bioactive论文发表

发表代表性论文目录[1] 段金廒*,吴启南,宿树兰,周荣汉.中药资源化学学科的建立与发展[J].中草药, 2012, 43: 1665-1671.[2] 段金廒*.中药资源化学研究技术体系的建立及其应用[J].中国药科大学学报,2012,4:289-292.[3] 段金廒*, 严辉, 宿树兰, 钱大玮, 吴启南, 周荣汉. 药材适宜采收期综合评价模式的建立与实践. 中草药, 2010, 41 (11): 1755-1760.[4] 段金廒*, 宿树兰, 吕洁丽, 严辉, 丁安伟. 药材产地加工传统经验与现代科学认识. 中国中药杂志, 2009, 34（24）: 3151-3157.[5] 段金廒*, 宿树兰, 钱大玮, 唐于平, 周荣汉. 中药资源化学研究思路方法与进展. 中国天然药物, 2009, 7 (5): 333-340.[6] 段金廒*, 肖小河, 宿树兰, 赵润怀, 严辉. 中药材商品规格形成模式的探讨——以当归为例. 中国现代中药, 2009, 11 (6): 14-17.[7] 段金廒*, 周荣汉, 宿树兰, 丁安伟, 吴启南. 我国中药资源科学发展现状及展望. 自然资源学报, 2009, 24 (3): 378-386.[8] 段金廒*, 宿树兰, 严辉, 钱大玮. 药材初加工“杀青”环节与药材品质形成的探讨. 中药材, 2011, 24(1): 1-6.[9] 宿树兰, 段金廒*, 欧阳臻, 郭盛, 刘利. 我国桑属(Morus L.)药用植物资源化学研究进展. 中国现代中药, 2012, 14(7): 1-6.[10] 严辉,段金廒*,王瑞琪,郭增祥,钱大玮,宋秉生,何子清,高鸿英.岷县当归产量预报模型研究[J].中药材,2012,4:511-514.[11] 段金廒*,张伯礼,范欣生,张艳军,高月,林娜,钟赣生.中药配伍禁忌研究思路与技术体系框架[J].世界科学技术(中医药现代化),2012,3:1537-1546.[12] 段金廒*,宿树兰,范欣生,钱大玮,唐于平,尚尔鑫,郭建明,刘培.基于药物相互作用探讨中药七情合和相反/相恶/相畏配伍禁忌作用模式与机制[J].世界科学技术(中医药现代化),2012,3:1547-1552.[13] 陈艳琰,钱大玮,尚尔鑫,钱叶飞,刘培,宿树兰,郭建明,唐于平,段金廒*.基于化学成分相互作用探讨芫花与甘草配伍禁忌的机制[J].药学学报,2012,8:1043-1048.[14] 华永庆,段金廒,钱大玮,江曙,丁爱华,宿树兰,郭建明. 胃肠道系统功能评价技术体系的建立及其在中药配伍禁忌研究中的应用. 世界科学技术—中医药现代化, 2012, 14 (3): 17-25.[15] 钱叶飞,尚尔鑫,段金廒*,钱大玮,唐于平. 基于液质联用数据库技术的中药及天然药物化学成分快速鉴定方法的建立. 中国中药杂志, 2012, 37 (21): 3256-3263.[16] 段金廒*, 吴勉华, 范欣生, 张艳军, 刘成海, 唐于平, 尚尔鑫, 王中越. 中医方药量-效关系科学问题的探讨. 南京中医药大学学报, 2010, 26 (1): 1-6.[17] 宿树兰, 段金廒*, 李文林, 唐于平, 范欣生. 基于物质基础探讨中药“十八反”配伍致毒/增毒机制. 中国实验方剂学杂志, 2010, 16 (1): 123-129.[18] 段金廒*, 宿树兰, 唐于平, 范欣生. 中药药对配伍组合的现代认识. 南京中医药大学学报, 2009, 25 (5): 330-333.[19] 段金廒*, 范欣生, 宿树兰, 唐于平, 尚尔鑫, 李文林, 郭建明, 华永庆. 中药及方剂量效关系的研究进展与思考. 南京中医药大学学报, 2009, 25 (4): 241-245.[20] 段金廒*, 陆茵,陈建伟, 丁安伟, 孙世发, 朱荃, 范欣生. 方剂现代研究的思路与方法. 南京中医药大学学报(自然科学版), 2006, 22 (1): 1-3.[21]Shulan Su, Jin-Ao Duan*, Peijuan Wang, Pei Liu, Jianming Guo, Erxin Shang, DaweiQian, Yuping Tang, Zongxiang Tang. A metabolomic study of biochemical changes in the plasma and urine of primary dysmenorrhea patients using UPLC-MS coupled with pattern recognition approach [J]. Journal of Proteome Research, 2013, 12 (2): 852–865. (IF=5.113) [22]Hongyue Ma, Min Hong, Jinao Duan*, Pei Liu, Xinsheng Fan, Erxin Shang, Shulan Su, Jianming Guo, DaweiQian, Yuping Tang. Altered cytokine gene expression in peripheral blood monocytes across the menstrual cycle in primary dysmenorrhea: a case-control study [J]. PLos One, 2013, 8(2): e55200. (IF=4.092) [23]Pei Liu, Jinao Duan*, Peijuan Wang, DaweiQian, JianmingGuo, Erxin Shang, Shulan Su, Yuping Tang, Zongxiang Tang. Biomarkers of primary dysmenorrhea and herbal formula intervention: an exploratory metabonomics study of blood plasma and urine [J]. Molecular BioSystems, 2013, 9 (1): 77–87. (IF=3.534)[24] Shulan Su, YongqingHua, Yanyan Wang, Wei Gu, Wei Zhou,Jin-ao Duan*, Haifeng Jiang, Ting Chen, Yuping Tang. Evaluation of the anti-inflammatory and analgesic properties of individual andcombined extracts from Commiphoramyrrha, and Boswelliacarterii.Journal of Ethnopharmacology, 2012, 139: 649-656.25] L. Yu, B.P. Jiang,D. Luo, X.C. Shen, S. Guo, J.A. Duan*, Y.P. Tang. Bioactive components in the fruits of Ziziphusjujuba Mill. against theinflammatory irritant action of Euphorbia plants. Phytomedicine,2012, 19 (3-4): 239-244.