Dietary supplementation of n-3 PUFAs ameliorates LL37-induced rosacea-like skin inflammation via inhibition of TLR2/MyD88/NF-κB pathway.

Rosacea is a facial chronic inflammatory skin disease with dysfunction of immune and neurovascular system and treatments for rosacea are challenging. N-3 polyunsaturated fatty acids (PUFAs), one of essential fatty acids, are needed for health maintenance and exert anti-inflammation and immunomodulatory effects in a series of cutaneous diseases such as atopic dermatitis and photoaging through dietary supplementation. However, the role of n-3 PUFAs on rosacea remains to be elucidated. In this study, KEGG enrichment analysis and GO analysis indicated that the biological process and signaling pathways, including chemokine signaling pathway, regulated by n-3 PUFAs highly overlapped with those in the pathogenic biological process of rosacea, especially the erythema telangiectasia type. Next, mice were randomized to fed with a customized n-3 PUFAs diet. We showed that n-3 PUFAs ameliorated skin erythema, inhibited dermal inflammatory cell infiltration (mast cells, neutrophils, and CD4 +T cells) and suppressed elevated pro-inflammatory cytokines in LL37-induced rosacea-like mice. Besides, n-3 PUFAs were also verified to repress angiogenesis in LL37-induced mice skin. Further investigation revealed that n-3 PUFAs attenuated LL37-induced inflammation via TLR2/ MyD88/ NF-κB pathway both in mice and in keratinocytes. In conclusion, our findings underscore that dietary supplementation of n-3 PUFAs have the potential to become an efficient and safe clinical therapeutic candidate for rosacea.

[Rapid screening and identification of 167 illegally added medicines in herbal tea by ultra high performance liquid chromatography-electrostatic field orbitrap high resolution mass spectrometry].

An accurate mass database and a method based on ultra high performance liquid chromatography-electrostatic field orbitrap high resolution mass spectrometry (UHPLC-Orbitrap HRMS) were developed. These were applied in the screening and identification of illegally added medicines in herbal tea. Based on investigations, 167 medicines were selected to build an accurate MS database; these medicines included antipyretic analgesics, glucocorticoids, antibiotics, and antihistamines, among other categories. The database was established using Orbitrap HRMS and TraceFinder software. The database carried information on all selected compounds, including the molecular formula, accurate mass of precursor ions and fragment ions, retention time, and mass spectra. The samples were ultrasonically extracted with a 50% (v/v) methanol aqueous solution. The extracted solutions were separated using a Waters XBrigde BEH C18 column (100 mm×2.1 mm, 2.5 µm). As the mobile phases, 0.1% (v/v) formic acid aqueous solution and acetonitrile containing 0.1% (v/v) formic acid were used, with gradient elution. The sample solutions were analyzed by Orbitrap HRMS in the full-scan MS and data-dependent MS/MS acquisition modes (Full MS/dd-MS2). Positive and negative polarity data were simultaneously acquired. Some parameters were optimized to increase the peak intensity and sensitivity of all compounds. The resolutions in the full-MS scan and dd-MS2 scan were set to 70000 and 17500, respectively. In the full-MS mode, scanning was performed in the range of m/z 100 to 1000. In the MS/MS mode, the normalized collision energy (NEC) was set to 20%, 40%, and 60% for each compound. The inclusion list was not used during the measurement, and the dynamic exclusion time was set to 10.0 s. The loop count was set to 5. After acquiring the sample data with these conditions using Orbitrap MS, they were imported into TraceFinder software, through which the sample information was extracted and automatically matched with the information on compounds in the MS database. Screening and identification were conducted by comparing the retention times as well as the exact masses of precursor ions and fragment ions that were experimentally measured. If the errors between the experimentally and theoretically obtained masses of the precursor ions were below 5×10-6 and the deviations in retention times were less than 20 s, then suspicious positive compounds might be identified. Furthermore, if such compounds possess more than one similar fragment ion with a mass tolerance below 5×10-6, and exhibit similar ion distributions in the MS/MS profiles (compared to those in the database), they could be confirmed to be the same. The validation result showed that all compounds had good linear relationships, with correlation coefficients (r) greater than 0.99. Because pefloxacin, norfloxacin, desloratadine, astemizole and clindamycin had background interference, the method was not suitable for their quantification. Following experiments using three spiked concentrations, the recoveries of the rest 162 compounds were found to be in the range of 66.4%-118.1%, and the relative standard deviations (RSDs, n=6), in the range of 0.1%-16.1%. When the limit of detection (LOD) was 0.2 mg/kg, 83 compounds were detected, while when the LOD was 1.0 mg/kg, 167 compounds were detected. All compounds were matched successfully to the standard added sample with the MS database in TraceFinder software. To lower the likelihood of false positive and false negative results, a quality control method was recommended. The method was applied to analyze 245 herbal tea samples, among which 12 positive samples were detected. Thirteen positive compounds were found, including acetaminophen, diclofenac sodium, chlorpheniramine, brompheniramine, dexamethasone, dexamethasone 21-acetate, prednisone, prednisone 21-acetate, metronidazole, erythromycin, ciprofloxacin, amantadine, and dextromethorphan. In particular, amantadine, dextromethorphan, brompheniramine, and ciprofloxacin were newly detected, compared to standard methods. The developed method is rapid and accurate, and will be useful in the high-throughput screening of illegally added medicines in herbal tea.

