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Acute altitude reaction is a stress response of organism to special altitude environmental factors such as hypoxia, low pressure, cold, dry and strong ultraviolet. As it is the most incident disease in high altitude areas, its prevention remains a problem to be solved. In China, the traditional Chinese (Tibetan) medicines have been recognized as an effective means of preventing and treating acute altitude sicknesses. Some single-recipe or compound traditional Chinese (Tibetan) medicines have been proved to be effective for acute altitude sicknesses. In this article, we will describe traditional Chinese (Tibetan) medicines of different types with efficacy in prevention and treatment of altitude sicknesses.;

Jikan Mingmu Drops (JMD), a traditional Tibetan medicine containing six herbs, has been used to treat dry eye syndrome (DES) in individuals with diabetes mellitus. However, the activity of JMD ameliorates DES with diabetes mellitus has not been previously examined. The aim of the study is to investigate the molecular mechanism of JMD on db/db mice. The main chemical constituents of JMD were analyzed by high-performance liquid chromatography and gas chromatography-mass spectrometry. DES was then induced in db/db mice by applying 0.2% benzalkonium chloride to the ocular surface for 7 days. Eye drops containing JMD (0.25, 0.5, or 1 g/mL) or vehicle subsequently were administered three times daily for another 7 days, and the therapeutic effects were evaluated by phenol red thread tear and sodium fluorescein tests. Conjunctival specimens were subjected to hematoxylin and eosin staining and periodic acid-Schiff staining to examine pathological changes and number of goblet cells. ELISA was performed to assess the levels of various inflammatory cytokines. JMD contains hydroxysafflor yellow A, magnoflorine, jatrorrhizine hydrochloride, palmatine hydrochloride, berberine hydrochloride, gallic acid, ellagic acid, tauroursodeoxycholic acid, camphor, isoborneol, borneol, trans-cinnamic acid, and muscone. JMD treatment significantly increased the tear volume, decreased the corneal fluorescein staining score, restored the morphology and structure of conjunctival epithelial cells, and markedly downregulated the levels of interleukin (IL)-6, IL-17α, IL-1β, tumor necrosis factor-α, and vascular endothelial growth factor in the conjunctiva. Further data showed that these protective effects were accompanied by inhibition of inflammation in a dose-dependent manner. Amelioration of DES in db/db mice with diabetes mellitus by treatment with Tibetan medicine formula JMD maybe related to its anti-inflammatory effects.

<br>Display Omitted<br>• 4′-Carbonyl chloride rosamine was synthesized and used for NTs by UHPLC-MS/MS. • <b>In situ</b> UA-DDLLME was reported for the simultaneous determination of AANTs and MANTs. • The method was sensitive, selective, low matrix effect, speedy and eco-friendly. • A new analytical tool in diagnosis of AD-related disease.<br>Neurotransmitters (NTs) may play an important role in neurodegenerative disorders such as Alzheimer’s disease (AD). In order to investigate the potential links, a new simple, fast, accurate and sensitive analytical method, based on <b>in situ</b> ultrasound-assisted derivatization dispersive liquid-liquid microextraction (<b>in situ</b> UA-DDLLME) coupled with ultra high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS), has been developed and validated. The quantitation of amino acid neurotransmitters (AANTs) and monoamine neurotransmitters (MANTs) in urine of AD rats were performed in this work. The <b>in situ</b> UA-DDLLME procedure involved the rapid injection of the mixture of low toxic 4-bromoanisole (extractant) and acetonitrile (dispersant), which containing the new designed and synthesized 4′-carbonyl chloride rosamine (CCR) as derivatization reagent, into the aqueous phase of real sample and buffer. Under the selected conditions, the derivatization and microextraction of analytes were simultaneously completed within 1 min. Good linearity for each analyte (R > 0.992) was observed with low limit of detections (LODs, S/N > 3). Moreover, the proposed method was compared with direct detection or other reported methods, and the results showed that low matrix effects and good recoveries results were obtained in this work. Taken together, <b>in situ</b> UA-DDLLME coupled with UHPLC-MS/MS analysis was demonstrated to be a good method for sensitive, accurate and simultaneous monitoring of AANTs and MANTs. This method would be expected to be highly useful in AD diseases’ clinical diagnostics and may have potential value in monitoring the efficacy of treatment.