[26] Li Liu, Jin-aoDuan*, Yuping Tang, JianmingGuo, Nianyun Yang, Hongyue Ma, Xuqin Shi. Taoren-Honghua herb pair and its main components promoting blood circulation through influencing on hemorheology, plasma coagulation and platelet aggregation. Journal of Ethnopharmacology, 2012, 139 (1): 381-387.[27] Hanqing Wang, Sheng Guo, DaweiQian, YefeiQian, Jin-aoDuan. Comparative analysis of quinolizidine alkaloids from different parts of Sophoraalopecuroides seeds by UPLC-MS/MS. Journal of Pharmaceutical and Biomedical Analysis, 2012, 67-68: 16-21.[28] Wei-Wei Tao, Jin-Ao Duan,* Nian-Yun Yang, Sheng Guo, Zhen-Hua Zhu,Yu-Ping Tang and Da-Wei Qian. Determination of Nucleosides andNucleobases in the Pollen of TyphaAngustifolia by UPLC-PDA-MS. Phytochemical Analysis, 2012, 23 (4): 373-378.[29] Rui Liu, Jin-ao Duan*, Sheng Guo, Jian-mingGuo, Da-wei Qian, Yu-ping Tang. Development of a fingerprint method for animal horn classification by liquid chromatography coupled with hierarchical clustering analysis. Journal of Liquid Chromatography & Related Technologies, 2012, 35 (2):205-214.[30] Li Liu, Hongyue Ma, Yuping Tang, Wenxing Chen, Yin Lu, JianmingGuo,Jin-ao Duan*. Discovery of estrogen receptor a modulators from natural compounds in Si-Wu-Tang series decoctions using estrogen-responsive MCF-7 breast cancer cells. Bioorganic & Medicinal Chemistry Letters, 2012, 22 (1): 154-163[31] Pei Liu, Jin-ao Duan*, Jian-mingGuo, Er-Xin Shang, Da-weiQian,Shu-lan Su and Yu-ping Tang. Identification of major chemical constituents and their metabolites in rat plasma and various organs after oral administration of effective Xiang-Fu-Si-Wu Decoction fraction by UPLC-Q-TOF-MS and Metabolynx. Journal of Liquid Chromatography & Related Technologies, 2012, DOI:10.1080/10826076.2012.698676.[32] Wei-Wei Tao, Jin-AoDuan*, Nian-Yun Yang, Yu-Ping Tang, Meng-Zhu Liu, Ye-FeiQian. Antithrombotic phenolic compounds from Glycyrrhiza uralensis.Fitoterapia,2012, 83(2):422-425.[33] G. Yu, Y.X. Ma, J.-A. Duan*, B.S. Song, Z.Q. He. Identification of differentially expressed genes involved in early bolting of Angelica sinensis (Apiaceae). Genetics and Molecular Research, 2012, 11 (1): 494-502.[34] Sheng Guo,Jin-ao Duan*, Yuping Tang, DaweiQian, Zhenhua Zhu, YefeiQian, ErxinShang, Shulan Su. UHPLC-TOFMS coupled with chemometric method as a powerful technique for rapid exploring of differentiating components between two Ziziphus species Journal of Separation Science,2011, 34(6): 659-666.