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羊膜干细胞分离方法及其细胞学特性鉴定

透析与人工器官 2009年第1期20卷 1-5页

作者：朴正福张海燕丁淑芹小林护佐佐木幸司菊池爱子加茂功樱川宣男李宁首都医科大学附属北京佑安医院肝炎研究所北京100069 北里大学医疗卫生学部

目的按照Kobayashi方法从羊膜间充质细胞(AMCs)的侧群细胞(SP)中分离羊膜干细胞并予以细胞学特性鉴定。方法按文献法取剖腹产手术胎盘,用色素染色法和流式细胞分选技术从AMCs中分离羊膜干细胞进行原代培养,用PI染色DNA分析细胞周期,软... 详细信息

目的按照Kobayashi方法从羊膜间充质细胞(AMCs)的侧群细胞(SP)中分离羊膜干细胞并予以细胞学特性鉴定。方法按文献法取剖腹产手术胎盘,用色素染色法和流式细胞分选技术从AMCs中分离羊膜干细胞进行原代培养,用PI染色DNA分析细胞周期,软琼脂培养法观察细胞体外集落,并与人肝癌细胞系HepG2比较。结果1.从AMCs中成功分离的(羊膜干细胞侧群细胞)AMCs-SP约占总AMCs的0.2%;2.细胞周期结果为AMC-SP细胞增殖缓慢或处于静止期;3.体外培养时细胞群体培增数达40以上,在软琼脂中AMC-SP不形成集落,与人肝癌细胞系HepG2可形成较大集落形成反差。结论AMC-SP具有细胞来源充足、稳定性好、安全性高、无伦理学争议,异体移植无急性免疫排外反应,无致瘤性的优点,可做为再生医学及急性损伤期疾病治疗的重要的人细胞来源应用于临床。

关键词：干细胞羊膜间充质细胞侧群细胞细胞治疗

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一种新型医用动态空气消毒机对MTB等病原微生物杀菌效果的研究

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中国防痨杂志 2018年第4期40卷 411-415页

作者：刘志永王帅张天宇土井教生谭守勇谢小保高亚敏唐运祥姜火风谢武中国科学院广州生物医药与健康研究院呼吸疾病国家重点实验室 510530 日本国家结核预防会结核病研究所广州市胸科医院广东省微生物研究所深圳市达柏嘉生物科技有限公司

目的评价采用新型角柱状光触媒系统的医用空气消毒机（简称“新型空气消毒机”）在人机共存环境状态下杀菌的效果，研究提高杀灭空气中病原微生物，特别是杀灭结核分枝杆菌（MTB;本实验采用Mycobacteriumbovis BCG菌株，即牛型结核分枝... 详细信息

目的评价采用新型角柱状光触媒系统的医用空气消毒机（简称“新型空气消毒机”）在人机共存环境状态下杀菌的效果，研究提高杀灭空气中病原微生物，特别是杀灭结核分枝杆菌（MTB;本实验采用Mycobacteriumbovis BCG菌株，即牛型结核分枝杆菌减毒株，简称“BCG”）的空气消毒技术和方法。方法采用新型空气消毒机，在体积47m^3实验房中对空气中的自然菌进行杀菌试验；设置3个采样点，试验重复3次，每次试验结果取3个采样点检测的菌落数均值，分别计算其消亡率。在相邻2个体积为30m^3的气雾室（一个用于试验对照，计算细菌的自然消亡率；一个用于消毒试验，计算对细菌的杀灭率），对空气中的白色葡萄球菌、BCG进行杀菌试验；试验重复3次，每次均分别计算其自然消亡率和杀灭率。白色葡萄球菌及BCG杀菌试验气雾室内空气中菌落数应分别达到115～5×10^5CFU／m3和3×10^4～8×10^4CFU／m^3。结果空气模拟现场消毒试验采用新型空气消毒机进行空气现场白色葡萄球菌消毒试验，重复3次对照试验空气中白色葡萄球菌自然消亡率分别为20．45％、20．00％、20．36％；重复3次消毒试验杀菌率分别为99．97％、99．96％、99．95％；空气现场自然菌消毒试验重复3次试验空气中自然菌的消亡率分别为93．95％、94．27％、94．32％；空气模拟现场BCG消毒试验，重复3次对照试验空气中BCG自然消亡率分别为35．71％、37．93％、40．58％，重复3次消毒试验杀菌率分别为99．67％、99．61％、99．73％。结论采用角柱状光触媒系统的新型医用空气消毒机对空气环境中的病原微生物及MTB有强烈的杀灭作用。

