Stem cells to reverse aging

As human life expectancy continues to increase and the birth rate continues to decline, the phenomenon of aging is becoming more prominent worldwide. Therefore, addressing the problems associated with global aging has become a current research focus. The main manifestations of human aging are structural degeneration and functional decline of aging tissues and organs, quality of life decline, decreased ability to resist diseases, and high incidence rates of a variety of senile degenerative diseas...     

奥沙利铂联合Exosome诱导的效应性T细胞抑制HepG2细胞增殖的研究

目的观察奥沙利铂(OXA)联合负载HepG2细胞裂解物的Exosome诱导的效应性T细胞对肝癌HepG2细胞系增殖的抑制作用。方法自健康人外周血中提取单个核细胞和T淋巴细胞,并以GM-CSF+IL-4的培养方案获得树突状细胞(DC)。将反复冻融后产生的HepG2细胞裂解物体外致敏DC,收集培养上清后以超高速离心法分离得到Exosome,电镜下观察其形态特征。用致敏DC及其分泌的Exosome刺激T淋巴细胞的增殖和活化。以ELISA试验检测不同刺激环境下T细胞分泌IL-1β、IL-2、IL-4、IL-6、IL-10、IFN-γ和TNF-α的能力。以细胞杀伤实验(MTT法)检测不同因素刺激后的T细胞、OXA(2.5、5.0、10.0mg.L-1)以及两者联合应用对HepG2细胞生长的抑制效应。结果与未致敏DC及其分泌的Exosome相比,HepG2细胞冻融裂解物致敏的DC及其分泌的Exosome能显著促进T细胞的增殖,并使其活化和分泌大量的IL-1β、IL-2、IL-6、IFN-γ和TFN-α等Th1型细胞因子。活化后的T细胞、OXA以及两者联合应用对HepG2细胞体外增殖均有抑制作用,而活化T细胞联合10mg.L-1OXA对HepG2细胞的杀伤率最高,但与联合5 mg.L-1OXA的效应相比,差异无统计学意义(P0.05)。结论负载肿瘤细胞抗原的DC所分泌的Exosome可在体外刺激T细胞,使其活化为具有抗肿瘤效应的细胞毒性T细胞,该细胞联合低浓度OXA能显著抑制肝癌细胞的体外增殖,并降低OXA的用量。     

Significance of trogocytosis and exosome-mediated transport in establishing and maintaining the tumor microenvironment in lymphoid malignancies

It is widely accepted that the tumor microenvironment plays an important role in the progression of lymphoid malignancies. Interaction between the tumor and its surrounding immune cells is considered a potential therapeutic target. For example, anti-programmed cell death 1 (PD-1) antibody stimulates the surrounding exhausted immune cells to release PD-1/PD-L1, thereby leading to the regression of PD-L1-positive tumors. Recently, biological phenomena, such as trogocytosis and exosome-mediated transport were demonstrated to be involved in establishing and maintaining the tumor microenvironment. We found that trogocytosis-mediated PD-L1/L2 transfer from tumor cells to monocytes/macrophages is involved in immune dysfunction in classic Hodgkin lymphoma. Exosomes derived from Epstein-Barr virus (EBV)-associated lymphoma cells induce lymphoma tumorigenesis by transferring the EBV-coding microRNAs from the infected cells to macrophages. In this review, we summarized these biological phenomena based on our findings.     

外泌体miRNA在肺癌诊疗中的研究进展

外泌体是一种几乎所有细胞都能分泌的胞外小泡,在细胞和器官之间运输脂质、蛋白质和核酸。作为一种天然细胞间通信器,外泌体广泛分布在生物体液中。外泌体中的微小RNA(miRNA)在肿瘤的发生、发展和侵袭转移中起至关重要的作用,并具有成为肿瘤生物标志物的潜力。本文综述了外泌体中miRNA在肺癌诊断、预后评估及药物疗效预测中的作...     

肺移植免疫学相关基础与临床研究进展

器官移植受者的生存质量与免疫状态密切相关。与其他实体器官移植相比,肺移植受者的长期预后较差,其涉及的免疫学机制更为复杂,既有共性又有特性。因此,深入了解同种异体肺移植免疫应答过程中的免疫学机制对于改善肺移植受者的长期生存尤为重要。本文从肺脏免疫细胞组成的独特性、肺移植术后排斥反应特点、肺移植病原体感染的早期预警和鉴别诊断以及肺移植相关并发症等方面进行探讨,总结同种异体肺移植涉及免疫学相关的临床诊断和基础研究进展,旨在阐述肺移植的免疫学特点,为临床提高肺移植受者的长期生存率以及移植物失功的防治水平等提供理论依据。     

基于唾液转录组学探讨慢性胃炎和胃癌脾虚证的分子机制

中医证候研究是中医药学的研究热点,基于唾液转录组学探讨疾病证候的复杂性并揭示其科学内涵具有巨大科研潜力。现分析了慢性胃炎和胃癌脾虚证的联系以及疾病的进展过程。基于唾液外泌体exoRNA表达谱,运用唾液转录组学的学科优势,在中医学同病异证,异病同证的理论指导下,提出寻找其具有临床价值的差异表达的分子标志物。进一步推动证候物质基础的定量、定性研究,并从分子机制角度切入促进中医肿瘤证候的精准化发展。     

