单细胞测序技术在胃癌研究中的应用

胃癌是一种高异质性肿瘤,传统测序方式通常将胃癌组织或细胞当做整体进行分析,易忽视其中各个细胞亚群的作用。作为一种新兴技术,单细胞测序可在单个细胞水平对基因组、转录组等多组学进行分析,可识别胃癌组织中的罕见细胞亚群,有助于解析胃癌的发生发展轨迹。本文就单细胞测序的优势、流程以及在胃癌研究中的应用进行综述,并探讨其应用前景...     

单细胞测序技术在胰腺癌领域应用现状和展望

胰腺癌是预后极差的恶性肿瘤,其特征是癌细胞与肿瘤微环境中存在复杂的相互作用网络和极高的异质性,导致缺乏早期诊断和有效治疗的策略.新兴的单细胞测序技术从单细胞层面进行基因组、转录组或表观遗传学分析,揭示疾病内在的分子机制,相比于传统的研究方法,更有助于阐明肿瘤内异质性问题.目前,单细胞测序技术已广泛应用于胰腺癌领域的研究...     

单细胞转录组测序技术在骨关节炎中的研究进展

骨关节炎是一种常见的以软骨细胞退变、软骨下骨重构以及滑膜炎症为主要特征的慢性退行性骨关节疾病,目前病因及分子发病机制尚不明确.单细胞转录组测序可以对全转录组在单细胞水平进行扩增与测序,可以解决普通细胞测序技术不能精确到细胞类型的问题,用于细胞异质性发现及微量样品的转录组分析,可以在分子水平上研究骨关节炎的发病机制和病理...     

单细胞测序技术在膀胱癌研究中的应用

单细胞测序技术是在单细胞水平上, 对基因组、转录组、表观组进行高通量测序分析的一项新技术。目前, 单细胞测序技术已广泛应用于膀胱癌的研究, 通过识别不同的细胞亚群、免疫微环境, 开辟了解膀胱细微肿瘤生物学的新途径, 并且单细胞测序技术有望通过识别和使用新的生物标志物和靶向治疗, 对膀胱癌的诊断和治疗做出重要改变。本文总结单细胞测序技术在膀胱癌的异质性细胞亚群、发生发展规律、免疫微环境和耐药性等方面中最新的研究应用进展。     

单细胞测序技术在膀胱癌研究中的应用进展

膀胱癌是泌尿系统最常见的恶性肿瘤之一, 其复杂的肿瘤微环境和极高的肿瘤内异质性, 导致膀胱癌早期诊断困难及治疗效果欠佳。新兴的单细胞测序技术是一项对单细胞进行基因组、转录组、代谢组及表观遗传学测序, 进而识别细胞间精细差异的强大技术, 对于揭示肿瘤内异质性及探讨肿瘤微环境中细胞间的复杂关系具有独特优势。本文中, 我们总结了单细胞测序技术在膀胱癌肿瘤内异质性、肿瘤微环境、治疗耐药性及治疗靶点等方面的研究, 以期为膀胱癌的诊疗及研究提供新的思路。     

单细胞RNA测序在肾脏疾病研究中的应用与前景

科学家们通过单细胞RNA测序(single-cell RNA sequencing,scRNA-seq)技术对疾病发生、发展过程中不同细胞所发挥的独特作用认识越来越深刻.scRNA-seq方法也正逐渐应用于肾脏疾病研究领域.本文将介绍scRNA-seq技术的发展,并总结scRNA-seq技术在肾脏领域的正常细胞图谱的绘...     

单细胞测序技术在骨关节疾病方面的应用

在关节疾病的发病机制的研究上,目前最常针对的对象还是骨、软骨及滑膜组织,以及从这些组织的整个病理改变的方向去探索可能的发病过程及发病机制,所以存在很多局限性和不准确性,比如忽略了疾病组织内部的单个细胞的异质性在疾病当中所发挥的作用。单细胞测序技术(single-cell sequencing)是在单个细胞水平上对基因组、转录组及表观基因组进行测序的技术,能够从组织样本中获得不同细胞间的异质性信息和对珍贵微量样本进行测序,从而更深层次地反映生命规律与本质,本文就单细胞测序技术的基本方法及在关节疾病方面的应用进行概述。     

单细胞转录组测序在肿瘤浸润免疫细胞中的研究进展

免疫细胞与肿瘤进展及治疗密切相关。单细胞转录组测序(single-cell RNA sequencing, scRNA-seq)已广泛应用于解析肿瘤浸润免疫细胞的生物学行为、机制及其与机体的关系。scRNA-seq能全面剖析肿瘤浸润免疫细胞并促进临床个体化精准治疗。     

