A simple and sensitive HPLC-MS/MS assay for the quantitation of montelukast in cell-based systems in vitro pulmonary drug permeability study.

Montelukast is a potent and selective antagonist of the cysteinyl leukotriene receptor 1 subtype (CysLT1) and widely used in the form of oral tablets and granules for asthma prophylaxis and treatment. Recently, due to the pulmonary inhaled administration can limit montelukast distribution in the systemic circulation, avoid the first-pass metabolism and have better therapeutic effects in respiratory disease treatment, explore alternative routes of administration, like delivery of montelukast via an inhaled, is a new research trend for montelukast. The aim of the current study was to develop and validate a simple, accurate, highly sensitive and selective liquid chromatography-tandem mass spectrometry method (LC-MS/MS) for determination of montelukast in an in vitro cell-based pulmonary pharmacokinetics system model, which can be used to be a better understanding the fate of inhaled montelukast in the lungs. In this study, montelukast was extracted by protein precipitation with acetonitrile containing labeled montelukast. The chromatography was performed on an Agilent Eclipse plus C8 column (4.6 mm × 100 mm, 3.5 µm, Darmstadt, Germany) operating at 35 ◦C. The mobile phase consisted of acetonitrile: 20 mM ammonium formate buffer (80: 20, v/v), was delivered at a flow rate of 0.5 mL/min. montelukast and the internal standard were both eluted at 4.2 min. A linear (1/x2) relationship was used to perform the calibration over an analytical range from 0.5 to 600 ng/mL. The intra- and inter-batch precision expressed as CV for four QC samples including LLOQ range from 1.14 % to 6.25 %. The intra- and inter-batch accuracy for four concentrations of montelukast were in the range of 95.19%-104.1%. All the values for accuracy and precision were within the acceptance range. The method met all the bioanalytical method validation requirements by ICH and was suitable for the assay of montelukast which in the in vitro cell-based pulmonary pharmacokinetics system model.

Discovery of the possible mechanisms in kouyanqing granule for treatment of oral ulcers based on network pharmacology.

BACKGROUND: Oral ulcer diseases are complex inflammatory diseases caused by multi-factors, which severely impact patient quality of life. Kouyanqing Granule (KYQG) has been used to treat inflammatory diseases of the mouth and throat, including recurrent aphthous stomatitis (RAS), traumatic ulcers, oral leukoplakia and so on. However, the underlying molecular mechanisms of KYQG in treating these diseases are still unclear. We aimed to explore the possible mechanisms in KYQG for the treatment of oral ulcers. METHODS: An innovative network pharmacology method was established by incorporating targets searching and fishing, network analysis, and silico validation to discover the pharmacological mechanisms of action of KYQG for the treatment of oral ulcers. Then, we verified the reliability of this method by an animal experiment. RESULTS: Our data indicated that a total of 47 key targets were screened, which mainly involved in three function modules including the inhibition of inflammation, the regulation of immunological response, and the suppression of oxidative stress. The implementation of these functions relies on the complex multi-pathways network, especially TNF signaling pathway and HIF-1 signaling pathway. The results of the experimental verification indicated that KYQG significantly inhibited the serum levels of cyclooxygenase-2 (COX2), matrix metalloproteinase 9 (MMP9) and tumor necrosis factor-alpha (TNF-α) in rats with oral ulcer. CONCLUSION: KYQG exhibited the therapeutic effects on oral ulcers probably by inhibiting inflammation, regulating immunological response, and suppressing oxidative stress through a complex multi-pathways network. Particularly, TNF signaling pathway and HIF-1 signaling pathway may play crucial roles in the protection of KYQG against oral ulcers. This work not only offers a method for understanding the functional mechanisms of KYQG for treating oral ulcer diseases from a multi-scale perspective but also may provide an efficient way for research and development of complex composition formula.

Ferroelectric Enhanced Performance of a GeSn/Ge Dual-Nanowire Photodetector.

