Search

In the past, the PHB gene function was mainly focused on anti-cell proliferation and antitumor effects. But the molecular mechanism of the PHB gene regarding saline and oxidative stresses is unclear. To study the role of AtPHB6 in salt and oxidative stress, AtPHB6 was cloned from A. thaliana. Bioinformatics analysis showed that AtPHB6 was closely related to AtPHB1 and AtPHB2, which are both type II PHB. RT–qPCR results indicated that the AtPHB6 in the leaves and roots of A. thaliana was obviously induced under different stress treatments. AtPHB6-overexpressing plants were larger and more lush than wild-type and mutant plants when placed under stress treatments during seed germination. The root length and fresh weight of AtPHB6 transgenic plants showed the best resistance compared to wild-type plants under different treatments, in contrast, the AtPHB6 mutants had the worst resistance during the seedling stage. AtSOT12 was an interacting protein of AtPHB6, which screened by yeast two-hybrid system. The interaction between the two proteins were further confirmed using in vitro pull-down experiments and in vivo BiFC experiments. Subcellular localization showed both AtPHB6 and AtSOT12 protein expressed in the nucleus and cytoplasm. The H2O2 content in both the transgenic AtPHB6 and AtSOT12 plants were lower than that in the wild type under stresses. Thus, AtPHB6 increased plant resistance to salt stress and interacted with the AtSOT12 protein.

The interpolation of sparse seismic data is important for seismic data processing, especially in the case of marine data acquisition. The conventional interferometric interpolation method interpolates missing data based on seismic interferometry in the time-space domain. However, missing traces and the limited acquisition aperture will reduce the quality and accuracy of virtual shot gathers, thereby affecting the final interpolation results. To improve the conventional method, we have adopted a four-step Radon domain interferometric interpolation method for sparse seismic data. First, we generate synthetic seismic data with a water-layer model and replace missing traces in the input data with synthetic traces. Second, we sort the input shot gathers into common-receiver-point (CRP) gathers and transform the CRP gathers into the Radon domain using a sparse Radon transform method. Third, we perform Radon domain seismic interferometric interpolation by cross-correlating traces with the same ray parameter and summing over all different ray parameters to obtain a virtual shot gather. Fourth, we apply a 1D least-squares matching filter to the virtual shot gather to obtain the interpolation result. Numerical and field examples demonstrate that our method can effectively interpolate sparse seismic data. Compared with the conventional time-space domain interferometric interpolation method, our method can provide more accurate interpolation results. Moreover, it is less sensitive to the depth of the water layer and an inaccurate impedance contrast of the water bottom.

Aster tataricus, a traditional Chinese herb, has been used to treat cough and asthma for many years. Its raw and processed products have different pharmacological effects in clinical applications. To explore the chemical profile differences of components in A. tataricus processed with different methods, metabolomics methods based on ultra-high-performance liquid chromatography coupled with quadrupole time of flight mass spectrometry and gas chromatography-mass spectrometry were developed. Chemometrics strategy was applied to filter and screen the candidate compounds. The accuracy of differential markers was validated by back propagation neural network. The established methods showed that raw A. tataricus, honey-processed A. tataricus, vinegar-processed A. tataricus, and steamed A. tataricus were clearly divided into four groups, suggesting that the components were closely related to the processing methods. A total of 64 nonvolatile and 43 volatile compounds were identified in A. tataricus, and 22 nonvolatile and 12 volatile differential constituents were selected to distinguish the raw and processed A. tataricus. This study demonstrated that the metabolomics methods coupled with chemometrics were a comprehensive strategy to analyze the chemical profile differences and provided a reliable reference for quality evaluation of A. tataricus.

A green, flexible, and effective strategy was proposed to quantify four target compounds (muscone, ethyl palmitate, ethyl oleate, and ethylparaben) from musk by binary eluent based vortex-assisted matrix solid-phase dispersion (MSPD) extraction coupled with GC/MS. Single-factor tests and orthogonal design were employed to optimize the MSPD parameters. In addition, the binary eluent system, methanol, and ethyl acetate 3 : 7 (v/v) were used to extract the target analytes. Finally, C18 was applied as the easily available dispersant and the sample powder was ground for 2 min. Thereafter, the mixture was rapidly extracted with the binary eluents under whirling for 3 min. Eventually, the analysis of the samples was completed within 12 min by GC/MS. All correlation coefficients (r) of four targets were more than 0.9991. The recoveries of all target compounds ranged from 92.8% to 101% while their RSDs were less than 6.94%. There was no significant matrix interference for the analysis. Thus, the combination of vortex-assisted MSPD with GC/MS was considered as a novel, rapid, and environmentally friendly quantitative approach for musk samples.

Asarum is a traditional medicine and has been widely used as remedies for inflammatory diseases, toothache, headache, local anesthesia, and aphthous stomatitis in China, Japan, and Korea. Our previous research found that safrole and methyl eugenol were vital compounds that contribute to distinguish the different species and raw Asarum and its processed products apart. The pharmacokinetics of safrole and methyl eugenol after oral administration of Asarum extract has not been reported yet. In this study, a rapid and simple gas chromatography–mass spectroscopy (GC–MS) method that has a complete run time of only 4.5 min was developed and validated for the simultaneous determination and pharmacokinetic study of safrole and methyl eugenol in rat plasma after administration of Asarum extracts. The chromatographic separation was realized on a DB-17 column ( 30 m × 0.25 mm × 0.25 μ m ). And detection was carried out under selected ion monitoring (SIM) mode. Plasma samples were pretreated by n -hexane. The pharmacokinetic parameters provided by this study will be beneficial for further developments and clinical applications of Asarum.

AtPHB2 was cloned from Arabidopsis thaliana and localized to mitochondria. AtPHB2 expression in A. thaliana increased under different stresses, and promotion of seed germination and plant growth was observed in transgenic plants. In terms of physiological indices, the transgenic plants were more resistant to saline-alkali and oxidative stresses than wild-type and mutant plants. The transgenic plants also had the highest K+/Na+ ratio after 24 h of salt treatment and maintained lower H2O2 contents than wild-type and mutant plants under stress. Taken together, these results reveal that AtPHB2 affects plant tolerance against salt stresses and is involved in ROS decrease mechanisms.

A sensitive and accurate method was developed for the simultaneous determination of twelve components including phenolic acids (gallic acid, protocatechuic aldehyde, protocatechuic acid, and ferulic acid), flavonoids (catechin, epicatechin, rutin, luteolin, luteolin-7-glucoside, and epicatechin gallate), anthraquinones (emodin), and triterpenes (ursolic acid) in Cynomorii herba in different harvest times by liquid chromatography tandem mass spectrometry (LC-MS/MS). The chromatographic separation was achieved on an Eclipse plus C18 (3.0 mm × 50 mm, 1.8 μm) column at 40°C. The mobile phase consisted of acetonitrile and 0.05% formic acid with a gradient elution at a flow rate of 0.4 mL·min −1. Under the optimized conditions, there was good linear relation (r ≥ 0.9944) and satisfactory precision (RSD values less than 5.65%). The recoveries of the twelve components were in the range of 93.5–105%. Moreover, the limits of detection (LOD) ranged from 0.003 to 21 ng mL−1 for the twelve analytes. In conclusion, the validated method was successfully applied to analyze the change regularity of the twelve components of Cynomorii herba in different harvest times. It provides a theoretical basis for choosing the suitable harvesting time of Cynomorii herba.

