[Development of Self Training Device for Lung Function Compliance Guided by Meridians].

OBJECTIVE: To develop a self deep breathing training device which can improve lung function compliance and blood oxygen saturation. METHODS: The device consists of four parts:flow tube, measuring cylinder, mobile phone holder and meridian guidance audio-visual synthesis training software. The flow tube measures the flow rate of inhaled gas, the metering cylinder measures the total amount of inhaled gas, and the mobile phone rack is equipped with a mobile phone storing the meridian guidance audio-visual synthesis training software. RESULTS: The device is reasonable in structure and flexible in operation, which can meet the requirements of self deep inspiration training under the guidance of training module. CONCLUSIONS: Deep inspiration training under the guidance of guidance training module can form "deep and slow" abdominal breathing, and then improve lung function.

Microwave-assisted extraction followed by salting-out phase separation for hierarchical screening of illegal adulterants in aphrodisiac health products by multi-dimensional fingerprint profiling analysis.

A novel method for hierarchical screening of illegal adulterants in Fur seal ginseng pills (FSGP) products was developed by microwave-assisted extraction (MAE) coupled to salting-out assisted liquid-liquid extraction (SALLE) with multi-dimensional fingerprint profiling analysis. Using a homogeneous system formed by dimethyl carbonate (DMC) and water as the extractant, the MAE conditions were investigated to maximize extraction recoveries, followed by addition of ammonium sulfate to induce DMC phase separation for SALLE enrichment of 16 potentially illegal adulterants such as phosphodiesterase type-5 inhibitors, androgens, α receptor antagonists and yohimbine etc. By means of high-performance liquid chromatography (HPLC) with diode array detection (DAD) and fluorescence detection (FLD), multi-dimensional fingerprints were acquired by multi-wavelength detection to highlight the signals of the potentially illegal adulterants and reduce or remove interferences from the sample matrix. For high accuracy and reliability, a hierarchical screening strategy was designed by multi-dimensional fingerprinting profiling analysis (MDFPA). The method exhibited proper identification and quantification performance, and it was successfully applied to screening of illegal adulterants in 18 batches of the samples through the step-by-step MDFPA. Also, the results were further confirmed by ultra high-performance liquid chromatography-quadrupole-orbitrap mass spectrometry (UHPLC-Q-Orbitrap/MS). The proposed method was proved to be a green, efficient and reliable alternative to monitoring aphrodisiac health products.

Multi-dimensional fingerprint profiling analysis for screening and quantification of illegal adulterated antidiabetics in hypoglycemic health products by aqueous two-phase extraction and multi-wavelength detection.

A novel method for screening and quantification of illegal adulterated antidiabetics in hypoglycemic health products was developed by multi-dimensional fingerprint profiling analysis (MDFPA). By means of aqueous two-phase extraction (ATPE), using aqueous two-phase system (ATPS) of butanol-water as the extractant, 11 common antidiabetics could be effectively extracted to the upper and lower phases, respectively. HPLC separation conditions for the extracts from two phases were investigated by multi-wavelength detection before and after p-nitrobenzoyl chloride (p-NBC) and 2,4-dinitrofluorobenzene (DNBF) derivatizations to establish multi-dimensional fingerprints. For high accuracy and reliability, a hierarchical screening approach to screening illegal adulterated antidiabetics in samples was established by MDFPA and spectral purity examination. Meanwhile, detection limits of identification for illegal adulterants were defined by detection limits of spectra (SLOD). The proposed method exhibited good identification and quantification performances. SLODs, LODs and LOQs of 11 antidiabetics were 1.22-8.37 µg/g, 0.225-4.23 µg/g and 0.755-14.10 µg/g, respectively. They had good linearity ranged from 2.0 µg/g to 300.0 µg/g (R2 ≥ 0.9978). The recoveries and RSDs were 76.83-109.6% and 0.50-6.5%, respectively. The method was successfully applied to screening of 15 batches of samples in different forms. Among them, four samples were detected to contain 5.47 µg/g of metformin, 6.50 µg/g of phenformin, 3.69 µg/g of glibenclamide and 9.11 µg/g of glimepiride, respectively. The results proved that it was an efficient and feasible alternative to screening and detection of illegal adulterated antidiabetics in hypoglycemic health products.

