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An efficient and universal protoplast-based transient gene expression system for genome editing in Brassica crops

Horticultural Plant Journal 2024年第4期10卷 983-994页

作者： Xiaoxiao Yu Jintai Yu Yin Lu Wenjing Li guanzhong Huo Jun zhang Yan Li Jianjun Zhao Jun Li State Key Laboratory of North China Crop Improvement and Regulation College of Life SciencesHebei Agricultural UniversityBaodingHebei 071001China Hebei Key Laboratory of Plant Physiology and Molecular Pathology Hebei Agricultural UniversityBaodingHebei 071001China Key Laboratory of Vegetable Germplasm Innovation and Utilization of Hebei Collaborative Innovation Center of Vegetable Industry in HebeiCollege of HorticultureHebei Agricultural UniversityBaodingHebei 071001China College of Modern Science and Technology Hebei Agricultural UniversityBaodingHebei 071001China Human Resource Department Hebei Agricultural UniversityBaodingHebei 071001China

Protoplast-based transient gene expression system has been widely used in plant genome editing because of its simple operation and less *** order to establish a universal protoplast-based transient transfection system for verifying activities of genome editing vectors containing targets in Brassica,we systematically optimized factors affecting protoplast isolation and transient gene *** established an efficient protoplast-based transient gene expression system(PTGE)in Chinese cabbage,achieving high protoplast yield of 4.9×10^(5)·g^(-1)FW,viability over 95%,and transfection efficiency of 76%.We showed for the first time that pretreatment of protoplasts with a hypotonic MMG could significantly enhance the transfection ***,protoplasts incubated at 37℃ for 6 min improved the transfection efficiency to 86%.We also demonstrated that PTGE worked well(more than 50%transfection efficiency)in multiple Brassica species including cabbage,Pak Choi,Chinese kale,and ***,PTGE was used for validating the activities of CRISPR/Cas9 vectors containing targets in Chinese cabbage,cabbage,and pak choi,demonstrating the broad applicability of the established PTGE for genome editing in Brassica crops.

关键词： Chinese cabbage PTGE Transfection efficiency CRISPR/Cas9

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Expanding the targeting scope of CRISPR/Cas9-mediated genome editing by Cas9 variants in Brassica

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aBIOTECH 2024年第2期5卷 202-208页

作者： Wenjing Li Xuan Li Chunyang Wang guanzhong Huo Xinru zhang Jintai Yu Xiaoxiao Yu Jing Li Chao zhang Jianjun Zhao Yan Li Jun Li State Key Laboratory of North China Crop Improvement and Regulation College of Life SciencesHebei Agricultural UniversityBaoding 071001China Hebei Key Laboratory of Plant Physiology and Molecular Pathology Hebei Agricultural UniversityBaoding 071001China Key Laboratory of Vegetable Germplasm Innovation and Utilization of Hebei Collaborative Innovation Center of Vegetable Industry in HebeiCollege of HorticultureHebei Agricultural UniversityBaoding 071001China College of Modern Science and Technology Hebei Agricultural UniversityBaoding 071001China

CRISPR/Cas9,presently the most widely used genome editing technology,has provided great potential for functional studies and plant ***,the strict requirement for a protospacer adjacent motif(PAM)has hindered the application of the CRISPR/Cas9 system because the number of targetable genomic sites is ***,the engineered variants Cas9-NG,SpG,and SpRY,which recognize non-canonical PAMs,have been successfully tested in plants(mainly in rice,a monocot).In this study,we evaluated the targeted mutagenesis capabilities of these Cas9 variants in two important Brassica vegetables,Chinese cabbage(Brassica rapa ***)and cabbage(Brassica oleracea ***).Both Cas9-NG and SpG induced efficient mutagenesis at NGN PAMs,while SpG outperformed Cas9-NG at NGC and NGT *** achieved efficient editing at almost all PAMs(NRN[NYN),albeit with some self-targeting activity at transfer(T)-DNA *** SpRY-induced mutants were detected in cabbage plants in a PAM-less ***,an adenine base editor was developed using SpRY and TadA8e deaminase that induced A-to-G conversions within target sites using non-canonical ***,the toolboxes developed here induced successful genome editing in Chinese cabbage and *** work further expands the targeting scope of genome editing and paves the way for future basic research and genetic improvement in Brassica.

