目的 本研究拟通过分析触珠蛋白-CD163-氧应激诱导型血红素加氧酶1(Hp-CD163-HO-1)通路主要基因的交互作用,探索其与2型糖尿病(T2DM)患者冠状动脉粥样硬化性心脏病(冠心病,CAD)发生及病变复杂程度的关联.方法 入选2010年至2015年于阜外心血管病医院行冠状动脉(冠脉)造影检查的T2DM患者共573例,依据造影检查结果分为T2DM+CAD组(n=301)及T2DM-CAD组(n=272);选取T2DM+CAD组中未合并既往事件、经皮冠脉介入治疗、或行冠脉搭桥术的病例作为SYNTAX组(n=131),应用生物信息学筛选通路tagSNPs,以二次PCR及qRT-PCR法进行基因型检测,使用Plink软件及广义多因子降维法统计方法分析tagSNPs及其交互作用在T2DM患者中与CAD发生风险及病变复杂程度的关联.结果 CD163 rs11054072 GA型、白介素4(IL-4)rs3756074 TT型及氧应激诱导型血红素加氧酶1(HMOX1)rs743811 TT型与T2DM患者CAD风险相关;HMOX1 rs743811 TC型及干扰素-γ(IFN-γ)rs2069705 AG型与T2DM合并CAD患者的SYNTAX评分相关.IFNγrs2069705与白介素10(IL-10)rs3790622,白介素6(IL-6)rs1800796、HMOX1 rs743811与IL10 rs1800871分别存在交互作用,并且与T2DM患者冠脉SYNTAX评分相关(P<0.05).结论 Hp-CD163-HO-1通路基因的多态性及交互作用与T2DM患者CAD的发生风险和病变复杂程度有关.

目的：验证并比较“阜华模型2”与其他已发表的国内外华法林剂量预测模型的预测准确性。<br>　　方法：随机入选2014-09至2015-09在阜外心血管病医院心外科住院进行人工瓣膜置换术患者，术后服用华法林药物达到稳定剂量，坚持服用华法林药物治疗至少2个月以上的中国汉族人群。利用Pubmed，Medline，Embase数据库检索2004年后发表的华法林剂量预测模型，采用绝对误差均值（MAE）和预测百分比比较“阜华模型2”及其他已发表模型预测准确性。本研究已在Clinicaltrials.gov试验注册网站进行注册（NCT01855737）。

OBJECTIVETo evaluate the effect of VKORC1, CYP2C9, GGCX, PROC, EPHX1 and CYP4F2 gene polymorphisms on Warfarin maintenance dose variation in Chinese Han Population.METHODSFour hundred eighty-eight patients with prosthetic heart valves, atrial fibrillation or pulmonary thromboembolism and achieved stable Warfarin dose were enrolled. TaqMan probe or direct sequencing were used to genotype Y9VKORC1, CYP2C9, GGCX, EPHX1 and CYP4F2 gene polymorphisms. Demographic characteristics, stable therapeutic dose of Warfarin and concomitant medications were collected for all patients. The effect of VKORC1, CYP2C9, GGCX, PROC, EPHX1 and CYP4F2 gene polymorphisms, demographic characteristics and concomitant medications on Warfarin daily maintenance dose were analyzed with statistical method.RESULTSVKORC1 and CYP2C9 gene polymorphisms could explain more than 50% Warfarin maintenance dose variation in recruited patients, while CYP4F2 gene polymorphisms could only explain 1%. GGCX, PROC and EPHX1 gene polymorphisms had no impact no Warfarin maintenance dose. VKORC1 and CYP2C9 gene polymorphisms have a greater impact on Warfarin maintenance dose compared with demographic characteristics and concomitant medications.CONCLUSIONVKORC1 and CYP2C9 gene polymorphisms have a significant impact on Warfarin maintenance dose in Chinese Han population.

