Ray-tracing polymorphic multidomain spectralhp elements for isosurface rendering
计算机专业用语
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重点实验室获国家自然科学基金资助项目
重点实验室获国家自然科学基金资助项目
佚名
【期刊名称】《今日科技》
【年(卷),期】2012(000)009
【摘要】近日前,接国家自然科学基金委员会通知,重点实验室杨章女博士今年负责申报的题为“莱姆螺旋体保守基因簇bba68-bba73的生物学功能研究”项目获国家青年科学基金项目资助,批准号为81201319。
该项目为该重点实验室继朱函坪主任技师、姚苹苹副主任技师相继在2007年、2011年获国家基金项目资助后的第3个主持的资助项目。
【总页数】1页(P57-57)
【正文语种】中文
【中图分类】S88-55
【相关文献】
1.我院一科技项目获国家自然科学基金项目资助 [J], ;
2.复旦医科461个项目获2019年度国家自然科学基金项目立项资助 [J], 无
3.我校6项国家自然科学基金项目、3项国家社科基金项目获资助立项 [J],
4.2006年度国家自然科学基金资助项目及项目负责人专栏介绍华中科技大学服务计算技术与系统教育部重点实验室——陶文兵 [J],
5.我校8个项目获2013年度国家自然科学基金项目立项资助 [J],
因版权原因,仅展示原文概要,查看原文内容请购买。
mTORC1
mTORC1/2双重抑制剂OSI -027抑制高氧诱导的肺成纤维细胞增殖和分化*吴黎虹, 唐坤, 党红星△, 符跃强, 刘成军, 李静, 许峰(重庆医科大学附属儿童医院重症医学科,国家儿童健康与疾病临床医学研究中心,儿童发育疾病研究教育部重点实验室,儿科学重庆市重点实验室,重庆 400014)[摘要] 目的:分析哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin , mTOR )复合物1/2(mTORcomplex 1/2, mTORC1/2)双重抑制剂OSI -027对高体积分数氧(高氧)所致人胚肺成纤维细胞增殖和分化的抑制作用。
方法:高氧(95% O 2)处理人胚肺成纤维细胞MRC -5建立增殖分化模型,分为对照组、高氧组、高氧+OSI -027组和高氧+雷帕霉素组。
Western blot 检测α-平滑肌肌动蛋白(α-smooth muscle actin , α-SMA )、I 型胶原蛋白(collagen type I , Col I )、增殖细胞核抗原(proliferating cell nuclear antigen , PCNA )、细胞周期蛋白D1(cyclin D1)、RhoA 、Rho 相关含卷曲螺旋蛋白激酶1(Rho -associated coiled -coil -containing protein kinase 1, ROCK1)、蛋白激酶B (protein kinase B , PKB/AKT )、p -AKT 和mTOR 的表达; CCK -8实验检测细胞活力;流式细胞术检测细胞周期。
结果:与对照组相比,PCNA 、cyclin D1、Col I 和α-SMA 表达随高氧处理时间增加而增加(P <0.05)。
与高氧组相比,OSI -027及雷帕霉素干预后,细胞活力下降,细胞周期被抑制在G 1期(P <0.05)。
细胞培养用青霉素-链霉素产品说明书
细胞培养用青霉素-链霉素产品简介:细胞培养用青霉素-链霉素(Penicillin-Streptomycin for Cell Culture)为粉剂,是最常用的细胞培养用抗生素(即通常所谓的双抗)。
在细胞培养液中推荐的青霉素的工作浓度为100U/ml ,链霉素的工作浓度为0.1mg/ml 。
一个包装的细胞培养用青霉素-链霉素可以配制80L 细胞培养液。
保存条件:室温保存。
4ºC 保存可以使用更长时间。
注意事项:开瓶后需防止受潮。
本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
为了您的安全和健康,请穿实验服并戴一次性手套操作。
使用说明:细胞培养用青霉素-链霉素可以参考如下两种方法之一使用:1. 配制细胞培养液时加入细胞培养用青霉素-链霉素,然后再过滤除菌:配制细胞培养液时按照青霉素的工作浓度为100U/ml ,链霉素的工作浓度为0.1mg/ml 进行配制,配制完成后过滤除菌即可使用。
2. 配制青霉素-链霉素溶液(100X)母液,然后再添加到细胞培养液中:按照青霉素的含量为10KU/ml ,链霉素的含量为10mg/ml ,配制青霉素-链霉素溶液(100X)母液。
过滤除菌后即可按照100倍稀释加入到细胞培养液中使用。
配制的母液可以-20ºC 冻存。
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BAG2 amelioratesendoplasmic reticulum stress-induced cell apoptosis in Mycobacterium tuberculosis-infected macrophages through selective autophagy.Autophagy. 2019 Nov 11:1-15.68.Yuan K,Lai C,Wei L,Feng T,Yang Q,Zhang T,Lan T,Yao Y,XiangG,Huang X. The Effect of Vascular Endothelial Growth Factor on Bone Marrow Mesenchymal Stem Cell Engraftment in Rat Fibrotic Liver upon Transplantation. Stem Cells Int. 2019 Dec 4;2019:5310202.Version 2021.11.04碧云天/Beyotime 400-1683301/800-8283301 ST488 细胞培养用青霉素-链霉素 3 / 3。
中性粒细胞胞外诱捕网放大脂多糖诱导的肺泡巨噬细胞炎症反应
中性粒细胞胞外诱捕网放大脂多糖诱导的肺泡巨噬细胞炎症反应赵宁;李勇;邵强;杨云;陈家泉;彭巍;胡世林;钱克俭;刘芬【摘要】Objective To observe the amplification effect of neutrophil extracellular traps (NETs) on inflammatory response ofalveolar macrophages induced by lipopolysaccharide (LPS). Methods Neutrophils were isolated and extracted from peripheral bloodof C57 BL/6 mice by using a mouse peripheral blood neutrophil separation kit. NETs were induced by PMA (100 nmol/L). Observa-tion of NETs by fluorescence microscopy and scanning electron microscopy. NR8383 rat alveolar macrophages were cultured invitro and divided into ⑴controlgroup:adding equal volume PBS;⑵NETs group:adding prepared (1.7×106 cell/ml) NETs;⑶LPSgroup: adding 1 mg/L LPS; ⑷LPS+NETs group:LPS combined with NETs co-stimulation. All groups were stimulated for 12 hours.Enzyme-linked immunosorbent assay (ELISA) was used to detect the contents of interleukin-1β (IL-1β) and tumor necrosis fac-tor-α (TNF-α) in the supernatant of cells. Results Fluorescence microscopy showed that the nucleus of neutrophils in the controlgroup was complete, and there was no extracellular co-localized reticular structure between DNA and NETs. DNA reticular struc-ture could be seen in PMA stimulation group and co-localized with NETs. Scanning electron microscopy also showed that thechromatin of neutrophils was released into extracellular structure, suggesting that the formation of NETs was successfully inducedby PMA.LPS and NETs were used to stimulate alveolar macrophages for 12 hours respectively, the contents of IL-1β and TNF-αin supernatant of alveolar macrophages were significantly higher than those in control group.The contents of IL-1β in supernatantof LPS+NETs group were significantly higher than those of LPS group (P<0.05). There was no significant difference in TNF-α insupernatant between LPS +NETs group and LPS group (P>0.05). Conclusion NETs can amplify LPS-induced inflammation inalveolar macrophages. NETs increased LPS-induced alveolar macrophage inflammatory factor release, suggesting that NETs canamplify LPS-induced alveolar macrophage inflammatory response.