[35] Shulan Su, Tuanjie Wang,Jin-Ao Duan*, Wei Zhou, Yong-Qing Hua, Yu-Ping Tang, Li Yu, Da-Wei Qian. Anti-inflammatory and analgesic activity of different extracts of Commiphoramyrrha. Journal of Ethnopharmacology, 2011, 134 (2): 251-258.[36] Sheng Guo, Jin-ao Duan*, Yu Ping Tang, ShuLan Su, Da Wei Qian. Triterpenoid acids from Ziziphus jujube. Chemistry of Natural Compounds, 2011, 47 (1): 138-139.[37] Rui Liu, Jin-ao Duan*, Min Wang, Erxin Shang, JianmingGuo, Yuping Tang. Analysis of active components of rhinoceros, water buffalo and yak horns using two-dimensional electrophoresis and ethnopharmacological evaluation. Journal of Separation Science,2011, 34 (3): 354-362.[38] Pei Liu, Jin-ao Duan*, Yong qingHua, Yu ping Tang, Xin Yao, Shulan Su. Effects of Xiang-Fu-Si-Wu Decoction and its main components for dysmenorrhea on uterus contraction. Journal of Ethnopharmacology, 2011, 133 (2): 591-597.[39] JianmingGuo, Erxin Shang, Jin-ao Duan*, Yuping Tang, DaweiQian. Determination of ligustilide in the brains of freely moving rats using microdialysis coupled with ultra performance liquid chromatography/mass spectrometry. Fitoterapia, 2011, 82 (3): 441-445.[40] JianmingGuo, CaifuXue, Jin-ao Duan,Er-xin Shang, DaweiQian, Yuping Tang, QiangOuyang, Mei Sha. Fast characterization of constituents in HuangKui capsules using UPLC-QTOF-MS with collision energy and MassFragment software. Chromatographia, 2011, 73 (5-6): 447-456.[41] CaifuXue, JianmingGuo, DaweiQian*, Jin-ao Duan*, Erxin Shang, Yan Shu, Yuwei Lu. Identification of the potential active components of Abelmoschusmanihot in rat blood and kidney tissue by microdialysis combined with ultra-performance liquid chromatography/ quadrupole time-of-flight mass spectrometry. Journal of Chromatography B, 2011, 879 (5-6): 317-325.[42] Shulan Su, JianmingGuo, Jin-ao Duan*, Tuanjie Wang, Erxin Shang, DaweiQian, Yuping Tang, Anwei Ding. Ultra-performance liquid chromatography-tandem mass spectrometry analysis of the bioactive components and their metabolites of ShaofuZhuyu decoction active extract in rat plasma. Journal of Chromatography B, 2010, 878 (3-4): 355–362.[43] Min Zhu, Yuping Tang,Jin-ao Duan*, JianmingGuo, ShengGuo, Shulan Su, Erxin Shang, Dawei Qian, Anwei Ding. Roles of paeoniflorin and senkyunolide I in SiWu decoction on antiplatelet and anticoagulation activitiesJournal of Separation Science,2010, 33 (21), 3335–3340.[44] Li Liu, Hongyue Ma, Nianyun Yang, Yuping Tang, JianmingGuo, Weiwei Tao, Jin-ao Duan*. A series of natural flavonoids as thrombin inhibitors: structure-activity relationships Thrombosis Research, 2010, 126 (5): 365–378.[45] Li Liu, Jin Ao Duan*, Yu Ping Tang, Hong Yue Ma, ShuLan Su, Jian Ming Guo, Yong Qing Hua. The protective effects of the active fraction of ShaofuZhuyu decoction on hydrogen peroxide-induced oxidative injury in vascular smooth muscle cells. Molecules, 2010, 15 (8), 5066–5078.