GeSn offers a reduced bandgap than Ge and has been utilized in Si-based infrared photodetectors with an extended cutoff wavelength. However, the traditional GeSn/Ge heterostructure usually consists of defects like misfit dislocations due to the lattice mismatch issue. The defects with the large feature size of a photodetector fabricated on bulk GeSn/Ge heterostructures induce a considerable dark current. Here, we demonstrate a flexible GeSn/Ge dual-nanowire (NW) structure, in which the strain relaxation is achieved by the elastic deformation without introducing defects, and the feature dimension is naturally at the nanoscale. A photodetector with a low dark current can be built on a GeSn/Ge dual-NW, which exhibits an extended detection wavelength beyond 2 µm and enhanced responsivity compared to the Ge NW. Moreover, the dark current can be further suppressed by the depletion effect from the ferroelectric polymer side gate. Our work suggests the flexible GeSn/Ge dual-NW may open an avenue for Si-compatible optoelectronic circuits operating in the short-wavelength infrared range.

Effects of Total Flavonoids from Exocarpium Citri Grandis on Air Pollution Particle-Induced Pulmonary Inflammation and Oxidative Stress in Mice.

Exocarpium Citri Grandis (ECG) is a famous traditional Chinese medicine, which has been commonly used to alleviate cough and phlegm for more than several hundred years, and total flavonoids are the main effective components of this medicine. This study investigated the effects of total flavonoids from ECG (TFECG) on pulmonary inflammation and oxidative stress induced by PM2.5 in mice. Model mice received an intratracheal instillation of PM2.5 (10 mg/mL) once at day 0. Bronchoalveolar lavage fluid (BALF) was collected after 72 hr to measure the total number of white blood cell (WBC), neutrophils (NEUT), lymphocytes (LYMPH), and monocytes (MONO). The levels of tumor necrosis factor-α (TNF-α), interleukin 1ß (IL-1ß), interleukin-6 (IL-6), and interleukin-18 (IL-18) in BALF were quantified by using enzyme-linked immunosorbent assay kits. Lung tissues were used to determine the contents of total protein (TP), malondialdehyde (MDA), nitric oxide (NO), reduced glutathione (GSH), oxidized glutathione (GSSG) and the activities of superoxide dismutase (SOD), lactate dehydrogenase (LDH), Glutathione Peroxidase (GSH-Px), and inducible nitric oxide synthase (iNOS). We found that TFECG significantly inhibited PM2.5 -stimulated overproduction of TNF-α, IL-1ß, IL-6, and IL-18 and increased the numbers of WBC, NEUT, LYMPH, and MONO in BALF. TFECG observably relieved the PM2.5 -induced increases in the contents of TP, MDA, and NO, and the activities of LDH and iNOS. TFECG also alleviated PM2.5 -induced decreases in the activities of iNOS and GSH-Px as well as GSH/GSSG ratio. The results indicate that TFECG has anti-inflammatory and antioxidant activities, which may potentially contribute to the treatment of PM2.5 -induced lung injury. PRACTICAL APPLICATION: Exocarpium Citri Grandis (ECG) is rich in flavonoids, which are beneficial to improve anti-inflammation and antioxidant capacity. We proved that total flavonoids of ECG had a positive therapeutic effect on PM2.5 -induced lung injury, which expands the potential applications of ECG in the dietary supplement industries.

Pharmacodynamic effects of Dan-hong injection in rats with blood stasis syndrome.

Dan-hong injection (DHI) is extracted from Salvia miltiorrhiza (SM) and Carthamus tinctorius (CT) and is widely used for the treatment of cardiovascular diseases. Our previous results showed DHI could improve hemorheology in rats. Since complex cellular interactions such as inflammation and oxidative stress are believed to be implicated in the pathogenesis of cardiovascular events, investigation of such pathological factors will contribute substantially to the understanding of the features and mechanisms of DHI. Therefore, in this study we used a rat model of blood stasis to explore the overall effects of DHI by detecting twenty three indexes, which were related to inflammation, immune response, vascular endothelial function, myocardial energy metabolism, oxidative stress, platelet aggregation, liver and renal function. Meanwhile, the interaction between SM and CT was discussed by comparing the effects of each single herb. DHI could significantly decrease the serum contents of IL-1ß, TNF-α, IL-6, IL-8, IgM, IgG, IgA, MPO, hs-CRP, MDA, LDH, CK-MB, PAF, ALP and Cr, while elevate NO, SOD, TP and UA levels, indicating that DHI could inhibit inflammation and platelet aggregation, thereby relieving immune response and peroxidation, protecting vascular endothelial and organ function, and then prevent and treat cardiovascular diseases. In terms of compatibility, SM and CT showed complementary effects on markers of inflammatory and oxidative status, vascular endothelial damage and myocardial energy metabolism. On the other hand, they counteracted each other and SM reduced the side effects of creatinine caused by CT. This study contributes to comprehensively understand the pharmacodynamics effects and mechanism of DHI.