BACKGROUND:Whether two clopidogrel pretreatment strategies prior to elective percutaneous coronary intervention (PCI): a 300 mg loading dose (LD) in clopidogrel naїve patients and a 75 mg maintenance dose (MD) once daily in patients on chronic clopidogrel therapy play the same role in the platelet inhibition in Chinese with different CYP2C19 genotypes remains unknown. We aim to evaluate the impact on platelet inhibition by clopidogrel pretreatment strategy and its interaction effect with CYP2C19 genotype.METHODS:Chinese patients undergoing PCI (n = 840) were assigned to 2×2 groups in the trial according to different clopidogrel pretreatment strategies (470 patients in LD, 370 patients in MD) and CYP2C19 genotypes (494 carriers of any CYP2C19 *2 or *3 loss-of-function allele, 346 non-carriers). The primary outcome was platelet aggregation (PA) as measured by the 10 µmol/L adenosine diphosphate induced light transmission aggregation.RESULTS:Compared with MD group, LD strategy showed a significantly higher PA-((59.22 ± 11.67)% vs. (52.83 ± 12.17)%, P < 0.01), similar PA difference was observed in CYP2C19 loss-of-function carriers compared with non-carriers ((59.41 ± 10.91)% vs. (52.10 ± 12.90)%, P < 0.01). LD patients in either the CYP2C19 loss-of-function allele carrier or non-carrier group showed a significantly higher PA compared with MD group ((61.50 ± 10.61)% vs. (56.84 ± 10.74)%, P < 0.01; (56.06 ± 12.34)% vs. (46.88 ± 11.78)%, P < 0.01, respectively). A quantitative interaction effect was observed between clopidogrel pretreatment strategy and CYP2C19 genotype (P = 0.001).CONCLUSION:The 300 mg LD strategy results in a decreased effect on platelet inhibition compared with the 75 mg MD in Chinese patients receiving clopidogrel prior to PCI, especially in the CYP2C19 2 or 3 loss-of-function allele non-carriers. (ClinicalTrials.gov number NCT01710436)

目的:比较研究BCRP基因多态性对瑞舒伐他汀单剂量和多剂量连续给药后人体药动学的影响。方法:筛选24例健康受试者,按BCRP 421 C>A基因型分组:421CC野生组（n=15）和421CA+AA突变组（n=9）。受试者每日口服规定剂量的瑞舒伐他汀,连续7 d;每日按规定时间点采集受试者血样和尿样;采用液相色谱-串联质谱联用法（HPLC-MS/MS）测定血浆和尿样中的瑞舒伐他汀浓度。结果:单剂量给药后,瑞舒伐他汀在BCRP 421CA+AA突变组的AUC0～t,C max和尿药排泄百分数Percent e xcretion显著高于421CC野生型组（P=0.030,0.015和0.041）,半衰期t1/2和达峰时间T max在两组人群中无差异（P>0.05）。与单剂量结果不同,多剂量连续给药达稳态后,稳态AUC ss,C max、总清除率CL total s s/F和肾清除率CL R s s在野生型组和突变组之间均无显著性差异（P>0.05）,但半衰期t1/2在野生组显著延长[（14.1±3.1）vs（11.8±2.2）h,P=0.035],且野生组的蓄积比Rac也高于突变组（P=0.064）。结论:BCRP 421C>A基因多态性是影响瑞舒伐他汀人体药动学改变的重要因素。

OBJECTIVES:To establish an algorithm to predict the warfarin maintenance dose in Chinese Han population and validate the accuracy of this algorithm.METHODS:A total of 488 Chinese Han patients, hospitalized in Fuwai hospital and had a stable dose of warfarin and a target international normalized ratio (INR) of 1.5 to 3.0, were recruited. Indications for warfarin use included prosthetic heart valve, atrial fibrillation and pulmonary embolism. These patients were divided into derivation group (n = 323) and validation group (n = 165) according to the enrollment time. A warfarin maintenance dose algorithm was established based on genetic information, demographic characteristics and concomitant medications by multiple linear regression analysis in derivation group. In the validation group, we evaluated the accuracy of our algorithm by comparing the predicted dose with the actual dose.RESULTS:Our algorithm included VKORC1-1639G > A, CYP2C9*3 and CYP4F2 genotype, age, Body hight, body weight, amiodarone and digoxin use (R(2) = 0.652, P < 0.001) .In the validation group, the average predicted dose by our algorithm had no statistical difference with the actual dose [(3.51 ± 1.03) mg vs. (3.53 ± 1.41) mg, P = 0.779]. Our algorithm identified 100 out of 165 (60.6%) patients in the validation group, whose predicted dose of warfarin was within 20% of the actual dose, and predicted warfarin dose was underestimated in 17.6% (29/165) patients and overestimated in 21.8% (36/165) patients.CONCLUSION:Our algorithm based on VKORC1, CYP2C9 and CYP4F2 polymorphisms can help to predict the warfarin maintenance dose in Chinese Han Population.