关键词：分枝杆菌结核消毒净化卫生用具和设备空气微生物学评价研究

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Real-world Effectiveness and Tolerability of Interferonfree Direct-acting Antiviral for 15,849 Patients with Chronic Hepatitis C:A Multinational Cohort Study

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Journal of Clinical and Translational Hepatology 2024年第7期12卷 646-658页

作者： Fanpu Ji Sally Tran Eiichi Ogawa Chung-Feng Huang Takanori Suzuki Yu Jun Wong Hidenori Toyoda Dae Won Jun Liu Li Haruki Uojima Akito Nozaki Makoto Chuma Cheng-Hao Tseng Yao-Chun Hsu Masatoshi Ishigami Takashi Honda Masanori Atsukawa Hiroaki Haga Masaru Enomoto Huy Trinh Carmen Monica Preda Phillip Vutien Charles Landis Dong Hyun Lee Tsunamasa Watanabe Hirokazu Takahashi Hiroshi Abe Akira Asai Yuichiro Eguchi Jie Li Xiaozhong Wang Jia Li Junping Liu Jing Liang Carla Pui-Mei Lam Rui Huang Qing Ye Hongying Pan Jiajie Zhang Dachuan Cai Qi Wang Daniel Q.Huang Grace Wong Vincent Wai-Sun Wong Junyi Li Son Do Norihiro Furusyo Makoto Nakamuta Hideyuki Nomura Eiji Kajiwara Eileen L.Yoon Sang Bong Ahn Koichi Azuma Kazufumi Dohmen Jihyun An Do Seon Song Hyun Chin Cho Akira Kawano Toshimasa Koyanagi Aritsune Ooho Takeaki Satoh Kazuhiro Takahashi Ming-Lun Yeh Pei-Chien Tsai Satoshi Yasuda Yunyu Zhao Yishan Liu Tomomi Okubo norio Itokawa Mi Jung Jun Toru Ishikawa Koichi Takaguchi Tomonori Senoh Mingyuan Zhang Changqing Zhao Raluca Ioana Alecu Wei Xuan Tay Pooja Devan Joanne Kimiko Liu Ritsuzo Kozuka Elena Vargas-Accarino Ai-Thien Do Mayumi Maeda Wan-Long Chuang Jee-Fu Huang Chia-Yen Dai Ramsey Cheung Maria Buti Junqi Niu Wen Xie Hong Ren Seng Gee Lim Chao Wu Man-Fung Yuen Jia Shang Qiang Zhu Yoshiyuki Ueno Yasuhito Tanaka Jun Hayashi Ming-Lung Yu Mindie H.Nguyen Department of Infectious Diseases The Second Affiliated Hospital of Xi’an Jiaotong UniversityXi’anShaanxiChina Division of Gastroenterology and Hepatology Department of MedicineStanford University Medical CenterPalo AltoCAUSA Department of General Internal Medicine Kyushu University HospitalFukuokaJapan Hepatobiliary Division Department of Internal MedicineKaohsiung Medical University HospitalKaohsiung Hepatitis Research Center College of Medicine and Center for Liquid Biopsy and Cohort ResearchKaohsiung Medical UniversityKaohsiung Department of Gastroenterology and Metabolism Nagoya City University Graduate School of Medical SciencesNagoyaJapan Gastroenterology&Hepatology Changi General HospitalSinghealthSingapore Singhealth Duke-NUS Medicine Academic Clinical Program Singapore Department of Gastroenterology Ogaki Municipal HospitalOgakiJapan Department of Internal Medicine Hanyang UniversityCollege of MedicineSeoulKorea Hanyang Institute of Bioscience and Biotechnology Hanyang UniversitySeoulKorea Department of Hepatology The Third People's Hospital of Kunming CityKunmingYunnanChina Department of Gastroenterology Internal MedicineKitasato University School of MedicineSagamiharaJapan Gastroenterological Center Yokohama City University Medical CenterYokohamaJapan Division of Gastroenterology and Hepatology Department of Internal MedicineE-Da Cancer HospitalKaohsiung Department of Gastroenterology and Hepatology Nagoya University Graduate School of MedicineNagoyaJapan Division of Gastroenterology and Hepatology Nippon Medical SchoolTokyoJapan Department of Gastroenterology Yamagata University Faculty of MedicineYamagataJapan Department of Transfusion Medicine and Department of Hepatology Osaka Metropolitan University Graduate School of MedicineOsakaJapan San Jose Gastroenterology San JoseCAUSA Gastroenterology&Hepatology Department Clinic Fundeni InstituteBucharestRomania Division of Gastroenterology and Hepatology University of WashingtonSeattleWAUSA Department of

