Sesquiterpenoids and steroids from Eupatorium fortunei and their inhibitory effects on NO production.

Two heterodimers including a clovane-phenylpropanoid hybrid (1) and a clovane-menthane hybrid (2), five linear sesquiterpenoids incorporating a tetrahydrofuran ring (3-6 & 8), and four steroids (7 & 9-11), were separated from the ethanolic extract of a well-known aromatic and medicinal herb Eupatorium fortunei. Their structures were characterised by detailed analyses of spectroscopic data and comparison with known analogues, with seven (1-7) of them being described for the first time. The hybrids 1 and 2 represent the first examples of clovane type sesquiterpenoids hybridising with other class of natural products, and compounds 3-6 and 8 are first linear sesquiterpenyl constituents reported from the title species. All the isolates were evaluated for their inhibitory effect on the NO production induced by LPS in murine RAW264.7 macrophage cells, and 1, 7, 10 and 11 exhibited moderate activity with IC50 values in the range of 24.4-43.5 µM.

Bioactive monoterpenoids and acetophenones from the aerial parts of Eupatorium fortunei.

Thirty-nine thymol and acetophenone derivatives, including eight pairs of enantiomers, were isolated from the aerial parts of Eupatorium fortunei. Their structures were assigned by detailed analyses of spectroscopic data and NMR calculations based on density functional theory, with 18 ones (1a/1b-14) being previously undescribed compounds. While the absolute configurations of 1a/1b, 2a/2b, 4, 6a/6b, 7, 11a/11b and 15a/15b-18a/18b were established by calculations of electronic circular dichroism data, that of 14 was determined by modified Mosher's method. Compounds 1a/1b and 2a/2b represent a previously unreported type of monoterpenoid dimers via an amide linkage, and compound 3 is a monoterpene-phenylpropanoid hybrid connected through an ester bond. Among the known molecules, the formerly mis-assigned structures of 15a/15b and 22 were revised, and pure natural enantiomers of 16a/16b-18a/18b were reported for the first time. Selective compounds showed antiradical and NO production inhibitory activities in the preliminary biological screening. Compound 31 was further demonstrated to alleviate oxidative stress by activating Nrf2 signaling pathway.

Chemical constituents from Eupatorium fortunei and their anti-inflammatory evaluation by in silico and experimental approaches.

The well-known aromatic and medicinal plant Eupatorium fortunei Turcz. is widely cultivated in China, and previous studies on its bioactive constituents mainly focus on the essential oil ingredients especially thymol derivatives. However, reports on other type of constituents and the potential application are lacking. In the present project, an intensive chemical fractionation on the aerial part extract of E. fortunei led to the isolation and identification of a series of fatty acid derivatives (lipids, 1a/1b-19) including seven pairs of previously undescribed enantiomers (1a/1b-7a/7b), as well as a lignan (brachangobinan A (BBA), 20) and two monoterpenes (8S/8R-9-hydroxythymol, 21a/21b). A preliminary biological evaluation of these compounds in a NO production inhibitory assay model demonstrated compound BBA as the most active one. Network pharmacology analysis was used to predict and explore the possible anti-inflammatory targets and mechanism of BBA, which revealed some potential inflammation-related proteins and signaling pathways. Further experimental investigations validated that the anti-inflammatory effect of BBA could be achieved by suppressing pro-inflammatory factors and blocking the activation of NF-κB signaling pathway. Taken together, our work shows that E. fortunei can serve as a potential resource of lipids and anti-inflammatory agents.

Anti-inflammatory labdane diterpenoids from the aerial parts of Leonurus japonicus.

Twenty-two labdane-type diterpenoids, including ten pairs of 15-epimers and a pair of 13,15-epimers, were obtained from the aerial parts of a well-known medicinal plant Leonurus japonicus Houtt. While these epimers were separated by chiral HPLC, their structures were established mainly via spectroscopic methods especially NMR, X-ray crystallography and ECD techniques. Among them, seventeen compounds, encompassing three pairs of solvolysis artefacts likely due to the use of ethanol as extracting solvent, were reported for the first time in the current work. Our preliminary anti-inflammatory screening demonstrated that seven diterpenoids displayed noteworthy inhibitory effect on the NO production in LPS-induced RAW264.7 cells. In addition, the release of pro-inflammatory factors TNF-α, IL-1ß and IL-6, as well as the expression of iNOS and COX-2 proteins, was also suppressed by the unreported 15,16-epoxy-6ß-hydroxy-15α-methoxy-7,16-dioxolabd-8,13-diene. Further investigation into the preliminary anti-inflammatory mechanism of this compound indicated that it could block the activation of NF-κB signaling pathway.