单细胞RNA测序在肾脏领域的研究进展

肾脏是结构和功能高度复杂的器官,由多种细胞类型组成,明确各种类型及其亚型细胞在生理和病理状态下基因转录图谱及其变化对阐明肾脏结构、功能以及疾病的发病机制极为重要。单细胞RNA测序(single-cell RNA sequencing,scRNA-seq)的发展引起转录组学研究模式的转变,从对大量组织基因转录的平均水平的分析转向对特定器官(或组织)中单细胞基因转录的细胞水平的研究。本文在阐述scRNA-seq技术及其揭示的新发现的肾脏细胞亚型和功能的基础上,重点对常见的多种肾脏疾病肾组织中免疫细胞scRNA-seq的研究进展进行综述。     

单细胞测序技术及其在结直肠癌中的研究进展

结肠直肠癌(CRC)是全球第三常见及第二位肿瘤致死癌症疾病, 由于基因突变的发生及肿瘤转移等事件造成CRC的预后不良。单细胞测序技术可以在单细胞水平上进行基因组、转录组、表观遗传学的高通量测序分析, 在肿瘤免疫微环境分析、肿瘤异质性检测、肿瘤转移机制探究及循环肿瘤细胞(CTCs)监测等有着特殊的技术优势。本文简要介绍了单细胞测序的技术流程及数据处理并回顾目前单细胞测序在CRC应用进展, 从而为了解CRC发生及肿瘤进展过程中的肿瘤内异质性、深入理解CTCs的应用与寻找新的药物作用靶点提供参考。     

LC-MS联合单细胞测序分析探讨参苓白术散治疗原发性骨质疏松症的作用机制

目的 利用液质联用技术（LC-MS）以及单细胞转录组分析,研究参苓白术散作用于原发性骨质疏松症（primary osteoporosis,POP）的潜在分子机制。方法 采用LC-MS鉴定参苓白术散的化学成分,并预测其作用靶点。同时,获取POP相关的转录谱以及单细胞测序数据,分析其差异表达基因（differentially expressed genes,DEGs）和细胞亚群,进一步对DEGs进行加权基因共表达网络分析（WGCNA）,得到POP的枢纽基因,并构建参苓白术散-POP核心靶点网络。最后,对化学成分及其核心靶点进行虚拟分子对接验证。结果 LC-MS鉴定筛选后共得到10个化学成分,并得到414个潜在作用靶点。差异化及WGCNA分析分别得到843个POP-DEGs和1 976个枢纽靶点,联合分析后得到19个核心靶点。此外,POP骨髓血样本中存在巨噬细胞、单核细胞、T细胞、B细胞等免疫细胞。富集结果表明核心靶点涉及细胞衰老、破骨细胞分化、MAPK信号通路等。最后,虚拟分子对接结果显示鉴定成分与POP核心靶点能形成稳定对接。结论 参苓白术散能通过多成分、多靶点、多细胞及多通路调控细胞衰老、免疫功能及破骨分化等作用治疗POP。     

Characterization of the heterogeneity of R3327 rat prostatic tumors derived from single-cell clones

Prostatic adenocarcinoma is characterized by cellular diversity, which is well demonstrated in the Dunning R3327 rat prostatic adenocarcinoma. This heterogeneity may arise from epigenetic influences, ie, cellular adaptation or selection, and/or from genetic changes. To investigate the question of genetic instability, four tissue culture cell lines were derived from single cells isolated from the uncloned late (UCL) passage of the Dunning R3327H prostate cell culture. Each of these clonally derived tissue cultures was injected into castrated and intact young adult male rats for tumor production. Uncloned early (UCE) and UCL passage tissue cultures were also propagated as solid tumors. Tumors and the cultures from which they were derived were examined for evidence of phenotypic and genetic changes using morphological and cytometric methods. Transmission and scanning electron microscopy revealed only slight differences among the cell cultures. A single population of diploid cells was demonstrated in each of the cell cultures by propidium iodide staining and subsequent flow cytometric measurement of DNA content/nucleus. Tumors of unicellular as well as multicellular origin exhibited extreme heterogeneity of histological features, both among animals as well as within a single tumor. Tumors were surveyed and tissue types were characterized and cataloged. Clone 3 was generally better differentiated than the others; tumors from castrated animals were better differentiated than those from intact animals. Flow cytometry revealed multiple hyperdiploid cell populations that were variable from one sample to another. We concluded that changes in genotype as well as phenotype occurred in the tumors derived from single cells. Some of these changes may have occurred in the cells while still in culture.     

Th2 predominance at the single-cell level in patients with IgA nephropathy.

BACKGROUND: Abnormalities of lymphocyte function have been reported to be involved in the pathogenesis of IgA nephropathy (IgA-N). The aim of this study was to investigate helper T (Th) predominance at the single-cell level, one of the abnormalities of lymphocyte function in IgA-N. METHODS: Using flowcytometry, we assessed the levels of circulating Th cells in IgA-N patients (n=30), and in normal individuals (n=30) based on the expression of intracellular Th1 cytokines for interleukin-2 (IL-2) and interferon-gamma (IFN-gamma), and of intracellular Th2 cytokines for IL-4, IL-10, and IL-13. Because the production of each cytokine had a specific time course, we examined cytokine synthesis at 3, 6, 9, and 12 h after stimulation. RESULTS: The percentages of IL-2-positive Th cells from IgA-N patients were significantly lower than in normal individuals at 6, 9, and 12 h, with the difference becoming greater with time. The number of IFN-gamma-positive Th cells in IgA-N patients was significantly lower than in normal individuals at 9 h, and the number of IFN-gamma-positive Th cells increased more at 12 h than at 3 h in both groups. IL-4 and IL-13 expression was increased in patients with IgA-N at 6 h compared with normal individuals. In IgA-N patients, the percentage of IL-10-positive Th cells was significantly higher than that in normal individuals at each time-point. CONCLUSION: A polarization toward Th2 response at the stimulated lymphocyte level may lead to immune abnormalities in IgA-N.     