TiO2/(K,Na)NbO3 Nanocomposite for Boosting Humidity-Sensing Performances.

Nanomaterials of TiO2, (K0.5Na0.5)NbO3, and the TiO2/(K0.5Na0.5)NbO3 nanocomposite were successfully synthesized by a hydrothermal method. Impedance-type humidity sensors were fabricated based on these materials. Our results reveal that the impedance of the TiO2/(K0.5Na0.5)NbO3 sensor changes by 5 orders of magnitude with an ultrahigh sensing response of Sf = 166 470 recorded at 100 Hz in the tested relative humidity (RH) range of 12-94%. This value is almost 2 and 4 orders of magnitude larger than that of the (K0.5Na0.5)NbO3 and TiO2 sensors, respectively. Interestingly, satisfactory response/recovery time (25/38 s, within 5 min), very small hysteresis (<5%), excellent stability, and good repeatability were also achieved in the TiO2/(K0.5Na0.5)NbO3 sensor. The improved sensing properties are ascribed to the synergistic effect of TiO2/(K0.5Na0.5)NbO3 heterojunction, which contributes the impedance that is susceptible to environmental humidity. This work underscores that it is a facile way to boost humidity-sensing performance by constructing proper nanocomposites.

Enantioseparation and determination of triazole fungicides in vegetables and fruits by aqueous two-phase extraction coupled with online heart-cutting two-dimensional liquid chromatography.

A novel method for determination of triazole fungicides and their enantiomers in foods was developed by aqueous two-phase extraction (ATPE) coupled with two-dimensional liquid chromatography (2D-LC). With an aqueous two-phase system (ATPS) of ethanol/K2HPO4 as biphasic extractant, the effects of the ATPS composition, extraction temperature and time on extraction yields of triazole fungicides were investigated. Optimum conditions were concluded as follows: ethanol concentration 31% (w/w) and K2HPO4 concentration 21% (w/w), extraction temperature 60 °C, extraction time 21 min. Based on reversed-phase separation systems, a 2D-LC was constructed by optimizing chromatographic conditions and online heart-cutting procedures. As a result, eight triazole fungicides and their enantiomers were separated respectively on C18 and cellulose-tris (3,5-dimethylphenylcarbamate) stationary phases, revealing good linear relationship in the range of 0.1218-30.06 µg/mL (R2 ≥ 0.9984). The detection limits and recoveries were 0.03113-0.3525 µg/mL and 71.57-107.8%, respectively. The ATPE-2D-LC method was successfully applied to chiral analysis of triazole fungicides in fruits and vegetables.

Qualitative and quantitative analysis of sesquiterpene lactones in Centipeda minima by UPLC-Orbitrap-MS & UPLC-QQQ-MS.

An ultra-high performance liquid chromatography coupled with high-resolution Orbitrap mass spectrometry (UPLC-Orbitrap-MS) method has been used to identify sesquiterpene lactones in the methanolic extract of Centipeda minima. Fifteen sesquiterpene lactones were tentatively identified based on retention time and accurate mass of external standards or exact accurate mass searching (within 2 ppm) by comparison of some previous isolated sesquiterpene lactones. Meanwhile, a rapid, sensitive, precise, and reliable ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-QQQ-MS) method has been developed to evaluate the quality of Centipeda minima through a simultaneous determination of five sesquiterpene lactones, namely brevilin A, arnicolide C, arnicolide D, microhelenin C, minimolide F. Chromatographic separation was achieved on a Waters Acquilty UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 µm) with a mobile phase consisting of a) 0.1% formic acid and b) a mixture of acetonitrile and methanol 50:50 v/v under an isocratic elution (42:58) manner. Positive electrospray ionization mode with multiple reaction monitoring was applied for the detection of the five sesquiterpene lactones. Method validation for linearity, accuracy and precision was also carried out. Finally, the method was successfully used for the analysis of 10 batches of Centipeda minima samples collected in China. Brevilin A and arnicolide D were the dominant sesquiterpene lactones in Centipeda minima and could be proposed as suitable markers for the quality control of Centipeda minima.

