OBJECTIVETo investigate the association of the polymorphisms of cytochrome P450 2C9 (CYP2C9) exon 4 608T/G, 561A/C, 537A/C and 527A/C, and -65G/C with warfarin sensitivity.

METHODSA total of 102 patients under warfarin anticoagulant therapy were selected. During follow-up, warfarin dosage and associated Prothrombin Time-International Normalized Ratio (P-INR) values were recorded. Simultaneous monitoring of incidence of bleeding and thrombosis adverse effect was recommended. Genetic polymorphisms of the above mentioned loci were identified by polymerase chain reaction and DNA sequencing.

RESULTSThe average age of the 102 patients was (62.1+/-10.5) years. The body mass index (BMI) was (24.7+/-3.8) kg/m2. Mean daily warfarin requirement was from 1.250 to 5.077 mg/day when therapeutic PT-INR (1.5-2.5) was maintained. DNA sequencing showed no polymorphisms of 608T/G, 561A/C, 537A/C, 527A/C in CYP2C9 exon 4. Warfarin daily dosage in CYP2C9 exon 4 -65C carriers was 3.106+/-0.619 mg/d, while it was (2.555+/-0.708) mg/d in individuals with wild-type -65G (P=0.020). Receiver operating characteristic (ROC) analysis showed that warfarin daily dosage of more than 2.5 mg/d can be used to predict the CYP2C9 exon 4 -65GC genotype (AUC: 0.770, P=0.005, 95%CI:0.626-0.915). Logistic regression indicated that BMI was an independent factor of bleeding during anti-coagulation therapy (OR=0.794, 95%CI: 0.651-0.970, P=0.024).

CONCLUSIONThe Chinese population are, generally, warfarin-sensitive. Exon 4 of the CYP2C9 gene is highly conserved in this population. The warfarin maintenance dosage in CYP2C9 exon 4 -65CG carriers was significantly higher than those with wild-type -65GG. The clinical significance needs further investigation with more large-scale, multi-center trials.

Adult ; Alleles ; Anticoagulants ; pharmacology ; therapeutic use ; Aryl Hydrocarbon Hydroxylases ; genetics ; Cytochrome P-450 CYP2C9 ; Exons ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Point Mutation ; Polymorphism, Genetic ; Thrombosis ; drug therapy ; Warfarin ; pharmacology ; therapeutic use

BACKGROUNDAspirin is widely used in the secondary prevention of coronary artery diseases, including myocardial infarction, stroke, and vascular related deaths. However, the antiplatelet effect of aspirin appears to be variable and aspirin resistance (AR) is currently still controversial for Chinese patients. The aim of this study was to describe the prevalence of AR, and identify possible risk factors associated with a lack of response to aspirin treatments in patients with unstable coronary artery disease.

METHODSPlatelet function tests with arachidonic acid (ARA) and urinary 11-dehydro-thromboxane B2 (11-DH-TXB2) concentrations were performed in 262 patients with unstable coronary artery disease who had not been taking aspirin before admission. ARA induced platelet aggregation and 11-DH-TXB2 were detected to evaluate the functional and biochemical responses to aspirin before and on days 1, 4, and 10 after aspirin administration. Six-month follow-up was completed in patients who developed AR to evaluate the effect of aspirin in a long-term treatment. GP1Bα (C1018T), Pl (A1/A2), P2Y1 (A1622G), TBXA2R (T924C) were also detected to evaluate the influence of genetic variant on aspirin responsiveness.

RESULTSA total of 8.8% of patients were indentified as AR at the first day after aspirin treatment. The level of urine 11-DH-TXB2 in the AR group was higher compared to non-AR group (P < 0.05). There was no relationship between ARA induced platelet aggregation and urinary 11-DH-TXB2 levels (r = 0.038, P = 0.412). The results of DNA sequencing showed that TBXA2R-924TT homozygotes had a significantly high rate of AR. Logistic regression demonstrated that diabetes was an independent risk factor of AR.

CONCLUSIONSIn the beginning period of administration, aspirin was not a sufficient factor that inhibits platelet aggregation. TBXA2R-924T allele was involved in AR. Diabetes was an independent risk factor of AR.