Comparison of red light and blue light therapies for mild-to-moderate acne vulgaris: A randomized controlled clinical study.

BACKGROUND: Red and blue light therapies are safe and effective treatments for mild-to-moderate acne vulgaris. However, very few previous studies have directly compared the characteristics of these two methods. OBJECTIVE: To compare the efficacy and side effects of red light (RL) and blue light (BL) for acne vulgaris and to assess these two therapies in different types of lesions. MATERIALS AND METHODS: A total of 28 subjects with mild-to-moderate acne vulgaris were randomized into the RL group or the BL group. Subjects in each group received different light treatments, and they were followed up regularly until 2 weeks after the last treatment. The improvement rates of different types of acne lesions were compared between the 2 groups, as well as the incidence of adverse reactions. RESULTS: At the 2-week follow-up, the average improvement rate of total acne lesions was 36.2% in the RL group and 30.7% in the BL group (p > .05). The average improvement rate of inflammatory and non-inflammatory lesions was 51.5% and 17.3% in the RL group, compared with 26.4% and 10.0% in the BL group (all p > .05). Treatment-related adverse reactions were observed distinctly in the BL group. CONCLUSIONS: Red light and BL therapies have similar efficacy in mild-to-moderate acne vulgaris, especially for inflammatory lesions. RL had advantages with fewer adverse reactions compared with BL.

Proteomics and lipidomics reveal the protective mechanism of dietary n-3 PUFA supplementation for photoaging.

Chronic ultraviolet radiation exposure could induce photoaging, and even carcinogenesis. Dietary omega-3 polyunsaturated fatty acid (n-3 PUFA) supplementation has proved to alleviate photoaging and cutaneous carcinoma. Although the exact mechanism remains poorly elucidated, accumulated evidence suggests that the alleviation effect of n-3 PUFA for photoaging is a multifactorial procession characterized by different pathways. Here, we performed a whole-genome proteomics and lipidomics analyses using a self-constructed photoaging mouse model with n-3 PUFA or n-6 PUFA supplementation. Significant alleviation of photoaging was observed, and a total of 88 differentially expressed proteins and 152 differentially expressed lipids were identified in mice with n-3 PUFA supplementation. We found that n-3 PUFA may alleviate photoaging by upregulating Hmmr (hyaluronic acid receptor) expression, which can decrease Mmp9 expression, reducing collagen degradation. As most proteins were associated with lipogenesis and lipid metabolism, we further analyzed the lipidomics data, finding that most triglycerides (93%) showed a significant increase in the n-3 PUFA supplementation group. Our proteomics and lipidomics results indicate that the protective mechanism of n-3 PUFA for photoaging is complicated. Furthermore, the effect of elevated triglycerides by n-3 PUFA supplementation in counteracting skin photoaging cannot be ignored, which will become a new prime target in anti-photoaging.

[Determination of chloramphenicol in propolis and propolis-derived dietary supplements by high performance liquid chromatography-tandem mass spectrometry].

