In vitro effect of p2l^(WAF-1/CIP1) gene on growth of human glioma cells mediated by EGFR targeted non-viral vector GE7 system

To construct the EGFR targeted non-viral vector GE7 system and explore the in vitro effect of p21^WAF-1/CIP1 gene on growth of human glioma cells mediated by the GE7 system. Methods: The EGFR targeted non-vi-ral vector GET gene delivery system was constructed. The malignant human glioma cell llne U251MG was transfected in vitro with β-galactosidase gene( reporter gene) and p21^WAF-1/CIP1 gene (therapeutic gene) using the GET system. By means of X-gal staining, MTS and FACS, the transfection efficiency of exogenous gene and apoptosis rate of tumor cells were examined. The expression of p21^WAF-1/CIP1 gene in transfected U251MG cell was examined by immunohistochemitry staining. Results: The highest transfer rate of exogenous gene was 70%. After transfection with p21^WAF-l/CIP1 gene,the expression of WAF-1 increased remarkably and steadily; the growth of U251MG cells were inhibited evidently.FACS examination showed G1 arrest. The average apoptosis rate was 25.2%. Conclusion: GE7 system has the ability to transfer exogenous gene to targeted cells efficiently, and expression of p21^WAF-l/CIP1 gene can induce apoptosis of glio-ma cell and inhibit its growth.

Non-viral targeted delivery system mediates transfection of thymidine kinase gene of herpes simplex virus into ovarian cancer cells:a comparison between one time and continuous mediation

Objective: To compare the transferring efficiency and killing effects of one time and continuous mediation with GE7,a non-viral targeted delivery system,in transfection of thymidine kinase gene of herpes simplex virus (HSV-tk) into ovarian cancer cells. Methods: GE7 was used to prepare recombinants with β-galactosidase (β-gal) and HSV1-tk; the re-combinants were then used to transfect human ovarian cancer line CaOV3 once and continuously. β-gal staining was used to compare the efficiencies of one time and continuous mediation with GE7 system. Ganciclovior (GCV) was introduced into HSV1-tk transfected ovarian cells. Through drawing the cell growth curve and flow cytometry,the killing effects of GCV on once and continuously GE7/HSV1-tk transfected cells were observed. Results: We found that the one time and continuous exogenous gene transfer efficiencies were about 80% and 85%,respectively. When 1μg/mL GCV was used to treat ovarian cell transfected with HSV1-tk gene,growth inhibiting rates of ovarian cells of one time and continuous transferring were 82% and 90%,respectively; their apoptosis indices were 15 and 30,respectively. Under same GCV concentration,continuous me-diation of GE7/pCMV-tk transfection into ovarian cancer cells had more significant inhibitory effect than one time mediation (P<0.05). Conclusion: Compared with one time mediation,continuous mediation of transfection with GE7 gene delivery system has higher efficiency. Continuous mediation of GE7/HSV1-tk/GCV therapeutic gene system has more powerful killing effect.

A RGD-Containing Oligopeptide (K)_(16)GRGBSPC: A Novel Vector for Integrin-Mediated Targeted Gene Belivery

A 23 amino acid, bifunctional integrin-targeted synthetic oligopeptide was evaluated for ex vivo gene delivery to rabbit bone marrow stromal cells （BMSCs）. Synthesis of the peptide （K）16GRGDSPC was performed on a solid-phase batch peptide synthesizer. BMSCs were transfected with plasmid DNA coding for luciferase by （K）j6GRGDSPC and the transfection efficiency was assayed. The influences of chloroquine and polyethyleneimine on the transfection efficiency were also examined. The target specificity of （K）16GRGDSPC to mediate exogenous gene into BMSCs was analyzed using cell attachment test and gene delivery inhibition test. The results showed that the transfection efficiency of the oligopeptide vector was lower than that of Lipofectamine. But in the presence of endosomal buffer chloroquine or endosomal disrupting agent polyethyleneimine, the transfection efficiency of the vector was greatly enhanced. In addition, RGD-containing peptides inhibited BMSCs＇ attachment to the 96-well plates pretreated with fibronectin or vitronecfin and significantly decreased the transfection efficiency of the oligopeptide vector. These studies demonstrated that oligopeptide （K）16GRGDSPC was an ideal novel targeted non-viral gene delivery vector, which was easy to be synthesized, high efficient and low cytotoxicity. The vector could effectively deliver exogenous gene into rat BMSCs.

