In recent years, the prevalence of non-alcoholic fatty liver disease (NAFLD) has been dramatically increased in China and specific targeted therapy is still unavailable. The purpose of this research was to investigate whether schisandrin B (SchB) improves NAFLD and the potential mechanisms. Wildtype mice with C57BL/6J background were treated with special high fat diet (containing 40% fat, 22% fructose, 10% sucrose, and 2% cholesterol) for 16 weeks to induce NAFLD. Then SchB (120 mg·kg^-1) were used to treat NAFLD mice for 6 weeks. Body weight, food intake, glucose tolerance, insulin resistance, serum level of total cholesterol (TC), triglycerides (TG), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were assayed and histopathological analysis were performed to evaluate the improvement of NAFLD induced by SchB. Furthermore, the level of lipopolysaccharide (LPS) in serum, intestinal permeability, and the expression of genes and proteins associated with mucosal defense were evaluated, intestinal flora composition in fresh cecal contents were analyzed and differential flora were identified to explore the potential mechanism. All animal experiments were approved by the Animal Research Committee of Shanghai University of Traditional Chinese Medicine. These results showed that SchB significantly reduced the body weight of NAFLD mice without changing food intake, and effectively reduced serum level of TC, ALT, and AST. SchB also significantly altered the composition of the microflora in NAFLD mice, increased the abundance of the Akkermansia muciniphila (A. Muciniphila) and elevated the expression of genes and proteins associated with the mucosal defense in ileum and colon, restored the permeability of intestinal barrier. In summary, SchB improves NAFLD by regulating the composition of the microflora and enhancing the function of intestinal barrier to further reduce the excessive lipids accumulation and hepatic inflammation.

To explore the preventive effects and potential mechanism of mulberry leaf water extract (MLWE) on type 2 diabetes mellitus (T2DM), this study observed the influence of MLWE on the metabolism of arachidonic acid (AA). T2DM mice were induced by high fat and high sucrose (HFHS) diet and intra-peritoneal injection of streptozotocin. The mice were randomly divided into a normal control group treated with drug-free solution (NC group), a normal control group treated with MLWE (NCML group), a diabetes mellitus (DM) group treated with drug-free solution (HFHS group), and a DM group treated with MLWE (HSML group); mice were maintained on this protocol for 10 weeks. Animal experimentation was approved by the Committee on the Ethics of Animal Experiments of Xuzhou Medical University. Mice livers and plasma were collected at the end of experiment. Fasting blood glucose (FBG) and fasting blood insulin (FBI) were detected by kits. The mRNA and protein expression levels of relative metabolic enzymes related to AA in mice livers were respectively detected by quantitative real time PCR (qPCR) and Western blot. The contents of AA and its relative metabolites in mice plasma were determined by liquid phase chromatography coupled with tandem mass spectrometry. The results showed that there was no significant variation for the relative expression of metabolic enzymes, contents of FBG, FBI, AA, and its relative metabolites between NCML group and NC group. Compared with NC group, the relative expression of fatty acid synthase (Fasn), phospholipase A2 (PLA2), cyclooxygenase-2 (COX-2), and lipoxygenase-5 (LOX-5), and the levels of FBG, FBI, AA, and its relative metabolites in DM group were obviously increased, while that of carnitine palmitoyltransferase 1A (CPT1A) and cytochrome P450 family 4A (CYP4A) were significantly decreased. After intervention with MLWE, those changes could be improved, indicating that MLWE could prevent T2DM by acting on AA metabolism.

Blocking the binding of programmed death 1 (PD-1) on the T cells and programmed death ligand 1 (PD-L1) on the tumor cells has become a hotspot in the field of tumor immunotherapy. Small-molecule checkpoint inhibitor targeting PD-1/PD-L1 axis is the new direction of tumor immunotherapy. In the present study, we investigated the anti-tumor role of hyperoside by regulating the PD-L1 level in non-small cell lung cancer (NSCLC). Changes of total PD-L1 and membrane PD-L1 levels were determined by Western blot, flow cytometry, and PD-1/PD-L1 interaction assays. The expression of mRNA level of PD-L1 was detected by real-time PCR. The cytotoxicity of activated human T cells toward co-cultured tumor cells was measured by cell impedance assay and crystal violet experiment. The antitumor effect of hyperoside in vivo was examined by C57BL/6 mice bearing Lewis xenograft tumor. Western blot and flow cytometry assay showed that hyperoside significantly downregulated the abundance of PD-L1 in H1975 and HCC827 cells in dose- and time-dependent manner. PD-1/PD-L1 binding assay revealed that hyperoside reduced the binding of tumor cells to recombinant PD-1 protein. In addition, hyperoside decreased the abundance of c-Myc, a key transcriptional regulator of PD-L1, in H1975 and HCC827 cells. Cell impedance and crystal violet staining indicated that hyperoside enhanced the killing activity of co-cultured T cells toward tumor cells. Animal experiments (all animal experiments were conducted in accordance with the Animal Ethics Committee of the Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences) revealed that hyperoside treatment displayed significant suppression in the growth of Lewis tumor xenografts in C57BL/6 mice with an inhibition rate of 48.3% at 25 mg·kg^-1. Our results demonstrate that hyperoside exerts its anti-NSCLC activity by reducing the PD-L1 level. Our study provides an important material basis and scientific basis for developing hyperoside into a new small molecule drug for tumor immunotherapy.

