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Abstract CYP2C19 gene has multiple single nucleotide polymorphism (SNP), which is the major determinant for clopidogrel treatment responses. Therefore, CYP2C19 SNP detection is essential for predicting clopidogrel efficacy. Currently, there is still no quick and effective method for routine detection of common CYP2C19 SNPs in clinical laboratories, which is critically needed prior to clopidogrel treatment. AllGlo™ based quantitative PCR was used to develop a novel genotyping method for CYP2C19 SNP detection, termed CyPAllGlo. The performance of CyPAllGlo was compared with that of the commonly used fluorescence in situ hybridization (FISH) method, and the data was verified by DNA sequencing. CyPallGlo was used to identify CYP2C19 polymorphisms in 363 patients with coronary heart disease. The univariate analysis was used to access the antiplatelet efficacy of clopidogrel in patients. The associations between CYP2C19 polymorphisms and clopidogrel efficacy were analyzed. Using CyPAllGlo to detect CYP2C19*2 and CYP2C19*3 alleles was highly specific and fast. The detection limit was approximately 0.07 µg/µl and 0.7 µg/µl for CYP2C19*2 and CYP2C19*3, respectively. The consistency between FISH and CyPAllGlo were 98.07% for CYP2C19*2 and 99.17% for CYP2C19*3. DNA sequencing showed that the accuracy of CyPAllGlo was 100%. The analysis time for the whole CyPAllGlo procedure was approximately 60 min. Univariate analysis showed that the anticoagulation efficacy of clopidogrel was related to patient age, CYP2C19 genotype, metabolic phenotype, and LDL level. The logistic regression analysis showed that the genotype of CYP2C19 and metabolic phenotype was the two risk factors for clopidogrel antiplatelet ineffectiveness. This novel CyPAllGlo is a rapid and accurate method for detection of CYP2C19 SNP. The specificity and consistency of CyPAllGlo are comparable with that of widely used DNA sequencing. These findings provide valuable rapid method for predicting clopidogrel efficacy, which can be quickly translated to improve personalized precision medicine for coronary heart disease treatment.

Abstract Hypertrophic cardiomyopathy (HCM), a genetically and clinically heterogeneous cardiomyopathy, is commonly caused by mutations in the MYBPC3 gene or other various sarcomeric genes. HCM patients carrying sarcomeric gene mutations may experience an asymptomatic period at early stage but still possess an escalating risk of developing adverse cardiac events including sudden cardiac death. It is crucial to determine the phenotypic and pathogenic effects of mutations in sarcomeric genes. In this study, a 65‐year‐old male was admitted with a history of chest pain, dyspnoea, and syncope and with a family history of HCM and sudden cardiac death. On admission, electrocardiogram indicated atrial fibrillation and myocardial infarction. Transthoracic echocardiography revealed left ventricular concentric hypertrophy and systolic dysfunction (48%), which were ascertained by cardiovascular magnetic resonance. With late gadolinium‐enhancement imaging, cardiovascular magnetic resonance found myocardial fibrosis on left ventricular wall. The exercise stress echocardiography test showed non‐obstructive myocardial changes. Whole‐exome sequencing analysis identified a MYBPC3 gene heterozygous nonsense variant (c.1522C>T) in the patient and one of his healthy grandnieces (18‐year‐old). The patient was diagnosed with non‐obstructive HCM, heart failure, atrial fibrillation, and so on. Medications, ICD implantation, and catheter ablation were chosen to maintain heart function. Our study provides the clinical evidence regarding the HCM pathogenicity of MYBPC3 c.1522C>T variant and highlights the significance of family genetic testing in the diagnosis and management of HCM.