Inhalation of Cananga odorata essential oil relieves anxiety behaviors in autism-like rats via regulation of serotonin and dopamine metabolism.

OBJECTIVE: Anxiety is one of the most common symptoms associated with autistic spectrum disorder. The essential oil of Cananga odorata (Lam.) Hook. f. & Thomson, usually known as ylang-ylang oil (YYO), is often used in aromatherapy as a mood-regulating agent, sedative, or hypotensive agent. In the present study, the effects and mechanisms of YYO in alleviating anxiety, social and cognitive behaviors in autism-like rats were investigated. METHODS: The prenatal valproic acid (VPA) model was used to induce autism-like behaviors in offspring rats. The effectiveness of prenatal sodium valproate treatment (600 mg/kg) on offspring was shown by postnatal growth observation, and negative geotaxis, olfactory discrimination and Morris water maze (MWM) tests. Then three treatment groups were formed with varying exposure to atomized YYO to explore the effects of YYO on the anxiety, social and cognitive behaviors of the autistic-like offspring through the elevated plus-maze test, three-chamber social test, and MWM test. Finally, the monoamine neurotransmitters, including serotonin, dopamine and their metabolites, in the hippocampus and prefrontal cortex (PFC) of the rats were measured using a high-performance liquid chromatography. RESULTS: Offspring of VPA exposure rats showed autism-like behaviors. In the VPA offspring, medium-dose YYO exposure significantly elevated the time and entries into the open arms in the elevated plus-maze test, while low-dose YYO exposure significantly enhanced the social interaction time with the stranger rat in session 1 of the three-chamber social test. VPA offspring treated with YYO exposure used less time to reach the platform in the navigation test of the MWM test. YYO exposure significantly elevated the metabolism of serotonin and dopamine in the PFC of VPA offspring. CONCLUSION: YYO exposure showed the effects in alleviating anxiety and improving cognitive and social abilities in the offspring of VPA exposure rats. The role of YYO was related to the regulation of the metabolism of serotonin and dopamine. Please cite this article as: Zhang N, Wang ST, Yao L. Inhalation of Cananga odorata essential oil relieves anxiety behaviors in autism-like rats via regulation of serotonin and dopamine metabolism. J Integr Med. 2023; 21(2): 205-214.

Should ocular Demodex be checked and treated in refractory keratitis patients without blepharitis?

AIM: To evaluate the correlation between Demodex infestation and keratitis, and to assess demodicosis using a simple approach. METHODS: A modified slit lamp illumination (at 40× magnification) was used to observe Demodex tails in 40 patients with refractory keratitis and 80 healthy controls. Bacterial smear and culture of the conjunctival sac and corneal lesion were performed to identify the pathogen. Tea tree oil ointment (TTOO) was added as a Demodex killing agent for lid scrubs to the treatment when Demodex infestation was confirmed. RESULTS: Demodex tails were found in all patients compared to 42/80 of the controls (P<0.01). Seventeen patients presented blepharitis, while 23 were free of scales and inflammation at the lid margin. The demodicosis was mild, moderate, and severe in 8, 19, and 13 patients, respectively, compared to mild in 42 controls (P<0.01). The keratitis was mild, moderate, and severe in 13, 19, and 8 patients, respectively. The severity of Demodex infestation was not correlated to the severity of keratitis (P=0.126). The growth of Staphylococcus was revealed in nine patients who did not react to antibiotic eye drops prior to the TTOO treatment. Patients' signs and symptoms got resolved after the lid scrub with TTOO. CONCLUSION: Ocular Demodex needs to be checked and treated in refractory keratitis patients with or without blepharitis. A slit-lamp illumination under high magnification favors the judgment of the severity of Demodex infestation.