NO2 gas sensor based on graphene decorated with Ge quantum dots.

We report a NO2 gas sensor based on germanium quantum dots (GeQDs)/graphene hybrids. Graphene was directly grown on germanium through chemical vapor deposition and the GeQDs were synthesized via molecular beam epitaxy. The samples were characterized by atomic force microscope, Raman spectra, scanning electron microscope, x-ray photoelectron spectroscope and transmission electron microscope with energy dispersive x-ray. By introducing GeQDs on graphene, the gas sensor sensitivity to NO2 was improved substantially. With the optimization of the growth time of GeQDs (600 s), the response sensitivity to 10 ppm NO2 can be as high as 3.88, which is 20 times higher than that of the graphene sensor without GeQDs decoration. In addition, the 600 s GeQDs/graphene hybrid sensor exhibits fast response and recovery rates as well as excellent stability. Our work may provide a new route to produce low-power consumption, portable, and room temperature gas sensor which is amenable to mass production.

UFLC-Q-TOF-MS/MS-Based Screening and Identification of Flavonoids and Derived Metabolites in Human Urine after Oral Administration of Exocarpium Citri Grandis Extract.

Exocarpium Citri grandis (ECG) is an important Traditional Chinese Medicine (TCM) for the treatment of cough and phlegm, and the flavonoids contained were considered the main effective components. To date, the systematic chemical profiling of these flavonoids and derived in vivo metabolites in human have not been well investigated. ECG was extracted using boiling water and then provided to volunteers for oral administration. Following the ingestion, urine samples were collected from volunteers over 48 h. The extract and urine samples were analyzed using ultra-fast liquid chromatography/quadrupole-time-of-flight tandem mass spectrometry (UFLC-Q-TOF-MS/MS) system to screen and identify flavonoids and derived in vivo metabolites. A total of 18 flavonoids were identified in the ECG extract, and 20 metabolites, mainly glucuronide and sulfate conjugates, were screened in urine samples collected post consumption. The overall excretion of naringenin metabolites corresponded to 5.45% of intake and occurred mainly within 4-12 h after the ingestion. Meanwhile, another 29 phenolic catabolites were detected in urine. Obtained data revealed that flavonoids were abundant in the ECG extract, and these components underwent extensive phase II metabolism in humans. These results provided valuable information for further study of the pharmacology and mechanism of action of ECG.

Identification and Comparison of Constituents of Aurantii Fructus and Aurantii Fructus Immaturus by UFLC-DAD-Triple TOF-MS/MS.

Although Aurantii Fructus (AF) and Aurantii Fructus Immaturus (AFI) are both the fruits of the same rutaceae plant at different stages of growth, they exert similar yet distinct clinical effects. The chemical composition is crucial for quality control as well as therapeutic application. To address this concern, it is significant to evaluate the similarities and differences of the constituents in both AF and AFI. The extract of AF and AFI were comprehensively analyzed by ultra fast liquid chromatography-photodiode array detector-triple-time of flight-tandem mass spectrometry (UFLC-DAD-Triple TOF-MS/MS). Among the 40 compounds detected, 19 metabolites were detected in both the AF and AFI; whereas 13 compounds were only detected in AF and five constituents were exclusively detected in AFI. In particular, even in AFI, three compounds were only identified in AFI (Citrus aurantium' L. and its cultivar). Among the 18 compounds confirmed by standard database, 13 compounds were reported in AF and AFI for the first time. Furthermore, the distinction was also revealed by the content of naringin, hesperidin, neohesperidin, and synephrine. The study directly contributed to the similarities and differences of AF and AFI. Herein, similarities and the differences in chemical profiles of AF and AFI could explain the current clinical applications.

Comprehensive investigation into the interconversion of C-2 diastereomers of naringin.

