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??Objective    To isolate and identify exosomes derived from stem cells from apical papilla. Methods??Stem cells from apical papilla ??SCAP?? were primarily cultured and identified. SCAP-derived exosomes were isolated by ultracentrifugation. The morphology was observed by transmission electron microscopy. The markers CD9 and Alix were detected by Western blotting. Results??SCAP could release exosomes??which showed a cup-like microvesicle structure??with a diameter of about 30-100 nm under electronic microscopy. The specific markers CD9 and Alix were positively expressed. Conclusion??The results show that exosomes can be successfully isolated from culture supernatants of SCAP by ultracentrifugation.     

Urinary biomarkers of prostate cancer

The development of more specific biomarkers for prostate cancer and/or high‐risk prostate cancer is necessary, because the prostate‐specific antigen test lacks specificity for the detection of prostate cancer and can lead to unnecessary prostate biopsies. Urine is a promising source for the development of new biomarkers of prostate cancer. Biomarkers derived from prostate cancer cells are released into prostatic fluids and then into urine. Urine after manipulation of the prostate is enriched with prostate cancer biomarkers, which include prostate cancer cells, DNAs, RNAs, proteins and other small molecules. The urinary prostate cancer antigen 3 test is the first Food and Drug Administration‐approved RNA‐based urinary marker, and it helps in the detection of prostate cancer on repeat biopsy. The SelectMDx test is based on messenger RNA detection of DLX1 and HOXC6 in urine after prostate massage, and helps in the detection of high‐risk prostate cancer on prostate biopsy. Exosomes are extracellular vesicles with a diameter of 30–200 nm that are secreted from various types of cells. Urinary prostate cancer‐derived exosomes also contain RNAs and proteins specific for prostate cancer (e.g. PCA3 and TMPRSS2‐ERG), and could be promising sources of novel biomarker discovery. The ExoDx Prostate test is a commercially available test based on the detection of three genes (PCA3, ERG and SPDEF) in urinary exosomes. Advancement of comprehensive analysis (microarray, mass spectrometry and next‐generation sequencing) has resulted in the discovery of several urinary biomarkers. Non‐invasive urinary markers can help in the decision to carry out prostate biopsy or in the design of a therapeutic strategy.     

胃癌患者血清外泌体中DANCR表达及其临床意义

目的检测胃癌患者血清外泌体(exosome)中抗分化非编码RNA(DANCR)的表达水平并分析其临床应用价值。方法采用实时荧光定量RT-PCR检测胃癌细胞培养上清和胃癌患者血清exosome中DANCR表达水平,分析其与临床病理资料的相关性,绘制ROC曲线评价其诊断效能。结果 DANCR在人胃癌细胞系HGC-27中表达水平(4.43±0.37)明显高于胃黏膜上皮细胞(1.01±0.01),差异有统计学意义(P0.05)。HGC-27细胞培养上清exosome中DANCR表达水平(3.06±0.14)明显高于GES-1细胞(1.01±0.20),差异有统计学意义(P0.05)。DANCR在胃癌患者血清exosome中的平均含量[5.060(1.380,22.785)]较胃良性疾病患者[0.535(0.340,1.380)]和体检健康者[1.200(0.605,1.655)]明显升高(Z分别为-3.409,-4.229,P均0.01),且与肿瘤大小、TNM分期以及淋巴结转移相关。胃良性疾病患者与体检健康者相比,血清exosome中DANCR含量差异无统计学意义(Z=-1.308,P0.05)。胃癌患者血清exosome中DANCR的ROC曲线下面积(AUCROC)为0.777,95%可信区间(CI)为0.678~0.876,cut-off值2.50,敏感性为68.6%,特异性为84.4%。结论胃癌患者血清exosome中DANCR表达水平升高,有望成为胃癌诊断的新指标。     

