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| 1 | Mesenchymal stem cells alleviate TNBS-induced colitis by modulating inflammatory and autoimmune responses显示文摘AIM: To investigate the potential therapeutic effects of mesenchymal stem cells (MSCs) in inflammatory bowel disease (IBD), we transplanted MSCs into an experimental model of IBD. METHODS: A rectal enema of trinitrobenzene sulfonic acid (TNBS) (100 mg/kg body weight) was administered to female BALB/c mice. Bone marrow mesenchymal stem cells (BMSCs) were derived from male green fluorescent protein (GFP) transgenic mice and were transplanted intravenously into the experimental animals after disease onset. Clinical activity scores and histological changes were evaluated. GFP and Sex determining region Y gene (SRY ) expression were used for cell tracking. Ki67 positive cells and Lgr5-expressing cells were determined to measure proliferative activity. Inflammatory response was determined by mea-suring the levels of different inflammatory mediators in the colon and serum. The inflammatory cytokines included tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-2 (IL-2), IL-6, IL-17, IL-4, IL-10, and transforming growth factor (TGF-β). Master regulators of Th1 cells (T-box expressed in T cells, T-bet), Th17 cells (retinoid related orphan receptor gamma(t), RORγt), Th2 cells (GATA family of transcription factors 3, GATA3) and regulatory T cells (forkhead box P3, Foxp3) were also determined. RESULTS: Systemic infusion of GFP-BMSCs ameliorated the clinical and histopathologic severity of colitis, including body weight loss, diarrhea and inflammation, and increased survival (P < 0.05). The cell tracking study showed that MSCs homed to the injured colon. MSCs promoted proliferation of intestinal epithelial cells and differentiation of intestinal stem cells (P < 0.01). This therapeutic effect was mainly mediated by downregulation of both Th1-Th17-driven autoimmune and inflammatory responses (IL-2, TNF-α, IFN-γ, T-bet; IL-6, IL-17, RORγt), and by up-regulation of Th2 activities (IL-4, IL-10, GATA-3) (P < 0.05). MSCs also induced activated CD4 + CD25 + Foxp3 + regulatory T cells (TGF-β, IL-10, Foxp3) with a suppressive capacity on Th1-Th17 effecter responses and promoted Th2 differentiation in vivo (P < 0.05). CONCLUSION: MSCs are key regulators of immune and inflammatory responses and may be an attractive candidate for cell-based therapy of IBD. | Qian-Qian Chen Li Yan Chang-Zheng Wang Wei-Hua Wang Hui Shi Bin-Bin Su Qing-Huan Zeng Hai-Tao Du Jun Wan | 2013 | World Journal of Gastroenterology2013,19,29: | 25 |
