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Objective: The purpose of this study was to investigate the effects of CYP2C19 gene mutations on clopidogrel antiplatelet activity in the patients with coronary heart disease treated by percutaneous coronary intervention. Methods: Patients with coronary heart disease, who hospitalized in the Second Affiliated Hospital of Nanchang University from March 2011 to June 2019, and healthy individuals with matching genetic background, gender, and age as controls were included in this study. Basic clinical data were analyzed and blood samples of all research subjects were obtained for extraction of DNA, and Sanger first-generation sequencing method was used to detect CYP2C19 gene mutation from full exon and exon and intron junction. CYP2C19 gene variations in patients with coronary heart disease were compared with the 1000 Genomes Browse database and the sequencing results of healthy controls to determine whether the gene variation was a genetic mutation or a genetic polymorphism. After that, PolyPhen-2 prediction software was used to analyze the harmfulness of gene mutations to predict the effect of mutations on protein function. The same dose of CYP2C19 wild-type plasmid and the CYP2C19 gene mutant plasmids were transfected into human normal liver cells HL-7702. After transfection of 24 h, the expression of CYP2C19 protease in each group was detected. The liver S9 protein was incubated with clopidogrel, acted on platelets to detect the platelet aggregation rate and the activity of human vasodilator-activated phosphoprotein (VASP). Results: A total of 1 493 patients with coronary heart disease (59.36%) were enrolled, the average age was (64.5±10.4) years old, of which 1 129 were male (75.62%). Meanwhile, 1 022 healthy physical examination volunteers (40.64%) were enrolled, and the average age was (64.1±11.0) years old, of which 778 were male (76.13%). A total of 5 gene mutations of CYP2C19 gene were identified in 12 patients (0.80%), namely, 4 known mutations T130K (1 case), M136K (6 cases), N277K (3 cases), V472I (1 case) and one new mutation G27V (1 case), no corresponding gene mutation was found in healthy controls. It was found that T130K and M136K were probably damaging, G27V was possibly damaging, and N277K and V472I were benign mutations. In vitro, we demonstrated that the platelet aggregation rate of the M136K gene mutation group was 24.83% lower than that of the wild type (59.58% vs. 34.75%; P<0.05), and the phosphorylated VASP level was 23.0% higher than that of the wild type (1.0 vs. 1.23; P<0.05). However, the platelet aggregation rate and phosphorylated VASP level were similar between of G27V, T130K, N277K, V472I gene mutation groups and wild type group (P>0.05). Conclusions: In this study, 5 gene mutations are defined in patients with coronary heart disease, namely G27V, T130K, M136K, N277K, V472I. In vitro functional studies show that CYP2C19 gene mutation M136K, as a gain-of-function gene mutation, can enhance the activation of CYP2C19 enzyme on clopidogrel, thereby inhibiting the platelet aggregation rate.
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Objective: The research is to provide the basis for effective protection and wise utilization of the plants resources inTea Mountain National Ecological Demonstration Area of Shandong Province. Methods: The general investigation on plant species inTea Mountain was conducted. The research scope covered plant species, their distribution in wild, community structure and flora.Results: A total of 352 higher plants, belonging to 250 genera and 90 families, were recorded. Four species were listed as nationalsecond-grade protected plants. Regional vegetation is Pinus thunbergii plantation and Secondary forest. The main forest vegetation isPinus thunbergii community. Pantropic types and N. Tem types are dominant types. The latitude and "the islands-effect" should be themajor-effect factor of this pattern. Because of intense artificial active disturbance, primary vegetation types in this region have sufferedserious destruction, especially the rare and precious plants that are at high risk of dying out. Conclusions: This regional communitystructure is simple. The shrub layer and the liana layer are undeveloped. The syngenesis time is short.
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Objective: The research is to provide the basis for effective protection and wise utilization of the plants resources inTea Mountain National Ecological Demonstration Area of Shandong Province. Methods: The general investigation on plant species inTea Mountain was conducted. The research scope covered plant species, their distribution in wild, community structure and flora.Results: A total of 352 higher plants, belonging to 250 genera and 90 families, were recorded. Four species were listed as nationalsecond-grade protected plants. Regional vegetation is Pinus thunbergii plantation and Secondary forest. The main forest vegetation isPinus thunbergii community. Pantropic types and N. Tem types are dominant types. The latitude and "the islands-effect" should be themajor-effect factor of this pattern. Because of intense artificial active disturbance, primary vegetation types in this region have sufferedserious destruction, especially the rare and precious plants that are at high risk of dying out. Conclusions: This regional communitystructure is simple. The shrub layer and the liana layer are undeveloped. The syngenesis time is short.