ITS2���м������ԭҩ�����ٲ�

??OBJECTIVE To identify 93 samples of Ilex asprella(Hook. et Arn.) Champ. ex Benth. and its related species based on ITS2 barcode. METHODS The total genomic DNAs were extracted and the ITS2 regions were amplified and sequenced. The genetic distances were computed using MEGA5.0 in accordance with the Kimura 2-parameter (K2P) model. The neighbor-joining (NJ) phylogenetic tree and BLAST method were used to assess the efficiency of sequence identification. RESULTS The length of ITS2 sequence of I. asprella was 241 bp and the average GC content was 63.4%. The intra-specific genetic distances of I. asprella were 0-0.030 and the minimum inter-specific of I. asprella and its closely related species was 0.043. Furthermore, I.asprella formed into a single branch with high bootstrap value in NJ tree. The identification efficiency of ITS2 barcode using BLAST was 100%. CONCLUSION The ITS2 region can accurately and stably distinguish I.asprella from its closely related species and adulterants.It provides a new method to ensure the clinical safety of traditional Chinese medicines.     

GLP-1���弤�����ཱུ��ҩ��Ѫ�ܰ�ȫ�Ե�Meta����

??OBJECTIVE To evaluate the cardiovascular safety of glucagon-like peptide-1 receptor agonists (GLP-1 RA) for diabetes systematically. METHODS Medline, Embase, ClinicalTrails.gov, Cochrane Liabrary, CBM, and CNKI were retrieved to collect all the randomized controlled trials (RCT) with a duration of at least 24 weeks, comparing a GLP-1RA with a non-GLP-1 RA agent in type 2 diabetes, and then a Meta-analysis was performed with RevMan5.3 software. RESULTS Fifty-one RCTs, invovling 26 140 individuals with type 2 diabetes, were included in the analysis. The difference in the incidence of major adverse cadiovascular events (MACE) between GLP-1RA and comparators did not reach statistical significance, and the odds ratio was 0.78 (0.57, 1.08), although a favorable trend was observed; while in comparisons with placebo, the incidence of MACE was significantly decreased and the odds ratio was 0.5 (0.28, 0.87); compared with comparators, a significant decrease of the incidence of all-cause and cardiovascular mortality were detected and the odds ratio were 0.53 (0.27, 0.93) and 0.38 (0.16, 0.90), respectively. As for the cardiovascular risk factors, GLP-1 RA significantly decreased the HbA1c, weight, TG, LDL, SBP, but increased the heart rates. CONCLUSION GLP-1 RA may have a protective effect on cardiovascular diseases, compared with the comparators, especially compared with placebo.     

�ȸ�Ѫ��������1���弤��������2�����򲡺ϲ��Ǿƾ���֬������Ч�Ե�ϵͳ���ۺ�Meta����

??OBJECTIVE To evaluate the efficacy of glucagon-like peptide-1 receptor agonists (GLP-1RA) on nonalcoholic fatty liver disease (NAFLD) in patients with type 2 diabetes mellitus (T2DM). METHODS The randomized controlled trials (RCTs) of GLP-1 RA, placebo, and anti-diabetes drugs in the treatment of NAFLD in patients with T2DM were collected from PubMed, Embase, CNKI, Wangfang database, VIP, and CBM. The trials were evaluated for the quality and selected, and the RESULTS of the studies were reviewed and analyzed using RevMan 5.2 software. RESULTS Four RCTs were included, involving 154 patients. The Meta-analysis showed that compared with the control group, GLP-1RA could significantly improve the ALT [MD:-8.36,95%CI(-13.41-3.31), P=0.001], HbA1c [MD:-0.43%,95%CI(-0.73-0.31), P=0.005], FBG [MD:-0.71%, 95%CI(-1.39-0.03),P=0.04],BMI [MD:-1.38%, 95%CI(-2.18-0.58), P=0.000 8], TG [MD:-0.49%, 95%CI(-0.82-0.16), P=0.004]. CONCLUSION GLP-1 RA can obviously improve the metabolic index of patients with NAFLD and T2DM. Given the quality and quantity of the literature, large RCTs are still needed in the future.     

