Unraveling the anti-primary dysmenorrhea mechanism of Ainsliaea fragrans Champ. extract by the integrative approach of network pharmacology and experimental verification.

BACKGROUND: The plant Ainsliaea fragrans Champ. (A. fragrans) named "Xingxiang Tuerfeng", is a traditional herb with a long history of therapeutic practice in southern China in the treatment of gynecological diseases. PURPOSE: The anti-inflammatory extract of Ainsliaea fragrans Champ. (AF-ext) exhibited anti-primary dysmenorrhea (PD) activity in oxytocin-induced mice. This study aimed to unravel the underlying mechanisms of AF-ext on PD by the integrative approach of network pharmacology and experimental verification. METHODS: First, the therapeutic targets of AF-ext are predicted using network pharmacology and molecular docking methods. Second, activity screening and immunoblotting methods were used for target validation. Then, the therapeutic effect of AF-ext on PD was evaluated using oxytocin-induced mice and uterine strips model. RESULTS: AF-p1, and AF-p2, the active ingredients of AF-ext, showed inhibitory effects on COX1/2 and EGFR, and all five active components showed antagonistic activity on TRPV1. AF-ext (25, 50, 100 mg/kg) could significantly reduce the number of writhing times and prolong writhing latencies in a dose-dependent manner. AF-ext inhibited spasmolytic activity in uterine strips induced by oxytocin and Ca2+ stimulation. AF-ext inhibited NF-κB/COX-2/PG pathway and activation of the NLRP3 inflammasome in PD mice. It significantly downregulated the PD-induced overexpression of p-p65/p65, p-IκBα, and COX-2 by inhibiting the NF-κB pathway. Moreover, the overexpression of NLRP3, p20/pro-Caspase 1, and p17/pro-IL-1ß was greatly downregulated. CONCLUSIONS: AF-ext demonstrated anti-inflammatory, analgesic, and spasmolytic activity in the treatment of PD. It inhibited the NF-κB/COX-2/PG pathway and NLRP3 inflammasome activation in PD mice with a multi-target approach.

Traditional uses, chemical composition, and pharmacological effects of diaphragma juglandis fructus: A review.

ETHNOPHARMACOLOGICAL RELEVANCE: In traditional Uyghur medicine, diaphragma juglandis fructus (DJF) has been conventionally used in treating insomnia and nourishing the kidneys. According to traditional Chinese medicine, DJF can boosts kidney and astringent essence, strengthen the spleen and kidney, exert diuretic effect, clear heat, stop eructation, and treat vomiting. AIM OF THE REVIEW: Research on DJF has increased gradually in recent years, but reviews of its traditional uses, chemical composition, and pharmacological activities are scarce. The purpose of this review is to analyze the traditional uses, chemical composition, and pharmacological activities of DJF and provide an overview of the findings for further research and development of DJF resources. MATERIALS AND METHODS: Data on DJF were obtained from different databases, including Scifinder, PubMed, Web of Science, Science Direct, Springer, Wiley, ACS, CNKI, Baidu Scholar, and Google Scholar; books; and Ph.D. and MSc theses. RESULTS: According to traditional Chinese medicine, DJF has astringent properties, inhibits bleeding and banding, strengthens the spleen and kidneys, acts as a sleeping aid by reducing anxiety, and relieves dysentery due to heat exposure. The components of DJF include flavonoids, phenolic acids, quinones, steroids, lignans, and volatile oils, which exhibit good antioxidant, antitumor, antidiabetic, antibacterial, anti-inflammatory, and sedative-hypnotic properties, and present therapeutic potential for kidney diseases. CONCLUSIONS: Based on its traditional use, chemical composition, and pharmacological activities, DJF is a promising source of natural medicine in the development of functional foods, drugs, and cosmetics.

Centella asiatica (L.) Urb. attenuates cardiac hypertrophy and improves heart function through multi-level mechanisms revealed by systems pharmacology.