宏基因组二代测序技术在脊柱感染病原微生物诊断中的应用

【摘要】 目的：探讨宏基因组二代测序（metagenomic next-generation sequencing,mNGS）技术在脊柱感染疾病中诊断病原微生物的能力和价值。方法：纳入2019年1月～2020年12月收治的46例疑似脊柱感染患者,其中男性26例,女性20例;年龄26～77岁（50.4±15.7岁）。获取患者的血液和病灶组织或脓液标本（CT引导下穿刺活检21例,开放手术25例）,行微生物培养、血清学检测、病理检查和mNGS检测后进行结果的比较分析。结果：46例标本均在48h内获得了mNGS结果,微生物培养、血清学检测、病理检查等传统实验室检测时间1~12d不等。经临床最终诊断共35例为脊柱感染,11例为非脊柱感染。35例脊柱感染患者中,化脓性细菌感染14例,结核感染5例,布鲁菌属感染4例,未明确病原微生物感染12例;11例非脊柱感染患者中,脊柱肿瘤3例,终板Modic改变4例,终板骨折1例,DISH病合并假关节1例,强直性脊柱炎合并假关节2例。通过mNGS检测发现了脆弱拟杆菌、微小微单胞菌、齿垢密螺旋体、贝纳柯克斯体等少见病原微生物。mNGS在种水平与临床最终诊断的一致性为82.61%（19/23）,在属水平的一致性为95.65%（22/23）。微生物培养阳性率为42.86%（15/35）,mNGS检测阳性率为94.29%（33/35）,二者存在统计学差异（P＜0.01）。mNGS检测的敏感度为91.43%,特异度为90.91%,阳性预测值96.97%,阴性预测值76.92%。结论：宏基因组二代测序技术检测病原微生物快速、高效、准确,在脊柱感染疾病的诊疗中有较高的诊断价值。     

Correlation maps allow neuronal electrical properties to be predicted from single-cell gene expression profiles in rat neocortex

The computational power of the neocortex arises from interactions of multiple neurons, which display a wide range of electrical properties. The gene expression profiles underlying this phenotypic diversity are unknown. To explore this relationship, we combined whole-cell electrical recordings with single-cell multiplex RT-PCR of rat (p13-16) neocortical neurons to obtain cDNA libraries of 26 ion channels (including voltage activated potassium channels, Kv1.1/2/4/6, Kvbeta1/2, Kv2.1/2, Kv3.1/2/3/4, Kv4.2/3; sodium/potassium permeable hyperpolarization activated channels, HCN1/2/3/4; the calcium activated potassium channel, SK2; voltage activated calcium channels, Caalpha1A/B/G/I, Cabeta1/3/4), three calcium binding proteins (calbindin, parvalbumin and calretinin) and GAPDH. We found a previously unreported clustering of ion channel genes around the three calcium-binding proteins. We further determined that cells similar in their expression patterns were also similar in their electrical properties. Subsequent regression modeling with statistical resampling yielded a set of coefficients that reliably predicted electrical properties from the expression profile of individual neurons. This is the first report of a consistent relationship between the co-expression of a large profile of ion channel and calcium binding protein genes and the electrical phenotype of individual neocortical neurons.     

Vascular heterogeneity in the kidney

Blood vessels and their endothelial lining are uniquely adapted to the needs of the underlying tissue. The structure and function of the vasculature varies both between and within different organs. In the kidney, the vascular architecture is designed to function both in oxygen/nutrient delivery and filtration of blood according to the homeostatic needs of the body. Here, we review spatial and temporal differences in renal vascular phenotypes in both health and disease.     

基因直接测序分析HBV多聚酶区基因突变与基因分型

HBV是一种嗜肝DNA病毒,其基因组为3200bp的双链DNA分子,按照全基因序列的异质性,HBV可分为A-H等8个基因型。HBV的变异率极高,而且具有很高的复制率,在由共价闭合环状DNA反转录过程中因HBV聚合酶和反转录酶缺乏校正活性,因此不能去除错误掺入的碱基。HBV常引起慢性持续感染,在长期反复感染和复制中发生变异,并受人体免疫应答、抗病毒药物等环境因素的影响。核苷（酸）类似物拉米夫定和替比夫定等抗病毒药物的作用靶点均为HBV聚合酶和反转录酶,