Study on the technology of efficient extraction of eleutheroside E from Acanthopanax senticosus by green solvent DES.

OBJECTIVES: Acanthopanax senticosus (Rupr. & Maxim.) Harms is a medicinal and edible plant which is clinically used for the recovery and treatment of cardiovascular and central diseases. As a characteristic active pharmaceutical ingredient of Acanthopanax senticosus, eleutheroside E is the core of the therapeutic effect. Organic solvent extraction has low selectivity, low extraction rate, difficulty in separation and purification and safety risks. The purpose of this study was to extract the effective component of Acanthopanax senticosus with a new green solvent. METHODS: In this article, two kinds of deep eutectic solvents (DESs) (DES-1 and DES-2) were synthesised by heating and stirring methods. Eleutheroside E was extracted by ultrasonic extraction with two kinds of DES as extractants and quantitatively analysed by Orbitrap-tandem mass spectrometry (MS/MS). RESULTS: The main results showed that the initial polarity of the DES was similar to that of 60 to 80% ethanol and hydrogen bond donors were the main factors affecting the polarity of DES. In the test, the viscosity of DES was higher than that of ethanol, and even the addition of a small amount of water (10%) caused intermolecular hydrogen bond disruption and redistribution of the solvent, resulting in a significant decrease in solvent viscosity. The solvents in the test group were stable after standing at 5°C in the dark for 100 days. The extraction rate of eleutheroside E by DES solvent was 5-6 times higher than that by ethanol. DES-1 and DES-2 can efficiently extract eleutheroside E. CONCLUSION: This study developed a new method for the application of the green extraction of eleutheroside E with certain practical significance.

Pharmacokinetics and disposition of monoterpene glycosides derived from Paeonia lactiflora roots (Chishao) after intravenous dosing of antiseptic XueBiJing injection in human subjects and rats.

AIM: Monoterpene glycosides derived from Paeonia lactiflora roots (Chishao) are believed to be pharmacologically important for the antiseptic herbal injection XueBiJing. This study was designed to characterize the pharmacokinetics and disposition of monoterpene glycosides. METHODS: Systemic exposure to Chishao monoterpene glycosides was assessed in human subjects receiving an intravenous infusion and multiple infusions of XueBiJing injection, followed by assessment of the pharmacokinetics of the major circulating compounds. Supportive rat studies were also performed. Membrane permeability and plasma-protein binding were assessed in vitro. RESULTS: A total of 18 monoterpene glycosides were detected in XueBiJing injection (content levels, 0.001-2.47 mmol/L), and paeoniflorin accounted for 85.5% of the total dose of monoterpene glycosides detected. In human subjects, unchanged paeoniflorin exhibited considerable levels of systemic exposure with elimination half-lives of 1.2-1.3 h; no significant metabolite was detected. Oxypaeoniflorin and albiflorin exhibited low exposure levels, and the remaining minor monoterpene glycosides were negligible or undetected. Glomerular-filtration-based renal excretion was the major elimination pathway of paeoniflorin, which was poorly bound to plasma protein. In rats, the systemic exposure level of paeoniflorin increased proportionally as the dose was increased. Rat lung, heart, and liver exposure levels of paeoniflorin were lower than the plasma level, with the exception of the kidney level, which was 4.3-fold greater than the plasma level; brain penetration was limited by the poor membrane permeability. CONCLUSION: Due to its significant systemic exposure and appropriate pharmacokinetic profile, as well as previously reported antiseptic properties, paeoniflorin is a promising XueBiJing constituent of therapeutic importance.

[Mechanism of biological actions of quercetin based on biomolecular network].

The mechanism of biological actions of quercetin was studied by using metabolomic method and biomolecular network. HPLC-MS was used to analyze the serum metabolome in rats of blank group and quercetin administration group rats, and MS data were processed by MATLAB software. With multivariate statistical analysis of serum metabolite profiles, a clear separation among blank group and quercetin administration group was achieved, potential biomarkers were selected according to the parameters of variable importance in the projection (VIP) and identified according to MS information and database retrieval. Four compounds, related enzymes, action targets and metabolic pathways had been confirmed, namely retinoic acid and RARbeta, arachidonate and COX-2, 3, 5-diodotyrosine and TPO, uridine diphosphate glucose and PDEs. The mechanism of quercetin enhancing ability of retinoic acid on the induction of RARbeta, activating TPO, using as COX-2 and PDEs inhibitor was approved by biomolecular network and related literatures. In this study, a mechanism of multiple biological actions of quercetin was evaluated at the level of the biomolecular network, metabolomics and biomolecular network can be used to investigate the biological effects mechanism of quercetin, which provided a new method to further revealing mechanism of drug action.