A hierarchical screening method for detection of illegal adulterants in Fur seal ginseng pills by profiling analysis of HPLC multi-dimensional fingerprints.

A novel method for hierarchical screening of illegal adulterants in Fur seal ginseng pills products was developed by multi-dimensional fingerprint profiling analysis. Fingerprint feature of the samples was acquired by high-performance liquid chromatography analysis of 11 batches of samples with diode array detector and fluorescence detector, and then potential illegal adulterants including phosphodiesterase type-5 inhibitors, androgens, α receptor antagonists and yohimbine, were further separated at multiple wavelengths to reduce or remove interferences from sample matrix for highlight their chromatographic characteristics. Accordingly, a hierarchical screening strategy was designed by first-order and second-order fingerprints combined with spectral examination to achieve high accuracy and reliability. The method was successfully applied to screening of illegal adulterants in real samples, and it also exhibited good quantification performance through validation tests. From 16 batches of samples, three suspected samples were confirmed to be positive, containing 9.37µg/g of testosterone, 18.8 µg/g of tadalafil, and 48.5 µg/g of sildenafil, respectively. The recoveries and relative standard deviations were in the range of 83.6-103.1% and 4.2-6.8%, respectively. The proposed method provided a simple, efficient and promising alternative to monitoring functional foods.

Combination of microwave-assisted extraction and ultrasonic-assisted dispersive liquid-liquid microextraction for separation and enrichment of pyrethroids residues in Litchi fruit prior to HPLC determination.

A novel method for simultaneous determination of pyrethroids residues in Litchi fruit has been developed by HPLC-UV detection using microwave-assisted extraction (MAE) coupled with ultrasonic-assisted dispersive liquid-liquid microextraction (UADLLME). Extraction conditions of MAE and UADLLME were respectively investigated by single-factor experiments and response surface methodology. Optimized experimental conditions included 310µL of chlorobenzene as extraction solvent, 1.3mL of ethanol as dispersive solvent and 3min of extraction time for UADLLME. In the case of MAE, extraction temperature of 70°C, extraction time of 4min and solvent-to-materials ratio of 40:1 were adopted. Results demonstrated that the proposed method had good performance with linearity of 0.0050-4.98mg/L, recovery of 83.3-91.5%, RSDs below 5.6% and detection limit (LOD) of 1.15-2.46µg/L for six pyrethroids, offering higher extraction efficiency and larger enrichment factor. MAE-UADLLME provided a sensitive and efficient alternative to determination of trace amounts of pesticides residues in food samples.

Ultra-Performance Liquid Chromatography Hyphenated with Quadrupole-Orbitrap Mass Spectrometry for Simultaneous Determination of Necine-Core-Structure Pyrrolizidine Alkaloids in Crotalaria sessiliflora L. without all Corresponding Standards.

INTRODUCTION: Crotalaria sessiliflora L. is a Chinese traditional herb for treatment of cutaneum carcinoma and cervical carcinoma. In addition to monocrotaline, coexisting pyrrolizidine alkaloids (PAs) also require further quantification for quality control and pharmaceutical uses of the herb. OBJECTIVE: To establish a UPLC-Q-Orbitrap/MS method of simultaneous determination of coexisting PAs with same parent structure for quality control and comprehensive researches of Crotalaria sessiliflora L. METHODOLOGY: PAs in Crotalaria sessiliflora L. were analysed by UPLC-Q-Orbitrap/MS method. Coexisting PAs were identified by mass data of full MS-dd-MS2 based on the characteristic fragmentation pattern and necine-core structure. Moreover, quantification of PAs was conducted by parallel reaction monitoring (PRM) mode using m/z 138, m/z 120 and m/z 94 from identical necine-core structure as quantitative ions with single monocrotaline standard for accurate calibration. RESULTS: Five PAs, named monocrotaline, retrorsine, senecionine, integerrimine, O-9-angeloylretronecine, were indentified and confirmed. Quantitative ions of m/z 138, m/z 120 and m/z 94 were used for quantification of PAs containing the necine-core structure in Crotalaria sessiliflora L. The results demonstrated that contents, precision and recoveries of the five PAs mentioned earlier were respectively 3.307-30.35 µg/g, 1.1-4.5% and 88.91-92.33% while using m/z 120 as the best quantitative ion. CONCLUSION: The UPLC-Q-Orbitrap/MS method was established for simultaneous determination of five PAs in Crotalaria sessiliflora L. without all corresponding standards, and was proved that it was simple, convenient and effective for comprehensive quality control and pharmaceutical uses. Copyright © 2017 John Wiley & Sons, Ltd.

