exosomes

摘要： Mild cognitive impairment(MCI)is a prodrome of Alzheimer’s disease pathology.Cognitive impairment patients often have a delayed diagnosis because there are no early symptoms or conventional diagnostic methods.Exosomes play a vital role in cell-to-cell communications and can act as promising biomarkers in diagnosing diseases.This study was designed to identify serum exosomal candidate proteins that may play roles in diagnosing MCI.Mass spectrometry coupled with tandem mass tag approach-based non-targeted proteomics was used to show the differentially expressed proteins in exosomes between MCI patients and healthy controls,and these differential proteins were validated using immunoblot and enzyme-linked immunosorbent assays.Correlation of cognitive performance with the serum exosomal protein level was determined.Nanoparticle tracking analysis suggested that there was a higher serum exosome concentration and smaller exosome diameter in individuals with MCI compared with healthy controls.We identified 69 exosomal proteins that were differentially expressed between MCI patients and healthy controls using mass spectrometry analysis.Thirty-nine exosomal proteins were upregulated in MCI patients compared with those in control patients.Exosomal fibulin-1,with an area under the curve value of 0.81,may be a biomarker for an MCI diagnosis.The exosomal protein signature from MCI patients reflected the cell adhesion molecule category.In particular,higher exosomal fibulin-1 levels correlated with lower cognitive performance.Thus,this study revealed that exosomal fibulin-1 is a promising biomarker for diagnosing MCI.

摘要： Treadmill exercise and mesenchymal stem cell transplantation are both practical and effective methods for the treatment of cerebral ischemia.However,whether there is a synergistic effect between the two remains unclear.In this study,we established rat models of ischemia/reperfusion injury by occlusion of the middle cerebral artery for 2 hours and reperfusion for 24 hours.Rat models were perfused with bone marrow mesenchymal stem cell-derived exosomes(MSC-exos)via the tail vein and underwent 14 successive days of treadmill exercise.Neurological assessment,histopathology,and immunohistochemistry results revealed decreased neuronal apoptosis and cerebral infarct volume,evident synaptic formation and axonal regeneration,and remarkably recovered neurological function in rats subjected to treadmill exercise and MSC-exos treatment.These effects were superior to those in rats subjected to treadmill exercise or MSC-exos treatment alone.Mechanistically,further investigation revealed that the activation of JNK1/c-Jun signaling pathways regulated neuronal apoptosis and synaptic-axonal remodeling.These findings suggest that treadmill exercise may exhibit a synergistic effect with MSC-exos treatment,which may be related to activation of the JNK1/c-Jun signaling pathway.This study provides novel theoretical evidence for the clinical application of treadmill exercise combined with MSC-exos treatment for ischemic cerebrovascular disease.

摘要： LncRNA(long non-coding RNA)H19 is a transcript of the H19 gene that is expressed during embryogenesis.We previously discovered a role for circular lncRNA H19 in the onset and prognosis of cerebral ischemic stroke.In this study,we used serum from patients with ischemic stroke,and mouse and cell culture models to elucidate the roles of plasma and neuronal exosomes in the regulatory effect of lncRNA H19 on insulin-like growth factor-1 and its mechanism in ischemic stroke,using western blotting,quantitative real-time polymerase chain reaction,and enzyme-linked immunosorbent assays.Plasma exosomal lncRNA H19 was negatively associated with blood levels of insulin-like growth factor-1 in samples from patients with cerebral ischemic stroke.In a mouse model,levels of exosomal lncRNA H19 were positively correlated with plasma and cerebral lncRNA H19.In a cell co-culture model,we confirmed that lncRNA H19 was transported from neurons to astrocytes by exosomes to induce downregulation of insulin-like growth factor-1 through the H19/let-7a/insulin-like growth factor-1 receptor axis.This study provides the first evidence for the transportation of lncRNA H19 by exosomes and the relationship between lncRNA H19 and insulin-like growth factor-1.