[Effects of manure combined with nitrogen fertilizer on yield and quality of winter wheat and soil biological characteristics in drylands of Northwest China]. / æœ‰æœºè‚¥é æ–½æ°®è‚¥å¯¹è¥¿åŒ—æ—±åœ°å†¬å°éº¦äº§é‡ã€å“è´¨åŠåœŸå£¤ç”Ÿç‰©å­¦ç‰¹æ€§çš„å½±å“.

We conducted an experiment with five treatments in winter wheat of the dryland of Northwest China, i.e. 30 t·hm-2 cow dung (M) plus different doses of nitrogen fertilizer (0, 75, 150, 225, and 300 kg N·hm-2), denoted by M+N0, M+N75, M+N150, M+N225, and M+N300, respectively. After three years of treatment, wheat yield, grain quality, and soil biological characteristics were measured in two consecutive years (2018 and 2019). The results showed that the combination of manure with nitrogen fertilizer significantly increased wheat yield compared with the manure-only treatment (M+N0). Compared with the manure-only treatment, the combined treatments significantly increased wheat grain protein content, wet gluten, sedimentation value, and extensibility, but not for starch content. Neither wheat yield nor grain quality had significant differences among the M+N150, M+N225, and M+N300 treatments, but both were prominently higher than those of M+N75. Soil microbial biomass carbon (MBC) and nitrogen (MBN) reached highest in M+N150 for both years, which were distinctly higher than those of M+N0, M+N225, and M+N300. In 2018, soil ß-1, 4-glucosidase, cellobiohydrolase, L-leucine aminopeptidase, ß-1,4-N-acetyl glucosaminidase, and alkaline phosphatase activities in M+N150 treatment were higher than those of other treatments. In 2019, soil enzyme activities (excluding L-leucine aminopeptidase) in M+N150 were higher than those of M+N0 and M+N225. MBC significantly positively correlated with MBN, and both significantly positively correlated with the activities of cellobiohydrolase, ß-1, 4-N-acetyl glucosaminidase, and alkaline phosphatase. MBN significantly positively correlated with total nitrogen content and negatively correlated with NO3-. Considering winter wheat yield, grain quality, and soil biological characteristics, M+N150 was conducive to sustainable production of winter wheat in drylands of Northwest China.

Integrated spatially resolved metabolomics and network toxicology to investigate the hepatotoxicity mechanisms of component D of Polygonum multiflorum Thunb.

ETHNOPHARMACOLOGICAL RELEVANCE: The liver toxicity of Reynoutria multiflora (Thunb.) Moldenke. (Polygonaceae) (Polygonum multiflorum Thunb, PM) has always attracted much attention, but the related toxicity materials and mechanisms have not been elucidated due to multi-component and multi-target characteristics. In previous hepatotoxicity screening, different components of PM were first evaluated and the hepatotoxicity of component D [95% ethanol (EtOH) elution] in a 70% EtOH extract of PM (PM-D) showed the highest hepatotoxicity. Furthermore, the main components of PM-D were identified and their hepatotoxicity was evaluated based on a zebrafish embryo model. However, the hepatotoxicity mechanism of PM-D is unknown. AIM OF THE STUDY: This work is to explore the hepatotoxicity mechanisms of PM-D by integrating network toxicology and spatially resolved metabolomics strategy. MATERIALS AND METHODS: A hepatotoxicity interaction network of PM-D was constructed based on toxicity target prediction for eight key toxic ingredients and a hepatotoxicity target collection. Then the key signaling pathways were enriched, and molecular docking verification was implemented to evaluate the ability of toxic ingredients to bind to the core targets. The pathological changes of liver tissues and serum biochemical assays of mice were used to evaluate the liver injury effect of mice with oral administration of PM-D. Furthermore, spatially resolved metabolomics was used to visualize significant differences in metabolic profiles in mice after drug administration, to screen hepatotoxicity-related biomarkers and analyze metabolic pathways. RESULTS: The contents of four key toxic compounds in PM-D were detected. Network toxicology identified 30 potential targets of liver toxicity of PM-D. GO and KEGG enrichment analyses indicated that the hepatotoxicity of PM-D involved multiple biological activities, including cellular response to endogenous stimulus, organonitrogen compound metabolic process, regulation of the apoptotic process, regulation of kinase, regulation of reactive oxygen species metabolic process and signaling pathways including PI3K-Akt, AMPK, MAPK, mTOR, Ras and HIF-1. The molecular docking confirmed the high binding activity of 8 key toxic ingredients with 10 core targets, including mTOR, PIK3CA, AKT1, and EGFR. The high distribution of metabolites of PM-D in the liver of administrated mice was recognized by mass spectrometry imaging. Spatially resolved metabolomics results revealed significant changes in metabolic profiles after PM-D administration, and metabolites such as taurine, taurocholic acid, adenosine, and acyl-carnitines were associated with PM-D-induced liver injury. Enrichment analyses of metabolic pathways revealed tht linolenic acid and linoleic acid metabolism, carnitine synthesis, oxidation of branched-chain fatty acids, and six other metabolic pathways were significantly changed. Comprehensive analysis revealed that the hepatotoxicity caused by PM-D was closely related to cholestasis, mitochondrial damage, oxidative stress and energy metabolism, and lipid metabolism disorders. CONCLUSIONS: In this study, the hepatotoxicity mechanisms of PM-D were comprehensively identified through an integrated spatially resolved metabolomics and network toxicology strategy, providing a theoretical foundation for the toxicity mechanisms of PM and its safe clinical application.

