Nociception and hypersensitivity involve distinct neurons and molecular transducers in Drosophila.

Acute nociception is essential for survival by warning organisms against potential dangers, whereas tissue injury results in a nociceptive hypersensitivity state that is closely associated with debilitating disease conditions, such as chronic pain. Transient receptor potential (Trp) ion channels expressed in nociceptors detect noxious thermal and chemical stimuli to initiate acute nociception. The existing hypersensitivity model suggests that under tissue injury and inflammation, the same Trp channels in nociceptors are sensitized through transcriptional and posttranslational modulation, leading to nociceptive hypersensitivity. Unexpectedly and different from this model, we find that in Drosophila larvae, acute heat nociception and tissue injury-induced hypersensitivity involve distinct cellular and molecular mechanisms. Specifically, TrpA1-D in peripheral sensory neurons mediates acute heat nociception, whereas TrpA1-C in a cluster of larval brain neurons transduces the heat stimulus under the allodynia state. As a result, interfering with synaptic transmission of these brain neurons or genetic targeting of TrpA1-C blocks heat allodynia but not acute heat nociception. TrpA1-C and TrpA1-D are two splicing variants of TrpA1 channels and are coexpressed in these brain neurons. We further show that Gq-phospholipase C signaling, downstream of the proalgesic neuropeptide Tachykinin, differentially modulates these two TrpA1 isoforms in the brain neurons by selectively sensitizing heat responses of TrpA1-C but not TrpA1-D. Together, our studies provide evidence that nociception and noncaptive sensitization could be mediated by distinct sensory neurons and molecular sensors.

Quantification and discovery of quality markers from Toddalia asiatica by UHPLC-MS/MS coupled with chemometrics.

INTRODUCTION: Toddalia asiatica (TA) is a classical traditional Chinese medicine used to treat rheumatoid arthritis and contusions. However, research regarding TA quality control is currently limited. OBJECTIVE: We aimed to establish a strategy for identifying quality markers that can be used for the evaluation of the quality of TA. METHOD: A rapid and efficient ultra-high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UHPLC-MS/MS) method was developed for the quantitative determination of 19 compounds in TA from different regions. Then, the extraction process of TA was successively optimized by single-factor optimization and response surface methodology. Moreover, chemometrics was employed to confirm the correlation between quality and target compounds. RESULTS: Utilizing the UHPLC-MS/MS method, separation of the 19 bioactive compounds was achieved within 14 min. The method was validated in terms of linearity (r2 > 0.9982), precision (0.08%-3.70%), repeatability (0.50%-2.54%), stability (2.26%-5.46%), and recovery (95.8%-113%). The optimal extraction process (extraction solvent, 65% ethanol aqueous solution; solid-liquid ratio, 1:20; extraction time, 25 min) was determined with the total content of 19 bioactive compounds as indicator. Significant disparities were observed in the contents of target compounds across different batches of TA. Besides, all samples could be categorized into two distinct groups, and magnoflorine, (-)-lyoniresinol, nitidine chloride, norbraylin, skimmianine, and decarine were identified as quality markers. CONCLUSION: In the present study, we developed a strategy to improve the quality control of TA. In consideration of the pharmacodynamic activity and statistical differences, six compounds are proposed as quality markers for TA.

A novel reverse migration micellar electrokinetic chromatography method for in-capillary screening and quantifying of antioxidant components in Sanyetangzhiqing using 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) as oxidation-free radical.

A novel method of reverse migration micellar electrokinetic chromatography (RM-MEKC) using 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) as oxidation-free radical was developed to screen the major antioxidants from Sanyetangzhiqing (SYTZQ), which is a new patent drug for diabetes. For simultaneous detection and separation of 2,2'-bis (3-ethylbenzothiazoline-6-sulfonic acid) ammonium salt (ABTS+ ) and chemical components of SYTZQ, the key factors were optimized and the method was validated under the optimal conditions. All the relative standard deviations of recovery, precision, and stability were below 6.6%. Based on these, the RM-MEKC method has been successfully used to screen the main antioxidants from SYTZQ tablets. Five compounds, including rosmarinic acid, salvianolic acid B, lithospermic acid, hyperoside, and isoquercetin, were separated and identified as the major antioxidants. There was a linear relationship with correlation coefficient of 0.923 between the total amount of major antioxidants and total antioxidative activity of SYTZQ. Consequently, these five ingredients could be selected as combinatorial markers for quality control of SYTZQ, and the established method was concluded to be a simple, reliable, and powerful tool to screen and quantify active compounds for quality evaluation of SYTZQ.

