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蛇足石杉中石杉碱甲合成相关基因HsPKS1的克隆及表达分析

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中草药 2024年第18期55卷 6324-6334页

作者：沈玉吴昊吴德财姚诗晴李欣媛戴亮芳罗向东江西师范大学生命科学学院江西南昌330022

目的克隆蛇足石杉Huperzia serrata中石杉碱甲合成相关基因HsPKS1,解析其功能,为揭示石杉碱甲生物合成机制奠定基础。方法基于蛇足石杉转录组及qRT-PCR,筛选并克隆HsPKS1基因,利用生物学信息分析预测其功能;采用同源重组方法构建基因表... 详细信息

目的克隆蛇足石杉Huperzia serrata中石杉碱甲合成相关基因HsPKS1,解析其功能,为揭示石杉碱甲生物合成机制奠定基础。方法基于蛇足石杉转录组及qRT-PCR,筛选并克隆HsPKS1基因,利用生物学信息分析预测其功能;采用同源重组方法构建基因表达载体,遗传转化至烟草中进行亚细胞定位和表达分析,利用广泛靶向代谢组分析HsPKS1基因参与的生物合成通路的作用机制。结果qRT-PCR和HPLC结果表明不同组织间HsPKS1基因相对表达量与其石杉碱甲含量呈正相关。基因克隆、测序及生物信息学分析表明,HsPKS1基因开放阅读框为1200 bp,编码399个氨基酸,编码蛋白具有酰基转移酶活性区域及多个辅酶A的结合位点。系统进化树分析表明,HsPKS1蛋白与25个物种的聚酮合酶均具有较高的同源性,其中与江南卷柏的亲缘关系最近。亚细胞定位结果表明HsPKS1蛋白在细胞核和细胞质均有表达。广泛靶向代谢组分析发现,HsPKS1基因可以显著影响烟草次生代谢产物的积累,并引起合成黄酮类的前体化合物和异喹啉类生物碱化合物的物质变化。结论成功克隆并获得HsPKS1基因序列,分析了基因表达模式、生物学功能及亚细胞定位,揭示了该基因在类黄酮和生物碱合成等方面起重要作用,为研究和揭示HupA合成代谢通路的分子机制提供科学依据。

关键词：蛇足石杉聚酮合酶基因克隆烟草瞬时表达广泛靶向代谢组

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运动时间对结肠镜检查患者肠道清洁度的影响

分子影像学杂志 2017年第4期40卷 504-507页

作者：宋希殷小雅王华军戴世学李良芳张华娟广东省医学科学院//广东省人民医院消化内科广东广州510010 南方医科大学公共卫生学院广东广州510515

目的探讨不同的运动时间对于提高药物清洁肠道作用的效果,从而优化肠道清洁方案,提高结肠镜检查准确性。方法收集2017年2月~2017年9月广东省人民医院消化一区需要进行结肠镜检查的住院患者作为观察对象,共入选60例患者,其中男性41例,女... 详细信息

目的探讨不同的运动时间对于提高药物清洁肠道作用的效果,从而优化肠道清洁方案,提高结肠镜检查准确性。方法收集2017年2月~2017年9月广东省人民医院消化一区需要进行结肠镜检查的住院患者作为观察对象,共入选60例患者,其中男性41例,女性19例,年龄25~83岁;将以上患者随机分为A、B、C组,每组20例,3组的性别构成及年龄差异均无统计学意义(P>0.05)。采用国际公认的波士顿Boston肠道准备量表评分,比较3组患者的肠道清洁度;并比较分析3组的泻药不良反应、对运动的耐受性。结果 3组患者肠段清洁度差异有统计学意义(P<0.001);对泻药的不良反应差异无统计学意义(P=0.067);C组对运动的耐受性最差(P<0.05)。结论口服复方聚乙二醇后散步30~60 min是较为理想、有效的肠道准备方法,可满足结肠镜检查的需要。

关键词：时间结肠镜检查肠道准备肠道清洁度运动散步

The transcription factor KLF14 regulates macrophage glycolysis and immune function by inhibiting HK2 in sepsis

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Cellular & Molecular Immunology 2022年第4期19卷 504-515页

作者： Yuan Yuan Guangjian Fan Yuqi liu Lu liu Tong Zhang Pengfei liu Qing Tu Xinyi Zhang Shiyuan Luo liangfang Yao Feng Chen Jingbao li Department of Anesthesiology Shanghai General HospitalShanghai Jiao Tong UniversityShanghai201620China Precision Research Center for Refractory Diseases Shanghai General HospitalShanghai Jiao Tong UniversityShanghai201620China Department of Anesthesiology Weifang Medical UniversityWeifang261000China Department of Anesthesiology Ruijin HospitalShanghai Jiao Tong University School of MedicineShanghai201620China