Twenty-five quinolizidine alkaloids (including matrine-type 1-14, sparteine-type 15-17, cytisine-type 18-23, other types 24 and 25) were isolated from the roots and rhizomes of Sophora tonkinensis by various chromatographic methods. Their structures were elucidated by physicochemical properties, NMR and MS spectral data. Among them, 12-(1-acetoxyethyl)-cytisine (23) is a new alkaloid derivative, and compounds 13, 16, 17, 24 were isolated from the roots and rhizomes of S. tonkinensis for the first time. Compounds 1, 6, 19 and 20 showed potent inhibitory activity against LPS-induced NO production in RAW 264.7 macrophages, with IC₅₀ values of 39.86 ±0.65, 23.66 ±0.37, 34.56 ±0.45, 47.68 ±0.58 μmol·L^-1, respectively.

Five compounds were isolated from an ethanol extract of Artemisia deversa Diels by solvent extraction, Diaion HP-20, CHP20/P120 MCI, Sephadex LH-20, silica gel and preparative high performance liquid chromatography. Their structures were elucidated by MS, NMR and X-ray as artemideversal (1), 7-hydroxy-6-methoxycoumarin (2), 6, 7-dimethoxycoumarin (3), caffeic acid (4) and 4', 5, 7-trihydroxy flavone (5). Compound 1 is a new eudesmane-type sesquiterpene. In vitro cytotoxic activities of the five compounds were explored by MTT testing with HepG2, A549, HeLa and MRC-5 cell lines. Results show that compounds 1 and 5 significantly inhibited cellular proliferation. The IC₅₀ of compound 1 in A549 and HepG2 cells was 8.36 and 16.51 μmol·L^-1; the IC₅₀ of compound 5 in A549 and HepG2 cells was 17.06 and 7.95 μmol·L^-1.

An ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established for the simultaneous determination of seven components (chlorogenic acid, rutin, narirutin, hesperidin, sinensetin, nobiletin, tangeretin) in Citri Exocarpium Rubrum. The separation was performed over 12.0 min on a Shim-pack GIST C18 column (100 mm×2.1 mm, 2.0 μm) with a mobile phase consisting of acetonitrile (A) and 0.1% formic acid water solution (B) with gradient elution at a flow rate of 0.4 mL·min^-1; the column temperature was 30℃ and the injection volume was 1 μL. Detection was performed on a QTRAP 4500 mass spectrometer equipped with electrospray ionization. Positive and negative electrospray ionization was performed in multiple reaction monitoring mode. All of the analytes showed good linearity (r ≥ 0.997 3) in the tested ranges, with good precision, repeatability and stability for all analytes. The average recoveries were in the range of 99.32%-107.75% with relative standard deviations RSD ≤ 3.94%. The established method is accurate, stable and reproducible, and can provide a research method for the quality control of Citri Exocarpium Rubrum.

Type 2 diabetes is a common form of diabetes and can have serious consequences for diabetics when their wounds do not heal properly due to impaired function. Glycosaminoglycans (GAGs) are widely found in skin tissue. Due to the isomerization of C6 in uronic acid and different sulfuric acid substitutions, the structure of GAGs is relatively complex. The fine structure of GAGs in the skin of diabetic patients has not been studied. In this study, the structure of GAGs in the skin of streptozotocin (STZ)-induced diabetic mice was characterized by liquid chromatographytandem mass spectrometry (LC-MS/MS). All animal experiments were carried out with approval of the Animal Ethics Committee of Ocean University of China. The results indicate that the content of hyaluronic acid (HA) in STZ-induced diabetic mouse skin was significantly lower than that of non-diabetic mice. Although there was no significant difference in the content of chondroitin sulfate (CS) and heparin sulfate (HS) between diabetic mouse and normal mouse skin, the disaccharide compositions were different. The expression of CS-4S6S, HS-0S, HS-NS, and HS-6S in STZ-induced diabetic mouse skin was higher than in non-diabetic mice. While the content of HS-NS6S was lower. In addition, the degree of sulfation of HS in STZ-induced mouse skin was lower than that of normal mouse skin. These results provide a basis for the pathogenesis of diabetes and the development of wound healing dressings for diabetic patients.