%目的观察中性粒细胞胞外诱捕网 (NETs) 对脂多糖 (LPS) 诱导的肺泡巨噬细胞炎症反应的放大作用.方法采用小鼠外周血中性粒细胞分离试剂盒分离提取C57BL/6小鼠外周血中性粒细胞.采用佛波酯 (PMA) (100nmol/L) 诱导中性粒细胞形成NETs, 采用荧光显微镜及扫描电子显微镜观察NETs.体外培养大鼠肺泡巨噬细胞NR8383, 分为⑴对照组:加入等体积PBS;⑵NETs组:加入制备好的 (1.7×106个/ml) NETs;⑶LPS组:加入1mg/L LPS;⑷LPS+NETs组:LPS联合NETs共刺激, 各组均刺激12h.采用酶联免疫吸附试验 (ELISA) 检测细胞上清液中白细胞介素-1β (IL-1β) 、肿瘤坏死因子-α (TNF-α) 含量.结果荧光显微镜下观察示对照组中性粒细胞核形态完整, 无DNA与NETs的胞外共定位网状结构, PMA刺激组可见DNA网状结构, 且与NETs共定位;扫描电子显微镜下也观察到中性粒细胞染色质呈网状结构释放至胞外, 提示PMA诱导NETs形成成功.采用LPS、NETs分别刺激肺泡巨噬细胞12h, 细胞上清液中的IL-1β、TNF-α含量均较对照组显著升高, LPS+NETs组上清液中的IL-1β含量较LPS 组显著升高 (P<0.05), 上清液中的TNF-α较LPS组相比差异无统计学意义 (P>0.05).结论 NETs增加LPS诱导的肺泡巨噬细胞炎症因子释放, 提示NETs可以放大LPS诱导的肺泡巨噬细胞炎症反应.【期刊名称】《江西医药》【年(卷),期】2019(054)001【总页数】4页(P23-26)【关键词】中性粒细胞胞外诱捕网;肺泡巨噬细胞;炎症反应白细胞介素-1β;肿瘤坏死因子-α【作者】赵宁;李勇;邵强;杨云;陈家泉;彭巍;胡世林;钱克俭;刘芬【作者单位】南昌大学第一附属医院重症医学科, 南昌 330006;南昌大学第一附属医院肿瘤科, 南昌 330006;南昌大学第一附属医院重症医学科, 南昌 330006;南昌大学第一附属医院重症医学科, 南昌 330006;南昌大学第一附属医院重症医学科, 南昌 330006;南昌大学第一附属医院重症医学科, 南昌 330006;南昌大学第一附属医院重症医学科, 南昌 330006;南昌大学第一附属医院重症医学科, 南昌 330006;南昌大学第一附属医院重症医学科, 南昌 330006【正文语种】中文【中图分类】R-332中性粒细胞胞外诱捕网(NETs)是中性粒细胞受到炎症刺激后,细胞核内组分如解聚的DNA、组蛋白和颗粒蛋白等释放至细胞外形成的一种网状纤维结构[1]。
罗伯茨绿僵菌线粒体基因组的测序及注释分析
罗伯茨绿僵菌线粒体基因组的测序及注释分析线粒体基因组因为快速进化、严格遵守母系遗传等特点已广泛应用到遗传结构与系统分类的生物学研究,是研究真菌系统进化与遗传关系的有效工具。
作为昆虫病原真菌中已有普遍应用的绿僵菌属真菌(Metarhizium),其线粒体基因组数据尚不完整。
为了进一步完善绿僵菌属线粒体基因组数据,深入昆虫病原真菌遗传与进化的研究,本论文选取罗伯茨绿僵菌(Metarhizium robertsii)ARSEF 2575,采用PDA固体培养基培养、CTAB法提取总DNA,经高通量测序、PCR扩增、Sanger测序成功组装其线粒体基因组并进行注释分析,结合在NCBI上已有的麦角菌科真菌的相关数据,开展比较线粒体基因组学比较,对17种肉座菌目真菌的14个常见的线粒体蛋白的氨基酸序列进行系统发育关系的重建。
结果如下:罗伯茨绿僵菌ARSEF 2575的完整线粒体基因组大小为24945 bp,包含14个常见蛋白编码基因、2个核糖体RNA基因和25个转运RNA基因,蛋白编码基因的种类和排列顺序与已经报道的麦角菌科真菌基本一致。
此外,同多数真菌相似,罗伯茨绿僵菌线粒体基因组的蛋白编码基因、tRNA 基因以及核糖体RNA基因均有明显的A+T偏好性。
通过分析其蛋白编码基因密码子的3位碱基的组成,发现在密码子中,位于第1位点的A、T含量相差较小,而在第2位点的T含量明显比A多出一倍之多,第3位点的A含量是三个位点中最高的,A+T总占比达到83.4%。
在14个蛋白编码基因均以ATG起始并以TAA结束,并没有发现其他起始或终止密码子。
在罗伯茨绿僵菌氨基酸组成中,亮氨酸的使用频率最高,其次为异亮氨酸、苯丙氨酸以及丝氨酸,四者共占线粒体基因氨基酸总量的42.93%。
选取数据库上已发表的肉座菌目真菌以及罗伯茨绿僵菌,基于14个蛋白质编码基因的氨基酸序列,采用最大似然法,建立系统发育树。
所得拓扑结构与目前已知的肉座菌目的分类基本一致,显示Metarhizium robertsii与Metarhizium anisopliae亲缘关系最近。
肝病专业英语词汇
3α-羟类固醇脱氢酶(Y' 蛋白) γ -谷氨酰转移酶 γ-氨基丁酸 甲胎蛋白 人兽共患病 异种肝移植 脂肪性纤维瘤,黄色瘤 黄嘌呤氧化酶 黄斑瘤 全球移植中心名录 窗口期 肝豆状核变性 肥达反应 外斐反应 韦克斯勒成人智力测验 呕吐 视觉诱发电位 病毒学应答 病毒复制 病毒性肝炎 静脉-静脉转流 VOD 肝小静脉闭塞病 静脉-动脉转流 血管活性肽 静脉曲张 胆管消失综合征 疫苗 熊去氧胆酸 尿胆素原 尿胆素 二磷酸尿苷异构酶 尿素生成 鸟氨酸循环,尿素循环 上消化道出血 粗纤维调节素 肝未分化肉瘤 充盈不足学说 非结合高胆红素血症 游离胆红素,非结合胆红素 超声(波)检查法
短潜伏期肝炎 移动性浊音 腹水白蛋白浓度梯度 血清肝炎 血清诊断 血清胆红素 血清白蛋白 血清学应答 乙肝血清学检查 血清转换 败血症相关胆汁淤积 正链,有义链 镇静剂 次级胆酸 海蓝组织细胞增多症 硬化疗法 硬化性胆管炎 日本血吸虫病 湄公血吸虫 曼氏血吸虫 日本血吸虫 间插血吸虫 埃及血吸虫 血吸虫 瘢痕形成期 粗面内质网 滚环机制(环状DNA复制的机理) RNA干扰 核酶 核糖体 胆固醇逆向转运
伤寒 Ⅳ型胶原 Ⅲ型前胶原 甲肝和乙肝疫苗 抑癌基因 肿瘤坏死因子 肝结核 滋养体 甘油三酯 颠换效应 经颈静脉肝内门腔分流 转换 输血传染的病毒 输血性肝炎 转化生长因子
transcatheter arterial chemoembolization 肝动脉化疗栓塞 trans-activation toxic hepatitis total cholesterin total bilirubin tomor necrosis factor tocopherol TNF-related apoptosis inducing ligand tissue inhibitor of metalloproteinase thymosin Thymopolypeptides for Injection thromboxane the core promoter element tentative diagnosis tension of muscle tenderness Telbivudine taurocholic acid taurochenodeoxycholate acid systemic inflammatory response syndrome syncytial giant-cell hepatitis sustained virus response sustained response 反式激活 中毒性肝炎 总胆固醇 总胆红素 肿瘤坏死因子 生育酚,维生素E 肿瘤坏死因子相关凋亡诱导配体 基质金属蛋白酶组织抑制物 胸腺肽 胸腺肽 血栓素 核心启动子元件,启动子核心元件 暂时的(假定的)诊断,试验性诊断 肌张力 压痛 LdT 牛(磺)胆酸 牛磺鹅(去氧)胆酸盐,牛磺鹅(脱氧)胆酸盐 全身炎症反应综合症 融合巨细胞性肝炎 持续病毒应答 持久应答
crispr原理解析经典实用
Natronobacterium gregoryi Argonaute (NgAgo)是一种DNA导向的可用于人类细胞基因编辑 的核酸内切酶,与Cas9不同,NgAgo–gDNA系统不需 要PAM,初步鉴定表明,该系统对导向-靶向 (guide–target)错配耐受低,且对编辑富含G+C的 基因组更加有效。据介绍,Cas9只存在于原核生物中, 而Argonautes几乎存在于所有的有机体中。要想与 Cas9正确绑定,导向RNA必须有3′RNA-RNA杂化结构, 而与Argonaute绑定不需要导向分子有任何特定的二 级结构。另一方面,Cas9只能切割PAM上游的序列, 而Argonaute不需要靶标有特定的序列。 NgAgo有望 成为编辑哺乳动物基因组的精准有效的工具。
crispr原理解析
