运用融合PCR技术敲除光滑念珠菌FLO8基因以及其对EPA黏附素家族表达的影响

目的:构建光滑念珠菌FLO8基因敲除株,并分析FLO8敲除对光滑念珠菌上皮黏附素(epithelial adhe-sin,EPA)家族表达的影响。方法:使用融合PCR技术,以光滑念珠菌ATCC2001菌株基因组DNA、带有筛选标记诺尔斯菌素抗性基因(NAT)的质粒DNA为模板,构建敲除组件。采用醋酸锂转染法将敲除组件转染入ATCC2001中,从而获得flo8△菌株。使用实时荧光定量PCR检测菌株中EPA1、EPA6和EPA7基因的表达。结果:获得光滑念珠菌FLO8基因敲除株flo8△,该菌株的EPA1、EPA6和EPA7基因表达水平对明显低于ATCC2001株(P均<0.001)。结论:此法可便捷、有效地构建光滑念珠菌基因敲除菌株。敲除FLO8基因后,光滑念珠菌的EPA家族表达降低,为进一步研究光滑念珠菌毒力机制奠定基础。

基于FLO-2D的抽水蓄能电站泥石流模拟分析

针对新疆吐鲁番盆地东侧鄯善县泥石流灾害影响该地区工程建设与发展的问题,以新疆鄯善县抽水蓄能电站下库区泥石流为研究对象,运用FLO-2D数值模拟软件,对不同降雨频率下(P=1%,2%,5%)泥石流沟的发生过程进行模拟。结果表明:在降雨频率P=1%和P=2%的降水条件下,沟内泥石流将冲出淤积在拦沙坝附近,并侵占河道。以库区运行年限100 a计,库区淤积总量为21567 m^(3),对下库区库容产生一定的影响。

Mechanism of Rosae Rugosae Flos flavonoids in the treatment of hyperlipidemia and optimization of extraction process based on network pharmacology

This study aims to identify a natural plant chemical with hypolipidemic effects that can be used to treat high cholesterol without adverse reactions.Through network pharmacology screening,it was found that Rosae Rugosae Flos(RF)flavonoids had potential therapeutic effects on hyperlipidemia and its mechanism of action was discussed.TCMSP and GeneCards databases were used to obtain active ingredients and disease targets.Venn diagrams were drawn to illustrate the findings.The interaction network diagram was created using Cytoscape 3.8.0 software.The PPI protein network was constructed using String.GO and KEGG enrichment analysis was performed using Metascape.The results revealed 2 active flavonoid ingredients and 60 potential targets in RF.The key targets,including CCL2,PPARG,and PPARA,were found to play a role in multiple pathways such as the AGE-RAGE signaling pathway,lipid and atherosclerosis,and cancer pathway in diabetic complications.The solvent extraction method was optimized for efficient flavonoid extraction based on network pharmacology prediction results.This was achieved through a single factor and orthogonal test,resulting in an optimum process with a reflux time of 1.5 h,a solid-liquid ratio of 1:13 g/mL,and an ethanol concentration of 50%.

Optimization of extraction process for total flavonoids of Sophorae Flos for the treatment of hyperlipidemia based on network pharmacology and molecular docking

This study aimed to investigate the mechanism of action of Sophora Flos(SF)in the treatment of hyperlipidemia(HLP)using network pharmacology and molecular docking methods,and to optimize the extraction process of the predicted active components.The STRING database was used for protein interaction analysis and PPI network construction via Cytoscape 3.9.1.Pymol was employed for docking and visualization.An extensive review of SF identifi ed 6 active ingredients,297 related objectives,84 disease objectives,and 57 total objectives.After protein interaction and topology analysis,18 core targets were identified.These included 146 gene function entries(P<0.05).Active compounds,mainly flavonoids,can modulate the expression of various proteins such as TNF,IL-6,IL-1β,PPARG,and TGFB1 to achieve therapeutic effects on HLP.The network pharmacology and molecular docking results suggested that the active fl avonoids component in SF may be related to the treatment of hyperlipidemia.Therefore,the orthogonal experiment method was used to optimize the extraction process of total fl avonoid from SF using ethanol refl ux extraction,based on a single factor experiment.The effects of refl ux time,solid-liquid ratio,ethanol concentration,and other factors on the extraction of total fl avonoid from SF were investigated.The optimum process conditions were refl ux time of 1.25 h,solid-liquid ratio of 1:15 g/mL and ethanol concentration of 60%.Using these conditions,the purity of total fl avonoid extracted from SF was 70.33±0.22%.