| 2 | Secretion of immunoregulatory cytokines by mesenchymal stem cells显示文摘According to the minimal criteria of the International Society of Cellular Therapy, mesenchymal stem cells(MSCs) are a population of undifferentiated cells defined by their ability to adhere to plastic surfaces when cultured under standard conditions, express a certain panel of phenotypic markers and can differentiate into osteogenic, chondrogenic and adipogenic lineages when cultured in specific inducing media. In parallel with their major role as undifferentiated cell reserves, MSCs have immunomodulatory functions which are exerted by direct cell-to-cell contacts, secretion of cytokines and/or by a combination of both mechanisms. There are no convincing data about a principal difference in the profile of cytokines secreted by MSCs isolated from different tissue sources, although some papers report some quantitative but not qualitative differences in cytokine secretion. The present review focuses on the basic cytokines secreted by MSCs as described in the literature by which the MSCs exert immunodulatory effects. It should be pointed out that MSCs themselves are objects of cytokine regulation. Hypothetical mechanisms by which the MSCs exert their immunoregulatory effects are also discussed in this review. These mechanisms may either influence the target immune cells directly or indirectly by affecting the activities of predominantly dendritic cells. Chemokines are also discussed as participants in this process by recruiting cells of the immune systems and thus making them targets of immunosuppression. This review aims to present and discuss the published data and the personal experience of the authors regarding cytokines secreted by MSCs and their effects on the cells of the immune system. | Dobroslav Kyurkchiev Ivan Bochev Ekaterina Ivanova-Todorova Milena Mourdjeva Tsvetelina Oreshkova Kalina Belemezova Stanimir Kyurkchiev | 2014 | World Journal of Stem Cells2014,6,5: | 20 |