• crRNA:当细菌抵御噬菌体等外源DNA ( protospacers )入侵时,在前导区的调控下, CRISPR被转录为长的RNA前体(pre-crRNA),然后加 工成一系列短的含有保守重复序列和间隔区的成 熟crRNA,pre-crRNA转录的同时,与其重复序列 互补的反式激活crRNA (Trans-activating crRNA, tracrRNA)也转录出来。
crispr原理解析
• 敲除AIP1基因的CRISPR/Cas9慢病毒系统,成 功获得AIP1敲除的人(胚肾细胞株)293T稳定 细胞株(广州医科大学)
• 在一个黑素瘤模型中,筛选出了涉及药物 维罗非尼(Vemurafenib)耐药性的基因 (麻省理工学院 张峰)
crispr原理解析
CRISPR/Cas9的靶向特异性是由两部分决定的, 一部分是RNA嵌合体和靶DNA之间的碱基配对,另一 部分是Cas9蛋白和一个短DNA基序(DNA motif)的结 合,这个短DNA基序通常在靶DNA的3‘末端发现,被 称为前间区序列邻近基序(protospacer adjacent motif, PAM)
实验六农杆菌介导转化拟南芥
Hyg基因或Lac Z基因等。
双元载体系统旳转化原理是Ti质粒上 旳Vir基因能够反式激活T-DNA 旳转移。
外外源源基基因因 切
3 转化措施
• 根据受体材料不同分为浸花法、原生质 体共培养法、叶盘法和创伤植物感染法。
• 其中 浸花法、叶盘法在许多植物上得 到广泛应用。
试验流程
1. 挑取活化旳含目旳基因质粒旳单克隆阳性农杆菌GV3101菌 株至5ml 新鲜YEB液体培养基(50g/ml Kan,125g/ml Rif)中,28℃摇培24h。
科植物,二年生草本,高7~40厘米,花期3~5 月。广泛用于植物遗传学、发育生物学和分子生 物学旳研究,已成为一种经典旳模式植物,其原 因主要基于这种植物具有下列特点:
• (1)形态个体小,生活力强,种子量大;
• (2)生长周期快,从播种到收获种子一般只需 6--8周;
• (3)基因组小(2n=10),是目前已知植 物基因组中最小旳,但是具有完毕“复 杂”旳植物生长发育旳全部基因;
试验六、农杆菌介导转化拟南芥
试验目旳:学习真核生物旳转基因技术及农杆菌介 导旳转化原理。
•试验要求:掌握农杆菌介导转化拟南芥 旳试验措施, 了解拟南芥旳生理特点及在基因工程试验中应用。
• 技术应用:一种以便,高效旳植物转基因措 施,多用于转化双子叶植物,经过改善后也 逐渐用于单子叶植物旳转化。
• 试验材料: 农杆菌GV3101重组菌株,重组双元 体现载体pCAMBIA1300 ,拟南芥
Vir区
2.3 双元体现载体系统源自•双元体现载体系统主要涉及两个部分:
•
一部分为卸甲Ti质粒,此类Ti质粒因为缺失了T
-DNA 区域,完全丧失了致瘤作用,主要是提供Vir
构建下转换荧光-适配体的免疫层析试纸条用于快速检测黄曲霉毒素B_(1)
构建下转换荧光-适配体的免疫层析试纸条用于快速检测黄曲霉毒素B1王邹璐琪1,李立煌1,李丹阳1,艾超超1,任磊1,*,孙本强2,*(1.厦门大学材料学院,福建厦门361005;2.厦门医学院附属口腔医院,福建厦门361005)摘 要:构建下转换荧光-适配体免疫层析试纸条用于食品中黄曲霉毒素B1(aflatoxin B1,AFB1)的快速高效检测。
体系中AFB1存在会减弱下转换荧光-适配体纳米颗粒层析至T线时与AFB1半抗原的结合能力,从而导致下转换荧光信号衰减,进而实现对AFB1的高效检测。
该方法在AFB1质量浓度1~40 ng/mL范围内与荧光信号呈良好的线性关系,线性相关系数为0.994,检测限为0.287 ng/mL。
该方法利用稀土掺杂荧光纳米颗粒的长寿命发光及近红外荧光特性,有效降低了生物背景荧光干扰并提高了检测体系的特异性。
该方法在AFB1的快速高灵敏检测中具有良好的应用前景。
关键词:稀土掺杂荧光纳米颗粒;荧光免疫层析;黄曲霉毒素B1;快速检测Construction of Down-conversion Fluorescence-Aptamer Immunochromatographic Strip for Rapid Detection of Aflatoxin B1 WANG Zouluqi1, LI Lihuang1, LI Danyang1, AI Chaochao1, REN Lei1,*, SUN Benqiang2,*(1. College of Materials, Xiamen University, Xiamen361005, China;2. Stomatological Hospital of Xiamen Medical College, Xiamen361005, China)Abstract: In this study, a down-conversion fluorescence-aptamer immunochromatographic strip was constructed for the rapid and efficient detection of aflatoxin B1 (AFB1) in foods. The presence of AFB1 in the system will weaken the binding ability of down-conversion-aptamer fluorescent nanoparticles to the hapten AFB1 when down-conversion-aptamer fluorescent nanoparticles reach the T-line, thus leading to the attenuation of down-conversion fluorescence signal and consequently highly efficient detection of AFB1. In the range of 1–40 ng/mL, the concentration of AFB1 had a good linear relationship with the fluorescence signal, showing a correlation coefficient of 0.994, and the detection limit for AFB1 was0.287 ng/mL. By taking advantage of the long-lived luminescence and the near infrared fluorescence characteristics of rareearth doped fluorescent nanoparticles, this method effectively reduced the interference of biological background fluorescence and improved the specificity of the detection system, making it a promising candidate for application in the rapid and sensitive detection of AFB1.Keywords: rare earth doped fluorescent nanoparticles; fluorescence immunochromatographic assay; aflatoxin B1; rapid detection DOI:10.7506/spkx1002-6630-20191030-337中图分类号:TS201.2 文献标志码:A 文章编号:1002-6630(2021)12-0295-07引文格式:王邹璐琪, 李立煌, 李丹阳, 等. 构建下转换荧光-适配体的免疫层析试纸条用于快速检测黄曲霉毒素B1[J]. 食品科学, 2021, 42(12): 295-301. DOI:10.7506/spkx1002-6630-20191030-337. WANG Zouluqi, LI Lihuang, LI Danyang, et al. Construction of down-conversion fluorescence-aptamer immunochromatographic strip for rapid detection of aflatoxin B1[J]. Food Science, 2021, 42(12): 295-301. (in Chinese with English abstract) DOI:10.7506/spkx1002-6630-20191030-337. 收稿日期:2019-10-30基金项目:福建省自然科学基金项目(2017Y0078);国家自然科学基金面上项目(31870994)第一作者简介:王邹璐琪(1996—)(ORCID: 0000-0002-7715-1267),女,硕士研究生,研究方向为生物医学材料。
基因芯片技术原理
基因芯片(Gene chip)
Southern & Northern Blot Dot Blot
Macroarray
Microarray
基因芯片的研发历史
八十年代末期俄美科学家提出“杂交法测序”
1992年世界第一块原位合成基因芯片在美国 Affymetrix诞生(1994第一张商业化芯片)
1995年世界第一块微矩阵基因芯片在Stanford大 学实验室诞生
技术背景
随着传统技术的不断改进,基因信 息分析规模不断扩大。 • -人类基因组计划的需求(HGP) • -后基因组时代的需求(功能基因 组学方向)
生物芯片 (Biochips)
将大量生物识别分子按预先设置的排列固定于一种载 体(如硅片、玻片及高聚物载体等)表面,利用生物分子 的特意性亲和反应,如核酸杂交反应,抗原抗体反应等来 分子各种生物分子存在的量的一种技术。