A method for determining chloramphenicol (CAP) in both propolis and propolis-derived dietary supplements was developed by utilizing high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The flavones in the samples were removed with a lead acetate solution and ammonia, and the fat-soluble interferences, such as beewax and vegetable oils, were removed with n-hexane after the sample dissolved in ethanol. Tert-butyl methyl ether was used as the back-extraction solvent to reduce co-extracting compounds, such as polyethylene glycol 400 (PEG 400) and glycerol, which are common adjuvants of dietary supplements, and some polar interferences. CAP was detected by HPLC-MS/MS and quantified by the internal standard method. The calibration curve showed a good linearity in the range of 0.20-50.0 µg/L. The limits of detection and the limits of quantification were 0.03 and 0.1 µg/kg, respectively. The recoveries in four different matrices at three spiked levels were in the 86.0%-114.4% range with the relative standard deviations from 0.3% to 4.9%. With the advantages of excellent universality, ease of operation, high sensitivity, and strong anti-interference capability, the proposed method was suitable for the determination of CAP in both propolis and propolis-derived dietary supplements.

[Determination of 28 exogenous medicines and endogenous components in herbal drink using high performance liquid chromatography].

A high performance liquid chromatographic method for the determination of 28 exogenous medicines and endogenous components in the herbal drink was developed. The samples were extracted ultrasonically with methanol-water (70:30, v/v), and the extracts were separated in a Thermo Accucore C18 column (100 mm×4.6 mm, 2.6 µm) with methanol-acetonitrile-20 mmol/L ammonium acetate solution (pH 4.2) as the mobile phases by gradient elution. The flow rate was 1.2 mL/min and the column temperature was 35℃. The detection wavelengths were 254 nm and 220 nm. Quantification analysis was performed by the external standard method. The result showed the compounds had a good linear relationship in the range of 1-100 mg/L, and the correlation coefficients (r) were not less than 0.999. The limits of detection (LODs) of the 28 compounds were 1-10 mg/kg in the liquid sample and 20-200 mg/kg in the solid sample. The average recoveries of the 28 compounds in the liquid and solid samples were in the ranges of 88.8%-118.6% and 92.7%-112.3% with the relative standard deviations (RSDs) of 0.1%-6.7% and 0.1%-6.4%, respectively. The method was applied to analyze 456 herbal drink samples, and 55 positive samples were found. The positive rate was 12.1%. The developed method was simple and reliable, and it was suitable for the determination of 28 components in the herbal drink.

Arctigenin Attenuates Ischemia/Reperfusion Induced Ventricular Arrhythmias by Decreasing Oxidative Stress in Rats.

BACKGROUND/AIMS: Arctigenin (ATG) has been shown to possess anti-inflammatory, immunemodulatory, anti-viral, anti-microbial, anti-carcinogenic, vasodilatory and anti-platelet aggregation properties. However, the protective role of ATG in prevention of arrhythmias induced by myocardial ischemia/reperfusion is unknown. The aim of this study was to investigate the anti-arrhythmia effect of ATG in an ischemia/reperfusion injured rat heart model and explore the related mechanisms. METHODS: Rats were randomly exposed to sham operation, myocardial ischemia/ reperfusion (MI/R) alone, ATG+ MI/R, pretreated with ATG in low (12.5 mg/kg/day), medium (50 mg/kg/day) and high dose (200 mg/kg/day), respectively. Ventricular arrhythmias were assessed. The activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the level of malondialdehyde (MDA) in myocardial tissue were determined by chemical analysis. RESULTS: Compared to MI/R, rats pretreated with ATG in doses of 50 mg/kg/day and 200 mg/kg/day showed significantly reduced incidence and duration of ventricular fibrillation, ventricular tachycardia and ventricular ectopic beat (VEB), and decreased the arrhythmia score during the 30-min ischemia. Incidence and duration of ventricular tachycardia, infarction size and arrhythmia scores in these groups were significantly decreased during the 120-min reperfusion. No ventricular fibrillation occurred during the period of reperfusion. Rats pretreated with ATG in doses of 50 mg/kg/day and 200 mg/kg/ day markedly enhanced the activities of antioxidant enzymes SOD and GSH-Px, reduced the level of MDA. No differences were observed between the group pretreated with a low dose of ATG and the sham group. Administration of ATG significantly increased the expression of antioxidant stress protein Nrf2, Trx1 and Nox1. CONCLUSION: Our data suggested that ATG plays anti-arrhythmia role in ischemia/reperfusion injury, which is probably associated with attenuating oxidative stress by Nrf2 signaling pathway.