����ITS2���е�ҩ�øʲ���Ե��ϵ�����о�

??OBJECTIVE To obtain the genetic information of 181 domestic and foreign licorice samples by determining the internal transcribed spacer sequence 2 (ITS2) in order to clarify their phylogenetic relationships and differentiate the medicinal licorice species from the non-medicinal ones. METHODS Genomic DNA was first extracted from licorice samples of different origins and habitats. Then, the ITS2 sequences were amplified using PCR technique. Next, the ITS2 sequences were subjected to sequence analyses and neighbor-joining(NJ) tree analysis. Finally, the analytical method based on ITS2 DNA barcodes was established for the licorice species discrimination. The method was then applied to the phylogenetic analysis among the suspected domestic mutants, foreign licorice samples and domestic medicinal licorices, in addition to the discriminations of the medicinal and non-medicinal licorices. RESULTS The medicinal licorice could be explicitly separated from the two commonly encountered non-medicinal licorice species either by using ITS2 sequence analysis or NJ tree analysis; and the suspected domestic mutants and foreign licorice samples converged with the medicinal licorice to form one monophyletic branch in the NJ tree analysis, which indicated unambiguously conspecifics. CONCLUSION This study establishes a method for discrimination of medicinal and non-medicinal licorices based on ITS2 barcodes; and by the advantage of this method, the conspecific relationships among the suspected domestic mutants, foreign licorice samples and Chinese medicinal licorice are clarified. Thus a theoretical reference to the expansion of medicinal licorice resources and assurance of the safety and efficacy of the clinical licorice medications are provided.     

DPP-4���Ƽ���GLP-1���弤�����Ƚ�����2��������Ч�ԺͰ�ȫ�Ե�Meta����

??OBJECTIVE To systematically review the efficacy and safety of dipeptidyl peptidase-4(DPP-4) inhibitors versus glucagon like peptide-1(GLP-1) receptor agonists for type 2 diabetes mellitus(T2DM). METHODS Databases including EMbase,PubMed, The Cochrane Library, Clinical Trial, CBM,CNKI and WanFang Data, were searched electronically for randomized controlled trials (RCTs) of DPP-4 inhibitors versus GLP-1 receptor agonists for T2DM up to December 2015. Two reviewers independently screened literatures according to the inclusion and exclusion criteria, extracted data, and assessed the methodological quality of the included studies. Then Meta-analysis was performed using RevMan 5.3 soft ware. RESULTS A total of 13 RCTs were included. The RESULTS of Meta-analysis showed that compared with DPP-4 inhibitors, GLP-1 receptor agonists were more effective in reducing the levels of fasting plasma glucose [MD=0.93, 95%CI(0.48,1.38), P??0.000 1] and glycated hemoglobin [MD=0.53,95%CI(0.34,0.73), P??0.000 01] and BMI [MD=1.53,95%CI(0.83,2.22),P??0.001]. However, DPP-4 inhibitors were more effective than GLP-1 receptor agonists in the reducing the 2-hour postprandial blood glucose level. And GLP-1 receptor agonists were more prone to cause gastrointestinal adverse reactions than DPP-4 inhibitors [RR=0.44,95% CI (0.33, 0.59), P<0.000 1]. CONCLUSION GLP-1 receptor agonists are superior to DPP-4 inhibitors in controlling the fasting plasma glucose and glycated hemoglobin levels and reducing the body weight of T2DM patients, while DPP-4 inhibitors have better efficacy in reducing 2-hour postprandial blood glucose level, with better tolerability.     

Ҷ����psbA-trnH���м���ҩʳͬԴ�����ҩ��

??OBJECTIVE To introduce DNA barcoding based on plastid intergenic region psbA-trnH to authenticate medicinal and edible materials Cinnamon cassia to promote the standard practice of medication and foods. METHODS The genomic DNA of eight Cinnamomum species were extracted successfully. The Kimura 2-parameter (K2P) distances were calculated and phylogenetic tree was constructed using software MEGA 5.0. RESULTS The sequence length of the psbA-trnH of Cinnamomum cassia was 368 bp. The psbA-trnH intra-specific genetic distance of C. cassia was highly conservative,which was shorter than the interspecific ones. The NJ tree indicated that Cinnamomi Cortex could be distinguished from its adulterants obviously. CONCLUSION The psbA-trnH intergenic spacer can be used to distinguish Cinnamomi Cortex from its adulterants accurately     