image
Commercial chip
• Human • Rat • Mouse • Drosophila • E. coli • Yeast • Zebrafish
基因芯片的应用
– 特定基因检测 – 突变检测 – 多态性分析 – 基因表达谱
•生物信息学的工具 •基因相关性研究 •基因功能 •药物设计和开发 •潜在反义试剂开发 •个体化医疗 •身份识别 •基因诊断 •其他与生物有关的领域
Removable Tip Orifice
Controller
直接点样法 (stanford, brown)
Spotter head
基因芯片技术流程
基因芯片的设计制备 杂交 检测
数据分析
制备方法及点样仪器
靶基因的标记制备:标记方法 杂交:杂交液、杂交温度、洗涤条件
液相色谱串联三重四极杆质谱的使用、常见故障处理与维护保养
第 30 卷第 2 期分析测试技术与仪器Volume 30 Number 2 2024年3月ANALYSIS AND TESTING TECHNOLOGY AND INSTRUMENTS Mar. 2024大型仪器维护与维修(130 ~ 137)液相色谱串联三重四极杆质谱的使用、常见故障处理与维护保养许晓辉,李 坚,王小乔,吴福祥,潘秀丽,李 赟,李 运,王月玲,邵长春,张生萍(兰州市食品药品检验检测研究院/国家市场监管重点实验室(食品中农药兽药残留监控),甘肃兰州 730050)摘要:液相色谱串联三重四极杆质谱(liquid chromatography tandem triple quadrupole mass spectrometry, LC-MS/MS)广泛应用于环境分析、食品安全、药物代谢研究、生物医学等领域,主要用于复杂样品基质中痕量目标物检测,既适用于小分子有机化合物定性,又可用于痕量化合物定量. 其主要检测模式是多反应监测模式(multiple reaction monitoring, MRM),以定性离子对的丰度比来进行定性,以定量离子对的峰响应进行定量,在MRM检测模式下具有出色的检测灵敏度和定量功能. LC-MS/MS的合理规范使用与维护保养,是保证检测数据准确有效、仪器可持续正常运行的基本要求,也是降低仪器维修成本的重要举措. 阐述了安捷伦LC-MS/MS的工作原理、开机与关机、调谐,探讨了常见的故障及应对措施、日常维护保养的内容,旨在为合理规范使用仪器提供参考.关键词:液相色谱串联三重四极杆质谱仪;三重四极杆;使用;故障处理;维护保养中图分类号:O657. 63 文献标志码:B 文章编号:1006-3757(2024)02-0130-08DOI:10.16495/j.1006-3757.2024.02.009Use, Troubleshooting and Maintenance of Liquid Chromatography Tandem Triple Quadrupole Mass SpectrometryXU Xiaohui, LI Jian, WANG Xiaoqiao, WU Fuxiang, PAN Xiuli, LI Yun, LI Yun, WANG Yueling, SHAOChangchun, ZHANG Shengping(Lanzhou Institute for Food and Drug Control/Key Laboratory of Pesticides and Veterinary Drugs Monitoringfor State Market Regulation, Lanzhou 730050, China)Abstract:The liquid chromatography tandem triple quadrupole mass spectrometry (LC-MS/MS) is widely used in environmental analysis, food safety, drug metabolism research, biomedicine and other fields. It is mainly used for the detection of trace targets in complex sample matrix, which is suitable for the qualitative of small molecular organic compounds and the quantitative of trace compounds. The main detection mode is multiple reaction monitoring (MRM). It has excellent detection sensitivity and quantitative function in MRM mode by abundance ratio of qualitative ion pair to qualitative and peak response of quantitative ion pair to quantitative. Reasonable standard use and maintenance of the LC-MS/MS is the basic requirement to ensure accurate and effective test data and sustainable normal operation of the收稿日期:2023−12−11; 修订日期:2024−03−07.基金项目:2022年度兰州市第二批科技计划项目(2022-2-45),兰州市科技发展指导性计划项目(2023-ZD-231),甘肃省药品监督管理局科学技术类项目(2022GSMPA0068)作者简介:许晓辉(1986−),男,硕士,高级工程师,主要从事食品药品检验检测,E-mail:******************通信作者:王小乔(1973−),女,本科,高级工程师,主要从事食品药品检验检测,E-mail:****************;张生萍(1971−),女,本科,高级工程师,主要从事食品药品检验检测,E-mail:*****************.instrument, and is also an important measure to reduce the maintenance cost of the instrument. The operating principle, power-on, power-off, tune about the LC-MS/MS of Agilent were described, and the common faults and countermeasures, as well as the contents of daily maintenance were discussed in order to provide a reference for the rational and standardized use of the instrument.Key words:LC-MS/MS;triple quadrupole;use;troubleshooting;maintenance当前,液相色谱串联三重四极杆质谱(liquid chromatography tandem triple quadrupole mass spectrometry,LC-MS/MS)已广泛应用于定性定量检测保健品及中成药违法添加药物[1]、中成药真伪鉴别[2]、农作物和禽畜肉等食品中农兽药残留[3-6]、药物代谢及药物动力学研究[7]、临床药理学研究[8]、天然药物开发研究[7]、新生儿筛选[9]、蛋白与肽类的鉴定[8]、毒物分析[10]、环保[11]、食品[12-13]、化妆品[14]、自来水[15]、海水[16]、卫生防疫[17]等行业,其具有高分离能力、高选择性、高灵敏度、检测限低、分析速度快及能够提供相对分子质量与结构信息等优点. 研究表明,60%的仪器故障都可归因于缺乏预防性维护,如果实验室采取了预防性维护措施,可将机械系统的故障率降低,因此规范使用与持续性维护保养是延长仪器使用寿命和保证数据结果真实性、准确性的关键工作. 在实际工作中,仪器操作不当和维护保养不到位,都会导致仪器发生故障,一旦损坏,厂家上门维修时,不仅增加仪器使用成本,而且影响工作进度. 因此,正确规范使用及维护保养仪器尤为重要,将会延长仪器正常运行时间、最大限度提高效率并延长仪器寿命. 本文从安捷伦LC-MS/MS的基本原理、开机和关机、调谐、常见故障分析和基本维护保养方面进行论述,以期为LC-MS/MS规范使用与维护保养提供参考.1 基本原理LC-MS/MS由液相、质谱、真空和载气部分组成. 液相部分分离被测物,质谱部分检测碎片离子,样品经液相部分分离后进入质谱部分被离子化,质谱的质量分析器会根据离子碎片质量数大小按质荷比将其分开,得到一级质谱图和二级质谱图. LC-MS/MS构造主要包括离子源、毛细管、离子光学组件、四极杆1(Quadrupole 1,Q1)和四极杆3(Quadrupole 3,Q3)、碰撞池(Quadrupole 2,Q2)、检测器和真空系统. 一般粗真空在199.983~333.305 Pa (1.5~2.5 Torr,1 Torr=133.322 Pa)之间,碰撞气关闭时高真空在 3.999×10−4~7.999×10−4Pa(3×10−6~ 6×10−6 Torr)之间,碰撞气打开时高真空在2.533×10−3~3.066×10−3 Pa(1.9×10−5~2.3×10−5 Torr)之间.