关键词： Brassica Cas9 variants CRISPR Genome editing Non-canonical PAM

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Adaptive　Multisensor　Tracking　Fusion　Algorithm　for　Air－borne　Distributed　Passive　Sensor　Network

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Journal of Systems Engineering and Electronics 1996年第3期7卷 15-23页

作者： Zhen Ding Hongcai zhang & guanzhong Dai (Department of Automatic Control, Northwestern Polytechnical UniversityShaanxi, Xi’an 710072, P.R.China) Department of Automatic Control Northwestern Polytechnical University Shaanxi Xi'an 710072 P.R. China

Single passive sensor tracking algorithms have four disadvantages: bad stability, longdynamic time, big bias and sensitive to initial conditions. So the corresponding fusion algorithm results in bad performance. A new error analysis method for two passive sensor tracking system is presented and the error equations are deduced in detail. Based on the equations, we carry out theoretical computation and Monte Carlo computer simulation. The results show the correctness of our error computation equations. With the error equations, we present multiple 'two station'fusion algorithm using adaptive pseudo measurement equations. This greatly enhances the tracking performance and makes the algorithm convergent very fast and not sensitive to initial *** results prove the correctness of our new algorithm.

关键词： Passive tracking system Error analysis Fusion algorithm Distributed passive sensornetwork Distributed estimation.

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Efficacy of pemetrexed combined with erlotinib/gefitinib in advanced non-small cell lung cancer patients during tyrosine kinase inhibitor treatment

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Oncology and Translational Medicine 2017年第3期3卷 93-98页

作者： guanzhong zhang Zhaozhe Liu Tao Han Xiaodong Xie Shunchang Jiao Department of Oncology General Hospital of Shenyang Military Region Department of Oncology Chinese PLA General Hospital

Objective We aimed to evaluate the efficacy and safety of pemetrexed combined with erlotinib/gefitinib in advanced non-small cell lung cancer(NSCLC) patients during tyrosine kinase inhibitor(TKI) treatment. Methods Thirty-two patients with advanced NSCLC were divided into two groups. Patients in the control group received continuous daily epidermal growth factor receptor tyrosine kinase inhibitor(EGFRTKI) treatment, and patients in the experimental group received continuous daily EGFR-TKI along with pemetrexed treatment, which was administered on day 1 at 500 mg/m2. Erlotinib(150 mg) or gefitinib(250 mg) was administered daily from day 1 to day 21, with a cycle of every 21 days. Dexamethasone, folic acid, and vitamin B12 were also administered during the treatment. The endpoint of the primary study was the disease control rate. Results The objective response rate was 21.9%(95% CI: 7.6% to 36.3%) in the control group, whereas the disease control rate was 84.4%(95% CI: 71.8% to 97.0%) in the experimental group. The median progression-free survival was 6.2(95% CI: 2.4 to 10.0). Grades 3 or 4 adverse effects of leucopenia(15.6%), neutropenia(12.5%), anemia(3.1%), and nausea or vomiting(3.1%) were found in the experimental *** The administration of pemetrexed combined with erlotinib or gefitinib showed a higher efficacy in TKI-resistant NSCLC patients. Further, the adverse effects of this drug combination were well tolerated by the patients. Pemetrexed combined with TKI treatment might provide a satisfactory therapeutic strategy for advanced NSCLC patients after TKI treatment.

关键词：酪氨酸激酶抑制剂非小细胞肺癌疗效观察治疗晚期表皮生长因子受体维生素B12 可信区间

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针对化工系统的分析模型改进

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清华大学学报（自然科学版） 2006年第8期46卷 1473-1477页

作者：彭振宇张冠忠清华大学航天航空学院北京100084

为了确定化工系统中优化改造部件的顺序,针对化工系统的实际特点,提出了建立整体系统目的效率同子系统目的效率关系的数学模型的方法,并且提出了整体效率影响因子的概念作为用能系统优化改造的指导原则。同时以热法磷酸生产系统为例进... 详细信息

为了确定化工系统中优化改造部件的顺序,针对化工系统的实际特点,提出了建立整体系统目的效率同子系统目的效率关系的数学模型的方法,并且提出了整体效率影响因子的概念作为用能系统优化改造的指导原则。同时以热法磷酸生产系统为例进行了建模分析,计算得到了分析的关系模型和各个设备的整体效率影响因子,指出了系统中用能的薄弱环节和改造的重点方向在燃烧塔和水化塔,并同单个子系统目的效率及流图得到的结果和实际改造的结果进行了比较,结果显示,该文方法能够准确方便地指出化工系统能量利用合理性改造的重点方向。

关键词： (火用)分析化工系统模型影响因子

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Influence of tumor response on the survival of patients with extensive-stage small-cell lung cancer treated with the etoposide plus cisplatin chemotherapy regimen

