is the major disadvantage for wrinkle resistant cotton fabrics.Tensile strength loss of cotton fabric crosslinked by a polycarboxylic acid can be attributed to depolymerization and crosslink of cellulose molecules. Measurement of the molecular weight of cotton fabric before and after crosslinked by polycarboxylic acids can offer a possibility of direct understanding of the depolymerization. In this research, amultiple angle laser light scattering photometer was used todetermine the absolute molecular weight of cotton fabric treated with BTCA at different pH and then hydrolyzed with 0. 5 M NaOH solution at 50℃ for 144 h. The results indicate that average molecular weights of cotton fabric treated with polycarboxylic acids at different pH are almost the same. 展开更多
关键词 cotton cellulose DEPOLYMERIZATION laser light scattering photometer molecular weight
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Study on the Similarities and Differences of Body Measurement Terminology between ASTM and China GB Standard
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作者 方方 张渭源 张文斌 《Journal of Donghua University(English Edition)》 EI CAS 2003年第4期41-48,共8页
The similarities and differences of ASTM and China GB standard are studied in three aspects:measure instrument,terminology and applicable field.They are similar on the measuring apparatus and GB has less measurements,... The similarities and differences of ASTM and China GB standard are studied in three aspects:measure instrument,terminology and applicable field.They are similar on the measuring apparatus and GB has less measurements,such as girth,length and width measurements than ASTM and it lack across chest width,back width,total crotch length and shoulder slope which are important measurements in pattern making.ASTM classifies its standards according to the customers' size,gender and age.So we think GB standard could make some modifications from these fields to satisfy the users. 展开更多
关键词 Body measurement ANTHROPOMETRY Measuring standard PATTERN
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SPIO-enhanced magnetic resonance imaging for differentiating metastatic from hyperplastic lymph nodes: A study in rabbits
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作者 吴元魁 许乙凯 +3 位作者 黄其鎏 张嘉宁 吴凤林 刘杏元 《Journal of Medical Colleges of PLA(China)》 CAS 2003年第4期218-221,共4页
Objective: To investigate the potential of superparamagnetic iron oxide particles (SPIO) in MR imaging for the differentiation between hyperplastic and metastatic lymph node. Methods: Animal models of malignant lymph ... Objective: To investigate the potential of superparamagnetic iron oxide particles (SPIO) in MR imaging for the differentiation between hyperplastic and metastatic lymph node. Methods: Animal models of malignant lymph node metastasis were established in 6 New-Zealand rabbits by a unilateral intra-muscular injection of VX2 carcinoma cells, and models of hyperplastic lymph nodes were induced in another 6 rabbits by a unilateral intra-muscular injection of egg yolk emulsion. MR images of the lymph nodes were obtained before and 12 h after interstitial injection of SPIO. Image results were analyzed and compared with pathological findings. Results: On unenhanced images, the signal intensity of hyperplastic and metastatic lymph nodes did not differ significantly. After administration of SPIO, the signal intensity of both hyperplastic and metastatic lymph nodes remained unchanged on T1-weighted SE images. On T2-weighted SE images, the signal intensity of hyperplastic lymph nodes decreased heterogeneously, while that of all metastatic ones remained unchanged. On T2-weighted GRE images, the signal intensity of hyperplastic lymph nodes decreased significantly and homogeneously, while that of 4 metastatic ones remained unchanged and that of the rest 2 decreased heterogeneously. Conclusion: SPIO-enhanced MR imaging may enable the differentiation between the hyperplastic and metastatic lymph nodes. 展开更多