The current study evaluated antioxidant activity of <i>Dracocephalum rupestre</i> Hance, characterized by the polyphenolic compounds in the ethyl acetate fraction (EAF) from <i>D. rupestre</i> and investigated the protective mechanisms of EAF in carbon tetrachloride (CCl₄)-induced hepatic injury. EAF showed the largest antioxidant capacity as demonstrated by DPPH, ABTS, OH, and FRAP assays (<i>p</i> ≤ .05). Using LC-MS, two polyphenolics, for example, rosmarinic acid, and eriodictyol were identified in EAF. Total phenol content of <i>D. rupestre</i> was correlated significantly with FRAP value (<i>r</i> = .999, <i>p</i> ≤ .001). The results also showed that EAF decreased serum ALT and AST activities compared with model group, as well as the histological findings. In addition, EAF remarkably decreased MDA and LDH levels in liver. This is the first time to reveal the hepatoprotective effect of EAF from <i>D. rupestre</i>, which may be developed as a new drug for treatment of liver injury in future.<br>Practical applications: <i>Dracocephalum rupestre</i> Hance is known as Chinese herbal medicine with pharmacological functions of antioxidant, anti-inflammatory and has been mainly used in the treatment of damp-heat, headache, fever, jaundice, hepatitis, liver toxicity, and other diseases. In order to detect the potential application of this plant, the antioxidant and hepatoprotective activities of <i>D. rupestre</i> were investigated. It was found that the ethyl acetate fraction (EAF) from <i>D. rupestre</i> displayed strong antioxidant activity. We also demonstrate that EAF could lessen CCl₄-induced acute liver injury in mice. Also, rosmarinic acid may play an important role in the antioxidant activity and hepatoprotective effect for EAF.

Swertia mussotii is a kind of rare medicinal materials, the relevant researches are mainly concentrated on its medicinal efficacy and medicinal value till now, researches of adaptive distribution by applying remote sensing and GIS are relatively less. This study is to analyze the adaptive distribution of S.mussotii in Sichuan province by applying remote sensing and GIS technology, and provide scientific basis for the protection and development of wild resources, artificial cultivation and adjustment of Chinese medicine industrial distribution in Sichuan province. Based on literature review and ecological factors such as altitude, annual precipitation and annual average temperature, this study extracted ecological factors, overlay analysis in GIS, as well as combining GPS field validation data by means of remote sensing and GIS, discusses the adaptive distribution of SMF sin Sichuan province. ①The area of adaptive distribution of S. mussotii in Sichuan province is 1 543.749 km², mainly in Dege county, Ganzi county, Daofu county, Kangding county, Barkam, Jinchuan county, Xiaojin county, Danba county, Daocheng county, Xiangcheng county, Xinlong county, Aba county, Muli county and other counties and cities, accounts for about 7.25% in total area. ② Combining statistical information and field validation, this study found that S. mussotii adaptive distribution gained by remote sensing and GIS is in conformity with its actual distribution. The study shows that remote sensing and GIS technology are feasible to obtain the S. mussotii adaptive distribution, they can further be applied to studies on adaptive distributions of other rare Chinese medicinal herb.

Abstract Juniperus pingii var. wilsonii has been traditionally used in Tibetan medicine for the treatment of inflammatory diseases. In the present study, J. pingii var. wilsonii polysaccharides (JPWP), with high content of d ‑galacturonic acid, showed potent anti-complementary activity in vitro and significantly attenuated acute lung injury (ALI) induced by H1N1 influenza virus in vivo through reducing the inflammatory responses, alleviating oxidative stress and inhibiting the activation of complement. Thus, anti-complementary activity-guided fractionation of JPWP led to the isolation of an acidic homogeneous polysaccharide, JPWP-PS, whose structure was further elucidated by acid hydrolysis, PMP derivation, methylation and NMR analysis. JPWP-PS had potent anti-complementary activity with the CH 50 value of 0.073 ± 0.009 mg/mL, and was characterized by the residues of T-Ara f -(1→, →3)-Ara f -(1→, →3,5)-Ara f -(1→, →3)-Gal p -(1→ and →4)-Gal p A-(1→. Graphical abstract Unlabelled Image Highlights • Juniperus pingii var. wilsonii polysaccharides (JPWP) improved survival rate of H1N1 virus infected mice. • JPWP treated acute lung injury via inhibiting inflammatory responses, oxidative stress and activation of complement. • A homogeneous acidic polysaccharide with potent anti-complementary activity was isolated from JPWP. [ABSTRACT FROM AUTHOR]