| 3 | 间充质干细胞移植治疗急性肺损伤的研究进展显示文摘急性肺损伤是一种临床常见的危重病症,临床上传统的治疗方法一般以尽早去除诱因、控制感染、机械通气及器官功能支持治疗为主。间充质干细胞属于成体干细胞的一种,能主动归巢至肺损伤部位,并通过向肺泡和支气管上皮细胞分化参与组织修复,同时间充质干细胞能够调节急性肺损伤时局部和全身炎症反应和免疫紊乱,从而发挥治疗作用,可能是治疗急性肺损伤的一个很有前景的方法。作者就间充质干细胞移植治疗急性肺损伤的研究进展进行综述。 | 强勇 梁贵友 | 2014 | 中华细胞与干细胞杂志(电子版)2014,4,3: | 6 |
| 4 | 自体外周血干细胞移植对乙型肝炎肝硬化失代偿期患者肝脏声脉冲辐射力成像的影响显示文摘目的观察自体外周血干细胞(APBSC)移植对乙型肝炎肝硬化失代偿期患者肝脏声脉冲辐射力成像值(ARFI)的影响。方法采用前瞻性随机对照的方法,将68例入组患者随机分组,33例为治疗组,综合内科治疗基础上接受APBSC移植治疗,35例为对照组,单纯内科综合治疗。观察患者移植后12周、24周、36周、48周血总胆红素、凝血酶原时间(PT)、白蛋白(AN)水平和脾脏大小、肝脏ARFI等改变,并与移植治疗前及同期对照组比较。据资料不同分别用t检验、非参数检验或χ^2检验。结果治疗组患者治疗后24周、36周、48周Alb、PT改善,36周、48周脾脏大小、肝脏ARFI指数与治疗前及对照组比较改善。移植后36周、48周脾脏长度分别为(140.09±23.05)mm、(139.55±24.12)mm,与移植前[(149.36±24.56)mm]差异有统计学意义(t值分别为3.43、-2.21,P〈0.05),与同期对照组[(150.26±18.27)mm、(151.66±19.76)mm]比较,差异有统计学意义(t值分别为-2.02、-2.27,P〈0.051。移植后36周、48周ARFI分别为(2.21±0.43)m/s、(2.19±0.43)m/s,与移植前[(2.49±0.31)m/s]差异有统计学意义(t=3.00,P=0.005;t=3.21,P=0.003),与同期对照组比较差异有统计学意义(P〈0.05)。结论APBSC移植治疗能够改善乙型肝炎肝硬化失代偿期患者的肝功能,肝脏ARFI指数及肝脏纤维化程度。 | 邓勤智 蔡挺 张顺 胡爱荣 张行芬 黄建荣 | 2015 | 中华肝脏病杂志2015,23,11: | 6 |
| 5 | 间充质干细胞的肝细胞分化及其治疗终末期肝病的临床研究进展显示文摘间充质干细胞(mesenchymal stem cells,MSCs)是一类具有自我更新和多向分化能力的多能干细胞,研究发现其具有多种表面抗原,但无特异性, | 何宏亮 高志良 | 2014 | 中华肝脏病杂志2014,22,5: | 5 |
| 6 | Application of human bone marrow-derived mesenchymal stem cells in the treatment of radiation-induced gastrointestinal syndrome显示文摘Nuclear accidents and terrorism present a serious threat for mass casualty.Accidental or intended radiation exposure leads to radiation-induced gastrointestinal(GI)syndrome.However,currently there are no approved medical countermeasures for GI syndrome.Thus,developing novel treatments for GI syndrome is urgent.Mesenchymal stem cells(MSCs)derived from bone marrow are a subset of multipotent adult somatic stem cells that have the ability to undergo self-renewal,proliferation and pluripotent differentiation.MSCs have advantages over other stem cells;they can be easily isolated from patients or donors,readily expanded ex vivo,and they possess reparative and immunomodulatory properties.Moreover,MSCs have been shown to be powerful tools in gene therapy and can be effectively transduced with vectors containing therapeutic genes.Therefore,the therapeutic potential of MSCs has been brought into the spotlight for the clinical treatment of GI syndrome.In this review,we discuss the possible role of MSCs in radiation-induced GI syndrome. | YANG Chao DAI Wei Min CHEN Hai Xu WU Ben Yan | 2014 | Science China(Life Sciences)2014,57,12: | 4 |