Sepsis is a heterogeneous syndrome induced by a dysregulated host response to *** plays a role in maintaining the immune function of macrophages,which is crucial for severely septic ***,how the pathways that link glycolysis and macrophages are regulated is still largely ***,we provide evidence to support the function of KLF14,a novel Krüppel-like transcription factor,in the regulation of glycolysis and the immune function of macrophages during ***14 deletion led to significantly increased mortality in lethal models of murine endotoxemia and ***,KLF14 decreased glycolysis and the secretion of inflammatory cytokines by macrophages by inhibiting the transcription of *** addition,we confirmed that the expression of KLF14 was upregulated in septic ***,pharmacological activation of KLF14 conferred protection against sepsis in *** findings uncover a key role of KLF14 in modulating the inflammatory signaling pathway and shed light on the development of KLF14-targeted therapeutics for sepsis.

关键词： Sepsis Macrophages KLF14 HK2 Glycolysis Perhexiline

Fiber-modified adenoviral vector expressing the tumor necrosis factor-related apoptosis-inducing ligand gene from the human telomerase reverse transcriptase promoter induces apoptosis in human hepatocellular carcinoma cells

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World Journal of Gastroenterology 2005年第17期11卷 2552-2556页

作者： DietmarJacob GuidoSchumacher MarcusBahra JohnDavis Hong-BoZhu li-DongZhang FuminoriTeraishi PeterNeuhaus Bing-liangfang DepartmentofGeneral VisceralandTransplantationSurgeryHumboldtUniversityofBerlinCharitéVirchowClinicBerlinGermany DepartmentofThoracicandCardiovascularSurgery TheUniversityofTexasMDAndersonCancerCenter DepartmentofThoracicandCardiovascularSurgery TheUniversityofTexasMDAndersonCancerCenter//PrograminGeneTherapyandVirologyTheUniversityofTexasGraduateSchoolofBiomedicalSciencesHoustonTX77030USA

AIM: Because of a major resistance to chemotherapy, prognosis of hepatocellular carcinoma (HCC) is still poor. New treatments are required and gene therapy may be an option. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in multiple malignant tumors, and using adenoviral vectors has shown a targeted tumor-specific therapy. However, repeated administration of adenoviral vectors can lead to cell resistance, which may be caused by the initial coxsackie-adenovirus receptor (CAR). One technique to overcome resistance is the use of modified adenoviral vectors containing an Arg-Gly-Asp (RGD) sequence. In this study we constructed an adenoviral vector (designated Ad/TRAIL-F/RGD) with RGD-modified fibers, expressing the TRAIL gene from the human telomerase reverse transcriptase (hTERT) promoter, and evaluated its antitumor activity in HCC cell ***: To investigate the effects of Ad/TRAIL-F/RGD in human HCC cell lines Hep G2 and Hep 3b, cells were infected with Ad/CMV-GFP (vector control), Ad/gTRAIL (positive control), and Ad/TRAIL-F/RGD. Phosphatebuffered saline (PBS) was used as control. Cell viability was determined by proliferation assay (XTT), and apoptosis induction by fluorescence activated cell sorting (FACS).RESULTS: Cells treated with Ad/TRAIL-F/RGD and Ad/ gTRAIL showed a significantly reduced cell viability in comparison to PBS and Ad/CMV-GFP treatment in both cell lines. Whereas, treatment with PBS and Ad/CMVGFP had no cell-killing effect. The reduced cell viability was caused by induction of apoptosis as shown by FACS analysis. The amount of apoptotic cells was similar after incubation with Ad/gTRAIL and Ad/TRAIL-F/RGD. CONCLUSION: The new RGD modified vector Ad/TRAILF/RGD could become a potent therapeutic agent for the treatment of HCC, adenovirus resistant tumors, and CAR low or negative cancer cells.

