Radix Polygoni Multiflori (PM) is a well−known nootropic used in traditional Chinese medicine (TCM). Considering the efficacy and application discrepancy between raw (RPM) and processed PM (PPM), the similarities and differences between them in the treatment of vascular dementia (VaD) is intriguing. In this study, a VaD rat model was constructed by 2−vessel occlusion (2−VO). During 28 days of treatment, plasma was collected on days 7, 14, 21, and 28 after the start of dosing and the metabolic profile was analyzed by HPLC−MS/MS−based metabolomics.The Morris Water Maze Test, hematoxylin–eosin and Nissl staining, and biochemical analysis were used to assess cognitive function, pathogenic alterations and oxidative stress, respectively. RPM and PPM effectivelyreducedthe 2VO−induced cognitive impairment and mitigated histological alterations in hippocampus tissue. The 2−VO model significantly elevated MDA level and decreased SOD activity and GSH level, indicating severe oxidative stress, which could also be attenuated by RPM and PPM treatment. RPM outperformed PPM in decreasing MDA levels while PPM outperformed RPM in increasing GSH levels. Differential metabolites were subjected to Metabolite Set Enrichment Analysis (MSEA) and genes corresponding to proteins having interactions with metabolites were further annotated with Gene Ontology (GO). Both RPM and PPM ameliorated VaD−relevant vitamin B6 metabolism, pentose phosphate pathways, and taurine and hypotaurine metabolism. In addition, the metabolism of cysteine and methionine was regulated only by RPM, and riboflavin metabolism was modulated only by PPM. The results suggested that raw and processed PM had comparable efficacy in the treatment of VaD but also with some mechanistic differenece.
The zebrafish (Danio rerio) is widely used as a promising high-throughput model organism in neurobehavioral research. The mobility of zebrafish can be dissected into multiple behavior endpoints to assess its neurobehavioral performance. However, such facilities on the market are expensive and clumsy to be used in laboratories. Here, we designed a low-cost, automatic zebrafish behavior assay apparatus, barely without unintentional human operational errors. The data acquisition part, composed of Raspberry Pi and HQ Camera, automatically performs video recording and data storage. Then, the data processing process is also on the Raspberry Pi. Water droplets and inner wall reflection of multi-well cell culture plates (used for placing zebrafish) will affect the accuracy of object recognition. And during the rapid movement of zebrafish, the probability of zebrafish tracking loss increased significantly. Thus, ROI region and related thresholds were set, and the Kalman filter algorithm was performed to estimate the best position of zebrafish in each frame. In addition, all functions of this device are realized by the custom-written behavior analysis algorithm, which makes the optimization of the setup more efficient. Furthermore, this setup was also used to analyze the behavioral changes of zebrafish under different concentrations of alcohol exposure to verify the reliability and accuracy. The alcohol exposure induced an inverted U-shape dose-dependent behavior change in zebrafish, which was consistent with previous studies, showcasing that the data obtained from the setup proposed in this study are accurate and reliable. Finally, the setup was comprehensively assessed by evaluating the accuracy of zebrafish detection (precision, recall, F-score), and predicting alcohol concentration by XGBoost. In conclusion, this study provides a simple, and low-cost package for the determination of multiple behavioral parameters of zebrafish with high accuracy, which could be easily adapted for various other fields.
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