Aged ; Aged, 80 and over ; Arachidonic Acid ; pharmacology ; Aspirin ; therapeutic use ; Coronary Artery Disease ; drug therapy ; genetics ; Diabetes Mellitus, Type 2 ; Female ; Genotype ; Humans ; Male ; Membrane Glycoproteins ; genetics ; Middle Aged ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; therapeutic use ; Platelet Function Tests ; Platelet Glycoprotein GPIb-IX Complex ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Receptors, Purinergic P2Y1 ; genetics ; Receptors, Thromboxane A2, Prostaglandin H2 ; genetics ; Thromboxane B2 ; analogs & derivatives ; urine

Objective To evaluate the changes of the right atrium,right ventriculum,left atrium and left ventriculum after transcatheter closure of atrial septal defect(ASD) during a short to mid-term follow-up.Methods The right ventricular end-diastolic anterior-posterior diameter(RVEDD),right atrial long diameter(RADl),right atrial transverse diameter(RADt),left ventricular end-diastolic ante-posterior diameter(LVEDD),left ventricular end-diastolic volume(LVEDV) and left atrial anterior-posterior diameter(LAD) in 36 patients with secundum ASD were measured before ASD closure,after 3 days,3 months and 6 months of ASD closure with transthoracic echocardiography(TTE).Results RVEDD,RADl and RADt were significantly decreased,while LVEDD,LVEDV and LAD significantly increased 3 days after ASD closure.During 3 months follow-up,RVEDD,RADl and RADt continuously became smaller;LVEDD,LVEDV and LAD continuously became larger.At 6 months,RVEDD was significantly smaller and LVEDD,LVEDV were significantly larger than those at 3 months.No remarkable difference of RADl,RADt and LAD was found between 6 months and 3months follow-up.Conclusion Transcatheter closure of ASD not only decreases the preload of right heart and causes right atrium and right ventriculum become smaller,but also improves the geometry of left heart and causes the narrowed left atrium and left ventriculum gradually return to almost normal status.

Objective To study the pathogenesis of nasopharyngeal carcinoma and identify potential biomarkers or therapeutic targets. Methods Microarray data (GSE12452 and GSE13597) were downloaded from Gene Expression Omnibus. Processing of original microarray data and screening of differentially expressed genes were performed through bioinformatics analysis. Then, GO and KEGG pathway enrichment analysis was performed for these genes using DAVID database. Real time-PCR and Western blot assay were used to detect the expression levels of the identified genes. Results A total of 260 overlap DEGs were obtained including 16 GO entries and 4 signal pathways. Eighteen potential therapeutic targets that relative to cell cycle were identified by gene enrichment analysis. Expression levels of 12 selected genes were confirmed by real-time PCR. Finally, 4 selected genes were confirmed by Western blot assay. Conclusion By bioinformatics analysis of two sets of microarray data and molecular biology research, four genes were found including CDC6, CDK1,MCM2 and CCNB1, which might be potential key genes that can be developed for therapy targets of NPC in the future.

Objective To assess the antibody test for the diagnosis performance of heparin induced thrombocytopenia(HIT).Methods 52 plasma samples of patients with HIT,126 plasma samples of heparin treated patients without HIT and 50 plasma samples of healthy individuals were collected from 2014 September to 2016 November.According to thrombosis,the patients were further divided into two groups:isolated HIT group without thrombosis (30 cases) and heparin induced thrombocytopenia with thrombosis (HITY) group (22 cases).The whole HIT antibody in plasma was assayed by using ACL-TOP 700 coagulation analyzer and reagent (HemosIL HIT-AbPF4-H).The IgG-specific HIT antibody in plasma was assayed by using ACL AcuStar chemiluminescent analyzer and reagent (HemosIL AcuStar HIT-IgGPF4-H).Results The levels of whole antibody and IgG-specific antibody in the patients of heparin control group was higher than those in healthy control (U value was 1 644.0 and 1 911.0,respectively,P ＜ 0.01).The levels of two HIT antibodies in HIT patients group were higher than those in the patients of heparin control group (U value was 550.0 and 4.7,respectively,P ＜ 0.01).ROC curve showed that the sensitivities of both whole antibody and IgG-specific antibody were 100％,and up-regulating the cut-off value could improve the specificity of both tests.The positive incidence of the whole antibody was 27.8％ in the heparin control group and 100％ in HIT patients group while the cut-off value was 1.50 U/mL.The positive incidence of IgG-specific antibody was 0 in the heparin control group and 100％ in the HIT patients group while the cut-off value was 1.51 U/mL.While the cut-off value of IgG-specific antibody was 2.32 U/mL,the diagnosis sensitivity of thrombosis assessment was 90.9％ and the specificity was 80.0％.In case the cut-off value exceeded 2.32 U/mL,the accumulating risk of HIT increased significantly in HIT patients within 15 days (Log-rank x2 =56.577,P ＜ 0.01).Conclusion The whole antibody and IgG-specific antibody could contribute to excluding diagnosis,diagnosis or risk assessment for the suspected HIT patients.