The Stenopsyche simplex Species Group from China with descriptions of three new species (Trichoptera: Stenopsychidae).

The Chinese Stenopsyche simplex Group is revised for the first time since Weaver's (1987) revision. The group includes 14 described species and 3 new species. Two diagnostic subgroups were recognized by Schmid (1959). The first subgroup of Chinese species includes S. anaximander Malicky 2011, S. brevata Tian & Zheng 1989, S. chinensis Hwang 1957, S. dentata Navás 1930, S. splendida Martynov 1935, S. dubia Schmid 1965, S. rotundata Schmid 1965, S. simplex Schmid 1959, and S. tienmushanensis Hwang 1957; of these, S. dentata is assigned to this first subgroup for the first time. The second diagnostic subgroup includes S. denticulata Ulmer 1926, S. longispina Ulmer 1926, S. stoetzneri Döhler 1929, S. uniformis Schmid 1965 and S. formosana Kobayashi, 1987. The species newly described here include Stenopsyche ningshanensis sp. nov., from Shaan-xi Province, belonging to the first subgroup, and Stenopsyche acanthoclada sp. nov. and Stenopsyche jinxiuensis sp. nov., both from Guang-xi Province and belonging to the second subgroup. Distribution maps of the two diagnostic subgroups of Chinese species and illustrations of male genitalia of the three new species are provided along with collection data for additional specimens for the previously described species.

Seven new species and four new records of Psychomyiidae (Insecta, Trichoptera) from China.

Seven new species of the family Psychomyiidae Walker, 1852 are described and illustrated from China; they are Psychomyiashunisp. nov., Ps.mangshanensissp. nov., Ps.capricornissp. nov., Lypesagittalissp. nov., Paduniellafasciariasp. nov., Pa.sanyaensissp. nov., and Tinodesaviformissp. nov. The genus Lype is reported for the first time from mainland China. In addition, four psychomyiids are found to be new to the Chinese caddis fauna: Psychomyiaindra Malicky & Chantaramongkol, 1993; Paduniellaandamanensis Malicky, 1979; Pa.dendrobia Malicky & Chantaramongkol, 1993; and Tinodesgapbona Johanson & Oláh, 2008. Moreover, Psychomyiapolyacantha Li, Qiu & Morse, 2021 is reviewed and synonymized with Psychomyiaimamiah Malicky, 2020.

Five new species of Tinodes (Trichoptera: Psychomyiidae) from China.

Five new species of genus Tinodes Curtis 1834 from China are described, illustrated, and diagnosed based on distinctive characters of male genitalia, including Tinodes falcata n. sp., T. xuzhouorum n. sp., T. shunhuangshanensis n. sp., T. yangae n. sp., and T. lini n. sp. Tinodes falcata n. sp. differs from other members of Tinodes in having an unpaired falcate and strongly sclerotized phallic sheath process. Tinodes xuzhouorum n. sp. differs from T. charmi Malicky 2009 by the relative lengths of the superior appendages and the phallus, the position of the ejaculatory duct, and the angle formed by tergum IX and the superior appendages in lateral view. Tinodes shunhuangshanensis n. sp. can be distinguished from T. sartael Malicky 2017 by the phallic sheath process splitting into paired dorsal branches and an apically incised ventral branch. Tinodes yangae n. sp. differs from T. eris Malicky & Sangpradub 2001 by the dorsal phallic sheath process having extra paired vertical branches. Tinodes lini n. sp. differs from T. dactringa Johanson & Oláh 2008 by the phallic guide with a pair of closely attached dorsal branches.

New species of the genus Molanna Curtis, 1834 (Trichoptera, Molannidae) in China inferred from morphology and DNA barcodes.