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The Chinese-German Journal of Clinical Oncology 2015年第2期14卷 65-68页

作者： Guojing zhang Yongye Liu Chao Lin Jianfei Guo Long Xu Junling Liu Ying Piao guanzhong zhang Yuhui Liu Yaling Han Xiaodong Xie Department of Oncology PLA Cancer CenterThe General Hospital of Shenyang Military Region Department of Gynaecology The Wenzhou Traditional Chinese Medicine Hospital of Zhejiang Chinese Medical University Department of Cardiology Institute of Cardiovascular Research of PLAThe General Hospital of Shenyang Military Region

Objective In this study, we evaluated the difference of progression-free survival(PFS) and overall survival(OS) between extensive-stage small-cell lung cancer(ES-SCLC) patients who acquired partial response(PR) or complete remission(CR) after two cycles of first-line chemotherapy with the etoposide plus cisplatin(EP) regimen and those who acquired PR or CR after four or six *** A total of 106 eligible patients treated with the EP chemotherapy regimen for two to six cycles, at The General Hospital of Shenyang Military Region(China) between November 2004 and May 2011, were enrolled in this study. RECIST version 1.1 was used for the evaluation of chemotherapy efficiency. We followed up all eligible patients every 4 weeks. All statistical data were analyzed by using SPSS 21.0 statistical package for *** After a median follow-up of 293 days(range, 62–1531 days), all patients had died by the cutoff date. Fifty-one patients acquired PR or CR after two cycles of chemotherapy; the median PFS reached 6.0 months(95% CI, 5.1–6.9), and the median OS was 10.5 months(95% CI, 8.6–12.4). Twenty-eight patients acquired PR or CR after four or six cycles; the median PFS was 4.8 months(95% CI, 4.4–5.2), and the median OS was 7.5 months(95% CI, 6.8–8.2). Both PFS and OS showed a statistical difference between the two groups. Conclusion ES-SCLC patients who acquired PR or CR after two cycles of the EP regimen as first-line therapy had longer PFS and OS than those who acquired PR or CR after four or six cycles.

关键词：患者化疗治疗顺铂肺癌细胞 Windows 反应

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Effects of antioxidant‑rich Lactiplantibacillus plantarum inoculated alfalfa silage on rumen fermentation, antioxidant and immunity status, and mammary gland gene expression in dairy goats

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Journal of Animal Science and Biotechnology 2024年第3期15卷 1227-1240页

作者： Yixin zhang Samaila Usman Qiang Li Fuhou Li Xia zhang Luiz Gustavo Nussio Xusheng Guo State Key Laboratory of Grassland Agro‑ecosystems School of Life SciencesLanzhou UniversityLanzhou 730000PR China Probiotics and Life Health Institute Lanzhou UniversityLanzhou 730000PR China Probiotics and Biological Feed Research Centre Lanzhou UniversityLanzhou 730000PR China Department of Animal Science Luiz de Queiroz College of AgricultureUniversity of Sao PauloPiracicaba 13418‑900Brazil

Background Milk synthesis in lactating animals demands high energy metabolism,which results in an increased production of reactive oxygen metabolites(ROM)causing an imbalance between oxidants and antioxidants thereby inducing oxidative stress(OS)on the *** mitigate OS and postpartum disorders in dairy goats and gain insight into the impact of dietary choices on redox status during lactation,a feeding trial was conducted using alfalfa silage inoculated with a high-antioxidant strain of Lactiplantibacillus *** Twenty-four guanzhong dairy goats(38.1±1.20 kg)were randomly assigned to two dietary treatments:one containing silage inoculated with *** MTD/1(RSMTD-1),and the other containing silage inoculated with high antioxidant activity *** 24-7(ES24-7).Results ES24-7-inoculated silage exhibited better fermentation quality and antioxidant activity compared to *** ES24-7 diet elevated the total antioxidant capacity(T-AOC),superoxide dismutase(SOD),glutathione peroxi-dase(GSH-Px),and catalase(CAT)activities in milk,serum,and feces of lactating goats(with the exception of T-AOC in milk).Additionally,the diet containing ES24-7 inoculated silage enhanced casein yield,milk free fatty acid(FFA)content,and vitamin A level in the goats’***,an increase of immunoglobulin(Ig)A,IgG,IgM,inter-leukin(IL)-4,and IL-10 concentrations were observed,coupled with a reduction in IL-1β,IL-2,IL-6,interferon(IFN)-γ,and tumor necrosis factor(TNF)-αconcentrations in the serum of lactating goats fed *** concentrations of total volatile fatty acid(VFA),acetate,and propionate were observed in the rumen fluid of dairy goats fed ES24-7 inoculated ***,the diet containing ES24-7 inoculated silage significantly upregulated the expression of nuclear factor erythroid 2 like 2(NFE2L2),beta-carotene oxygenase 1(BCO1),SOD1,SOD2,SOD3,GPX2,CAT,glu-tathione-disulfide reductase(GSR),and heme oxygenase 1(HMOX1)genes in the mammary gland,

