Effect and mechanism of Prunella vulgaris L. extract on alleviating lipopolysaccharide-induced acute mastitis in protecting the blood-milk barrier and reducing inflammation.

ETHNOPHARMACOLOGICAL RELEVANCE: According to ancient literature, Prunella vulgaris L. (P vulgaris) alleviates mastitis and has been used in China for many years; however, there are no relevant reports that confirm this or the mechanism of its efficacy. AIM OF THE STUDY: To explore the anti-acute mastitis effect and potential mechanism of P vulgaris extract. MATERIALS AND METHODS: First, the active ingredients and targets of P vulgaris against mastitis were predicted using network pharmacology. Next, the relevant active ingredients were enriched using macroporous resins and verified using UV and UPLC-Q-TOF-MS/MS. Lastly, a mouse model of acute mastitis was established by injecting lipopolysaccharides into the mammary gland and administering P vulgaris extract by oral gavage. The pathological changes in mammary tissue were observed by HE staining. Serum and tissue inflammatory factors were measured by ELISA method. MPO activity in mammary tissue was measured using colorimetry and MPO expression was detected by immunohistochemistry. The expression of tight junction proteins (ZO-1, claudin-3, and occludin) in mammary tissue was detected by immunofluorescence and Western blot. iNOS and COX-2 in mammary tissue were detected by Western blot. MAPK pathway and NF-κB pathway related proteins were also detected by Western blot. RESULTS: Network pharmacology predicted that phenolic acids and flavonoids in P vulgaris had anti-mastitis effects. The contents of total flavonoids and total phenolic acids in P vulgaris extract were 64.5% and 29.4%, respectively. UPLC-Q-TOF-MS/MS confirmed that P vulgaris extract contained phenolic acids and flavonoids. The results of animal experiments showed that P vulgaris extract reduced lipopolysaccharide-induced inflammatory edema, inflammatory cell infiltration, and interstitial congestion of mammary tissue. It also reduced the levels of serum and tissue inflammatory factors TNF-α, IL-6, and IL-1ß, and inhibited the activation of MPO. Furthermore, it downregulated the expression of MAPK and NF-κB pathway-related proteins. The expressions of ZO-1, occludin, and claudin-3 in mammary gland tissues were upregulated. CONCLUSIONS: P vulgaris extract can maintain the integrity of mammary connective tissue and reduce its inflammatory response to prevent acute mastitis. Its mechanism probably involves regulating NF-κB and MAPK pathways.

Recent advances in medicinal and edible homologous plant polysaccharides: Preparation, structure and prevention and treatment of diabetes.

Medicinal and edible homologs (MEHs) can be used in medicine and food. The National Health Commission announced that a total of 103 kinds of medicinal and edible homologous plants (MEHPs) would be available by were available in 2023. Diabetes mellitus (DM) has become the third most common chronic metabolic disease that seriously threatens human health worldwide. Polysaccharides, the main component isolated from MEHPs, have significant antidiabetic effects with few side effects. Based on a literature search, this paper summarizes the preparation methods, structural characterization, and antidiabetic functions and mechanisms of MEHPs polysaccharides (MEHPPs). Specifically, MEHPPs mainly regulate PI3K/Akt, AMPK, cAMP/PKA, Nrf2/Keap1, NF-κB, MAPK and other signaling pathways to promote insulin secretion and release, improve glycolipid metabolism, inhibit the inflammatory response, decrease oxidative stress and regulate intestinal flora. Among them, 16 kinds of MEHPPs were found to have obvious anti-diabetic effects. This article reviews the prevention and treatment of diabetes and its complications by MEHPPs and provides a basis for the development of safe and effective MEHPP-derived health products and new drugs to prevent and treat diabetes.

