(±)-Salvicatone A: A Pair of C27-Meroterpenoid Enantiomers with Skeletons from the Roots and Rhizomes of Salvia castanea Diels f. tomentosa Stib.

(±)-Salvicatone A (1), a C27-meroterpenoid featuring a unique 6/6/6/6/6-pentacyclic carbon skeleton with a 7,8,8a,9,10,10a-hexahydropyren-1 (6H)-one motif, was isolated from the roots and rhizomes of Salvia castanea Diels f. tomentosa Stib. Its structure was characterized by comprehensive spectroscopic analyses along with computer-assisted structure elucidation, including ACD/structure elucidator and quantum chemical calculations with 1H/13C NMR and electronic circular dichroism. Biogenetically, compound 1 was constructed from decarboxylation following [4 + 2] Diels-Alder cycloaddition reaction between caffeic acid and miltirone analogue. Bioassays showed that (-)-1 and (+)-1 inhibited nitric oxide production in lipopolysaccharide-induced RAW264.7 macrophage cells with an IC50 value of 6.48 ± 1.25 and 15.76 ± 5.55 µM, respectively. The structure-based virtual screening based on the pharmacophores in ePharmaLib, as well as the molecular docking and molecular dynamics simulations study, implied that (-)-1 and (+)-1 may potentially bind to retinoic acid receptor-related orphan receptor C to exert anti-inflammatory activities.

Two new sesquiterpenoids with glycosides from the leaves and stems of Schisandra chinensis (Turcz.) Baill.

Two previously undescribed glycosidic bisnorsesquiterpenoids A - B (1 - 2), together with two known compounds (3 - 4), were isolated from the leaves and stems of Schisandra chinensis. Their structures were elucidated by extensive spectroscopic data (1D, 2D NMR, and HRESIMS). The anti-inflammatory activity, ABTS+ radical scavenging activity, and DPPH radical scavenging activity of compounds 1 - 4 were tested. However, all of these compounds showed only weak anti-inflammatory or antioxidant effects.

Wunorlactones A-G, seven undescribed 7/8/5 and 7/8/3 carbon skeleton schinortriterpenoids from the stems and leaves of Schisandra chinensis, and their neuroprotective activities.

Schisandra chinensis is an important traditional Chinese medicine and its main bioactive components are lignans and schinortriterpenoids (SNTs). The aim of this study was to explore the biologically rich SNTs from the stem and leaves of S. chinensis (SCSL). Here, seven previously undescribed 7/8/5 and 7/8/3 carbon skeleton SNTs (1-7) were reported. Their structures were determined by NMR, UV, MS, ECD, and X-ray diffraction analyses, and the neuroprotective activities of these compounds on corticosterone-induced PC12 cell injury were evaluated. The results showed that 1, 5, and 7 (25 µM) had neuroprotective effects, and the cell viability was increased by 20.07%, 14.24%, and 15.14% (positive control: 30.64%), respectively. These findings increased the number of described SNTs in SCSL, and the neuroprotective activities of all compounds indicated their potential application in neurodegenerative diseases.

An integrated network pharmacology approach reveals that Darutigenol reduces inflammation and cartilage degradation in a mouse collagen-induced arthritis model by inhibiting the JAK-STAT3 pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Darutigenol (DL) is a natural active product derived from the Chinese herbal medicine Sigesbeckia glabrescens (Makino) Makino. It is administered as a traditional Chinese medicine (TCM) to dispel rheumatism, benefit the joints, and detoxify. However, its potential mechanism in the treatment of rheumatoid arthritis (RA) remains unknown. AIMS OF THE STUDY: The objectives of this research were to determine the effects and elucidate the modes of action of DL on RA-related joint inflammation. MATERIALS AND METHODS: Network pharmacology and molecular docking were used to screen and validate candidate DL targets for RA treatment, respectively. A DBA/1 mouse rheumatoid arthritis model was induced with bovine type II collagen. Intragastric DL administration was followed by the calculation of the clinical arthritis index. A section of the ankle joint was excised and stained and the pathological changes in it were observed. Enzyme-linked immunosorbent assays (ELISA) and western blotting (WB) were used to clarify the mechanisms of DL in RA treatment. RESULTS: DL effectively attenuated the inflammation, mitigated the articular cartilage degradation, and bone erosion, and alleviated the inflammatory joints associated with RA. Network pharmacology screened six key targets of DL while molecular docking revealed that it docked well with its protein targets. The DL treatment group presented with significantly less ankle joint redness and swelling, a lower arthritis index scores and serum and bone marrow supernatant IL-6 levels, more complete ankle joint surfaces, and less synovial inflammation, cartilage degradation, and bone erosion than the collagen-induced arthritis (CIA) group. The DL treatment also substantially downregulated the Janus kinase (JAK)1, JAK3, matrix metalloproteinase (MMP)2, MMP9, and phospho-signal transducer and activator of transcription (p-STAT)3 proteins in the joints. CONCLUSIONS: To the best of our knowledge, the present work was the first to demonstrate that DL has significant anti-inflammatory efficacy and reduces cartilage degradation and bone erosion. It also demonstrated that the anti-RA effect of DL may be explained by its ability to inhibit joint inflammation and reduce articular cartilage degradation through the interleukin (IL)-6/JAK1,3/STAT3 axis and downregulate MMP2 and MMP9. Hence, DL might play a therapeutic role in a mouse RA model.

