Utilizing network pharmacology, molecular docking, and animal models to explore the therapeutic potential of the WenYang FuYuan recipe for cerebral ischemia-reperfusion injury through AGE-RAGE and NF-κB/p38MAPK signaling pathway modulation.

BACKGROUND: Stroke is a debilitating condition with high morbidity, disability, and mortality that significantly affects the quality of life of patients. In China, the WenYang FuYuan recipe is widely used to treat ischemic stroke. However, the underlying mechanism remains unknown, so exploring the potential mechanism of action of this formula is of great practical significance for stroke treatment. OBJECTIVE: This study employed network pharmacology, molecular docking, and in vivo experiments to clarify the active ingredients, potential targets, and molecular mechanisms of the WenYang FuYuan recipe in cerebral ischemia-reperfusion injury, with a view to providing a solid scientific foundation for the subsequent study of this recipe. MATERIALS AND METHODS: Active ingredients of the WenYang FuYuan recipe were screened using the traditional Chinese medicine systems pharmacology database and analysis platform. Network pharmacology approaches were used to explore the potential targets and mechanisms of action of the WenYang FuYuan recipe for the treatment of cerebral ischemia-reperfusion injury. The Middle Cerebral Artery Occlusion/Reperfusion 2 h Sprague Dawley rat model was prepared, and TTC staining and modified neurological severity score were applied to examine the neurological deficits in rats. HE staining and Nissl staining were applied to examine the pathological changes in rats. Immunofluorescence labeling and Elisa assay were applied to examine the expression levels of certain proteins and associated factors, while qRT-PCR and Western blotting were applied to examine the expression levels of linked proteins and mRNAs in disease-related signaling pathways. RESULTS: We identified 62 key active ingredients in the WenYang FuYuan recipe, with 222 highly significant I/R targets, forming 138 pairs of medication components and component-targets, with the top five being Quercetin, Kaempferol, Luteolin, ß-sitosterol, and Stigmasterol. The key targets included TP53, RELA, TNF, STAT1, and MAPK14 (p38MAPK). Targets related to cerebral ischemia-reperfusion injury were enriched in chemical responses, enzyme binding, endomembrane system, while enriched pathways included lipid and atherosclerosis, fluid shear stress and atherosclerosis, AGE-RAGE signaling in diabetic complications. In addition, the main five active ingredients and targets in the WenYang FuYuan recipe showed high binding affinity (e.g. Stigmasterol and MAPK14, total energy <-10.5 Kcal/mol). In animal experiments, the WenYang FuYuan recipe reduced brain tissue damage, increased the number of surviving neurons, and down-regulated S100ß and RAGE protein expression. Moreover, the relative expression levels of key targets such as TP53, RELA and p38MAPK mRNA were significantly down-regulated in the WenYang FuYuan recipe group, and serum IL-6 and TNF-a factor levels were reduced. After WenYang FuYuan recipe treatment, the AGE-RAGE signaling pathway and downstream NF-kB/p38MAPK signaling pathway-related proteins were significantly modulated. CONCLUSION: This study utilized network pharmacology, molecular docking, and animal experiments to identify the potential mechanism of the WenYang FuYuan recipe, which may be associated with the regulation of the AGE-RAGE signaling pathway and the inhibition of target proteins and mRNAs in the downstream NF-kB/p38MAPK pathway.

Conversion of nitrogen and carbon in enriched paddy soil by denitrification coupled with anammox in a bioelectrochemical system.

The aim of this study is to investigate conversion of nitrogen and COD in enriched paddy soil by nitrification coupled with anammox process in a dual chamber bioelectrochemical system. The paddy soil was enriched for denitrification coupled with anammox by microbial consortia and was acclimatized in the cathodic chamber of microbial fuel cells (MFCs). The bioelectrochemical systems were treated with different ammonium concentrations in the cathodic chamber: the MFC with low concentration ammonium (LA-MFC, 50 mg/L ammonium), the MFC with medium concentration ammonium (MA-MFC, 500 mg/L ammonium), and MFC with high concentration ammonium (HA-MFC, 1000 mg/L ammonium), and the initial COD in the anodic chamber was 1200 mg/L. The CK treatments were conducted with 1000 mg/L ammonium under the same conditions, except without inoculum in the cathode chamber. The consumption rate of ammonium in the cathodic chambers of CK, LA-MFC, MA-MFC, and HA-MFC were 9%, 64%, 84%, and 84%, respectively. The degradation rate for COD achieved in the anode chambers of CK, LA-MFC, MA-MFC, and HA-MFC were 70%, 86%, 93%, and 93%, respectively. The analysis of the microbial community of three treated MFCs in the cathode chamber indicated that the nitrification-denitrification process occurs in the cathode chamber. The dominant species for nitrification was Nitrospira, and the dominant species for denitrification were Denitratisoma, Dechloromonas, and Candidatus_Competibacter. Moreover, anammox process also observed in the cathode chamber. The functional genes nirS/K, hzsB, and 16S rDNA were assessed by qPCR analysis, and the results confirmed the presence of denitrification-coupled anammox in the cathodic chamber.

