中药火麻仁基原植物大麻的TIFY基因家族鉴定及功能分析

目的:从全基因组层面对大麻(Cannabis sativa)TIFY基因家族进行鉴定与功能分析,为解析大麻TIFY家族基因功能及其对大麻素等次生代谢产物生物合成的调控机理奠定基础。方法:采用已有大麻基因组数据,通过NCBI,PlantTFDB,MEME及TBtools等生物信息学分析工具,鉴定大麻中的TIFY家族基因,并对其编码蛋白的理化性质、系统进化、基因结构、染色体定位及组织差异表达模式等进行分析和可视化作图。结果:该研究共鉴定到大麻TIFY家族基因成员14个(CsTIFY1～CsTIFY14),分属于TIFY,JAZ,ZML和PPD共4个亚家族,其核酸序列长度为365～1 369 bp,编码氨基酸长度为118～442 aa,等电点为4.64～9.96。14个CsTIFYs不均匀的分布在8条染色体上,亚细胞定位预测其蛋白均定位细胞核中。CsTIFYs基因的启动子区具有多种非生物胁迫响应的顺式作用元件,可参与植物的不同生长发育和非生物胁迫调控。转录组表达热图分析表明,CsTIFYs在不同大麻品种的雌花及同一品种的花、苞片、茎、叶中均存在表达差异。结论:该研究从全基因组水平鉴定到大麻CsTIFY家族转录因子14个,并对其结构特点和表达模式进行研究,预测大麻CsTIFYs可能在大麻的JA信号转导通路、非生物胁迫及大麻素生物合成中发挥重要调控作用,将对大麻中TIFY家族的基因功能研究以及大麻优质品种选育提供科学参考。     

基于网络药理学和分子对接技术探讨生脉注射液抗新型冠状病毒肺炎的作用机制

目的采用网络药理学与分子对接技术探讨生脉注射液的活性成分和治疗新型冠状病毒肺炎(COVID-19)的潜在作用机制。方法利用TCMSP及BATMAN-TCM数据库筛选生脉注射液的活性化合物,通过TCMSP及Targetnet在线数据库预测作用靶点,通过Cytoscape3.7.1构建活性成分-作用靶点网络图;在GeneCards及OMIM数据库中以"coronavirus pneumonia"为关键词搜索冠状病毒肺炎相关疾病靶点,与生脉注射液化合物靶点进行交集筛选出共同靶点作为研究靶点,将共同靶点导入STRING数据库获取数据后在Cytoscape 3.7.1软件中构建蛋白质-蛋白质相互作用网络图;利用R语言进行GO(gene ontology)功能、KEGG(Kyoto encyclopedia of genes and genomes)通路富集分析,预测其作用机制,并构建"成分-靶点-通路"网络图;通过DiscoveryStudio 2.5软件对关键靶点进行分子对接分析。结果生脉注射液筛选得到22个活性化合物,分别为邻苯二甲酸二辛酯、β-谷甾醇、当归酰基戈米辛O、戈米辛A、戈米辛R、五味子丙素、内南五味子酯乙、长南酸、南五味子内酯、香蒲木脂素B、新杜松烷酸A、新杜松烷酸B、新杜松烷酸C、新南五味子木脂宁、五味子内酯A、五味子内酯E、五味子酸、尿苷、薯蓣皂苷元、鸟嘌呤核苷、N-反式阿魏酰酪胺、豆甾醇。相应作用靶点224个,与COVID-19的共同靶点16个,分别为CASP3、CASP8、PTGS2、BCL2、BAX、PRKCA、PTGS1、PIK3CG、F10、NOS3、DPP4、NOS2、TLR9、ACE、ICAM1、PRKCE,关键靶点涉及CASP3、PTGS2、NOS2、NOS3、ICAM1。GO功能富集分析得到生物过程(BP)条目771个,细胞组成(CC)条目11个,分子功能(MF)条目79个。KEGG通路富集分析筛选得到67条(P0.05)信号通路,主要涉及糖尿病并发症AGE-RAGE信号通路、凋亡通路、P53信号通路、小细胞肺癌通路等。分子对接结果显示与关键靶点对接较好的成分有五味子内酯E、豆甾醇、N-反式阿魏酰酪胺。结论生脉注射液中的活性化合物五味子内酯E、豆甾醇、N-反式阿魏酰酪胺等能作用于CASP3、PTGS2、NOS2、NOS3等靶点调节多条信号通路发挥抗炎、免疫调节、抗休克、增加血氧饱和度等作用,从而可能发挥对COVID-19的治疗作用。     