摘要： BACKGROUND:Individuals who survive a cardiac arrest often sustain cognitive impairments due to ischemia-reperfusion injury.Mesenchymal stem cell(MSC)transplantation is used to reduce tissue damage,but exosomes are more stable and highly conserved than MSCs.This study was conducted to investigate the therapeutic effects of MSC-derived exosomes(MSC-Exo)on cerebral ischemia-reperfusion injury in an in vitro model of oxygen-glucose deprivation/reperfusion(OGD/R),and to explore the underlying mechanisms.METHODS:Primary hippocampal neurons obtained from 18-day Sprague-Dawley rat embryos were subjected to OGD/R treatment,with or without MSC-Exo treatment.Exosomal integration,cell viability,mitochondrial membrane potential,and generation of reactive oxygen species(ROS)were examined.Terminal deoxynucleotidyl transferase-mediated 2’-deoxyuridine 5’-triphosphate nickend labeling(TUNEL)staining was performed to detect neuronal apoptosis.Moreover,mitochondrial function-associated gene expression,Nrf2 translocation,and expression of downstream antioxidant proteins were determined.RESULTS:MSC-Exo attenuated OGD/R-induced neuronal apoptosis and decreased ROS generation(P<0.05).The exosomes reduced OGD/R-induced Nrf2 translocation into the nucleus(2.14±0.65 vs.5.48±1.09,P<0.01)and increased the intracellular expression of antioxidative proteins,including superoxide dismutase and glutathione peroxidase(17.18±0.97 vs.14.40±0.62,and 20.65±2.23 vs.16.44±2.05,respectively;P<0.05 for both).OGD/R significantly impaired the mitochondrial membrane potential and modulated the expression of mitochondrial functionassociated genes,such as PINK,DJ1,LRRK2,Mfn-1,Mfn-2,and OPA1.The abovementioned changes were partially reversed by exosomal treatment of the hippocampal neurons.CONCLUSIONS:MSC-Exo treatment can alleviate OGD/R-induced oxidative stress and dysregulation of mitochondrial function-associated genes in hippocampal neurons.Therefore,MSCExo might be a potential therapeutic strategy to prevent OGD/R-induced neuronal injury.

摘要： The spread of SARS-CoV-2 as an emerging novel coronavirus disease (COVID-19) had progressed as a worldwide pandemic since the end of 2019. COVID-19 affects firstly lungs tissues which are known for their very slow regeneration. Afterwards, enormous cytokine stimulation occurs in the infected cells immediately after a lung infection which necessitates good management to save patients. Exosomes are extracellular vesicles of nanometric size released by reticulocytes on maturation and are known to mediate intercellular communications. The exosomal cargo serves as biomarkers in diagnosing various diseases;moreover, exosomes could be employed as nanocarriers in drug delivery systems. Exosomes look promising to combat the current pandemic since they contribute to the immune response against several viral pathogens. Many studies have proved the potential of using exosomes either as viral elements or host systems that acquire immune-stimulatory effects and could be used as a vaccine or drug delivery tool. It is essential to stop viral replication, prevent and reverse the massive storm of cytokine that worsens the infected patients’ situations for the management of COVID-19. The main benefits of exosomes could be;no cells will be introduced, no chance of mutation, lack of immunogenicity and the damaged genetic material that could negatively affect the recipient is avoided. Additionally, it was found that exosomes are static with no ability for in vivo reproduction. The current review article discusses the possibilities of using exosomes for detecting novel coronavirus and summarizes state of the art concerning the clinical trials initiated for examining the use of COVID-19 specific T cells derived exosomes and mesenchymal stem cells derived exosomes in managing COVID-19.

摘要： Nanotechnologies have been successfully applied to the treatment of various diseases.Plant-derived exosome-like nanoparticles (PENs) are expected to become effective therapeutic modalities for treating disease or in drug-delivery. PENs are minimally cytotoxic to healthy tissues, with which they show excellent biocompatibility, and are biased towards tumors by targeting specific tissues through special endocytosis mechanisms. Thus, the use of these PENs may expand the scope of drug therapies while reducing the off-target effects.In this review, we summarize the fundamental features and bioactivities of PENs extracted from the grape, grapefruit, ginger, lemon, and broccoli and discuss the applications of these particles as therapeutics and nanocarriers.