Palladium-Catalyzed Sequential Heck Reactions of Olefin-Tethered Aryl Iodides with Alkenes.

An efficient approach to functionalized (E)-3-cinnamyl-3-methyl-2,3-dihydrobenzofurans and (E)-(3-methyl-2,3-dihydrobenzofuran-3-yl)but-2-enones has been developed through a Pd-catalyzed one-pot cascade process involving two sequential Heck reactions, that is, an intramolecular Heck reaction of olefin-tethered aryl iodides and an intermolecular Heck reaction with substituted styrenes and α,ß-unsaturated ketones. As a result, a series of desired products were obtained in moderate to good yields and with exclusive E-form selectivities.

Recent advances of green pretreatment techniques for quality control of natural products.

A few advancing technologies for natural product analysis have been widely proposed, which focus on decreasing energy consumption and developing an environmentally sustainable manner. These green sample pretreatment and analysis methods following the green Analytical Chemistry (GAC) criteria have the advantage of improving the strategy of chemical analyses, promoting sustainable development to analytical laboratories, and reducing the negative effects of analysis experiments on the environment. A few minimized extraction methodologies have been proposed for replacing the traditional methods in the quality evaluation of natural products, mainly including solid-phase microextraction (SPME) and liquid phase microextraction (LPME). These procedures not only have no need for large numbers of samples and toxic reagent, but also spend a small amount of extraction and analytical time. This overview aims to list out the main green strategies on the application of quality evaluation and control for natural products in the past 3 years.

Preharvest application of prohydrojasmon affects color development, phenolic metabolism, and pigment-related gene expression in red pear (Pyrus ussuriensis).

BACKGROUND: Peel color is an economically relevant trait that influences the appearance and quality of red pear, whose red color is due to anthocyanin accumulation. Prohydrojasmon (PDJ), which has similar effects to endogenous jasmonates, was developed as a commercial bioregulator, particularly to improve fruits coloring. However, little information is available about the effect of PDJ on pears. This study investigated the effects of preharvest PDJ treatments on color development, phenolic compounds accumulation, and related gene expression in the red pear cultivar 'Nanhong'. The treatments were performed during the pre-color-change period by spraying 50 or 100 mg L-1 of PDJ on fruits. RESULTS: Preharvest PDJ treatments had a significant effect on color development, without affecting other quality parameters such as total soluble solids and fruit acidity. Liquid chromatography-mass spectrometry analysis showed that concentrations of anthocyanins and flavonols were enhanced in the peel after PDJ treatments, particularly when a concentration of 100 mg L-1 was used, whereas those of hydroxycinnamates and flavanols were decreased. After PDJ application, the transcription levels of anthocyanin biosynthesis genes PAL, CHS, CHI, ANS, F3H, and UFGT were enhanced, especially under the higher PDJ concentration tested. In addition, anthocyanin accumulation in the peels of PDJ-treated fruits was found to be positively correlated with the upregulation of the regulatory gene MYB114. CONCLUSION: Preharvest treatments with PDJ could be a useful tool to improve fruits coloring and increase phenolic content in pear. These findings also improve our understanding of the molecular mechanisms associated with PDJ-regulated anthocyanin accumulation in pear fruits.

Integrated metabolomic and transcriptomic analysis reveals the effects and mechanisms of Jinqi Jiangtang tablets on type 2 diabetes.