A prognostic signature based on immune-related genes for cervical squamous cell carcinoma and endocervical adenocarcinoma

Cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC) is the fourth commonest female malignancy worldwide. CESC progresses in immune-microenvironment mainly composed of infiltrating immune and stromal cells. Here, we performed an integrated analysis incorporating the expression profiles from the Cancer Genome Atlas (TCGA) database and scores of immune and stromal cells calculated by Estimation of Stromal and Immune cells in Malignant Tumours using Expression data (ESTIMATE) algorithm. A two-gene signature (CD1C and CD6 genes) was established to predict the prognosis of CESC. Based on this signature, patients were divided into the high- and low-risk groups, and this signature showed good prognostic performance according to the results of Kaplan-Meier analysis and receiver operating characteristic (ROC) analysis in train set and two validation sets. A nomogram was built for evaluating the clinical applicability of this signature. In addition, based on Tumor Immune Estimation Resource (TIMER) database, 2 hub genes showed negative correlations with tumor purity and positive correlations with infiltrating levels of immune filtrating cells. What’s more, we propose new treatment strategies for the two prognostic subtypes. Low- risk patients were found presenting with a higher level of immune checkpoint molecules and showing higher immunogenicity in immunophenoscore (IPS) analysis, which indicated a better response for immunotherapy. Meanwhile, estimated by Genomics of Drug Sensitivity in Cancer (GDSC) database, the high-risk patients showed sensitive responses to five chemotherapy drugs. Finally, 10 candidate small-molecule drugs for CESC were defined. In summary, the CD1C-CD6 signature can accurately predict the prognosis of CESC.     

拔罐治疗非特异型颈椎病患者颈椎功能障碍Meta分析

目的:探究拔罐治疗非特异型颈椎病患者颈椎功能障碍的临床疗效及不同拔罐方式、治疗时间应用的差异性。方法:计算机网络检索Pub Med、Embase、Web of science外文数据库及中国知网(CNKI)、维普全文(VIP)、万方、中国生物医学等中文数据库自建库起至2018年3月1日公开发表的关于拔罐治疗非特异型颈椎病涉及颈椎功能障碍指数(Neck Disability Index,NDI)的临床随机对照研究(RCT)。由2名研究员独立筛选文献并提取资料。采用Rev Man 5.3软件进行Meta分析。结果:共纳入7项研究、全部为RCT研究,采用固定效应模型合并效应量加权均数差(WMD)对文献进行分析,显示拔罐可以改善非特异型颈椎病患者的颈椎功能障碍,差异有统计学意义(P0.00001);联合拔罐与单纯火罐比较临床疗效差异无统计学意义(P=0.37);短期(≤4周)与长期( 4周)比较差异无统计学意义(P=0.27)。结论:拔罐可以显著改善非特异型颈椎病患者的颈椎功能障碍,疗效与拔罐方式、治疗时间无相关性。     