关键词 animal laboratory neoplasm metastasis contrast agent lymph node magnetic resonance imaging
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The Study of hemodynamics and coronary arteriography of Patients with Different Types of Unstable Angina Pectoris
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作者 谢小鲁 王燕妮 +1 位作者 祝家庆 袁祖贻 《South China Journal of Cardiology》 CAS 2003年第2期81-86,共6页
Objectives To explore the basic heart functional state and cardiac reserve function of patients with different types of unstable angina pectoris (UAP) and observe the relations between the heart function and severity ... Objectives To explore the basic heart functional state and cardiac reserve function of patients with different types of unstable angina pectoris (UAP) and observe the relations between the heart function and severity of coronary arterial disease. Methods 70 cases with UAP were enrolled including 25 patients with angina decubitus (AD), 23 patients with mixed angina (MA) , and 22 patients with accelerated effort angina (AEA). All patients underwent a series of examination such as UCG, ECT, hemodynamics and volume-loading test. The patients were divided into three groups in light of the results of the hemodynamic examination: ① diastolic dysfunction group ② systolic dysfunction group ③ normal heart function group. We assessed the basic heart function and cardiac reserve function of patients with different types of UAP and also observed the relations between coronary arteriography and heart function. Results ① Under basic conditions, patients with angina decubitus suffered from the systolic (36%) or diastolic dysfunction (48%). 43 percent of the patients with mixed angina had systolic dysfunction and other 43 percent of them had normal cardiac function. However, patients with accelerated effort angina (AEA) were characterized by diastolic dysfunction (31%) or normal heart function (60%). ② In hemodynamic examination, the pulmonary capillary wedge pressure of positive patients rose, at the same time cardiac index fell to the extreme instantaneously after loading in volume-loading test and then they restored to the basic level until 60 minutes. However, both PCWP and CI of the negative patients reach the peak momentarily after loading. They returned to the basic level within 30 minutes. ③ coronary arteriography show: there are 41% of patient with three-vessel disease, 50% with two-vessel disease, 9% single vessel disease and left main narrowing 22. 7% in AEA. There are separately 76%, 24%, 0% and 36% in AD; and 26.1% , 43.4% , 21.7% , 43.4% in MA. ④ Constituent ratio of the AEA and MA were separately compared with AD, AEA/AD: P < 0.05(P = 0.031); MA/AD: P < 0.01 (P = 0.000313). Conclusions Most of patients with the above three types of unstable angina pectoris suffered from the basic heart dysfunction and cardiac reserve dysfunction which might participate in the occurrence and development of unstable angina pectoris. In angiography, there are the most three-vessel diseases in AD that are, therefore, the most severe UAP. 展开更多
关键词 Unstable angina pectoris Hemodynamics Cardiac function Coronary arteriography
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表达绿色荧光蛋白的A型人源呼吸道合胞病毒反向遗传系统的构建
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作者 倪思睿 简书令 +3 位作者 余飞 易若男 武小琰 鲍恩东 《南京农业大学学报》 北大核心 2025年第6期1335-1342,共8页