BACKGROUND: Type 2 diabetes (T2DM) is one of the major metabolic diseases and poses a serious challenge to human life and global economic development. Jinqi Jiangtang Tablets (JQJT) is effective in ameliorating the effects of T2DM, but the mechanism of JQJT is unclear. PURPOSE: This study integrated metabolomics and transcriptomics to reveal the mechanism by which JQJT improves T2DM. METHODS: The T2DM mouse model was established, and the effects of JQJT on improving T2DM were evaluated by determining the levels of blood lipids, fasting blood glucose (FBG), insulin metabolism and hepatic lipid accumulation in mice after JQJT administration for 8 weeks. Serum metabolites were detected using ultra-performance liquid chromatography/quadrupole time-of-flight-tandem mass spectrometry (UPLC-Q-TOF-MS) technology, and mouse liver differential genes were detected using transcriptomic technology. Correlation analysis was used to extract metabolites and RNA with correlations, and potential pathways were enriched and constructed using the common pathway analysis function of MetaboAnalyst 5.0. Finally, the expression of key target proteins and genes was verified by Western blot (WB) and Polymerase Chain Reaction (PCR) to further elucidate the mechanism by which JQJT improves T2DM. RESULTS: JQJT reduced FBG and lipid levels, improved insulin resistance (IR) and hepatic lipoatrophy in mice. A total of 35 differentially abundant metabolites were identified by metabolomics, and 328 differential genes were detected by transcriptomics. The integrated metabolomics and transcriptomics results suggested that JQJT may ameliorate T2DM mainly by regulating glucose and lipid metabolic pathways. WB and PCR results showed that JQJT regulates the insulin signaling pathway, involved in fatty acid metabolism, glycogen synthesis and catabolism. CONCLUSIONS: JQJT improved IR in T2DM mice by regulating the insulin signaling pathway, improving glycogen synthesis and glycolysis, and increasing hepatic triglyceride and fatty acid metabolism.

A UPLC-MS/MS coupled with GC-MS method for quantification of twenty-one chemical ingredients from Suxiao Jiuxin pill in multiple tissue of rat and its application to tissue distribution study.

Suxiao Jiuxin pill (SJP) was a commonly-used traditional Chinese medicine for treating cardiovascular diseases. It was composed of the rhizome of Ligusticum chuanxiong Hort. and Borneolum Syntheticum. The distribution of SJP in vivo was still ambiguous. A UPLC-MS/MS coupled with GC-MS method was developed to quantify twenty-one chemical ingredients in multiple tissues from rat after administration of SJP. Protein precipitation and liquid-liquid microextraction were both utilized in sample pretreatment. All analytes were detected under acceptable specificity, linearity (correlation coefficient > 0.992), sensitivity (LLOQ < 12.5 ng/mL), precision (RSD < 14.8 %), accuracy (RE < ±14.6 %), extraction recovery (between 52.8 % and 124.1 %), matrix effect (ranged from 60.5 % and 149.7 %) and stability (RE < ±16.0 %). The established method was successfully applied in the tissue distribution study of SJP in rats. As a result, the distribution characteristics of ten analytes were clearly elucidated, including borneol, isoborneol, ligustilide, senkyunolide A, ferulic acid, senkyunolide I, levistolide A, neocnidilide, senkyunolide H and angelicide. The information provided by this research was greatly meaningful for the active chemical ingredient exploration and clinical application of SJP.

Determination of eleven components in rat plasma by UPLC-MS/MS and GC-MS for pharmacokinetic studies after oral administration of Citri Reticulatae Pericarpium extract.

Citri Reticulatae Pericarpium (CRP) is used as common health-care food and traditional Chinese medicine (TCM), which exerts pharmacological effects, such as anti-cardiovascular, anti-tumor, anti-oxidant, anti-inflammatory, anti-virus, hepatoprotective, blood pressure-lowering and neuroprotective. In this study, reliable, and sensitive ultra-high performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) and gas chromatography-mass spectrometry (GC-MS) methods were developed and validated for the determination of eleven active components in rat plasma after oral administration of the CRP extract. The results of this method exhibited that the specificity, linearity (r > 0.999), precision and accuracy (the coefficient of variation (CV) < 11.5 %), recovery (52.9-107.9 %), matrix effects (63.8-107.5 %), and stability (CV < 10.8 %) met all requirements for the quantitation of plasma samples. The pharmacokinetic results showed that the Tmax of flavone glycosides was less than 0.7 h, and that of polymethoxyflavones and volatile components were within 1-7 h. Meanwhile, the area-under-the-curve (AUC) and concentration maximum (Cmax) of hesperidin, nobiletin, tangeretin, and D-limonene were higher than those of the other components, suggesting that the plasma exposure levels of these constituents were higher in CRP. The present research lays a foundation for elucidating the therapeutic material basis and provides a reference for further scientific research and clinical application of CRP.