目的:探讨脂滴包被蛋白(perilipin,PLIN)基因多态性与原发性高血压患者美托洛尔治疗后甘油三酯升高的相关性。方法:入选患者97例,服用美托洛尔缓释片100 mg·d-1,持续8周;服药前后分别测量血糖、甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)水平;直接测序法分析PLIN rs2304796 C>T多态性;使用SPSS 15.0统计软件进行数据分析。结果:服用美托洛尔后,患者血浆TG较服药前升高[(2.05±2.42)vs(2.38±2.71)mmol·L-1,P=0.016)]。血糖、TC、HDL-C、LDL-C服药前后无明显变化。PLIN rs2304796 C>T多态性与美托洛尔服药后TG升高相关,其中,CC型携带者升高(0.23±1.02)mmol·L-1,CT型升高(0.26±1.07)mmol·L-1,TT型升高(1.40±3.21)mmol·L-1,P=0.047。结论:perilipin蛋白rs2304796 C>T多态性与美托洛尔服用后TG的升高相关,TT基因型携带者TG水平升高明显。

Objective: To study the effect of cytochrome P-450 4F2 (CYP4F2, rs2108622) gene polymorphisms in patients with warfarin for initial doses in 7 days. <br> Methods: A total of 271 patients treated by warfarin were studied. The CYP4F2 gene polymorphisms were assessed by real-time PCR, the average initial warfarin doses in 7 days and the time of international normalized ratio (INR) ifrst arrived to therapeutic range were recorded. The differences of initial warfarin doses and the time of INR ifrst arrived to therapeutic range among CYP4F2 gene polymorphisms of CC, CT and TT genotypes were analyzed by statistical method. <br> Results: The average initial warfarin doses among CYP4F2 polymorphisms of TT and CT/TT were higher than CC, P<0.05. The times of INR first arrived to therapeutic range in TT genotype was higher than CC genotype, P<0.05. With adjusted gender, age, body height and weight, CYP2C9 and VKORC1 polymorphisms, conditions of disease and medication, the effect of CYP4F2 gene polymorphisms on initial warfarin doses in 7 days were still with the statistic meaning (regression coefifcient=0.21, P=0.003). <br> Conclusion: CYP4F2 polymorphisms inlfuenced the initial warfarin doses in 7 days in relevant patients.

目的：与药效动力学相关的维生素K环氧化物还原酶复合体亚单位1基因（VKORC1）、与药代动力学相关的细胞色素P4502C9基因（cytochrome P4502C9，CYP2C9）的基因多态性是导致华法林用药剂量差异的两个主要基因。细胞色素P4504F2基因（Cytochrome P4504F2，CYP4F2）对华法林剂量影响较小。本研究的目的是确定上述基因在中国汉族人群中个的分布情况。

目的：研究细胞色素P450酶4F2（CYP4F2，rs2108622）基因多态性对中国人群在初始阶段华法林剂量和抗凝效果的影响。

目的：瑞舒伐他汀是一种新型他汀类降脂药，体外细胞试验结果表明， OATP1B1和BCRP/MRP2转运体分别介导该药的肝细胞摄取和胆汁外排。由于他汀类药物是长期连续用药，在多剂量连续给药条件下观察肝胆转运体基因型对药物体内过程的影响更有意义。目前转运体基因型对瑞舒伐他汀多剂量给药体内过程的影响未见报道。因此，本文旨在研究OATP1B1/BCRP/MRP2基因多态性对瑞舒伐他汀单剂量和多剂量给药后人体药代动力学的影响。

目的：评价不同华法林剂量预测模型对于中国汉族人群华法林稳定剂量的预测价值。<br>　　方法：随机选取488例阜外医院门诊或住院的正在服用华法林抗凝的中国汉族患者，抗凝指证包括人工瓣膜置换术后、房颤及肺血栓栓塞症，所有患者服用华法林至少2月以上，且INR均稳定达标，目标INR值在1.5-3.0之间。按患者入选时间分为建模组（323例）和验证组（165例）；对所有入选患者进行VKORC1、CYP2C9、CYP4F2基因多态性检测，记录患者平均每日华法林稳定剂量、人口学信息及合并用药情况等；在建模组患者中，应用多元回归方法，建立治疗窗为INR1.5-3.0中国汉族患者华法林稳定剂量预测模型（阜华模型）；在验证组患者中，通过对比患者的实际剂量与预测剂量的差异，评价阜华模型、国际华法林药物基因组协会（IWPC）推荐的华法林剂量预测模型、Miao及Huang的华法林剂量预测模型的准确性。

Objective To evaluate the effects of CYP2C9 genetic polymorphisms on response to warfarin during initial anticoagulation in Chinese patients with mechanical heart valve replacement.Methods In 254 Chinese valve replacement patients starting warfarin therapy,the CYP2C9 genotypes(CYP2C9*1,*2,and *3) were assessed.The study outcomes included the time to the first International Normalized Ratio(INR) reach the therapeutic range,the total warfarin doses in initial 5 days,and whether INR within the therapeutic range in the day 6.Results Compared with patients with the *1/*1 genotype,patients with the *2 or *3 genotype of CYP2C9 had the same time to the first INR reach the therapeutic range(P=0.933) and no difference in total warfarin doses in initial 5 days(P=0.669).However,the CYP2C9 genotype was a significant predictor of INR within the therapeutic range in the day 6.Conclusion CYP2C9 genetic polymorphisms only affected INR within the therapeutic range in the day 6 in Chinese patients with mechanical heart valve replacement.