In plants, nucleotide-binding site and leucine-rich repeat proteins (NLRs) play pivotal roles in effector-triggered immunity (ETI). However, the precise mechanisms underlying NLR-mediated disease resistance remain elusive. Previous studies have demonstrated that the NLR gene pair Pik-H4 confers resistance to rice blast disease by interacting with the transcription factor OsBIHD1, consequently leading to the upregulation of hormone pathways. In the present study, we identified an RNA recognition motif (RRM) protein, OsRRM2, which interacted with Pik1-H4 and Pik2-H4 in vesicles and chloroplasts. OsRRM2 exhibited a modest influence on Pik-H4-mediated rice blast resistance by upregulating resistance genes and genes associated with chloroplast immunity. Moreover, the RNA-binding sequence of OsRRM2 was elucidated using systematic evolution of ligands by exponential enrichment. Transcriptome analysis further indicated that OsRRM2 promoted RNA editing of the chloroplastic gene ndhB. Collectively, our findings uncovered a chloroplastic RRM protein that facilitated the translocation of the NLR gene pair and modulated chloroplast immunity, thereby bridging the gap between ETI and chloroplast immunity.

【Objective】IQD gene family proteins are plant specific calmodulin-binding target proteins, which play an important role in plant defense and development regulation. The interaction between Pik-H4 and OsIQD1 was found in the yeast double hybrid screening experiment in laboratory. In order to reveal the specific biological functions of OsIQD1 in rice blast resistance, OsIQD1 gene was edited by using CRISPR/Cas9 editing technique.【Method】OsIQD1 was obtained by sequence alignment. Two 20 bp editing targets were designed in exon 1 and exon 5 of OsIQD1 (DUF4005 domain), and the nucleotide fragments of the two targets were cloned into pRGEB32 vector to obtain pRGEB32-OsIQD1 -gRNA vector. The rice Pik-H4 NIL callus was transformed by agrobacterium-mediated method. Transgenic positive plants were obtained by regeneration culture and hygromycin resistance screening. PCR and sequencing were performed on the target region sequences of transgenic plants of T0 generation to analyze the mutation types of osiqd1.【Result】The transgenic regenerated plants were identified by PCR with hygromycin resistance gene HPT Ⅱ, and 30 positive lines were obtained. Sequencing analysis of the sequences near the target site of T0 generation plants showed that OsIQD1 gene was successfully edited, and the editing efficiency of the two sites was 21.67% and 26.67%, respectively. There was one homozygous mutant line in target 2 region.【Conclusion】The study results provide genetic material for further studies on the mechanism of osiqd1 mutation involved in calcium ion/calmodulin signaling pathway, and OsIQD1 is preliminarily presumed to be involved in the basic immune response of plants.

The intricate regulatory process governing rice immunity against the blast fungus Magnaporthe oryzae remains a central focus in plant–pathogen interactions. In this study, we investigated the important role of Osa-miR11117, an intergenic microRNA, in regulating rice defense mechanisms. Stem-loop qRT-PCR analysis showed that Osa-miR11117 is responsive to M. oryzae infection, and overexpression of Osa-miR11117 compromises blast resistance. Green fluorescent protein (GFP)-based reporter assay indicated OsPAO4 is one direct target of Osa-miR11117. Furthermore, qRT-PCR analysis showed that OsPAO4 reacts to M. oryzae infection and polyamine (PA) treatment. In addition, OsPAO4 regulates rice resistance to M. oryzae through the regulation of PA accumulation and the expression of the ethylene (ETH) signaling genes. Taken together, these results suggest that Osa-miR11117 is targeting OsPAO4 to regulate blast resistance by adjusting PA metabolism and ETH signaling pathways.