Nuezhenoside G13 from Osmanthus fragrans fruit ameliorates Concanavalin A-induced autoimmune hepatitis by regulating the NF-κB/MAPK pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Osmanthus fragrans fruit (OFF) exhibits hepatoprotective function, and it is consumed as food and used in traditional medicine in China. Nuezhenoside G13 (G13) is present in the highest levels in OFF. Autoimmune hepatitis (AIH) is a manifestation of liver disease and seriously endangers health. However, it remains unclear whether G13 affects AIH. AIM OF THE STUDY: To clarify the effect of G13 on AIH and its exact underlying mechanism from a new perspective. MATERIALS AND METHODS: We used a Concanavalin A-induced AIH mouse model and lipopolysaccharide-treated Raw264.7 cells to quantify serum biochemical indicators and confirm whether G13 exhibited protective effects in the AIH mice. Furthermore, we evaluated the effect of G13 via hematoxylin and eosin and immunohistochemical staining. We used enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction to quantify the inflammatory factors. We confirmed that G13 inhibited apoptosis via terminal deoxynucleotidyl transferase dUTP nick end labeling staining. Molecular docking, immunofluorescence, and western blotting experiments of G13 and key proteins of the NF-κB/MAPK pathway revealed that G13 alleviated inflammation. In addition, Cell Counting Kit-8, ELISA, NO detection, and western blotting assays were performed. Finally, we used an inhibitor of the p38 MAPK to verify that G13 reduced inflammation through the NF-κB/MAPK pathway in Raw264.7 cells. RESULTS: The in vivo experiments revealed that G13 improved oxidative stress and apoptosis. In addition, G13 decreased the expression levels of CD4+, CD8+, F4/80+, and Ly6G and the secretion of inflammatory factors. Interestingly, G13 reduced the phosphorylation levels of IκBα, NF-κB, JNK, ERK1/2, and p38. Additionally, the in vitro experiments revealed that G13 alleviated inflammation through the NF-κB/MAPK pathway in lipopolysaccharide-treated Raw264.7 cells. Furthermore, molecular docking demonstrated that the binding fraction of G13 with these proteins was high. CONCLUSION: G13 suppressed oxidative stress, apoptosis, and inflammation in a Concanavalin A-induced AIH mouse model. Furthermore, G13 exerted its effect through the NF-κB/MAPK pathway.

A chromosome-scale Rhubarb (Rheum tanguticum) genome assembly provides insights into the evolution of anthraquinone biosynthesis.

Rhubarb is the collective name for various perennial plants from the genus Rheum L. and the Polygonaceae family. They are one of the most ancient, commonly used, and important herbs in traditional Chinese medicine. Rhubarb is a major source of anthraquinones, but how they are synthesized remains largely unknown. Here, we generate a genome sequence assembly of one important medicinal rhubarb R. tanguticum at the chromosome level, with 2.76 Gb assembled into 11 chromosomes. The genome is shaped by two recent whole-genome duplication events and recent bursts of retrotransposons. Metabolic analyses show that the major anthraquinones are mainly synthesized in its roots. Transcriptomic analysis reveals a co-expression module with a high correlation to anthraquinone biosynthesis that includes key chalcone synthase genes. One CHS, four CYP450 and two BGL genes involved in secondary metabolism show significantly upregulated expression levels in roots compared with other tissues and clustered in the co-expression module, which implies that they may also act as candidate genes for anthraquinone biosynthesis. This study provides valuable insights into the genetic bases of anthraquinone biosynthesis that will facilitate improved breeding practices and agronomic properties for rhubarb in the future.

Polyethylene glycol-modified nanoscale conjugated polymer for the photothermal therapy of lung cancer.

Killing tumor cells efficiently with photothermal therapy remains a huge challenge. In this study, we successfully prepared a novel polymer with photothermal conversion capability via a condensation reaction, and then subjected it to Polyethylene glycol (PEG) modification and ultrasonic nanocrystalline treatment to make it suitable forin vivophotothermal therapy applications. The conjugated polymer demonstrated good biocompatibility and photothermal conversion ability and was shown in cell experiments to be effective in killing tumor cells after laser irradiation. In addition, the conjugated polymer-based photothermal therapy, guided by photoacoustic real-time imaging and mediated by laser irradiation, of a tumor-bearing mouse model could effectively inhibit the growth of tumor tissue and demonstrated goodin vivobiosafety. Thus, photothermal therapy based on the conjugated polymer synthesized in this study provides a new idea and strategy for the treatment of lung cancer.

Gypensapogenin I Suppresses Cell Proliferation in Triple-Negative Breast Cancer Via Triggering the Closure of AKT/GSK3ß/ß-Catenin and Notch-1 Signaling Pathways.

