Role of phosphoglucomutase in regulating trehalose metabolism in Nilaparvata lugens.

Phosphoglucomutase (PGM) is a key enzyme in glycolysis and gluconeogenesis, regulating both glycogen and trehalose metabolism in insects. In this study, we explored the potential function of phosphoglucomutase (PGM) using RNA interference technology in Nilaparvata lugens, the brown planthopper. PGM1 and PGM2 were found highly expressed in the midgut of brown planthoppers, with different expression levels in different instar nymphs. The glycogen, glucose, and trehalose levels were also significantly increased after brown planthoppers were injected with dsRNA targeting PGM1 (dsPGM1) or PGM2 (dsPGM2). In addition, injection of dsPGM1 or dsPGM2 resulted in increased membrane-bound trehalase activity but not soluble trehalase activity. Furthermore, the expression of genes related to trehalose and glycogen metabolism decreased significantly after injection with dsPGM1 and dsPGM2. The expression levels of genes involved in chitin metabolism in the brown planthopper were also significantly decreased and the insects showed wing deformities and difficulty molting following RNAi. We suggest that silencing of PGM1 and PGM2 expression directly inhibits trehalose metabolism, leading to impaired chitin synthesis.

Review of Anemone raddeana Rhizome and its pharmacological effects.

The chemical compositions of Anemone raddeana Rhizome, a kind of traditional Chinese medicine, were reviewed, along with its bioactivity and pharmacological properties and method improvements of extracting and detecting triterpenoid saponins. A. raddeana Rhizome is used to treat neuralgia and rheumatism, and is rich in triterpenoid saponins, most of which are pentacyclic, with oleanane as the nucleus. So far, 37 triterpenoid saponins have been determined from the herb. Its reported bioactivity and pharmacological properties have been described as anticancerous, antimicrobial, anti-inflammatory, analgesic, antipyretic, anticonvulsive, antihistaminic, and sedative. It has also been used for the induction of the humoral immune response and treatment of liver fibrosis in chronic hepatitis. However, the herb also has hemolytic effects and can be toxic, which limits its clinical application. Further studies are needed on the pharmaceutical functions, mechanisms, and immunological responses to contribute to the herb's clinical applications.

[Study on the Fingerprint of Kingkong Zedoary Turmeric Oil].

OBJECTIVE: To study the fingerprint of Zedoary Turmeric Oil (ZTO) as the bulk drug of Kingkong Elemene for making it safe, effective, stable, and controllable. METHODS: Fingerprints were detected by gas chromatography. ß-elemene peak was regarded as reference peak (S). The relative peak area of each common peak and the relative retention time were calculated. With a total of modes for reference, the fingerprints of 10 batches of Kingkong ZTO were detected, and their similarity was calculated by traditional Chinese medicine (TCM) fingerprint similarity calculation software. RESULTS: The determination method was stable and reliable. Totally 19 common characteristic peaks of Kingkong ZTO was found. The fingerprint similarity of these batches of Kingkong ZTO were not lower than 0.96. CONCLUSIONS: Gas chromatography for detecting the fingerprint of Kingkong ZTO was reliable and repeatable. The established fingerprint of Kingkong ZTO could guarantee the quality stability and safety of different product batches.

Trace matrix solid phase dispersion using a molecular sieve as the sorbent for the determination of flavonoids in fruit peels by ultra-performance liquid chromatography.

A simple, rapid, and highly selective trace matrix solid phase dispersion (MSPD) technique, coupled with ultra-performance liquid chromatography-ultraviolet detection, was proposed for extracting flavonoids from orange fruit peel matrices. Molecular sieve SBA-15 was applied for the first time as a solid support in trace MSPD. Parameters, such as the type of dispersant, mass ratio of the sample to the dispersant, grinding time, and elution pH, were optimized in detail. The optimal extraction conditions involved dispersing a powdered fruit peel sample (25 mg) into 25mg of SBA-15 and then eluting the target analytes with 500 µL of methanol. A satisfactory linearity (r(2) > 0.9990) was obtained, and the calculated limits of detection reached 0.02-0.03 µg/mL for the compounds. The results showed that the method developed was successfully applied to determine the content of flavonoids in complex fruit peel matrices.

Effervescence and graphitized multi-walled carbon nanotubes assisted microextraction for natural antioxidants by ultra high performance liquid chromatography with electrochemical detection and quadrupole time-of-flight tandem mass spectrometry.