Background and aimsThe number of hypertensive population rises year by year recently, and their age becomes more youthful. For a long time, hypertension has long been regarded as a multi-factorial disease. In addition to smoking, genetics, diet and other factors, helicobacter pylori (H. pylori) had been regarded as a potential risk factor for hypertension in recent years. However, most studies had certain limitations and their results were inconsistent. Thus, it is necessary for us to assess the impact of H. pylori on hypertension through meta-analysis.MethodsWe searched all published relevant literature through multiple databases by July 23, 2021. Pooled results were calculated under the random effect model. Heterogeneity was evaluated by the Q statistic and the I2 statistic. The risk of bias was evaluated via ROBINS-I tool. Publication bias was evaluated by the Egger test and Begg funnel plot.Results6 eligible studies involving 11317 hypertensive patients and 12765 controls were selected from 20767 retrieval records. Our research confirmed that H. pylori significantly increased the probability of suffering from hypertension in the random effect model (OR:1.34, 95% CI:1.10-1.63, P = 0.002, I2 = 74%). The same results were also found in both Asian population and developing country (OR:1.28, 95%CI:1.05-1.55, P = 0.003, I2 = 78.5%).ConclusionsOur results confirmed that H. pylori was a vital risk factor for hypertension. H. pylori-infected people were 13.4% higher risk for hypertension than uninfected individuals. In addition, it will be a new method to prevent and treat hypertension by eradicating H. pylori.Trial registrationThe registration number for systematic review in PROSPERO CRD42021279677.

Objective To investigate the association between DNA methylation and the stable warfarin dose through genome-wide DNA methylation analysis and pyrosequencing assay. Method This study included 161 patients and genome-wide DNA methylation analysis was used to screen potential warfarin dose-associated CpGs through Illumina Infinium HumanMethylation 450 K BeadChip; then, the pyrosequencing assay was used to further validate the association between the stable warfarin dose and alterations in the methylation of the screened CpGs. GenomeStudio Software and R were used to analyze the differentially methylated CpGs. Results The methylation levels of CpGs surrounding the xenobiotic response element (XRE) within the CYP1A1 promoter, differed significantly between the different dose groups (P 0, P

Background We used bioinformatic analysis and quantitative reverse transcription polymerase chain reaction (RT-qPCR) assays to investigate the association between plasma microRNAs (miRNAs) and stable warfarin dosage in a Chinese Han population. Methods Bioinformatics analysis was used to screen out potential warfarin dose-associated miRNAs. Three plasma miRNAs were validated in 99 samples by RT-qPCR. Kruskal–Wallis test and multivariate logistic regression were used to compare differences in plasma miRNAs expression levels between three warfarin dosage groups. Results There were significant between-group differences among the three dose groups for hsa-miR-133b expression (p = 0.005), but we observed an “n-shaped” dose-dependent curve rather than a linear relationship. Expression levels of hsa-miR-24-3p (p = 0.475) and hsa-miR-1276 (p = 0.558) were not significantly different in the multivariate logistic regression. Conclusion miRNAs have received extensive attention as ideal biomarkers and possible therapeutic targets for various diseases. However, they are not yet widely used in precision medicine. Our results indicate that hsa-miR-133b may be a possible reference factor for the warfarin dosage algorithm. These findings emphasize the importance of a comprehensive evaluation of complex relationships in warfarin dose prediction models and provide new avenues for future pharmacogenomics studies.

Bacterial leaf streak (BLS), caused by Xanthomonas oryzae pv. oryzicola (Xoc), is a bacterial disease affecting rice production in Asia and Africa, whose severity is expected to increase with climate change. Identification of new quantitative-trait loci (QTL) or resistance genes for BLS resistance is essential for developing resistant rice. A genome-wide association study to identify QTL associated with BLS resistance was conducted using phenotypic and genotypic data from 429 rice accessions. Of 47 QTL identified, 45 were novel and two co-localized with previously reported QTL or genes conferring BLS resistance. qBLS6.2 on chromosome 6 explained the greatest phenotypic variation. Combined analysis of differential expression and annotations of predicted genes near qBLS6.2 based on haplotype and disease phenotype identified OsBLS6.2 (LOC_Os06g02960) as a candidate gene for qBLS6.2. OsBLS6.2 knockout plants showed higher resistance to Xoc than wild-type plants. Many other candidate genes for resistance to Xoc were identified. [ABSTRACT FROM AUTHOR]