Research on the circular RNA bioinformatics in patients with acute myocardial infarction.

OBJECTIVE: Through the detection of circular RNA (circRNA) using expression profiling chips, we searched for circRNAs related to acute myocardial infarction (AMI) and explored their relationship and possible mechanisms with AMI. METHOD: The study subjects included 3 AMI patients and 3 controls, and circRNA expression profiling analysis was performed using a microarray gene chip to identify circRNAs with large differences in expression between groups and to construct a circRNA-miRNA network. RESULTS: Compared with the control group, there were 650 differentially expressed circRNAs found in AMI patients (P < .05, fold change > 2), including 535 up-regulated circRNAs, such as hsa_circ_0050908, hsa_circRNA4010-22, hsa_circ_0081241, hsa_circ_0010551, hsa_circRNA4010-20, hsa_circRNA14702, hsa_circ_0115392, has_circRNA1825-44, has_circRNA8493-7, and hsa_circ_0025097. Furthermore, there were 115 down-regulated circRNAs, such as hsa_circ_0066439, hsa_circ_0054211, hsa_circ_0095920, hsa_circ_0122984, hsa_circ_0113067, hsa_circ_0039155, hsa_circRNA4014-45, hsa_circ_0122979, hsa_circ_0059665, and hsa_circ_0009319. The circRNAs hsa_circ_0066439, hsa_circ_0081241, and hsa_circ_0122984 can regulate multiple signal pathways to participate in the AMI process through hsa-miR-1254, hsa-miR-328-5p, and other miRNAs. In addition, the expression of circRNA-miRNA in peripheral blood is related to the network. Differentially expressed circRNAs are involved in chromatin organization, chromatin-modifying enzymes, signal transduction, lysine degradation, the mitogen-activated protein kinase (MAPK) signaling pathway, focal adhesion, and a variety of other pathways, such as myocardial infarction, coronary heart disease, hypertension, and other diseases. The gene ontology analysis results show that molecular function mainly involves binding and molecular structural activity, whereas the biological process mainly involves a single biological process, a cellular component for organization, and a cellular process, and the cellular component mainly involves a protein complex, an extracellular matrix, and a membrane. CONCLUSION: circRNA and microRNA interact to participate in the development of AMI. circRNA may be involved in the pathogenesis of AMI.

Application of Non-Invasive Prenatal Tests in Serological Preclinical Screening for Women with Critical-Risk and Low-Risk Pregnancies but Abnormal Multiple of the Median Values.

BACKGROUND: To explore the clinical value of secondary screening using noninvasive prenatal testing (NIPT) for women with critical-risk and low-risk pregnancies who had multiple of the median (MoM) abnormalities in serological screening. METHODS: NIPT was used to analyze fetal free DNA in the peripheral blood of 2,325 women with critical-risk pregnancies and 239 women with low-risk pregnancies with MoM abnormalities in serological screening. Based on NIPT results, women with high-risk pregnancies were recommended for amniocentesis for fetal karyotype analysis. RESULTS: Among 2,325 women with critical-risk pregnancies as determined by serological screening, NIPT indicated 15 high-risk pregnancies (11 cases of trisomy 21 and 4 cases of trisomy 18). Of the 15 patients, 1 case refused prenatal diagnosis. The other 14 cases underwent invasive amniocentesis for fetal karyotype analysis, and 13 cases of fetal chromosomal abnormalities were diagnosed, including ten cases of trisomy 21 and three cases of trisomy 18. NIPT of 239 patients with low-risk pregnancies but abnormal MoM showed one case with a high risk of trisomy 21, which was diagnosed as a false positive by amniotic fluid karyotype analysis, and one case with a high risk of trisomy 13 (stillbirth) that was diagnosed by karyotype analysis. A case of gender chromosome abnormality was diagnosed as aneuploidy by karyotype analysis. CONCLUSIONS: The application of NIPT as a secondary screening for women with low- and critical-risk pregnancies as determined by serological screening but with MoM abnormalities will greatly reduce the number of invasive prenatal diagnosis procedures, significantly improve the rate and accuracy of fetal chromosomal abnormality detection.