基于FLO-2D数值模拟的不同降雨频率工况下八宝镇拱北槽沟泥石流运动特征及危险性评价

近年来,随着降雨量不断增加,泥石流爆发频率呈上升趋势,其运动堆积特性及防治工程措施直接决定泥石流危害程度及影响范围,也是影响泥石流危险度区划的关键因素。因此,对不同工况条件下泥石流堆积特征及危险范围预测具有较强的现实应用价值。FLO-2D作为一款专用于洪水和泥石流模拟的二维软件,能较完整地分析泥石流的运动状态特征,可用于模拟洪水灾害和风险评估,推算流体运动的流速及堆积深度,合理推估淹没面积。该文以拱北槽沟泥石流为例,通过开展野外详细调查,在查明泥石流物源类型、分布特征、物源储量与流域育灾环境的基础上,模拟分析其在不同降雨频率下(10%、5%、2%和1%)的泥石流运动特征,进而确定泥石流灾害威胁范围、堆积深度、流速等特征参数,并在此基础上采用泥石流强度指标对泥石流灾害进行危险性区划评价,为拱北槽沟泥石流的监测预警与风险防控提供技术依据,以此为该地区或者其他祁连山区城镇泥石流灾害防治及国土空间规划提供科学依据及参考建议。

Structural characteristics,anticoagulant and antithrombotic mechanism of a novel polysaccharide from Rosa Chinensis Flos

This is the first report on a polysaccharide(RCJ2-Ib)isolated from Rosa Chinensis Flos.RCJ2-Ib was obtained through the extraction with water,precipitation with ethanol,separation with DEAE-52 column and purification with DEAE-Sepharose Fast Flow column and Sephadex G100 column.GC,FT-IR and NMR analyses revealed that RCJ2-Ib(3.3 k Da)was a 1,4-linked polymannuronic acid containing substantialβ-Danomers units.The anticoagulant effect of RCJ2-Ib evaluated by using rabbit ear venous blood and an acute blood stasis rat model showed that RCJ2-Ib had obvious anticoagulant activity in regulating endogenous and exogenous coagulation pathways and reducing serum thromboxane B2 and endothelin-1.In addition,RCJ2-Ib could also increase the number of Lactobacillus and Escherichia coli.As a result,RCJ2-Ib has the potential to inhibit thrombosis and maintain the intestinal environment.

Screening hepatoprotective effective components of Lonicerae japonica Flos based on the spectrum-effect relationship and its mechanism exploring

Lonicerae japonicae Flos(LF)is a kind of healthcare food with hepatoprotective function.This study was designed to explore the spectrum-effect relationships between UPLC fingerprints and the hepatoprotective effects of LF.Fingerprints of ten batches of LF were established by UPLC-PDA.The inhibitory levels of AST and ALT were used as pharmacological indexes,and secoxyloganin,isochlorogenic acid A and isochlorogenic acid C were screened as hepatoprotective active compounds by grey relational analysis(GRA)and partial least squares regression analysis(PLSR).Caspase-3 was obtained by network pharmacology as a key target of hepatoprotective active compounds.Molecular docking is used to explore the interaction between small molecules and proteins.This work provided a general model of the combination of UPLC-PDA and hepatoprotective effect to study the spectrum-effect relationship of LF,which can be used to considerable methods and insight for the fundamental research of the material basis of similar healthcare food.