Extraction and characterization of polysaccharides from Semen Cassiae by microwave-assisted aqueous two-phase extraction coupled with spectroscopy and HPLC.

A novel and rapid method for simultaneous extraction and separation of the different polysaccharides from Semen Cassiae (SC) was developed by microwave-assisted aqueous two-phase extraction (MAATPE) in a one-step procedure. Using ethanol/ammonium sulfate system as a multiphase solvent, the effects of MAATPE on the extraction of polysaccharides from SC such as the composition of the ATPS, extraction time, temperature and solvent-to-material ratio were investigated by UV-vis analysis. Under the optimum conditions, the yields of polysaccharides were 4.49% for the top phase, 8.80% for the bottom phase and 13.29% for total polysaccharides, respectively. Compared with heating solvent extraction and ultrasonic assisted extraction, MAATPE exhibited the higher extraction yields in shorter time. Fourier-transform infrared spectra showed that two polysaccharides extracted from SC to the top and bottom phases by MAATPE were different from each other in their chemical structures. Through acid hydrolysis and PMP derivatization prior to HPLC, analytical results by indicated that a polysaccharide of the top phases was a relatively homogeneous homepolysaccharide composed of dominant gucose glucose while that of the bottom phase was a water-soluble heteropolysaccharide with multiple components of glucose, xylose, arabinose, galactose, mannose and glucuronic acid. Molar ratios of monosaccharides were 95.13:4.27:0.60 of glucose: arabinose: galactose for the polysaccharide from the top phase and 62.96:14.07:6.67: 6.67:5.19:4.44 of glucose: xylose: arabinose: galactose: mannose: glucuronic acid for that from the bottom phase, respectively. The mechanism for MAATPE process was also discussed in detail. MAATPE with the aid of microwave and the selectivity of the ATPS not only improved yields of the extraction, but also obtained a variety of polysaccharides. Hence, it was proved as a green, efficient and promising alternative to simultaneous extraction of polysaccharides from SC.

A Systematic Analysis on mRNA and MicroRNA Expression in Runting and Stunting Chickens.

Runting and stunting syndrome (RSS), which is characterized by lower body weight, widely occurs in broilers. Some RSS chickens simply exhibit slow growth without pathological changes. An increasing number of studies indicate that broiler strains differ in susceptibility to infectious diseases, most likely due to their genetic differences. The objective of this study was to detect the differentially expressed miRNAs and mRNAs in RSS and normal chickens. By integrating miRNA with mRNA expression profiling, potential molecular mechanisms involved in RSS could be further explored. Twenty-two known miRNAs and 1,159 genes were differentially expressed in RSS chickens compared with normal chickens (P < 0.05). qPCR validation results displayed similar patterns. The differentially expressed genes were primarily involved in energy metabolism pathways. The antisense transcripts were extensively expressed in chicken liver albeit with reduced abundance. Dual-luciferase reporter assay indicated that gga-miR-30b/c directly target CARS through binding to its 3'UTR. The miR-30b/c: CARS regulation mainly occurred in liver. In thigh muscle and the hypothalamus, miR-30b/c are expressed at higher levels in RSS chickens compared with normal chickens from 2 to 6 w of age, and notably significant differences are observed at 4 w of age.

MicroRNA profile analysis on duck feather follicle and skin with high-throughput sequencing technology.

Skin acts an important protection role in animal survival and it evolves with the animal divergence. We identified the conserved miRNA families of skin among duck and other species. Cluster analysis showed that the species with similar skin characteristics were clustered into the same group, indicating miRNAs are important in skin function and skin evolution. The miRNA profiles demonstrated that different miRNA regulation mechanism may exist in contour feather follicles (with the surrounding skin) and down feather follicles (with the surrounding skin). Comparing the highly abundant miRNAs with those of mammalian hair follicles and skins, different miRNAs and miRNA families were found, suggesting the different ways in feather follicles and mammalian hair follicles. Bioinformatics prediction indicated that seven miRNAs probably targeted the genes of Wnt/ß-catenin, Shh/BMP and Notch pathways which were important in feather morphogenesis. Further analysis should be conducted to experimentally validate the relationships between miRNAs and their predicted target genes because the target genes were based exclusively upon the bioinformatics.