Jiaogulan (Gynostemma pentaphyllum) tea is a functional food that is commercially available worldwide. Gypensapogenin I (Gyp I), which is a natural damarane-type saponin, was obtained from the hydrolysates of total gypenosides. The present research was performed to investigate the potential antiproliferation effect of Gyp I in MDA-MB-231 cells and the underlying mechanisms. Here, we found that Gyp I attenuated survival, inhibited proliferation, and induced apoptosis in MDA-MB-231 cells. Target prediction by binding molecule docking and western blot assays confirmed the mechanism by which Gyp I inhibited the proliferation of breast cancer cells via the AKT/GSK3ß/ß-catenin signaling pathway. We also showed that Gyp I exhibited superior in vivo efficacy that was dose dependent. Tumor tissue transcriptome analysis indicated that Gyp I could decrease the expression levels of NOTCH1 and HES1, which was in contrast to the effect on MAML and NUMBL, indicating that our compound hindered the activation of the Notch-1 signaling pathway. In summary, we report for the first time that Gyp I shows excellent anti-breast cancer activity in vivo and in vitro and that its pathway of action is related to the AKT/GSK3ß/ß-catenin and Notch-1 signaling pathways. Therefore, Jiaogulan tea can not only be used as a health food but also possesses the possibility to treat triple-negative breast cancer (TNBC).

Effectiveness and Safety of Aromatherapy in Managing Behavioral and Psychological Symptoms of Dementia: A Mixed-Methods Systematic Review.

INTRODUCTION: Behavioral and psychological symptoms of dementia (BPSD) is the most prominent and distressing manifestation for older persons with dementia (PWD) and caregivers. Aromatherapy has demonstrated its effectiveness in managing BPSD in various studies. However, previous studies and systematic reviews have obtained inconsistent findings, and a review of qualitative studies is yet to be conducted. METHOD: A mixed-methods systematic review with a convergent segregated approach was performed to evaluate the effectiveness of aromatherapy in improving the BPSD and quality of life (QoL) of PWD and in relieving the distress and burden of caregivers, as well as its safety for PWD. Both published and unpublished quantitative and qualitative studies written in English and Chinese between January 1996 and December 2020 were retrieved from 28 databases, including MEDLINE, EMBASE, and Web of Science, based on the prespecified criteria. The methodological quality was assessed by using critical appraisal tools from the Joanna Briggs Institute. Quantitative synthesis, qualitative synthesis, and integration of quantitative and qualitative evidence were performed. RESULTS: A total of 12 randomized controlled trials, 10 quasi-experimental studies, and 2 qualitative studies were included in the review. Some inconsistent findings regarding the effectiveness of aromatherapy in reducing the severity of BPSD were observed. Some studies reported that aromatherapy significantly improved the QoL of PWD and relieved the distress and burden of caregivers, promoted a positive experience among caregivers, and had very low adverse effects on PWD (with aromatherapy inhalation reporting no adverse effects). CONCLUSION: Aromatherapy, especially in the inhalation approach, could be a potentially safe and effective strategy for managing BPSD. However, more structuralized and comparable studies with sufficient sample size, adherence monitoring, and sound theoretical basis could be conducted to obtain conclusive findings.

Covalent Organic Framework for Improving Near-Infrared Light Induced Fluorescence Imaging through Two-Photon Induction.

Fluorescent materials exhibiting two-photon induction (TPI) are used for nonlinear optics, bioimaging, and phototherapy. Polymerizations of molecular chromophores to form π-conjugated structures were hindered by the lack of long-range ordering in the structure and strong π-π stacking between the chromophores. Reported here is the rational design of a benzothiadiazole-based covalent organic framework (COF) for promoting TPI and obtaining efficient two-photon induced fluorescence emissions. Characterization and spectroscopic data revealed that the enhancement in TPI performance is attributed to the donor-π-acceptor-π-donor configuration and regular intervals of the chromophores, the large π-conjugation domain, and the long-range order of COF crystals. The crystalline structure of TPI-COF attenuates the π-π stacking interactions between the layers, and overcomes aggregation-caused emission quenching of the chromophores for improving near-infrared two-photon induced fluorescence imaging.

Evaluation of selected traditional Chinese medical extracts for bone mineral density maintenance: A mechanistic study.