| 7 | TGF-β1和IL-10对间充质干细胞来源细胞外囊泡免疫调节功能的影响显示文摘目的:探讨TGF-β1和IL-10对间充质干细胞(mesenchymal stem cells,MSC)分泌的细胞外囊泡(extracellular vesicle, EV)免疫调控功能的影响。方法:分离扩增人脐带来源的MSC,分别用TGF-β1和IL-10处理72 h,提取上清中MSC-EV。应用BCA法测定提取EV的总蛋白含量,在透射电子显微镜下观察EV的形态结构,流式细胞术检测EV表面标志物以鉴定MSC-EV。在ConA刺激后的人外周血单个核细胞(PBMNC)培养体系中,加入不同处理方法获得的EV,培养72 h后应用流式细胞仪检测PBMNC的凋亡,以及CD4+CD25+/CD127-调节T(Treg)细胞比例。应用CBA试剂盒和ELISA试剂盒检测培养PBMNC上清以及MSC-EV中IL2,IL4,IL6,IL10,IFN-γ,TNF-α,Th17A和TGF-β1的含量。结果:不同处理后的EV在透射电子显微镜下均呈现特征性杯口状膜样结构,均表达CD9,CD44,CD63及CD81。TGF-β1处理后MSC-EV促PBMNC凋亡的能力明显增强(P <0.01),不同处理后EV均可以提高Treg细胞比例。MSC-EV可以提高PBMNC培养上清中IL-10的含量,TGF-β1处理MSC-EV共培养后的PBMNC上清TGF-β1含量较未处理组明显降低(P <0.05)。TGF-β1处理后的MSC-EV样本IL-6含量显著升高,TGF-β1含量降低。结论:TGF-β1处理后MSC-EV的免疫调控功能发生改变,其意义值得进一步研究阐明。 | 马聪 张青宜 郭子宽 王恒湘 | 2018 | 中国实验血液学杂志2018,26,6: | 4 |
| 8 | 脓毒症治疗的新希望:干细胞疗法显示文摘脓毒症(sepsis)是指由感染引起的全身炎症反应综合征,是临床危重患者的主要死亡原因之一。严重的免疫系统功能紊乱是脓毒症患者的共同病理生理学改变。间充质干细胞(menchymal stem cells,MSCs)具有良好的免疫调节作用,可增加抗炎因子IL-10水平、促进前列腺素E2(PGE2)、吲哚胺2,3双加氧酶(IDO)等细胞因子的分泌,调节单核巨噬细胞、T细胞、B细胞、NK细胞等免疫细胞的增殖、分化和功能,可能为脓毒症的治疗提供新的思路与方法。 | 赵尚平 杨明施 | 2013 | 中南大学学报(医学版)2013,38,7: | 4 |
| 9 | 成骨细胞分化、骨形成与修复中转录因子Osx和Satb2的调控作用显示文摘背景:成骨细胞主要来自于骨髓细胞向骨基质中的间充质细胞,一些转录因子或局部因素可促进骨髓基质细胞调节分化为成骨细胞。目的:明确C_2C_12细胞以及Osx与Satb2在骨质疏松修复过程中的作用。方法:取野生型SD大鼠20只,分为正常组10只,假手术组和骨质疏松模型组(模型组)各5只,同时取Osx-KO大鼠10只。模型组和Osx-KO大鼠切除双侧卵巢构建野生型大鼠与Osx-KO大鼠进行骨质疏松模型;假手术组找出双侧卵巢但不切除。检测各组大鼠术后体质量的变化及股骨骨密度含量。体外培养C_2C_12细胞,并设计了si RNA-Satb2、si RNA-Osx,通过细胞实验、基因沉默、western blot法,观察成骨细胞分化的相关Osx与Satb2的表达及对骨质疏松的影响。结果与结论:(1)体质量:造模12周后,模型组、Osx-KO组大鼠较正常组和假手术组大鼠显著增加(P<0.01)。(2)骨密度:模型组、Osx-KO组较正常组和假手术组显著降低(P<0.01)。(3)转录因子:野生型大鼠各因子均有表达,在Osx-KO大鼠中,Satb2、Osx基因的表达显著降低(P<0.001),而Runx2基因在两种类型的大鼠中的表达差异无显著性意义。(4)当对基因进行沉默后:再检测相关的成骨基因发现Satb2、Osx、Runx2、ALP基因的表达水平均有显著的抑制。(5)结果说明:在骨质疏松发病中转录因子Osx、Satb2可能是保护性分子,对成骨细胞分化、骨形成与修复有着关键的调控作用。 | 侯秋科 黄永铨 李昀骏 陈东风 | 2016 | 中国组织工程研究2016,20,7: | 3 |
| 10 | 间充质干细胞免疫调节作用的研究进展显示文摘间充质干细胞(MSC)是一类具有自我更新、增殖和多向分化潜能的干细胞。研究表明,MSC具有低免疫源性,并且可以通过与细胞间直接接触或分泌细胞因子等方式发挥免疫调节作用。当前,MSC在治疗免疫相关性疾病中的研究是一个热点。为了更好了解MSC免疫方面的特性,本文就MSC的生物学特性和免疫调节作用作一综述。 | 陈强星 张剑 | 2016 | 器官移植2016,7,6: | 3 |
| 11 | 骨髓间充质干细胞分泌因子对Ⅰ型变态反应的影响显示文摘目的探讨骨髓间充质干细胞(BMSCs)分泌因子对Ⅰ型变态反应的影响。方法采用全骨髓贴壁法分离和纯化大鼠BMSCs,收集培养第3代的BMSCs上清液获取干细胞分泌因子。制备大鼠抗卵蛋白血清;分离培养大鼠腹腔肥大细胞,用大鼠抗卵蛋白血清特异性致敏肥大细胞,甲苯胺蓝染色法观察干细胞分泌因子对肥大细胞脱颗粒的影响;制备大鼠同种被动皮肤过敏动物模型(PCA),观察干细胞分泌因子对其影响。结果干细胞分泌因子和酮替芬对特异性致敏肥大细胞脱颗粒和大鼠PCA均有明显抑制作用,与模型组相比差异有显著性(F=219.79~357.32,q=21.26~34.05,P<0.01)。结论干细胞分泌的细胞因子对Ⅰ型变态反应有抑制作用。 | 高明敏 刘晓萍 于业军 王家辉 贾跃伟 李薇 | 2012 | 青岛大学医学院学报2012,48,3: | 3 |