Surface Modification of Biomimetic PLGA-(ASP-PEG) Matrix with RGD-Containing Peptide:a New Non-Viral Vector for Gene Transfer and Tissue Engineering

RGD-containing peptide （ K16-GRGDSPC） , characterized as non-viral gene vectors, was fabricated to modify the surface of PLGA-[ASP- PEG] matrix, which offered the foundation for gene transfer with porous matrix of gene activated later. Peptide was synthesized and matrix was executed into chips A, B and chip C. Chip C was regarded as control. Chips A and B were reacted with cross-linker. Then chip A was reacted with peptide. MS and HPLC were ased to detect the .14W and purity of peptide. Sulphur, existing on the surface of biomaterials, was detected by XPS. The purity of un-reacted peptide in residual solution was detected by a spectrophotometer. HPLC shows that the peptide purity was 94%- 95% , and MS shows that the MW was 2 741. 3307. XPS reveals that the binding energy of sulphur was 164 eV and the ratio of carbon to sulphur （C/S） was 99. 746 ：0. 1014 in reacted chip A. The binding energy of sulphur in reacted chip B was 164 eV and 162 eV, C/ S was 99.574：0.4255, aM there was no sulphur in chip C. Peptide was manufactured and linked to the surface of biomimetic and 3-D matrix, which offered the possibilities for gene transfer and tissue engineering with this new kind of non-viral gene vector.

Targeting lentiviral vectors to primordial germ cells(PGCs):An efficient strategy for generating transgenic chickens

Recent advances in avian transgenic studies highlight the possibility of utilizing lentiviral vectors as tools to generate transgenic chickens. However, low rates of gonadal chimerism and germ line transmission efficiency still limit the broad usage of this method in creating transgenic chickens. In this study, we implemented a simple strategy using modified lentiviral vectors targeted to chicken primordial germ cells(PGCs) to generate transgenic chickens. The lentiviral vectors were pseudotyped with a modified Sindbis virus envelope protein(termed M168) and conjugated with an antibody specific to PGC membrane proteins. We demonstrated that these optimized M168-pseudotyped lentiviral vectors conjugated with SSEA4 antibodies successfully targeted transduction of PGCs in vitro and in vivo. Compared with the control, 50.0%–66.7% of chicken embryos expressed green fluorescent protein(GFP) in gonads transduced by the M168-pseudotyped lentivirus. This improved the targeted transduction efficiency by 30.0%–46.7%. Efficient chimerism of exogenous genes was also observed. This targeting technology could improve the efficiency of germ line transmission and provide greater opportunities for transgenic poultry studies.

Non-viral liposome-mediated transfer of brain-derived neurotrophic factor across the blood-brain barrier

Brain-derived neurotrophic factor（BDNF） plays an important role in the repair of central nervous system injury,but cannot directly traverse the blood-brain barrier.Liposomes are a new type of non-viral vector,able to carry macromolecules across the blood-brain barrier and into the brain.Here,we investigate whether BDNF could be transported across the blood-brain barrier by tail-vein injection of liposomes conjugated to transferrin（Tf） and polyethylene glycol（PEG）,and carrying BDNF modified with cytomegalovirus promoter（pC MV） or glial fibrillary acidic protein promoter（p GFAP）（Tf-p CMV-BDNF-PEG and Tf-p GFAP-BDNF-PEG,respectively）.Both liposomes were able to traverse the blood-brain barrier,and BDNF was mainly expressed in the cerebral cortex.BDNF expression in the cerebral cortex was higher in the Tf-p GFAP-BDNF-PEG group than in the Tf-p CMV-BDNF-PEG group.This study demonstrates the successful construction of a non-virus targeted liposome,Tf-p GFAP-BDNF-PEG,which crosses the blood-brain barrier and is distributed in the cerebral cortex.Our work provides an experimental basis for BDNF-related targeted drug delivery in the brain.

Automatic target recognition of moving target based on empirical mode decomposition and genetic algorithm support vector machine

In order to improve measurement accuracy of moving target signals, an automatic target recognition model of moving target signals was established based on empirical mode decomposition(EMD) and support vector machine(SVM). Automatic target recognition process on the nonlinear and non-stationary of Doppler signals of military target by using automatic target recognition model can be expressed as follows. Firstly, the nonlinearity and non-stationary of Doppler signals were decomposed into a set of intrinsic mode functions(IMFs) using EMD. After the Hilbert transform of IMF, the energy ratio of each IMF to the total IMFs can be extracted as the features of military target. Then, the SVM was trained through using the energy ratio to classify the military targets, and genetic algorithm(GA) was used to optimize SVM parameters in the solution space. The experimental results show that this algorithm can achieve the recognition accuracies of 86.15%, 87.93%, and 82.28% for tank, vehicle and soldier, respectively.