离子源是LC-MS/MS的“心脏”,电喷雾离子源(electron spray ionization,ESI)是目前应用极为广泛的电离方式,其工作原理是:雾化器的喷雾针被带高电压的半圆柱体形电极环绕,带有被测物质离子的流动相在雾化针尖端发生雾化,由于半圆形电极和毛细管间的电压不同,产生的电场使液滴表面富集带同种电荷的离子,而内部带相反电荷的离子聚集,形成带电液滴的细喷雾,液滴在电场的作用下,飞向毛细管. 加热的氮气干燥气体反向流动,带走液滴中的中性溶剂分子,从而收缩液滴,直到排斥的静电力超过液滴表面张力,引起库仑爆炸. 这个过程不断重复,直到待分析物离子最终变成气态进入毛细管. 带有安捷伦喷射流技术的ESI源与普通ESI源的电离原理相同,提供和普通ESI源相同的离子特征,但在喷射流技术中增加了加热的鞘流气,并可以在喷嘴处施加喷嘴电压,更高的温度可以提高流动相的蒸发率,有利于气溶胶的形成和液滴的蒸发,减少喷雾中的大液滴. 由于使用了热梯度聚焦技术,同轴的鞘流气有助于减少离子扩散,将更多的离子引入到质谱(mass spectrometry,MS)中,减少中性溶剂束进入MS,提高了灵敏度,降低了噪音.三重四极杆质谱的四极杆分析器由4根棒状电极组成,其互相平行且距离相等,构成四极电场,四极电场对角位置的2根电极被连接在一起,其中一对同时施加直流电压和射频电压,而另一对施加极性相反、大小相同的直流电压和振幅/频率相同、相位相反的射频电压. 射频电压和直流电压相互叠加,且两对电极不停地快速切换极性,在四极杆之间形成动态电场,使离子呈螺旋轨道运动. Q1和Q3由直流电(direct current,DC)和射频电(radio frequency,RF)控制,Q2仅由允许所有离子通过的RF控制. 对于给定的DC和RF组合,由于振幅是第 2 期许晓辉,等:液相色谱串联三重四极杆质谱的使用、常见故障处理与维护保养131规定的,只有特定质荷比的离子才能通过四极杆到达检测器,其他离子无法通过四极杆. 基于工作原理,三重四极杆质谱有4种常见数据采集扫描模式,分别是全扫描(MS2 Scan)、选择性离子扫描(selected ion monitor,SIM)、子离子扫描(pro ion scan)、多反应监测模式扫描(multiple reaction monitoring,MRM).2 开机和关机2.1 开机如果使用液氮罐,打开液氮罐自增压阀门,调节液氮罐分压表的输出压强为约0.7 MPa(~100 psi,1 MPa=145.037 psi),并检查前级泵的镇气阀状态,使其处于关闭. 如果使用氮气发生器,打开气路开关,再打开氮气发生器电源,工作半小时后,调节输入LC-MS/MS氮气压力表数值约为0.7 MPa(~100 psi),并确认前级泵镇气阀处于关闭状态. 使用氮气发生器,请务必依照相应厂家建议,定期维护,更换过滤芯等,防止氮气污染质谱,影响仪器性能. 打开高纯氮气主阀门,调节高纯氮气钢瓶次级减压表输出压强至0.15 MPa(~22 psi),最大不要超过0.2 MPa(~29 psi). 依次打开计算机、液相色谱仪各个模块电源、质谱仪电源开关,机械泵开始工作,仪器开始自检,等待大约5 min,仪器自检完成,这时进入MassHunter工作站监视真空泵的真空值,当真空值达到规定值且Turbo1和Turbo2涡轮泵的转速达到100%之后,质谱状态信号灯变为绿色,仪器才能使用. 一般来说停机较长时间后开机时,需要抽真空至少6 h后才可分析样品.2.2 关机在MassHunter采集软件内右键点击三重四极杆MS的图标,下拉图中选择“Vent”,确认要放空,选择“Yes”,可以在三重四极杆Method的Diagnosis界面观察涡轮泵转速的下降情况. 分子涡轮泵转速和功率基本为0后,等待30 min,关闭MassHunter软件. 然后关闭质谱、液相色谱仪各模块及电脑的电源. 需注意的是:如果长时间关机请拔掉质谱主电源线,关闭氮气室气体通路开关. 如果使用液氮罐,关闭液氮罐增压阀门. 如果使用氮气发生器,关闭氮气发生器电源开关. 建议不要关闭碰撞气高纯氮减压阀,在仪器关机后,并不会消耗高纯氮,但可以使整个管路保持正压,有效保护高纯氮的捕集阱不被环境空气污染.3 调谐调谐包括3种方式:自动调谐(autotune)、检验调谐(checktune)和手动调谐(manual tune). 自动调谐是在整个质量范围内自动调节质谱各种参数,使其获得最佳性能参数. 检验调谐是以当前调谐文件参数采集数据,确认和评价质谱参数是否发生偏移,不改变当前质谱参数. 手动调谐是手动改变质谱性能各个参数. 自动调谐步骤:右键点击采集软件中三重四极杆图标,选择“On”. 在Context下拉框中选择“Tune”,选择切换到Tune界面,等待三重四极杆图标变绿,达到Ready状态后,如果是Jet Steam源,请继续等待15~30 min,然后再进行调谐,如果是ESI源,无需等待可以直接进行调谐. 点击“Autotune”页面,上部是极性选择和自动调谐选项,可以对正、负极性分别做调谐或者选择正负极性同时调谐. 一般情况下,不必选“Both”,只要调谐需要的极性即可. “Start from factory default”选项大部分情况不必选择,不选择此项时,调谐参数优化从上次调谐结果开始,选择此项则自动调谐会从工厂默认参数开始. 点击“Autotune”按钮,调谐液会自动进入质谱,自动开始调谐. 自动调谐结束后,底部显示调谐完成,并自动弹出调谐报告,调谐文件会自动保存为“atunes.Tune.xml”. 调谐结束后,调谐液会自动关闭. 检验调谐是调用当前调谐文件参数来采集数据,确认和评价仪器是否有偏移,通过检验调谐结果来判别质谱是否存在问题,并不改变质谱各个参数. 如果检验调谐不通过,还可以对质谱进行自动调谐. 需注意的是,“Jet Steam”在“ON”状态下不要直接打开离子源,将质谱检测器设置为“Standby”状态,等待温度下降至125 ℃后,才能打开离子源,避免烧坏离子源加热组件. 三重四极杆质谱一般较稳定,不需要经常进行调谐,或者在“Checktune”不能通过时进行调谐,自动调谐周期一般2~3个月即可.调谐是否通过,主要通过调谐报告中一些参数来判断,调谐报告都会详细记录调谐质谱的基本情况,包括调谐时真空读数、调谐时离子源参数、质量准确度差值、半峰宽差值、调谐离子响应等(如图1所示). 其中,质量准确度差值和半峰宽差值直接决定了调谐及检验调谐是否通过,质量准确度差值132分析测试技术与仪器第 30 卷(Delta)指实测质荷比(mass to charge ratio,m/z)和目标m/z的差值,也就是质量准确度,一般小于0.2即认为正常. 半峰宽差值(delta)指实测半峰宽(full width at half maxima)和目标半峰宽的差值,也就是分辨率,一般“unit”与0.7的差值在0.14之内、“wide”与1.2的差值在0.6之内、“widest”与2.5的差值在1.25之内. 有时调谐报告显示黄色的“Adjust”,如果目标分析物m/z范围在该质量数以下,可以直接用,不受影响. 如果目标分析物m/z 范围在该质量数范围内,则必须进行调谐,才可测样. 有时虽然调谐结果全通过,显示绿色,但此时也要关注调谐离子响应,如果本次调谐与本机之前调谐离子响应相比,下降不多则认为仪器正常,如下降很多则认为仪器不正常,此时造成仪器响应下降的原因有很多,主要有离子源、喷雾针、毛细管和调谐液,建议维护离子源和喷雾针,清洗毛细管或更换毛细管,更换新的调谐液.4 常见故障分析4.1 不出峰不出峰应查看喷雾、真空和电流. (1)由于样品是由离子源喷雾进入质谱,所以第一步应查看喷雾.打开手电筒,将光通过另一侧视窗照进离子源内部,在离子源的视窗处,通过放大镜来检查喷雾针状态,正常状态下喷雾应是细密的伞状连续喷雾,非正常状态下喷雾不连续,甚至是没有喷雾. (2)观察采集软件质谱参数状态栏,查看真空.粗真空应在266.644 Pa(2 Torr)左右,高真空在0.001 3 Pa(10−5 Torr)左右. 如果粗真空参数较好,可能是毛细管堵塞,如果粗真空参数无法达到,可能是由于机械泵故障. 如果高真空参数较好,可能是由于碰撞气体不足导致. (3)查电流和毛细管电流. 这两个反映的是离子化效率,仪器稳定时,电流值应是稳定的,如果发现电流值降低,则可能是离子源受到污染,导调谐时真空读数调谐时离子源参数调谐离子响应质量准确度差值半峰宽差值m/z m/zm/z m/zm/z m/zm/z m/z图1 调谐报告Fig. 1 Tuning report第 2 期许晓辉,等:液相色谱串联三重四极杆质谱的使用、常见故障处理与维护保养133致离子化效率降低.4.2 漏液漏液分为管路漏液和进样器漏液. 当柱压出现明显下降,仪器一定出现了漏液. 管路漏液时,流出的液体会触发仪器报错,但进样器漏液仪器往往不会报错,因而不易察觉. 此外,进样器漏液往往会导致样品无法部分或完全进入液相系统从而造成不出峰. 为进一步验证进样器是否存在漏液,可以通过切换进样器的六通阀来判断. 当进样器处于主路时,来自泵的液体从六通阀1号位进入,2号位流出,然后经过计量泵、定量环、针和针座进入到六通阀5号位,从6号位流出进入柱温箱. 当进样器处于旁路时,来自泵的液体从六通阀1号位进入,6号位直接流出到柱温箱. 因此,进样器在主路时经过部件较多,所以进样器处于主路时的压强理论上要比在旁路高. 切换进样阀,如果主路压强比旁路的高,则进样系统不会有太大漏液,如果主路压强与旁路持平或更低,则进样器可能存在漏液.4.3 氮气不足引发仪器报错由于氮气发生器压强不足导致质谱氮气供应不足,引起仪器报错. 解决方法:关闭工作站,关闭液相部分所有模块电源,重启质谱(直接关闭电源开关停5 s后再开启电源),待质谱自检完毕后,开启液相,待液相自检完毕后,重新打开工作站. 或者把连接质谱和液相的数据线“romote”拔掉,重启后再插上.4.4 针座堵塞判断针座是否堵塞,可以通过查看自动进样器主路和旁路状态压强值来判断. 