Chemical compositions and biological activities of Flos lonicerae

Flos lonicerae is the dry flower bud of Lonicera japonica Thunb,and it is a very important Chinese herbal medicine in China.In ancient times,it was widely used to treat the exopathogenic wind-heat,epidemic febrile diseases,sores,carbuncles and some infectious diseases.Modern pharmacological research has indicated that it has anti-inflammatory,anti-bacterial,anti-viral and anti-oxidation activities.This paper reviews the chemical compositions and biological activities of Flos lonicerae in order to provide reference for its application in clinical practice in the future.

基于FLO-2D的四川小金县炭厂沟危险性评价

【目的】研究炭厂沟泥石流爆发时的危险性。【方法】以炭厂沟1:5 000等高线、50年降雨数据和掩膜作为基础,结合现场勘察情况,运用FLO-2D和GIS软件,模拟炭厂沟在不同降雨频率下(10%、5%、2%和1%)泥石流的运动堆积过程。【结果】以1%降雨频率为例,模拟出的最大泥石流流速为20.97 m/s,最大泥石流流深为7.7 m,堆积面积为33 175 m^(2)。极端危险区范围占比81.8%,具有极高的危险性。对比模拟计算流速与公式计算流速来验算精确度,结果显示可靠性高。【结论】通过选用流速和泥深作为泥石流危险度分区的定量指标,得到炭厂沟泥石流的危险性分区图,为该流域的灾害防治提供科学的参考依据。

Study on Quality Standard of Zhuang Medicine Buddlejae Flos

[Objectives]This study was conducted to explore the quality control method and establish the quality standard of Zhuang medicine Buddlejae Flos.[Methods]The microscopic identification method was adopted to identify the characters and microscopic characteristics of dried Buddlejae Flos in combination with traditional experience.The moisture,ash,acid-insoluble components and extracts of Buddlejae Flos were determined with reference to general rules of Chinese Pharmacopoeia(2015 edition).TLC identification was performed.Liquid chromatography-mass spectrometry(LC-MS)was used to determine the contents of linarin and verbascoside in Buddlejae Flos,and gas chromatography-mass spectrometry(GC-MS)was adopted to detect pesticide residues in Buddlejae Flos.Heavy metal elements As,Cd and Pb were detected by inductively coupled plasma mass spectrometry(ICP-MS).[Results]The established method is simple and accurate.Clear spots were observed on the thin layer chromatograms,and the resolution was good.The average value of moisture content was 12.24%;the average value of ash content was 5.1%;the average value of acid-insoluble content was 7.5%;and the average value of extract content was 27.3%.The regression equation of rutin in Buddlejae Flos was y=11.896x-0.0049,R^(2)=0.9996,and the contents of linarin and verbascoside were 5680 and 2080 mg/kg,respectively.No pesticide residues and heavy metals were detected in the medicinal materials of Buddlejae Flos.[Conclusions]This study can provide reference for the quality standard control of Guangxi Zhuang medicine Buddlejae Flos.

基于FLO^(2D)与3DMine耦合的尾矿库溃坝灾害模拟

为预测和评价尾矿库溃坝的致灾情况,提高库区防灾减灾能力,建立尾矿库溃坝数值仿真模型。采用FLO2D泥石流模拟软件对湖南某尾矿库进行溃坝灾害过程仿真,并将模拟结果与3DMine数字矿山软件耦合,实时反演溃坝灾害过程。分析结果表明,该尾矿库溃坝时的最大溃决流速为40.2 m/s,下游沟谷最大淹没深度达13.6 m,溃坝瞬时最大冲击应力达100 MPa。三维动态反演了尾矿库溃坝前稳定状态、溃坝后0.01,0.02和0.05 h的尾砂流动状态和淹没范围。根据溃坝泥砂流动强度,对该尾矿库全部溃坝和半溃坝时的灾害程度进行三色图判别。

絮凝基因(FLO1G)的序列测定及分析

The sequence of the flocculation gene (FLO1G) was determined. The result of sequcencing showed that:the cloned gene contains a large open reading frame (ORF) of 3936 bp and encodes for a protein of 1312 amino acid. According to the result of homologous analysis, the cloned gene is homologous to FLO1 but with 675 bp deletion in the ORF region. The missing part belongs to one of the four repeated sequence family of FLO1. Since the cloned DNA fragment can trigger strong flocculence to non-flocculent strain S.cerevisiae YS58, we concluded that the missing part is not the crutical part for the flocculent ability of the gene.

