目的 建立一种可同时、快速检测2型糖尿病患者降糖药相关药物基因多态性的液相芯片检测系统.方法 根据7个目的基因的rs号,在Genbank中查找其靶位点附近碱基序列(目的基因包括磺脲类受体1,转录因子7类似物2,胰岛素受体底物1,过氧化物酶体增殖物激活受体γ,有机阳离子转运蛋白与多药和有毒化合物排出家族,有机阴离子转运蛋白家族成员1B1等),以PrimerPlex软件设计等位基因特异性引物,Primer6.0软件设计含检测位点的PCR引物,通过多重PCR扩增,等位基因特异性引物延伸(ASPE),MagPlex～Tag微球杂交,液相芯片系统Luminex 200检测荧光信号,确定基因型,优化反应体系并以5份代表性标本进行方法学评价.收集2019年1月～12月东莞市厚街医院新诊2型糖尿病患者血液样本115例,采用建立的系统检测上述7个靶位点,并随机选取25例与测序结果比较.结果 7个目标基因经PCR扩增,产物电泳成像后清楚可见7条目标条带,无非特异性条带;经优化ASPE杂交条件,选择退火温度55℃,Biotion～dCTP浓度与dCTP浓度的比值为3:1,37℃孵育45 min时检测效果最佳;临床样本检测结果显示:纯合子荧光强度中位值(median fluorescence intensity,MFI)比率均>0.9或<0.1,杂合子MFI比率在0.4～0.6之间.纯合子MFI比率批内批间CV在0.9％～3.3％和2.1％～4.6％,而杂合子则在2.9％～7.3％和5.2％～11.2％;DNA最低检测限为0.75ng;25例样本的检测结果与测序结果完全一致,准确度100％.结论 该研究成功建立了一种新的液相芯片检测系统,高效便捷,能同时检测7个目的基因型,满足临床的需求.

Objective To establish a method for simultaneous and rapid detecting of the polymorphisms in Cytochrome P4502C9 (CYP2C9), CYP2C19, CYP4F2, Vitamin K epoxide reductase (VKORC1) and ATP-binding cassette subfamily B member1 (ABCB1) gene, which were associated with warfarin and clopidogrel, based on liquid phase chip technology. Methods Method establishment. The eight gene sequences near targeted sites related to warfarin and clopidogrel were found in Genbank, and the specific primers and probes were designed. Through multiple PCR amplification, followed by allele specific primer extension (ASPE), and MagPlex-Tag microspheres hybridization, the suspension array Luminex 200 system step-by-step, the genotypes were determined by fluorescence signal. The reaction system was optimized and its methodological evaluation was performed. 260 patients with antithrombotic therapy from Dongguan houjie hospital were recruited in this study form June 2017 to December 2018. The eight genotypes of the 260 patients were detected by the established method, and the results were compared with the sequencing results. Results The results of 260 samples showed that allelic median fluorescence intensity (MFI) ratios of homozygotes (mutant/wild-type) were all greater than 0.9 or less than 0.1, and all the allelic MFI ratios of heterozygotes were between 0.3 and 0.6. The within run and between run coefficients of variance for allelic MFI ratios were lower than 6.4％and 10.9％, respectively. The minimum DNA template requirements was 0.75ng. The genotypes of 260 patients determined by the established method were completely concordant with the sequencing results. Conclusion A method was established successfully for rapid detecting the genotypes which associated with warfarin and clopidogrel based on liquid phase chip technology.

目的:通过检测心肺复苏(CRP)患者血清中S100A8/A9和高迁移率族蛋白B1(HMGB1)水平,探讨S100A8/A9和HMGB1在心肺复苏后脑损伤中的表达与预后的关系.方法:选取心跳骤停(CA)/CRP成功后仍需进行脑复苏患者50例,根据格拉斯哥-匹兹堡脑功能表现分级(CPC)评分标准,将患者分为预后差组(CPC3～5级)和预后良好组(CPC1～2级),分别在入院时、CRP成功时、CRP24 h、CRP48 h抽取患者静脉血检测S100A8/A9和HMGB1水平,取50例正常健康体检人群的血清为对照组.结果:预后差组和预后良好组患者血清中S100A8/A9、HMGB1的表达水平均高于对照组(P<0.05).预后差组S100A8/A9、HMGB1表达水平高于预后良好组(P<0.05).结论:S100A8/A9和HMGB1可以作为血清标志物应用于CRP后缺血再灌注中脑损伤程度及预后判断.