Naringin is a flavanone that widely presents on daily diet and traditional medicinal materials. The ratios of naringin C-2 diastereomers are found different in reported samples, thus suspiciously leading to various functions. In this study, we measured the interconversion of C-2 diastereomers intensively with ultimate high-performance liquid chromatography and circular dichroism spectra. We examined the diastereomeric naringins in fresh citrus fruit, Huajuhong decoction pieces, and naringin tablet; evaluated the impact of tablet production procedures in factory; and monitored the rapid racemization in incubation. The results not only confirmed that enzyme, temperature, and pH condition could influence the interconversion but also demonstrated that diverse ratios of diastereomers showed limited influence on metabolic behaviors of naringin in the blood, which consequently cause comparable bioactivities. This study could provide comprehensive understanding of diastereomeric interconversion and provide useful reference for drug development.

Toward a scientific understanding of the effectiveness, material basis and prescription compatibility of a Chinese herbal formula Dan-hong injection.

Since traditional Chinese medicine (TCM) is a complex mixture of multiple components, the application of methodologies for evaluating single-components Western medicine in TCM studies may have certain limitations. Appropriate strategies that recognize the integrality of TCM and connect to TCM theories remain to be developed. Here we use multiple unique approaches to study the scientific connotation of a TCM formula Dan-hong injection (DHI) without undermining its prescription integrity. The blood circulation improving and healing promoting effects of DHI were assessed by a qi stagnation blood stasis rat model and a mouse model of laser irradiation induced cerebral microvascular thrombosis. By UFLC-PDA-Triple Q-TOF-MS/MS and relevance analysis between chemical characters and biological effects, 82 chemical constituents and nine core components, whose blood circulation promoting effects were found comparable to that of whole DHI, were successfully identified. What's more, the rationality of DHI prescription compatibility could be reflected not only in the maximum efficacy of the original ratio, but also in the interactions of compounds from different ingredient herbs, such as complementary activities and facilitating tissues distribution. This study provides scientific evidences in explanation of the clinical benefits of DHI, and also gives a good demonstration for the comprehensive evaluation of other TCM.

[Effects of ozone stress on photosynthesis and dry matter production of rice II -you 084 under different Planting densities].

In order to investigate the effects of ozone stress on photosynthesis, dry matter production, non-structural carbohydrate and yield formation of rice, a free air ozone concentration enrichment (FACE) experiment was conducted. A super hybrid rice cultivar II-you 084 with 3 spacing levels, low plant density (LD, 16 hills per m2), medium (MD, 24 hills per m2) and high plant density (HD, 32 hills per m2), was grown in the field at current and elevated ozone concentrations (current × 1.5). The results were as follows: Elevated ozone significantly reduced leaf SPAD value of UI-you 084 by 6%, 11% and 13%, at 63, 77, and 86 days after transplanting, respectively. The declines in leaf net photosynthetic rate, stomatal conductance and transpiration rate at filling stage increased significantly on ozone stress over time. Ozone stress decreased dry matter production of rice by 46% from heading stage to plant maturity, thus reduced biomass yield by 25%. Elevated ozone decreased the concentration and accumulation of soluble carbohydrate and starch in stem of II-you 084 at jointing, heading and plant maturity, but significantly increased the dry matter transportation rate. No significant interaction was observed between ozone and planting density for photosynthesis, dry matter production and non-structural carbohydrate of rice. The above results indicated that elevated ozone reduced photosynthesis and growth of rice II-you 084 at late growth stage, which had no relationship with planting density.

Identification and pharmacokinetics of multiple potential bioactive constituents after oral administration of radix astragali on cyclophosphamide-induced immunosuppression in Balb/c mice.

Radix Astragali (RA) is one of the commonly-used traditional Chinese medicines (TCMs) with an immunomodulatory effect confirmed in the clinic. In order to better understand the material basis for the therapeutic effects, this study was to investigate the absorbed components and their pharmacokinetic profile after oral administration of RA on cyclophosphamide-induced immunosuppression in Balb/c mice. As a result, 51 compounds in RA extract and 31 prototype compounds with nine metabolites were detected in mice plasma by the ultra-fast liquid chromatography (UFLC)-DAD-Q-TOF-MS/MS method. The pharmacokinetic parameters of five main constituents, including calycosin-7-O-glucoside, ononin, calycosin, formononetin and astragaloside IV, were obtained using HPLC-MS/MS. These results offered useful information for research on the pharmacological mechanism of RA and for its further development.