Deep learning based atomic defect detection framework for two-dimensional materials.

Defects to popular two-dimensional (2D) transition metal dichalcogenides (TMDs) seriously lower the efficiency of field-effect transistor (FET) and depress the development of 2D materials. These atomic defects are mainly identified and researched by scanning tunneling microscope (STM) because it can provide precise measurement without harming the samples. The long analysis time of STM for locating defects in images has been solved by combining feature detection with convolutional neural networks (CNN). However, the low signal-noise ratio, insufficient data, and a large amount of TMDs members make the automatic defect detection system hard to be applied. In this study, we propose a deep learning-based atomic defect detection framework (DL-ADD) to efficiently detect atomic defects in molybdenum disulfide (MoS2) and generalize the model for defect detection in other TMD materials. We design DL-ADD with data augmentation, color preprocessing, noise filtering, and a detection model to improve detection quality. The DL-ADD provides precise detection in MoS2 (F2-scores is 0.86 on average) and good generality to WS2 (F2-scores is 0.89 on average).

Five previously undescribed compounds from Ajuga lupulina Maxim. and their in vitro activities.

A new biflavone (philonotisflavone-3'''-methyl ether), three diterpenes (lupulin G, lupulin H, lupulin I), a new ecdysteroid (ajugasterone E), and four known compounds were isolated from the whole plant of Ajuga lupulina Maxim. Their structures were determined by spectroscopic analysis, including MS, NMR and ECD spectral data. Compounds 1-3 has DPPH radical scavenging ability, and compound 1 has stronger activity than vitamin C. Compounds 2, 3, 7 and 8 have potential cytotoxic activity against Hela, with IC50 values less than 20.0 µM. Abietane diterpenes 2, 3, 7 and 8 are also found to have NO inhibitory effects with IC50 values less than 40.0 µM.

Euphorfiatnoids A-I: Diterpenoids from the roots of Euphorbia fischeriana with cytotoxic effects.

Nine previously undescribed diterpenoids, euphorfiatnoids A-I, together with seven known diterpenoids, were isolated from the roots of wild Euphorbia fischeriana. Their structures were elucidated by the interpretation of HRESIMS, UV, and NMR data. Their configurations were determined by electronic circular dichroism (ECD) spectroscopy analysis and the structure of euphorfiatnoid A was confirmed by X-ray crystallography. To further understand the antitumor effects of E. fischeriana, we tested the cytotoxicity of these compounds against H460, HepG2, and MCF-7 cell lines in vitro using MTT assays. Euphorfiatnoid B exhibited the most promising inhibitory effect against H460 cells with an IC50 value of 9.97 µM. Euphorfiatnoid A and C also exhibited moderate cytotoxicity against HepG2 cells with IC50 values of 11.64 and 13.10 µM, respectively.

Molecular cloning and characterization of three phenylalanine ammonia-lyase genes from Schisandra chinensis.

Phenylalanine ammonia-lyase (PAL), which catalyzes the conversion from L-phenylalanine to trans-cinnamic acid, is a well-known key enzyme and a connecting step between primary and secondary metabolisms in the phenylpropanoid biosynthetic pathway of plants and microbes. Schisandra chinensis, a woody vine plant belonging to the family of Magnoliaceae, is a rich source of dibenzocyclooctadiene lignans exhibiting potent activity. However, the functional role of PAL in the biosynthesis of lignan is relatively limited, compared with those in lignin and flavonoids biosynthesis. Therefore, it is essential to clone and characterize the PAL genes from this valuable medicinal plant. In this study, molecular cloning and characterization of three PAL genes (ScPAL1-3) from S. chinensis was carried out. ScPALs were cloned using RACE PCR. The sequence analysis of the three ScPALs was carried out to give basic characteristics followed by docking analysis. In order to determine their catalytic activity, recombinant protein was obtained by heterologous expression in pCold-TF vector in Escherichia coli (BL21-DE3), followed by Ni-affinity purification. The catalytic product of the purified recombinant proteins was verified using RP-HPLC through comparing with standard compounds. The optimal temperature, pH value and effects of different metal ions were determined. Vmax, Kcat and Km values were determined under the optimal conditions. The expression of three ScPALs in different tissues was also determined. Our work provided essential information for the function of ScPALs.

