The stems of Ephedra sinica and the fruits of Terminalia chebula are combined using in traditional Mongolian medicine formula “Gurigumu-7” for liver diseases. E. sinica stems contains ephedrine with ...broncho-dilatory activity. However, ephedrine can pass through the blood-brain barrier (BBB) and excite the central nervous system (CNS) to cause insomnia and restlessness.
The present study was to investigate the structures and bioactivities of new compounds formed in vivo after co-administration of E. sinica stems and T. chebula fruits.
Pharmacokinetic investigation was carried out in rats. A parallel artificial membrane permeability measurement system was used to determine BBB permeability. Ex vivo experiments using tracheal rings of guinea pig was performed to examine the tracheal relaxation effect. In vivo hepatoprotective tests were carried out in Tg (fabp10a: dsRed) liver transgenic zebrafish. The fluorescent probe, 2,7-dichlorodihydrofluorescein diacetate, was used to measure reactive oxygen species, and UHPLC-MS was used to determine glutathione concentrations after derivatization with N-ethylmaleimide.
New ephedrine derivatives (1 and 2) formed in vivo and reached their maximum serum concentrations at 0.5 h after administration of the two herbal drugs. Compounds 1 and 2 showed lower BBB permeability than ephedrine, suggesting that they have less adverse effects on the CNS. Compounds 1 and 2 relaxed the tracheal rings and had strong hepatoprotective effect on transgenic zebrafish with liver specific expression of RFP. Compounds 1 and 2 significantly reduced the level of reactive oxygen species while increasing that of glutathione in thioacetamide-treated zebrafish, which might be the hepatoprotective mechanism.
These results provided evidences that the chemical constituents in various herbal drugs in a medicinal formula can interact to generate new compounds with fewer side effects and increased or additive bioactivity.
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Abstract
Hypotrichs are among the most complex and highly differentiated ciliate lineages, which are distributed in diverse habitats. Members of the family Deviatidae Foissner, 2016 are commonly ...found in freshwater or low-salinity habitats, as well as soils. During a study of freshwater wetland diversity in China, a novel deviatid ciliate Heterodeviata sinica gen. nov. et sp. nov. and a distinct population of Deviata multilineae Zhang et al., 2022 were investigated based on morphology, morphogenesis, and 18S rRNA gene sequences. The new genus Heterodeviata gen. nov. can be characterized by having two or more left and right marginal rows each, one frontoventral cirral row, the presence of caudal cirri, and two dorsal kineties, one of them being a dorsomarginal kinety. Phylogenetic analyses reveal that Heterodeviata is nested in a well-supported clade with Deviata Eigner, 1995, Perisincirra Jankowski, 1978 and Pseudosincirra Gao et al., 2021, which suggests that the family Deviatidae is monophyletic. The close relationship among the Deviatidae genera were supported after extensive morphological and morphogenetic investigations, as well as molecular phylogeny presented here.
Though generally a mild affliction, allergic rhinitis (AR) is very common and causes considerable discomfort. Ephedra sinica polysaccharide is a candidate cost-effective therapy to relieve AR ...symptoms.
We explore the molecular mechanism of pure polysaccharide ESP-B4 action in AR.
RPMI2650 cells were treated with lipopolysaccharide to induce an in vitro sensitization model, and extracellular vesicles (EVs) were isolated. A rat model of AR was established using ovalbumin as the allergen and was treated with Ephedra sinica polysaccharide to observe changes in rhinitis symptoms, nasal mucosa histopathology and molecular pathology. ESP-B4-treated sensitized cells were adopted in vitro to verify effect of Ephedra sinica polysaccharide on miR-146a-5p expression in RPMI2650 cell-derived EVs and helper T cell differentiation.
miR-146a-5p inhibited Smad3, impeded the Smad3/GATA-3 interaction, upregulated IFN-γ expression, and promoted CD4+T cell Th1 differentiation. Treatment with ESP-B4 relieved AR in rats, and elevated miR-146a-5p in the EVs from the nasal epithelial cells, apparently in relation to effects on helper T cell Th1/Th2 equilibrium.
Overall, ESP-B4 can promote miR-146a-5p secretion, affect the Th1/Th2 balance of helper T cells, and relieve AR symptoms through Smad3/GATA-3 interaction, thus presenting a potential strategy for AR treatment.