Support vector regression model for complex target RCS predicting

The electromagnetic scattering computation has developed rapidly for many years; some computing problems for complex and coated targets cannot be solved by using the existing theory and computing models. A computing model based on data is established for making up the insufficiency of theoretic models. Based on the ＂support vector regression method＂, which is formulated on the principle of minimizing a structural risk, a data model to predicate the unknown radar cross section of some appointed targets is given. Comparison between the actual data and the results of this predicting model based on support vector regression method proved that the support vector regression method is workable and with a comparative precision.

Hepatitis B virus envelope as a targeting gene transfer vector for hepatic cancer cells

学习的目的是观察肝炎 B 病毒信封(HBVE ) 的 transfection 功效并且为肝癌症房间作为基因转移向量评估它的能力的目的。获得 HBVE 的方法， HepG 2.2.15 房间的上层清液液体与集中的一个 PEG8000 解决方案被混合并且被 β-propiolactone 使失去活性。获得的 HBVE 被用来包装 pIRES 2-EGFP 测试它的包裹能力。然后，我们与 ELISA， PCR， SDS 页和电子显微镜学检验了它的数量和质量。pIRES 2-EGFP 与 HBVE 被包装并且获得了产品 HBVE-GFP。pIRES 2-EGFP 被包装与脂肪一些并且获得了产品 liposome-GFP。HBVE-GFP 和 liposome-GFP 被用来转移 HepG 2 房间学习 transfection 效率。HBVE-GFP 被用来转移学习指向的能力的 HepG 2， A549， HeLa 和 FB 房间。绿荧光灯的蛋白质(GFP ) 表示在一台荧光灯的显微镜下面被观察。GFP 积极房间的率被流动 cytometry 决定。结果 1。获得的 HBVE 能保留表面蛋白质 HBsAg + pre S1 + pre S2 并且没有病毒 DNA。它为 pIRES 2-EGFP 有好包裹能力。2。Transfection 效率：GFP 能在荧光灯的显微镜下面在脂肪的一些组和 HBVE 组被观察。但是 HBVE 组比脂肪的一些组有更高荧光灯的紧张。脂肪的一些组的 transfection 率是 49.97% ± 2.37% 当 HBVE 组是 70.65% ±时 3.15% 并且 HBVE 组的荧光灯的紧张是 3 4 次(P = 0.000 ) 为有流动 cytometry 的决心的脂肪的一些组。3。指向能力：GFP 能在荧光灯的显微镜下面在四个组被观察。HepG 2 组在四个组之中有最高荧光灯的紧张。HepG 2 组的 transfection 率是 71.35% ± 0.03% 它高度比另外的组被表示(P = 0.000 ) 并且 HepG 2 组的荧光灯的紧张是 2 3 次(P = 0.000 ) 为有流动 cytometry 的决心的另外的 3 个组。结论 HBVE 能从 HepG 2.2.15 房间的上层清液液体与 PEG8000 和 β-propiolactone 的方法成功地被构造。HBVE 能是为肝癌症房间的候选人基因转移向量。

Construction and application of gene targeting replacement vector formouse coagulation factor IX gene

Objective: To construct and apply a replacement targeting vector for mouse coagulation factor IX(mFIX) gene in embryonic stem (ES) cells. Methods: Based on the cloning and structural analysis of the genomic DNA fragment of coagulation factor IX gene from 129Sv mouse genomic DNA A phage library, PMFIXDEL plasmid was designed with positive-negative-selection (PNS) strategy, and constructed with commonmolecular cloning techniques. Structure of PMFIXDEL was identified by PCR and restriction analysis. Afterelectroporation with the linearized PMFIXDEL DNA, transfected ES cells were cultured in G418/GANC drugselection medium. The recombination efficacy of this vector was tested. Results: The main components ofPMFIXDEL were two copies of negative selection gene (HSV-tk expression cassette), a positive selectiongene (Neo expression cassette), long and short homologous fragments and plasmid backbone. The introduction of negative selection gene (HSV-tk ) into the construct resulted in 24-fold increase of selection.Conclusion: An effective replacement vector for mFIX gene targeting was constructed and applied in ES cell.

