Determination of flurbiprofen in rat plasma using ultra-high performance liquid chromatography-tandem mass spectrometry and its application in a pharmacokinetic study.

A rapid and sensitive ultra-high performance liquid chromatography-tandem mass spectrometry method was developed to determine flurbiprofen in rat plasma. A triple quadrupole tandem mass spectrometer equipped with an electrospray ionization (ESI) source was used in negative ion mode. Acetonitrile precipitation was selected to prepare samples. Flurbiprofen and internal standard flurbiprofen-d5 were analyzed on an Acquity UPLC BEH C18 column with the mobile phase consisting of acetonitrile and water, and a gradient procedure was used for separation. The retention time of flurbiprofen was 0.67 min, and the whole running time was only 1.2 min. The detection was performed on a triple quadrupole tandem mass spectrometer using multiple reaction monitoring mode via an ESI source with optimized mass spectrometry parameters. The calibration curve was linear in the range of 25.0-1.00 × 104 ng/mL (r ≥ 0.99). The within-run and between-run relative standard deviations were not more than 13.9%. The within-run and between-run relative errors were from -9.0% to 3.4%. There was no significant matrix effect, and recovery was high. This method was fully validated, including whole blood stability in rat plasma, and successfully applied to the pharmacokinetic study in which 100% incurred sample reanalysis met the criteria.

New Cretaceous Lacewings in a Transitional Lineage of Myrmeleontoidea and Their Phylogenetic Implications.

The extinct neuropteran families Cratosmylidae and Babinskaiidae hitherto only known from the Cretaceous represent the transitional lineage between Nymphidae and advanced myrmeleontoids (e.g., Nemopteridae and Myrmeleontidae) in the superfamily Myrmeleontoidea. Here, we describe two new species, which respectively belong to Cratosmylidae and Babinskaiidae, namely, Araripenymphes burmanus sp. nov. and Paradoxoleon chenruii gen. et sp. nov., from the mid-Cretaceous Kachin amber of Myanmar. Cratosmylidae, which was previously only recorded from the Lower Cretaceous of Brazil (Crato Formation), is first reported from the mid-Cretaceous Kachin amber of Myanmar, and the co-occurrence of Araripenymphes Menon, Martins-Neto and Martill, 2005 across South America and Asia further documents the Gondwanan origin of the northern Myanmar amber lacewing paleofauna. The first finding of a deeply bifurcated forewing MP with two free branches in Babinskaiidae (viz., Paradoxoleon chenruii gen. et sp. nov.) highlights the morphological diversity of this extinct family. The phylogenetic positions of Araripenymphes burmanus sp. nov. and Paradoxoleon chenruii gen. et sp. nov. were recovered on the basis of a morphology-based phylogenetic analysis, and the monophyly of Cratosmylidae + Babinskaiidae was corroborated. Given the paraphyly of Cratosmylidae, its familial status is discussed.

The first mitochondrial genome of spongillafly from Asia (Neuroptera: Sisyridae: Sisyra aurorae Navás, 1933) and phylogenetic implications of Osmyloidea.

The spongillafly species Sisyra aurorae Navás, 1933 (Neuroptera: Sisyridae) is an endemic species in China and is first recorded from Shanghai. The mitogenome of this species is sequenced, representing the first mitogenome of Sisyridae from Asia. The nearly complete mitogenome is 15,634 bp, which contains 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs), and a control region. The gene order and arrangement are similar to other lacewing mitogenomes. Both Bayesian and maximum likelihood analyses based on 13 PCGs recovered the interfamilial phylogeny within Osmyloidea as Sisyridae + (Nevrorthidae + Osmylidae).

New Cretaceous antlion-like lacewings promote a phylogenetic reappraisal of the extinct myrmeleontoid family Babinskaiidae.