关键词： HCC TRAIL hTERT

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我国青贮玉米栽培技术研究进展

黑龙江农业科学 2024年第9期 114-119页

作者：杨克泽吴之涛李文学常浩汪亮芳徐志鹏杨小龙刘强甘肃省农业工程技术研究院/武威市玉米病虫害绿色防控技术创新中心/甘肃省玉米病虫害绿色防控工程研究中心甘肃武威733000

近年来,随着我国畜牧业的发展壮大和种植业结构调整,青贮玉米产业发展突飞猛进,对我国国民经济发展具有重要意义。随着“粮改饲”政策的实施,我国青贮玉米种植面积快速增加。结合前人最新研究进展和本团队工作实际,从播期、种植密度、... 详细信息

近年来,随着我国畜牧业的发展壮大和种植业结构调整,青贮玉米产业发展突飞猛进,对我国国民经济发展具有重要意义。随着“粮改饲”政策的实施,我国青贮玉米种植面积快速增加。结合前人最新研究进展和本团队工作实际,从播期、种植密度、施肥、种植模式、覆膜和收获时期等方面对青贮玉米生长及产量的影响研究进行综述和展望,今后需要对青贮玉米栽培方式进行系统研究和优化,明确各生产要素与青贮玉米产量和品质间的关系,创新集成绿色高效生产技术,积极应用和创新种植模式,实现技术研究与生产应用的结合。针对不同品种、地域和增产手段进行研究比较,重点研究可适用于大规模推行的绿色高效种植技术,破解青贮玉米高效种植关键技术和推广难题;加大科研投入,通过项目实施积极搭建产学研交流平台和技术推广体系,培养青贮玉米科技人才;积极制定不同生态区适应栽培品种技术规范,加大青贮玉米品种的推广示范力度,加强栽培技术的培训和宣传;做好病虫害的绿色防控,显著增加青贮玉米营养价值、生物产量和质量,从而指导青贮玉米产业快速高效发展。

关键词：青贮玉米栽培技术产业发展

Expression pattern of epithelial cell adhesion molecule on normal and malignant colon tissues

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World Journal of Gastroenterology 2005年第3期11卷 344-347页

作者： XinXie Chun-YanWang Yun-XinCao WeiWang RanZhuang li-HuaChen Na-NaDang liangfang Bo-QuanJin DepartmentofImmunology FourthMilitaryMedicalUniversityXi'an710032ShaanxiProvinceChina DepartmentofImmunology FourthMilitaryMedicalUniversityXi'an710032ShaanxiProvinceChina

AEM: To investigate the expression pattern of epithelial cell adhesion molecule (Ep-CAM) on normal and malignant colon tissues to evaluate its diagnostic and therapeutic significance. METHODS: cDNA encoding Ep-CAM extracellular domain was doned by reverse transcription-polymerase chain reaction (RT-PCR) from excised malignant colon tissues and inserted into a glutathione S-transferase (GST)-tagged vector. Ep-CAM-GST fusion protein was induced by isopropyl-p-D-thiogalactopyranoside (IPTG) and purified with glutathione-sepharose. The Ep-CAM-GST fusion protein was mixed with Freund's adjuvant and Balb/c mice were immunized with it. Sp2/0 myeloma cells were fused with the spleen cells of the immunized mice. After having selected by indirect EliSA, the anti-Ep-CAM monoclonal antibodies (MAbs) were generaled and the corresponding ascites were obtained. Finally, the human colon carcinoma tissue array prepared from seventy individual patients was stained with the anti-Ep-CAM MAbs. RESULTS: The isdated Ep-CAM cDNA sequence was identical to the data in GenBank. The expressed fusion protein was almost soluble and had a molecular weight (MW) of 53 ku. Four MAbs against Ep-CAM were obtained and designated as FMU-Epl, FMU-Ep2, FMU-Ep3 and FMU-Ep4 respectively. Among them, FMU-Ep4 could recognize the natural Ep-CAM on Colo205 and SW480 cells, and all of them could be used for immunohistochemical staining of tissue sections. It was fbund that Ep-CAM was distributed differently in normal and various malignant colon tissues, induding squamous cell carcinoma, signet-ring cell carcinoma and adenocarcinoma. In normal colon gland epithelia, Ep-CAM antigen was mainly distributed on the basolateral membrane and in the region between the basolateral membrane and the cytoplastic part near the nuclei, whereas the expression pattern of colon malignancies was mainly on the whole surface of epithelia and the expression was much higher than the normal colon tissues. The staining pattern of tissue