����AGREE ���2������ָ�Ϻ͹�ʶ��������

??OBJECTIVE To evaluate the quality of now available clinical guidelines and consensus statements on type 2 diabetes. Based on this assessment, the comparison of guidelines and consensuses is made to give some suggestions on the METHODS of making these documents. METHODS PubMed, Embase, CNKI, Wanfang, SinoMed and guideline websites were systematically searched. Literatures were screened according to defined criteria for including. AGREE ?? was used to instrument assess the quality of the guidelines and consensuses. RESULTS The result showed that the quality of included guidelines is various, but the overall quality of guidelines is still higher than consensuses. Guidelines got high scores in scope and purpose domain and clarity of presentation domain, but low scores in rigor of development domain and applicability domain. Consensus statements got low scores in all 6 domains. CONCLUSION The main reason for the difference is consensuses have poor methodology. Rapid advice guidelines were suggested as an alternative for consensus statements.     

��ͬ�ӹ����κ�����ɷֵĺ����ⶨ��TOPSIS����

??OBJECTIVE To establish a QTRAP-UPLC-MS/MS method for simultaneous determination of 10 kinds of nucleosides and nucleobases in Scrophulariae Radix processed by different methods, and investigate the influences of different processing < br/>METHODS on contents of nucleosides and nucleobases. METHODS The contents of 10 kinds of nucleosides and nucleobases in Scrophulariae Radix processed with different METHODS were simultaneously determined by QTRAP-UPLC-MS/MS, and TOPSIS analysis was used for comprehensive evaluation of nucleosides and nucleobases. RESULTS The contents of guanosine,uridine, adenosine and uracil were relatively high in Scrophulariae Radix.There were differences in contents of nucleosides and nucleobases of Scrophulariae Radix processed with different methods, and the samples processed by method of ??sweating?? had better quality. CONCLUSION This study provides a scientific basis for investigation on the influence of processing METHODS on contents of nucleosides and nucleobases in Scrophulariae Radixand screening of suitable processing method for Scrophulariae Radix.     

��ҩ���Ƥ�����αƷ��DNA����������о�

??OBJECTIVE To choose suitable barcodes for Draconis Sanguis and find the new method to identify Draconis Sanguis. METHODS The genomic DNAs from 41 samples were extracted. Draconis Sanguis samples were amplified and sequenced based on seven candidate DNA barcodes . The obtained sequences were assembled using the CodonCode Aligner V 4.2. The Kimura 2-Parameter (K2P) distances were calculated using MEGA 5.0. The neighbor-joining (NJ) phylogenetic trees were constructed. RESULTS Six out of the seven candidates were not able to be amplified, while P6 loop of trnL (UAA) intron could be used for identification. The length of the P6 loop of trnL(UAA) intron sequence of Daemonorops draco was 45 bp. The maximum K2P intra-specific genetic distances of Daemonorops draco were much smaller than the minimum inter-specific genetic distances between Daemonorops draco and its adulterants. The NJ tree showed that Draconis Sanguis differed from its adulterants obviously. CONCLUSION The P6 loop of trnL (UAA) intron can be used as DNA barcode for the identification of Draconis Sanguis.     

鸦胆子药材及其混伪品的ITS2序列分子鉴定研究

目的:通过分析鸦胆子及其混伪品的ITS2序列,探究鉴定鸦胆子药材及其混伪品的新方法。方法:采用改良的CTAB法提取鸦胆子及其混伪品的基因组DNA,PCR扩增ITS2片段。对其进行双向测序,应用MEGA6.0软件进行序列分析,计算种内、种间遗传距离,构建邻接树。结果:鸦胆子药材ITS2序列长度为230 bp,种内最大K2P遗传距离为0.018,小于其与混伪品的种间平均K2P遗传距离0.493。NJ树结果表明鸦胆子序列和柔毛鸦胆子序列聚为一支,与其他混伪品可明显区分。鸦胆子与柔毛鸦胆子在35位点处有一变异位点,利用此位点能将鸦胆子与柔毛鸦胆子区别开来。结论:ITS2可作为鸦胆子药材及其混伪品鉴定的DNA条形码序列,为保障临床用药安全提供了新的技术手段。     