Background: Tai Chi is a traditional Chinese sport that is classified as a moderate exercise. Recent studies have evaluated the effectiveness of Tai Chi in substance abuse rehabilitation. Objectives: The aim of this study was to assess the quality of life and physical effects of a Tai Chi intervention on individuals with amphetamine-type stimulant (stimulant) dependence. Methods: Sixty male subjects with stimulant dependence from a Shanghai Mandatory Detoxification and Rehabilitation Center participated in a 12-week trial. Tai Chi was used as an intervention in the experimental group (n = 30). The control group (n = 29) underwent standard care, which included recreation activity, gesture language exercise, and self-education. Outcome measures included the quality of life for drug addiction (QOL-DA) questionnaire [four scales consisting of physiology (e.g., energy level), psychology (e.g., depression), symptoms (e.g., physical symptoms), society (e.g., interpersonal) and fitness evaluations (assessed by body mass index, body fat, hand-grip, flexibility, balance)]. Repeated measures were used to analyze the changes over time. Results: Test scores of the QOL-DA in the Tai Chi group significantly increased after 12 weeks in the following areas: physiology, 8.71 (p = 0.005), symptoms, 4.34 (p = 0.042), society, 15.79 (p < 0.001), and total score, 10.60 (p = 0.002). A post hoc test further revealed that quality of life improved in the Tai Chi group but not in the standard care group. Physical results showed a significant interaction with balance (F(1,56) = 6.92, p = 0.011); participants in the Tai Chi group improved by 10 s while there was no change in the standard care group. Although there were no significant interactions in the fitness outcomes (i.e., hand-grip and sit-and-reach tests), the within-group factor displayed significant changes in body fat (F(1,56) = 27.79, p < 0.001) in both groups. Conclusion: This study demonstrates that Tai Chi is a promising exercise that improves quality of life for individuals with stimulant dependence.

Seven phenolic compounds, <b>1</b> - <b>7</b>, including a new organic acid gallate, mucic acid 1-ethyl 6-methyl ester 2-<i>O</i>-gallate (<b>7</b>), were isolated from the MeOH extract of the fruits of <i>Phyllanthus emblica</i> L. (Euphorbiaceae). The structures were elucidated on the basis of extensive spectroscopic analysis and comparison with literature data. Upon evaluated for their antioxidant abilities by 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2′-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assays. The inhibitory activities against melanogenesis in B16 melanoma cells induced by <i>α</i>-MSH, as well as cytotoxic activities against four human cancer cell lines were also evaluated. All phenolic compounds, <b>1</b> - <b>7</b>, exhibited potent antioxidant abilities (DPPH:<i> IC</i><sub>50</sub> 5.6 - 12.9 μm; ABTS: 0.87 - 8.43 μm <i>Trolox</i>/μm; FRAP: 1.01 - 5.79 μm Fe<sup>2+</sup>/μm, respectively). Besides, <b>5</b> - <b>7</b>, also exhibited moderate inhibitory activities against melanogenesis (80.7 - 86.8% melanin content), even with no or low toxicity to the cells (93.5 - 101.6% cell viability) at a high concentration of 100 μm. Compounds <b>1</b> - <b>3</b> exhibited cytotoxic activity against one or more cell lines (<i>IC</i><sub>50</sub> 13.9 - 68.4%), and compound <b>1</b> with high tumor selectivity for A549 (<i>SI</i> 3.2).

