Natural products (NPs) are an important source of new drugs for the treatment of stroke. Identifying cellular targets for bioactive molecules is a major challenge and critical issue in the development of new drugs for stroke. Small-molecule probes play a unique role in target discovery. However, drawbacks to these probes include non-specificity, unstable activity, and difficulty in synthesis. Small-molecule probes based on NPs at least partially compensate for these shortcomings. NPs feature rich chemical and structural diversity, biocompatibility, and unique biological activities. These features could be exploited to provide new ideas and tools for target discovery. Small-molecule probes based on NPs provide a precise and direct search for interacting protein targets of NPs-active small molecules. This review explores the properties of small-molecule probes based on NPs and their applications in mechanistic studies of stroke and other diseases. We hope that this review will bring new perspectives to the mechanistic study of NPs-active small molecules and accelerate the translation of these ingredients into drug candidates for the treatment of stroke.

In this study, 34 deep eutectic solvents (DESs) were successfully prepared for the extraction of proanthocyanidin from Rhodiolae Crenulatae Radix et Rhizomes. The extraction process was optimized using single factor exploration and Box-Behnken design-response surface analysis. The extraction rate was significantly improved when the molar ratio of choline chloride to 1,3-propanediol was 1:3.5 and the water content was 30% (V/V) in DESs. AB-8 macroporous resin and ethyl acetate were used for separation and refining, and the oligomer-rich proanthocyanidin components were eventually obtained. The ultraviolet (UV) and infrared (IR) spectra showed that the proanthocyanidins were mainly composed of catechin and epicatechin. To further clarify the chemical composition of proanthocyanidin, an ion scan list containing 156 proanthocyanidins precursors was obtained by constructing a proanthocyanidins structural library and mass defect filtering (MDF) algorithm, combined with the full mass spectrometry (MS)/dd-MS² scan mode that turns on the "if idle pick others" function. By using ultra-high performance liquid chromatography and high-resolution MS (UHPLC/HRMS), the analysis used both targeted and non-targeted methods to detect proanthocyanidins. Finally, 50 oligomeric proanthocyanidin (OPC) compounds were identified, including 7 monomers, 22 dimers, 20 trimers, and 1 tetramer, most of which were procyanidins of proanthocyanidins (84%), and a small amount of prodelphinidin (14%) and other types of proanthocyanidins (2%), which enabled the systematic characterization of proanthocyanidin components from Rhodiolae Crenulatae Radix et Rhizomes. Meanwhile, the comparison with the grape seeds OPCs standard (United States Pharmacopeia) revealed that the proanthocyanidins in Rhodiolae Crenulatae Radix et Rhizomes were more abundant, suggesting that the proanthocyanidins in Rhodiolae Crenulatae Radix et Rhizomes has promising applications.

For wild natural medicine, unanticipated biodiversity as species or varieties with similar morphological characteristics and sympatric distribution may co-exist in a single batch of medical materials, which affects the efficacy and safety of clinical medication. DNA barcoding as an effective species identification tool is limited by its low sample throughput nature. In this study, combining DNA mini-barcode, DNA metabarcoding and species delimitation method, a novel biological sources consistency evaluation strategy was proposed, and high level of interspecific and intraspecific variations were observed and validated among 5376 Amynthas samples from 19 sampling points regarded as "Guang Dilong" and 25 batches of proprietary Chinese medicines. Besides Amynthas aspergillum as the authentic source, 8 other Molecular Operational Taxonomic Units (MOTUs) were elucidated. Significantly, even the subgroups within A. aspergillum revealed here differ significantly on chemical compositions and biological activity. Fortunately, this biodiversity could be controlled when the collection was limited to designated areas, as proved by 2796 "decoction pieces" samples. This batch biological identification method should be introduced as a novel concept regarding natural medicine quality control, and to offer guidelines for in-situ conservation and breeding bases construction of wild natural medicine.

OBJECTIVE To investigate the effects of different drying methods on the index components in wine-processed Cornus officinalis so as to optimize drying method.METHODS After processed with wine， C. officinalis decoction pieces were dried with different drying methods （blast drying， far infrared drying， microwave drying， freeze drying， sun drying， shade drying and combined drying）. The contents of 5 components such as gallic acid in wine-processed C. officinalis were determined by high- performance liquid chromatography. The contents of total flavonoids in wine-processed C. officinalis were determined by chromogenic method. Analytic hierarchy process was used to evaluate the effects of different drying methods on the contents of components in C. officinalis.RESULTS The contents of gallic acid， 5-hydroxymethylfurfural， monoside， loganin， cornuside and total flavonoids in 22 batches of wine-processed C. officinalis were 1.043 8-1.563 8， 0.648 5-2.358 8， 5.031 0-10.305 7， 6.681 2- 7.534 2， 0.986 5-1.148 8 and 33.657 2-50.741 5 mg/g， respectively. The comprehensive scoring results of analytic hierarchy process showed that the comprehensive score of each component in C. officinalis dried by microwave at 75 ℃ was higher ， followed by blast drying at 60 ℃ and far infrared drying at 60 ℃ .CONCLUSIONS The wine-processed C. officinalis could be dried by microwave drying at 75 ℃， blast drying at 60 ℃ or far infrared drying at 60 ℃.

Objective To evaluate the quality of Psoralea corylifolia collected from 12 provinces of China.Methods An HPLCDAD-MS/MS method was used to identify,determine,and estimate 14 representative bioactive compounds in P.corylifolia.Then on the basis of the content data,the chemometrics method was used to differentiate 20 samples from different regions.Results The quality of P.corylifolia from 12 different provinces of China was evaluated by this method.Though the samples showed similar profiles,content of the detected markers varied significantly in different regions and batches.According to the results of the hierarchical cluster analysis and principle component analysis,it can be concluded that the samples from different origins could be clustered reasonably into two groups,as well as successfully distinguished.Conclusion A simple and reliable new method which used HPLC-DAD-MS/MS and chemometrics has been developed to characterize,classify,and control the quality of P.corylifolia.