Background: The ligand-receptor interaction plays an important role in signal transduction required for cellular differentiation, proliferation, and immune response process. The analysis of ligand-receptor interactions is helpful to provide a deeper understanding of cellular proliferation/ differentiation and other cell processes. Methods: The computational technique would be used to promote ligand-receptor interactions research in future proteomics research. In this paper, we propose a novel computational method to predict ligand-receptor interactions from amino acid sequences by a machine learning approach. We extract features from ligand and receptor sequences by Histogram of Oriented Gradient (HOG) and Discrete Cosine Transform (DCT). Then, these features are fed into the Fuzzy C-Means (FCM) clustering algorithm for clustering, and also we get multiple training subsets to generate the same number of sub-classifiers. We choose an optimal sub-classifier for predicting ligand-receptor interactions according to the similarity from one sample to training subsets. Observations: In order to verify the performance, we perform five-fold cross-validation experiments on a ligand-receptor interactions dataset and achieve 80.08% accuracy, 82.98% sensitivity and 80.02% specificity. Then, we test our extracted feature method on two Protein-Protein Interactions (PPIs) datasets, and achieve accuracies of 93.79% and 87.46%, respectively. Conclusion: Our proposed method can be a useful tool for identifying of ligand-receptor interactions. Related data sets and source code are available at https://github.com/guofei-tju/ligand-receptorinteractions. git.

Since the 21st century, energy and environmental issues have become the focus of global attention. Traditional fossil energy resources such as oil and natural gas are drying up. Meanwhile, a large number of harmful gases and other pollutants would be produced in the use of traditional energy, which has a serious damage to the environment and climate. With the rapid increase of energy consumption and the need of environmental protection, the demand for clean and efficient energy is growing. Fuel cell (FC) is considered as one of the most promising energy conversion technologies that can replace conventional fossil fuels, and it is also an important choice in the use of hydrogen, methanol, ethanol and other renewable energy. As a new energy with great potential, FC can reduce the energy crisis and environmental pollution effectively by directly converting the chemical energy of the external input fuel into electrical energy and continuously supplying power to the outside. Ion exchange membrane (IEM) is an important component of FC, which is responsible for transferring ions and forming a complete FC circuit. According to the types of conducting ions, IEM can be divided into proton exchange membrane (PEM) and anion exchange membrane (AEM), which are applied to proton exchange membrane fuel cell (PEMFC) and alkaline anion exchange membrane fuel cell (AAEMFC), respectively. Compared with PEMFC, AAEMFC has the advantages of using non noble metal catalyst, high electrode reaction reactivity, and low fuel purity requirements and so on. However, the commercial AEM with organic cations often has low ionic conductivity and poor alkali and dimensional stability, which seriously hinders its development and application in AAEMFC. Therefore, it is urgent to design and develop new high-performance AEM materials. In order to improve the conductivity, alkali stability and dimensional stability of AEMs, a novel reactive bisphenol monomer with pendant dimethylphenoxybenzene group, 1,1-bis (4-hydroxyphenyl)-1-(4-(3,5-dimethylphenoxy) phenyl) phenyl-2,2,2-trifluoroethane ( 2 ), was synthesized based on molecular design using 4-fluoro-2,2,2-trifluoroacetophenone, 3,5-dimethylphenol and phenol as starting materials by a two-step organic reaction. Then, two series of poly(aryl ether sulfone) anion exchange membranes (2-PAES-QA- xx and 2-PAES-Im- xx ) with comb structure were prepared through aromatic nucleophilic copolycondensation, bromination and quaternization reaction based on monomer 2 . The structures of the active bisphenol monomer, the poly(aryl sulfone) containing methyl groups, the brominated poly(aryl ether sulfone)s and the ionized polymers were characterized systematically by nuclear magnetic resonance spectroscopy, respectively. The water uptake, swelling ratio, ionic conductivity, thermal stability, mechanical properties, and alkaline resistance stability of the prepared 2-PAES-QA- xx and 2-PAES-Im- xx membranes were studied in detail. The relationship between the structure and properties of different types of AEMs is further compared. The obtained AEMs showed good overall performance, and the OH– conductivity of these membranes at 60°C is 35.9–51.6 and 32.6–44.9 mS/cm, respectively. The IEC values of the representative 2-PAES-QA-60 and 2-PAES-Im-60 maintained at 79.7% and 88.5% of the initial values after immersion in 4 mol/L NaOH at 80°C for 336 h, respectively. This work provides a new strategy for the design and preparation of comb-type AEM materials.

Hydrosoluble trehalose lipid (a biosurfactant) was employed for the first time as a green extraction solution to extract the main antioxidant compounds (geniposidic acid, chlorogenic acid, caffeic acid, and rutin) from functional plant tea (Eucommia ulmoides leaves). Single-factor tests and response surface methodology were employed to optimize the extraction conditions for ultrasound-assisted micellar extraction combined with ultra-high-performance liquid chromatography in succession. A Box-Behnken design (three-level, three-factorial) was used to determine the effects of extraction solvent concentration (1-5 mg/mL), extraction solvent volume (5-15 mL), and extraction time (20-40 min) at a uniform ultrasonic power and temperature. In consequence, the best analyte extraction yields could be attained when the trehalose lipid solution concentration was prepared at 3 mg/mL, the trehalose lipid solution volume was 10 mL and the extraction time was set to 35 min. In addition, the recoveries of the antioxidants from Eucommia ulmoides leaves analyzed by this analytical method ranged from 98.2 to 102%. These results indicated that biosurfactant-enhanced ultrasound-assisted micellar extraction coupled with a simple ultra-high-performance liquid chromatography method could be effectively applied in the extraction and analysis of antioxidants from Eucommia ulmoides leaf samples.

In this study, a sensitive and reliable HPLC-MS/MS method was established to quantify tamarixetin in rat plasma. This method was then applied to research on the pharmacokinetic and bioavailability of tamarixetin after intravenous and oral administrationin vivo. The study was performed on CORTECS C18column (4.6 mm × 150 mm, 2.7 μm) using mobile phase composed of methanol-water-formic acid (85 : 15 : 0.1, v/v) at a flow rate of 0.3 mL/min by a tandem mass system with an electrospray ionization (ESI) source in the negative multiple-reaction monitoring (MRM) mode. The calibration curves showed good linearity in the range of 5–4000 ng/mL. The intra- and interday precision of tamarixetin was less than 8.7% and 4.8%, respectively, and accuracy was within ±9.5%. Extraction recovery (91.4–100.0%) and matrix effect (99.4–107.4%) met the guidelines published by regulatory authorities. The oral bioavailability of tamarixetin was 20.3 ± 12.4%.

This investigation was conducted to elucidate whether atractylenolide-I (ATL-1), which is the main component of Atractylodes macrocephala Koidz, can sensitize triple-negative breast cancer (TNBC) cells to paclitaxel and investigate the possible mechanism involved. We discovered that ATL-1 could inhibit tumor cell migration and increase the sensitivity of tumor cells to paclitaxel. ATL-1 downregulated the expression and secretion of CTGF in TNBC cells. Apart from inhibiting TNBC cell migration via CTGF, ATL-1 downregulated the expression of CTGF in fibroblasts and decreased the ability of breast cancer cells to transform fibroblasts into cancer-associated fibroblasts (CAFs), which in turn increased the sensitivity of TNBC cells to paclitaxel. In a mouse model, we found that ATL-1 treatments could enhance the chemotherapeutic effect of paclitaxel on tumors and reduce tumor metastasis to the lungs and liver. Primary cultured fibroblasts derived from inoculated tumors in mice treated with ATL-1 combined with paclitaxel expressed relatively low levels of CAF markers. Collectively, our data indicate that ATL-1 can sensitize TNBC cells to paclitaxel by blocking CTGF expression and fibroblast activation and could be helpful in future research to determine the value of ATL-1 in the clinical setting.