A phytochemical investigation on the aerial parts of a Tibetan medicine Meconopsis horridula, by solvent extraction, repeated chromatographies on silica gel, Sephadex LH-20, and preparative TLC techniques, led to the isolation of 9 compounds. By spectroscopic analysis and comparison of its 1H and 13C-NMR data with those in literatures, their structures were identified as oleracein E(1), N-( trans-p-coumaroyl) tyramine (2), chrysoeriol (3), apigenin (4), hydnocarpin (5), p-coumaric acid glucosyl ester (6), stigmast-5-ene-3beta-ylformate (7), 3beta-hydroxy-7alpha-ethoxy-24beta-ethylcholest-5-ene (8), and beta-sitosterol (9), respectively, among which compounds 6-8 were isolated from the genus for the first time,and 1,3 were isolated from the species for the first time. A MTT method was applied to evaluate the cytotoxic activity of compounds 14 against the human hepatocellular liver carcinoma cell line (HepG2), and compound 1 showed significant cytotoxicity against HepG2,with its inhibitory rate of 52.2% at 10 micromol x L(-1).;

Graphical abstract The introduction of chiral enantiomers generated chiral β-HgS QDs, which showed chirality inversion compared with the corresponding chiral ligands. Their chiroptical activity, good cytocompatibility, near-infrared optical absorption, near-infrared fluorescence emission and high-performance photothermal conversion implied that these chiral β-HgS quantum dots have potential to be applied in biotechnology and bio-medicine. Abstract β-HgS quantum dots (QDs) have drawn enormous attention due to the size-tunable bandgap and the lowest quantum state in conduction band which have been applied to semiconductor transistor and photodetector. Though β-HgS is the essential component of Tibetan medicine, the potential toxicity of β-HgS limits its applications, especially in bio-application. Herein, chiral biomolecule enantiomers N -isobutyryl- L (D)-cysteine (L (D)-NIBC) and L (D)-cysteine (L (D)-Cys) were introduced into HgCl 2 and Na 2 S aqueous solution to synthesize chiral β-HgS QDs in one-pot, which significantly improved their water-solubility and cytocompatibility. Notably, all chiral β-HgS QDs showed none cytotoxicity even at high concentration (20 mg·L−1), and the cytocompatibility of D -β-HgS QDs was better than corresponding L -β-HgS QDs at the concentration of 20 mg·L−1. This cytotoxicity discrimination was associated with the chirality inversion of chiral β-HgS QDs compared with the corresponding chiral ligands. In-situ real-time circular dichroism (CD) monitoring indicated that the chirality of β-HgS QDs originated from the asymmetrical arrangement of chiral ligands on the achiral core surface. Their chiroptical activity, near-infrared optical absorption (800 nm), fluorescence emission (900–1000 nm), high-performance photothermal conversion and good cytocompatibility, implied chiral β-HgS QDs could be used as a candidate material for photothermal therapy or a near-infrared fluorescent probe in organism, which brings a novel insight for bio-application of β-HgS QDs. [ABSTRACT FROM AUTHOR]

One new compound, Colletotrichine A (1), was produced by the fungal Colletotrichum gloeosporioides GT-7. The structure was established by 1D and 2D NMR spectra. Monoamine oxidase (MAO) and acetylcholinesterase (AChE) inhibitory activity of 1 was also evaluated. Compound 1 showed AChE-inhibiting activity with IC50 value of 28 μg/mL.

OBJECTIVE: To further investigate and discuss the cause of species endangerment, the status and present problem of conservation of traditional Tibetan medicine in China.METHOD: Previous relevant investigations and literatures were summed up in the field. The present situation of conservation of traditional Tibetan medicine was analyzed. RESULT: The status of endangered resources, cause of species endangerment, the conserving status and conserving measures etc were elaborated. The classification was made and suggestion of species conservation of traditional Tibetan medicine were put forward. CONCLUSION: The endangered species conservation of traditional Tibetan medicine was carried out by building protective area of endangered species resources and plant garden, setting up germplasm bank, developing the domestication and cultivation of Tibetan medicinal herbs most in use, strengthening the investigation and study of endangered species, launching exchange and cooperation of conservation techniques on endangered species, enhancing the protective awareness of endangered species traditional Tibetan medicine etc. By so doing we can facilitate the sustainable development of traditional Tibetan medicine in China.