Babinskaiidae is an extinct family of the lacewing superfamily Myrmeleontoidea, currently only recorded from the Cretaceous. The phylogenetic position of this family is elusive, with inconsistent inferences in previous studies. Here we report on three new genera and species of Babinskaiidae from the mid-Cretaceous Kachin amber of Myanmar, namely Calobabinskaia xiai gen. et sp. nov., Stenobabinskaia punctata gen. et sp. nov., and Xiaobabinskaia lepidotricha gen. et sp. nov. These new babinskaiids are featured by having specialized characters, such as the rich number of presectoral crossveins and the presence of scaly setae on forewing costal vein, which have not yet been found in this family. The exquisite preservation of the Kachin amber babinskaiids facilitate a reappraisal of the phylogenetic placement of this family based on adult morphological characters. Our result from the phylogenetic inference combining the data from fossil and extant myrmeleontoids recovered a monophyletic clade composed of Babinskaiidae and another extinct family Cratosmylidae, and further assigned this clade to be sister group to a clade including Nemopteridae, Palaeoleontidae, and Myrmeleontidae. Babinskaiidae appears to be a transitional lineage between Nymphidae and advanced myrmeleontoids, with ancient morphological diversification.

Cretaceous diversity and disparity in a lacewing lineage of predators (Neuroptera: Mantispidae).

Mantidflies (Mantispidae) are an unusual and charismatic group of predatory lacewings (Neuroptera), whereby the adults represent a remarkable case of morphological and functional convergence with praying mantises (Mantodea). The evolutionary history of mantidflies remains largely unknown due to a scarcity of fossils. Here, we report the discovery of a highly diverse palaeofauna of mantidflies from the mid-Cretaceous (lowermost Cenomanian) of Myanmar. The raptorial forelegs of these mantidflies possess highly divergent morphological modifications, some of which are unknown among modern mantidflies, e.g. the presence of forked basal profemoral spines or even the complete loss of foreleg spine-like structures. A phylogenetic analysis of Mantispidae reveals a pattern of raptorial foreleg evolution across the family. The high species diversity and disparate foreleg characters might have been driven by diverse niches of predator-prey interplay in the complex tropical forest ecosystem of the mid-Cretaceous.

A simple and low-energy method to prepare loratadine nanosuspensions for oral bioavailability improvement: preparation, characterization, and in vivo evaluation.

The effervescent method, as a simple and effective technology to prepare nanosuspensions, has gained great attention. In this present research, loratadine (LTD) nanosuspensions were successfully prepared by the effervescent method using Soluplus as stabilizer to improve the bioavailability of LTD in vivo. The mean particle size was about 100 nm. And the LTD nanosuspensions were lyophilized for further study. The freeze-dried powders could be dissolved quickly, and the mean particle size remained almost unchanged after powders were re-dissolved. By transmission electron microscope (TEM), differential scanning calorimetry (DSC), scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy, and X-ray diffraction (X-RD), the characterizations of LTD nanosuspensions and freeze-dried powders were studied. Commercial tablets were used as the reference to investigate the dissolution behaviors in different release media and of bioavailability in vivo of LTD freeze-dried powders. The cumulative dissolution of the LTD freeze-dried powders was superior in different release media compared with commercial tables. In addition, for the evaluation of the bioavailability of LTD nanosuspensions, the LTD concentration in rat plasma was determined using LC-MS/MS method. The results showed that the AUC0-24 and Cmax of LTD freeze-dried powders were about 2.14- and 2.01-fold higher than those of commercial tablets. In short, the effervescent method has been successfully applied to the preparation of LTD nanosuspensions to improve the bioavailability of LTD in vivo with the advantage of low energy consumption. This simple technology also provides an idea for the preparation of the other nanosuspensions.

Xanthoceraside Could Ameliorate Alzheimer's Disease Symptoms of Rats by Affecting the Gut Microbiota Composition and Modulating the Endogenous Metabolite Levels.

Xanthoceraside (XAN) is a natural-derived compound with anti-Alzheimer activity from the husks of Xanthoceras sorbifolia. Although its therapeutic effect had been confirmed in previous studies, the mechanism was still unclear due to its poor solubility and low permeability. In this study, the pharmacological effect of XAN on Alzheimer's disease (AD) was confirmed by behavior experiments and H&E staining observation. Fecal microbiota transplantation (FMT) experiment also replicated the therapeutic effects, which indicates the potential targets of XAN on gut microbiota. The sequencing of 16S rRNA genes in fecal samples demonstrated that XAN reversed gut microbiota dysbiosis in AD animals. XAN could change the relative abundances of several phyla and genus of bacterial, particularly the ratio of Firmicutes/Bacteroidetes. Among them, Clostridium IV, Desulfovibrio, Corynebacterium, and Enterorhabdus had been reported to be involved in the pathologic developments of AD and other central nervous system disease. In metabolomics study, a series of host endogenous metabolites were detected, including amino acids, lysophosphatidylcholine, dihydrosphingosine, phytosphingosine, inosine, and hypoxanthine, which were all closely associated with the development of AD. Combined with the Spearman's correlation analysis, it was confirmed that the increases of five bacterial strains and decreases of six bacterial strains were closely correlated with the increases of nine host metabolites and the decreases of another five host metabolites. Therefore, XAN can modulate the structure of gut microbiota in AD rats; the changes of gut microbiota were significantly correlated with endogenous metabolites, and symptom of AD was ultimately alleviated. Our findings suggest that XAN may be a potential therapeutic drug for AD, and the gut microbiota may be potential targeting territory of XAN via microbiome-gut-brain pathway.