丹参F3'5'H基因克隆及其序列分析

目的:为了验证丹参类黄酮-3′,5′-羟基化酶(Flavonoid 3′,5′-hydroxylase,F3′5′H)基因与丹参花色表型的相关性,本研究克隆并分析了紫花丹参99-3株系和一些白花丹参中的F3′5′H基因。方法:本研究通过提取紫花丹参和白花丹参中的总RNA,再将其反转录得到cDNA,以此cDNA为模板,利用PCR方法扩增获得F3′5′H基因全长序列。再利用生物信息学分析方法,分析了该基因编码蛋白质的理化性状、结构域、系统进化等特点,并预测了该蛋白质的亚细胞定位、跨膜区等;利用实时定量PCR方法检测了该基因的表达特异性;利用毛状根遗传转化方法获得了该基因的过表达阳性毛状根株系。结果:F3′5′H基因全长1551 bp,编码516个氨基酸,相对分子质量为57.4 kDa。该基因在丹参花中高丰度表达。在42份(紫花25份;白花17份)丹参样品中发现一个单核苷酸多态性(Single Nucleotide Polymorphisms,SNP)位点在紫花和白花丹参中呈现与花色相关的稳定变化。系统发育分析表明丹参F3′5′H与美女樱的F3′5′H具有较高序列同源性。获得了过表达F3′5′H的毛状根株系。结论:本研究在丹参中克隆到F3′5′H基因,对其序列进行了分析,为进一步研究丹参F3′5′H基因的功能奠定基础。     

The effects of grape seed extract on glycemic control,serum lipoproteins,inflammation, and body weight: A systematic review and meta‐analysis of randomized controlled trials

The aim of this systematic review and meta‐analysis was to analyze the effects of grape seed extract (GSE) on glycemic control and serum lipoproteins, inflammation and body weight. Two independent authors systematically searched online databases including EMBASE, Scopus, PubMed, Cochrane Library, and Web of Science from inception until May 30, 2019. Cochrane Collaboration risk of bias tool was applied to assess the methodological quality of included trials. The heterogeneity among the included studies was assessed using Cochrane's Q test and I‐square (I²) statistic. Data were pooled using a random‐effects model and weighted mean difference (WMD) was considered as the overall effect size. Fifty trials were included in this meta‐analysis. Pooling effect sizes from studies demonstrated a significant decrease in fasting plasma glucose (FPG) (WMD): ?2.01; 95% confidence interval (CI): ?3.14, ?0.86), total cholesterol (TC; WMD: ?6.03; 95% CI: ?9.71, ?2.35), low‐density lipoprotein (LDL) cholesterol (WMD: ?4.97; 95% CI: ?8.37, ?1.57), triglycerides (WMD: ?6.55; 95% CI: ?9.28, ?3.83), and C‐reactive protein (CRP) concentrations (WMD: ?0.81; 95% CI: ?1.25, ?0.38) following GSE therapy. Grape seed did not influence HbA1c, HDL cholesterol levels, and anthropometric measurements. This meta‐analysis demonstrated that GSE intake significantly reduced FPG, TC, LDL cholesterol, triglycerides, and CRP levels.     

我国轻度认知障碍研究热点分析及展望

背景　随着人口老龄化进程的加快，包括痴呆在内的老年人健康问题日益突出。轻度认知障碍作为阿尔茨海默病的临床前阶段，早已成为研究者关注的热点。但是目前尚缺乏对我国轻度认知障碍研究热点的整理与分析。目的　分析2008-2018年我国轻度认知障碍研究的热点，了解该领域的现状和发展趋势。方法　2019年6月，以“轻度认知障碍”为关键词，采用主题检索的方式，在中国期刊网全文数据库（CNKI）中进行检索，检索时间为2008-2018年，文献来源类别限定为SCI来源文献、EI来源期刊、核心期刊、中文社会科学引文索引（CSSCI）以及中国科学引文数据库（CSCD）。采用中医药数据关联分析平台（XMiner软件）、SPSS 22.0统计学软件、Excel软件对关键词进行词频分析和共词聚类分析。结果　最终纳入符合标准的文献413篇，获得高频关键词41个（g=41）。通过共词聚类分析总结出2008-2018年我国轻度认知障碍的7个研究热点，分别为轻度认知障碍的病因学研究、神经心理评估及筛查研究、诊断及分类研究、干预研究、流行病学研究、精神行为症状研究、生物标志物及中医证候研究。结论　2008-2018年我国轻度认知障碍的研究热点分别为轻度认知障碍的病因学研究、神经心理评估及筛查研究、诊断及分类研究、干预研究、流行病学研究、精神行为症状研究、生物标志物及中医证候研究，该分析结果有助于广大卫生服务人员了解该领域的现状和发展趋势，且对其今后的研究方向有一定的指导作用。     