基于ITS2序列的朱砂根及其混伪品分子鉴定

目的:应用ITS2序列对朱砂根及其混伪品进行鉴定研究,为中药临床准确用药、市场规范化管理等提供依据和保障。方法:对朱砂根及其混伪品进行DNA提取、PCR扩增ITS2序列、双向测序,对序列进行比对、计算遗传距离。结果:朱砂根ITS2序列长度为217 bp,种内有10个变异位点,有9种单倍型,平均G+C含量为59.1%,除变种红凉伞外,朱砂根与其各混伪品的种间最小遗传距离均大于朱砂根种内最大遗传距离,所以利用ITS2序列可以鉴别朱砂根及其混伪品。结论 :实验结果表明ITS2序列可以鉴别朱砂根及其混伪品,但对于其与变种的关系需进一步研究。     

基于UPLC-Q-TOF-MS/MS技术快速鉴定降香中化学成分

目的:降香为我国传统名贵中药材,然目前市场上降香来源品种复杂,质量差异较大,为进一步明确其药效物质基础,采用超高效液相色谱-串联四极杆飞行时间高分辨率质谱(UPLC-Q-TOF-MS/MS)技术快速分析降香甲醇提取物中的化学成分。方法:采用UPLC RRHD SB-C18色谱柱(3.0 mm×100 mm,1.8μm),以0.1%甲酸水溶液-乙腈为流动相进行梯度洗脱,流速0.3 mL·min^-1,柱温40℃;采用电喷雾离子源,负离子模式采集数据。结果:通过一级精确荷质比和二级碎片信息数据,结合文献资料,质谱裂解规律,Mass bank质谱数据库及对照品的保留时间等,从降香的甲醇提取物中初步鉴定83个化学成分,包括18个黄酮类,31个异黄酮类,10个新黄酮类,9个异黄烷类,7个其他类型黄酮和8个其他类成分。结论:UPLCQ-TOF-MS/MS技术方法可快捷、准确、较全面地鉴定降香甲醇提取物中的化学成分,降香的主要化学成分为异黄酮、黄酮、新黄酮、异黄烷等黄酮类,为降香的药效物质基础研究奠定基础,也为降香药材的质量标准提升提供理论依据和技术支持。     

基于ITS2序列的积雪草及其混伪品的分子鉴定

目的：快速准确的鉴别中药积雪草及其混伪品。方法：采用ITS2序列片段,对其进行PCR扩增、测序,运用Codon CodeAligner、MEGA4．1等软件进行序列拼接和分析,构建积雪草及其混伪品的NJ树。应用Koetschan等建立的ITS2数据库及其网站预测ITS2二级结构。结果：积雪草及其混伪品ITS2序列之间存在明显差异,不同积雪草样品在NJ树上独立聚为一支。结论：运用ITS2序列可以准确鉴定积雪草及其伪品。     

基于ITS2序列鉴定苗药金铁锁及其混伪品

目的：本研究利用ITS2序列对苗药金铁锁及其混伪品进行鉴定研究,从而确保药材质量,保障临床用药安全。方法：提取金铁锁药材及其混伪品基因组DNA,PCR扩增并双向测序获得ITS2序列。所有序列经过CodonCode Aligner V3.7.1拼接后,采用MEGA5.1软件进行序列比对,计算种内和种间Kimura2-Parameter（K2P）遗传距离,并构建系统发育树。采用相似性搜索法、最近距离法、邻接（NJ）树法对ITS2序列鉴定能力进行评估。结果：金铁锁药材的ITS2序列长度为229 bp,其ITS2序列种内最大K2P遗传距离小于与混伪品的最小种间K2P遗传距离;应用相似性搜索法表明ITS2序列能够准确鉴定金铁锁药材及其混伪品;基于ITS2序列构建NJ树,金铁锁及其混伪品均表现出良好单系性,均能明显区分。结论：ITS2序列能够稳定、准确地鉴定金铁锁药材及其混伪品,DNA 条形码技术可为金铁锁药材及其混伪品的鉴定提供新的方法。     