| 12 | 间充质干细胞移植后在体内的归巢显示文摘间充质干细胞(MSCs)可由很多组织分离得到,同时也可分化成不同的组织。MSCs具有一些内在特性,如免疫抑制、组织修复、促进造血重建及减轻移植物抗宿主病(GVHD)等,这些特性使得MSCs成为研究的焦点。然而,MSCs输入动物体内后,在体内的迁移归巢、存活情况以及影响因素均不明确。该文归纳总结了相关文献后发现,影响MSCs在各组织归巢的因素很多,包括肺的滤筛作用、MSCs的数量、培养条件及病理状态等,其中病理状态(即组织损伤程度)具有关键作用。MSCs可特异性地归巢到组织损伤部位,其迁移方向遵循趋化因子浓度梯度,这一过程包括多种配体及其受体的参与。深入了解MSCs移植后在体内的归巢过程将为其在临床的应用提供理论依据。 | 施小凤 朱彦 | 2012 | 上海交通大学学报(医学版)2012,32,1: | 3 |
| 13 | 间充质干细胞在炎性关节炎中的应用前景显示文摘间充质干细胞(mesenchymal stem cells,MSCs)是存在于健康人体骨髓、脂肪、脐带、脐血、胎盘等组织中具有多向分化潜能的非造血干细胞,在适宜条件下可诱导分化为软骨细胞、成骨细胞、脂肪细胞、心肌细胞、神经细胞等。近年研究发现MSCs不仅具有多向分化潜力,同时具有强大的免疫调节和抗炎特性,这一特性给多种自身免疫性疾病的治疗带来了希望。本文就MSCs在炎性关节炎中的应用前景做一综述。 | 黄志芳 黄烽 | 2013 | 中华风湿病学杂志2013,17,9: | 2 |
| 14 | The potential and challenges of using stem cells for cardiovascular repair and regeneration显示文摘Recent progress in using stem cells for tissue repair and functional restoration has aroused much attention due to its potential to provide a cue for many diseases such as myocardial infarction.Stem cell therapy for cardiovascular disease has been studied extensively at both experimental and clinical levels.Pluripotent stem cells and mesenchymal stem cells were proven to be effective for myocardial regeneration,angiogenesis,and cardiac functional restoration.In this review,we will concisely discuss advantages and disadvantages of currently-used stem cells for cardiovascular repair and regeneration.The limitations and uniqueness of some types of stem cells will also be discussed.Although substantial progress has been made over the last decade about stem cells in cardiovascular regeneration,many challenges lie ahead before the therapeutic potentials of stem cells can be fully recognized. | Qisi Sun Zhonge Zhang Zhongjie Sun | 2014 | Genes & Diseases2014,1,1: | 2 |
| 15 | TLR1/2通路活化对脐带间充质干细胞免疫状态影响的研究显示文摘目的研究Toll样受体1/2(Toll like receptor 1/2,TLR1/2)活化后是否改变脐带间充质干细胞(umbilical cord mesenchymal stem cell,UCMSC)的免疫状态。方法UCMSC与外周血单个核细胞(peripheral blood mononuclear cell,PBMC)共培养体系中加入TLR1/2激动剂,用流式细胞术检测PBMC的增殖,并检测PBMC对UCMSC的杀伤作用;检测TLR1/2通路活化后UCMSC表面共刺激分子和干细胞标志物的表达变化;荧光定量PCR检测UCMSC多个免疫相关因子表达变化;对UCMSC进行定向诱导分化,检测TLR1/2活化对UCMSC分化能力的影响。结果 TLR1/2通路活化可促进PBMC增殖,并增强PBMC对UCMSC的免疫杀伤效应,但对UCMSC表面共刺激分子〔CD80/CD86/人类白细胞抗原(HLA)-E〕和干细胞标志物(CD29/CD59/CD90)无影响;荧光定量PCR结果表明UCMSC中多个免疫相关因子被TLR1/2明显诱导〔白细胞介素(IL-2,IL-6,IL-10,IL-12),干扰素-γ(IFN-γ),核因子-κB(NF-κB),肿瘤坏死因子-α(TNF-α)〕;TLR1/2不影响UCMSC的定向分化能力。结论 TLR1/2可改变UCMSC的免疫状态,在一定程度诱导针对UCMSC的免疫攻击。 | 马宇 张立 江婷 刘志强 李全生 | 2015 | 四川大学学报(医学版)2015,46,5: | 2 |