ETHNOPHARMACOLOGICAL RELEVANCE: Corydalis hendersonii Hemsl. (CH) with heat clearing and detoxifying effects are well described in Tibetan folk medicine. It has been used for centuries in China largely for the treatment of high altitude polycythemia, a pathophysiological condition referred to "plethora" in Tibetan medicine, hypertension, hepatitis, edema, gastritis, and other infectious diseases.AIM OF THE STUDY: To investigate the cardioprotective effects of Corydalis hendersonii extract in an ICR mouse model of myocardial ischemic injury. MATERIALS AND METHODS: Ethanol [85% (v/v)] extract of CH whole plant was prepared, and their chemical profile was analyzed with use of HPLC-DAD and IT-TOF-ESI-MS. A mouse model of AMI was established by ligation of the left ventricular dysfunction (LAD) coronary artery. Mice were randomly divided into six groups (n = 12 per group): sham group, model group, CH groups treated with three doses of CH (100, 200, and 400mg/kg, intragastric), and a positive control group (captopril, 16.67mg/kg, intragastric). Heart function was evaluated by measurement of ejection fraction (EF) and fractional shortening (FS) by echocardiography. Serum levels of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH), plasma levels of angiotensin II (AngII), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1ß (IL-1ß) and expressions of matrix metalloproteinase-2 (MMP-2) and MMP-9 in the cardiac tissue homogenate, protein expressions of signal-transduction proteins, p65, IκBα, JAK2, and STAT3 in heart tissues were measured by ELISA and Western blot analyses. Inflammatory cell infiltration and changes in collagen deposition in the myocardial ischemic heart tissues were observed by histopathological examination. Platelet aggregation in vitro was also assessed. RESULTS: CH treatment showed a dose-dependent cardioprotective effect. It significantly reduced left ventricular end-diastolic diameter (LVEDd) and left ventricular end-diastolic diameter (LVEDs), improved EF and FS as compared to those in the model group; attenuated the increase levels of CK-MB and LDH in serum; reduced expressions of AngII, TNF-α, IL-6 and IL-1ß in plasma, MMP-2 and MMP-9 expressions in the cardiac tissue homogenate; and down-regulated myocardial expressions of p-p65, p-IκBα, p-JAK2, p-STAT3, MMP-2, and MMP-9 in AMI mice. Also, an obvious reduction in inflammatory cell infiltration in the myocardial infarct was found in all CH treated groups. Besides, CH also inhibited platelet aggregation induced by THR, ADP, and AA. CONCLUSION: CH extract exerted a protective effect against myocardial ischemic injury via inhibition of inflammation, myocardial fibrosis, and platelet aggregation. This study demonstrates such protection for the first time and provides a basis for development of CH-based drugs for treatment of ischemic heart disease in clinical settings.

Rheum tanguticum Maxim. ex Balf. (Rt), a traditional Tibetan medicine, is known to exert various bioactivities, including anti-inflammatory and antioxidative activities. The present study was conducted to investigate anti-inflammatory and antioxidative effects of Rt on activated microglia. Rt (10 μg/ml) significantly inhibited the mean protein level of interleukin-1β (IL-1β) in the organotypic hippocampal slice cultures following treatment with chromogranin A (CGA, 10 nM) and pancreastatin (10 nM), endogenous microglial activators present in senile plaques. Rt also significantly inhibited the expression and production of inflammatory and oxidative molecules, including IL-1β, tumor necrosis factor-α, and nitric oxide, by cultured microglia after treatment with CGA. These effects of Rt are considered to be mediated by the secretion of interleukin-10 (IL-10) from microglia, because neutralizing antibodies against IL-10 significantly canceled these effects. To explore the causative components of Rt responsible for inducing the secretion of IL-10, the effects of seven components of Rt on the IL-10 expression in microglia were examined. Among them, aloe-emodin (10 μM) and (+)-catechin (30 μM) were able to induce the secretion of IL-10 from cultured microglia. Therefore, aloe-emodin and (+)-catechin are deemed responsible for the antineuroinflammatory and antioxidative effects of Rt through the secretion of IL-10 from microglia. Accordingly, Rt is considered potentially useful for the treatment of AD.

