Nonalcoholic fatty liver disease (NAFLD) is a genetic and environmental factor-associated metabolic disease that can lead to fibrosis, cirrhosis and hepatocellular carcinoma. In recent decades the prevalence of NAFLD has increased, but effective pharmacotherapy is limited. Treatment regimens in traditional Chinese medicine (TCM) have made significant contributions to the control of NAFLD, but underlying mechanisms are far less elucidated. Increasing evidence suggests that gut microbiota play a crucial role in the pathogenesis and development of diseases including NAFLD. The outcomes of such research open a new approach in identifying the molecular mechanisms of TCM. Here we review the evidence that gut microbiota might be a target in the treatment NAFLD using TCM.

¹H NMR-based metabonomic analysis was used to elucidate the hypoglycemic mechanism of Astragalus Radix and Dioscoreae Rhizomacomes. Thirty-seven SD rats were divided into four groups:model group (M group), control group (C group), Astragalus Radix and Dioscoreae Rhizomacomes group (HS group), metformin group (Y group). A T2DM model was induced with a high fat diet and streptozotocin (STZ). Drug was continuously administered for 8 weeks, after which blood and the kidneys were collected to determine the biochemical index and the kidney coefficients of each group. Using ¹H NMR metabolomics technology, we measured the metabolites in the urine of rats in each group to identify appropriate biomarkers. The results showed that total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (L-DLC), blood urea nitrogen (BUN), hemoglobin A1c (HbA1c) and the kidney coefficients were significantly increased with high density lipoprotein (H-DLC) significantly decreased in the diabetic group, but these changes were largely reversed with treatment with Astragalus Radix and Dioscoreae Rhizomacomes. A total of 20 biomarkers were found in rat urine in the diabetic group and Astragalus Radix and Dioscoreae could reverse the changes of 16 of these metabolites to varying degrees, similar to that of metformin (200 mg·kg^-1). The changes in metabolomics mainly involved butanoate metabolism, the tricarboxylic acid (TCA) cycle, taurine and hypotaurine metabolism, synthesis and degradation of ketone bodies, and pyruvate metabolism. Dioscoreae Rhizomacomes and Astragalus Radix may have a therapeutic role in the treatment of diabetes through the above five metabolic pathways, revealing the possible therapeutic mechanisms for Dioscoreae Rhizomacomes and Astragalus Radix.

A QuEChERS-ultra high performance liquid chromatography-tandem mass spectrometry method was developed for qualitative screening of 169 veterinary drug residues in bear bile powder, including β-agonists and inhibitors, antibiotics (penicillins, β-lactams, sulfomamides, quinolones, chloramphenicals, tetracyclines, nitroimidazoles, macrolides, polyethers, etc.), antiviral drugs, anthelminitics, steroid hormones, nonsteroidal antiinflammatory drugs (NSAIDs) and sedatives. The samples were extracted by Na₂EDTA-McIlvaine buffer solution and 5% fomic acid-acetonitrile solution, then purified by dispersive solid phase extraction. Detection of veterinary drug residues by ultra high performance liquid chromatography-triple quadrupole mass spectrometry was conducted and qualitative confirmed by ion ratios. The limits of detection of 169 veterinary drugs were 1-1 000 μg·kg^-1. The method is simple and fast, which had been used for the analysis of actual samples, and can be extended to the detection of similar matrix.

To prepare the mimetic exosomes and co-delivery proteins and nucleic acids, and achieve efficient and safe co-delivery of multi-component drugs, an optimized formulation was designed by modifying a polylactic acid-glycolic acid copolymer (PLGA) matrix with a cationic lipid excipient dioleyl trimethylammonium propane (DOTAP), and a PLGA/DOTAP nanoparticles packaged protein and nucleic acid was prepared by double emulsion method, and the outermost membrane structure prepared by reverse phase evaporation method and consists of 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1, 2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1, 2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol and membrane proteins. The structure of the mimetic exosomes is formed by ultrasonic dispersion and extrusion, and analyzed its characteristics and nature of the transfer effect. The size of mimetic exosomes was about 156.13 nm, with negative charge (-18.23 ±0.57 mV), and it could efficiently co-transfer protein and siRNA, and siRNA could effectively inhibit the expression of target gene Trim28. The mimetic exosomes simulate the structure of exosomes and achieve safe and efficient co-delivery of multi-component drugs.