| 16 | 骨髓间充质干细胞分泌细胞因子与腹腔肥大细胞膜的稳定性显示文摘背景:研究表明骨髓间充质干细胞分泌的细胞因子对特异性和非特异性免疫细胞具有免疫调节作用,所以有必要深入研究其对肥大细胞的作用。目的:探讨骨髓间充质干细胞分泌因子对特异性和非特异性诱发肥大细胞脱颗粒、释放炎性递质的影响。方法:采用全骨髓贴壁法分离和纯化大鼠骨髓间充质干细胞,收集培养第3代的骨髓间充质干细胞上清液获取干细胞分泌因子。分离培养大鼠腹腔肥大细胞,分别以大鼠抗卵蛋白血清特异性被动致敏腹腔肥大细胞和用Compound48/80非特异性致敏肥大细胞。结果与结论:干细胞因子、酮替芬均能抑制特异性和非特异性诱发肥大细胞脱颗粒和释放类胰蛋白酶,与模型对照组相比差异有显著性意义(P<0.05),但干细胞因子抑制特异性和非特异性诱发肥大细胞脱颗粒和释放类胰蛋白酶的效果均不如酮替芬组(P<0.05)。结果表明干细胞分泌的细胞因子具有稳定肥大细胞膜,减轻其脱颗粒、释放炎症递质的作用。 | 高明敏 刘晓萍 于业军 贾跃伟 王家辉 李薇 | 2012 | 中国组织工程研究2012,16,27: | 2 |
| 17 | 骨髓基质干细胞分泌因子对迟发型超敏反应的影响显示文摘目的探讨骨髓基质干细胞(BMSCs)分泌因子对迟发型超敏反应的影响。方法分离培养大鼠BMSCs,贴壁法纯化,收集第三代BMSCs上清液获取干细胞分泌因子。制备小鼠接触性皮炎模型,观察干细胞分泌因子对实验小鼠耳廓肿胀的抑制作用及对胸腺指数和脾脏指数的影响。分离培养小鼠脾细胞,MTT比色法检测干细胞分泌因子对淋巴细胞增殖的影响。结果与模型组相比,干细胞分泌因子可明显减轻接触性皮炎模型小鼠耳廓肿胀程度(F=21.429,t=3.658,P<0.01),降低胸腺指数和脾指数(F=19.829、53.951,t=2.434、3.729,P<0.05),抑制ConA诱导的致敏小鼠脾淋巴细胞增殖(F=14.032,t=2.619,P<0.05)。细胞因子高剂量组对模型小鼠胸腺和脾脏淋巴细胞的抑制作用强于低剂量组(F=22.317、26.247,t=2.302、2.781,P<0.05)。结论BMSCs分泌因子对迟发型超敏反应有抑制作用。 | 王家辉 刘晓萍 于业军 高明敏 贾跃伟 李薇 | 2013 | 青岛大学医学院学报2013,49,2: | 2 |
| 18 | BMSCs及ESCs因子在迟发型超敏反应中的作用显示文摘目的探讨骨髓间充质干细胞(BMSCs)和胚胎干细胞(ESCs)因子对迟发型超敏反应(DTH)的影响。方法分离、培养并传代小鼠BMSCs和ESCs,获取BMSCs因子和ESCs因子。小鼠随机分为正常组(A组)、模型组(B组)、培养基组(c组)、地塞米松(DEX)组(D组)、BMSCs因子组(E组)和ESCs因子组(F组)6组,每组10只,应用5g/L二硝基氟苯腹部涂抹致敏、耳廓激发的方法制备小鼠变应性接触性皮炎(ACD)模型(A组不处理),A、B组尾静脉注射生理盐水,C、D、E、F组分别注射DMEM、DEX、BMSCs因子、ESCs因子,比较各组耳肿胀度、胸腺指数和脾指数,光镜下观察耳廓苏木精一伊红染色后组织学变化。结果B组耳肿胀度高于A组,差异有显著性(F=153.080,q=27.68,P<0.01);D组、E组和F组小鼠耳廓肿胀度均低于B组(q=8.130~20.440,PO.05)。镜下观察,D组、E组和F组炎性细胞浸润均较B组减少。结论BMSCs因子和ESCs因子均可抑制DTH,但ESCs因子抑制作用优于BMSCs因子。 | 吴惠惠 刘晓萍 王苗苗 于业军 秦绪艳 | 2014 | 青岛大学医学院学报2014,50,5: | 1 |
| 19 | 骨髓来源的间充质干细胞在放射性胃肠综合征治疗中的应用前景显示文摘放射性胃肠综合征(GI综合征)常见于核事故及战时的核爆炸,也是临床腹部肿瘤放疗过程中常见的并发症之一.腹盆腔受到一定剂量的辐射后,胃肠道因其黏膜上皮生长代谢活跃,对电离辐射最为敏感.间充质干细胞(MSCS)是骨髓中存在的非造血干细胞的成体干细胞,具有自我更新、高度增殖和多向分化潜能,同时具有分泌细胞因子、免疫调节及易于外源基因转染等优点,可弥补传统治疗手段的不足,将其应用于GI综合征的治疗具有广阔的应用前景.本文对MSCs在GI综合征治疗中的最新研究进展进行了综述. | 杨超 戴为民 陈海旭 吴本俨 | 2013 | 中国科学:生命科学2013,43,11: | 1 |
| 20 | 间充质干细胞与炎症性肠病的研究进展显示文摘间充质干细胞(MSC)是一种具有多分化潜能的细胞,还具有免疫调节功能。炎症性肠病(IBD)的病因和发病机制目前尚未完全明确,近年研究显示免疫紊乱在IBD的发病中起重要作用。MSC的免疫调节特性为治疗IBD提供了一种潜在的可能性。本文就MSC的免疫调节效应及其在IBD中的作用作一综述。 | 左冬梅 寿折星 范恒 刘星星 曹丹 邹舟 | 2013 | 胃肠病学2013,18,3: | 1 |