[A new norsesquiterpenoid from Schisandra chinensis].

This study investigated the chemical components from the leaves and stems of Schisandra chinensis. Three norsesquiterpenoids were isolated from S. chinensis by various column chromatographies(silica gel, Sephadex LH-20, and MCI), reversed-phase medium-pressure preparative, and semi-preparative high-performance liquid chromatography(HPLC). Their structures were identified based on physicochemical properties, mass spectrometry(MS), nuclear magnetic resonance(NMR), ultraviolet(UV), and electro-nic circular dichroism(ECD) as(3R,4R,5R,6S,7E)-3,4,5,6-tetrahydroxy-7-megastigmen-9-one(1),(3S,5R,6R,7E)-3,5,6-trihydroxy-7-megastigmen-9-one(2), and(3S,4R,9R)-3,4,9-trihydroxymegastigman-5-ene(3). Compound 1 was a new compound, and its absolute configuration was determined by ECD. Compounds 2 and 3 were isolated from the Schisandra plant for the first time.

Four sesquiterpenoids from the vegetable raw material Schisandra chinensis (Turcz.) Baill.: leaves and stems.

Four sesquiterpenoids A-D (1-4) were isolated from the ethanol extracts of the leaves and stems from Schisandra chinensis. Their structures and absolute configurations were elucidated by a combination of NMR, MS and ECD. Compounds 1-4 (10 µM) exhibited moderate hepatoprotective activities against APAP-induced LO2 cell damage with increasing cell viability by 18%, 17%, 16%, and 19% compared to the model group (bicyclol, 26%) at 10 µM, respectively. All the compounds displayed no cytotoxic activity against five human cell lines, which also suggested the safety of leaves and stems of S. chinensis as an edible vegetable in a degree.

[Cloning of AhWRKY33 from Asarum heterotropoides and its genetic transformation in Arabidopsis].

The WRKY family genes, which play an important role in plant morphogenesis and stress response, were selected based on the data of the full-length transcriptome of Asarum heterotropoides. Using AtWRKY33, which regulates the synthesis of the camalexin in the model plant Arabidopsis to compare homologous genes in A. heterotropoides, primers were designed to amplify the open reading frame(ORF) fragment of AhWRKY33 gene by RT-PCR using total RNA of A. heterotropoides leaves as template. Real-time PCR results showed that there was a significant difference between the aerial part and the underground part of A. heterotropoides, the toxic aristolochic acid content is highly expressed in the leaves higher than the root. After verification, the WRKY33 gene of A. heterotropoides is ORF long 1 686 bp, encoding 561 amino acids.AhWRKY33 had two conserved WRKYGQK domains. According to the classical classification, it belongs to group Ⅰ WRKY transcription factor. A. heterotropoides WRKY33 had some homology with amino acids of other species. The study successfully constructed the plant eukaryotic expression vector PHG-AhWRKY33 and transformed Arabidopsis thaliana, the transgenic Arabidopsis was obtained by PCR detection and hygromycin resistant plate screening. It found that the germination of transgenic Arabidopsis seeds was accelerated and the stress resistance was increased. It laid a foundation for further analysis of WRKY transcription factor in the growth and development of A. heterotropoides and the synthesis of secondary metabolites.

Phytochemical and antitumor studies on Cynanchum mongolicum (Maxim.) Kom.

A chemical investigation of Cynanchum mongolicum (Maxim.) Kom. identified 8 compounds. On the basis of spectroscopic data, they were determined to be 3 alkaloids and 5 sinapoyl esters, among which were two previously undescribed compounds (1 and 2). The inhibitory effects of the isolated compounds against four human tumor cell lines were evaluated in vitro by MTT assays, which revealed moderate inhibitory effects with IC50 values < 50 mM, in particularly, three antofine analogues have showed significant antitumor activities with IC50 values < 0.1 mM, which was obviously better than the 5-fluorouracil and potential to be used as cancer drugs.