In this article, effervescence and graphitized multi-walled carbon nanotubes assisted microextraction was first developed for the extraction of antioxidants in hawthorn samples. The use of an effervescent tablet composed of sodium dihydrogen phosphate, sodium carbonate and micro-scale carboxyl graphitized multi-walled carbon nanotubes (extraction sorbent) was the core of the method. In this study, ultra high performance liquid chromatography coupled with electrochemical detection and quadrupole time-of-flight tandem mass spectrometry was performed for qualitative and quantitative analyses of target analytes in hawthorn foodstuffs. Several experimental factors, such as amount of effervescent salts, the sorbent, elution time and elution solvent, were systematically assessed. Under the optimized conditions, a good linearity with R values better than 0.9980 was obtained. The detection limits estimated at a signal-to-noise ratio of 3:1 were ranging from 0.01 to 0.18ng/mL. These results suggested that the proposed method could be an alternative and promising sample preparation tool in future food analysis.

Trace-chitosan-wrapped multi-walled carbon nanotubes as a new sorbent in dispersive micro solid-phase extraction to determine phenolic compounds.

This report describes the use of trace-chitosan-wrapped multi-walled carbon nanotubes (CS-MWCNTs) as a sorbent material in dispersive micro solid-phase extraction (DMSPE), which was combined with ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry to analyze phenolic compounds in chrysanthemum tea and a chrysanthemum beverage. In this study, for the first time, CS-MWCNTs were used as a sorbent for this microextraction mode. Moreover, the proposed method exhibits the advantages of simplicity, rapidity, small sample amount and ease of operation. Furthermore, all of the important parameters that affect the extraction efficiency, such as the sorbent, pH, extraction time and type of elution solvent, were investigated and optimized in the DMSPE. Under the optimized extraction condition, the limit of detection, which was calculated based on a signal-to-noise ratio of 3, was 0.22-16.19ngmL(-1). Satisfactory recovery values of 89-106% were obtained for the tested samples. The results show that the developed method was successfully applied to determine the content of chlorogenic acid and flavonoids in complex chrysanthemum samples.

Effervescent-salt-assisted dispersive micro-solid-phase extraction using mesoporous hybrid materials coupled with ultra-performance liquid chromatography for the determination of trace-level compounds in complicated plant preparations.

A novel effervescent-salt-assisted dispersive micro-solid-phase extraction using mesoporous hybrid materials was developed for the extraction of minute traces of constituents in complicated plant preparations. In this study, a special tablet containing carbon dioxide sources (sodium dihydrogenphosphate and sodium carbonate) and the sorbent (mesoporous hybrid materials) was prepared. The effects of different parameters influencing the extraction efficiency such as the concentration of salts, the type and concentration of mesoporous material, pH of diluent, and desorption solvents were investigated and optimized. Results show that the proposed method using green solvents (water) as extraction solutions required low consumption of sample amount and obtained high enrichment efficiency. Moreover, under optimized conditions, the tested tanshinones exhibited good linearity (r(2) > 0.994) in the concentration range of 0.5 to 80 ng mL(-1). The limits of detection values were lower than 0.0803 pg using UV-visible detection. The developed method was successfully applied for the analysis of trace tanshinones in compound Danshen dripping pill and Danqi tablet samples.

Highly sensitive analysis of flavonoids by zwitterionic microemulsion electrokinetic chromatography coupled with light-emitting diode-induced fluorescence detection.

A rapid zwitterionic microemulsion electrokinetic chromatography (ZI-MEEKC) approach coupled with light-emitting-diode-induced fluorescence (LED-IF, 480nm) detection was proposed for the analysis of flavonoids. In the optimization process, we systematically investigated the separation conditions, including the surfactants, cosurfactants, pH, buffers and fluorescence parameters. It was found that the baseline separation of the seven flavonoids was obtained in less than 5min with a running buffer consisting of 92.9% (v/v) 5mM sodium borate, 0.6% (w/v) ZI surfactant, 0.5% (w/v) ethyl acetate and 6.0% (w/v) 1-butanol. High sensitivity was obtained by the application of LED-IF detection. The limits of detection for seven flavonoids were in the range of 3.30×10(-8) to 2.15×10(-6)molL(-1) without derivatization. Ultimately, the detection method was successfully applied to the analysis of flavonoids in hawthorn plant and food products with satisfactory results.