Investigation of the drug-drug interaction and incompatibility mechanism between Aconitum carmichaelii Debx and Pinellia ternata (Thunb.) Breit.

ETHNOPHARMACOLOGICAL RELEVANCE: The combination of Aconitum carmichaelii Debx (Chuanwu, CW) and Pinellia ternata (Thunb.) Breit (Banxia, BX) forms an herbal pair within the eighteen incompatible medicaments (EIM), indicating that BX and CW are incompatible. However, the scientific understanding of this incompatibility mechanism, especially the corresponding drug-drug interaction (DDI), remains complex and unclear. AIM OF THE STUDY: This study aims to explain the DDI and potential incompatibility mechanism between CW and BX based on pharmacokinetics and cocktail approach. MATERIALS AND METHODS: Ultraperformance liquid chromatography-tandem mass spectrometry methods were established for pharmacokinetics and cocktail studies. To explore the DDI between BX and CW, in the pharmacokinetics study, 10 compounds were determined in rat plasma after administering CW and BX-CW herbal pair extracts. In the cocktail assay, the pharmacokinetic parameters of five probe substrates were utilized to assess the influence of BX on cytochrome P450 (CYP) isoenzyme (dapsone for CYP3A4, phenacetin for CYP1A2, dextromethorphan for CYP2D6, tolbutamide for CYP2C9, and omeprazole for CYP2C19). Finally, the DDI and incompatibility mechanism of CW and BX were integrated to explain the rationality of EIM theory. RESULTS: BX not only enhances the absorption of aconitine and benzoylaconine but also accelerates the metabolism of mesaconitine, benzoylmesaconine, songorine, and fuziline. Moreover, BX affects the activity of CYP enzymes, which regulate the metabolism of toxic compounds. CONCLUSIONS: BX altered the activity of CYP enzymes, consequently affecting the metabolism of toxic compounds from CW. This incompatibility mechanism may be related to the increased absorption of these toxic compounds in vivo.

Simultaneous determination of multiple components in rat plasma by UHPLC-sMRM for pharmacokinetic studies after oral administration of Qingjin Yiqi Granules.

As a Traditional Chinese Medicine prescription, Qingjin Yiqi Granules (QJYQ) provides an effective treatment for patients recovering from COVID-19. However, the pharmacokinetics characteristics of the main components of QJYQ in vivo are still unknown. An efficacious ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was developed and validated for the simultaneous determination of 33 components in rat plasma after oral administration of QJYQ. The plasma samples were precipitated with 400 µL methanol/acetonitrile (1/1, v/v) and analyzed in scheduled multiple reaction monitoring mode. The linear relationship of the 33 components was good (r > 0.9928). The lower limit of quantification for 33 components ranged from 0.4-60.5 ng/mL. The average recoveries and matrix effects of the analytes ranged from 72.9% to 115.0% with RSD of 1.4%-15.0%. All inter-day and intra-day RSDs were within 15.0%. After oral administration (3.15 g/kg), the validated approach was effectively applied to the pharmacokinetics of main components of QJYQ. Finally, fifteen main constituents of QJYQ with large plasma exposure were obtained, including baicalin, wogonoside, wogonin, apigenin-7-O-glucuronide, verbenalin, isoferulic acid, hesperidin, liquiritin, harpagide, protocatechuic acid, p-Coumaric acid, ferulic acid, sinapic acid, liquiritin apioside and glycyrrhizic acid. The present research lays a foundation for clarifying the therapeutic material basis of QJYQ and provides a reference for further scientific research and clinical application of QJYQ.

Simultaneous Separation and Determination of Nine Active Ingredients in Sanyetangzhiqing by Cyclodextrin-Modified Micellar Electrokinetic Capillary Electrophoresis-Diode Array Detector.