[目的]为了更好地研究人呼吸道合胞病毒(HRSV)的发病机制并为其应用性研究提供便利,本研究构建了人源HRSV-A2毒株的反向遗传操作系统,并将绿色荧光蛋白(GFP)插入HRSV-A2基因组中,构建表达GFP的重组HRSV-A2。[方法]以细菌人工染色体(BAC... [目的]为了更好地研究人呼吸道合胞病毒(HRSV)的发病机制并为其应用性研究提供便利,本研究构建了人源HRSV-A2毒株的反向遗传操作系统,并将绿色荧光蛋白(GFP)插入HRSV-A2基因组中,构建表达GFP的重组HRSV-A2。[方法]以细菌人工染色体(BAC)为载体骨架构建HRSV-A2毒株全长质粒,并以此为基础在P与M基因之间插入GFP基因;以pCI为载体构建分别表达HRSV N、P、M2-1和L蛋白的4个辅助质粒,并将5个质粒共转染于表达T7 RNA聚合酶的BHK21-T7细胞系进行病毒拯救。[结果]提取rBAC-RSV-A2重组病毒和表达绿色荧光的rBAC-RSV-A2-P-GFP-M重组报告病毒RNA,通过RT-PCR进行全基因组测序,结果表明重组病毒构建成功;在F_(1)—F_(9)连续传代过程中均有绿色荧光表达,F_(10)代荧光消失且无细胞病变;测定重组病毒的生长曲线结果表明,2株重组病毒的生长趋势与原始毒的生长趋势大致相同,表明插入外源基因并不会改变病毒本身的生长特性;噬斑观察及染色结果显示,重组病毒的噬斑小于亲本病毒,表明重组病毒增殖水平下降。[结论]本研究成功构建了以BAC载体为骨架的HRSV-A2感染性克隆,获得了可以表达GFP的重组HRSV病毒。该研究所建立的高效、稳定的HRSV反向遗传学系统,为HRSV致病机制研究、疫苗和抗体药物研发提供了有益的技术平台。 展开更多
关键词 人呼吸道合胞病毒 绿色荧光蛋白 反向遗传系统 重组病毒
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RESPONSES OF THE YOUNG SPIKE OF WHEAT (TRITICUM AESTIVUM) AT DIFFERENT DEVELOPMENTAL STAGES TO CULTURE IN VITRO
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作者 舒理慧 《Chinese Science Bulletin》 SCIE EI CAS 1983年第3期397-400,共4页
Is there any difference between the abilities of differentiation and redifferentiation of tissues of wheat spike in the process of culture in vitro when it is at different developmental stages So far no reports about ... Is there any difference between the abilities of differentiation and redifferentiation of tissues of wheat spike in the process of culture in vitro when it is at different developmental stages So far no reports about this have been found in the literature. 展开更多
关键词 WHEAT DEVELOPMENTAL spike young roots EMPTY inoculated ORGANS FLORAL generative
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XPS STUDY ON DIFFERENT CRYSTALLINE FORMS OF COPPER PHTHALOCYANINE
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作者 王殿勋 石祖荣 《Chinese Science Bulletin》 SCIE EI CAS 1985年第5期612-615,共4页
The copper phthalocyanine (CuPc) as a dye and photoconductive material plays an important role in bionics. The different crystal forms of CuPc have distinct properties. In this article XPS results of α, β, γ, X-CuP... The copper phthalocyanine (CuPc) as a dye and photoconductive material plays an important role in bionics. The different crystal forms of CuPc have distinct properties. In this article XPS results of α, β, γ, X-CuPc have been reported. The character of shake-up satellite peaks and valence band spectra in these CuPc compounds was also examined. It was found that the difference of X-CuPc with other crystal forms of CuPc is obvious. XPS study on different crystal forms of CuPc has not been reported previously. 展开更多
关键词 SHAKE PHTHALOCYANINE VALENCE PHOTOELECTRON unsaturated listed CARLSON deter SUDDEN 加功
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DIFFERENCE SPECTRAL CHARACTERIZATION OF MICROSOMAL CYTOCHROME P-450 FROM LARVAE OF MOSQUITOES(Culex pipiens pallens Coq.)
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作者 沈建华 姜家良 《Chinese Science Bulletin》 SCIE EI CAS 1987年第6期408-412,共5页
Cytochrome P-450 is widely distributed in different species of animals. As a terminal oxidase ot the microsomal monooxygenase system, cytochrome P-450 is the key element of the enzyme system. Because of its broad subs... Cytochrome P-450 is widely distributed in different species of animals. As a terminal oxidase ot the microsomal monooxygenase system, cytochrome P-450 is the key element of the enzyme system. Because of its broad substrate specificity, it is involved in the biotransformation of many xenobiotic and endogenous compounds which are important in envitonmental protection, pharmacology and carcinogenicity. Microsomal P-450 is also a 展开更多
关键词 CULEX pharmacology endogenous CYTOCHROME specificity TROUGH tobacco LARVAE saturation insect
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DIFFERENTIATION OF ENTEROBACTERIA BY ISOTHERMAL PYROLYSIS GAS-LIQUID CHROMATOGRAPHY
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作者 金天如 谢少文 周方 《Chinese Science Bulletin》 SCIE EI CAS 1985年第3期417-420,共4页