The GTPase activating Rap/RanGAP domain-like 1 gene is associated with chicken reproductive traits.

BACKGROUND: Abundant evidence indicates that chicken reproduction is strictly regulated by the hypothalamic-pituitary-gonad (HPG) axis, and the genes included in the HPG axis have been studied extensively. However, the question remains as to whether any other genes outside of the HPG system are involved in regulating chicken reproduction. The present study was aimed to identify, on a genome-wide level, novel genes associated with chicken reproductive traits. METHODOLOGY/PRINCIPAL FINDING: Suppressive subtractive hybridization (SSH), genome-wide association study (GWAS), and gene-centric GWAS were used to identify novel genes underlying chicken reproduction. Single marker-trait association analysis with a large population and allelic frequency spectrum analysis were used to confirm the effects of candidate genes. Using two full-sib Ningdu Sanhuang (NDH) chickens, GARNL1 was identified as a candidate gene involved in chicken broodiness by SSH analysis. Its expression levels in the hypothalamus and pituitary were significantly higher in brooding chickens than in non-brooding chickens. GWAS analysis with a NDH two tail sample showed that 2802 SNPs were significantly associated with egg number at 300 d of age (EN300). Among the 2802 SNPs, 2 SNPs composed a block overlapping the GARNL1 gene. The gene-centric GWAS analysis with another two tail sample of NDH showed that GARNL1 was strongly associated with EN300 and age at first egg (AFE). Single marker-trait association analysis in 1301 female NDH chickens confirmed that variation in this gene was related to EN300 and AFE. The allelic frequency spectrum of the SNP rs15700989 among 5 different populations supported the above associations. Western blotting, RT-PCR, and qPCR were used to analyze alternative splicing of the GARNL1 gene. RT-PCR detected 5 transcripts and revealed that the transcript, which has a 141 bp insertion, was expressed in a tissue-specific manner. CONCLUSIONS/SIGNIFICANCE: Our findings demonstrate that the GARNL1 gene contributes to chicken reproductive traits.

Study of acoustic field modulation in the regenerator by double loudspeakers method.

A model to modulate acoustic field in a regenerator of a thermoacoustic system by the double loudspeakers method is presented in this paper. The equations are derived for acoustic field modulation. They represent the relations among acoustic field (complex pressure p(0), complex velocity u(0), and acoustic impedance Z(0)), driving parameters of loudspeakers (voltage amplitude and its phase difference), and operating parameters involved in a matrix H (frequency, temperature of regenerator). The range of acoustic field is adjustable and limited by the maximal driving voltages of loudspeakers according to driving parameters. The range is simulated and analyzed in the amplitude-phase and complex coordinate planes for a given or variable H. The simulated results indicate that the range has its intrinsic characteristics. The expected acoustic field in a regenerator can be obtained feasibly by the modulation.

Investigation on the different types of resonators in the thermoacoustic Stirling prime mover.

A new thermoacoustic Stirling prime mover consisting of a looped tube and different types of resonators respectively has been built. Experiments were focused on three axisymmetric types of resonators: cylinder, cone and cylinder-cone under the same operation conditions. Pressure ratios in the prime mover were observed, 1.15 for cylinder-cone, 1.092 for cylinder and 1.097 for cone. The acoustic fields along the looped tube and different types of resonators were calculated, which agreed well with the measured results.

Design and experiment on a mini cascade thermoacoustic engine.

A miniature cascade thermoacoustic engine had been designed and tested, which length was about 1m, operating at 500 Hz. Pilot study and experimental results shown a rather good agreement between measured and calculated pressure fields and temperatures distributions in the thermoaoustic engine. The peak-to-peak value of the acoustic pressure was 0.02 MPa at the 1.8 MPa charged pressure of helium. Some efforts had been made to extend the traveling-wave region based on the analysis of the acoustic impedance.