A simple and sensitive strategy using cyclodextrin-modified micellar electrokinetic chromatography with diode array detector was developed and applied for the simultaneous separation and determination of nine components in Sanyetangzhiqing (SYTZQ), a hypoglycemic and hypolipidemic agent. Several important parameters affecting separation performance were evaluated and optimized using single variable methods. Under the optimal conditions, baseline separation of the nine components, including four flavonoids (hyperoside, isoquercitrin, quercetin-3-O-glucuronoside, and astragalin), four phenolic acids (chlorogenic acid, rosmarinic acid, salvianolic acid B, and lithospermic acid), and a monoterpenoids (paeoniflorin), were achieved in less than 16 min. The correlation coefficients of the calibration curves were over 0.9996 for all the analytes. Intraday and interday precisions ranged from 0.4% to 4.8% and 1.7% to 5.0%, respectively. Recoveries of analytes varied from 95.3% to 105%. Validation results as well as the application to analyse SYTZQ samples demonstrated the applicability of the proposed method and thus provided an effective tool for the quality control of SYTZQ. Moreover, with the advantages of short time consuming, low energy consumption, high efficiency, and low cost, this method has laid a foundation for the determination and quality evaluation of multicomponents in Chinese herbal compounds.

A comprehensive review of the botany, ethnopharmacology, phytochemistry, pharmacology, toxicity and quality control of Perillae Fructus.

ETHNOPHARMACOLOGICAL RELEVANCE: Perilla frutescens (Linnaeus) Britton, Mem. Torrey Bot. Club 5: 277. 1894., is famous as a worldwide plant with multiple medical parts, including leaves, stems, fruits, etc. Perillae Fructus, the desiccative ripe fruit of P. frutescens, is locally called Zisuzi in Chinese Pharmacopoeia. It is a popularly used herb for relieving cough and asthma, dissipating phlegm and treating constipation in some Asian countries, such as China, Japan, India, South Korea, etc. Various chemical compounds were isolated and identified from Perillae Fructus. THE AIM OF THE REVIEW: This review aims to summarize the botany, ethnopharmacological applications, phytochemistry, pharmacology, toxicity and quality control of Perillae Fructus to provide scientific evidence for development and utilization Perillae Fructus. MATERIALS AND METHODS: Relevant information about Perillae Fructus was collected from ScienceDirect, PubMed, Web of science, CNKI, WanFang data, ancient classics and clinical reports. Some electronic databases were also retrieved. RESULTS: Perillae Fructus was exerted to treat cough and asthma in traditional application. It also had the effect on moistening intestine to relieve constipation for tremendous lipid substances. Up to now, 193 compounds have been isolated and identified from Perillae Fructus, mainly including fatty acids, flavonoids, phenolic acids, phytosterols, triterpenoids and volatile oils. As for its pharmacological activities, prevalent traditional applications of Perillae Fructus have been supported by modern pharmacological experiments in vivo or in vitro, such as anti-inflammatory and anti-oxidant effects. Besides, Perillae Fructus also has hypolipidemic, anti-tumor, antibacterial effects, etc. This review will provide a scientific basis for further studies and rational applications of Perillae Fructus in the future. CONCLUSIONS: According to its traditional applications, phytochemicals and pharmacological activities, Perillae Fructus was regarded as a valuable herb for application in medicine and food fields. Although some ingredients have been confirmed to have multiple pharmacological activities, their mechanisms of action are still unclear. Further studies on the material basis and mechanism of action are clearly warranted.

A Non-Immunized and BSA-Template Aggregation-Induced Emission Sensor for Noninvasive Detection of Cystatin C in the Clinical Diagnosis of Diabetes Nephropathy.

Diabetes nephropathy (DN) is one of the main causes of death in patients with diabetes. Cystatin C (Cys C) is a reliable indicator of glomerular filtration function. Therefore, it is urgent and meaningful to obtain early warning of DN by noninvasive measurement of Cys C. In this investigation, a novel fluorescence sensor (BSA-AIEgen sensor) was synthesized by cross-linking the aggregation-induced emission (AIE) characteristics of 2-(4-bromophenyl)-3-(4-(4-(diphenylamino) styryl) phenyl) fumaronitrile (TPABDFN) and bovine serum albumin (BSA), which exhibited the "On" state owing to the restriction of the intramolecular motions (RIM) phenomenon of TPABDFN. Intriguingly, a decrease in fluorescence of BSA-AIEgen sensors could be found owing to BSA on the surface of BSA-AIEgen sensor hydrolyzed by papain, but a reverse phenomenon emerged with the increase of Cys C content as the inhibitor of papain. Hence, Cys C was successfully detected by employing the fluorescent differential display and the linear range was from 12.5 ng/mL to 800 ng/mL (R2 = 0.994) with the limit of detection (LOD) of 7.10 ng/mL (S/N = 3). Further, the developed BSA-AIEgen sensor successfully differentiates patients with diabetes nephropathy from volunteers with the advantages of high specificity, low cost, and simple operation. Accordingly, it is expected to become a non-immunized method to monitor Cys C for the early warning, noninvasive diagnosis, and drug efficacy evaluation of diabetes nephropathy.