Breast cancer remains the most common malignancy and the leading causality of cancer-associated mortality among women worldwide. With proven efficacy, Oldenlandia diffusa has been extensively applied in breast cancer treatment in Traditional Chinese Medicine (TCM) for thousands of years. However, the bioactive compounds of Oldenlandia diffusa accounting for its anti-breast cancer activity and the underlying biological mechanisms remain to be uncovered. Herein, bioactivity-guided fractionation suggested ursolic acid as the strongest anti-breast cancer compound in Oldenlandia diffusa. Ursolic acid treatment dramatically suppressed the proliferation and promoted mitochondrial-mediated apoptosis in breast cancer cells while brought little cytotoxicities in nonmalignant mammary epithelial cells in vitro. Meanwhile, ursolic acid dramatically impaired both the glycolytic metabolism and mitochondrial respiration function of breast cancer cells. Further investigations demonstrated that ursolic acid may impair the glycolytic metabolism of breast cancer cells by activating Caveolin-1 (Cav-1) signaling, as Cav-1 knockdown could partially abrogate the suppressive effect of ursolic acid on that. Mechanistically, ursolic acid could activate SP1-mediated CAV1 transcription by promoting SP1 expression as well as its binding with CAV1 promoter region. More meaningfully, ursolic acid administration could dramatically suppress the growth and metastasis of breast cancer in both the zebrafish and mouse xenotransplantation models of breast cancer in vivo without any detectable hepatotoxicity, nephrotoxicity or hematotoxicity. This study not only provides preclinical evidence supporting the application of ursolic acid as a promising candidate drug for breast cancer treatment but also sheds novel light on Cav-1 as a druggable target for glycolytic modulation of breast cancer.

This paper presents a new high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method with a rapid analysis of 6 min to determine the concentration of galgravin in rat plasma so as to study its pharmacokinetic features and bioavailability in vivo. Schisandrin was selected as the internal standard (IS). After extracting the analyte from plasma samples with ethyl acetate, methanol-H2O (0.1% formic acid) (85 : 15, v / v ) was used as mobile phase to achieve chromatographic separation on a C18 reversed phase column. The MS detection was performed in positive ion mode using electrospray ionization (ESI) source. This method showed good linearity over the range of 1~500 ng/mL ( R 2 > 0.999 ), and the lower limit of quantitation (LLOQ) was 1.0 ng/mL. The intraday precision and interday precision were both within 8.5%, whereas the accuracies were in the range of -2.6%–6.0%. The average recoveries of galgravin in rat plasma were between 92.3% and 99.3%. Moreover, galgravin was stable throughout storage and processing with all RSDs below 12.1%. After the successful application of this optimized method, the oral bioavailability of galgravin was determined to be 8.5%. This study will be helpful to the future research and development of galgravin.

Background Circular RNAs (circRNAs) crucially regulate tumor progression. In this study, we examined the functional roles and mechanisms of hsa_circ_0003489 in multiple myeloma (MM). Methods Upon altering the expressions of hsa_circ_0003489, miR-874-3p, and/or histone deacetylase 1 (HDAC1) in MM1.R cells and treating them with Bortezomib (BTZ), cell viability was examined by CCK-8 assay, cell proliferation by Ki-67 immunofluorescence, apoptosis by TUNEL staining, flow cytometry and western blot, autophagy by electron microscopy and western blot. The interaction between hsa_circ_0003489 and miR-874-3p as well as that between miR-874-3p and HDAC1 were examined by expressional analysis, dual luciferase reporter assay, and RNA immunoprecipitation. The in vivo impacts of hsa_circ_0003489 on MM growth and sensitivity to BTZ were examined using an MM xenograft mouse model. Results Knocking down hsa_circ_0003489 significantly inhibited the viability, cell proliferation, and the autophagy, while promoted the apoptosis of MM cells in vitro and MM xenograft in vivo. Suppressing hsa_circ_0003489 also further boosted the cytotoxic effects of BTZ in MM cells and reversed its promoting effect on autophagy. Mechanically, hsa_circ_0003489 acted as a sponge of miR-874-3p and positively regulated the expression of miR-874-3p target, HDAC1. MiR-874-3p and HDAC1 essentially mediated the effects of hsa_circ_0003489 on cell viability, proliferation, apoptosis, and autophagy. Conclusion The hsa_circ_0003489/miR-874-3p/HDAC1 axis critically regulates the balance between apoptosis and autophagy. Silencing hsa_circ_0003489 sensitizes MM cells to BTZ by prohibiting autophagy and thus may boost the therapeutic effects of BTZ.

A high-throughput HPLC-MS/MS method was developed and validated for the determination of four antihypertensive drugs including metoprolol tartrate, hydrochlorothiazide, nifedipine, and valsartan in rat plasma. The Sprague-Dawley rats were randomly divided into three groups: A Group: gastric-administration of metoprolol tartrate, hydrochlorothiazide, nifedipine, or valsartan; B Group: a single intravenous injection of SXT, then dosing as the A group; C Group: daily injection of SXT through the tail vein for 8 consecutive days and dosing as the A group on the eighth day. For metoprolol tartrate and valsartan, blood samples were collected before administration and at time points 0.03, 0.08, 0.17, 0.25, 0.5, 1, 2, 4, 6, 8, 10, 12, and 24 h from the fossa orbitalis vein. For hydrochlorothiazide and nifedipine, the time points were 0, 0.08, 0.17, 0.25, 0.5, 0.75, 1, 2, 4, 6, 8, 10, 12, and 24 h. The plasma samples containing different individual antihypertensive drug were mixed and prepared by protein precipitation with methanol. The chromatographic separation was performed on an Agilent Eclipse Plus C18 column (2.1 mm×100 mm, 3.5 μm) using gradient elution with mobile phase consisting of acetonitrile and water (containing 0.1% formic acid). The flow rate was 0.3 mL/min. The detection was accomplished on a tandem mass spectrometer with an electrospray ionization (ESI) source by multiple reaction monitoring (MRM) in both positive and negative modes. The method was successfully applied to a pharmacokinetic interaction study of Shuxuetong injection on the antihypertensive drugs. The results suggested that SXT could increase the total amount of metoprolol tartrate and nifedipine in plasma and showed little influence on the pharmacokinetic behaviors of hydrochlorothiazide and valsartan.

Ultrasmall spherical magnetite nanoparticles (Fe3O4 NPs) coated with one single carbon layer (Fe3O4@1C NPs) were directly synthesized by a simple, fast one-step hydrothermal reaction at low temperature (200 °C). The as-synthesized Fe3O4@1C NPs were easily separated and purified from the resultant mixture, without the need for any additional energy input and chemicals. The as-purified Fe3O4@1C NPs not only displayed typical superparamagnetic behavior but also exhibited quite good long-term stability after being exposed to the open air under the room conditions for more than 2 months. Importantly, the long-term stored Fe3O4@1C NPs exhibited a highly sensitive surface-enhanced Raman spectroscopy (SERS) response toward Rhodamine-B (RdB) molecules with low concentration. The multifunctional and recyclable Fe3O4@1C NPs presented a prospective application for the selective enrichment and sensitive SERS detection of chemical and biomolecular assays.

An in-capillary 2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)-sweeping micellar electrokinetic chromatography-diode array detector (ABTS+-sweeping MEKC-DAD) method was developed and successfully applied to screening and quantifying antioxidative ingredients from natural products. The parameters affecting sweeping and separation were optimized including components of background electrolyte and sample matrix. Comparing with previously reported MEKC, the sensitivity enhancement factors of trace natural antioxidants obtained by this proposed method were from 17 to 167. The limit of detection was as low as 6 ng·mL−1. The results of other validation parameters including linearity, reproducibility, accuracy and recovery were satisfactory. Seven compounds including schizandrin, schisandrol B, schisantherin B, schisantherin A, schisanhenol, deoxyschizandrin, schisandrin B were identified as the main antioxidants in Schisandra chinensis. It was demonstrated that this developed in-capillary ABTS+-sweeping MEKC-DAD is simple, sensitive, reliable and rapid method for screening and quantifying trace antioxidants from natural products.

