单细胞测序数据的智能解析与数据库

随着测序技术发展,单细胞测序已经成为发育医学研究使用的热点技术。本文概述近年来单细胞测序技术在发育医学研究上的应用范围,整合单细胞测序数据与分析的重要性,解析近年来发布的单细胞测序数据库和分析工具,并用表图展现各平台的访问地址及功能特点。     

去唾液酸糖蛋白受体H1亚单位基因片段的克隆表达及其检测自身免疫性肝炎的价值

Objective To clone and express the human asialoglycoprotein receptor(ASGPR) H1 subunit, purify and identify the immunoreactivity of the recombinant protein, and establish the enzyme linked immunosorbent assay (ELISA) to detect anti-ASGPR antibodies in diagnosis of autoimmune hepatitis. Methods The CRDHI cDNA (435 bp) was subcloned into eukaryotic vector PEGH, and the recombinant protein expression was induced by D (+)-Galactose. The recombinant CRDH1 was purified with Glutathione Sepharose 4B, and its immunoreactivity was identified by SDS-PAGE and western blot as well as MALDI-TOF. ELISA was established to detect the anti-ASGPR antibodies in serum samples of 45 patients with AIH, 30 patients with SLE, 30 patients with RA, 10 patients with SS and 30 normal controls. Results The sequencing of recombinant plasmid showed the CRDH1 gene was successfully inserted to the eukaryotic expression vector with correct sequence and open reading frame. The fusion protein showed a molecular weight of 42 500 Da on SDS-PAGE gel and confirmed to be the human ASGPR by MALDI-MS through peptide mass fingerprint analysis with Mascot in human protein database. It shared 98. 34% homology with ASGPR H1 subunit. Western blot analysis showed that the fusion protein had the same immunoreactivity as human ASGPR. The results of ELISA indicated that the positive rate of anti-ASGPR was 35.6% ( 16/45 ), but the ELISA was negative in other control. There was significant difference of positivity of the autoantibodies between AIH and non-AIH controls (χ2 = 31.85,P < 0. 01 ). Conclusions The human plasmid containing ASGPR is successfully clone into Saccharomyces cerevisiae Y258. The recombinant autoantigen owns good antigenicity and specificity. ELISA established with the purified protein shows good specificity for diagnosis of AIH.     

含盐酸二甲双胍联合方案治疗再生障碍性贫血的短期疗效观察

目的采用临床生物信息学方法筛选治疗再生障碍性贫血(AA)的新型药物并评估其临床疗效。方法首先建立AA的人类全基因组表达谱,与药物基因组学数据库进行相似性分析,筛选可能有效的药物。纳入难治性AA患者[接受过免疫抑制剂和(或)雄激素治疗时间超过6个月无效]作为研究对象评估疗效,6个月后评估临床疗效和不良反应。结果临床生物信息学筛选结果显示盐酸二甲双胍可能对AA有治疗作用。纳入43例难治性AA患者(其中15例为重型AA),采用包含盐酸二甲双胍、环孢素A(CsA)和司坦唑醇的联合方案进行治疗,结果显示:27例输血依赖患者在治疗6个月后全部(100%)停止输血;40例贫血患者中,37例(92.5%)血红蛋白完全恢复正常;30例血小板低于20×109/L的患者中,28例(93.3%)升至50×109/L以上;35例白细胞低于2.5×109/L的患者中,31例(88.6%)升至3.5×109/L以上。40例贫血患者中,1例出现肾功能异常,停用环孢素A后恢复正常;18例出现转氨酶升高,加用保肝药物并减少司坦唑醇用量后恢复正常。所有患者均未出现低血糖反应。结论盐酸二甲双胍、环孢素A和司坦唑醇联合方案治疗难治性AA有效。     

深度学习在医学影像学领域应用研究进展

阐述深度学习方法、原理和主要模型结构,以脑部疾病和乳腺癌为例分析深度学习在国内医学影像学领域的应用情况,总结其局限性,包括图像获取难度高、数据缺乏完整性、数据采集标准化不足及图像像素分辨率不高等方面。     

信号蛋白3A对成肌细胞功能的影响及其转录调控机制

目的 探讨信号蛋白3A（Sema3A）对成肌细胞功能的影响及其转录调控机制。方法 以1.0、0.1、0.01 μg/ml重组Sema3A蛋白作用成肌细胞,并设置对照组,采用动态活细胞成像及划痕实验检测细胞的增殖与迁移能力,RT-qPCR检测成肌分化因子的表达。对1.0 μg/ml重组Sema3A蛋白干预的成肌细胞及对照组细胞进行转录组测序,筛选差异表达基因并进行GO富集分析,对部分差异表达基因通过RT-qPCR进行转录水平验证。结果 IncuCyte细胞成像计数显示,Sema3A可促进成肌细胞增殖,其中1.0 μg/ml Sema3A组细胞计数较对照组升高17.36%（_P<0.001）。划痕实验结果显示,Sema3A可使成肌细胞迁移能力明显增强,其中1.0 μg/mlSema3A组细胞划痕愈合率较对照组升高69.66%（_P<0.001）。RT-qRCR检测结果显示,Sema3A可上调部分成肌分化基因的表达,其中1.0 μg/ml Sema3A可使_Myf5、_MyoG基因表达分别升高37.4...     