摘要： Blood exosomes,which are extracellular vesicles secreted by living cells into the circulating blood,are regarded as a relatively noninvasive novel tool for monitoring brain physiology and disease states.An increasing number of blood cargo-loaded exosomes are emerging as potential biomarkers for preclinical and clinical Alzheimer's disease.Therefo re,we conducted a meta-analysis and systematic review of molecular biomarkers derived from blood exosomes to comprehensively analyze their diagnostic performance in preclinical Alzheimer's disease,mild cognitive impairment,and Alzheimer's disease.We performed a literature search in PubMed,Web of Science,Embase,and Cochrane Library from their inception to August 15,2020.The research subjects mainly included Alzheimer's disease,mild cognitive impairment,and preclinical Alzheimer's disease.We identified 34 observational studies,of which 15 were included in the quantitative analysis(Newcastle-Ottawa Scale score 5.87 points)and 19 were used in the qualitative analysis.The meta-analysis results showed that core biomarkers including Aβ_(1-42),P-T181-tau,P-S396-tau,and T-tau were increased in blood neuro nderived exosomes of preclinical Alzheimer's disease,mild cognitive impairment,and Alzheimer's disease patients.M olecules related to additional risk facto rs that are involved in neuroinflammation(C1q),metabolism disorder(P-S312-IRS-1),neurotrophic deficiency(HGF),vascular injury(VEGF-D),and autophagy-lysosomal system dysfunction(cathepsin D)were also increased.At the gene level,the differential expression of transc ription-related factors(REST)and microRNAs(miR-132)also affects RNA splicing,transport,and translation.These pathological changes contribute to neural loss and synaptic dysfunction.The data confirm that the above-mentioned core molecules and additional ris k-related factors in blood exosomes can serve as candidate biomarkers for preclinical and clinical Alzheimer's disease.These findings support further development of exosome biomarkers for a clinical blood test for Alzheimer's disease.This meta-analysis was registered at the International Prospective Register of Systematic Reviews(Registration No.CRD4200173498,28/04/2020).

摘要： After organ and tissue injury,the proliferation and differentiation of stem cells themselves play only a small role in the repair of injury,and their repair role is mainly played through the paracrine function of stem cells.Exosomes are nano-scale vesicles that are secreted into the extracellular space in an exocytic manner,and its own function will be regulated after the target cells absorb the exosomes.Stem cell-derived exosomes communicate between cells by transmitting proteins,lipids and micro-RNAs(miRNAs).The targeting and biological properties of stem cell-derived exosomes are determined by the level of miRNAs that they carry.After the exosomes reach the target cells and undergo fusion,the gene expression of the target cells is changed by degradation and expression.In addition,the RNA and protein of stem cell-derived exosomes can also limit the development of injury through cell homing.This article will review the mechanism of stem cell-derived exosomes in wound healing,joint injury,fracture healing and cardiac injury.

摘要： Poor healing of cutaneous wounds is a common medical problem in the field of traumatology.Due to the intricate pathophysiological processes of wound healing,the use of conventional treatment methods,such as chemical molecule drugs and traditional dressings,have been unable to achieve satisfactory outcomes.Within recent years,explicit evidence suggests that mesenchymal stem cells(MSCs)have great therapeutic potentials on skin wound healing and regeneration.However,the direct application of MSCs still faces many challenges and difficulties.Intriguingly,exosomes as cell-secreted granular vesicles with a lipid bilayer membrane structure and containing specific components from the source cells may emerge to be excellent substitutes for MSCs.Exosomes derived from MSCs(MSC-exosomes)have been demonstrated to be beneficial for cutaneous wound healing and accelerate the process through a variety of mechanisms.These mechanisms include alleviating inflammation,promoting vascularization,and promoting proliferation and migration of epithelial cells and fibroblasts.Therefore,the application of MSC-exosomes may be a promising alternative to cell therapy in the treatment of cutaneous wounds and could promote wound healing through multiple mechanisms simultaneously.This review will provide an overview of the role and the mechanisms of MSC-derived exosomes in cutaneous wound healing,and elaborate the potentials and future perspectives of MSC-exosomes application in clinical practice.