龙眼FLO/LFY同源基因cDNA片段的克隆

根据不同植物FLO LFY同源基因3′端保守序列,设计简并引物,运用RT＿PCR技术在龙眼花芽总RNA中扩增得到长度为408bp的cDNA片段,命名为lonFLO。序列分析表明,该片段与金鱼草FLO、苹果AFL2、AFL1、拟南芥LFY有较高的核苷酸同源性,分别达98 8%,81 6%,81 1%和76 2%。

FLO和FLP方案治疗晚期胃癌的临床观察

目的探讨FLO和FLP方案治疗晚期胃癌的疗效和毒副作用。方法将62例晚期胃癌患者随机分为A、B两组,分别用FLO方案（A组）和FLP方案（B组）化疗,每2周为1个周期,共8个周期,观察其疗效、骨髓毒性、呕吐副作用及外周神经毒性等。结果治疗有效率A组为41%,B组为17%（P〈0.01）;Ⅲ~Ⅳ度骨髓损害发生率A组为38%,B组为37%（P〉0.05）;Ⅲ~Ⅳ度呕吐发生率A组为9%,B组为30%（P〈0.01）;Ⅰ~Ⅱ度外周神经毒副作用发生率A组为72%,B组为7%（P〈0.01）,Ⅲ~Ⅳ度外周神经毒副作用发生率A组为3%,B组为0%（P〉0.05）。结论 FLO较FLP治疗晚期胃癌疗效更好,呕吐更轻,虽然外周神经毒副作用多,但症状轻。

基于FLO-2D模型的泥石流运动冲淤数值模拟

采用2维流变模型FLO-2D,经连续方程与运动方程,结合网格化的数字高程模型,选取陇南市武都地区清水沟、嘴子沟及段河坝3条泥石流沟为试验区.通过DEM在ArcGIS中转换为ASCⅡ格式输入至FLO-2DPRO软件中,建立流域模型,分析计算出各网格点的可能运动速度、淤积深度,并将模拟结果整合致GIS中.在GIS软件中分析可能淤积范围,通过对比2010年陇南"8.12"特大暴雨事件引发泥石流灾害灾情,验证了FLO-2D模型模拟试验区泥石流威胁范围的正确性,发现模拟结果与实际发生灾情威胁范围大致相符.

基于FLO-2D的溃决型泥石流模拟研究

2013年7月10日华溪沟引发降雨量为100年一遇的溃决型泥石流("7.10"泥石流),造成岷江堵塞并形成堰塞湖,造成巨大经济损失和人员伤亡。为了研究溃决型泥石流危险性,探讨不同降雨频率下暴发泥石流的结果,利用FLO-2D软件对华溪沟在不同降雨频率下溃决与非溃决条件进行了数值模拟,并把100年一遇的模拟结果与实际野外调查情况做出对比。结果显示溃决条件下冲出量、堆积范围面积、平均堆积深度比非溃决条件下超出1倍;溃决时冲出量比实际调查值少5.58万m3。数值模拟结果表明模型的误差在允许范围内,模拟结果与原型相比准确度为88.61%,模拟效果较好。溃决型泥石流规模比非溃决时相比甚大,因此在进行灾害防治设计和危险性评价时应特别注意。