关键词： Alfalfa silage Antioxidant activity Gene expression Goats Immunity Lactation

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盐酸羟考酮控释片治疗晚期癌症疼痛的临床应用

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中国肿瘤临床 2007年第2期34卷 72-74页

作者：张冠中谢晓冬刘大为沈阳军区总医院肿瘤科沈阳市11000

目的:观察盐酸羟考酮控释片(奥施康定)治疗晚期癌症中、重度疼痛的临床效果、不良反应及患者生活质量的改善情况。方法:68例中、重度疼痛患者给予奥施康定镇痛治疗,初始剂量5mg/12h,正在用吗啡类镇痛药者,按照吗啡1/2剂量换算,根据疼痛... 详细信息

目的:观察盐酸羟考酮控释片(奥施康定)治疗晚期癌症中、重度疼痛的临床效果、不良反应及患者生活质量的改善情况。方法:68例中、重度疼痛患者给予奥施康定镇痛治疗,初始剂量5mg/12h,正在用吗啡类镇痛药者,按照吗啡1/2剂量换算,根据疼痛情况调整剂量,直至患者无痛或基本无痛,每位患者至少治疗15d以上,同时进行疼痛强度、生活质量评分及不良反应观察。结果奥施康定的最终滴定剂量为:≤30mg/d的30例(44.1%),31-60mg/d的16例(23.5%),61-120mg/d的18例(26.5%),≥120mg/d的4例(5.9%)。总疼痛缓解率为95.6%,其中完全缓解27例(39.7%),明显缓解33例(48.5%),中度缓解5例(7.4%)。患者生活质量明显提高,不良反应少且轻微。结论:盐酸羟考酮控释片治疗中、重度癌性疼痛疗效显著,不良反应较少,能显著改善癌症患者的生活质量。

关键词：癌症疼痛盐酸羟考酮控释片治疗

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协商与对抗:争议性事件中的话语解读——基于“东莞扫黄”事件报道与网络文本的量化分析

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新媒体与社会 2014年第4期11卷 259-270页

作者：张冠中安徽大学新闻传播学院

“东莞扫黄”事件本是一个地区性城市治理事件，但因为各类媒介的参与、报道成为全民热议的话题，其中，“东莞挺住”话题一度高居微博话题热议榜榜首。本文采用内容分析法，对代表性官方媒体、专业媒体和民间自媒体分别进行抽样分析，... 详细信息

“东莞扫黄”事件本是一个地区性城市治理事件，但因为各类媒介的参与、报道成为全民热议的话题，其中，“东莞挺住”话题一度高居微博话题热议榜榜首。本文采用内容分析法，对代表性官方媒体、专业媒体和民间自媒体分别进行抽样分析，探究三个舆论场对于这一争议性事件采取的不同的报道与评论框架，以及不同框架之下媒介关注侧重点、报道评论立场等方面的不同。研究发现：民间话语对官方话语进行了戏谑与对抗性解读，而专业媒体则保持了相对独立的声音。

关键词： “东莞扫黄” 框架理论话语博弈舆论场

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螺杆钢球副夹紧机构的设计与研究

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机械传动 2010年第10期34卷 5-7页

作者：张冠中王洪欣徐州建筑职业技术学院机电工程系江苏徐州221116 中国矿业大学机电工程学院江苏徐州221116

专用夹具是实现工件精确定位与快速安装的有效手段,设计出了一种螺旋副与钢球副组合的专用夹紧机构;充分利用了螺旋副的自锁性、钢球副的大增力性与机械效率高的特点,研究了它的输出位移量、力传递的关系与机械效率;通过对设计实例的分... 详细信息

专用夹具是实现工件精确定位与快速安装的有效手段,设计出了一种螺旋副与钢球副组合的专用夹紧机构;充分利用了螺旋副的自锁性、钢球副的大增力性与机械效率高的特点,研究了它的输出位移量、力传递的关系与机械效率;通过对设计实例的分析,展示了夹紧位移、夹紧行程中各运动副中的受力与机械效率的大小。

关键词：螺旋副钢球副夹紧机构设计研究

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