[46] Yuping Tang, Min Zhu, Sheng Yu, YongqingHua, Jin-Ao Duan*, Shulan Su, Xu Zhang, Yin Lu, Anwei Ding. Identification and comparative quantification of bio-active phthalides in essential oils from Si-Wu-Tang, Fo-Shou-San, Radix Angelica and RhizomaChuanxiong. Molecules, 2010, 15(1), 341–351.[47] ShulanSu, YongqingHua, Jin-Ao Duan*, WeiZhou, ErxinShang, YupingTang. Inhibitory Effects of Active Fraction and Its Main Components of ShaofuZhuyu Decoction on Uterus Contraction. American Journal of Chinese Medicine, 2010, 38 (4): 777–787.[48] Wei Zhou, ShuLan Su*, Jin-Ao Duan*, Jian Ming Guo, Da Wei Qian, ErXin Shang, Jin Zhang. Characterization of the active constituents in Shixiao San using bioactivity evaluation followed by UPLC-QTOF and Markerlynx analysis. Molecules, 2010, 15 (9): 6217–6230.[49] Jin-ao Duan*, PengfeiHou, Yuping Tang, Pei Liu, Shulan Su, Hanqing Liu. A new sesquiterpene and other constituents fromSaussurealappa root,Natural Product Communications, 2010, 5 (10): 1531-1534.[50] Sheng Guo, Jin-Ao Duan*, Yu-Ping Tang, Zhen-Hua Zhu, Ye-FeiQian, Nian-Yun Yang, Er-Xin Shang, Da-Wei Qian. Characterization of nucleosides and nucleobases in fruits of Ziziphusjujuba by UPLC-DAD-MS. Journal of Agricultural and Food Chemistry, 2010, 58 (19), 10774–10780.[51] Sheng Guo,Jin-Ao Duan*, Yu-Ping Tang, Nian-Yun Yang, Da-Wei Qian, Shu-Lan Su, Er-Xin Shang. Characterization of triterpenic acids in fruits of Ziziphus species by HPLC-ELSD-MS. Journal of Agricultural and Food Chemistry,2010, 58 (10), 6285–6289.[52] Pei Liu, Li Liu, Yu Ping Tang,Jin Ao Duan*, Nian Yun Yang. A new cerebroside and its anti-proliferation effect on VSMCs from the radix of Cyperusrotundus L. Chinese Chemical Letters, 2010, 21 (5): 606–609.[53] Wei Wei Tao, Nian Yun Yang, Li Liu, Jin-Ao Duan*, De Kang Wu, Da Wei Qian, Yu Ping Tang. Two new cerebrosides from the pollen of Typhaangustifolia. Fitoterapia. 2010, 81 (3): 196–199.[54] Wei Wei Tao, NianYun Yang,Jin-Ao Duan*, De Kang Wu, ErXin Shang, Da Wei Qian, Yu Ping Tang. Two new nonacosanetriols from the pollen of Typhaangustifolia. Chinese Chemical Letters, 2010, 21 (2): 209–212.[55] Ying Chen, Jin-ao Duan*, DaweiQian, JianmingGuo, Bingsheng Song, Ming Yang. Assessment and comparison of immunoregulatory activity of four hydrosoluble fractions of Angelica sinensis in vitro on the peritoneal macrophages in ICR mice. International Immunopharmacology, 2010, 10 (4): 422–430.[56] Rui Liu, Min Wang,Jin-ao Duan*, Jian-mingGuo, Yu-ping Tang. Purification and identification of three novel antioxidant peptides from Cornububali (water buffalo horn). Peptides, 2010, 31 (5): 786–793.[57] Nian-Yun Yang, Wei-Wei Tao, Min Zhu, Jin-Ao Duan*, Jian-Guo Jiang. Two new isopimaranediterpenes from the feces of Trogopterusxanthipes. Fitoterapia, 2010, 81 (5): 381-384.