基于SVM算法的虚假航迹识别

针对云雨杂波和主被动干扰导致多雷达传感器产生虚假目标航迹的问题,利用支持向量机(SVM)算法的自主学习能力,通过构建基于数据驱动的判别模型进行虚假航迹识别。针对航迹起始得到的目标潜在航迹,利用人工智能数据驱动、自学习的特点,设计了SVM算法。通过对已标记真假的目标航迹样本进行离线学习,形成虚假航迹识别的SVM分类器,实现了基于数据驱动的判别模型代替先验知识规则约束的固定模型,并在工程应用中,利用SVM分类器在线识别虚假航迹,完成实时剔除。通过实测雷达数据实验验证,该算法的目标虚假航迹准确率高达95%以上,完全满足实际的工程应用需求。相比基于阈值或规则进行硬性判断的传统虚假航迹识别方法,所提出的算法不仅提高了准确率,还具有较高的实时性,能够适应复杂多变的杂波环境,在实际应用中具有更强的适应性和实用性。因此,提出的基于SVM算法的虚假航迹识别方法对于密集杂波场景下的虚假航迹剔除问题具有显著的实际应用价值。

基于状态可观测性和多模态数据PF的移动目标跟踪

为了实现对移动目标的跟踪,提出了一种新颖的基于目标状态可观测性和多模态数据粒子滤波(Particle Filter,PF)的跟踪方案。通过部署在目标移动区域中的传感器获得跟踪目标的距离和到达方向测量值,对接收到的数据进行预处理来计算PF的观测值,以形成一个临时距离图像。通过利用状态更新函数和形成的候选图像模板确定目标状态向量;在PF器中加入额外的加权阶段,使得PF器可自适应地同步多模态数据流,以实现鲁棒的目标跟踪。仿真实验结果验证了所提方案能够有效地跟踪移动目标。

基于卷积神经网络的红外弱小车辆目标检测方法

传统方法无法获得理想的红外弱小车辆目标检测结果,导致检测误差大,无法满足实际应用要求,为了解决传统红外弱小车辆目标检测方法存在的局限性,及时检测红外图像中的弱小车辆,提高车辆检测精度,设计了基于卷积神经网络的红外弱小车辆目标检测方法。首先对弱小车辆目标检测需要的红外图像进行采集,并对红外图像噪声进行处理,消除噪声对弱小车辆目标检测的干扰,然后采用卷积神经网络建立弱小车辆目标检测模型,最后通过具体仿真实验测试弱小车辆目标检测方法的性能。结果表明,该方法的弱小车辆目标检测精度超过了90%,大幅度减少了弱小车辆目标的误检率,同时弱小车辆目标检测时间控制在5 s内,可以满足弱小车辆目标检测的实时性要求,具有较高的实际应用价值。

基于正则化正交非负矩阵分解的旋转目标检测方法

小样本的旋转目标检测是指在样本数少的情况下进行旋转目标检测模型的训练,深度学习在旋转目标检测领域往往需要庞大的样本数和计算算力。现有的基于机器学习的旋转目标检测方法大多有着对目标尺度和姿态敏感的缺点。因此提出一种基于正则化正交非负矩阵分解的旋转目标检测方法,来解决小样本的旋转目标检测难题。首先,针对样本不具有各种角度的图片,对样本进行旋转后进行背景填充,这样便于更好的表征学习。其次,提出一种基于正则化正交非负矩阵分解算法对旋转样本的梯度直方图特征进行表征学习。最后,为了测试算法在特征学习后的有效性,利用支持向量机对特征提取后的数据进行训练和测试。实验结果表明本文的目标检测方法在多个数据集中可以取得不错的效果。

基于遗传算法优化支持向量机的船舰目标识别分类

为了实现有效的海上监管和响应,提高舰船监管效率,降低人力成本,提出基于遗传算法优化支持向量机的舰船目标识别分类方法。以HU矩为舰船目标的特征描述子,在舰船目标图像内,提取具备旋转、尺度与平移不变性的舰船目标特征矩;利用遗传算法,优化支持向量机的惩罚因子与核参数;在参数优化后的支持向量机内,输入舰船目标特征矩样本,输出舰船目标识别分类结果。实验证明,该方法可有效提取舰船目标特征矩;经过参数优化后的支持向量机,可有效降低计算复杂度,加快检测目标识别分类效率,具备较优的舰船目标识别分类性能。该方法均可精准识别分类舰船目标。