A novel ionic-liquid-based vortex-simplified matrix solid-phase dispersion method using 2,6-dimethyl-β-cyclodextrin was established by ultra high performance liquid chromatography coupled with a photodiode array detector. 2,6-Dimethyl-β-cyclodextrin was first used as a promising adsorbent in this proposed method for simultaneous determination of eight compounds in Gardeniae fructus. These compounds are terpenoids (geniposidic acid, genipin-1-β-D-gentiobioside, geniposide, 8-o-acetyl shanzhiside methyl ester), crocins (crocin-I, crocin-II), quinic acid derivatives (chlorogenic acid), and flavonoids (isoquercitrin), respectively. Several parameters were investigated in the adsorption and desorption processes to obtain the optimal conditions, including 2,6-dimethyl-β-cyclodextrin as sorbent, 0.5 mL 100 mM 1-dodecyl-3-methylimidazolium hydrogen sulfate as the extraction solvent, 2:1 of sample/sorbent ratio, grinding for 2 min and vortexing for 60 s. The recoveries of the eight compounds ranged from 96.6 to 100% (3.50%). The limits of detection and quantification were in the range of 0.02-0.30 and 0.06-1.25 μg/mL, respectively. Meanwhile, a good linearity was attained with r values (0.9997). The established method showed higher extraction efficiency and less reagent consumption than traditional matrix solid phase dispersion and ultrasonic-assisted extraction. Hence, it could be applied for sample preparation and analysis of natural products.

An effective ultrasonic-assisted extraction method for the separation of phenolic acids and flavonoids in Cuscuta chinensis Lam. was conducted by combining uniform design (UD) coupled with response surface methodology (RSM) and orthogonal design (OD) experiment. A sensitive and selective high-performance liquid chromatography-electrospray ionization tandem triple quadrupole mass spectrometry (HPLC-ESI-MS/MS) method was applied to quantify the sixteen active ingredients (chlorogenic acid, cryptochlorogenic acid, neochlorogenic acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, caffeic acid, hyperin, isoquercitrin, quercetin, campherol, p-coumaric acid, isorhamnetin, rutin, astragalin, and apigenin). The extraction method was optimized with respect to concentration of extraction solvent, extraction time, and ratio of liquid to solid as a consequence of getting a high sensitive and feasible method for simultaneous determination of contents of multiple components and evaluation of quality control of Cuscuta chinensis Lam. from different origins. It was also considered useful and valuable in the further study for quality control of Cuscuta chinensis Lam.

Baijiu is an important fermented product in China. A yeast named YX3307 that is capable of producing a large amount of ethyl caproate (EC) was isolated from Daqu, a crude fermentation starter for Baijiu. This yeast was identified as Clavispora lusitaniae on the basis of its morphological properties, physiological and biochemical characteristics, and 26S rDNA sequence. Single-factor experiments were conducted to obtain the optimum fermentation conditions for EC production by YX3307. The highest EC yield (62.0 mg/L) from YX3307 was obtained with the following culture conditions: inoculum size 7.5% (v/v), seed cell age 30 h, sorghum hydrolysate medium (SHM) with a sugar content of 10 Brix and an initial pH of 6.0; incubation at 28°C with shaking at 180 rpm for 32 h; addition of 10% (v/v) anhydrous ethanol and 0.04% (v/v) caproic acid at 32 and 40 h, respectively, static culture at 20°C until 72 h. YX3307 synthesized more EC than ethyl acetate, ethyl lactate, ethyl butyrate, and ethyl octanoate. An intracellular enzyme or cell membrane enzyme was responsible for EC synthesis. YX3307 can produce many flavor compounds that are important for high-quality Baijiu. Thus, it has potential applications in improving the flavor and quality of Baijiu.

This study aimed to extract polysaccharides from citron and analyze their structures and potential bioactivities. Two novel polysaccharides CM-1 and CM-2 were purified from citron by DEAE-Sepharose Fast Flow and Sephadex G-100 column chromatography. Monosaccharide composition, linkage and NMR data were used to infer their sugar chains composition. The anti-breast cancer cells and immunoregulatory activities of CM-1 and CM-2 were investigated. Results indicated that CM-1 (Mw = 21,520 Da), composed of arabinose, xylose, mannose and glucose in a molar ratio of 10.78:11.53:1.00:1.70, was arabinoxylan (AX) with (1 → 4)-linked β- d -Xylp skeleton monosubstituted with α- l -Araf units at O-3 position. While CM-2 (Mw = 22,303 Da), composed of arabinose, mannose, glucose and galactose in a molar ratio of 25.46:1.45:1.00:6.57, was galactoarabinan (GA) with (1 → 5)-linked α- l -Araf backbone substituted by β- d -Galp units at O-2 and/or O-3 positions. Both polysaccharides exhibited potential inhibiting cancer and immunostimulatory activities in vitro, especially CM-1. These results provide a basis for further research on citron polysaccharides.

Isoflavones (daidzein, daidzin, genistein and genistin) were main chemical components and usually selected as markers for quality control of Traditional Chinese Medicine semen sojae praeparatum (SSP). High throughput vibration and vortex-assisted matrix solid phase dispersion and high performance liquid chromatography with diode array detection were developed to simultaneously extract and quantify four isoflavones in SSP. Some parameters influencing extraction efficiency of isoflavones by vortex-assisted matrix solid phase dispersion such as sorbent type, ratio of sample to sorbent, crushing time, vibration frequency, methanol concentration, eluting solvent volume and vortex time were optimized. It was found that the best extraction yields of four isoflavones were obtained when the sample (20 mg) and SBA-3 (40 mg) was crushed by ball mill machine for 2 min at vibration frequency of 800 times per minute. Methanol/water (1.5 ml, 8:2, v/v) solution was dropped into the treated sample and vortexed for 3 min. The recoveries of the four isoflavones ranged from 86.1 to 94.8% and all relative standard deviations were less than 5%. A good linearity (r > 0.9994) was achieved within the range 0.5–125 μg/ml. It was concluded that the high throughput vibration and vortex-assisted matrix solid-phase dispersion coupled with high performance liquid chromatography was user-friendly extraction and quantification method of multiple isoflavones for quality evaluation of SSP.

The emergence of antibiotic tolerance was a tricky problem in the treatment of chronic Pseudomonas aeruginosa-infected cystic fibrosis and burn victims. The quorum sensing (QS) inhibitor may serve as a new tactic for the bacterial resistance by inhibiting the biofilm formation and the production of virulence factors. This study explored the potential of luteolin as a QS inhibitor against P. aeruginosa and the molecular mechanism involved.Crystal violet staining, CLSM observation, and SEM analysis were carried out to assess the effect of luteolin on biofilm formation. The motility assays and the production of virulence factors were determined to evaluate the QS-inhibitory activity of luteolin. Acyl-homoserine lactone, RT-PCR, and molecular docking assays were conducted to explain its anti-QS mechanisms.The biofilm formation, the production of virulence factors, and the motility of P. aeruginosa could be efficiently inhibited by luteolin. Luteolin could also attenuate the accumulation of the QS-signaling molecules N-(3-oxododecanoyl)-L-homoserine lactone (OdDHL) and N-butanoyl-L-homoserine lactone (BHL) (P 0.01) and downregulate the transcription levels of QS genes (lasR, lasI, rhlR, and rhlI) (P 0.01). Molecular docking analysis indicated that luteolin had a greater docking affinity with LasR regulator protein compared with OdDHL.This study is important as it reports the molecular mechanisms involved in the anti-biofilm formation activity of luteolin against P. aeruginosa. This study also indicated that luteolin could be helpful when used for the treatment of clinical drug-resistant infections of P. aeruginosa.