Objective To assess stroke risk in the patients with new-onset atrial fibrillation (AF) after elective percutaneous coronary intervention (PCI) by von Willebrand factor antigen (vWF∶Ag).Methods A total of 127 patients with new-onset atrial fibrillation after PCI (new-onset AF after PCI group),50 new inpatients with acute coronary syndrome without undergoing PCI (ACS control),50 patients without AF after PCI (PCI control) and 50 healthy individuals (healthy control) were selected in Tianjin Medical University General Hospital from 2016 February to 2017 February for retrospective cohort study.To assay the plasma vWF∶Ag for all objects,receiver operating characteristic curve (ROC) was used to analyze the predicting performance of vWF∶Ag with stroke risk in the patients with new-onset AF.x2 test was used to analyze the relevance between vWF∶Ag and clinical pathological factors in the patients with new-onset AF.Kaplan-Meier curve was used to implement survival analysis.Results The results of plasma vWF∶Ag were as follows:104.5％ (88.0％,133.7％) for the control group,161.7％ (120.7％,270.5％) for ACS control group,208％ (125.2％,360.7％) for PCI control group and 284.9％ (172.4％,494.2％) for new-onset AF after PCI group,respectively.The level of plasma vWF∶Ag of PCI control was higher than that of ACS control (U =526.0,P ＜0.01) and the level in new-onset AF after PCI group was higher than that of PCI control (U =824.0,P ＜ 0.01).The area under the curve was 0.882 (95％ CI:0.811-0.854) for plasma vWF∶Ag level predicting stroke risk with new-onset AF.When the cut-off value of vWF∶Ag was defined as 312.0％,the sensitivity was 94.4％ and specificity was 60.6％.Compared with the patients whose vWF∶Ag was less than 312.0％,the cumulative probability increased significantly (Log-rankx2 =44.308,P ＜ 0.01) during 90 days of follow-up period in the patients with new-onset AF after PCI whose vWF∶Ag was over 312.0％.There were relevances between the vWF∶Ag level and chronic heart failure/left ventricular dysfunction,hypertension,age,stroke/transient ischemic attack (TIA)/history of thromboembolism and vascular disease (P ＜ 0.01).The level of plasma vWF∶Ag and risk of stroke event in the patients with dual antithrombotic therapy were higher than those of the patients with triple antithrombotic therapy (U =1 075.5,P ＜ 0.01;x2 =10.45,P ＜ 0.01).Conclusion The level of plasma vWF ∶ Ag could reflect the damage condition of vascular endothelial cells,stroke risk and the efficacy of anticoagulant therapy in the patients with new-onset atrial fibrillation after PCI.

Objective: To explore the protective effect of Yishen Tongluo Recipe (YTR) against aberrant sperm DNA methylation in male rats exposed to benzo(a)pyrene (BaP). METHODS: Thirty male SD rats of the SPF grade were randomly divided into three groups of equal number: solvent control, BaP exposure and YTR intervention. The animals of the solvent control group were injected intraperitoneally with 0.5% DMSO while those of the other two groups with BaP at 0.1 mg/kg/d, all for 60 days, and at 31 days of BaP exposure, those of the YTR group were treated intragastrically with YTR for 30 days. Then, the left epididymides were harvested from all the rats and sperm suspensions collected and centrifuged for extraction of sperm DNA. The methylated DNA immunoprecipitation sequencing (MeDIP-seq) technique was used to detect the whole-genome DNA methylation in different groups. RESULTS: Exposure to BaP induced the up-regulation of 828 genes encoding mRNA in the sperm DNA, while YTR intervention produced a significant protective effect on the transforming growth factor β3 (TGF-β3), cystic fibrosis transmembrane conductance regulator (CFTR) and recombination activating gene 1 (RAG1), and down-regulated the expressions of 3 227 genes. BaP exposure also caused the up-regulation of 783 genes encoding lncRNA in the sperm DNA, and YTR treatment exhibited an evident protective effect on 62 of the up-regulated genes, induced the down-regulation of 3 378 genes, and showed a protective effect on 56 of the down-regulated genes. CONCLUSIONS YTR has a protective effect against aberrant sperm DNA methylation in male rats exposed to BaP, which may be associated with lncRNA.