ETHNOPHARMACOLOGICAL RELEVANCE: Cardiac hypertrophy (CH) is an incurable heart disease, contributing to an increased risk of heart failure due to the lack of safe and effective strategies. Therefore, searching for new approaches to treat CH is urgent. Centella asiatica (L.) Urb. (CA), a traditional food and medicinal natural plant, has been turned out to be effective in the treatment of cardiovascular disease, but its efficacy and potential mechanisms in alleviating CH have not yet been investigated. AIM OF STUDY: In this study, we aimed to elucidate the multi-level mechanisms underlying the effect of CA against CH. STUDY DESIGN AND METHODS: A systems pharmacology approach was employed to screen active ingredients, identify potential targets, construct visual networks and systematically investigate the pathways and mechanisms of CA for CH treatment. The cardiac therapeutic potential and mechanism of action of CA on CH were verified with in vivo and in vitro experiments. RESULTS: Firstly, we demonstrated the therapeutic effect of CA on CH and then screened 13 active compounds of CA according to the pharmacokinetic properties. Then, asiatic acid (AA) was identified as the major active molecule of CA for CH treatment. Afterwards, network and functional enrichment analyses showed that CA exerted cardioprotective effects by modulating multiple pathways mainly involved in anti-apoptotic, antioxidant and anti-inflammatory processes. Finally, in vivo, the therapeutic effects of AA and its action on the YAP/PI3K/AKT axis and NF-κB signaling pathway were validated using an isoproterenol-induced CH mouse model. In vitro, AA decreased ROS levels in hydrogen peroxide-treated HL-1 cells. CONCLUSION: Overall, the multi-level mechanisms of CA for CH treatment were demonstrated by systems pharmacology approach, which provides a paradigm for systematically deciphering the mechanisms of action of natural plants in the treatment of diseases and offers a new idea for the development of medicinal and food products.

Systems pharmacology reveals the multi-level synergetic mechanism of action of Ginkgo biloba L. leaves for cardiomyopathy treatment.

ETHNOPHARMACOLOGICAL RELEVANCE: Cardiomyopathy is a common cause of heart failure and may lead to increased risk of sudden cardiac death, lacking simple, safe and effective treatment strategies due to unclear pathogenesis. Ginkgo biloba L. leaves (GBLs), a traditional Chinese medicine (TCM), has been widely used in clinical medicine for improving blood circulation, and was demonstrated to be effective on cardiomyopathy in preclinical studies. However, because of the widely known holistic therapeutic philosophy via multi-target and multi-pathway effect for most TCMs, to explore its underlying molecular mechanisms of action (MoA) remains a great challenge. AIM OF STUDY: Decipher the underlying MoA of GBLs for cardiomyopathy treatment: Study design and methods: An integrated systems pharmacology framework was employed to screen potential active compounds, identify therapeutic targets, explore the action pathways and verify mechanisms of GBLs with in vitro experiments. RESULTS: We firstly confirmed the therapeutic effect of GBLs on cardiomyopathy and subsequently screened 27 active compounds from GBLs according to their pharmacokinetic properties. Then Probability Ensemble Approach was applied to identify the compound combinations that exert synergetic effect from GBLs. Network analysis and functional enrichment analysis demonstrated that these compounds exhibit synergistic therapeutic effect by acting on multiple targets and thereby regulating multiple pathways mainly involved in pro-survival, anti-apoptotic and anti-inflammatory processes. Finally, using a doxorubicin-induced myocardial injury model, therapeutic effect of ginkgolide A, ginkgolide B, isorhamnetin, as well as their synergistic effect on PI3K-AKT and NF-κB signaling pathways were validated in vitro. Importantly, we demonstrated that Ginkgo diterpene lactone meglumine injection (GDJ), an approved injection derived from GBLs, could be a promising agent for cardiomyopathy treatment. CONCLUSION: Collectively, the multi-level synergetic mechanism of GBLs on cardiomyopathy treatment was demonstrated with systems pharmacology approach, providing a paradigm for deciphering the complicated MoA of TCMs.

A chromosome-scale genome assembly of a diploid alfalfa, the progenitor of autotetraploid alfalfa.