Yin-Yang: Programming Abstractions for Cross-Domain Multi-Acceleration.

FPGA accelerators offer performance and efficiency gains by narrowing the scope of acceleration to one algorithmic domain. However, real-life applications are often not limited to a single domain, which naturally makes Cross-Domain Multi-Acceleration a crucial next step. The challenge is, existing FPGA accelerators are built upon their specific vertically-specialized stacks, which prevents utilizing multiple accelerators from different domains. To that end, we propose a pair of dual abstractions, called Yin-Yang, which work in tandem and enable programmers to develop cross-domain applications using multiple accelerators on a FPGA. The Yin abstraction enables cross-domain algorithmic specification, while the Yang abstraction captures the accelerator capabilities. We also develop a dataflow virtual machine, dubbed XLVM, that transparently maps domain functions (Yin) to best-fit accelerator capabilities (Yang). With six real-world cross-domain applications, our evaluations show that Yin-Yang unlocks 29.4× speedup, while the best single-domain acceleration achieves 12.0×.

UHPLC-QQQ-MS/MS assay for the quantification of dianthrones as potential toxic markers of Polygonum multiflorum Thunb: applications for the standardization of traditional Chinese medicines (TCMs) with endogenous toxicity.

BACKGROUND: The raw and processed roots of Polygonum multiflorum Thunb (PM) are commonly used in clinical practice to treat diverse diseases; however, reports of hepatotoxicity induced by Polygoni Multiflori Radix (PMR) and Polygoni Multiflori Radix Praeparata (PMRP) have emerged worldwide. Thus, it is necessary for researchers to explore methods to improve quality standards to ensure their quality and treatment effects. METHODS: In the present study, an ultra-high performance liquid chromatography triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS) method was optimized and validated for the determination of dianthrones in PMR and PMRP using bianthronyl as the internal standard. Chromatographic separation with a gradient mobile phase [A: acetonitrile and B: water containing 0.1% formic acid (v/v)] at a flow rate of 0.25 mL/min was achieved on an Agilent ZORBAX SB-C18 column (2.1 mm × 50 mm, 1.8 µm). The triple quadrupole mass spectrometer (TQMS) was operated in negative ionization mode with multiple reaction monitoring for the quantitative analysis of six dianthrones. Moreover, compounds 5 and 6 were further evaluated for their cytotoxicity in HepaRG cells by CCK-8 assay. RESULTS: The UHPLC-QQQ-MS/MS method was first developed to simultaneously determine six dianthrones in PMR and PMRP, namely, polygonumnolides C1-C4 (1-4), trans-emodin dianthrones (5), and cis-emodin dianthrones (6). The contents of 1-6 in 90 batches of PMR were in the ranges of 0.027-19.04, 0.022-13.86, 0.073-15.53, 0.034-23.35, 0.38-83.67 and 0.29-67.00 µg/g, respectively. The contents of 1-6 in 86 batches of commercial PMRP were in the ranges of 0.020-13.03, 0.051-8.94, 0.022-7.23, 0.030-12.75, 0.098-28.54 and 0.14-27.79 µg/g, respectively. Compounds 1-4 were almost completely eliminated after reasonable processing for 24 h and the contents of compounds 5 and 6 significantly decreased. Additionally, compounds 5 and 6 showed inhibitory activity in HepaRG cells with IC50 values of 10.98 and 15.45 µM, respectively. Furthermore, a systematic five-step strategy to standardize TCMs with endogenous toxicity was proposed for the first time, which involved the establishment of determination methods, the identification of potentially toxic markers, the standardization of processing methods, the development of limit standards and a risk-benefit assessment. CONCLUSION: The results of the cytotoxicity evaluation of the dianthrones indicated that trans-emodin dianthrones (5) and cis-emodin dianthrones (6) could be selected as toxic markers of PMRP. Taking PMR and PMRP as examples, we hope this study provides insight into the standardization and internationalization of endogenous toxic TCMs, with the main purpose of improving public health by scientifically using TCMs to treat diverse complex diseases in the future.