Background and Aims:As practice patterns and hepatitis C virus(HCV)genotypes(GT)vary geographically,a global real-world study from both East and West covering all GTs can help inform practice policy toward the 2030 HCV elimination *** study aimed to assess the effectiveness and tolerability of DAA treatment in routine clinical practice in a multinational cohort for patients infected with all HCV GTs,focusing on GT3 and ***:We analyzed the sustained virological response(SVR12)of 15,849 chronic hepatitis C patients from 39 Real-World Evidence from the Asia Liver Consortium for HCV clinical sites in Asia Pacific,North America,and Europe between 07/01/2014–07/01/***:The mean age was 62±13 years,with 49.6%*** demographic breakdown was 91.1%Asian(52.9%Japanese,25.7%Chinese/Taiwan residents,5.4%Korean,3.3%Malaysian,and 2.9%Vietnamese),6.4%White,1.3%Hispanic/Latino,and 1%Black/***,34.8%had cirrhosis,8.6%had hepatocellular carcinoma(HCC),and 24.9%were treatment-experienced(20.7%with interferon,4.3%with direct-acting antivirals).The largest group was GT1(10,246[64.6%]),followed by GT2(3,686[23.2%]),GT3(1,151[7.2%]),GT6(457[2.8%]),GT4(47[0.3%]),GT5(1[0.006%]),and untyped GTs(261[1.6%]).The overall SVR12 was 96.9%,with rates over 95%for GT1/2/3/6 but 91.5%for ***12 for GT3 was 95.1%overall,98.2%for GT3a,and 94.0%for ***12 was 98.3%overall for GT6,lower for patients with cirrhosis and treatment-experienced(TE)(93.8%)but≥97.5%for tretment-naive patients regardless of cirrhosis *** multivariable analysis,advanced age,prior treatment failure,cirrhosis,active HCC,and GT3/4 were independent predictors of lower SVR12,while being Asian was a significant predictor of achieving ***:In this diverse multinational realworld cohort of patients with various GTs,the overall cure rate was 96.9%,despite large numbers of patients with cirrhosis,HCC,TE,and GT3/***12 for GT3/6 with cirrhosis and TE was lower but st

关键词： Hepatitis C virus Liver cirrhosis Hepatocellular carcinoma Genotype DAA REAL-C

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人羊膜间充质干细胞的分离及分化潜能的研究

生物医学工程与临床 2010年第1期14卷 15-19页

作者：朴正福丁淑芹张海燕小林护加茂功樱川宣男李宁首都医科大学附属北京佑安医院北京市卫生局肝炎研究所日本生物资源应用研究所

目的建立人羊膜间充质干细胞(hAMC)的分离培养方法及探讨其在再生医学中的潜在应用性。方法取38～40周孕妇剖宫产术后人羊膜,用多种酶消化法制备hAMC悬浮液,并用免疫细胞化学方法探讨其多分化潜能如hAMC的神经及肝脏发生相关蛋白的表达... 详细信息

目的建立人羊膜间充质干细胞(hAMC)的分离培养方法及探讨其在再生医学中的潜在应用性。方法取38～40周孕妇剖宫产术后人羊膜,用多种酶消化法制备hAMC悬浮液,并用免疫细胞化学方法探讨其多分化潜能如hAMC的神经及肝脏发生相关蛋白的表达;荧光激活细胞仪分离法(FACS)检测主要组织相容性复合体(MHC)家族分子,同时探讨其致瘤性。结果从人羊膜中分离出高纯度hAMC,并摸索出其最适合的培养条件;细胞成细长梭形或星形,胞质丰富,细胞核呈圆形,1～3个核仁,易贴壁,生长繁殖快,在体外可连续培养20代以上。免疫细胞化学结果显示:神经发生相关蛋白(Musashi-1、Nestin、Vimentin)阳性及肝脏发生相关蛋白[Flt-1、Integrinβ1、HNF(3β、4α、1α)、C/EBPα、Albumin、AAT]等阳性;人MHC ClassⅡ阴性,人MHC ClassⅠ弱阳性;癌化实验显示hAMC无致瘤性。结论人羊膜中分离提纯的hAMC中至少包含向神经及肝脏细胞分化的干细胞样的细胞群,具有低免疫原性、无致瘤性等特点。这些结果支持hAMC作为新的、安全的细胞来源可用于再生医学研究。

关键词：羊膜羊膜间充质干细胞生物学特性再生医学