An efficient method for qualitation and quantitation of multi-components of the herbal medicine Qingjin Yiqi Granules.

Qingjin Yiqi Granules (QJYQ) is a Traditional Chinese Medicines (TCMs) prescription for the patients with post-COVID-19 condition. It is essential to carry out the quality evaluation of QJYQ. A comprehensive investigation was conducted by establishing deep-learning assisted mass defect filter (deep-learning MDF) mode for qualitative analysis, ultra-high performance liquid chromatography and scheduled multiple reaction monitoring method (UHPLC-sMRM) for precise quantitation to evaluate the quality of QJYQ. Firstly, a deep-learning MDF was used to classify and characterize the whole phytochemical components of QJYQ based on the mass spectrum (MS) data of ultra-high performance liquid chromatography quadrupole time of flight tandem mass spectrometry (UHPLC-Q-TOF/MS). Secondly, the highly sensitive UHPLC-sMRM data-acquisition method was established to quantify the multi-ingredients of QJYQ. Totally, nine major types of phytochemical compounds in QJYQ were intelligently classified and 163 phytochemicals were initially identified. Furthermore, fifty components were rapidly quantified. The comprehensive evaluation strategy established in this study would provide an effective tool for accurately evaluating the quality of QJYQ as a whole.

Gliotransmission and adenosine signaling promote axon regeneration.

How glia control axon regeneration remains incompletely understood. Here, we investigate glial regulation of regenerative ability differences of closely related Drosophila larval sensory neuron subtypes. Axotomy elicits Ca2+ signals in ensheathing glia, which activates regenerative neurons through the gliotransmitter adenosine and mounts axon regenerative programs. However, non-regenerative neurons do not respond to glial stimulation or adenosine. Such neuronal subtype-specific responses result from specific expressions of adenosine receptors in regenerative neurons. Disrupting gliotransmission impedes axon regeneration of regenerative neurons, and ectopic adenosine receptor expression in non-regenerative neurons suffices to activate regenerative programs and induce axon regeneration. Furthermore, stimulating gliotransmission or activating the mammalian ortholog of Drosophila adenosine receptors in retinal ganglion cells (RGCs) promotes axon regrowth after optic nerve crush in adult mice. Altogether, our findings demonstrate that gliotransmission orchestrates neuronal subtype-specific axon regeneration in Drosophila and suggest that targeting gliotransmission or adenosine signaling is a strategy for mammalian central nervous system repair.

A novel biological sources consistency evaluation method reveals high level of biodiversity within wild natural medicine: A case study of Amynthas earthworms as "Guang Dilong".

For wild natural medicine, unanticipated biodiversity as species or varieties with similar morphological characteristics and sympatric distribution may co-exist in a single batch of medical materials, which affects the efficacy and safety of clinical medication. DNA barcoding as an effective species identification tool is limited by its low sample throughput nature. In this study, combining DNA mini-barcode, DNA metabarcoding and species delimitation method, a novel biological sources consistency evaluation strategy was proposed, and high level of interspecific and intraspecific variations were observed and validated among 5376 Amynthas samples from 19 sampling points regarded as "Guang Dilong" and 25 batches of proprietary Chinese medicines. Besides Amynthas aspergillum as the authentic source, 8 other Molecular Operational Taxonomic Units (MOTUs) were elucidated. Significantly, even the subgroups within A. aspergillum revealed here differ significantly on chemical compositions and biological activity. Fortunately, this biodiversity could be controlled when the collection was limited to designated areas, as proved by 2796 "decoction pieces" samples. This batch biological identification method should be introduced as a novel concept regarding natural medicine quality control, and to offer guidelines for in-situ conservation and breeding bases construction of wild natural medicine.

Structural polymorphism of human telomere G-quadruplex induced by a pyridyl carboxamide molecule.