神经丝蛋白与帕金森病的相关性研究

目的研究神经丝蛋白(NFs)作为帕金森病(PD)外周生物标记物的可能性及临床意义。方法用酶联免疫吸附方法检测23例PD患者、30例急性脑梗死患者和29名健康对照者血清NFs水平并进行比较。分析PD患者病程和血清NFs水平的相关性。结果 PD患者和急性脑梗死患者血清NFs水平均明显高于健康对照(P0.05),而PD患者和急性脑梗死患者之间血清NFs水平差异无统计学意义(P0.05)。PD患者血清NFs水平和病程具有相关性(r=0.739,P0.05)。结论 PD患者疾病过程中存在轴索损伤,且轴索损伤严重程度可能随病程延长持续加重;PD患者血清NFs水平改变无疾病特异性,不能作为理想的PD外周生物标记物。     

去唾液酸糖蛋白受体H1亚单位基因片段的克隆表达及其检测自身免疫性肝炎的价值

Objective To clone and express the human asialoglycoprotein receptor(ASGPR) H1 subunit, purify and identify the immunoreactivity of the recombinant protein, and establish the enzyme linked immunosorbent assay (ELISA) to detect anti-ASGPR antibodies in diagnosis of autoimmune hepatitis. Methods The CRDHI cDNA (435 bp) was subcloned into eukaryotic vector PEGH, and the recombinant protein expression was induced by D (+)-Galactose. The recombinant CRDH1 was purified with Glutathione Sepharose 4B, and its immunoreactivity was identified by SDS-PAGE and western blot as well as MALDI-TOF. ELISA was established to detect the anti-ASGPR antibodies in serum samples of 45 patients with AIH, 30 patients with SLE, 30 patients with RA, 10 patients with SS and 30 normal controls. Results The sequencing of recombinant plasmid showed the CRDH1 gene was successfully inserted to the eukaryotic expression vector with correct sequence and open reading frame. The fusion protein showed a molecular weight of 42 500 Da on SDS-PAGE gel and confirmed to be the human ASGPR by MALDI-MS through peptide mass fingerprint analysis with Mascot in human protein database. It shared 98. 34% homology with ASGPR H1 subunit. Western blot analysis showed that the fusion protein had the same immunoreactivity as human ASGPR. The results of ELISA indicated that the positive rate of anti-ASGPR was 35.6% ( 16/45 ), but the ELISA was negative in other control. There was significant difference of positivity of the autoantibodies between AIH and non-AIH controls (χ2 = 31.85,P < 0. 01 ). Conclusions The human plasmid containing ASGPR is successfully clone into Saccharomyces cerevisiae Y258. The recombinant autoantigen owns good antigenicity and specificity. ELISA established with the purified protein shows good specificity for diagnosis of AIH.     

西双版纳州健康人群血清丙氨酸氨基转移酶参考区间调查

目的建立西双版纳州健康人群血清丙氨酸氨基转移酶(ALT)丙酮酸氧化酶法参考区间。方法对3167名健康者血清ALT活性在不同性别、不同年龄组间的分布进行统计分析。结果ALT活性男性明显高于女性(P<0.001)。ALT参考区间上限:男性为42 U/L,女性为37 U/L。男性血清ALT活性在30~59岁组较高(其中40~49岁组最高),<20岁组最低;ALT参考区间上限:<20岁为34 U/L,20~29岁为46 U/L,30~59岁为47 U/L,≥60岁为39 U/L。女性≥40岁者血清ALT活性较高(其中50~59岁组最高),≤29岁者较低;ALT参考区间上限:≤29岁为34 U/L,30~39岁为35 U/L,≥40岁为40 U/L。结论ALT活性在不同性别、不同年龄组间有明显差异,各实验室应按不同性别、不同年龄建立本地区、本实验室血清ALT参考区间。     

基质辅助激光解析/电离飞行时间质谱法检测高危型人乳头瘤病毒基因型及临床应用

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