Glypican-3 (GPC3) is a highly specific diagnostic marker for hepatocellular carcinoma (HCC) diagnosis and a potential target in HCC therapy. Nanobodies (Nbs) are promising targeting molecules due to their high specificity and strong affinities to antigens, high stability, deep tissue penetration, and low immunogenicity. In this study, we isolated Nbs against GPC3 marker protein from a synthetic Nb library by phage display. To characterize these Nbs, we performed enzyme-linked immunosorbent assay, immunoprecipitation assay, and immunofluorescent assay to demonstrate that four (G8, G10, G11, and G64) of them bound specifically to recombinant as well as endogenous GPC3, and epitope mapping showed they all bound to N-terminal subunit of GPC3. Furthermore, we found that G64 exhibited high protein stability and GPC3 binding activity in serum at 37℃ for at least 96 h, and G64 did not affect the proliferation of HEK293T cells and HCC cell line HepG2. Our study provides four anti-GPC3 Nbs as promising targeting molecules for HCC diagnostic and therapeutic drugs.

The strategy of decreasing atherosclerotic cardiovascular disorder is imperative for reducing premature death and improving quality of life in patients with diabetes mellitus. The aim of this study was to investigate whether the natural flavone acacetin could protect against endothelial injury induced by high glucose and attenuate diabetes-accelerated atherosclerosis in streptozotocin-(STZ) induced diabetic ApoE−/− mice model. It was found that in human umbilical vein endothelial cells (HUVECs) cultured with normal 5.5 mM or high 33 mM glucose, acacetin (0.3–3 μM) exerted strong cytoprotective effects by reversing high glucose-induced viability reduction and reducing apoptosis and excess production of intracellular reactive oxygen species (ROS) and malondialdehyde in a concentration-dependent manner. Acacetin countered high glucose-induced depolarization of mitochondrial membrane potential and reduction of ATP product and mitoBcl-2/mitoBax ratio. Silencing Sirt3 abolished the beneficial effects of acacetin. Further analysis revealed that these effects of acacetin rely on Sirt1 activation by increasing NAD+ followed by increasing Sirt3, pAMPK and PGC-1α. In STZ-diabetic mice, acacetin significantly upregulated the decreased signaling molecules (i.e. SOD, Bcl-2, PGC-1α, pAMPK, Sirt3 and Sirt1) in aorta tissue and attenuated atherosclerosis. These results indicate that vascular endothelial protection of acacetin by activating Sirt1/Sirt3/AMPK signals is likely involved in alleviating diabetes-accelerated atherosclerosis by preserving mitochondrial function, which suggests that acacetin may be a drug candidate for treating cardiovascular disorder in patients with diabetes.

Chronic stable angina (CSA) is caused by coronary atherosclerosis. The gut microbiota (GM) and their metabolite trimethylamine-N-oxide (TMAO) levels are associated with atherosclerosis. Danlou tablet (DLT) combined with Salvia miltiorrhiza ligustrazine (SML) injection has been used to treat CSA. This study aims to investigate how DLT combined with SML (DLT-SML) regulates serum lipids, inflammatory cytokines, GM community, and microbial metabolite in patients with CSA. In this study, 30 patients with CSA were enrolled in the DLT-SML group, and 10 healthy volunteers were included in the healthy control group. The patients in the DLT-SML group were subdivided as the normal total cholesterol (TC) group and high-TC group according to their serum TC level before treatment. Blood samples were collected to investigate the (1) lipid content, including triglyceride (TG), TC, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol, (2) fasting blood glucose (Glu), (3) inflammatory cytokines, including interleukin-1 beta (IL-1β), interleukin-6 (IL-6), interleukin-10 (IL-10), and tumor necrosis factor-α (TNF-α), and (4) gut-derived metabolite, including lipopolysaccharides and TMAO level. GM composition was analyzed by sequencing 16S rRNA of fecal samples. Results showed that DLT-SML significantly decreased serum TG, TC, low-density lipoprotein cholesterol, IL-1β, TNF-α, and TMAO levels of patients with CSA. DLT-SML increased the abundance of Firmicutes and decreased Proteobacteria, which were significantly lower or higher in patients with CSA, respectively, compared with the healthy control group. In particular, DLT-SML increased the microbial diversity and decreased Firmicutes/Bacteroidetes ratio of patients with high-TC. The abundance of Sarcina, Anaerostipes, Streptococcus, Weissella, and Erysipelatoclostridium was decreased, whereas Romboutsia, Faecalibacterium, and Subdoligranulum were increased by DLT-SML treatment in patients with CSA. These findings indicated that DLT-SML improved patients with CSA by ameliorating dyslipidemia profile, decreasing the circulating inflammatory cytokines, and regulating the GM composition and their metabolites.

Fluorescent carbon dots (CDs) were directly synthesized by hydrothermal treatment of medicinal mulberry leaves (Morus alba L.) residues without the need for any chemical reagent. The CDs were collected successively through filtration, centrifugation, and dialysis from the resultant solution. The as-purified CDs were almost non-cytotoxic and successfully used for the intracellular fluorescence imaging of human hepatocellular carcinoma cells (HepG2). Due to the presence of plentiful surface groups, the CDs were further used as drug carriers for the loading of Lycorine, an anti-cancer alkaloidal extracted from bulbs of Lycoris radiata L. Importantly, the Lycorine-loaded CDs were successfully applied for simultaneous intracellular imaging and drug delivery for the better anti-cancer cell activity. The present work both provides a strategy for the reuse of medicinal herb residues for the preparation of CDs and promotes a potential application of CDs in biomedicine and bioimaging.

BackgroundThe strategy of decreasing atherosclerotic cardiovascular disorder is imperative to reduce premature death and improve quality of life in patients with diabetes mellitus. The present study was designed to investigate whether the natural flavone acacetin could improve diabetes- accelerated atherosclerotic lesions.MethodsDiabetic model was established in 7-week-old ApoE−/− mice by intraperitoneal injection of STZ (daily 50 mg/kg) for 5 days. Animals of control, control with acacetin treatment, STZ-diabetes, STZ-diabetes with acacetin treatment received acacetin prodrug subcutaneously (20 mg/kg, b.i.d.) or equivolume saline for 12 weeks, and the elasticity of carotid artery and the ability of vascular wall movement were determined with ultrasound and magnetic resonance imaging. Human umbilical vein endothelial cells (HUVECs) were cultured with medium containing 5.5 mM or 33 mM glucose and treated with acacetin or vehicle. Changes of related aortic lesions and signaling molecules were determined by biochemical and molecular approaches in animals and cultured HUVECs.ResultsIt was found that acacetin significantly suppressed atherosclerotic lesions and neointima hyperplasia, improved the elasticity of carotid artery and the ability of vascular wall movement without reducing blood glucose level and reversed the impaired signaling molecules (i.e. SOD, Bcl2, PGC-1α, pAMPK, Sirt3 and Sirt1) in artery tissues in diabetic mice. In cultured HUVECs, high glucose-induced cell viability reduction, ROS over-production, decrease of anti-oxidation, increase of apoptosis, and impairment of mitochondrial function were countered by acacetin (0.3-3 µM) in a concentration-dependent manner. Moreover, acacetin relies on Sirt1 activation by increasing NAMPT and NAD+ followed by Sirt3, pAMPK and PGC-1α activation. Silencing Sirt1 abolished acacetin-induced activation of Sirt3, pAMPK, and PGC-1α.ConclusionsThese results indicate that Sirt1-mediated activation of pAMPK/Sirt3 signals is involved in protective effects of acacetin against atherosclerosis in diabetes by preserving mitochondrial function via reducing mitochondrial apoptosis and ROS production and enhancing its biogenesis, which suggests that acacetin may be a drug candidate for reducing atherosclerotic cardiovascular disorder in patients with diabetes.