In this work, we described a kinetically slow (hour-scale) but thermodynamically favored G-quadruplex conversion induced by a pyridyl carboxamide molecule. This slow transition was observed through CD spectra and gels, and its final stable parallel conformation was identified by 2-aminopurine experiments. Kinetic experiments indicated that this slow process was a first-order reaction, implying it was a unimolecular conversion. Quite distinctly from other reported ligand-driven G-quadruplex conformation alteration, this slow conversion reveals a novel insight into G-quadruplex polymorphism, in accordance with the behavior of human telomere G-quadruplex in a molecular crowding environment in K(+) solution, further enriching the known structural polymorphism of human telomere DNA and providing new consideration for drug design based on G-quadruplexes.

The miR-410-5p /ITGA6 axis participates in the pathogenesis of recurrent abortion by regulating the biological function of trophoblast.

The purpose of this study was to determine the regulation of the miR-410-5p /ITGA6 axis on the biological functions of trophoblast cells and the mechanism involved in recurrent spontaneous abortion（RSA）. We used qRT-PCR and Western blotting to quantify the expression levels of Mir-410-5p and ITGA6 in placenta of RSA and normal, and found that compared with normal placenta, the placenta of RSA patients showed higher miR-410-5p and lower ITGA6 expression. Dual luciferase reporter gene assay confirmed the binding of miR-410-5p to ITGA6. The expression of miR-410-5p and ITGA6, and proliferation, apoptosis, invasion and migration of trophoblast cells and the effect on the polarization of M2 macrophages were detected in the trophoblast derived cell lines HTR8/Svneo transfected with miR-410-5p mimic, sh-miR-410-5p and si-ITGA6 respectively. Meanwhile, the molecular mechanism of ITGA6 regulation on trophoblast cells was explored. Transfection with miR-410-5p mimic or si-ITGA6 attenuated the proliferation, migration and invasion and induced apoptosis of HTR-8/SVneo cells. Transfection of sh-miR-410-5p promoted proliferation, migration and invasion, and weakened apoptosis of HTR-8/SVneo cells. In addition, overexpression of miR-410-5p in trophoblast cells inhibited the polarization of M2 macrophages, while knockdown of miR-410-5p was beneficial to recruitment of trophoblast cell and promoted the polarization of M2 macrophages. ITGA6 may affect the biological functions of trophoblast cells by regulating PI3K/AKT and MAPK signaling pathways. In conclusion, miR-410-5p mediates trophoblast cell proliferation, apoptosis, invasion and migration through regulating ITGA6 expression.

An online stepwise background subtraction-based ultra-high pressure liquid chromatography quadrupole time of flight tandem mass spectrometry dynamic detection integrated with metabolic molecular network strategy for intelligent characterization of the absorbed chemical-fingerprint of QiangHuoShengShi decoction in vivo.

QiangHuoShengShi decoction (QHSS) was an ancient and classical traditional Chinese medicine (TCM) prescription. In the previous study, its phytochemical fingerprint had been comprehensively characterized. However, no reports were available on its absorbed prototypes and the related metabolites in rat plasma samples. In this study, an intelligent and innovate analysis strategy was built for characterizing metabolic chemical-fingerprint in rat plasma after oral administration of QHSS extract. Firstly, a very simple and highly efficient online stepwise background subtraction (BS)-based ultra-high pressure liquid chromatography quadrupole time of flight tandem mass spectrometry (UHPLC-Q-TOF-MS/MS) dynamic detection method was established to analyze the plasma samples. Secondly, the intelligent metabolic molecular network (MMN) technology was developed and used for rapidly screening out the metabolites of interest, which was followed by prediction of chemical types using the modified deep-learning assisted mass defect filter (MDF) analysis. Thirdly, the screened metabolites with identification features (metabolic pathways and chemical classification) were deeply characterized based on the MS/MS datasets. Finally, 58 prototypes of QHSS were successfully acquired and subsequently identified, including coumarins, chromones, phthalides, phenolic acids, flavonoids, and saponins. A total of 111 metabolites of the coumarins, chromones, phthalides were filtered to be tentatively characterized. This developed qualitative strategy was very helpful to quickly target medicine-related metabolites in the complex bio-matrix and, importantly, it could further visualize medicine-metabolic pathways hidden in the messy mass spectrum datasets. In all, the innovate strategy would provide a powerful tool for effectively acquiring and decode complex metabolic fingerprint of natural products in vivo.