Monochloropropanediol in edible vegetable oils from Hangzhou market in China: occurrence and exposure risk assessment.

In this study, 241 vegetable-oil food samples were collected from the Hangzhou market in China and analysed for fatty acid esters of 3- and 2-monochloropropanediol (3-MCPD and 2-MCPD) using non-derivative gas chromatography tandem mass spectrometry (GC-MS/MS). Food consumption data were taken from a food consumption survey of urban and rural residents in Hangzhou city performed in 2010-2011. Levels of 3-MCPD esters in edible oil ranged from not detected to 7.98 mg/kg, and the highest mean levels were found in tea seed oil, with concentrations of 2.94 mg/kg. Esters of 2-MCPD levels ranged from not detected to 4.03 mg/kg, and the highest mean levels were also found in tea seed oil, containing 1.49 mg/kg. The range of mean dietary intake of 3-MCPD esters in different groups of edible oil was from 0.096 to 1.54 µg/kg body weight (bw) per day, which is lower than the tolerable daily intake (TDI) established by the European Food Safety Authority (EFSA) (2 µg/kg bw/day). For people aged above 6 years old, the dietary intake of 3-MCPD from edible oil was 0.42 µg/kg bw per day (mean) and 1.22 µg/kg bw per day (P97.5). The range of mean dietary intake of 2-MCPD esters in different groups of edible oil was from 0.025 to 0.79 µg/kg bw/day, and 2-MCPD esters intake was 0.20 µg/kg bw per day (mean) and 0.60 µg/kg bw per day (P97.5). In addition, the dietary intake exposure to 3-MCPD and 2-MCPD esters for urban residents was lower than that for rural residents. The findings indicate that the potential health risks caused by dietary 3-MCPD esters from edible oils were of low concern for most of the Hangzhou residents. However, the exposure risk for consumers with excessive consumption of certain kind of edible oil calls for attention.

[Occurrence Characteristics and Health Risk Assessment of Endocrine Disrupting Chemicals in Groundwater in Wuxi-Changzhou].

