Based on the ITS2 and psbA-trnH sequences, molecular biological identification and genetic relationship of Fritillaria cirrhosa with its relative species were carried out. In this paper, the PCR-RFLP method specified by the Chinese Pharmacopoeia was performed on all samples at first. Secondly, the ITS2 and psbA-trnH sequences of all samples were amplified. Then, the amplified products were used to analyze the genetic distance, construct the phylogenetic tree, assess the identification efficiency, and evaluate the genetic relationship as well. The result showed that all the samples were divided into two groups by PCR-RFLP method. The samples in the first group, including Fritillaria ussuriensis, Fritillaria thunbergii and Fritillaria pallidiflora, could not be digested by SmaI, while the other samples in the second group, including Fritillaria mellea, Fritillaria sinica, Fritillaria cirrhosa var. ecirrhosa Franch, Fritillaria unibracteata var. longinectarea and Fritillaria cirrhosa, could be digested by SmaI. Then, ITS2 and psbA-trnH sequences of all samples were obtained. The length of various ITS2 sequences were distributed from 235 to 239 bp, and the average intra-and inter-specific genetic distance were 0.001 and 0.022, respectively. NJ tree showed that all samples were separated into "Northern Fritillaria" group (Fritillaria ussuriensis and Fritillaria pallidiflora) and "Southern Fritillaria" group (Fritillaria thunbergii, Fritillaria mellea, Fritillaria sinica, Fritillaria cirrhosa var. ecirrhosa Franch, Fritillaria unibracteata var. longinectarea and Fritillaria cirrhosa). The latter group could be further divided into Fritillaria thunbergii and Fritillaria cirrhosa subgroup, and the species in Fritillaria cirrhosa subgroup had close phylogenetic relationships. The length of psbA-trnH sequences was distributed from 337 to 373 bp, and the intra-and inter-specific genetic distance were 0.263 and 0.329, respectively. The samples in this paper could not be clustered effectively by NJ tree. This indicated that the ITS2 sequences were not only able to identify Fritillaria cirrhosa with its partial relative species quickly and accurately, but also clarify the relationship between different Fritillaria species. Therefore, it provided an important theoretical foundation for the development of molecular markers, effective protection, and rational development and utilization of Fritillaria resources.

To obtain the microbial composition of traditional Chinese medicine of Faeces Trogopterori, ten samples were collected from the imitate wildness farmland in Shangluo City, Shaanxi Province. In this study, 16S rRNA gene was used as molecular marker to explore the microbiome and the sequences were analyzed by Usearch analysis platform. The COG and KEGG database is used to predict and analyze the function of the flora. A great number of 285 218 high quality clean reads with a length of 400-450 bp were obtained from 10 samples. Bacterial species detected in these samples covered 8 phyla, 25 families, 75 genera and 120 species. The dominant phylum microbial communities in these samples were Firmicutes (87.68% ±2.68%) and the Bacteroidetes (7.62% ±3.74%), all samples showed a high microbial diversity, the predicted functional metagenome was heavily involved in energy metabolism. This study provided that the beneficial bacteria in Faeces Trogopterori may be one of its active ingredients, and no pathogens are detected in the sample.