The male adult of Molannatruncata Ge, Peng & Sun sp. nov. is described and illustrated based on material collected in Si-chuan, China. It could be diagnosed by the subtriangular superior appendages when viewed dorsally, and by the mesal appendages each having a slender thorn and inferior appendages with a tiny inner process. Based on morphology of genitalia, we provide a dichotomous key to adult males of Molanna from the Oriental region. The DNA barcodes (partial mtCOI sequences) of M.truncata sp. nov. are generated and compared with existing sequences of Molanna species from Oriental and Palearctic regions. The mean intraspecific divergence of Molanna was 1.58% with a maximum of 8.50% in M.moesta. The Automatic Barcode Gap Discovery (ABGD) analysis of Molanna inferred 9 OTUs and thresholds of interspecific divergence of 10%. Divergence of M.truncata sp. nov. haplotypes from all other Molanna haplotypes ranged from 10.1% to 18%. We discuss distribution and potential groups of species within the Oriental Molanna species based on morphology.

Description of adult female, larva, and pupa of Glossosoma (Lipoglossa) kamael Malicky 2012 (Trichoptera: Glossosomatidae) associated by COI sequences from China.

The adult female, larva, and pupa of Glossosoma (Lipoglossa) kamael Malicky 2012 from Qinghai Province, China, are described and illustrated. Molecular associations are based on COI sequences. The immature stages of the subgenus Lipoglossa are described for the first time. Biological and habitat information of G. kamael are presented.

Fasudil, a Rho kinase inhibitor, drives mobilization of adult neural stem cells after hypoxia/reoxygenation injury in mice.

Rho kinase (ROCK) is important in fundamental processes of cell proliferation and survival. Blockade of ROCK promotes stem cell survival in vitro and axonal regeneration in vivo, exhibiting therapeutic potential such as spinal cord injuries and stroke. Here, we used the model of hypoxia/reoxygenation (H/R) injury to explore the possibility whether Fasudil, a ROCK inhibitor in clinical application for subarachnoid hemorrhage and stroke, mobilizes adult neural stem cells in vivo. Most interestingly, Fasudil triggers neurogenesis especially in the subventricular zone after H/R. The increase of Brdu+ cholinergic neurons was observed in striatum and forebrain cortex of Fasudil-treated mice after 30 days. Further observation demonstrates that both levels of granulocyte colony-stimulating factor (G-CSF) and astrocytes expressing G-CSF were elevated in mice treated with Fasudil, as compared to mice injected with saline. In vitro H/R model of cultured astrocytes, Fasudil promoted astrocytes to produce G-CSF in a dose-dependent manner. In addition, antibody neutralization and receptor blocking of the G-CSF pathway clearly demonstrate that Fasudil-induced neurogenesis was mediated partially through astrocyte-derived G-CSF. Our results indicate that Fasudil might represent a promising therapeutic perspective by mobilizating endogenous adult neural stem cells in the CNS.

Fasudil protects hippocampal neurons against hypoxia-reoxygenation injury by suppressing microglial inflammatory responses in mice.

Rho kinase (ROCK) may play an important role in regulating biological events of cells, including proliferation, differentiation and survival/death. Blockade of ROCK promotes axonal regeneration and neuron survival in vivo and in vitro, thereby exhibiting potential clinical applications in spinal cord damage and stroke. Our previous studies have demonstrated that Fasudil, a selective ROCK inhibitor, induced neuroprotection in vitro. Here we used an in vivo model of hypoxia/reoxygenation (H/R) injury to examine the neuroprotective effect of Fasudil, and explore its possible mechanism(s) in vivo. H/R resulted in the loss of hippocampal neurons, accompanied by increased apoptosis of neurons in hippocampus. The expression of ROCK II and activity of ROCK in the brain were increased after H/R, and located only in microglia, but not in astrocytes and neurons. The administration of Fasudil inhibited the activity of ROCK in brain tissue and cultured microglia, and protected hippocampal neurons against H/R injury. Further immunohistochemical analysis and cytokine determination revealed that Fasudil inhibited inducible nitric oxide synthase immunoreactivity in microglia and pro-inflammatory factors in brain tissue after H/R, which is consistent with the observation wherein Fasudil reduced the pro-inflammatory factors nitric oxide, IL-1ß, IL-6 and TNF-, and increased anti-inflammatory factor IL-10 in cultured microglia under normoxic or hypoxic conditions. Our results indicate that inhibition of ROCK by Fasudil may represent a useful therapeutic perspective by inhibiting microglial inflammatory responses in the CNS.