The metabonomics method was used to explore the processing and synergistic mechanism of epimedium fried with suet oil in warming the kidney and enhancing yang. The kidney-yang deficiency rat model was established by injection of hydrocortisone. Then the UPLC-Q-TOF-MS (ultra-performance liquid chromatography with quadrupole time-of-flight tandem mass spectrometry) metabolomics method was combined with multivariate statistical analysis methods and univariate statistical analysis to screen and identify kidney-yang deficiency potential biomarkers in plasma and urine samples. Finally the metabolic regulation mechanism of suet oil group, the epimedium raw product group, the epimedium heating product group, and the epimedium fried with suet oil group improved kidney-yang deficiency was analyzed. The results showed that the plasma and urine metabolism of rats with kidney-yang deficiency induced by hydrocortisone showed obvious trajectory changes. 15 biomarkers related to kidney-yang deficiency were identified in plasma and urine, involving 5 metabolic pathways, namely glycerophospholipid metabolism, sphingolipid metabolism, sulfur metabolism, glyoxylate acid and dicarboxylate metabolism, and cysteine and methionine metabolism. The metabolic pathway of epimedium fried with suet oil warming kidney and promoting yang involved glycerophospholipid metabolism, cysteine and methionine metabolism, and the two processing factors of epimedium fried with suet oil "heating" and "suet oil" enhanced its function of warming the kidney and promoting yang by regulating glycerophospholipid metabolism, cysteine and methionine metabolism, respectively. In this way, the processing and synergistic mechanism of epimedium fried with suet oil was clarified. The animal experiments involved in this article comply with ethical standards and have been approved by the Animal Ethics Committee of Jiangsu Provincial Academy of Chinese Medicine (approval number: AEWC-20200702-119).

Flavanone 3-hydroxylase (F3H), flavonol synthase (FLS) and anthocyanin synthase (ANS) are members of the 2-oxoglutarate-dependent oxygenase (2-ODD) superfamily, which play important functions in plant flavonoid biosynthesis. In this study, 16 2-ODD genes involved in flavonoid biosynthesis pathway were screened and identified from safflower genome database using bioinformatics method. The length of the proteins encoded by these genes ranged from 283 (CtFLS2) to 442 aa (CtANS6), and the molecular weight ranged from 32 062.05 (CtFLS2) to 48 245.49 Da (CtANS6). The proteins encoded by these genes were hydrophilic. Phylogeny divided it into three subfamilies, two conserved residues, H-x-D-xn-H and R-x-S, were found in this gene family using conserved motifs analysis, the analysis of cis-acting elements in the upstream 2 000 bp region showed that these genes might be regulated by environmental factors such as light, temperature and plant hormones such as salicylic acid, methyl jasmonate and so on. The members of the safflower 2-ODD gene family involved in flavonoid biosynthesis pathway were analyzed by qRT-PCR, the result showed that the expression patterns of these members were different in diverse flowering period and leaf. This study can lay a foundation for further exploring the function of 2-ODD genes involved in flavonoid biosynthesis pathway in safflower.

Twenty-five candidate MLO genes of Lonicera japonica were obtained by whole genome sequencing. Bioinformatics analysis showed that the number of amino acids in the protein family ranged from 137 to 846, the theoretical isoelectric point ranged from 5.02 to 9.50, and it was rich in basic amino acids. One protein did not contain transmembrane domain, and the other proteins ranged from 3 to 10. The results of subcellular localization showed that 21 proteins were located on the cell membrane and 1 protein was located on the chloroplast. Phylogenetic trees were constructed from 133 MLO proteins of Lonicera japonica, Triticum aestivum, Arabidopsis thaliana, Solanum lycopersicum, Nicotiana tabacum, Nicotiana sylvestris and Nicotiana tomentosiformis. The results indicated that MLO family proteins of Lonicera japonica could be divided into five subgroups. Tissue specific analysis showed that the expression of MLO genes in Lonicera japonica had obvious tissue specificity. Among them, 8 genes were highly expressed in leaves, 2 genes in stems and 2 genes in flowers. Four genes were significantly upregulated after inoculation with powdery mildew, among which MLO14 increased the most, which was more than 2 000 times higher than that of the control. In this study, the composition and expression of the MLO gene related to the occurrence of powdery mildew of Lonicera japonica were preliminarily analyzed, which laid a foundation for the further use of MLO as a target gene to develop new germplasm resistant to powdery mildew of Lonicera japonica.