We study here in vitro cytotoxicity, in vivo tumor inhibition and the mechanism on photodynamic therapy (PDT) of photosensitizer BF01 using human hepatocellular carcinoma cell line BEL-7402. CCK-8 method was used to detect the inhibition rate and IC₅₀ in BEL-7402 cells on the same laser intensity with varying concentrations (0, 0.8, 1.6, 3.2, 6.4 μmol·L^-1) of photosensitizer BF01. Cell death mode of BEL-7402 was detected by flow cytometry, with apoptotic characteristics observed by DAPI staining, and the subcellular localization of reactive oxygen was observed using photodynamic detection and confocal microscopy. The cell model of human liver cancer in nude mice was established, tumor growth curve was drawn, and the therapeutic effect of BF01 was determined. The animal experimentation was approved by East China University of Science and Technology Ethics Committee. The results indicated that BF01 PDT treatment can clearly inhibit BEL-7402 tumor cell proliferation, with the killing rate of 86% at the concentration of 6.4 μmol·L^-1 of BF01, and half lethal concentration IC₅₀ value of 2.46 μmol·L^-1. DAPI stained nuclei shows the characteristics of advanced stage apoptosis, whereas reactive oxygen species level in the mitochondria increased with increasing drug concentration. In vivo experiments showed that photosensitizer BF01 mediated photodynamic therapy of liver cancer cells and inhibited tumor growth in mice. Therefore, the new BF01 photosensitizer has a potential for development into future clinic application.

We have developed a new method using HPLC-CAD (charged aerosol detector) for the quantitative analysis of cyclovirobuxine D and related substances in the API of Huangyangning tablets. The related substances were further studied by HPLC-Q-Exactive coupled with hybrid quadrupole-orbitrap mass spectrometry. A HILIC column of XBridge Amide (4.6 mm×250 mm, 5 μm) was used, and the mobile phase was composed of acetonitrile and 100 mmol·L^-1 ammonium formate (85:15), which was adjusted to pH 2.8 with formic acid. Isocratic mode elution was adopted at a flow rate of 1.1 mL·min^-1. The column temperature was set at 30℃. For CAD, the temperature of atomization and gas pressure were respectively set at 35℃ and 62.2 psi. This method detected and quantified five related substances to cyclovirobuxine D. The results showed that the LOD and LOQ of cyclovirobuxine D was 12.588 ng and 28.323 ng, respectively with an average recovery of 95.74% (RSD=1.79%, n=6). The content of cyclovirobuxine D in 12 batches of API samples provided by three manufacturers was from 79.94% to 88.49%, with an average value of 82.20%. The total content of the five related substances was from 15.99% to 22.15% with an average value of 20.10%, using an external standard method with cyclovirobuxine D as the reference and according to the CAD uniform response to non-volatile substances. The newly developed HPLC-CAD method has advantages in terms of the comprehensiveness of signals from Buxus alkaloids without UV absorption and with high sensitivity to its trace-related substances; the method yields good separation between the components and is compatible with mass spectrometry. It is applicable for the accurate quantitative analysis of main components and related substances in the API of Huangyangning tablets.

We established a quality evaluation method for Shuanghuanglian preparations based on an effect-constituent index (ECI), which is guided by the clinical efficacy of Shuanghuanglian and a dose-efficacy correlation. An HPLC method was used to establish the quantitative fingerprint of Shuanghuanglian from different manufacturers and to determine the content of 10 fingerprint components, including baicalin, chlorogenic acid, forsythin, galuteolin, wogonin, forsythoside A, luteolin, caffeic acid, baicalein, and scutellarin. Using Staphylococcus aureus as biological model, the potency of Shuanghuanglian preparations was determined by antibiotic microbial assay. Using the method of PLC-DA, the efficacious antibacterial components were screened by "dose-efficacy" correlation analysis. According to the antibacterial potency and content of the antibacterial ingredients, combined with the method of the custom weight coefficient, ECI was calculated and verified. The results show that the antibacterial ECI can facilitate evaluation of the efficacy of Shuanghuanglian based on the composition of its contents, providing a new method for the quality control of traditional Chinese medicine.

Injectable traditional Chinese medicine often contains multiple components including undefined toxic substances, can have high variability between batches, with undefined mechanisms of action. It is urgent to improve the quality and consistency and reduce the toxicity risk of traditional Chinese medicine. The Microtox technology is a simple, rapid method for the detection of toxic substances in the environment that uses non-pathogenic luminescent bacteria as an indicator, and the change in luminosity as an index. Using this bioassay we have systematically applied Microtox technology for the detection of microtoxicity in injectable traditional Chinese medicine. As a new method of bioactivity characterization, Microtox technology is expected to be used in the detection of quality fluctuations and toxicity risks at an early stage in the preparation of injectable traditional Chinese medicines and to improve the quality of injectable traditional Chinese medicine.