The concentrations of 22 endocrine disrupting chemicals, including 4 categories of sex hormones, progesterones, glucocorticoids, and phenolic hormones, in groundwater of Wuxi-Changzhou were analyzed using solid-phase extraction and ultra-performance liquid chromatography tandem mass spectrometry. The results indicated that 20 EDCs were detected, among which bisphenol A (BPA), estrone (E1), estradiol (E2), and 17α-ethinyl estradiol (EE2) were the main components with detection ratios greater than 90%, followed by estriol (E3) and methyl-prednisolonel (Meprednl), which had detection rates 67.7% and 48.4%, respectively. The average concentration of BPA was the highest (4.95 ng·L-1), followed by that of Meprednl, E2, prednisolone (Prednl), testosterone propionate (TES-pro), and megestrol-acetate (MA; 2.84, 0.71, 0.64, 0.58, and 0.53 ng·L-1, respectively). The total concentration in each site ranged from 0.38 to 147.35 ng·L-1. The sites with higher concentration were mainly distributed in the Xinbei District, Liyang City, and the Zhonglou District of Changzhou City. The potential sources were analyzed by a principal component analysis (PCA) combining the point sources near sample sites. The results showed that domestic sewage, aquaculture, and industrial wastewater were the main sources, following by medical wastewater. A non-carcinogenic risk assessment showed a low risk of EDCs in the groundwater of the study area, and the risk of drinking water was greater than that of skin exposure. EE2 had the highest non-carcinogenic risk and is suggested to be monitored in priority.

In vivo hepatotoxicity screening of different extracts, components, and constituents of Polygoni Multiflori Thunb. in zebrafish (Danio rerio) larvae.

Polygonum multiflorum Thunb. (PM) is a traditional Chinese medicine, commonly used to treat a variety of diseases. However, the hepatotoxicity associated with PM hampers its clinical application and development. In this study, we refined the zebrafish hepatotoxicity model with regard to the following endpoints: liver size, liver gray value, and the area of yolk sac. The levels of alanine aminotransferase, aspartate transaminase, albumin, and microRNAs-122 were evaluated to verify the model. Subsequently, this model was used to screen different extracts, components, and constituents of PM, including 70 % EtOH extracts of PM, four fractions from macroporous resin (components A, B, C, and D), and 19 compounds from component D. We found that emodin, chrysophanol, emodin-8-O-ß-D-glucopyranoside, (cis)-emodin-emodin dianthrones, and (trans)-emodin-emodin dianthrones showed higher hepatotoxicity compared to other components in PM, whereas polyphenols showed lower hepatotoxicity. To the best of our knowledge, this study is the first to identify that dianthrones may account for the hepatotoxicity of PM. We believe that these findings will be helpful in regulating the hepatotoxicity of PM.

Fully automated identification and quantification of five polar pesticides in groundwater by isotope dilution-online solid phase extraction coupled with high-performance liquid chromatography-quadrupole Orbitrap high-resolution mass spectrometry.

A fully automated method for identification and quantification of five polar pesticides in groundwater by isotope dilution-online solid-phase extraction (SPE) coupled with high-performance liquid chromatography-quadrupole Orbitrap high-resolution mass spectrometry was developed. After one step of filtration, an aliquot of a 7.5-ml water sample was automatedly preconcentrated and purified on a turbulent Cyclone SPE column. The analytes were eluted in backflush mode, then separated on an analytical column and acquired by full MS/dd-MS2 scan in negative and positive ions mode. The major parameters for sample loading, cleanup, and elution were optimized in detail. Preconcentration and ionization efficiency were highly improved by using 0.1% acid solution in the mobile phase. The method provided good linearity of calibration coefficients (R2 > 0.995), sensitive method limits of detection (0.5-10.0 ng/L), accurate mass spectra (within 5 ppm error), satisfactory matrix spiking recoveries (98.4% to 109%), and high precision (intraday/interday relative standard deviations 1.57-8.90%). The method was successfully applied to analyze large batch groundwater of National Groundwater Monitoring Project and suspect screening of potential pesticides in groundwater. The study provided a practical alternative for a simple, robust, sensitive, and accurate identification and qualification of five polar pesticides in groundwater.

Determination of phthalate esters in breast milk before and after frozen storage in milk storage bags.

This study describes the migration of 20 phthalates from breast milk storage bags to breast milk after storage frozen for 1 to 6 months. Analysis of the investigated phthalates was performed by GC-MS/MS. Nine phthalates were found in some of the 27 breast milk storage bags, six phthalates were detected in some of the 40 breast milk samples, but the concentrations were low, except for one sample, which contained dibutyl phthalate (DBP) at the level of 211.2 µg/kg. For most of the breast milk samples stored in breast milk storage bags, the migration of diisobutyl phthalate (DIBP), DBP and di-ethyl-hexyl phthalate (DEHP) increased with increasing storage time, but the migration of DBP and DEHP was lower than the migration limits of 300 µg/kg and 1500 µg/kg from the National Standards of the PRC even after storage for six months. The average daily intakes of DMP, DEP, DIBP, DBP and DEHP were 0.099, 0.39, 0.62, 2.89 and 1.13 µg/kg/d, respectively, and the hazard indices were far below 1.