Dried terrestrial stems of Ephedra sinica are known as ‘Ephedra herb.’ The pharmacological effects are mainly related to two major ingredients, (−)-ephedrine and (+)-pseudoephedrine (total alkaloids ...which are defined in Japanese Pharmacopoeia, TA). In this study, in order to aid in cultivation and breeding, the stability of TA content and stem dry weight of 46 E. sinica genets was evaluated from the first year of transplantation to the sixth year. TA content and composition ratio of these genets were stable after the second year, and dry weight was stable after the fourth year. These traits showed high inter-genet variability but low annual variability for each genet. Additionally, rank correlation coefficients of each trait among the genets were high. There was no significant correlation between these traits. Furthermore, to assess the reproducibility of these traits in clones, we evaluated TA content and dry weight of three clonal lines with high TA contents. TA content and composition ratio of the clonal lines were also stable after the second year of transplantation, and dry weight of the clonal lines was also stable after the fourth year. Moreover, TA content and composition ratio in each clonal line were comparable with those of each original genet after the second year. These results suggested that ephedrine alkaloids content and dry weight of E. sinica plants are stable, and that these traits are highly reproducible in clones. Therefore, selection breeding of E. sinica using vegetative propagation can be effective for high and stable quality of Ephedra herb.
To study the chemical constituents of the roots of Ephedra sinica Stapf.
The roots of E. sinica were extracted with 90% ethanol and isolated and purified by column chromatography on silica gel, ...Sephadex LH 20, and semipreparative RP-HPLC. All the compounds were identified based on spectral analysis (including MS, 1H-NMR and 13C-NMR, and 2D NMR) and compared with the reported literature. Their cytotoxicities were evaluated against three human cancer cell lines.
Two compounds were isolated from the roots of E. sinica and identified as (E)-eicosyl 3,4-dihydroxy-5-methoxycinnamate (1) and (E)-hexadecyl-ferulate (2).
Compound 1 was a new phenol named ephedrate A, and compound 2 was newly isolated from this plant. These two compounds showed no obvious cytotoxic activity, and their medicinal activities need to be studied further.
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•Glue-free biocomposites produced via hot pressing of artificial shrub.•Biocomposites show excellent hydrophobicity and high stability towards strong chemical.•Cell wall collapses ...enhanced mechanical bonding of fibrous components.•Depolymerization and repolymerization of lignin enhanced hydrogen bonding components.•Biocomposites obtained can potentially replace the composites prepared using adhesives.
There are abundant shrub species resources in the world, but their natural defects such as short growth cycle, loose material, easy cracking plus deformation, and low strength have seriously affected their scope of application. With the ever-decreasing of natural forest resources, the research on quality improvement and efficiency enhancement of small-diameter timber of artificial shrub represents a promising effort in global continual advancement. In this study, an innovative and environmentally-friendly approach was developed to produce high-quality biocomposites from Buxus sinica. Pretreatment of Buxus sinica in an alkaline sodium sulfite solution had degraded part of the amorphous structure from the cell walls. The fiber cell walls collapsed completely in further thermoforming, resulting in a closer alignment of the fiber filaments before the formation of high-strength physical and chemical bonding, expanded crystalline regions, and enhanced dimensional stability. The resulted biocomposites show higher ultimate tensile strength (5.31 times) and ultimate flexural strength (5.22 times) than that of Buxus sinica. In addition, the preparation method also endows the biocomposites with excellent hydrophobicity and high chemical stability towards concentrated strong acid and alkali. Overall, this study reveals a promising method to effectively transform the artificial shrub resources into high-quality biocomposites with versatile applications.
Extensive phytochemical study of the methanol extract of twigs and leaves of Buxus sinica resulted in the identification of forty-one Buxus alkaloids, including twenty undescribed ones, namely ...cyclobuxusinines A-I (1–7, 16 and 20), as well as secobuxusinines A-K (8–15 and 17–19). Their structures were delineated by detailed analysis using various spectroscopic techniques. cyclobuxusinines B (2) was the first Buxus alkaloid, whose CH3-18 was oxidized, implying the presence of special oxidative enzymes in this plant. Secobuxusinines C (10), D (11), and E (12), whose C-12 or C-19 have an OH group substitution, enriched the substituent pattern in Buxus alkaloid and suggested more structurally diverse alkaloids in the Buxus spp. In the assessment of their bioactivities, some of them exhibited significant cytotoxic effects on two human tumor ovarian cancer cell lines. Notably, compound 36 displayed more potent cytotoxic effect against ES2 and A2780 cell lines with the IC50 value of 1.33 μM and 0.48 μM, respectively.
Forty-one metabolites, including twenty undescribed Buxus alkaloids, were isolated from the aerial part of Buxus sinica. Some of them showed good cytotoxic activity. Display omitted
•1, Twenty undescribed and twenty-one known Buxus alkaloids were isolated from Buxus sinica.•2, Some Buxus alkaloids showed significant cytotoxicity against ES2 and A2780 cell lines.•3, Compound 36 displayed more potent cytotoxic with the IC50 value of 1.33 μM and 0.48 μM, respectively.