A prominent feature for modern biological research is to make full use of different apparatus and techniques, such as spectrometry, chromatography, mass spectrometry and so on, which are hitherto chiefly used in the f... A prominent feature for modern biological research is to make full use of different apparatus and techniques, such as spectrometry, chromatography, mass spectrometry and so on, which are hitherto chiefly used in the fields of chemistry and physics. Gasliquid chromatography, as a new method for analytical chemistry, was invented by 展开更多
关键词 apparatus distilled hundred entero MINUTE PROTEUS indie eared stored NUTRIENT
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DIFFERENCE SCHEMES FOR FULLY NONLINEAR PSEUDO-HYPERBOLIC SYSTEMS 被引量:1
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作者 周毓麟 杜明笙 《Acta Mathematicae Applicatae Sinica》 SCIE CSCD 1991年第4期298-320,共23页
The general difference schemes for the first boundary problem of the fully nonlinear pseudo-hyperbolic systemsf(x, t, u,u_x,u_(xx),u_t,u_(tt),u_(xt),u_(xxt))=0are considered in the rectangular domain Q_T={0≤x≤1, 0≤... The general difference schemes for the first boundary problem of the fully nonlinear pseudo-hyperbolic systemsf(x, t, u,u_x,u_(xx),u_t,u_(tt),u_(xt),u_(xxt))=0are considered in the rectangular domain Q_T={0≤x≤1, 0≤t≤T}, where u(x,t)and f(x, t, u,p_1, p_2, r_1,r_2,q_1,q_2) are two m-dimensional vector functions with m≥1 for(x, t)∈Q_T and u,p_1,p_2,r_1,r_2,q_1,q_2∈R. The existence and the estimates of solutionsfor the finite difference system are established by the fixed point technique. The absolute andrelative stability and convergence of difference schemes are justified by means of a series of a prioriestimates. In the present study, the existence of unique smooth solution of the original problemis assumed. The similar results for nonlinear and quasilinear pseudo-hyperbolic systems are alsoobtained. 展开更多
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SAMPLING OF THE VISUAL INFORMATION BY THE NEURAL SUPERPOSITION COMPOUND EYES IN DIFFERENT ADAPTED STATES
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作者 吴卫国 郭爱克 +3 位作者 冯春华 石淑珍 王谷岩 吴盛立 《Chinese Science Bulletin》 SCIE EI CAS 1985年第5期668-671,共4页
Ⅰ. JNTRODUCTION Each ommatidium in the diptera compound eye contains 8 retinular cells. The first subsystem consists of 6 large retinular cells in periphery. Six different peripheral retinular cells from the 6 neighb... Ⅰ. JNTRODUCTION Each ommatidium in the diptera compound eye contains 8 retinular cells. The first subsystem consists of 6 large retinular cells in periphery. Six different peripheral retinular cells from the 6 neighbouring ommatidia receive visual information and transmit it into a single neuro-ommatidium (cartridge) of the first optic ganglion. The other two retinular cells are located at the centre of the ommatidium from the second subsystem, which receives visual information and sends it directy to the second optic ganglion. 展开更多
关键词 VISUAL transmit subsystem optic GANGLION PERIPHERY centre DISTAL PIGMENT elsewhere
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Laser-induced damage of 1064-nm narrow-band interference filters under different laser modes
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作者 高卫东 贺洪波 +1 位作者 邵建达 范正修 《Chinese Optics Letters》 SCIE EI CAS CSCD 2004年第8期493-494,共2页
The laser-induced damage behavior of narrow-band interference filters was investigated with a Nd:YAG laser at 1064 nm under single-pulse mode and free-running laser mode. The absorption measurement of such coatings ha... The laser-induced damage behavior of narrow-band interference filters was investigated with a Nd:YAG laser at 1064 nm under single-pulse mode and free-running laser mode. The absorption measurement of such coatings has been performed by surface thermal lensing (STL) technique. The relationship between damage morphology and absorption under the two different laser modes was studied in detail. The explanation was given by the standing-wave distribution theory. 展开更多