[Effects of cyclic stretch on the induction of the transdifferentiation in human lung epithelial cells].

OBJECTIVE: To investigate the effect of mechanical stretch induced epithelial-mesenchymal transition in human lung epithelial cells BEAS-2B in vitro. METHODS: The human lung epithelial cells BEAS-2B were subjected to cyclic stretch by the FX-5000T system at 0.33 Hz of 10% or 20% elongation for 24, 48 and 72 hours respectively. The morphologic changes were observed by microscopy. The mRNA and protein expressions of E-cadherin, Cytokeratin-8 (CK-8), α-smooth muscle actin (α-SMA) and Vimentin were evaluated by immunofluorescence before and after mechanical stretch and fluorescent quantitation reverse transcription-polymerase chain reaction (qRT-PCR). RESULTS: (1) When stretch by 20% elongation for 48 hours, the morphological changes in BEAS-2B cells from cobblestone-like structure to elongated shape and obviously when stretch for up to 72 hours, while 10% elongation showed no significant morphological changes comparing to control. (2) Decreasing E-cadherin and CK-8 protein expression was associated with increased immunostaining for α-SMA protein at 72 hours after 20% mechanical stretch. (3) Expression of E-cadherin mRNA was decreased to 0.388±0.056 and 0.247±0.064 after 20% mechanical stretch for 48 hours and 72 hours compared with control without stretch (set 1, both P<0.05), expression of CK-8 mRNA was decreased to 0.436±0.060 at 72 hours after 20% mechanical stretch (P<0.01), α-SMA mRNA was increased to 1.437±0.267 (48 hours) and 1.261±0.247 (72 hours) after 20% mechanical stretch (both P<0.05), and Vimentin mRNA was increased to 1.679±0.172 (48 hours) after 20% mechanical stretch (P<0.05). Expression of E-cadherin mRNA was decreased to 0.387±0.081 at 72 hours after 10% mechanical stretch (P<0.05), Vimentin mRNA was increased to 1.688±0.179 at 48 hours after 10% mechanical stretch while other markers showed no significant changes comparing with control. CONCLUSIONS: Excessive mechanical stretch could induce epithelial-mesenchymal transition in lung epithelial cells BEAS-2B in vitro.

[Effects of cyclic stretch on expression of cytokines and intercellular adhesion molecule-1 in human pulmonary artery endothelial cell].

OBJECTIVE: To observe the effects of cyclic stretch on expression of cytokines and adhesion molecules in human pulmonary artery endothelial cells (HPAECs), herein to provide a theoretical basis to ventilator-induced lung injury (VILI). METHODS: HPAECs were subjected to cyclic stretch by the Flexcell FX-5000T system at 0.5 Hz of 10% or 20% elongation for 3, 6, 12, 24 hours respectively. The mRNA and protein expression of interleukin (IL-6, IL-8), monocyte chemotactic protein-1 (MCP-1) and intercellular adhesion molecule-1 (ICAM-1) was determined by fluorescent quantitation reverse transcription-polymerase chain reaction (qRT-PCR), enzyme linked immunosorbent assay (ELISA) or Western blotting. RESULTS: Increasing the stretch force, the mRNA and protein expression of IL-8, MCP-1, ICAM-1 were up regulated with increasing stretch time. Compared with the control (set 1), after 20% cyclic stretch for 24 hours, IL-8 mRNA expression was up regulated to 1.58±0.10, MCP-1 mRNA expression was up regulated to 2.85±0.52, and ICAM-1 mRNA expression was up regulated to 1.90±0.14 (all P<0.05). Compared with control group, after 20% cyclic stretch for 24 hours, the protein expression of IL-8 and MCP-1 in HPAEC was significantly increased (IL-8: 3401.08±439.60 ng/L vs. 1422.60±66.98 ng/L, MCP-1: 1117.64±237.54 ng/L vs. 307.88±80.84 ng/L, both P<0.05), ICAM-1 protein expression was up regulated to 2.15±0.40 (P<0.05), while the expression of IL-6 mRNA and protein had no statistic difference compared with control group. CONCLUSIONS: Cyclic stretch enhanced the expression of IL-8, MCP-1 and ICAM-1 in an intensity-dependent fashion, so it may be involved in the pathogenesis of lung injury induced by mechanical ventilation.