Ethnopharmacological relevance Coptis chinensis Franch (CCF), also known as Huang Lian in China, is a traditional Chinese medicine that commonly used for more than 2000 years. Clinically, CCF often used as anti-inflammatory, immune regulation and other effects. It has been reported that the decoction containing CCF can be used for the treatment of benign prostatic hyperplasia (BPH) or lower urinary tract symptoms (LUTS). Aim of the study: This research aims to investigate the effect of CCF on inhibition of BPH development in vivo and in vitro, and further identify the active compound (s) and the possible mechanism involved in BPH-related bladder dysfunction. Materials and methods Oestrodial/testosterone-induced BPH rat model was established as the in vivo model. The prostate index (PI) was calculated, the pathogenesis was analyzed and the micturition parameters were determined in the shamed-operated, BPH model and BPH + CCF groups after 4-week administration. The tension in detrusor strips was then assessed upon KCl or ACh stimulation with or without incubation of CCF or active compounds. To further investigate the signaling involved, rat detrusor cells were cultured as the in vitro models, the instantaneous calcium influx was measured and the ROCK-1 expression was detected. Results Increased PI value and the aggravated prostatic pathology were observed with voiding dysfunction in BPH rats, which were significantly blocked by oral CCF taken. ACh or KCl-induced contractile responses in detrusor strips were significantly inhibited and the micturition parameters were improved when incubation with CCF or its active compounds such as berberine. Both CCF and berberine suppressed the cellular calcium influx and ROCK-1 expression upon ACh stimulation, demonstrating that berberine was one of the active compounds that contributed to CCF-improved micturition symptoms and function. Conclusions Taken together, our findings give evidence that CCF and its active compound berberine inhibited BPH and bladder dysfunction via Ca2+ and ROCK signaling, supporting their clinical use for BPH and BPH-related LUTS treatment.

The controllable synthesis of nanosized Fe3O4 (10–20 nm) encapsulated in different numbers of graphene layers (1–5 layers) (Fe3O4@DGL NPs) was realized through a facile and green hydrothermal reaction at a temperature as low as 200 °C. The competitive reduction–oxidation between reducing ethylene glycol (EG) and oxidizing H2O under hydrothermal conditions resulted in the emergence of a magnetic Fe3O4 core. Then, the pyrolytic reaction of the polyvinyl alcohol (PVA) molecules attached to the surface of the Fe3O4 core with different surface densities led to the formation of graphene with a controlled number of layers. These Fe3O4@DGL NPs exhibited fast adsorption and sensitive SERS detection for rhodamine B (RhB). A physical and mathematical model was proposed for the estimation of the enhancement factor (EF) by combining the adsorption efficiency and SERS of RhB. This approach and model are applicable for the adsorption, sensitive SERS detection and determination of SERS EF when using functional magnetic nanoparticles as the adsorbent. The Fe3O4@1G NPs were also used as a novel nano-adsorbent for the fast removal of Escherichia coli (E. coli) from an aqueous solution. The Fe3O4@1G NPs regenerated after 3 cycles also showed high efficiency in the adsorption and separation of RhB and E. coli.

Ethnopharmacological relevance San-Ye-Tang-Zhi-Qing formula (SYTZQ) is an effective prescription for the treatment of pre-diabetes disorders of glycolipid metabolism in type 2 diabetes mellitus (T2DM). It consists of five Chinese herbs including Mori Folium, Nelumbinis Folium, Crataegi Folium, Salviae Miltiorrhizae Radix et Rhizoma and Paeoniae Radix Rubra. Aim of the study This study was aimed to reveal the pharmacological mechanism of pharmacokinetic target components of SYTZQ for the treatment of T2DM. Materials and methods A rapid, precise and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed to quantify simultaneously nuciferin, vitexin-4″-O-glucoside, vitexin-2″-O-rhamnoside, paeoniflorin and rosmarinic acid in rat plasma after oral administration of SYTZQ. The network pharmacology was used to analyze the effect of the compounds absorbed into the blood of SYTZQ on T2DM. The effects of paeoniflorin, nuciferine and rosmarinic acid on adipogenic differentiation were validated in vitro experiments. Results The separation was performed on an ACQUITY UHPLC HSS T3 column (2.1 mm × 100 mm, 1.7 μm) using acetonitrile and 0.1% (v/v) formic acid in water as the mobile phase in gradient elution. The calibration curves of five analytes showed good linearity (r ≥ 0.9991) with the lower limits of quantification (LLOQ) between 0.3 and 5.0 ng/mL. The recoveries and matrix effects of five analytes ranged from 81.1% to 113%. The RSDs of inter-day and intra-day precision were all within 13.7%. The validated method was successfully applied to the pharmacokinetic study of five ingredients after oral administration of SYTZQ to rat. 39 major targets and 22 candidate pathways of five compounds absorbed into the blood of rats after administration of SYTZQ were identified and successfully constructed a compound-target-disease-pathway network. It was confirmed that paeniforin, nuciferine and rosmarinic acid could decrease the adipogenicity differentiation in vitro experiments. Conclusions The pharmacokinetic parameters indicated that the five components (nuciferin, vitexin-4″-O-glucoside, vitexin-2″-O-rhamnoside, paeoniflorin and rosmarinic acid) were absorbed and eliminated quickly in vivo. These five absorbed components were associated with 22 pathways, including insulin resistance, regulation of lipolysis in adipocytes, PI3k/AKT-, TNF-, cAMP- and cGMP-PKG-signaling pathway. Paeoniflorin, nuciferine and rosmarinic acid have the effect of inhibiting adipocyte differentiation. This study could provide more reference for quality control, and provide a firm basis for evaluating the clinical efficiency of SYTZQ.

Neuraminidase plays an essential role in the spread of influenza viruses via cleaving sialic acids from the host cell receptors and virions. Neuraminidase has been regarded as an essential target for prevention and treatment of influenza infection. The one-step high-performance liquid chromatography-fraction collector (HPLC-FC) was selected to prepare fractions from Reduning (RDN) injection, while ultra-high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UHPLC/Q-TOF-MS) was used to identify fractions depending on their retention time and molecular ion. As a result, 75 fractions were prepared and 28 fractions out of them exhibited NA inhibitory effects with the dose-effect relationship. Exploring it further, six components including neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, and isochlorogenic acid C were the main components that accounted for almost 80% of inhibitory activity of RDN injection. Accordingly, these results demonstrated that this strategy could not only rapidly identify but also accurately screen active components from traditional Chinese medicine.

The extract of Gardeniae Fructus (GF) with different processing methods processed the different medicinal properties and efficacy. Crude GF (CGF) could be processed into stir-frying GF (SGF), gancao mix-frying GF (GCGF), and ginger mix-frying GF (GIGF) in practice. An LC-MS/MS method was established for simultaneous quantification of geniposidic acid, geniposide, genipin-1-β-gentiobioside, genipin, and crocetin in the rat plasma. The LLOQs for determination of all five components were 10 ng/mL. The accuracies of intraday and interday were in the range of 91%–105%. The recoveries of 5 analytes ranged from 81.0% to 114% with RSD less than 14%. The results showed that the AUCs (area under the plasma concentration-time curve) and Cmax (maximum plasma concentration) of geniposidic acid, genipin-1-β-gentiobioside, and geniposide after oral administration of the CGF extract were apparently higher than those after oral administration of other processed extracts. Cmax of geniposide in plasma after administration of GIGF significantly decreased (p<0.01). Genipin was not detected in rat plasma after administration of the GIGF extract, but it can be detected in plasma after administration of CGF, SGF, and GCGF extract. Furthermore, crocin I and crocin II were not detected in plasma samples. Crocetin had higher concentration in rat plasma versus lower contents in extract. It was demonstrated that the different processing methods might influence the pharmacokinetics of geniposidic acid, genipin-1-β-gentiobioside, geniposide, genipin, and crocetin.