One undescribed and two known furocoumarins were isolated from the stems of the Clausena lansium through a series of isolation and purification approaches including HPD-100 macroporous resin column, silica gel, reverse phase C₁₈ and so on. Their structures were determined to be 8-[(2S, 3S, 6E)-2, 3-epoxy-3, 7-dimethyl-oct-6-enyloxy] psoralen (1), 8-(7', 8'-epoxygeranyloxy) psoralen (2) and 8-[(2E)-6-oxo-3, 7-dimethyloct-2-enyloxy] psoralen (3) by spectroscopic methods. Compound 1 is a new furocoumarin. Compound 2 showed cytotoxicity to H460 (IC₅₀=43.94 μmol·L^-1) and compound 3 showed cytotoxicity to HeLa (33.76 μmol·L^-1) through the cytotoxic tests against five human cancer cell lines (H460, H7402, HCT-8, HeLa and MCF-7) for all compounds.

Phytochemical study of the aerial parts of Hypericum perforatum L. resulted in the isolation of an undescribed compound, which was identified as Rel-(2S, 3R)-2-(3, 4-dihydroxyphenyl)-3, 5, 7-trihydroxy-2-methoxy-3-(2-oxopropyl)chroman-4-one (1) by spectroscopic methods including UV, IR, HR-ESI-MS, 1D and 2D NMR spectra. Compound 1 is a new 2, 3-dioxo-flavone with an acetonyl moiety, rarely found in nature. In addition, a plausible biogenetic pathway of 1 was proposed in this article.

To analyze the chemical profile of Radix Bupleuri-Radix Paeoniae Alba in the serum, after oral absorption of the herbal pair, we used UPLC-QE-Orbitrap-MS technique and Compound Discover 2.0 software to compare the components in the extracts, and drug-containing serum. The map, based on the precise molecular weight and secondary fragment information given by the mass spectrometry, combined with the reference substance and reference information for, finding the chemical components in the blood after oral administration of the herbal pair in the rats. Animal experimentation was approved according to the Committee on the Ethics of Animal Experiments of Shanxi University. The results showed that a total of 55 components were detected in the serum, of which 16 were prototype components and 39 were metabolites of prototype components. UPLC-QE-Orbitrap-MS allows quick, accurate and comprehensive analysis of the blood components of the herb pair after oral administration. This preliminary study of the chemical components in the blood after herb pair ingestion provides a basis for the subsequent research on the pharmacodynamics and drug-drug interactions.

Consistency in quality of traditional Chinese medicine granules is an important factor to ensure reproducible clinical efficacy. In this study rhubarb dispensing granules were utilized to construct an efficacious near-infrared spectroscopy (eNIRS) assay by combining NIRS and biopotency. A NIR method for assaying rhubarb dispensing particles was established, and information on different batches was collected. The diarrhea-inducing biopotency of rhubarb dispensing granules was determined based on a constipation model induced by diphenoxylate in mice. The animal protocol was approved by the Animal Ethic Committee of 302 Hospital of Chinese PLA People's Liberation Army (ID:IACUC-2019-0010). Ten anthraquinones were determined in rhubarb dispensing granules by UPLC. The correlation between NIR and biopotency was analyzed and five characteristic bands that correlated highly with bioactivity were identified, including 4 011-4 390, 4 859-5 461, 7 012-7 493, 10 992-11 312 and 11 871-12 489 cm^-1. There were some differences in the main bands of different chemical constituents. In summary, five active bands based on NIRS were identified and found to be able to achieve rapid on-line detection of rhubarb dispensing granule quality. This research model may also provide reference for quality control of other Chinese medicine dispensing granules.