基于ITS2 条形码序列鉴定中药材两头尖及其混伪品

目的：本研究利用ITS2 序列对中药材两头尖及其混伪品进行DNA 条形码鉴定,以确保该药材的质量及临床疗效。方法：采用试剂盒法对36 份样品提取DNA,通过聚合酶链式反应（PCR）扩增其ITS2 序列并双向测序,所得序列经过CodonCode Aligner 拼接后,用软件MEGA5.1 进行数据分析,包括变异位点分析、种内种间距离计算及系统发育树构建。结果：两头尖药材的ITS2 序列长度为216 bp,种内最大K2P 距离为0.014,种间的最小K2P 距离为0.021;NJ 树结果显示两头尖与其混伪品可明显区分,表现出良好的单系性。结论：ITS2 序列作为DNA 条形码能稳定、准确鉴别两头尖药材,为其鉴定提供新的技术手段。     

基于ITS2条形码序列鉴定中药材佩兰及其混伪品

目的:对中药材佩兰及其混伪品进行ITS2条形码鉴定,以确保该药材的质量以及临床疗效。方法:提取佩兰的DNA,利用PCR技术扩增其ITS2序列,双向测序,所得序列经过CodonCodeAligner拼接后,用软件MEGA5进行相关数据分析,计算其种内、种间遗传距离,并构建系统发育树。结果:佩兰药材的ITS2序列长度为218 bp;佩兰种内最大K2P距离为0.009,与混伪品种间最小K2P距离为0.024。ITS2序列的NJ系统聚类树可明显区分佩兰药材及其混伪品,表现出良好的单系性。结论:ITS2条形码序列能够成功鉴定中药材佩兰与其混伪品,为保障临床安全用药提供了新的技术方法。     

蔓荆子药材及其混伪品DNA条形码鉴定研究

本研究应用ITS2序列鉴别蔓荆子及其混伪品,收集药材和基原植物样本共46份,通过提取基因组DNA、PCR扩增和双向测序获得ITS2序列,经CodonCode Aligner V4.2拼接注释获得序列,应用MEGA5.0对序列进行分析比对,计算种内和种间遗传距离（Kimura 2-Parameter,K2P）,构建系统发育NJ树进行鉴定。结果表明,蔓荆子药材基于ITS2序列的种内最大K2P遗传距离均小于其与混伪品的种间最小K2P遗传距离,NJ树显示蔓荆子药材与其混伪品可明显分开,表现出良好的单系性。因此,应用ITS2序列能够准确地鉴定蔓荆子药材及其混伪品。     

1H-NMR技术结合多元统计分析鉴别降香及其伪品

目的：采用核磁共振氢谱(¹H-NMR)技术结合多元统计分析建立降香及其伪品的鉴别方法。方法：获得降香及其伪品的¹H-NMR图谱,建立全成分信息并转化为数据矩阵,检测条件为以含0.03%四甲基硅烷(TMS)的氘代二甲基亚砜(DMSO-d₆)为溶剂,恒温298 K(1 K=-272.15℃),脉冲间隔1.00 s,谱宽12 019.23 Hz,扫描数16次,采样时间1.08 s。对降香及其伪品数据矩阵进行相似度考察和层次聚类分析(HCA),采用正交偏最小二乘法-判别分析(OPLS-DA)对数据矩阵进行分析,找出降香与其伪品之间的差异性成分;建立OPLS-DA类别变量值模型,设定降香类别变量值为1,降香伪品类别变量值为0,阈值±0.3,用于市售降香的真伪鉴别;以OPLS-DA得分图确定市售降香中伪品的种类,并通过薄层色谱法(TLC)进行验证。结果：相似度分析与HCA结果表明,降香与其伪品之间差异明显;OPLS-DA发现降香与其伪品显著性差异成分为反式-橙花叔醇;建立的类别变量值模型可成功对市售降香进行真伪鉴别;OPLS-DA得...