OBJECTIVE: To investigate the development of a minimal traditional Chinese medicine (TCM) formula using selected TCM ingredients and evaluating their biological activity with bone-specific in vitro tests. Finally, determining if the minimal formula can maintain bone mineral density (BMD) in a low bone mass (LBM)/osteoporosis (OP) model system. METHODS AND RESULTS: Sixteen different TCM plant extracts were tested for estrogenic, osteogenic and osteoclastic activities. Despite robust activation of the full-length estrogen receptors α and ß by Psoralea corylifolia and Epimedium brevicornu, these extracts do not activate the isolated estrogen ligand binding domains (LBD) of either ERα or ERß; estrogen (17-ß estradiol) fully activates the LBD of ERα and ERß. E. brevicornu and Drynaria fortunei extracts activated cyclic AMP response elements (CRE) individually and when combined these ingredients stimulated the production of osteoblastic markers Runx2 and Bmp4 in MC3T3-E1 cells. E. brevicornu, Salvia miltiorrhiza, and Astragalus onobrychis extracts inhibited the Il-1ß mediated activation of NF-κß and an E. brevicornu/D. fortunei combination inhibited the development of osteoclasts from precursor cells. Further, a minimal formula containing the E. brevicornu/D. fortunei combination with or without a third ingredient (S. miltiorrhiza, Angelica sinensis, or Lycium barbarum) maintained bone mineral density (BMD) similar to an estradiol-treated control group in the ovariectomized rat; a model LBM/OP system. CONCLUSION: A minimal formula consisting of TCM plant extracts that activate CRE and inhibit of NF-κß activation, but do not behave like estrogen, maintain BMD in a LBM/OP model system.

Prevention Effects and Possible Molecular Mechanism of Mulberry Leaf Extract and its Formulation on Rats with Insulin-Insensitivity.

For centuries, mulberry leaf has been used in traditional Chinese medicine for the treatment of diabetes. This study aims to test the prevention effects of a proprietary mulberry leaf extract (MLE) and a formula consisting of MLE, fenugreek seed extract, and cinnamon cassia extract (MLEF) on insulin resistance development in animals. MLE was refined to contain 5% 1-deoxynojirimycin by weight. MLEF was formulated by mixing MLE with cinnamon cassia extract and fenugreek seed extract at a 6:5:3 ratio (by weight). First, the acute toxicity effects of MLE on ICR mice were examined at 5 g/kg BW dose. Second, two groups of normal rats were administrated with water or 150 mg/kg BW MLE per day for 29 days to evaluate MLE's effect on normal animals. Third, to examine the effects of MLE and MLEF on model animals, sixty SD rats were divided into five groups, namely, (1) normal, (2) model, (3) high-dose MLE (75 mg/kg BW) treatment; (4) low-dose MLE (15 mg/kg BW) treatment; and (5) MLEF (35 mg/kg BW) treatment. On the second week, rats in groups (2)-(5) were switched to high-energy diet for three weeks. Afterward, the rats were injected (ip) with a single dose of 105 mg/kg BW alloxan. After four more days, fasting blood glucose, post-prandial blood glucose, serum insulin, cholesterol, and triglyceride levels were measured. Last, liver lysates from animals were screened with 650 antibodies for changes in the expression or phosphorylation levels of signaling proteins. The results were further validated by Western blot analysis. We found that the maximum tolerance dose of MLE was greater than 5 g/kg in mice. The MLE at a 150 mg/kg BW dose showed no effect on fast blood glucose levels in normal rats. The MLE at a 75 mg/kg BW dose and MLEF at a 35 mg/kg BW dose, significantly (p < 0.05) reduced fast blood glucose levels in rats with impaired glucose and lipid metabolism. In total, 34 proteins with significant changes in expression and phosphorylation levels were identified. The changes of JNK, IRS1, and PDK1 were confirmed by western blot analysis. In conclusion, this study demonstrated the potential protective effects of MLE and MLEF against hyperglycemia induced by high-energy diet and toxic chemicals in rats for the first time. The most likely mechanism is the promotion of IRS1 phosphorylation, which leads to insulin sensitivity restoration.

Aqueous Extract of Chrysanthemum morifolium ( Jú Hua) Enhances the Antimelanogenic and Antioxidative Activities of the Mixture of Soy Peptide and Collagen Peptide.