Rheum tanguticum Maxim. ex Balf. (Rt), a traditional Tibetan medicine, is known to exert various bioactivities, including anti-inflammatory and antioxidative activities. The present study was conducted to investigate anti-inflammatory and antioxidative effects of Rt on activated microglia. Rt (10 μg/ml) significantly inhibited the mean protein level of interleukin-1β (IL-1β) in the organotypic hippocampal slice cultures following treatment with chromogranin A (CGA, 10 nM) and pancreastatin (10 nM), endogenous microglial activators present in senile plaques. Rt also significantly inhibited the expression and production of inflammatory and oxidative molecules, including IL-1β, tumor necrosis factor-α, and nitric oxide, by cultured microglia after treatment with CGA. These effects of Rt are considered to be mediated by the secretion of interleukin-10 (IL-10) from microglia, because neutralizing antibodies against IL-10 significantly canceled these effects. To explore the causative components of Rt responsible for inducing the secretion of IL-10, the effects of seven components of Rt on the IL-10 expression in microglia were examined. Among them, aloe-emodin (10 μM) and (+)-catechin (30 μM) were able to induce the secretion of IL-10 from cultured microglia. Therefore, aloe-emodin and (+)-catechin are deemed responsible for the antineuroinflammatory and antioxidative effects of Rt through the secretion of IL-10 from microglia. Accordingly, Rt is considered potentially useful for the treatment of AD.

BACKGROUND:During the past decade, there has been a rapid increase of interactive apps designed for health and well-being. Yet, little research has been published on developing frameworks for design and evaluation of digital mindfulness facilitating technologies. Moreover, many existing digital mindfulness applications are purely software based. There is room for further exploration and assessment of designs that make more use of physical qualities of artifacts. OBJECTIVE: The study aimed to develop and test a new physical digital mindfulness prototype designed for stress reduction. METHODS: In this case study, we designed, developed, and evaluated HU, a physical digital mindfulness prototype designed for stress reduction. In the first phase, we used vapor and light to support mindful breathing and invited 25 participants through snowball sampling to test HU. In the second phase, we added sonification. We deployed a package of probes such as photos, diaries, and cards to collect data from users who explored HU in their homes. Thereafter, we evaluated our installation using both self-assessed stress levels and heart rate (HR) and heart rate variability (HRV) measures in a pilot study, in order to measure stress resilience effects. After the experiment, we performed a semistructured interview to reflect on HU and investigate the design of digital mindfulness apps for stress reduction. RESULTS: The results of the first phase showed that 22 of 25 participants (88%) claimed vapor and light could be effective ways of promoting mindful breathing. Vapor could potentially support mindful breathing better than light (especially for mindfulness beginners). In addition, a majority of the participants mentioned sound as an alternative medium. In the second phase, we found that participants thought that HU could work well for stress reduction. We compared the effect of silent HU (using light and vapor without sound) and sonified HU on 5 participants. Subjective stress levels were statistically improved with both silent and sonified HU. The mean value of HR using silent HU was significantly lower than resting baseline and sonified HU. The mean value of root mean square of differences (RMSSD) using silent HU was significantly higher than resting baseline. We found that the differences between our objective and subjective assessments were intriguing and prompted us to investigate them further. CONCLUSIONS: Our evaluation of HU indicated that HU could facilitate relaxed breathing and stress reduction. There was a difference in outcome between the physiological measures of stress and the subjective reports of stress, as well as a large intervariability among study participants. Our conclusion is that the use of stress reduction tools should be customized and that the design work of mindfulness technology for stress reduction is a complex process, which requires cooperation of designers, HCI (Human-Computer Interaction) experts and clinicians.