关键词： Coon carcinoma Coon Epithelial cell adhesion molecule

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我国青贮玉米研究进展及发展对策

寒旱农业科学 2024年第9期3卷 796-803页

作者：杨克泽王丽梅汪亮芳吴之涛李文学常浩徐志鹏杨小龙刘强牛小霞甘肃省农业工程技术研究院甘肃兰州730030 武威市玉米病虫害绿色防控技术创新中心甘肃武威733000 甘肃省玉米病虫害绿色防控工程研究中心甘肃兰州730030 武威市凉州区农业技术推广中心甘肃武威733000 甘肃省农业科学院经济作物与啤酒原料研究所甘肃兰州730070

青贮玉米是青贮饲料的重要组成部分,发展青贮玉米是促进我国畜牧业高质量发展的重要举措。大量的青贮玉米研究已取得了一系列的重要进展,结合前人最新研究进展和工作实际,对我国青贮玉米育种、品种适应性、栽培技术、病虫害防治、机械... 详细信息

青贮玉米是青贮饲料的重要组成部分,发展青贮玉米是促进我国畜牧业高质量发展的重要举措。大量的青贮玉米研究已取得了一系列的重要进展,结合前人最新研究进展和工作实际,对我国青贮玉米育种、品种适应性、栽培技术、病虫害防治、机械化水平和青贮技术等方面进行了概述,并对我国青贮玉米发展面临的主要问题进行了分析,提出了相应的发展对策,即加强科技创新,搭建现代青贮玉米规模化育种体系平台;扩大生产规模,提高青贮玉米全程机械化配套效率;加强政策扶持力度,加大资金投入和科技人才培养;促进产学研结合,积极搭建青贮玉米标准化生产技术推广体系。旨在为我国青贮玉米产业高质量发展提供一定的参考。

关键词：青贮玉米育种现状病虫害研究发展对策

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分级阅读推广的尴尬与出路

出版发行研究 2010年第10期 15-18页

作者：吴亮芳李建红武汉大学信息管理学院中国书籍出版社

儿童阅读及接受能力与年龄、心智发展整体上呈吻合态势,为分级阅读提供了理论依据。我国儿童图书出版市场的无序化与儿童阅读水平不容乐观的状况急需进行分级阅读推广,而推广中却遭遇了种种尴尬,诸如推广主体单一、推广行为遭质疑、推... 详细信息

儿童阅读及接受能力与年龄、心智发展整体上呈吻合态势,为分级阅读提供了理论依据。我国儿童图书出版市场的无序化与儿童阅读水平不容乐观的状况急需进行分级阅读推广,而推广中却遭遇了种种尴尬,诸如推广主体单一、推广行为遭质疑、推广信息不对称。这些遭遇,从侧面也给我们指明了出路,如分级阅读的推广需要营造一种氛围、需要突出它的公益性、需要一个权威的分级标准,更需要多方的共同努力,最终实现全民参与分级阅读的推广。

关键词：分级阅读推广必要性尴尬出路

REGγ Mediated Regulation of p21Waf/Cip1, p16INK4aand p14ARF/p19ARFin Vivo

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Journal of Cancer Therapy 2013年第5期4卷 933-938页

作者： Lei li Pei Zhang Haibin Wei Weicang Wang liangfang Yao Yubing li Lei Zhou Zhuo Wang Jiang liu li Zhou Ali Amjad Bianhong Zhang Tiantian Jing Jianru Xiao Yongyan Dang Xiaotao li Department of Orthopaedic Oncology Changzheng Hospital The Second Military Medical University Shanghai China Department of Pathology The Second Chengdu Municipal Hospital Chengdu China The Shanghai Key Laboratory of Regulatory Biology Institute of Biomedical Sciences East China Normal University Shanghai China

p21Waf/Cip1, p16INK4a and p14ARF (p19ARF in mice) have been demonstrated to be degraded by REGγ-proteasome pathway in an ATP- and ubiquitin-independent manner in vitro. However, the in vivo roles of REGγ mediated-degradation of p21Waf/Cip1, p16INK4a and p14ARF remain unclear. In this study, we showed enhanced expression of p21Waf/Cip1, p16INK4a and p19ARF in multiple tissues from REGg–/– mice compared to REGg+/+ mice. Furthermore, we examined the expression of p21Waf/Cip1, p16INK4a and p14ARF in different cancer tissues and observed that the REGγ protein levels were highly expressed in different human cancers while the level of p21Waf/Cip1, p16INK4a and p14ARF appears to be inversely correlated. These results demonstrate that REGγ may exert its function in physiological and pathological conditions through degradation of p21Waf/Cip1, p16INK4a and p14ARF in vivo.

关键词： REGγ p21Waf/Cip1 p16INK4a p14ARF p19ARF Cancer