Alfalfa (Medicago sativa L.) is one of the most important and widely cultivated forage crops. It is commonly used as a vegetable and medicinal herb because of its excellent nutritional quality and significant economic value. Based on Illumina, Nanopore and Hi-C data, we assembled a chromosome-scale assembly of Medicago sativa spp. caerulea (voucher PI464715), the direct diploid progenitor of autotetraploid alfalfa. The assembled genome comprises 793.2 Mb of genomic sequence and 47,202 annotated protein-coding genes. The contig N50 length is 3.86 Mb. This genome is almost twofold larger and contains more annotated protein-coding genes than that of its close relative, Medicago truncatula (420 Mb and 44,623 genes). The more expanded gene families compared with those in M. truncatula and the expansion of repetitive elements rather than whole-genome duplication (i.e., the two species share the ancestral Papilionoideae whole-genome duplication event) may have contributed to the large genome size of M. sativa spp. caerulea. Comparative and evolutionary analyses revealed that M. sativa spp. caerulea diverged from M. truncatula ~5.2 million years ago, and the chromosomal fissions and fusions detected between the two genomes occurred during the divergence of the two species. In addition, we identified 489 resistance (R) genes and 82 and 85 candidate genes involved in the lignin and cellulose biosynthesis pathways, respectively. The near-complete and accurate diploid alfalfa reference genome obtained herein serves as an important complement to the recently assembled autotetraploid alfalfa genome and will provide valuable genomic resources for investigating the genomic architecture of autotetraploid alfalfa as well as for improving breeding strategies in alfalfa.

Translational pharmacokinetic-pharmacodynamic analysis in the pharmaceutical industry: an IQ Consortium PK-PD Discussion Group perspective.

With inadequate efficacy being the primary cause for the attrition of drug candidates in clinical development, the need to better predict clinical efficacy earlier in the drug development process has increased in importance in the pharmaceutical industry. Here, we review current applications of translational pharmacokinetic-pharmacodynamic (PK-PD) modeling of preclinical data in the pharmaceutical industry, including best practices. Preclinical translational PK-PD modeling has been used in many therapeutic areas and has been impactful to drug development. The role of preclinical translational PK-PD modeling in drug discovery and development will continue to evolve and broaden, given that its broad implementation in the pharmaceutical industry is relatively recent and many opportunities still exist for its further application.

High Colonization Possibility of Some Species of Weeds in Suaeda salsa Community: From an Ecological Stoichiometry Perspective.

Suaeda salsa community is a vegetation type in saline-alkali areas. Weed invasion and colonization in S. salsa communities lead to fragmentationsof S. salsa communities. The colonization of invaded weeds in S. salsa communities is related to community succession of saline-alkali zones. The fragmented S. salsa community may be restored if the mechanism of invaded weed colonization in S. salsa communities is clearly elucidated. Thus, we studied the ecological stoichiometric characteristics of soils and plants in a salt marsh to explain the high colonization possibility of invaded weeds in S. salsa communities. In October 2014, soils and plants were collected from Dongfeng Salt Marsh, Jiaozhou Bay, Shandong Province, China. The ratio of Ex-N/Ex-P in soil was less than 13, which suggests a relative nitrogen limitation for the primary production in the zone. The minimum phosphorus content in plants was higher than 1 mg g-1, whereas the maximum nitrogen content in plants was less than 13 mg g-1. These results imply that phosphorus was abundant, whereas nitrogen was deficient in the area. The plants in the salt marsh may be limited by nitrogen. Given the relatively lower nitrogen contents in Cyperus glomeratus, Echinochloa crusgalli, and Aster subulatus than that in S. salsa, these three species exhibited higher competitiveness than S. salsa did when nitrogen was limited in primary production. These weed species may colonize highly in S. salsa communities. Moreover, nitrogen fertilization might be effective to maintain S. salsa community in Dongfeng Salt Marsh, whereas its effects on controlling weeds colonization in S. salsa communities need more studies to verify.

Ocean acidification: One potential driver of phosphorus eutrophication.

Harmful algal blooms which may be limited by phosphorus outbreak increases currently and ocean acidification worsens presently, which implies that ocean acidification might lead to phosphorus eutrophication. To verify the hypothesis, oxic sediments were exposed to seawater with different pH 30days. If pH was 8.1 and 7.7, the total phosphorus (TP) content in sediments was 1.52±0.50 and 1.29±0.40mg/g. The inorganic phosphorus (IP) content in sediments exposed to seawater with pH8.1 and 7.7 was 1.39±0.10 and 1.06±0.20mg/g, respectively. The exchangeable phosphorus (Ex-P) content in sediments was 4.40±0.45 and 2.82±0.15µg/g, if seawater pH was 8.1 and 7.7. Ex-P and IP contents in oxic sediments were reduced by ocean acidification significantly (p<5%). The reduced phosphorus in sediments diffused into water, which implied that ocean acidification was one potential facilitator of phosphorus eutrophication in oxic conditions.