A novel hyphenated method based on ultrasound-assisted dispersive liquid-liquid microextraction coupled to precolumn derivatization has been established for the simultaneous determination of bisphenol A, 4-octylphenol, and 4-nonylphenol by high-performance liquid chromatography with fluorescence detection. Different parameters that influence microextraction and derivatization have been optimized. The quantitative linear range of analytes is 5.0-400.0 ng/L, and the correlation coefficients are more than 0.9998. Limits of detection for soft drinks and dairy products have been obtained in the range of 0.5-1.2 ng/kg and 0.01-0.04 μg/kg, respectively. Relative standard deviations of intra- and inter-day precision for retention time and peak area are in the range of 0.47-2.31 and 2.76-8.79%, respectively. Accuracy is satisfactory in the range of 81.5-118.7%. Relative standard deviations of repeatability are in the range of 0.35-1.43 and 2.36-4.75% for retention time and peak area, respectively. Enrichment factors for bisphenol A, 4-octylphenol, and 4-nonylphenol are 170.5, 240.3, and 283.2, respectively. The results of recovery and matrix effect are in the range of 82.7-114.9 and 92.0-109.0%, respectively. The proposed method has been applied to the determination of bisphenol A, 4-octylphenol, and 4-nonylphenol in soft drinks and dairy products with much higher sensitivity than many other methods.

A novel hyphenated technique based on ultrasonic-assisted dispersive liquid-liquid microextraction (UA-DLLME) coupled with derivatization has been established for the determination of brassinolide (BL, a representative of brassinosteroids) by HPLC fluorescence detection. 9-Phenanthreneboronic acid is used as labeling reagent of BL. UA-DLLME parameters containing type and volume of extraction and disperser solvent, pH and ultrasonication time are optimized. Derivatization parameters are optimized included amount of 9-phenanthreneboronic acid, volume ratio of pyridine, derivatization time and temperature. Under optimal conditions, quantitative linear range of BL is 50-1,000 ng L<sup>−1</sup> and excellent linear response is observed with correlation coefficient of 0.9996. Limit of detection and limit of quantification are calculated as 8.0 and 25.0 ng L<sup>−1</sup>, respectively. RSDs of retention time and peak area are in the range of 0.68-0.97 % and 4.61-6.54 % for intra-day precision, 1.32-1.94 % and 7.28-9.75 % for inter-day precision, respectively. Accuracy is satisfactory in the range of 82.3-125.1 %. RSDs’ values of repeatability are in the range of 0.82-1.79 and 3.95-8.53 % for retention time and peak area, respectively. Enrichment factor for BL is 189. The results of recovery and matrix effect are in the range of 82.0-108.6 and 90.0-115.3 %, respectively. The proposed method has been applied for the determination of BL in <i>Arabidopsis thaliana</i>, <i>Daucus carota</i> and <i>Brassica campestris</i> L. leaves with much higher sensitivity than many other methods.

PREMISE OF THE STUDY: Ephedragerardiana (Ephedraceae), occurring in the Himalayan ranges, is an important plant species used in Tibetan medicine. Due to the lack of molecular markers to characterize genetic diversity, knowledge for conservation and uses of E. gerardiana resources is limited; we therefore developed microsatellite markers for use in this species.METHODS AND RESULTS: Using Illumina MiSeq sequencing technology, we developed 29 polymorphic microsatellite loci suitable for E. gerardiana, of which 15 loci also showed polymorphisms in two related Ephedra species, E. saxatilis and E. monosperma. The average number of effective alleles per locus ranged from two to six. The observed and expected heterozygosity ranged from 0.23 to 0.83 and 0.44 to 0.86, respectively, in E. gerardiana populations. CONCLUSIONS: The developed 29 microsatellite markers are effective for the study of genetic structure and genetic diversity of E. gerardiana, and 15 of these markers are suitable for related Ephedra species.

PREMISE OF THE STUDY: Ephedragerardiana (Ephedraceae), occurring in the Himalayan ranges, is an important plant species used in Tibetan medicine. Due to the lack of molecular markers to characterize genetic diversity, knowledge for conservation and uses of E. gerardiana resources is limited; we therefore developed microsatellite markers for use in this species. METHODS AND RESULTS: Using Illumina MiSeq sequencing technology, we developed 29 polymorphic microsatellite loci suitable for E. gerardiana, of which 15 loci also showed polymorphisms in two related Ephedra species, E. saxatilis and E. monosperma. The average number of effective alleles per locus ranged from two to six. The observed and expected heterozygosity ranged from 0.23 to 0.83 and 0.44 to 0.86, respectively, in E. gerardiana populations. CONCLUSIONS: The developed 29 microsatellite markers are effective for the study of genetic structure and genetic diversity of E. gerardiana, and 15 of these markers are suitable for related Ephedra species.

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