基于FLO-2D与冲量模型的泥石流危险度分区方法及应用

鉴于正确的泥石流危险度分区对合理规划布局泥石流堆积区经济活动的重要作用,采用FLO-2D模型对泥石流的运动堆积过程进行数值模拟,求得泥石流堆积扇上流速、流深分布及以格网为单位的泥石流体流速和流深的时间变化过程,结合GIS技术对计算所得数据进行空间分析,以泥石流体冲量为分区指标,建立泥石流堆积扇危险度分区模型,并以四川省石棉县马颈子沟泥石流为例,模拟该沟百年一遇泥石流运动堆积过程。结果表明,模拟堆积范围与实际堆积面积吻合度达86.16%;根据格网内泥石流体冲量模长,可将泥石流堆积扇危险度分为极度、重度、中度、轻度危险区四个等级。

融合PCR结合同源重组技术敲除白色假丝酵母菌FLO8基因

目的运用融合PCR和同源重组技术敲除白色假丝酵母菌FLO8基因,并初步分析敲除菌株药物敏感性(药敏)情况。方法运用融合PCR将FLO8基因的上下游片段与筛选标记融合在一起构成同源敲除组件,再用高效醋酸锂转染法将敲除组件转染入白色假丝酵母菌SN152,在营养缺陷板上进行筛选,通过2次同源重组敲除FLO8的2条等位基因。采用点板法比较FLO8未敲除株与敲除株药敏情况。结果成功构建白色假丝酵母菌SN152 FLO8双等位基因缺失株,且耐药性FLO8^(-/-)>FLO8^(+/-)>SN152。结论融合PCR结合同源重组技术能高效、快速构建白色假丝酵母菌FLO8基因缺失株,白色假丝酵母菌FLO8基因与药敏相关,且FLO8拷贝数越少耐药性越强。

基于Flow-R和FLO-2D的沟谷型泥石流危险性评价

泥石流是中国西南山区频发的地质灾害之一,具有突发性、流速快以及破坏力强等特点,往往造成巨大的人员伤亡和经济损失。地质灾害危险性评估是防灾减灾管理和防治环节中的有效措施之一,用于合理量化泥石流运动堆积线路沿程泥石流灾害危险性的空间分布特征。以四川省都江堰市龙池镇麻柳沟为研究对象,选取坡度、平面曲率和上坡汇水面积作为沟谷型泥石流启动区识别因子,基于Flow-R模型识别出的泥石流启动点,耦合FLO-2D模型模拟麻柳沟泥石流运动堆积过程,将泥石流堆积深度、运动速度和暴发频率相结合分析泥石流的危险性,并采用混淆矩阵对模拟结果进行了精度评估。结果表明:基于Flow-R和FLO-2D耦合模型的麻柳沟泥石流的运动堆积模拟结果与实际情况吻合度较高。模拟泥石流堆积区面积2.19万m^(2),堆积体积4.92万m^(3),泥石流危险性由沟道中心向两侧逐渐降低,高危险区占比42.3%,中危险区占比23.1%,低危险区占比34.6%。麻柳沟形成区、流通区沟道中散落堆积物体积大约为30万m^(3),麻柳沟泥石流仍处于高度活跃阶段,危险性较高,今后在降雨和水动力充分的条件下大量松散物质可能将被重新激活,一旦发生泥石流将有可能造成重大地质灾害。

基于FLO-2D的核桃沟泥石流数值模拟分析

铁路沿线泥石流灾害将对铁路隧道和桥梁施工和运营造成极大危害。文章选取核桃沟为研究对象,运用FLO-2D软件模拟不同周期的降雨过程,并提取降雨过程线计算泥石流洪峰流量。通过与雨洪法计算结果对比,验证了根据降雨过程线确定泥石流洪峰流量的合理性,进而对不同暴雨重现周期下核桃沟支沟和主沟泥石流发生过程进行模拟。核桃沟支沟泥石流模拟结果与分析表明,不同重现周期下,泥石流对支沟2~#斜井、沟谷居民安置点以及工程项目部的威胁较大,建议加强对支沟的泥石流监测预警。核桃沟主沟泥石流模拟结果与分析表明,不同重现周期下,核桃沟大桥处泥石流流速与堆积厚度有增大趋势;在50或100a一遇情况下,泥石流对核桃沟大桥的影响较大。