In order to explore MYB transcription factors related to developmental processes and secondary metabolism in Morinda officinalis, we analyzed MoMYB expression based on transcriptome data from three tissues (root, stem and leaf). We used this analysis to provide a theoretical foundation for regulating the metabolism of M. officinalis. RNA-seq data along with the five databases including PFAM and plantTFDB and others were used to screen and classify MoMYB, including GO functional annotation and classification, subcellular localization, signal peptide prediction, conserved motif discovery, and comparative phylogenetic analysis. RT-qPCR was carried out to detect tissue-specific expression differences of MoMYB genes. According to transcriptome data, 109 MoMYB sequences were identified and divided into four classes, containing 51 sequences related to R2R3-MYB. Subcellular localization analysis indicated that a majority of sequences were located in nucleus. Blast2GO analysis showed that 109 MoMYB sequences were classified into three major functional ontologies including molecular function (112), biological processes (76) and cellular components (239). The R2-MYB conserved motif of 51 R2R3-MYB sequences possessed three significantly conserved tryptophan residues, whereas a phenylalanine replaced the first tryptophan in R3-MYB. The results of multiple sequence alignment and phylogenetic analysis revealed that the R2R3-MYB was distributed in all subgroups, apart from the S10, S19 and S21 subgroups. RT-qPCR indicated that several R2R3-MYB genes were differentially expressed among the three tissues, and this finding was consistent with transcriptome data. The 109 MoMYB sequences were annotated and divided into different classes, which lays the foundation for further study on MYB transcriptional factors in M. officinalis.

Melanoma is a malignant tumor with a high degree of malignancy. The incidence of melanoma keeps increasing annually. In this study, a melanoma targeted hyaluronic acid (HA) nanogel was synthesized via crosslinking of thiolated HA with terminally functionalized F127-TPGS mixed micelles. Its stability in vitro was evaluated by the average particle size, and the cytotoxicity of the nanogel was investigated by in vitro cell based assays. Next, cell uptake studies were performed to quantitatively and qualitatively investigate the uptake of the nanogels in B16F10 cells. A small sized nanogel with a diameter of 30 nm was synthesized, which was proven to be minimally cytotoxic against both 3T3 or B16F10 cells. Compared with 3T3 cells with low levels of CD44, B16F10 cells with high levels of CD44 showed significantly higher cell uptake efficiency (P < 0.05).

Plant polyphenols have a wide range of pharmacological activities and application prospects. Liquid polyphenol preparations have special physical phases and complex chemical compositions, with problems such as poor stability and easy precipitation during production and marketing. Taking the multi-precipitation mechanism of plant polyphenol liquid preparations as an example, we discuss the chemistry and composition of the precipitation, how it forms, whether precipitationcan be controlled, and the interaction law of three precipitation approaches. An unstable mechanism model is proposed where hydrolyzed tannin hydrolysis and catechin non-enzymatic oxidative polymerization repeatedly induces associative colloid aggregation and precipitation. This study explains the complex physicochemical changes in polyphenol solutions and the microcosmic mechanism of instability in the induced system and proposes a steady state reconstruction of liquid polyphenol preparation consistent with the common law of precipitation and control. It has scientific significance for promoting the development and manufacture of high quality liquid polyphenol preparations.

Two new trichilin-type limonoids were isolated from the fruits of Trichilia connaroides along with two known limonoids, 3α-deacetylamoorastatin (3) and mesendanins K (4). Their planar structure and relative configuration were elucidated by comprehensive analyses of HR-ESI-MS, ¹H NMR, ¹³C NMR, HSQC, HMBC and ROESY data. An antitumor activity assay showed that compounds 1, 2 and known compound 4 had weak cytotoxicity against a human cervical cancer cell line (HeLa).

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.

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.