High niche diversity in Mesozoic pollinating lacewings.

Niche diversity of pollinating insects plays a vital role in maintaining extant terrestrial ecosystems. A key dimension of pollination niches refers to the insect proboscis length that commonly matches the floral tube length. Here we describe new kalligrammatid lacewings (an iconic Mesozoic pollinating insect lineage) from late Cretaceous Burmese amber and Mesozoic sediments in China. Kalligrammatids display complex configurations of elongate mouthpart elements consisting of well-developed maxillae, labium and their palps. The mouthpart lengths vary among species, from 0.6 to 18.0 mm, suggesting corresponding variability in the floral tube lengths of Mesozoic plants. With the diversification of pollinating habits, the kalligrammatids presented highly divergent traits related to chemical communication and defence mechanisms. Together with other Mesozoic long-proboscid insects, these fossils not only reveal the high niche diversity of Mesozoic pollinating insects but also highlight the diversity of Mesozoic pollinator-dependent plants prior to the rise of angiosperms.

Liverwort Mimesis in a Cretaceous Lacewing Larva.

Camouflage and mimicry are staples among predator-prey interactions, and evolutionary novelties in behavior, anatomy, and physiology that permit such mimesis are rife throughout the biological world [1, 2]. These specializations allow for prey to better evade capture or permit predators to more easily approach their prey, or in some cases, the mimesis can serve both purposes. Despite the importance of mimesis and camouflage in predator-avoidance or hunting strategies, the long-term history of these traits is often obscured by an insufficient fossil record. Here, we report the discovery of Upper Cretaceous (approximately 100 million years old) green lacewing larvae (Chrysopoidea), preserved in amber from northern Myanmar, anatomically modified to mimic coeval liverworts. Chrysopidae are a diverse lineage of lacewings whose larvae usually camouflage themselves with a uniquely constructed packet of exogenous debris, conveying greater stealth upon them as they hunt prey such as aphids as well as evade their own predators [3, 4]. However, no lacewing larvae today mimic their surroundings. While the anatomy of Phyllochrysa huangi gen. et sp. nov. allowed it to avoid detection, the lack of setae or other anatomical elements for entangling debris as camouflage means its sole defense was its mimicry, and it could have been a stealthy hunter like living and other fossil Chrysopoidea or been an ambush predator aided by its disguise. The present fossils demonstrate a hitherto unknown life-history strategy among these "wolf in sheep's clothing" predators, one that apparently evolved from a camouflaging ancestor but did not persist within the lineage.

Taxonomic notes on Cretarophalis patrickmuelleri Wichard, 2017 (Insecta: Neuroptera: Nevrorthidae) from the mid-Cretaceous of Myanmar, and its phylogenetic significance.

Nevrorthidae is a family of Neuroptera with low species diversity, disjunct geographic distributions, and a controversial phylogenetic status. Previous fossil records of the family are from the Eocene except for the recently described species Cretarophalis patrickmuelleri Wichard, 2017 from mid-Cretaceous Burmese amber, following earlier records of an undescribed species. However, such a significant finding of this family from Mesozoic was originally presented only with a preliminary description. Here we re-describe the species based on exquisitely preserved materials from mid-Cretaceous Burmese amber that provides for a much more detailed description, in particular of the hindwing, female genitalia, and larval head. Furthermore, distinctive morphological characters of C. patrickmuelleri with significant phylogenetic relevance are discussed. The general morphology, particularly that of the larva, has changed little between the Mesozoic and Cenozoic, and it reflects an evolutionary stasis that might correspond to their aquatic larval life-style. The present finding also supports the relictual nature of the modern nevrorthids.

The first green lacewing (Insecta: Neuroptera: Chrysopidae) from the mid-Cretaceous amber of Myanmar.