记录主路状态压强值后,切换到旁路状态,待稳定后再记录其压强值,如果两者压强值相差0.3~0.5 MPa(3~5 bar,1 bar=0.1 MPa)左右,则进样针和针座可能没有堵塞,如果主路状态比旁路状态压强大较多,则进样针和针座发生了堵塞. 针座堵塞时,常用解决方法是反冲.4.5 质谱响应低质谱响应低时,首先检查被测溶液化学性质,确保样品完全溶解,样品溶液澄清透明,确保样品新鲜且得到正确储存. 其次检查液相压强与质谱状态参数,确保液相管路没有受到污染且质谱运行良好,确保仪器已正确调谐并检查调谐离子响应强度.同时检查离子源、毛细管是否受污染,如受污染,则要清洗或更换毛细管,拆开离子源,对离子源通路进行清洗. 最后检查雾化器中针头位置以及喷雾针是否受样品污染在针头产生絮状物,并在流动相流速合理的条件下查看喷雾状态,检查雾化器端部是否发生损坏.4.6 背景噪音高排除流动相本身噪音高的原因,背景噪音高主要有以下原因:离子源受到污染,质量检测器受到污染,喷雾器污染或者损害或者设置有误,质量过滤器值设置不当,溶剂及流动相选择不当. 因此,若背景噪音高,着重从以上方面排查.4.7 重复性差重复性差直观表现在两个方面,一是色谱峰漂移,二是定量离子对峰面积不稳定. 重复性主要跟流动相流速、压强、调谐、碰撞气压强、气体流速、毛细管、离子源、喷雾等相关. 因此,重复性差主要从以下方面排查:确保干燥气流流速和温度相对于溶剂流速无误,确保溶剂已经彻底脱气,确保液相色谱背压稳定,确保相对于液相色谱流速设置的雾化气体压强足够高,确保雾化器喷雾正常,确保离子源及毛细管没有受到污染.4.8 碎裂结果不理想母离子碎裂结果不理想,主要原因是碎裂电压设置过高. 最佳碎裂电压主要跟被分析化合物的质量数有关,一般碎裂电压最佳值处在化合物质量数三分之一左右. 因此,离子碎裂结果不理想时,需重新优化碎裂电压.4.9 质量精度不佳质量精度是碎裂离子实测质量数与理论质量数的误差. 出现质量精度不佳,则需要重新校准质量轴,同时确保用于调谐的离子涵盖样品离子的质量范围,并且显示出强而稳定的信号,确保干燥气流速和温度相对于溶剂流速无误,确保溶剂已经彻底脱气. 如果以水作为流动相组成部分,确保使用去离子水(> 18.2 MΩ·cm).5 维护与保养LC-MS/MS的维护保养主要包括两大模块,即色谱部分和质谱部分. 色谱部分着重注意样品净化完全彻底、管路冲洗、流动相脱气、流动相达到色谱级要求、水相防微生物、定期更换泵头密封垫、使用安捷伦专用进样瓶等,平时记录液相柱压数值,134分析测试技术与仪器第 30 卷关注柱压波动,及时更换水相,在有效期内使用水相,隔一定时间对水相流动相瓶进行微生物清洗.微生物清洗通过在水相流动相瓶加入水并置于95 ℃水浴锅中煮沸,再以色谱纯甲醇或者乙腈清洗水相流动相瓶,最后用超纯水冲洗干净. 每测试一定数量样品之后,使用50%异丙醇、水、有机相与水等依次对管路进行彻底冲洗. 注意样品进样浓度,进样浓度不宜过大,否则会对离子源造成污染. 同时,建议1~2 µL进样体积,当进样体积过大时,尤其是基质复杂样品,除杂不彻底,会造成进样针堵塞.质谱部分着重关注喷雾针、离子源、真空泵状态及泵油状况、捕集阱、氮气压强、废液等. 使用氮气发生器要保证废液瓶口水平面低于排水口水平面,并按时倾倒废液,按时维护保养氮气发生器. 平时注意记录质谱参数状态,如粗真空、高真空、股电流、毛细管电流,如果发现质谱参数和规定数值有偏差,要及时进行排查,同时要按照规定,定期做好维护保养,具体维护保养的内容如表1所列.表 1 维护保养的内容Table 1 Content of maintenance维护频次事项描述涉及备件或耗材每日必做检查及清洗离子源离子源每天测完样品后都需要使用50%异丙醇/水的混合液清洗,擦拭锥孔时最好180°擦拭,且不要有液体滴落无纺布、棉签检查真空读数打开采集软件,观察仪器采集软件右上方状态参数栏的真空读数N/A每周必做清洗电喷雾雾化室每周清洗离子源可以有效去除前期试验带来的污染喷雾针、绝缘套件、无纺布、棉签超声清洗雾化器取下超声清洗雾化器,并用枪头保护好针尖,分别用异丙醇、甲醇、水等溶剂超声10~15 min喷雾针检查泵每周检查机械泵油位置,保证油面在min和max线中间,查看泵油颜色是否透明,检查机械泵泵滤网是否堵塞泵油、过滤器每月必做检查质谱废液桶当废液桶内液体比较多时,及时把多余的溶液倒掉N/A检查氮气压强是否正常普通氮气的压强控制在0.70 MPa左右(~100 psi),高纯氮气控制在0.15 MPa左右(~22 psi)N/A每半年必做清洗毛细管需注意检查毛细管时需要放真空,导电毛细管应使用超声清洗,具体操作:1 g Alconox清洁粉末放入100 mL量筒,用超纯水充分溶解,将毛细管两头用1 mL移液枪枪头套住,竖直放入充满Alconox溶液的量筒中,确保液面没过毛细管,超声清洗10 min后用超纯水冲洗干净毛细管、清洁粉末更换喷雾针喷雾针露出喷雾器的顶端76.2 µm喷雾针更换泵油半年更换一次泵油及油气过滤芯可以大幅提高真空泵的使用寿命,其中机械泵用AVF45 platinum 0.946 4 L的泵油泵油、油气过滤器每年必做检查及更换气体净化器检查空气过滤网是否堵塞. 每年更换一次氮气过滤器可以有效去除普通氮气的杂质,每年更换氮气捕集阱空气过滤器、氮气过滤器、氮气捕集阱清洗光学组件每年清洗光学组件可以保证仪器性能N/A检查光电/电子倍增器电压从调谐报告里找到电子倍增管(EMV)或光电倍增管(PMT)/微通道板光电倍增管(MCP)电压,离最高限值很近,则需要准备新检测器光电倍增器、电子倍增器注:N/A表示不适用6 结语液相色谱串联三重四极杆质谱仪属于大型贵重精密仪器,应健全仪器管理制度,由专人负责管理,固定仪器操作人员,未经过培训人员不得随意操作仪器. 在实际工作中,要保证仪器处于可持续良好运行状态,延长仪器使用寿命,降低仪器故障,从而降低仪器运行成本. 除了要配备不间断电源(uninterruptible power system,UPS),保证仪器电压稳定持续,还要熟悉掌握仪器规范使用规程,按操第 2 期许晓辉,等:液相色谱串联三重四极杆质谱的使用、常见故障处理与维护保养135作规程使用仪器,定期做好仪器维护保养. 同时,更重要更容易被忽视的是要保证样品除杂干净彻底及澄清,样品溶剂应和流动相比例一致,并在上机进样之前,采取一定的措施方法验证样品是否除杂完全,如动物源性食品中兽药残留提取净化样品,放在冷冻冰箱过夜,如有脂肪没有除去干净,便会在进样瓶瓶底析出. 要掌握样品基质特点及目标检测物浓度,进样体积不宜过大,防止堵塞进样针、色谱柱及污染喷雾针与毛细管. 其他高难度维护维修则需联系厂家工程师上门进行. 总之,使用液相色谱串联三重四极杆质谱仪这种大型精密仪器,一定要进行岗前培训,培训合格之后,持证上岗,按照规范严格操作,定期进行维护保养.参考文献:席彰, 周亚兰, 康靖, 等. 高效液相色谱-三重四级杆质谱快速筛查及定量检测改善睡眠类保健品和中成药中的22个精神类化合物[J ]. 药物分析杂志,2022,8(2):320-328. [XI Zhang, ZHOU Yalan,KANG Jing, et al. High performance liquid chromato-graphy-triple quadrupole mass spectrometry rapid screening and quantitative detection of 22 psychotrop-ic compounds in health care products and Chinese pat-ent medicines for improving sleep [J ]. Chinese Journ-al of Pharmaceutical Analysis ,2022,8 (2):320-328.][ 1 ]李婷, 程显隆, 王郡瑶, 等. 超高效液相色谱-三重四极杆质谱法用于含阿胶中成药中胶类皮源检测研究[J ]. 中国药学杂志,2023,58(3):260-265. [LI Ting, CHENG Xianlong, WANG Junyao, et al. Identi-fication of gelatin in traditional Chinese patent medi-cines by UPLC-QqQ-MS [J ]. Chinese Pharmaceutical Journal ,2023,58 (3):260-265.][ 2 ]谷麟, 程逸凡, 杨光昕, 等. 质谱技术在动物源性食品兽药多残留检测中的研究进展[J ]. 食品与发酵科技,2023,59(2):110-119. [GU Lin, CHENG Yifan,YANG Guangxin, et al. Research progress in the de-tection of veterinary drug residues in food of animal origin by mass spectrometry [J ]. Food and Fermenta-tion Science & Technology ,2023,59 (2):110-119.][ 3 ]郑耀林, 林秋凤, 杨乐, 等. 液相色谱-高分辨质谱技术在兽药残留检测中的研究进展[J ]. 食品科学,2023,44(3):359-365. 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一种具有抗炎活性的辣木叶多糖MLP100-3组分、其纯化工艺及其应用[发明专利]
专利名称:一种具有抗炎活性的辣木叶多糖MLP100-3组分、其纯化工艺及其应用
专利类型:发明专利
发明人:王磊,崔璨,陈晓阳
申请号:CN201810415879.4
申请日:20180503
公开号:CN108409884A
公开日:
20180817
专利内容由知识产权出版社提供