关键词 Laser modes Light interference Neodymium lasers Optical filters
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Simultaneously compensating tunable CD and adaptive PMD using two different nonlinearly chirped fiber Bragg gratings
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作者 于丽 张晓光 +4 位作者 周光涛 刘玉敏 张建忠 席丽霞 杨伯君 《Chinese Optics Letters》 SCIE EI CAS CSCD 2004年第8期445-448,共4页
Experimental research of simultaneous tunable chromatic dispersion (CD) and adaptive polarization mode dispersion (PMD) compensations in optical fiber communication system was reported. Two different nonlinearly chirp... Experimental research of simultaneous tunable chromatic dispersion (CD) and adaptive polarization mode dispersion (PMD) compensations in optical fiber communication system was reported. Two different nonlinearly chirped fiber Bragg gratings fabricated through the equivalent chirp technology were adopted in the experiment. One of the gratings was used as CD compensator, with a tunable dispersion range from 300 to 600 ps/nm. The other made of photosensitive polarization maintaining fiber was used as a tunable delay line of PMD compensator, which provided a varying amount of differential group delay (DGD) from 40 to 110 ps. Our experiment was operated at 10-Gb/s non-return-to-zero (NRZ) system and the results showed that the eye pattern recovery is excellent after both PMD and CD are compensated. Especially, the power penalty at a bit error rate (BER) of 10-9 is about 1 dB. 展开更多
关键词 Bit error rate Electromagnetic dispersion Fiber Bragg gratings
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猫杯状病毒反向遗传系统的建立及弱毒株的构建 被引量:2
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作者 于雯 孙慧敏 +2 位作者 方晨捷 宋家升 鲍恩东 《畜牧与兽医》 CAS 北大核心 2024年第5期107-112,共6页
为了构建猫杯状病毒(FCV)弱毒疫苗候选株,从杭州流行的FCV毒株FCV-HZ-DF-19中提取基因组RNA,利用RT-PCR技术分3段扩增获得FCV全基因组,通过同源重组的方法将全长基因组克隆至已插入榔头状核酶(HamRz)和丁型肝炎核酶(HdvRz)的PCI载体,获... 为了构建猫杯状病毒(FCV)弱毒疫苗候选株,从杭州流行的FCV毒株FCV-HZ-DF-19中提取基因组RNA,利用RT-PCR技术分3段扩增获得FCV全基因组,通过同源重组的方法将全长基因组克隆至已插入榔头状核酶(HamRz)和丁型肝炎核酶(HdvRz)的PCI载体,获得中间质粒pPCI-FCV;以pPCI-FCV为模板扩增获得含HamRz、HdvRz和FCV全基因组的产物FCV-H,并将其连接到BAC载体中获得含有FCV全基因组的克隆pBAC-FCV。将pBAC-FCV转染猫肾细胞F81连续传代后收获病毒。针对FCV毒力相关的LC基因进行缺失,获得感染性克隆pBAC-FCV-ΔLC,通过转染表达FCV-VP1蛋白的猫肾细胞系进行病毒拯救。结果显示,拯救毒株rBAC-FCV和LC基因缺失毒株rBAC-FCV-ΔLC均可产生细胞病变;测序结果表明,拯救毒株的基因序列与亲本毒株FCV-HZ-DF-19一致,且生长曲线与亲本毒相似,而基因缺失毒株的生长则慢于亲本毒株,产生的噬斑大小也明显小于亲本毒株,表明缺失LC基因后病毒毒力下降。结论:本研究成功构建了FCV的反向遗传系统及基因缺失弱毒株,为后续弱毒疫苗研发和FCV生物学特性研究奠定了基础。 展开更多
关键词 猫杯状病毒 反向遗传学技术 同源重组 基因缺失
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猫疱疹病毒Ⅰ型gIgE基因缺失毒株的构建及生物学特性分析 被引量:1
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作者 张晶晶 孙慧敏 +1 位作者 宋家升 鲍恩东 《南京农业大学学报》 CAS CSCD 北大核心 2024年第3期515-521,共7页
[目的]为了开发一种能够预防感染、阻止潜伏期建立的猫疱疹病毒Ⅰ型(FHV-1)弱毒疫苗,利用同源重组以及CRISPR/Cas9技术将FHV-1的gIgE基因替换为红色荧光蛋白基因(RFP)。[方法]构建针对gIgE基因的3条sgRNA及表达Cas9蛋白的质粒,将含有sg... [目的]为了开发一种能够预防感染、阻止潜伏期建立的猫疱疹病毒Ⅰ型(FHV-1)弱毒疫苗,利用同源重组以及CRISPR/Cas9技术将FHV-1的gIgE基因替换为红色荧光蛋白基因(RFP)。[方法]构建针对gIgE基因的3条sgRNA及表达Cas9蛋白的质粒,将含有sgRNA及Cas9蛋白的质粒与含有RFP的转移载体共转染到猫肾细胞(F81),通过噬斑纯化以及有限稀释方法进行病毒纯化,以重组病毒感染F81细胞连续10代验证其遗传稳定性,通过测定重组病毒以及亲本病毒H07的病毒滴度、一步生长曲线以及判断噬斑大小评价其生物学特性。[结果]成功构建了FHV-1 gIgE基因缺失毒株并将其命名为FHV-ΔgIgE-RFP;提取重组病毒DNA,通过PCR验证gIgE基因无目的条带,表明重组病毒纯化成功,再通过PCR验证RFP基因有目的条带,表明重组病毒成功插入外源基因。病毒滴度测定结果显示,与亲本病毒相比重组病毒的病毒滴度下降了90%;在F 1—F 10连续传代过程中RFP荧光表达且无减弱现象,表明重组病毒可稳定表达外源基因;测定FHV-ΔgIgE-RFP以及H07的生长曲线,重组病毒的生长趋势与亲本病毒大致相同,表明插入外源基因并不会改变病毒本身的生长特性;噬斑观察及染色结果显示,在病毒感染细胞前、后期,重组病毒的噬斑都小于亲本病毒,表明缺失gIgE基因后病毒毒力下降。[结论]成功构建并筛选出FHV-1 gIgE基因缺失致弱毒株,为开发疱疹病毒弱毒活载体疫苗奠定了基础。 展开更多
关键词 猫疱疹病毒Ⅰ型(FHV-1) 致弱毒株 重组病毒 病毒筛选
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