Tetrandrine is a potent cell autophagy agonist via activated intracellular reactive oxygen species.

BACKGROUND: Autophagy is an evolutionarily conserved cellular process that involves the lysosomal degradation of proteins and organelles and the recycling of cellular components to ensure cellular survival under external or internal stress. Numerous data has indicated that autophagy can be successfully targeted for the treatment of multiple cancers. We have previously demonstrated that tetrandrine, a bisbenzylisoquinoline alkaloid isolated from the broadly used Chinese medicinal herb Stephaniae tetrandrae, exhibits potent antitumor effects when used either alone or in combination with other drugs. RESULTS: In the present study, we showed that tetrandrine is a broad-spectrum potent autophagy agonist. Although low-dose tetrandrine treatment does not affect cell viability, it can potently induce autophagy in a variety of cell lines, including cancerous cells and nontumorigenic cells. The autophagy inhibitors 3-methyladenine (3-MA) and chloroquine (CQ), effectively blocked tetrandrine-induced autophagy. Moreover, tetrandrine significantly triggered the induction of mitophagy. The underlying mechanisms are associated with the tetrandrine-induced production of intracellular reactive oxygen species (ROS), which plays a critical role in tetrandrine-induced autophagy. CONCLUSIONS: Here, we report that tetrandrine is a potent cell autophagy agonist and may have a wide range of applications in the fields of antitumor therapy and basic scientific research.

Microbial side-chain cleavage of phytosterols by mycobacteria in vegetable oil/aqueous two-phase system.

Microbial side-chain cleavage of natural sterols to 4-androstene-3,17-dione (AD) and 1,4-androstadiene-3,17-dione (ADD) by Mycobacteria has received much attention in pharmaceutical industry, while low yield of the reaction owing to the strong hydrophobicity of sterols is a tough problem to be solved urgently. Eight kinds of vegetable oils, i.e., sunflower, peanut, corn, olive, linseed, walnut, grape seed, and rice oil, were used to construct oil/aqueous biphasic systems in the biotransformation of phytosterols by Mycobacterium sp. MB 3683 cells. The results indicated that vegetable oils are suitable for phytosterol biotransformation. Specially, the yield of AD carried out in sunflower biphasic system (phase ratio of 1:9, oil to aqueous) was greatly increased to 84.8 % with 10 g/L feeding concentration after 120-h transformation at 30 °C and 200 rpm. Distribution coefficients of AD in different oil/aqueous systems were also determined. Because vegetable oils are of low cost and because of their eco-friendly characters, there is a great potential for the application of oil/aqueous two-phase systems in bacteria whole cell biocatalysis.

The Arabidopsis kinesin gene AtKin-1 plays a role in the nuclear division process during megagametogenesis.

KEY MESSAGE: Atkin - 1 , the only Kinesin-1 member of Arabidopsis thaliana , plays a role during female gametogenesis through regulation of nuclear division cycles. Kinesins are microtubule-dependent motor proteins found in eukaryotic organisms. They constitute a superfamily that can be further classified into at least 14 families. In the Kinesin-1 family, members from animal and fungi play roles in long-distance transport of organelles and vesicles. Although Kinesin-1-like sequences have been identified in higher plants, little is known about their function in plant cells, other than in a recently identified Kinesin-1-like protein in a rice pollen semi-sterile mutant. In this study, the gene encoding the only Kinesin-1 member in Arabidopsis, AtKin-1 was found to be specifically expressed in ovules and anthers. AtKin-1 loss-of-function mutants showed substantially aborted ovules in siliques, and this finding was supported by complementation testing. Reciprocal crossing between mutant and wild-type plants indicated that a defect in AtKin-1 results in partially aborted megagametophytes, with no observable effects on pollen fertility. Further observation of ovule development in the mutant pistils indicated that the enlargement of the megaspore was blocked and nuclear division arrested at the one-nucleate stage during embryo sac formation. Our data suggest that AtKin-1 plays a role in the nuclear division cycles during megagametogenesis.