Therapeutic potential of experimental autoimmune encephalomyelitis by Fasudil, a Rho kinase inhibitor.

The migration of aberrant inflammatory cells into the central nervous system plays an important role in the pathogenesis of demyelinating diseases potentially through the Rho/Rho-kinase (Rock) pathway, but direct evidence from human and animal models remains inadequate. Here we further confirm that Fasudil, a selective Rock inhibitor, has therapeutic potential in a mouse model of myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis (EAE). The results show that Fasudil decreased the development of EAE in C57BL/6 mice. Immunohistochemistry disclosed that expression of Rock-II in the perivascular spaces and vascular endothelial cells of spleens, spinal cords, and brains was elevated in EAE and was inhibited in the Fasudil-treated group. T-cell proliferation specific to MOG(35-55) was markedly reduced, together with a significant down-regulation of interleukin (IL)-17, IL-6, and MCP-1. In contrast, secretion of IL-4 was increased, and IL-10 was slightly elevated. There were no differences in the percentages of CD4(+)CD25(+), CD8(+)CD28(-), and CD8(+)CD122(+) in mononuclear cells. Histological staining disclosed a marked decrease of inflammatory cells in spinal cord and brain of Fasudil-treated mice. These results, together with previous studies showing the inhibitory effect of Fasudil on T-cell migration, might expand its clinical application as a new therapy for multiple sclerosis by decreasing cell migration and regulating immune balance.

Four new species of Psychomyia (Trichoptera: Psychomyiidae) from China, with re-descriptions of two species.

Four new species of the genus Psychomyia from China are described, illustrated and diagnosed: P. complexa n. sp. (Zhe-jiang), P. conoidea n. sp. (Zhe-jiang), P. obtorta n. sp. (Jiang-xi), and P. tripetala n. sp. (Jiang-xi). Two known species, P. martynovi Hwang 1957 and P. extensa Li, Sun Yang 1999 are re-described and re-illustrated, with new distributional data.

Descriptions of larvae of three Philopotamidae species from China (Insecta, Trichoptera).

Adults and larvae of the family Philopotamidae from Zhejiang Province, China, were examined and mtCOI gene sequences were extracted and analyzed, males and larvae of 3 species were successfully associated. The larvae of Chimarra sadayu Malicky 1993, Dolophilodes bellatula Sun Malicky 2002, Wormaldia unispina Sun 1998 are described in detail and their diagnostic photographs and illustrations are presented. Diagnostic characters for genera and species are discussed.

A new species of Micardia Butler, 1878 (Lepidoptera, Noctuidae, Eustrotiinae) from China.

The genus Micardia Butler, 1878 was described based on M. argentata (the type species; Fig. 1) and M. pulchra, both from Japan; in the same paper, Buter transferred a species described from India, Leucania pulcherrima Moore, 1867 to Micardia. Hampson (1990) considered Micardia a synonym of Eustrotia. However, its generic status was revised by Warren (1913) and retained by subsequent authors (Ueda 1984). There are at present 13 species recognized within Micardia (Table 1), of which, M. quadrilinea Scriba 1921 was described from Japan on the basis of female specimens and without high-quality illustrations, and subsequently treated as of uncertain status by Chen Xue (2012). The remaining species are as follows: M. simplicissima Berio 1973 was described from Myanmar based on 2 females. M. munda Leech, 1900, was described from Western China (Sichuan, according to Chen Xue 2012). Chen (1999) extended the range of M. pulcherrima to include China, and Chen Xue (2012) subsequently reviewed Micardia from China, describing three new species, M. pallens, M. distincta and M. minuta. Four species have been described from Madagascar, M. argentoidea and M. terracottoides by Berio (1954), and M. ikoly and M. itremo by Viette (1982). Because they are geographically separated so widely from their congeners in Asia, Chen Xue (2012) considered their generic placement in need of confirmation. Moreover, the distribution of M. pulchra Butler was extended to Russia (Kurile Island) by synonymizing it with M. pulchrargentea Bryk, 1942 (Kononenko 2005). In this paper we report a new species, Micardia yixingensis, sp. nov. from Jiangsu, China. The contribution brings the total number of the species in the genus to 14.