Objective To evaluate the microRNAs expression levels in frozen plasma at different frozen time and the stability of microRNAs in total RNA extracted and frozen for 2 weeks.Methods MirVana PARIS Kit was used to isolate total RNA from plasma frozen at-80 ℃ for 2 years,1 years and 6 months and plasma of 10 healthy individuals.The expression levels of 4 microRNAs were detected with TaqMan microRNA assays quantitative real-time PCR.Results There were no significant differences in the levels of the same microRNAs between fresh plasma and plasma frozen for 2 years,1 year or 6 months except U6 in plasma frozen for 6 months(P0.05).There was no significant difference between microRNAs relative expression levels being isolated for 2 weeks RNA and being isolated instantly(P0.05).Conclusion MicroRNAs in plasma frozen at-80 ℃ for 2 years are stable.Total RNA extracted and isolated for 2 weeks can be used for quantitative detection of microRNAs.

Objective: To study the pharmacokinetics and bioequivalence of domestic made clopidogrel in healthy Chinese volunteers. Methods: The test formulation was Taijia,clopidogrel 75 mg tablet,and the reference formulation was Boliwei,clopidogrel 75 mg tablet.A single oral administration dose of 150 mg Taijia and 150 mg Boliwei were given to 23 healthy Chinese male volunteers by an open randomized crossover design.The plasma concentrations of clopidogrel and its main metabolite,clopidogrel acid were examined by liquid chromatography-tandem mass spectrometry to explore the pharmacokinetics and bioequivalence of two formulations. Results: The main pharmacokinetic parameters of plasma clopidogrel in Test group and in Reference group:the value of Cmax(3.07±3.63)and(2.67±2.35)ng/ml;t1/2(6.57±3.18)and(6.96±3.92)h;AUC0-t(4.75±4.68)and(4.60±4.20)ng·h/ml respectively.The main pharmacokinetic parameters of plasma clopidogrel acid in Test group and in Reference group:the value of Cmax(6 724±1 899)and(6 262±1 968)ng/ml;t1/2(8.77±1.20)and(10.11±8.99)h;AUC0-t(20 702±5 579)and(19 817±4 232)ng·h/ml respectively.In test formulation to reference formulation,the relative bioavailability of clopidogrel and clopidogrel acid were(112.8±42.4)% and(107.4±31.6)% respectively. Conclusion: Our work presented that two formulations have bioequivalence by statistic analysis including two one-side t-test and 90% CI calculation.

Identification of secreted proteins of lung cancer could provide new candidates of serum biomarkers for cancer diagnosis or targets for therapeutic intervention. In this study, we developed a novel strategy that combined functional monoclonal antibody library screening technique and mass spectrometry to identify functional secreted proteins. BALB/c mice were immunized with cancer cells isolated from fresh human lung cancer tissues. The monoclonal antibody library containing 1160 mAbs was established with the mouse spleen cells, whose serum had most anti-proliferative effect on lung cancer cells. Monoclonal antibodies were subjected to an immunoreactive and functional screen and monoclonal antibodies that reacted strongly with secreted proteins in condition medium and lung cancer tissues with high inhibotion of cell proliferation were selected. Antigens that recognized by antibodies were obtained by immunoprecipitation and then identified by mass spectrometry. Mac-2-binding protein (Mac-2BP), the antigen of 13H3 antibody, was identified using this approach. Functional studies demonstrated that the 13H3 antibody suppressed lung cancer cell lines ANIP-973 and A549 proliferation in vitro and inhibit ANIP973 xenograft tumors growth in vivo by inducing cell-cycle arrest at G1 phase, with up-regulation of p27 and down-regulation of cyclin D1. Moreover, the serum level of Mac-2BP was significantly higher in lung cancer patients than healthy controls. At a cutoff value of 6 μg/ml, Mac-2BP might be a diagnostic biomarker of lung cancer, especially for SCLC. Mac-2BP concentrations of 6 μg/ml or higher was associated with poor overall survival in univariate analysis, and was an independent predictor in the multivariate COX analysis. Together, these results firstly demonstrated that Mac-2BP can be used as a therapeutic target and potential biomarker for lung cancer. Our strategy is feasible, which may facilitate the identification of novel secreted biomarkers of lung cancer.