[58] Nian Yun Yang, Jin-Ao Duan*, ErXin Shang, Li Juan Tian. Analysis of Sesquiterpene Lactones in Eupatorium lindleyanum by HPLC-PDA-ESI-MS/MS. Phytochemical Analysis, 2010, 21 (2): 144–149.[59] Nian-Yun Yang, Li Liu, Wei-Wei Tao, Jin-Ao Duan*, Li-Juan Tian. Diterpenoids from Pinusmassoniana resin and their cytotoxicity against A431 and A549 cells. Phytochemistry, 2010, 71 (13): 1528–1533.[60] Nian Yun Yang, Wei Wei Tao, Jin-Ao Duan, Antithrombotic flavonoids from the faeces of Trogopterusxanthipes.Natural Product Research, 2010, 24 (19): 1843–1849.[61] JianmingGuo, Jin-ao Duan*, Yuping Tang, NingJia, Xinghu Li, Ji Zhang. Fast onset of action and the analgesic and sedative efficacy of essential oil from RhizomaChuanxiong after nasal administration. Pharmazie, 2010, 65 (4): 296–299.[62] JianmingGuo, Erxin Shang,Jin-ao Duan*, Yuping Tang, DaweiQian, Shulan Su. Fast and automated characterization of major constituents in rat biofluid after oral administration of Abelmoschusmanihot extract using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry and MetaboLynx. Rapid Communications in Mass Spectrometry, 2010, 24 (4): 443–453.[63] YueGu, DaweiQian*, Jin-ao Duan*, Zhenzhong Wang, JianmingGuo, Yuping Tang, Sheng Guo. Simultaneous determination of seven main alkaloids of Chelidoniummajus L. by ultraperformance LC with photodiode-array detection. Journal of Separation Science, 2010, 33 (8), 1004–1009.[64] Rui Liu, Min Wang, Jin-Ao Duan*. Antipyretic and antioxidant activities of the aqueous extract of Cornububali (Water Buffalo Horn). The American Journal of Chinese Medicine, 2010, 38 (2): 293–306.[65] 朱振华, 尚尔鑫, 段金廒*, 钱大玮, 唐于平. 中药方剂化学成分液质联用分析中标准质谱库的建立和应用. 分析化学, 2010, 38 (11): 1588-1592.[66] Jie Li Lü, Jing Zhao, Jin-Ao Duan*, Hui Yan, Yu Ping Tang, Lai Bin Zhang. Quality evaluation of Angelica sinensisby simultaneous determination of ten compounds usingLC-PDA. Chromatographia, 2009, 70 (3-4): 455-465.[67] Jie Li Lü,Jin Ao Duan*, Yu Ping Tang, Nian Yun Yang, Lai Bin Zhang. Phthalide mono- and dimers from the radix of Angelica sinensis. Biochemical Systematics and Ecology, 2009, 37 (4): 405-411.[68] XiangchunShen, Yuping Tang, Ruihui Yang, Li Yu, Taihui Fang,Jin-ao Duan*. The protective effect of Zizyphus jujube fruit on carbon tetrachloride-induced hepatic injury in mice by anti-oxidative activities. Journal of Ethnopharmacology, 2009, 122 (3): 555–560.[69] Sheng Guo, Jin-ao Duan*, Yuping Tang, Shulan Su, Erxin Shang, Shumao Ni, DaweiQian. High-performance liquid chromatography—two wavelength detection of triterpenoid acids from the fruits of Ziziphusjujuba containing various cultivars in different regions and classification using chemometric analysis. Journal of Pharmaceutical and Biomedical Analysis, 2009, 49 (5): 1296–1302[70] Sheng Guo, Yu Ping Tang, Jin-Ao Duan*, ShuLan Su, An Wei Ding. Two new terpenoids from fruits ofZiziphus jujube. Chinese Chemical Letters, 2009, 20 (2): 197–200.