A limbic circuitry involved in emotional stress-induced grooming.

Prolonged exposure to negative stressors could be harmful if a subject cannot respond appropriately. Strategies evolved to respond to stress, including repetitive displacement behaviours, are important in maintaining behavioural homoeostasis. In rodents, self-grooming is a frequently observed repetitive behaviour believed to contribute to post-stress de-arousal with adaptive value. Here we identified a rat limbic di-synaptic circuit that regulates stress-induced self-grooming with positive affective valence. This circuit links hippocampal ventral subiculum to ventral lateral septum (LSv) and then lateral hypothalamus tuberal nucleus. Optogenetic activation of this circuit triggers delayed but robust excessive grooming with patterns closely resembling those evoked by emotional stress. Consistently, the neural activity of LSv reaches a peak before emotional stress-induced grooming while inhibition of this circuit significantly suppresses grooming triggered by emotional stress. Our results uncover a previously unknown limbic circuitry involved in regulating stress-induced self-grooming and pinpoint a critical role of LSv in this ethologically important behaviour.

Physical activity intervention for non-diabetic patients with non-alcoholic fatty liver disease: a meta-analysis of randomized controlled trials.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is currently the most common cause of chronic liver disease nowadays. Changes in diet and lifestyle have led to a dramatic increase in the prevalence of NAFLD around the world. This meta-analysis is to investigate the efficacy of physical activity intervention on liver-specific endpoints in the population with NAFLD, including hepatic enzyme, serum lipid, glucose metabolism and intra-hepatic lipid. METHODS: PubMed and China National Knowledge Infrastructure (CNKI) databases were searched for randomized clinical trials of physical activity intervention on NAFLD patients through April 20th, 2019. Effect sizes were reported as standardized mean difference (SMD) and 95% confidence intervals (CI). Quality of included studies was assessed according to the Cochrane risk of bias tool. Meta-analyses were conducted using random-effect or fixed-effect models depending on the significance of heterogeneity. Subgroup analyses according to types and duration of physical activity were conducted to investigate clinical variability. RESULTS: Nine studies with a cumulative total of 951 participants met selection criteria. Physical activity was found associated with small reductions in hepatic enzyme parameters: ALT (SMD -0.17, 95% CI:-0.30 to - 0.05), AST (SMD -0.25, 95% CI: - 0.38, - 0.13) and GGT (SMD -0.22, 95% CI: - 0.36, - 0.08). Significant small improvements were also found in serum lipid parameters including TC (SMD = - 0.22, 95% CI: - 0.34, - 0.09), TG (SMD = - 0.18, 95% CI: - 0.31 to - 0.06) and LDL-C (SMD = - 0.26, 95% CI: - 0.39 to - 0.13). Significant improvement was also found in intra-hepatic lipid content (SMD = - 0.21, 95% CI: - 0.36 to - 0.06) There was no difference between physical intervention group and control group in HDL and three glucose metabolism parameters. Subgroup analysis suggested both aerobic exercise alone and resistance exercise alone can improve most liver function and longer period of exercise generally had better improvement effect. CONCLUSIONS: Our findings suggest that physical activity alone can only slightly improve hepatic enzyme levels, most serum lipid levels and intra-hepatic lipid content in non-diabetic patients with NAFLD.

Estradiol-17ß regulates proliferation and apoptosis of sheep endometrial epithelial cells by regulating the relative abundance of YAP1.