摘要:本发明涉及一种具有抗炎活性的辣木叶多糖MLP100‑3组分。
所述辣木叶多糖MLP100‑3的单糖组成比例为,鼠李糖∶阿拉伯糖∶木糖∶甘露糖∶葡萄糖∶半乳糖的质量比为
1.2‑1.3∶4.5‑5.4∶4.2‑5.5∶3.4‑4.0∶5.6‑6.6∶5.9‑6.5。
本发明还提供了辣木叶多糖MLP100‑3的纯化方法。
本发明所述辣木叶多糖组分MLP100‑3在小鼠腹腔巨噬细胞RAW264.7体外炎症模型中呈现出较好的抑制NO及炎症因子TNF‑α生成的能力,该发明可应用于调节机体免疫功能以及抗炎药物的开发与利用中。
申请人:华南农业大学
地址:510642 广东省广州市天河区五山路483号
国籍:CN
代理机构:北京欣永瑞知识产权代理事务所(普通合伙)
代理人:张庆敏
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常见细菌真菌中英文对照(史上最全,纯手工整理)
DNSN-1_在秀丽隐杆线虫DNA复制起始期间招募GINS_用于CMG解旋酶组装
广东药科大学学报第39卷relationship between community-acquired respiratory distress syndrome toxin and the high-mobility group box protein1-toll-like receptors-myeloid differentiation factor88signaling pathway in mycoplasma pneumoniae pneumoni a[J].Ital J Pediatr,2022,48:64.[11]LUO H,HE J,QIN L,et al.Mycoplasma pneumoniae lipidslicense TLR-4for activation of NLRP3inflammasome and autophagy to evoke a proinflammatory response[J].Clin Exp Immunol,2021,203(1):66-79.[12]LING D Y,ZHAO Y,ZHANG Z W,et al.Morphine inhibitsthe promotion of inflammatory microenvironment on chronic tibial cancer pain through the PI3K-AKt-NF-κB pathwa y[J].Am J Transl Res,2020,12(10):6868.[13]MA J,CHAN C C,HUANG W C,et al.Berberin inhibits pro-inflammatory cytokine-induced IL-6and CCL11production via modulation of STA T6pathway in human bronchial epithelial cell s[J].Int J Med Sci,2020,17(10):1464.[14]ARORA S,AHMAD S,IRSHAD R,et al.TLRs in pulmonarydisease s[J].Life Sci,2019,233:116671.[15]陈金妮,李大伟,林秋玉,等.难治性肺炎患儿感染病原学及HMGB1、TLR2、TLR4表达[J].中华医院感染学杂志, 2020,30(18):2847-2850.[16]黄坚,王伏英.支原体肺炎患儿外周血单核细胞TLR4、IL-6、TNF-α表达变化及临床意义[J].中国妇幼保健,2020,35(2):276-278.[17]刘洁,刘英,杨维民.TLR2/4、MYD88和NF-κB P65在支原体肺炎小鼠中的表达及相关性[J].解剖学研究,2020,42(5): 442-445.[18]宋瑜婷,彭彬,王莹,等.TLR4/MD2/NF-κB信号通路与疾病的研究进展[J].免疫学杂志,2020,36(7):633-638.[19]陈建翠,姜琦.下调IRAK1基因调控NF-κB信号通路抑制宫颈癌细胞生长的机制研究[J].免疫学杂志,2019,35(7): 587-592.[20]黄晗,李凤英,李扬,等.灵芝多糖对脓毒症急性肺损伤大鼠肺功能及TLR4/NF-κB通路存影响[J].中草药,2021,52(8):2351-2355.[21]ZHANG H,LI X,WANG J,et al.Baicalin relieves myco‐plasma pneumoniae infection-induced lung injury through regulating microRNA-221to inhibit the TLR4/NF-κB signaling pathwa y[J].Mol Med Rep,2021,24:571.[22]宋少华,李海舰,李莉娟,等.白芷乙素对溃疡性结肠炎小鼠的影响[J].广东药科大学学报,2021,37(5):100-105.(责任编辑:周鹏)DNSN-1在秀丽隐杆线虫DNA复制起始期间招募GINS用于CMG解旋酶组装CDC45-MCM-GINS解旋酶的组装是真核生物DNA复制起始期间的关键调控步骤。
reads数,测序深度笔记
reads数,测序深度笔记前⾔对于测序深度,reads的计算,以及相关的数据量不太清楚,找了⼀个篇⽂章,源于公众号“嘉因”,后⽅有链接。
Sequencing depth.The amount of sequencing needed for a given sample is determined by the goals of the experiment and the nature of the RNA sample.Experiments whose purpose is to evaluate the similarity between the transcriptional profiles of two polyA+ samples may require only modest depths of sequencing (e.g. 30M pair-end reads of length > 30NT, of which 20-25M are mappable to the genome or known transcriptome, Experiments whose purpose is discovery of novel transcribed elements and strong quantification of known transcript isoforms requires more extensive sequencing. The ability to detect reliably low copy number transcripts/isoforms depends upon the depth of sequencing and on a sufficiently complex library. For experiments from a typical mammalian tissue or in which sensitivity of detection is important, a minimum depth of 100-200 M 2 x 76 bp or longer reads is currently recommended. [Specialized studies in which the prevalence of different RNAs has been intentionally altered (e.g. “normalizing” using DSN) as part of sample preparation need more than the read amounts (>30M paired end reads) used for simple comparison (see above). Reasons for this include: (1) overamplification of inserts as a result of an additional round of PCR after DSN and (2) much more broad coverage given the nature of A(-) and low abundance transcripts.上述⽂字翻译如下:根据研究⽬的决定测序深度:⽬的1:通过抓取polyA尾建库(只测那些带有polyA尾巴的基因,⼤多是蛋⽩编码基因),寻找样品间基因转录谱的相似性,只需要30M reads,长度⼤于30nt即可,双端测序,其中20-25M能够回帖(map)到已知转录组上。
放线菌属新种
Abbreviation : CCUG, Culture Collection of the University of Go $ teborg. The GenBank accession number for the 16S rRNA gene sequence of strain CCUG 41710T is AJ276405. 01587 # 2001 IUMS
Fig. 1. Similarity dendrogram based on whole-cell protein patterns of Actinomyces marimammalium sp. nov. and related species. Levels of correlation are expressed as percentages of similarity for convenience.
