Targeted metabolomics reveals serum changes of amino acids in mild to moderate ischemic stroke and stroke mimics.

Background: The pathophysiological processes linked to an acute ischemic stroke (IS) can be reflected in the circulating metabolome. Amino acids (AAs) have been demonstrated to be one of the most significant metabolites that can undergo significant alteration after a stroke. Methods: We sought to identify the potential biomarkers for the early detection of IS using an extensive targeted technique for reliable quantification of 27 different AAs based on ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). A cohort with 216 participants was enrolled, including 70 mild to moderate ischemic stroke patients (National Institutes of Health Stroke Scale < 15, MB group), 76 stroke mimics (MM group) and 70 healthy controls (NC group). Results: It was found that upon comparing MB and MM to control patients, AAs shifts were detected via partial least squares discrimination analysis (PLS-DA) and pathway analysis. Interestingly, MB and MM exhibited similar AAs pattern. Moreover, ornithine, asparagine, valine, citrulline, and cysteine were identified for inclusion in a biomarker panel for early-stage stroke detection based upon an AUC of 0.968 (95% CI 0.924-0.998). Levels of ornithine were positively associated with infract volume, 3 months mRS score, and National Institutes of Health Stroke Scale (NIHSS) score in MB. In addition, a metabolites biomarker panel, including ornithine, taurine, phenylalanine, citrulline, cysteine, yielded an AUC of 0.99 (95% CI 0.966-1) which can be employed to effectively discriminate MM patients from control. Conclusion: Overall, alternations in serum AAs are characteristic metabolic features of MB and MM. AAs could serve as promising biomarkers for the early diagnosis of MB patients since mild to moderate IS patients were enrolled in the study. The metabolism of AAs can be considered as a key indicator for both the prevention and treatment of IS.

Molecular mechanism of seed dormancy release induced by fluridone compared with cod stratification in Notopterygium incisum.

BACKGROUND: Notopterygium incisum is an important Chinese medicinal plant. Its mature seeds have underdeveloped embryos and are physiological dormant. We found the seeds with full developed embryos can germinate after treated by fluridone (FL), an inhibitor of abscisic acid (ABA). In order to understand the molecular mechanisms underlying seed dormancy release by FL, we compared the transcriptomic changes in dormancy release induced by two different methods, FL and cold stratification (CS) in N. incisum. We further analyzed the gene expression patterns involved in seed germination and dormancy using quantitative reverse-transcription PCR. RESULTS: RNA-sequence analysis revealed more dramatic changes in the transcriptomes of FL than those in CS, particularly for genes involved in the biosynthesis and regulation of gibberellins (GAs) and ABA. The down-regulation of ABA biosynthesis genes and the dramatic up-regulation of NiCYP707As, an ABA catabolic gene, contributed to the reduced ABA levels in FL. The increased GA3 levels in CS-treated seeds were due to the up-regulation of NiGA3OX. Both NiABI5 (a positive ABA regulator) and NiGAI (a negative regulator of GA) were down-regulated in FL and CS. The upregulation of strigolactones (SLs; the metabolites with the same precursor as ABA) biosynthesis and regulatory genes in both FL- and CS-treated seeds indicates that SLs contribute positively to seed dormancy release in N. incisum. CONCLUSIONS: Our results indicated that FL- and CS-seed dormancy release possibly depends on two totally different mechanisms: alleviation of the effects of ABA and potentiation of the effects of GA, respectively. However, NiABI5 and NiGAI probably function as common factors integrating the effects of ABA and GA on seed dormancy release.

Protein and RNA expression of acid-sensing ion channels 2 and 3 in myocardial ischemia rats induced by isoproterenol.

OBJECTIVE: To investigate the impact of electro-acupuncture at the Neiguan (PC 6) acupoint on protein and RNA expression of acid-sensing ion channel 2 (ASIC2) and ASIC3 in myocardial ischemia rats. METHODS: Fifty male Sprague-Dawley rats were used, weighing (230 ± 50) g. The rats were randomized into a normal group A, model group B, Neiguan (PC 6) group C, Lieque (LU 7) group D, and A-shi points group E. There were 10 rats in each group. Rats were continuously administered 85 mg/ kg intravenous isoproterenol daily to establish the model. Successfully modeled rats in groups C, D, and E were given electro-acupuncture treatment. Each group of rats was sacrificed with chloral hydrate (1 mL/100 g) intraperitoneal injection. The left ventricular myocardium was extracted and placed at -70 °C until use. Western blot analysis and real-time PCR were performed to assay protein and RNA expressions of ASIC2 and ASIC3, respectively. Fold changes in RNA expression were quantified with the 2^ (-ΔΔCt) method. Blood samples were drawn from the aorta abdominalis and tested for creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) levels using enzyme-linked immunosorbent assay. RESULTS: Myocardial ischemia rats given electro-acupuncture at the Neiguan (PC 6) acupoint had significantly lower protein and RNA expression of ASIC2 and ASIC3, and CK-MB and LDH levels, compared with model rats (P < 0.01). CONCLUSION: Electro-acupuncture at the Neiguan (PC 6) acupoint can not only decrease the protein and RNA expression of ASIC2 and ASIC3, but also inhibit the opening of ASICs and reduce the cardiomyocyte damage in myocardial ischemia rats.