The lacewing family Chrysopidae is here reported from the mid-Cretaceous amber of Myanmar for the first time. A new genus and species, Parabaisochrysa xingkei gen. et sp. nov., is described. The new genus belongs to the extinct subfamily Limaiinae and is characterized by the presence of three gradate series of crossveins in both fore- and hind wing, and by the MP1 coalescent with RP+MA in the hind wing.

Fine Mapping of CsVYL, Conferring Virescent Leaf Through the Regulation of Chloroplast Development in Cucumber.

Leaf color mutants in higher plants are ideal materials for investigating the structure and function of photosynthetic system. In this study, we identified a cucumber vyl (virescent-yellow leaf) mutant in the mutant library, which exhibited reduced pigment contents and delayed chloroplast development process. F2 and BC1 populations were constructed from the cross between vyl mutant and cucumber inbred line 'Hazerd' to identify that the vyl trait is controlled by a simply recessive gene designated as CsVYL. The CsVYL gene was mapped to a 3.8 cM interval on chromosome 4 using these 80 F2 individuals and BSA (bulked segregation analysis) approach. Fine genetic map was conducted with 1542 F2 plants and narrowed down the vyl locus to an 86.3 kb genomic region, which contains a total of 11 genes. Sequence alignment between the wild type (WT) and vyl only identified one single nucleotide mutation (C→T) in the first exon of gene Csa4G637110, which encodes a DnaJ-like zinc finger protein. Gene Expression analysis confirmed the differences in transcription level of Csa4G637110 between wild type and mutant plants. Map-based cloning of the CsVYL gene could accelerate the study of chloroplast development and chlorophyll synthesis of cucumber.

Taxonomic notes on Babinskaiidae from the Cretaceous Burmese amber, with the description of a new species (Insecta, Neuroptera).

Babinskaiidae is an extinct lacewing family of the superfamily Myrmeleontoidea. Hitherto, nine species of seven genera are described from the Lower and mid-Cretaceous. Here a new species of Babinskaiidae is described from Cretaceous Burmese amber, namely Parababinskaia makarkinisp. n. The new species possesses an A2 vein in the hind wing, suggesting that the loss of this vein might not be an autapomorphy of Babinskaiidae. The female of Electrobabinskaia burmana Lu, Zhang & Liu, 2017 is also described for the first time based on two specimens with their abdomens perfectly preserved, exhibiting a specialised sternum VI with paired elongate projections. A brief discussion of female genital characters is provided, which may increase our understanding of the morphology and phylogenetic position of Babinskaiidae.

First description of female of Haplosialodes liui Huang et al., 2016 (Megaloptera: Sialidae) from Cretaceous Burmese amber.

A female of the alderfly species, Haplosialodes liui Huang et al., 2016, from the mid-Cretaceous Burmese amber is described for the first time. Systematic position of the genus Haplosialodes Huang et al., 2016 is briefly discussed.

First description of female of Haplosialodes liui Huang et al., 2016 (Megaloptera: Sialidae) from Cretaceous Burmese amber.

A female of the alderfly species, Haplosialodes liui Huang et al., 2016, from the mid-Cretaceous Burmese amber is described for the first time. Systematic position of the genus Haplosialodes Huang et al., 2016 is briefly discussed.

New genera and species of the minute snakeflies (Raphidioptera: Mesoraphidiidae: Nanoraphidiini) from the mid Cretaceous of Myanmar.

The Mesozoic amber snakeflies are important for understanding the early evolution and phylogeny of Raphidioptera. Here we describe three new genera and four new species of the extinct family Mesoraphidiidae from the mid Cretaceous amber of northern Myanmar, including Dolichoraphidia aspoecki gen. et sp. nov., Dolichoraphidia engeli gen. et sp. nov., Rhynchoraphidia burmana gen. et sp. nov., and Burmoraphidia reni gen. et sp. nov. All these species, together with the previously described species (i.e., Nanoraphidia electroburmica Engel, 2002) from Burmese amber, belong to the mesoraphidiid tribe Nanoraphidiini based on the minute body-size and the swollen tibiae. A key to all species of Nanoraphidiini is provided. A brief discussion on the genital morphology of Mesoraphidiidae is also given.

Serum metabonomics study of anti-depressive effect of Xiao-Chai-Hu-Tang on rat model of chronic unpredictable mild stress.