Objective To perform a meta-analysis on the associations of interleukin (IL)-6-174G ＞ C (rs1800795) and-634C ＞ G (rs1800796) polymorphisms with type 2 diabetic nephropathy (DN).Methods The data on the studies about the associations of IL-6-174G ＞ C and-634C ＞ G polymorphisms with type 2 DN were collected from Pubmed,Embace,CNKI,Wan Fang and VIP database during their inception and April 2017.The statistical analysis was performed with STATA 14.0 and Review manager 5.3 softwares.The heterogeneity in the eligible studies was assessed by Q-statistic and I2 statistic.When the significant heterogeneity was found,the random effect model was used for meta-analysis,otherwise,the fixed effect model was used.The publication bias was evaluated with funnel and Begger graphs.The pooled odds ratios (OR) and corresponding 95％ confidence intervals (95％ CI) were calculated for evaluating the associations of IL-6-174G ＞ C and-634C ＞ G polymorphisms with type 2 DN.In addition,the sub-group analysis was performed according to the regions of subjects.Results A total of 11 studies were enrolled,The studies on the association of IL-6-174G ＞ C polymorphism with type 2 DN included 1 688 subjects,while those on the association of IL-6-634C ＞ G with type 2 DN included 2 180 subjects.In the association analysis of IL-6-174G ＞ C polymorphism with type 2 DN of Asian population,the significant relationship was detected in an allelic genetic model (OR =0.461,95％ CI:0.274-0.777,P ＜ 0.01),a homozygote model (OR =0.126,95％ CI:0.022-0.734,P =0.021),a recessive genetic model (OR =0.146,95％ CI:0.026-0.827,P =0.030) and a dominant genetic model (OR =0.504,95 ％ CI:0.273-0.930,P =0.028),but not in a heterozygote model (OR =0.606,95 ％ CI:0.321-1.143,P =0.122).There was no significant relationship between IL-6-174G ＞ C polymorphism and type 2 DN in European population.In the association analysis of IL-6-634C ＞ G polymorphism with type 2 DN of Asian population,the significant relationship was found in an allelic genetic model (OR =1.467,95％ CI:1.238-1.737,P ＜0.01),a homozygote model (OR =2.793,95％ CI:1.844-4.230,P =0.021),a recessive genetic model (OR =2.296,95 ％ CI:1.586-3.323,P ＜ 0.01) and a dominant genetic model (OR =1.377,95％CI:1.109-1.711,P ＜ 0.01),but not in a heterozygote model (OR =1.733,95％ CI:0.932-1.476,P =0.174).There was no significant relationship between IL-6-634C ＞ G polymorphism and type 2 DN in European population.Conclusion In Asian population,IL-6-174CC genotype may prevent the progression of type 2 DN,however,IL-6-634GG genotype may promote the development of type 2 DN.But in European population,there is no relationship between IL-6-174G ＞ C and-634C ＞ G polymorphisms and type 2 DN.

In recent years, with the development of precision medicine, the further researches of drugs targeted genomics provided the important basic and theory for individualized clinic treatment. We could improve the therapeutic effect through detecting related drug gene polymorphisms of patients and understand the genotype of drug sensitivity, then determining the types and dosages. However, we urgently need the high efficiency, low cost, high flux detection method in clinical practices. Fortunately, Liquid chip technology as a high-throughput detection technology of new generation, and can meet the above requirements. Now, Liquid chip technology was widely used in genetic polymorphism detection,and provide a new platform for high-throughput gene detection. This paper reviewed the principle of liquid chip technology, technical analysis model of Luminex xMAP detects SNPs and the application prospects of liquid chip technology in the rational use of drugs.