Veratrum nigrum L. is a well-known traditional Chinese medicine with a lot of pharmacological activities including antihypertensive, anticancer, and antifungal effects. In the current experiment, a rapid and sensitive UPLC-MS/MS method that takes only 7 min run time has been established and validated for simultaneous determination of eight bioactive compounds including cyclopamine, jervine, veratramine, polydatin, quercetin, apigenin, resveratrol, and veratrosine in rat plasma. The chromatographic separation of analytes and internal standard was performed on a Phenyl-Hexyl column ( 2.1 × 100 mm , 1.7 μm) with the mobile phase consisting of water (0.1% formic acid) and acetonitrile at a flow rate of 0.3 mL/min. An electrospray ionization (ESI) source was used to detect the samples in both positive and negative ion modes. The intra- and interday precisions of the compounds were less than 9.5% and the accuracy ranged from -10.8% to 10.4%. The extraction recoveries of the compounds were in the range of 85.1 ± 1.5 % to 102.6 ± 8.0 % , and the matrix effect ranged from 91.2 ± 4.5 % to 113.8 ± 1.5 % . According to the results of the stability test, the eight compounds have good stability under various conditions and the relative standard deviation (RSD) less than 13.2%. The pharmacokinetic parameters of the eight compounds in rat plasma after oral administration of Veratrum nigrum L. extract were successfully determined by the established UPLC-MS/MS method.

Red ginseng is often combined with Aconiti Lateralis Radix Praeparata to reduce alkaloids-related toxicity of the latter. Such herb-pairing also results in better therapeutic effect in heart failure, as compared to the singular use of either herb. The purpose of this study was to investigate the effect of Aconiti Lateralis Radix Praeparata and its combination with red ginseng on the activities of CYP450 enzymes in rats. A sensitive and reliable HPLC-MS/MS method was established and validated for the simultaneous determination of eight probe drugs, phenacetin (CYP1A2), tolbutamide (CYP2C9), omeprazole (CYP2C19), dextromethorphan (CYP2D6), dapsone (CYP3A4), 7-hydroxycoumarin (CYP2A6), bupropion (CYP2B6), and amodiaquine (CYP2C8), in rat plasma using diazepam as internal standard (IS). The chromatographic separation was performed on a Waters XBridge™ C18 column (2.1 mm × 100 mm, 3.5 μm) using a gradient elution with the mobile phase consisting of acetonitrile and water (containing 0.1% formic acid) at a flow rate of 0.3 mL/min. The method was successfully applied in evaluating the effect of Aconiti Lateralis Radix Praeparata and red ginseng on the activities of CYP450 enzymes. The pharmacokinetic results of the eight probe drugs suggested that Aconiti Lateralis Radix Praeparata may inhibit the activity of CYP2A6, CYP2C19, CYP2B6, CYP1A2, CYP3A4, and CYP2C9 enzymes in rats. Comparison between the two groups, Aconiti Lateralis Radix Praeparata combined with red ginseng and Aconiti Lateralis Radix Praeparata, indicated that red ginseng may inhibit the activity of CYP2D6 and CYP2B6 enzymes while inducing the activity of CYP1A2, CYP3A4, and CYP2C9 enzymes.

Asarum (Aristolochiaceae) is one of the common herbs used to relieve exterior syndromes. Some volatile components of Asarum which have toxic effect may cause adverse reactions such as headache, general tension, unconsciousness, and respiratory paralysis. Therefore, Asarum is normally processed to reduce such toxicity and adverse effects. The bioactive ingredients contained in different Asarum herbs vary significantly; this variation may be attributed to their differences in species, origins, or processing methods. In this study, 16 batches of Asarum herbs were collected, and their species were identified using DNA barcoding, which is a method for distinguishing plant species, coupled with microscopy. A gas chromatography-mass spectrometry (GC-MS) method for simultaneous determination of 10 compounds was established to evaluate the contents of raw and processed Asarum herbs. Multivariate analysis was then applied to compare different batches of herbs based on the GC-MS data. DNA barcoding identified the herbs as being derived from four sources, and herbs from different origins showed different microscopic features. The results demonstrated that most of the samples were clearly clustered into distinct groups that corresponded to species types. All raw and processed samples were classified by partial least squares discriminant analysis (PLS-DA) based on the 10 analyzed compounds. The findings suggested that safrole and methyleugenol with a variable importance in the project (VIP) > 1 are unique compounds that can be used to differentiate between Asarum species. Safrole, methyleugenol, and 2,6,6-trimethylcyclohepta-2,4-dien-1-one were identified as significant constituents, the presence of which can be used to differentiate between raw and processed Asarum samples. These results indicate that species and processing methods show important effects on the composition of Asarum herbs.

A rapid and reliable UPLC-MS/MS method was developed and validated for the simultaneous quantification of thirteen bioactive compounds (luteolin, cynaroside, luteolin 7-O-glucuronide, isochlorogenic acid C, chlorogenic acid, cryptochlorogenic acid, apigenin, apigenin 7-glucoside, acacetin, hyperoside, isoquercitrin, tilianin, and hesperidin) in rat plasma. The compounds were separated on an ACQUITY UPLC BEH C18 column (2.1×100 mm, 1.7 μm) with a gradient mobile phase system of acetonitrile and 0.1% (v/v) formic acid aqueous solution at a flow rate of 0.3 mL/min. All compounds were quantitated using Agilent Jet Stream electrospray ionization (AJS ESI) in a negative ion mode. The lower limit of quantification (LLOQ) for all compounds was below 5 ng/mL. The intra- and interday accuracy ranged from -13.0% to 14.0%, and precisions were less than 12.2%. The extraction recoveries of the compounds were in the range of 56.9% to 95.0%, and the matrix effect ranged between 71.6% and 109.3%. Stability studies proved that the thirteen compounds were stable under tested conditions, with a relative standard deviation (RSD) of less than 11.4%. This developed method was successfully applied to the pharmacokinetic study of the 13 bioactive compounds after oral administration of Flos Chrysanthemi extract in rat by UPLC-MS/MS. Pharmacokinetic parameters of 8 out of the 13 compounds investigated are presented in this paper.

Photo-Fenton process is an efficient way to treat the organic pollutants in wastewater. However, the efficiency is limited by serious leaching of iron ions, separation of spent catalyst from reaction system for facile recycling, acidic reaction environment (pH＜4). Herein, a novel double perovskite photocatalyst La2AlFeO6 with high surface area was developed to counter these issues. By altering complexing agents, iron ion content discrepancy and B’-O-B” varies in La2AlFeO6 led to a high surface Fe3+ ratio and an excellent magnetic property. >98.4% of phenol could be degraded in a wide pH range over La2AlFeO6 because of the high surface Fe3+ ratio and promotion of generated ·OH, ·O2– and 1O2. The excellent magnetic property would contribute to separate catalyst from water, and led the used La2AlFeO6 to be reused after simple deionized water rinsing and air drying without any further treatment or calcination.