Yes-associated protein 1 (YAP1) transcription regulator of the Hippo protein kinase pathway, serves as a key regulator of tissue growth and organ size by regulating cell proliferation and apoptosis. Effects of YAP1 on proliferation and apoptosis of sheep endometrial epithelial cells (EEC) as a result of estradiol-17ß (E2) treatment, however, remain unclear. In the present study, the abundance of YAP1 protein in the uterine horn was greater than that in the uterine body or cervix. The YAP1 protein was primarily localized in the endometrial luminal and glandular epithelial cells of the uterine horn of ewes on day 2 of the estrous cycle. Compared with control samples, there was a lesser abundance of YAP1 mRNA transcript that was associated with a lesser proliferation and greater apoptosis of EEC. There were also lesser concentrations of epidermal growth factor and insulin-like growth factor 1 in the spent culture medium when there was a lesser abundance of YAP1 mRNA in EEC compared with those in the control group. When there was a greater abundance of YAP1 mRNA transcript, there were greater concentrations of epidermal growth factor and insulin-like growth factor 1 in the spent media. Furthermore, with estradiol-17ß treatment the abundance of YAP1 mRNA transcript was similar to that of the control samples. Taken together, estradiol-17ß may function as an essential regulator of EEC proliferation and apoptosis by modulation of concentrations of YAP1 protein in the sheep uterus. These results indicate there are molecular mechanisms of estradiol-17ß and YAP1 in EEC proliferation and apoptosis of ewes.

Determination of vanadium, chromium, nickel, selenium and arsenic in calcium based toothpastes by microwave digestion-ICP-MS/MS / 预防医学

Objective@#To establish analytical method for the determination of vanadium ( V ), chromium ( Cr ), nickel ( Ni ), selenium ( Se ) and arsenic ( As ) in calcium based toothpastes by microwave digestion-inductively coupled plasma tandem mass spectrometry ( ICP-MS/MS ).@*Methods@#The 21 calcium based toothpaste samples from supermarkets and shops in the urban areas of Hangzhou were digested by 6 mL HNO3 and 1 mL H2O2 in microwave digestion system. Then He-SQ mode and O2-MS/MS mode of ICP-MS/MS were respectively used for the determination of Ni and V, Cr, Se, As. Indium ( In ) was used as internal standard for calibration. @*Results@# Good linear relationships were obtained for these five elements from 1.0 to 32.0 μg/L, with the correlation coefficients ranged from 0.999 3 to 1.000 0. The detection limits of the method ranged from 0.000 25 to 0.006 08 mg/kg. The recovery rates of standard spiking were 80.7%-105.7% when set at 0.4, 0.8 and 1.6 mg/kg, the recovery of standard reference material was 102.2%, and the relative standard deviations were 2.6%-4.8%. The concentrations of V, Cr, Ni, Se and As in 21 calcium based toothpaste samples were 0.024-1.935 mg/kg, 0.085-5.759 mg/kg, 0.090-3.673 mg/kg, <0.002 72-0.016 mg/kg and <0.006 08-0.321 mg/kg.@*Conclusion@#Microwave digestion-ICP-MS/MS can effectively reduce the interferences of polyatomic ions and doubly charged ions from the matrix, which is suitable for the determination of V, Cr, Ni, Se and As in calcium based toothpaste.

Detection of 20 phthalate esters in breast milk by GC-MS/MS using QuEChERS extraction method.

A detection method for 20 different phthalate esters (PAEs) in breast milk analyzed by quick, easy, cheap, effective, rugged, and safe (QuEChERS) clean-up with five internal standards and quantitative GC-MS/MS was established. This method can effectively remove interfering substances such as lipids and fatty acids from breast milk using acetonitrile as the extraction solvent and PSA/C18 as clean-up materials. The 20 PAEs had a linear range of 5.0-500.0 µg/L and recoveries of 83.3-123.3% with RSDs of 0.2-7.6% (n = 6). The method detection limits (LODs) were 0.004-1.3 µg/kg. Dimethyl phthalate (DMP), Diethyl phthalate (DEP), Diisobutyl phthalate (DIBP), Di-n-butyl phthalate (DBP) and bis(2-ethylhexyl) phthalate (DEHP) were detected in the 35 breast milk samples, with median levels of 1.2, 6.8, 7.3, 32.9 and 13.6 µg/kg, respectively, and the concentrations of 20 PAEs ranged between 0.5 and 137.3 µg/kg. This is a fast, simple, sensitive and accurate method for the detection of 20 PAEs in breast milk or dairy products.