环磷酰胺致小鼠心脏功能损伤超声评价及其与血清外泌体miRNA的相关性

目的分析环磷酰胺心脏毒性小鼠模型血清外泌体miRNA的改变,为更准确的评估环磷酰胺导致心脏功能损伤的程度提供新方法。 方法选用7~8周龄的C57BL/6小鼠,随机分为对照组和环磷酰胺组2组。环磷酰胺组小鼠给予环磷酰胺100 mg /（kg?周）,根据临床化疗方案,连续腹腔注射2周后,休息1周,再连续注射2周,共5周;对照组小鼠腹腔注射等量0.9%氯化钠溶液。第5周进行小鼠心脏超声检查,测量或计算左心室射血分数（LVEF）、短轴缩短率（FS）、左心室等容舒张期时间（IVRT）及Tei指数,同时行血清外泌体候选miRNA的q-PCR检测。采用t检验比较环磷酰胺组与对照组小鼠LVEF、FS、IVRT、Tei指数、血清外泌体miRNA表达量。 结果与对照组小鼠比较,环磷酰胺组小鼠LVEF、FS均降低［0.62±0.05 vs 0.72±0.06,（28.00±3.10）% vs （35.10±4.95）%］,且差异均有统计学意义（t=3.591,P=0.003;t=3.453,P=0.004）;Tei指数、IVRT均升高［0.71±0.17 vs 0.59±0.14,（27.42±8.42）ms vs （23.40±6.70）ms］,但差异无统计学意义。与对照组小鼠比较,环磷酰胺组小鼠血清外泌体中miR-133a-3p、miR-146a-5p表达增高（1.931±1.460 vs 0.072±0.077,1.895±1.059 vs 0.790±0.296）,且差异均有统计学意义（t=4.072,P=0.001;t=3.168,P=0.006）;miR-30c-5p、miR-99a-5p表达差异无统计学意义（0.870±0.327 vs 0.530±0.670,0.404±0.134 vs 0.714±0.404）。 结论C57BL/6小鼠腹腔注射环磷酰胺能够引起心脏功能明显损伤,心脏功能受损小鼠血清外泌体中miR-133a-3p和miR-146a-5p表达增高,使其有望成为诊断环磷酰胺心脏功能受损新的血清标志物。     

大鼠骨髓间充质干细胞exosome提取及其心肌细胞H9C2靶向作用的实验探索

目的 研究缺氧预处理条件下,骨髓间充质干细胞分泌的exosome的特征;探索心肌细胞H9C2是否骨髓间充质干细胞exosome作用的靶细胞之一,以及H9C2对骨髓间充质干细胞exosome的摄取是否随时间有一定的变化特征。方法 采用分步离心结合蔗糖/D2O垫超速离心方法分离提取干细胞分泌的exosome囊体颗粒。采用透射电镜和Western blotting法鉴定提取物是否为exosome。采用图像分析软件Image- Pro Plus 6.0 对透射电镜结果中exosome 数据进行采集,采用excel和统计软件Graph Pad 5.0对采集得到的exosome数据进行统计,以明确exosome的直径分布区间和平均半径等特征。采用绿色荧光染料PKH-67标记exosome,将标记的exosome与H9C2共孵育,观察exosome能否被心肌细胞H9C2摄取。将exosome与H9C2共孵育的不同时间段,观察H9C2对exosome的摄取随时间变化的特征。结果 提取物呈微型囊体结构,形态近似球形或者椭球形,大小均一,均匀分散的分布在视野中,提取物均阳性表达CD63和CD9分子,且较CD63分子,提取物更高表达CD9分子;缺氧预处理条件下,骨髓间充质干细胞exosome直径的集中分布范围是20～60nm, 半径为(17.03 ± 0.40) nm;exosome与心肌细胞H9C2共孵育结果显示,仅实验组H9C2细胞质内可见大量的exosome绿色荧光颗粒;H9C2摄取exosome的时间特征显示,对照组4个亚组中H9C2对exosomes的摄取情况无明显不同。实验组各亚组显示,exosome与H9C2共孵育1h,H9C2细胞质中即开始观察到标记的exosome;共孵育1.5～2.5h,H9C2细胞质中exosome绿色荧光颗粒数量较共孵育其他时间段的多,荧光强度较共孵育其他时间段的强。结论 成功分离提取到了缺氧预处理条件下的大鼠骨髓间充质干细胞exosome;心肌细胞H9C2是骨髓间充质干细胞exosome生物学作用的靶细胞;H9C2对骨髓间充质干细胞exosome的摄取情况随时间的延长有明显的变化。     

Conceptus implantation and placentation: molecules related to epithelial–mesenchymal transition,lymphocyte homing,endogenous retroviruses,and exosomes

Processes of conceptus implantation and placentation, unique to mammalian reproduction, have been extensively studied. It was once thought that processes of these events varied greatly, notably between invasive and noninvasive modes of implantation and/or placentation. Regardless of the mode of implantation, however, physiological and biochemical processes in conceptus implantation to the maternal endometrium including the kinds of gene expression and their products are now considered not to differ so much. Recent progress has identified that in addition to the hormones, cytokines, proteases and cell adhesion molecules classically characterized, epithelial–mesenchymal transition, molecules related to lymphocyte homing, the expression of endogenous retroviruses and possibly exosomes are all required for the progression of conceptus implantation to placentation. In this review, therefore, new findings related to these events are integrated into the context of conceptus implantation to the maternal endometrium.     

Exosomal miRNA derived from keratinocytes regulates pigmentation in melanocytes

Background

Pigmentation is controlled by complex mechanisms. Evidence suggests that miRNAs can regulate pigmentation. However, the mechanism has not been fully elucidated. Objective In this study, we revealed a novel mechanism that regulates pigmentation involving exosomes, miRNAs and the crosstalk between keratinocytes and melanocytes.