摘要： BACKGROUND Liver cancer is the fourth most significant cause of cancer-related death.Lack of early diagnosis strategy and a scarcity of efficient therapy constitute the main reasons for its lethality.Exosomes,which contain various bioactive molecules,are characterized by high biocompatibility,low immunogenicity,and high transport efficiency.As a result,exosomes have become a research hotspot and present significant potential for cancer diagnosis biomarkers,biotherapeutics,therapy targets,drug carriers and therapeutic agents.AIM To explore the potential of exosomes in the diagnosis and treatment of liver cancer.METHODS We conducted a systematic literature search via PubMed and Web of Science.The following keywords were used:"exosomal biomarkers","exosomal therapy","exosomal therapy",and"liver cancer"or"HCC".The duplicate data were deleted by EndNote software.Literature search focused on full-texts and references of each article were carefully checked.One author(Xiao-Cui Wei)screened the literature that met the following inclusion criteria:(1)Detection of exosomes or their contents in clinical samples(body fluid or tissue);or(2)Exosomes served as drug carriers or therapeutic factors.Two authors(Xiao-Cui Wei and Li-Juan Liu)independently reviewed all retained literature and analyzed the information.RESULTS A total of 1295 studies were identified using the systematic literature search.Of these,835 duplicate studies were removed.A further 402 irrelevant studies were excluded due to being irrelevant,including other diseases,review articles,the literature containing neither clinical samples nor animal experiments,exosomeindependent studies,methods for detecting exosomes,or articles in Chinese.Finally,58 published papers were retained and analyzed in the study.It showed a list of potential exosomal biomarkers that were upregulated in the blood samples of patients with liver cancer.Those downregulated in exosomes might serve as possible biotherapeutics.Some exosomes derived from cells in vitro were used for cytology or animal experiments to explore the mechanism of these exosome contents in disease.These contents might serve as potential targets for liver cancer.Additionally,we also discussed that exosomes serve as drug carriers or therapeutic factors.CONCLUSION Exosomes might serve as potential biomarkers or therapeutic biotargets in liver cancer and have the potential to act as drug carriers and self-treatment factors for liver cancer patients.

摘要： 本发明涉及一种离散汲取收集外泌体的方法及试剂，所述试剂包括PBS缓冲液、固定液及原水，所述方法包括：首先用一种抗凝溶剂将外泌体与DNA/RNA的大分子团分离，然后分级离心，除去DNA/RNA等大分子团，留存纯度较高的外泌体。其优点在于：(1)本发明提取外泌体的试剂可高效提取外泌体，收率高、纯度高。(2)本发明操作简单，成本低，对实验操作人员技术、实验设备要求低，有很好的应用前景，拓展了外泌体研究领域，对于外泌体的研究有重大意义。

摘要： 本发明提供了一种携带抗肿瘤蛋白靶向性exosome的制备方法，包括以下步骤：(1)构建表达肿瘤靶向性多肽iRGD、exosome膜蛋白lamp2b及抗肿瘤蛋白PINT‑87aa的融合基因表达质粒；(2)将表达质粒转染到人的胚肾细胞HEK293细胞中，puro抗生素筛选获得稳定表达抗肿瘤融合蛋白的细胞系；(3)细胞扩增培养，对细胞释放出来的携带抗肿瘤蛋白的exosome进行分离纯化。本发明的方法成功构建了一种仿生物源性抗肿瘤蛋白输送载体，能够高效抑制肿瘤的生长，为恶性的治疗提供了一种潜在的理想的治疗方法。

摘要： 本发明涉及人脐带间质干细胞分泌的exosome治疗皮肤损伤的作用，具体涉及一种脐带间质干细胞分泌的直径为100nm左右的囊泡结构exosome在促进皮肤损伤修复中的药用作用，属于皮肤创伤治疗的技术领域；本发明所述的exosome是来源于人脐带间质干细胞，经试验验证，所涉及的exosome作为脐带间质干细胞细胞分泌的重要“成分”对皮肤损伤方面（包括烧烫伤和切除损伤）具有明显促修复的药用作用，而且其具有便于储存和运输的优势，这样脐带间质干细胞来源的exosome就为以间质干细胞为基础的非细胞治疗皮肤损伤提供了可能。

摘要： 本发明涉及一种缓释iPSC‑MSC‑Exosomes温敏壳聚糖复合凝胶制备方法。技术方案是：缓释iPSC‑MSC‑Exosomes温敏壳聚糖复合凝胶制备方法，按以下步骤进行：1)将壳聚糖盐酸盐和明胶粉末溶于水中搅拌混匀；2)在步骤1)获得的溶液中逐滴加入0.444g/ml甘油磷酸二钠盐溶液，使得溶液中的壳聚糖：明胶：甘油磷酸二钠盐的最终比等于5:1.5‑2.5:7‑13，得到温敏壳聚糖凝胶溶液；3)在体外通过超速离心法提取iPSC‑MSC‑Exosomes；4)将iPSC‑MSC‑Exosomes逐渐加入温敏壳聚糖溶液中混合，得到温敏壳聚糖复合凝胶。该方法可持续释放外泌体以促进组织损伤修复。