[71] ShuLan Su, Jin-Ao Duan*, Yu Ping Tang, Xu Zhang, Li Yu, FengRong Jiang, Wei Zhou, Dan Luo, An Wei Ding. Isolation and biological activities of neomyrrhaol and other terpenes from the resin of Commiphoramyrrha. Planta Medica, 2009, 75 (4): 351–355.[72] Nian Yun Yang, Wei Wei Tao, Jin-AoDuan*, Jian Ming Guo, Ling Ling Cao. Four new fatty acid esters from the feces of Trogopterusxanthipes. Lipids, 2009, 44 (9): 849–853.[73] Nian Yun Yang, Jin-Ao Duan*, Da Wei Qian. Simultaneous quantification of four sesquiterpene lactones inEupatorium lindleyanum DC. by RP-LC.Chromatographia,2009, 70 (1): 205–209.[74] Nian Yun Yang, Wei Wei Tao, Jin-Ao Duan* Three new lipids from the seeds of Trogopterusxanthipes. Journal of Chemical Research, 2009, (7): 423-426.[75] Nianyun Yang, Weiwei Tao,Jin-ao Duan*. Determination of Biflavones from Trogopterus Feces by HPLC.Journal of Food and Drug Analysis, 2009, 17 (5): 376–379.[76] Nianyun Yang, Weiwei Tao, Jin-ao Duan*. Four new long-chain aliphatics from the feces of Trogopterusxanthipes. Journal of Asian Natural Products Research, 2009, 11 (12): 1302– 1309.[77] JianmingGuo, Jin-ao Duan*, Erxin Shang, Yuping Tang, DaweiQian. Determination of ligustilide in rat brain after nasal administration of essential oil from RhizomaChuanxiong. Fitoterapia,2009, 80 (3): 168–172.[78] Jin-ao Duan*, Liuying Wang, ShihuiQian, ShulanSu, Yuping Tang. A New Cytotoxic PrenylatedDihydrobenzofuran Derivative and Other Chemical Constituents from the Rhizomes of Atractylodeslancea DC. Archieves of Pharmacal Research,2008, 31 (8): 965-969.[79] Shulan Su, YongqingHua, Jin-ao Duan*, Erxin Shang, Yuping Tang, XingjieBao, Yin Lu, Anwei Ding. Hypothesis of active components in volatile oil from a Chinese herbs formulation, ‘Shao-Fu-Zhu-Yu Decoction’, using GC-MS and Chemometrics. Journal of Separation Science,2008, 31 (6): 1085-1091.[80] Shulan Su, Li Yu, YongqingHua,Jin-ao Duan*, Haishan Deng, Yuping Tang, Yin Lu, Anwei Ding. Screening and analysis of potential bioactive components from ShaofuZhuyu decoction, using human umbilical vein endothelial cell extract and high performance liquid chromatography coupled with Q-TOF/MS spectrometry, Biomedical Chromatography,2008, 22 (12): 1385-1392.[81] Jie Li Lü, Jin-Ao Duan*, Yu Ping Tang, Two new ceramides from the radix of Angelica sinensis, Journal of Chemical Research, 2008, (11), 658-661.[82] YongqingHua, Shulan Su, Jin-ao Duan*, Qiujuan Wang, Yin Lu, Long Chen. Danggui–Shaoyao–San, a traditional Chinese prescription, suppresses PGF2a production in endometrial epithelial cells by inhibiting COX-2 expression and activity,Phytomedicine, 2008, 15 (12): 1046-1052.