Improved UV photoresponse of ZnO nanorod arrays by resonant coupling with surface plasmons of Al nanoparticles.

In this study, localized surface plasmon resonance mediated by aluminium nanoparticles (Al NPs) was employed to enhance the ultraviolet (UV) response of ZnO nanorod array (NRA) photodetectors grown vertically on a Quartz substrate using a simple vapor transport method. The responsivity of the ZnO NRA photodetector decorated with Al NPs was enhanced from 0.12 to 1.59 A W(-1) and the sensitivity and response rate have been improved greatly compared with those of the bare one. The measurement results in the transmittance spectra and time-resolved photoluminescence spectra suggest that the improved photoresponse and the enhanced spontaneous emission of the ZnO NRA photodetector with Al NP decoration are both attributed to resonant coupling between the excitons in ZnO and the localized surface plasmons (LSPs) in the Al NPs. Our results demonstrated that the plasmon-enhanced ZnO NRA photodetector has a great potential for application in building sensors with a fast response and reset time, high sensitivity, and good signal-to-noise ratio for photoelectric sensing.

Rapid Identification and Simultaneous Quantification of Multiple Constituents in Nao-Shuan-Tong Capsule by Ultra-Fast Liquid Chromatography/Diode-Array Detector/Quadrupole Time-of-Flight Tandem Mass Spectrometry.

A rapid and high-sensitive ultra-fast liquid chromatography coupled with a diode-array detector and a quadrupole/time-of-flight mass spectrometry (MS) method was established and validated for the chemical profiling of Nao-shuan-tong capsule (NSTC) and simultaneous quantification of five major constituents. A total of 59 components including monoterpene glycosides, flavonoids, sesquiterpenoids, ketosteroids, thiophenes, organic acids and alkaloids were identified or tentatively characterized in NSTC based on the accurate mass and tandem MS behavior. Five major bioactive constituents were chosen as the chemical indexes of holistic quality evaluation and quantified simultaneously. All calibration curves showed good linear regression (r(2) > 0.9991) in the range 25.2-510, 145-2,900, 1.84-36.8, 2.61-52.2 and 3.25-26.2 µg/mL for gastrodin, paeoniflorin, typhaneoside, ß-ecdysterone and isorhamnetin-3-O-neohesperidoside, respectively. It also showed good precision, stability and accuracy for quantification of these five compounds. The limit of detections and limit of quantitations for the analytes ranged from 0.14 to 1.09 µg/mL and from 0.47 to 3.63 µg/mL, respectively. The validated quantification method was applied to analyze 10 batches of commercial NSTC. These results will provide a basis for quality control of the production process and the further pharmacological study in vivo of NSTC.

Systematic chemical profiling of Citrus grandis 'Tomentosa' by ultra-fast liquid chromatography/diode-array detector/quadrupole time-of-flight tandem mass spectrometry.

Citrus grandis 'Tomentosa', as the original plant of the traditional Chinese medicine "Huajuhong", has been used as antitussive and expectorant in clinic for thousands of years. The fruit epicarp and whole fruit of this plant were both literarily recorded and commonly used. In the present study, an ultra-fast liquid chromatography coupled with diode-array detection and quadrupole/time-of-flight mass spectrometry (UFLC-DAD-Q-TOF-MS/MS) based chemical profiling method was developed for rapid holistic quality evaluation of C. grandis 'Tomentosa', which laid basis for chemical comparison of two medicinal parts. As a result, forty-eight constituents, mainly belonging to flavonoids and coumarins, were unambiguously identified by comparison with reference standards and/or tentatively characterized by elucidating UV spectra, quasi-molecular ions and fragment ions referring to information available in literature. Both of the epicarp and whole fruit samples were rich in flavonoids and coumarins, but major flavonoids contents in whole fruit were significantly higher than in epicarp (P<0.5). The proposed method could be useful in quality control and standardization of C. grandis 'Tomentosa' raw materials and its products. Results obtained in this study will provide a basis for quality assessment and further study in vivo.