Xiao-Chai-Hu-Tang (XCHT) has been proven to be effective for the clinical treatment of depression. However, the mechanisms of definite antidepressant-like effects and detailed metabolic biomarkers were still unclear in this prior study. Here, we have investigated the metabolic profiles and potential biomarkers in a chronic unpredictable mild stress model after treatment with XCHT. Metabonomics based on ultra-high performance liquid chromatography coupled with mass spectrometry was used to profile the metabolic fingerprints of serum obtained from a rat model with chronic unpredictable mild stress with and without XCHT treatment. The model rats showed a significant decrease in sucrose preference and food consumption, and these depression-like symptoms were significantly improved by XCHT. Through principal component analysis (PCA), nine potential biomarkers of tryptophan, uric acid, phenylalanine, cholic acid and lysophosphatidylcholine (C18:0 LPC, C16:0 LPC, C16:1 LPC, C18:1 LPC, C20:4 LPC) were characterized as potential biomarkers involved the pathogenesis of depression. The therapeutic effect of XCHT on depression may involve in amino acid metabolism, lipid metabolism, oxidative stress and inflammation response. The present investigation highlights that metabonomics is a valuable tool for studying the essence of depression as well as evaluating the efficacy of the corresponding drug treatment.

New long-proboscid lacewings of the mid-Cretaceous provide insights into ancient plant-pollinator interactions.

Many insects with long-proboscid mouthparts are among the pollinators of seed plants. Several cases of the long-proboscid pollination mode are known between fossil insects (e.g., true flies, scorpionflies, and lacewings) and various extinct gymnosperm lineages, beginning in the Early Permian and increasing during the Middle Jurassic to Early Cretaceous. However, details on the morphology of lacewing proboscides and the relevant pollination habit are largely lacking. Here we report on three lacewing species that belong to two new genera and a described genus from mid-Cretaceous (Albian-Cenomanian) amber of Myanmar. All these species possess relatively long proboscides, which are considered to be modified from maxillary and labial elements, probably functioning as a temporary siphon for feeding on nectar. Remarkably, these proboscides range from 0.4-1.0 mm in length and are attributed to the most diminutive ones among the contemporary long-proboscid insect pollinators. Further, they clearly differ from other long-proboscid lacewings which have a much longer siphon. The phylogenetic analysis indicates that these Burmese long-proboscid lacewings belong to the superfamily Psychopsoidea but cannot be placed into any known family. The present findings represent the first description of the mouthparts of long-proboscid lacewings preserved in amber and highlight the evolutionary diversification of the ancient plant-pollinator interactions.

A novel LC-MS/MS assay for the simultaneous determination of melatonin and its two major metabolites, 6-hydroxymelatonin and 6-sulfatoxymelatonin in dog plasma: Application to a pharmacokinetic study.

A convenient and specific liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous quantification of melatonin (MEL) and its major metabolites, 6-hydroxymelatonin (6-O-MEL) and 6-sulfatoxymelationin (S-O-MEL) in dog plasma. After plasma samples were deproteinized with acetonitrile, the post-treatment samples were analyzed on a Phenomenex Kinetex C18 column (50×2.1 mm, 1.7 µm) interfaced with a triple quadrupole tandem mass spectrometer. Electrospray ionization mode (ESI) and multiple reaction monitoring were used to assay MEL and its metabolites. Acetonitrile and 5 mM ammonium acetate were used as the mobile phase with a gradient elution at a flow rate of 0.2 mL/min. The analytical run time of 6.5 min was divided into two periods according to ionization mode. S-O-MEL was monitored in negative ionization mode (period 1), while MEL and 6-O-MEL were detected in positive ionization mode (period 2). All calibration curves showed good linearity (r>0.991) over the concentration range with a lower limit of quantification (LLOQ) of 0.02 ng/mL for MEL, 0.04 ng/mL for 6-O-MEL and 0.50 ng/mL for S-O-MEL. The intra- and inter-day precision was within 13.5% in terms of relative standard deviation (RSD%) and the accuracy within 13.0% in terms of relative error. This convenient and specific LC-MS/MS method was successfully applied to the pharmacokinetic study of MEL and its metabolites in Beagle dogs after an oral dose of 2.0mg MEL. After ingestion of MEL, S-O-MEL was the predominant component circulating in blood. 6-O-MEL showed similar pharmacokinetic profile to that of MEL.