A new species of Chimarra from China (Trichoptera, Philopotamidae) with description of its larva.

The adult male and larva of a new species in the genus Chimarra Stephens, Chimarra paramonorum n. sp., are described, diagnosed, and illustrated based on the specimens collected in Zhejiang Province, China. The larva and adult male were associated by mtCOI gene sequences. The male can be separated from C. monorum and C. thienemanni mainly by small preanal appendages and by wrinkled, hairless endotheca. The larva of the new species can be separated from other congeners by a combination of characters including the notch on the anterior margin of the frontoclypeus, the teeth of the mandibles, and the length of the seta-bearing process of each forecoxa. The larval description is the first for Chimarra in China.

Descriptions of larvae of three species of Hydropsyche (Trichoptera, Hydropsychidae) from China.

Twenty-one individuals of Hydropsyche from An-hui and Zhe-jiang Provinces, China, were examined and their DNA barcode sequences were generated and analyzed. The larvae and males of H. arion, H. columnata, H. simulata, and H. trifora were associated by mtCOI gene sequences. The larvae of H. arion, H. columnata, and H. trifora are described in detail and their diagnostic photographs are provided. Character photographs of the H. simulata larva are also presented. A key to 5th instar larvae of 15 species of Chinese Hydropsyche is provided.

Fragmentation characteristics of hydroxycinnamic acids in ESI-MSn by density functional theory.

This work aims to analyze the electrospray ionization multistage mass spectrometry (ESI-MSn ) fragmentation characteristics of hydroxycinnamic acids (HCAs) in negative ion mode. The geometric parameters, energies, natural bond orbitals and frontier orbitals of fragments were calculated by density functional theory (DFT) to investigate mass spectral fragmentation mechanisms. The results showed that proton transfer always occurred during fragmentation of HCAs; their quasi-molecular ions ([M - H]- ) existed in more than one form and were mainly with the lowest energy. The fragmentation characteristics included the followings: (1) according to the different substitution position of phenolic hydroxyl group, the ring contraction reaction by CO elimination from benzene was in an increasingly difficult order: m-phenolic hydroxyl > p-phenolic hydroxyl > o-phenolic hydroxyl; and (2) ortho effect always occurred in o-dihydroxycinnamic acids (o-diHCAs), i.e. one phenolic hydroxyl group offered H+ , which combined with the other one to lose H2 O. In addition, there was a nucleophilic reaction during ring contraction in diHCAs that oxygen atom attacked the carbon atom binding with the other phenolic hydroxyl to lose CO2 . The fragmentation characteristics and mechanism of HCAs could be used for analysis and identification of such compounds quickly and effectively, and as reference for structural analogues by ESI-MS. Copyright © 2017 John Wiley & Sons, Ltd.

Descriptions of pupae of two Hydropsyche species from China (Insecta, Trichoptera, Hydropsychidae).

Pupae of two Hydropsyche species collected from Zhe-jiang Province, China, were examined and their barcode sequences were generated and analyzed. The pupae of H. simulata and H. homunculus were associated with their adult males by mtCOI gene sequences, described, illustrated, and diagnosed.

Notes on the Rhyacophila scissa species group with description of two new taxa from China (Trichoptera, Rhyacophilidae).

One new species, R. meniscoides n. sp., and 1 new subspecies, R. morsei concaviuscula n. sp., are described, diagnosed and illustrated. Two diagnostic species complexes within the R. scissa Species Group, the R. scissa Diagnostic Species Complex and the R. schismatica Diagnostic Species Complex, are recognized and delimited. The 15 previously described species and 1 new species and 1 new subspecies described in this article are assigned to the complexes accordingly. Distribution maps of all 17 members in the Species Group are presented.