基于灰色关联度分析和聚类分析的3种滇产重楼抗肝癌作用谱效关系探索

目的:研究长柱重楼、滇重楼和南重楼的HPLC指纹图谱与其抗肝癌作用的谱效关系,为明确重楼抗肝癌作用的物质基础提供实验依据。方法:采用HPLC建立3种重楼提取物的指纹图谱,流动相乙腈(A)-水(B)梯度洗脱(0～10 min,20%A; 10～20 min,20%～25%A; 20～30 min,25%～30%A; 30～40 min,30%～35%A; 40～50 min,35%～40%A; 50～60 min,40%A; 60～75 min,40%～45%A; 75～80 min,45%～60%A),流速0. 9 m L·min~(-1),检测波长203 nm;利用噻唑蓝(MTT)比色法测定3种重楼提取物对肝癌HepG2细胞的增殖抑制作用,计算半数抑制浓度(IC_(50));运用聚类分析(HCA)和灰色关联度分析(GRA)研究3种重楼指纹图谱和抗肝癌作用的关系,找出对抗肝癌作用贡献较大的成分。结果:在3种重楼的HPLC指纹图谱中,确定其中11个色谱峰为共有峰。作用时间72 h时长柱重楼、滇重楼、南重楼的IC_(50)分别为148. 33,178. 87,208. 09 mg·L~(-1),其中长柱重楼的抗肝癌活性最强。灰色关联度结果显示,滇重楼共有峰中关联度较高的为1～10号峰,长柱重楼共有峰关联度较高的为1～7号峰,南重楼共有峰中关联度较高的为1～4,6～10,N1号峰,与IC_(50)关联度均0. 7。各重楼变量的聚类分析结果显示,可与IC_(50)聚为一类的色谱峰的关联度均 0. 7。结论:建立了3种重楼的HPLC指纹图谱,重复性良好。3种重楼中的1～4,6和7号色谱峰对抗肝癌药效贡献最大。     

The effect of nettle (Urtica dioica) supplementation on the glycemic control of patients with type 2 diabetes mellitus: A systematic review and meta‐analysis

Type 2 diabetes mellitus (T2DM) is a major health problem, worldwide, that is associated with increased morbidity and mortality. Several randomized controlled clinical trials (RCTs) have investigated the effect of nettle (Urtica dioica) supplementation on markers of glycemic status in patients with T2DM, with conflicting results. Therefore, the present study assessed the effect of nettle on some glycemic parameters in patients with T2DM. A comprehensive search was conducted in PubMed, Scopus, Cochrane Library, and Web of Science, from database inception up to June 2019, to identify RCTs investigating the effect of nettle supplementation on glycemic markers, including fasting blood sugar (FBS) concentrations, insulin levels, homeostasis model assessment‐estimated insulin resistance index, and glycosylated hemoglobin percentage in adults with T2DM. The Cochrane Collaboration tool was used to assess the methodological quality of the included studies. Results of this meta‐analysis were reported based on the random effects model. Eight RCTs, comprising 401 participants, were included in the present systematic review and meta‐analysis. Based on the Cochrane Collaboration risk of bias tool, five studies were considered as good quality, one was fair, and two studies were poor, respectively. The results of the meta‐analysis revealed a significant reduction in FBS concentrations (weighted mean difference [WMD]: ?18.01 mg/dl, 95% confidence interval [CI]: ?30.04 to ?5.97, p < .001, I² = 94.6%) following nettle supplementation. However, no significant reduction was observed in insulin levels (WMD: 0.83 Hedges' g, 95% CI: ?0.26 to 1.92, p = .13, I² = 89.4%), homeostasis model assessment‐estimated insulin resistance index (WMD: ?0.22, 95% CI: ?0.83 to 0.40, p = .49, I² = 69.2%), or glycosylated hemoglobin percentage (WMD: ?0.77%, 95% CI: ?1.77 to 0.22, p = .12, I² = 83.0%). The findings of the present study suggest that nettle supplementation may be effective in controlling FBS for T2DM patients. However, further studies are needed to confirm the veracity of these results.