The possible synergistic effect between the aqueous extract of Chrysanthemum morifolium ( Jú Hua) (AECM) and the peptide mixture (PM) containing soy peptide and collagen peptide was investigated in an ultraviolet (UV) irradiation-induced skin damage mouse model. The irradiated mice were treated with the PM or PM + AECM (containing PM and AECM), respectively. Both PM and PM + AECM groups displayed an apparent photoprotective effect on the UV-irradiated skin damage of mice. Histological evaluation demonstrated that the epidermal hyperplasia and melanocytes in the basal epidermal layer of the UV-irradiated skin in mice decreased when treated with either PM or PM + AECM. Further study showed that soy peptide, collagen peptide, and AECM also inhibited the activities of mushroom tyrosinase with IC50 values of 82.3, 28.2, and 1.6 µg/ml, respectively. Additionally, PM + AECM reduced melanogenesis by 46.2% at the concentration of 10 mg/ml in B16 mouse melanoma cells. Meanwhile, the UV-induced increase of antioxidative indicators, including glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and malondialdehyde (MDA), was reduced significantly after treatment with 1.83 g/kg/dbw of PM + AECM. This evidence supported the synergistic antioxidative effect of AECM with PM. These results demonstrated that oral intake of PM and AECM had synergistic antimelanogenic and antioxidative effects in UV-irradiated mice.

Two new sesquiterpenes from Acorus calamus.

A novel cadinane sesquiterpene with a propan-2-ylidene, named (1R,4R,6S,10R)-1-hydroxy-7(11)-cadinen-5,8-dione (1), and a new sesquiterpene, named (2R,6R,7S,9S)-1(10),4-cadinadiene-2,9-diol (2), were isolated from the ethanol extract of the rhizome of Acorus calamus L. Their structures were elucidated on the basis of spectroscopic data including NMR spectroscopy and X-ray crystallography. The absolute configurations of compounds 1 and 2 were confirmed by comparison of their experimental optical rotations with those obtained from theoretical calculations at the B3LYP/6-311+G (d) level of theory.

Resonance light-scattering spectrometric study of interaction between enzyme and MPA-modified CdTe nanoparticles.

This paper described a novel assay of enzyme based on the measurement of enhanced resonance light-scattering (RLS) signals resulting from the electrostatic and coordination interaction of functionalized CdTe nanoparticles with enzyme. The CdTe nanoparticles which were modified with 3-mercaptocarboxylic acid (MPA) have abundant carboxylic groups (COOH). So the nanoparticles are water-soluble, stable and biocompatible. At pH 8.3 phosphate buffered saline (PBS), the RLS signals of functionalized nano-CdTe are greatly enhanced by bromelain and papain in the region of 220-800 nm characterized by the peak around 318-314 nm, respectively. The optimization conditions of the reaction were also examined and selected. Under the selected conditions, the enhanced RLS intensity is linearly proportional to the concentration of bromelain and papain. The liner range is (0.09-0.9) x 10(-6)mol/L for bromelain and (0.048-0.702) x 10(-6)mol/L for papain. The influences of some foreign substances were also examined. This method can be applied to the determination of enzyme.

Bioactive secolignans from Peperomia dindygulensis.

Thirteen secolignans, including eight new ones (1-8), were isolated from the EtOAc extract of Peperomia dindygulensis. The structures were mainly elucidated by 1D and 2D NMR and MS experiments, the relative configurations were determined by NOE correlations, and the absolute configurations were established by the optical rotations and CD spectra. Cytotoxicity and MDR (multidrug resistance) reversal activity of the isolated compounds were examined. Compounds 6 and 7, peperomins B (10) and E (12), showed moderate to strong growth inhibitory activity against a malignant lung tumor cell (VA-13) with IC(50) values of 15.2, 13.5, 13.9, and 1.93 microM, respectively, and also inhibited the growth of a normal lung fibroblast cell (WI-38) at the same levels. Compound 7 and peperomin E (12) exhibited inhibitory activity against a liver tumor cell (HepG2) with IC(50) values of 22.3 and 12.1 microM. Compounds 5 and 7 and peperomins A, B, C, and E (9-12) enhanced calcein accumulation in MDR 2780 cells at 25 microg/mL. Compounds 2, 3, 7, and peperomin E (12) showed inhibitory activity on induction of the intercellular adhesion molecule-1 (ICAM-1).