近些年来，关于外泌体的研究如火如荼。外泌体在免疫中抗原呈递、肿瘤的生长与迁移、组织损伤的修复等生理病理上起着重要的作用。同时，不同细胞分泌的外泌体具有不用的组成成分和功能，可作为疾病诊断的生物标志物。 来自华中科技大学同济医学院附属协和医院的崔老师，对外泌体提取积累了自己的心得，并将自己的研究成果发表在 Bioactive Materials 上。 下文是崔老师的分享。

要进行关于外泌体的研究，提取纯化外泌体一直是大家非常关注的问题。 能否获得较高纯度的足量外泌体，直接关系着后续研究能否开展 。虽然目前仍然没有能同时保证外泌体的含量、纯度和生物活性的提取方法，但是我们可以结合当前的实际情况，对外泌体提取做一些探讨和经验总结。

超速离心（差速离心）法是目前最常用的外泌体提取方法 ，即采用低速离心、高速离心交替进行，可分离到大小相近的囊泡颗粒。超离法因操作简单，获得的囊泡数量较多而广受欢迎。本人近期发表的论文中最后也是采用了这种方法。

具体的步骤大家都比较熟悉，无论是提取血清来源的外泌体，还是细胞条件培养基来源的外泌体 ，大家都习惯于先将血清或者条件培养基收集后保存于-80℃的温度下，等达到一定量后，再批量进行超速离心。

这样的话， 大家会发现一个非常严重的问题，尤其是在拍摄电镜的时候——背景杂质很多，并且很多囊泡的形态不够典型，不是文献中描述的那种典型的双凹圆盘状或者茶托状。

通过几次实验的对比，我大概发现了原因， 主要是由于在冻存收集到培养基的时候，没有进行低速离心和普通高速离心。

低速离心可以去除培养基中的未贴壁细胞、死细胞以及大的细胞残骸，而普通高速离心可以去除其中的大型细胞外囊泡。无论是未贴壁细胞、死细胞、大的细胞残骸，还是大型细胞外囊泡，在-80℃冻存及复温的过程中，都会发生破裂，产生小型的细胞膜碎片结构。

这样的话，就导致了电镜背景很杂乱，难以达到高水平论文杂志期刊的要求。另外，还会导致这样一个矛盾现象，即蛋白定量中计算的外泌体产量虚高，而蛋白免疫印迹中外泌体标记物蛋白的含量却不高。

因此，在这里推荐给大家一个小贴士

就是在收集准备提取外泌体的条件培养基或者血清时，一定要提前做到低速离心和普通高速离心这一步，尤其是为了做透射电镜或者蛋白免疫印迹实验而提取的外泌体 ，一定要注意这一点。尽管过程比较费时，但是能生产出有用的结果才是更重要的。

至于其他的外泌体提取方法，如密度梯度离心、色谱柱、超滤离心法、微流控芯片法、磁珠免疫法、多聚物沉淀法，我看到学校里面鲜少有人去尝试，毕竟更麻烦。但不同的方法各有优劣，大家可根据自己的实验，谨慎选用。