肿瘤来源的外体及其在肿瘤中的作用

外体（exosome）是由细胞内多泡体（multivesicular body,MVB）与细胞膜融合而释放到细胞外的纳米大小的膜性小囊泡,具有一定的免疫调节功能。造血细胞以及非造血细胞都可以分泌外体。肿瘤来源的外体（tumor-derived exosome,TEX）存在于肿瘤细胞培养上清液、肿瘤患者血浆或者恶性渗出液中,包含天然的肿瘤相关抗原,并能将其呈递给T细胞,可有效地促进细胞毒T淋巴细胞（cytotoxic T lymphocyte ,CTL）的活化,产生抗瘤免疫。近年来,也有文献报道外体可诱导免疫耐受,甚至产生免疫抑制作用而使得肿瘤逃逸宿主免疫系统对它的免疫监视,并能促进肿瘤细胞的增殖。这些功能的不一致与外体的表型密切相关。     

基于网络药理学及全转录组测序对当归四逆汤干预外泌体抑制多发性骨髓瘤血管新生的机制研究

目的 基于骨髓瘤细胞株外泌体测序及网络药理学研究当归四逆汤抑制多发性骨髓瘤血管新生的机制,为后续研究提供有针对性的指导.方法 利用TCMSP在线平台及SwissTargetPrediction网站获取当归四逆汤主要化学成分及对应靶点,借助GeneCards获取多发性骨髓瘤血管新生相关靶点基因,经与当归四逆汤作用靶点匹配...     

视网膜损伤：外泌体转录组学的研究进展

衰老、缺氧、高糖刺激等多种因素导致的视网膜损伤过程中,外泌体发挥着重要作用。外泌体是一种纳米级的细胞外囊泡,包含微小RNA、环状RNA等多种转录产物,广泛参与视网膜色素上皮细胞损伤、新生血管形成、视网膜神经节细胞和光感受器细胞凋亡等过程。通过转录组学分析外泌体对视网膜损伤及修复的调控机制,识别糖尿病视网膜病变、青光眼等眼科疾病相关分子生物标志物,寻找新的治疗靶点,已成为视网膜研究的热点。因此,本文对外泌体转录组学在视网膜损伤中的研究进展进行综述。     

急性期川崎病患儿血清外泌体蛋白质组学的前瞻性研究

目的 筛选急性期川崎病(Kawasaki disease,KD)患儿血清外泌体蛋白,分析其生物学过程及功能,以期为早期临床诊断KD提供新的生物标志物.方法 前瞻性选取2019年6月至2020年8月于苏州大学附属儿童医院治疗的KD患儿(n=13)纳入KD组;同期选取因细菌感染入该院治疗的患儿(n=13)纳入感染对照组.采...     

血清外泌体miR-451a在弥漫大B细胞淋巴瘤治疗监测中的意义

目的 探索血清外泌体miR-451a在弥漫大B细胞淋巴瘤（diffuse large B cell lymphoma, DLBCL）中的水平及其在治疗监测中的价值。方法 本研究共纳入56例DLBCL患者，56例健康对照者。收集新发DLBCL患者治疗前、化疗2~4疗程及化疗结束后血清样本，并同时收集健康人血液标本， 提取血清中的外泌体RNA，并进行实时荧光定量PCR（quantitative real time polymerase chain reaction, qRT-PCR），用受试者工作特征（receive operator characteristic, ROC）曲线判定miR-451a的诊断效能，用各时点收集的血清样本动态分析血清外泌体miR-451a水平与化疗效果之间的关系。结果 56例DLBCL患者与56例健康对照者相比，DLBCL患者血清外泌体miR-451a水平下降（P<0.000 1），在两组受试者间用miR-451a诊断DLBCL的曲线下面积(AUC)为0.737（95%CI0.645~0.816）。在随访到的43例DLBCL患者中，化疗后获得完全缓解或者部分缓解的患者其血清外泌体miR-451a水平较化疗前有所上升（P<0.05），与配对的健康人水平差异无统计学意义（P>0.05）；化疗后未获得缓解的患者，其血清外泌体miR-451a水平较化疗前无明显变化（P>0.05），且仍然低于配对的健康人水平（P<0.05）；化疗完成后在未缓解者与缓解者之间进行鉴别，血清miR-451a的AUC为0.867（95%CI0.728~0.951）。结论 血清外泌体miR-451a水平动态监测有助于DLBCL化疗过程中的疗效（是否缓解）判断。