Amphetamine analogues are produced by plants in the genus Ephedra and by khat (Catha edulis), and include the widely used decongestants and appetite suppressants (1S,2S)-pseudoephedrine and ...(1R,2S)-ephedrine. The production of these metabolites, which derive from L-phenylalanine, involves a multi-step pathway partially mapped out at the biochemical level using knowledge of benzoic acid metabolism established in other plants, and direct evidence using khat and Ephedra species as model systems. Despite the commercial importance of amphetamine-type alkaloids, only a single step in their biosynthesis has been elucidated at the molecular level. We have employed Illumina next-generation sequencing technology, paired with Trinity and Velvet-Oases assembly platforms, to establish data-mining frameworks for Ephedra sinica and khat plants. Sequence libraries representing a combined 200,000 unigenes were subjected to an annotation pipeline involving direct searches against public databases. Annotations included the assignment of Gene Ontology (GO) terms used to allocate unigenes to functional categories. As part of our functional genomics program aimed at novel gene discovery, the databases were mined for enzyme candidates putatively involved in alkaloid biosynthesis. Queries used for mining included enzymes with established roles in benzoic acid metabolism, as well as enzymes catalyzing reactions similar to those predicted for amphetamine alkaloid metabolism. Gene candidates were evaluated based on phylogenetic relationships, FPKM-based expression data, and mechanistic considerations. Establishment of expansive sequence resources is a critical step toward pathway characterization, a goal with both academic and industrial implications.
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Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Protein enhancer of sevenless 2B, E(sev)2B, is a key adapter protein in the Ras/MAPK signaling pathway which has been reported to be involved in innate immunity. In this study, the gene that encodes ...AsE(sev)2B was isolated from A. sinica. It was found to contain a 636 bp open reading frame encoding 211 amino acids with a calculated molecular mass of 24.357 kDa and a predicted isoelectric point of 5.39. The predicted protein contains a N-terminal Src homology 3 domain (SH3), a central Src homology 2 domain (SH2), and a C-terminal Src homology 3 domain (SH3). Homology analysis revealed that AsE(sev)2B shares 49%–95% identity with E(sev)2B homologs from other species. In this study, the expression pattern and location of AsE(sev)2B during different stages of embryonic development and bacterial challenge were investigated by means of real-time qPCR, Western blotting and immunohistochemistry. Results showed that the highest expression level of AsE(sev)2B was at 0 h. After challenged by Gram-positive bacteria and Gram-negative bacteria, AsE(sev)2B was remarkably upregulated at 106 cellsL−1 bacterial concentrations. These results suggested that AsE(sev)2B plays a vital role during early embryonic development and in immune responses against bacterial challenge.
•Open reading frame of E(sev)2B was isolated and characterized from Artemia sinica.•AsE(sev)2B was expressed at all developmental stages using RT-qPCR and Western blotting.•The signals of AsE(sev)2B were mainly detected in the cephalothorax by immunohistochemistry.•AsE(sev)2B was remarkably upregulated under stimulated by Gram-positive and Gram-negative bacteria.
Phylogenetically diverse anammox bacteria have been detected in most of anoxic natural and engineered ecosystems and thus regarded as key players in the global nitrogen cycle. However, ecological ...niche differentiation of anammox bacteria remains unresolved despite its ecological and practical importance. In this study, the microbial competitions for a common substrate (nitrite) among three anammox species (i.e. “Candidatus Brocadia sinica”, “Candidatus Jettenia caeni” and “Candidatus Kuenenia stuttgartiensis”) were systematically investigated in nitrite-limited gel-immobilized column reactors (GICR) and membrane bioreactors (MBRs) under different nitrogen loading rates (NLRs). 16 S rRNA gene-based population dynamics revealed that “Ca. J. caeni” could proliferate only at low NLRs, whereas “Ca. B. sinica” outcompeted other two species at higher NLRs in both types of reactors. Furthermore, FISH analysis revealed that “Ca. J. caeni” was mainly present as spherical microclusters at the inner part (low NO2− environment), whereas “Ca. B. sinica” was present throughout the gel beads and granules. This spatial distribution supports the outcomes of the competition experiments. However, the successful competition of “Ca. J. caeni” at low NLR could not be explained with the Monod model probably due to inaccuracy of kinetic parameters such as half saturation constant (Ks) for nitrite and a difference in the maintenance rate (m). In addition, the growth of “Ca. K. stuttgartiensis” could not be observed in any experimental conditions, suggesting possible unknown factor(s) is missing. Taken together, NLR was one of factors determining ecological niche differentiation of “Ca. B. sinica” and “Ca. J. caeni”.
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•Competition for nitrite among three enriched anammox bacteria was systematically investigated.•“Ca. B. sinica” dominated at high NLRs, while “Ca. J. caeni” dominated at low NLRs.•Monod model could not predict the dominance of “Ca. J. caeni” at low NLRs.•NLR was a factor determining ecological niche differentiation of anammox bacteria.
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