一种基于航迹特征的无人机与飞鸟目标雷达识别方法

对于现代雷达探测系统而言,无人机与飞鸟同属于具有“低慢小”特征的一类典型目标,而面对比较复杂的作战环境,其对功能的要求已经不仅局限于对两者目标实现稳定探测跟踪,如何有效区分两者类型并完成识别更是当下急迫且重要的难题。常规方法是从目标的微动特征差异进行区分,但由于两者回波微弱,很难通过时频分析方法提取目标特征。针对该问题,从航迹特征出发,提出一种无人机与飞鸟目标雷达识别方法。首先对比两者目标在运动轨迹上的差异性,进行特征分析,提出时间相关的航向震荡频率与速度震荡频率特征量描述方法,并在离线状态下,利用实测雷达系统记录的航迹数据,提取两者的有效特征量;然后利用支持向量机算法对样本进行训练,并在获得最优模型参数后,通过测试样本进行测试,测试分类结果显示准确识别率能够达87%;最后在线状态下跟飞实验,其结果既表明该方法的正确性,也体现了在工程实现角度上的轻量性、实用性、适用性,具有较高价值。

基于ISVM-DS的红外多传感器融合识别方法

弹道中段目标为一个目标群,包括弹头、诱饵、碎片等,并且由于距离传感器较远,红外成像为点目标,可用信息较少,因此单一的红外传感器往往难以满足识别要求,需要融合多个传感器进行识别。针对红外多传感器的融合识别问题,本文提出了基于增量支持向量机和D-S(increment support vector machine-Dempster-Shafer,ISVM-DS)证据理论的融合识别方法。首先,训练多个波段传感器红外特征的支持向量数据描述(support vector data description,SVDD)模型,生成壳向量并训练其ISVM模型;接着,采用ISVM模型的后验概率生成基本概率赋值(basic probability assignment,BPA);最后,利用D-S证据理论对多个证据的BPA进行融合,输出分类结果。实验结果表明,该方法能有效提高目标识别的准确性。

室内火灾高浓度烟雾环境火点增强识别仿真

火灾火点在形态上均具有不确定性,尤其在高浓度烟雾火灾环境中火点还具有随机透明度,易与图像的背景部分混合,导致火点位置识别难度较高。为此,提出室内火灾高浓度烟雾环境火点增强识别方法。对高浓度烟雾图像去噪、锐化以及分割处理,完成目标区域轮廓的提取。基于此,提取目标轮廓并对其增强,获取目标区域中火点的特征向量,结合孪生支持向量机对火点特征展开分类,实现室内火灾高浓度烟雾环境的火点的精准识别。实验结果表明,上述方法的火点识别精度高于98%,耗时低于210ms,且能够有效提取火点目标特征,证明了研究方法的应用效果更好,可靠性更高。

基于时序向量相似性的空间目标群匹配技术研究

分析了空间低轨目标群的运行特点,提出了基于时序向量相似性的空间目标群匹配算法,提高了对低轨巨型星座的识别管理能力。首先,介绍了时序向量的降维方法,将目标群高维观测时序向量简化为空间构型序列;而后,提出了基于动态时间规整(Dynamic Time Warping,DTW)的目标群空间构型序列相似性判别算法;最后,利用星链卫星目标群仿真和实测数据对算法的匹配能力进行验证。结果表明该算法可实现空间目标群监测数据快速匹配,仿真数据匹配过程中,在群内目标缺失30%的条件下匹配成功率可达100%,在低缺失条件下(缺失率5%以内)群内目标识别成功率平均超过75%;实测数据匹配成功率可达100%。

Vector cleaner:一种新的去除测序目的基因载体序列的方法

Sanger测序法测序目的基因常包含有目的基因和载体序列,为了快速去除测序目的基因载体序列,提出了一种新的目的基因载体序列去除方法并开发了程序Vector cleaner。首先利用该程序批量读取引物信息和目的基因测序序列;其次,程序在所读取的引物序列上建立引物半长的滑动窗口来产生种子,通过计数种子与测序序列的匹配次数,定位引物位置和删除引物两侧的载体序列;最后,程序通过比较上游引物序列和其反向互补序列分别与测序序列匹配种子数,判断和转换正义链。使用Vector cleaner对12条GhVIN1基因测序序列进行去载体测试,并与Seqclean和SeqMan软件相比较。结果表明:Vector cleaner能有效去除棉花GhVIN1基因测序载体序列,识别并翻译反义链序列。与Seqclean和SeqMan软件相比较,Vector cleaner正确率高,敏感性强。Vector cleaner、SeqMan和Seqclean所测试序列的总序列数正确率分别为100%、100%和91.6%,总碱基正确率分别为99.90%、99.00%和94.33%。与同类软件比较,Vector cleaner更适合实验人员批量去除测序目的基因载体序列,具有准确率高、敏感性强、自动翻译反义链的特点。