摘要： 本发明公开了一种尿液中近端肾小管来源exosome含量的检测方法，包括以下步骤：S1，制备尿液exosome悬液；S2，测定尿液exosome悬液中蛋白浓度；S3，分选尿液exosome悬液中近端肾小管来源exosome；S4，对S3中分选出的尿液中近端肾小管来源的exosome进行检测。本发明采用超速离心方法获得高纯度的尿液exosome，再采用免疫磁珠吸附exosome，克服了exosome体积小的问题，增加了流式细胞仪方法检测exosome的可行性；采用CD63和CD13免疫标记方法分选尿液中近端小管来源的exosome，并用于后续对尿液中近端肾小管来源exosome含量进行准确检测分析。

摘要： 本发明提供了一种肿瘤源性的exosome来源的与人类肝癌相关的miRNA标志物及其在诊断人类肝癌中的用途，所述miRNA该标志物为exosome来源的miRNA‑46a。本发明还提供了检测exosome来源的miRNA‑46a的试剂、含有该检测试剂的试剂盒及其在诊断人类肝癌中的用途。本发明还提供了用于检测exosome来源的miRNA‑46a的方法及其在诊断人类肝癌中的用途。本发明还提供了从少量血清中快速提取exosome的试剂、含有该exosome提取试剂的试剂盒以及使用该试剂盒从血液中提取exosome的方法。本发明的miRNA标志物及其检测试剂可用于肝癌的早期诊断，其灵敏性和特异性均优于现有技术。

摘要： 本发明提供了一种肿瘤源性的exosome来源的与人类肝癌相关的miRNA标志物及其在诊断人类肝癌中的用途，所述miRNA该标志物为exosome来源的miRNA‑33b。本发明还提供了检测exosome来源的miRNA‑33b的试剂、含有该检测试剂的试剂盒及其在诊断人类肝癌中的用途。本发明还提供了用于检测exosome来源的miRNA‑33b的方法及其在诊断人类肝癌中的用途。本发明还提供了从少量血清中快速提取exosome的试剂、含有该exosome提取试剂的试剂盒以及使用该试剂盒从血液中提取exosome的方法。本发明的miRNA标志物及其检测试剂可用于肝癌的早期诊断，其灵敏性和特异性均优于现有技术。

摘要： 本实用新型公开了一种用于体液样品exosome保存的试剂盒，包括盒体，所述盒体顶部活动连接有顶盖，所述顶盖与盒体之间设置有铰链，所述顶盖顶部外表面固定安装有把手，所述顶盖前壁固定安装有子扣，所述盒体前壁与子扣相对应位置固定安装有母扣，所述盒体内部设置有安装板，所述安装板内部设置有弹簧，所述弹簧一侧固定连接有第一短杆，所述第一短杆一侧末端固定连接有第一弧形夹板，所述安装板内壁中心轴线上固定连接有竖杆，本实用新型通过设置隔间、导热板及保温板，将冰块存放在隔间内部，避免打湿试剂盒底部，且导温板可以使得安装板内部温度保持在适合的温度左右，保温板也可以防止冰块过早的融化，该装置结构简单，方便实用。

摘要： 本发明提供了一种exosomes装载的miR‑93‑5p在治疗类风湿性关节炎中的应用，属于生物医学技术领域；本发明首先制备过表达或阻断miR‑93‑5p的G‑exosomes，并通过体外实验证明，过表达miR‑93‑5p的G‑exosomes更加明显的抑制了Th17细胞的分化；并通过研究过表达和阻断miR‑93‑5p的G‑exosomes对于CIA模型小鼠发病的作用，发现过表达miR‑93‑5p的G‑exosomes对CIA小鼠的保护作用更强了；而阻断miR‑93‑5p的G‑exosomes则没有上述作用；结果表明过表达miR‑93‑5p的G‑exosomes可用于类风湿性关节炎的治疗。

摘要： 本发明涉及一种NK细胞exosomes可控规模化的制备方法，包括以下步骤：(1)收集NK细胞培养基上清液，并用磷酸盐缓冲液进行稀释，得到稀释液；(2)将稀释液依次在低转速和高转速下离心，且每次离心后取上清液弃沉淀；(3)将离心上清液置于超滤管中进行离心，取上清液，得到的浓缩上清液进行超高速离心，超高速离心后取沉淀；(4)将步骤(3)超高速离心后得到的沉淀置于磷酸盐缓冲液中并转移至透析袋中进行透析，即得所述NK细胞exosomes。与现有技术相比，本发明粒径分布均匀，且表面携带标志性蛋白CD9、CD81、CD63，克服了现有制备技术存在纯度低，产量低，表征蛋白表达量低等不足，且可以直接用于后续的检测，安全系数高。