In order to enhance the properties of polymer anion-exchange membranes (AEMs), a class of side-chain-type poly (arylene ether sulfone)s containing tri-imidazole cations in pendant phenoxyphenyl spacer (i.e., Im-PES-xx) were designed and prepared by polymer grafting reaction. Based on these imidazole functionalized polymers, a series of new AEMs were successfully prepared by their solution coating and ion exchange. The obtained membranes exhibited good alkaline stability with the retention of ion-exchange capacity in the range of 78.5%–83.3% after immersion in 4 M NaOH solution at 80 °C for 336 h. The membranes also had high hydroxide conductivity with the values of 34.1–70.5 mS cm−1 at 80 °C. Meanwhile, these membranes possessed good dimensional stability, and their swelling ratio was in the range of 2.5%–7.0% at 80 °C. The incorporation of the dense and local imidazole cations in the side chains for AEMs plays an important role for their properties.

Both magnetic Fe3O4 nanoparticles (Fe3O4 NPs) and luminescent carbon dots (CDs) were simultaneously synthesized via a hydrothermal route at temperature as low as 200 °C. The synthesized magnetic Fe3O4 NPs and luminescent CDs were readily separated and purified from the reaction mixture, without the need for additional energy input and chemicals. The magnetic properties of purified Fe3O4 and photoluminescence (PL) behaviors of purified CDs were investigated using Squid-VSM magnetic property measurement system and fluorescence spectrophotometer, respectively. The size of Fe3O4 NPs and the luminescent properties of CDs were tunable by changing the reaction time or ratio between starting materials within a certain range. Fe3O4 NPs exhibited high saturation magnetization in the range of 36.59–68.22 emu/g and their magnetic behavior significantly depend on the size and structure of Fe3O4 NPs. This work was expected to provide a kind of green and high efficiency simultaneously synthesis of size-controllable Fe3O4 NPs and photoluminescence tunable CDs.

A simple and green sodium dodecyl sulfate-synergistic microwave-assisted extraction method was developed to extract and determine the iridoids, phenylpropanoids, and lignans in Eucommiae Cortex followed by ultra-high-performance liquid chromatography with photodiode array detection. The biodegradable solution (sodium dodecyl sulfate) was used as a promising alternative to organic solvents. The response surface methodology provided the optimum extraction conditions (2 mg/mL sodium dodecyl sulfate, 1100 W microwave power, and 6 min extraction time). The recoveries of three types of components ranged from 95.0 to 105% (RSDs < 5%). The intra- and inter-day precision and accuracy were less than 3.40% and within the range of 97.1-105%, respectively. Compared with other extraction methods, this newly established method was more efficient and environmental friendly. The results demonstrated that sodium dodecyl sulfate-synergistic microwave-assisted extraction followed by ultra-high-performance liquid chromatography with photodiode array method was applicable for the simultaneous extraction and determination of these three types of compounds for quality evaluation of Eucommiae Cortex.

Advanced non-small cell lung cancer (NSCLC) leads to a high death rate in patients and is a major threat to human health. NSCLC induces an immune suppressive microenvironment and escapes from immune surveillance in vivo. At present, the molecular mechanisms of NSCLC immunopathogenesis and the immune suppressive microenvironment induced by NSCLC have not been fully elucidated. Here, we focus on the effect of NSCLC cells on the development and differentiation of human CD1c+ conventional dendritic cell (DC) subsets mediated by CD205 and CD103. The peripheral blood mononuclear cells (PBMCs) were isolated from NSCLC patients and healthy donors. DCs were induced and cocultured with primary NSCLC cells or tumor cell line H1299. DCs without incubation with tumor cells are control. The protein expression of costimulatory molecules such as CD80 and CD86, HLA-DR, pro-/anti-inflammatory cytokines such as IL-10 and IL-12, and CD205 and CD103 on CD1c+ DCs was detected by flow cytometry. Our data revealed two new subpopulations of human CD1c+ DCs (CD1c+CD205+CD103+ and CD1c+CD205+CD103- DC) in healthy donors and NSCLC patients. NSCLC cells modulate the development of the CD1c+CD205+CD103+ DC and CD1c+CD205+CD103- DC subpopulations in vitro and ex vivo. NSCLC cells also suppress the expression of signal molecules such as CD40, CD80, CD86, and HLA-DR on CD1c+ DCs. In addition, the production of pro-inflammatory cytokines, including IL-12 and IL-23, is downregulated by NSCLC cells; however, the secretion of anti-inflammatory cytokines, such as IL-10 and IL-27, by CD1c+ DCs is upregulated by NSCLC cells. Our results suggest that NSCLC cells may induce immune tolerogenic DCs, which block DC-mediated anti-tumor immunity in NSCLC patients. Our data may be helpful in revealing new cellular mechanisms related to the induction of tolerogenic CD1c+ DCs by NSCLCs and the development of an immune suppressive microenvironment that causes tumor cells to escape immune surveillance. Our results indicate a potential role for CD1c+ DC subsets mediated by CD205 and CD103 in DC-mediated immunotherapy to target NSCLC in the future.

Columbianetin-β- d -glucopyranoside (CBG) and its metabolite columbianetin (CBN) are the bioactive constituents of Angelicae pubescentis radix (APR). They exhibit the anti-platelet aggregation, anti-inflammatory and analgesic properties. The absorption, distribution, metabolism and excretion (ADME) of CBG has not been reported to date. Both high-performance liquid chromatography with fluorescence detection and ultra high-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry methods were developed and validated for the study of ADME of CBG. It was found that CBG could be catabolized into its active metabolite CBN in vivo. The absolute bioavailability of columbianetin-β- d -glucopyranoside was 5.63 ± 4.42%. The other co-existing constituents from the APR ethanol extract could enhance the absorption of CBG. CBG and CBN were rapidly and broadly distributed in the stomach, ovary, kidney, liver, spleen, lung, muscles, heart and brain. Higher levels of accumulation of CBG and CBN were detected in the ovary and kidney tissues. Eight metabolites of CBG were tentatively identified in blood, urine, bile and faeces of rats after oral administration of pure CBG. It was also found that CBG and CBN were mainly excreted through the faecal route. It can be concluded that the validated methods were successfully applied for absorption, distribution, metabolism and excretion study of CBG.

A novel method of on-line 2,2′-Azinobis-(3-ethylbenzthiazoline-6-sulphonate)-Capillary Electrophoresis-Diode Array Detector (on-line ABTS+-CE-DAD) was developed to screen the major antioxidants from complex herbal medicines. ABTS+, one of well-known oxygen free radicals was firstly integrated into the capillary. For simultaneously detecting and separating ABTS+ and chemical components of herb medicines, some conditions were optimized. The on-line ABTS+-CE-DAD method has successfully been used to screen the main antioxidants from Shuxuening injection (SI), an herbal medicines injection. Under the optimum conditions, nine ingredients of SI including clitorin, rutin, isoquercitrin, Quercetin-3-O-D-glucosyl]-(1-2)-L-rhamnoside, kaempferol-3-O-rutinoside, kaempferol-7-O-β-D-glucopyranoside, apigenin-7-O-Glucoside, quercetin-3-O-[2-O-(6-O-p-hydroxyl-E-coumaroyl)-D-glucosyl]-(1-2)-L-rhamnoside, 3-O-{2-O-[6-O-(p-hydroxyl-E-coumaroyl)-glucosyl]}-(1-2) rhamnosyl kaempfero were separated and identified as the major antioxidants. There is a linear relationship between the total amount of major antioxidants and total antioxidative activity of SI with a linear correlation coefficient of 0.9456. All the Relative standard deviations of recovery, precision and stability were below 7.5%. Based on these results, these nine ingredients could be selected as combinatorial markers to evaluate quality control of SI. It was concluded that on-line ABTS+-CE-DAD method was a simple, reliable and powerful tool to screen and quantify active ingredients for evaluating quality of herbal medicines.