International Journal of Systematic and Evolutionary Microbiology (2001), 51, 151–156
Printed in Great Britain
Actinomyces marimammalium sp. nov., from marine mammals
Keywords : taxonomy, phylogeny, Actinomyces marimammalium, 16S rRNA
INTRODUCTION
In recent years, the Gram-positive asporogenous rodshaped organisms of the genus Actinomyces and related taxa have attracted the attention of clinical microbiologists, mainly because of a growing awareness of their presence in clinical specimens and also because of increased taxonomic interest (Funke et al., 1997a). Knowledge of the taxonomic inter-relationships of species of the genus Actinomyces has improved dramatically in recent years particularly with the use of 16S rRNA gene sequencing (Pascual et al., 1997a ; Lawson et al., 1997). Indeed, the application of molecular sequence data in conjunction with improved phenotypic methods of analysis has not only resulted in better species identification but also greatly facilitated the recognition of new taxa, particularly from human clinical sources [e.g. Actinomyces europaeus (Funke et al., 1997b) ; Actinomyces graevenitzii (Pascual et al., 1997b) ; Actinomyces neuii subspp. anitratus and neuii (Funke et al., 1994) ; Actinomyces radingae (Wu $ st et al., 1995) ; Actinomyces turicensis (Wu $ st et al., 1995) ; Actinomyces urogenitalis (Nikolaitchouk et al., 2000) ; Actinobaculum schaalii (Lawson et al., 1997) ;
辣椒中常见产毒性真菌的裸磁珠-多重实时荧光定量PCR检测方法
辣椒中常见产毒性真菌的裸磁珠-多重实时荧光定量PCR检测方法金燕;张薇薇;王溯源;叶正茂;张立实;裴晓方【摘要】Objective To establish a method for detecting 3 common toxigenic molds (Aspergillus, Penicillium, and Fusarium) based on non-modified magnetic beads coupled with multiple real-time PCR (NMB-multiple qPCR). Methods The primers and genus-specific probes were designed based on the rDNA sequences to develop a multiple real-time PCR using non-modified magnetic bead to enrichment of fungal spores. The sensitivity, specificity and repeatability of this assay were evaluated. Results The detection limit of this assay for spiked samples was 104 CFU/g, demonstrating a 10-fold greater detection sensitivity of this assay than that of real-time PCR. The NMB-multiple qPCR assay also showed good specificity and reproducibility and yielded comparable results with those by traditional colony counting method for spiked samples (P>0.05). Conclusion NMB-multiple qPCR assay we established allows rapid and sensitive detection of common mycotoxigenic fungi in paprika.%目的:将多重实时荧光定量PCR技术与裸磁珠富集技术联用,建立辣椒样品中常见3类(曲霉属、青霉属、镰刀菌属)产毒性真菌的快速定量检测方法。
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1077-2626/06/$20.00 ß 2006 IEEE
methods based on the a priori knowledge that the data was produced by a high-order finite element simulation (notable exceptions are Coppola et al. [6] with respect to particle tracking, Wiley et al. [7], [8], [9] with respect to ray-casting of curved-quadratic elements, and Brasher and Haimes [10] with respect to cut-planes). Current visualization methods are typically based on lower-order representations of the data (lower in the sense of what is customarily used in high-order numerical simulations)—for instance, using linear interpolation for determining the values of data in areas not at grid locations. Accurate low-order approximations of high-order data require far more degrees of freedom than the original data set and may introduce data artifacts due to mismatches in the approximation properties of the low-order representation and the original high-order discretization.
IEEE TRANSACTIONS ON VISUALIZATION AND COMPUTER GRAPHICS, VOL. 12, NO. 1, JANUARY/FEBRUARY 2006
1
Ray-Tracing Polymorphic Multidomain Spectral/hp Elements for Isosurface Rendering
Blake Nelson and Robert M. Kirby, Member,pose of this paper is to present a ray-tracing isosurface rendering algorithm for spectral/hp (high-order finite) element methods in which the visualization error is both quantified and minimized. Determination of the ray-isosurface intersection is accomplished by classic polynomial root-finding applied to a polynomial approximation obtained by projecting the finite element solution over element-partitioned segments along the ray. Combining the smoothness properties of spectral/hp elements with classic orthogonal polynomial approximation theory, we devise an adaptive scheme which allows the polynomial approximation along a raysegment to be arbitrarily close to the true solution. The resulting images converge toward a pixel-exact image at a rate far faster than sampling the spectral/hp element solution and applying classic low-order visualization techniques such as marching cubes.
THE use of simulation science [1] as a means of scientific inquiry is increasing at a tremendous rate. The process of mathematically modeling physical phenomena, experimentally estimating important key modeling parameters, numerically approximating the solution of the mathematical model, and computationally solving the resulting algorithm has inundated the scientific and engineering worlds, allowing for rapid advances in our modern understanding and utilization of the world around us.
. The authors are with the School of Computing and Scientific Computing and Imaging Institute, University of Utah, Salt Lake City, UT, 84112. E-mail: {bnelson, kirby}@.
Quantification and, ideally, elimination of error is a critical component of the scientific process; it allows scientists to judiciously evaluate which component of the simulation science process (e.g., modeling, numerical approximation, parameter estimation) requires refinement in light of comparison with the real phenomenon of interest. Over the last 40 years, tremendous effort has been exerted in the pursuit of numerical methods which are both flexible and accurate, hence providing sufficient fidelity to be employed in the numerical solution of a large number of models and sufficient quantification of accuracy to allow researchers to focus their attention on model refinement and uncertainty quantification. High-order finite element methods (a variant of which is the spectral/hp element methods) using either the continuous Galerkin or discontinuous Galerkin formulation have reached a level of sophistication such that they are now commonly applied to a diverse set of real-life engineering problems in computational solid mechanics, fluid mechanics, acoustics, and electromagnetics (e.g., [2], [3], [4], [5]).
In this paper, we seek to develop a ray-tracing isosurface rendering algorithm which exploits the properties of spectral/hp element data [11], and for which the visualization error is both quantified and minimized. This is accomplished by reducing the determination of the rayisosurface intersection of spectral/hp element data to classic polynomial root-finding applied to a polynomial approximation obtained by projecting the finite element solution over element-partitioned segments along the ray. Combining the smoothness properties of spectral/hp finite elements with classic orthogonal polynomial approximation theory, we devise an adaptive scheme which allows the polynomial approximation along a ray-segment to be arbitrary close to the true solution. The methodology presented herein has broader application than merely spectral/hp elements; this methodology can be adapted to any volumetric function which meets the smoothness properties of the spectral/hp element discretizations exploited in this work.