This paper describes a bioassay method for the determination of ansamitocin titers. A fungal strain sensitive to ansamitocin was classified to the genus Trichoderma based on phylogenetic analysis of its ITS sequence, and designated as Trichoderma CPCC 400749. PDA plates of Trichoderma CPCC 400749 were prepared to assay ansamitocin titers of Actinosynnema pretiosum ATCC 31565. The titers were consistent with those determined by HPLC. The bioassay method may have the potential use in high-throughput screening for Actinosynnema pretiosum mutants with improved ansamitocin titers.

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 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.

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.

In recent years, chimeric antigen receptor T cells (CAR-T) have been viewed as a target for successful treatment of hematologic malignancies. However, targeting conventional CAR-T cell has a series of side effects, such as cytokine storm, on-target off-tumor effect and neurotoxicity during treatment, and these side effects threatened patients' life. The extracellular domain of conventional CAR-T is a fixed single-chain variable fragment (scFv) that only targets one specific antigen, and once the tumor antigen is mutated or disappears, the CAR-T cell will fail. In recent years, a number of different switchable CAR-T cells have emerged. The design of switchable CAR-T cells is divided into two aspects:CAR-T cell and molecular switch respectively, and the activation of CAR-T is completely dependent on the switch. It is not only universal, but also decreases the side effect of conventional CAR-T through controlling the molecular switch. We summarized the existing sCAR-T to provide an idea for CAR-T design and optimization, and lay a foundation for entering sCAR-T into clinical practice.

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.

In this study, the lipid membrane-wrapped nanoparticles loaded with metformin polymer (PolyMet) and doxorubicin (DOX) was prepared and then evaluated therapeutic effect on breast cancer. An anionic chain PGA-DOX based on γ-polyglutamic acid (PGA) with DOX was synthesized via amidation reaction and characterized by ¹H NMR. The PGA-DOX and PolyMet were loaded via electrostatic attraction to prepare the co-delivery nanoparticles system (PolyMet-DOX-NPs). Then, PolyMet-DOX-NPs were coated with cationic liposome membrane to form the core-membrane structural system (PolyMet-DOX-lipid-nanoparticles, PolyMet-DOX-LNPs). The structure and morphology of PolyMet-DOX-LNPs were observed by transmission electron microscope. The particle size, zeta potential, encapsulation efficiency (EE), drug loading (DL), release behavior in vitro of PolyMet-DOX-LNPs were investigated. The MTT assay was used to examine the cytotoxicity of PolyMet combined with DOX on 4T-1 cells. The 4T1^Fluc tumor-bearing mice model was used to evaluate the therapeutic efficacy of PolyMet-DOX-LNPs in vivo. All animal experiments were performed in line with ethical standards and approved by the Animal Experiments Ethical Committee of Zhejiang Chinese Medical University. ¹H NMR spectrum showed that PGA-DOX was successfully synthesized with DOX grafting rate of (72.03 ±1.29)%. The EE and DL of PolyMet-DOX-LNPs was (72.76 ±1.92)% and (1.16 ±0.12)%, respectively. PolyMet-DOX-LNPs exhibited a suitable size of (159.3 ±7.4) nm and positive charge of (+36.3 ±1.9) mV with good spheroidal morphology and dispersibility. The release profiles in vitro showed that PolyMet-DOX-LNPs exhibited a slowly and maintained release behavior at physiological pH value (pH 7.4) within 48 h. Further studies showed that PolyMet combined with DOX could synergistically enhance the cytotoxicity on 4T-1 cells. Bioluminescence imaging (BLI) result showed that the luminescence signal intensity of 4T-1^Fluc cells was reduced after treatment with PolyMet-DOX-LNPs and the tumor volume growth was also inhibited. Additionally, the H&E staining and changes of body weight showed that PolyMet could reduce the toxicity of DOX. To sum up, PolyMet has a good synergistic effect with DOX in the treatment of breast cancer, which provide the foundation for this novel metformin polymer on the anti-tumor application.

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.

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.

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.