Cafestol inhibits colon cancer cell proliferation and tumor growth in xenograft mice by activating LKB1/AMPK/ULK1-dependent autophagy.

Chemotherapy failure in colorectal cancer patients is the major cause of recurrence and poor prognosis. As a result, there is an urgent need to develop drugs that have a good chemotherapy effect while also being extremely safe. In this study, we found cafestol inhibited colon cancer growth and HCT116 proliferation in vivo and in vitro, and improved the composition of intestinal flora. Further metabolomic data showed that autophagy and AMPK pathways were involved in the process of cafestol's anti-colon cancer effects. The functional validation studies revealed that cafestol increased autophagy vesicles and LC3B-II levels. The autophagic flux induced by cafestol was prevented by using BafA1. The autophagy inhibitor 3-MA blocked the cafestol-induced increase in LC3B-II and cell proliferation inhibition. Then we found that cafestol induced the increased expressions of LKB1, AMPK, ULK1, p-LKB1, p-AMPK, and p-ULK1 proteins in vivo and in vitro. Using the siRNA targeted to the Lkb1 gene, the levels of AMPK, ULK1, and LC3B-II were suppressed under cafestol treatment. These results indicated that the effect of cafestol is through regulating LKB1/AMPK/ULK1 pathway-mediated autophagic death. Finally, a correlation matrix of the microbiome and autophagy-related proteins was conducted. We found that cafestol-induced autophagic protein expression was positively correlated with the beneficial intestinal bacteria (Muribaculaceae, Bacteroides, Prevotellacece, and Alloprevotella) and negatively correlated with the hazardous bacteria. Conclusions: This study found that cafestol inhibited colon cancer in vitro and in vivo by the mechanism that may be related to LKB1/AMPK/ULK1 pathway-mediated autophagic cell death and improved intestinal microenvironment.

Associations of three thermogenic adipokines with metabolic syndrome in obese and non-obese populations from the China plateau: the China Multi-Ethnic Cohort.

OBJECTIVES: High altitude exposure decreases the incidence of obesity and metabolic syndrome, but increases the expression of the thermogenic adipokines (leptin, fat cell fatty acid-binding protein (A-FABP) and visfatin). This study investigated the correlation of these adipokines with obesity and metabolic syndrome (MetS) in populations residing in a plateau-specific environment. DESIGN: Case-control study. SETTING: We cross-sectionally analysed data from the China Multi-Ethnic Cohort. PARTICIPANTS: A total of 475 obese (OB, body mass index (BMI)≥28.0 kg/m2) plateau Han people and 475 age, sex and region-matched non-obese (NO, 18.5≤BMI<24.0 kg/m2) subjects were recruited. MetS was defined according to the National Cholesterol Education Program Adult Treatment Panel III guidelines. PRIMARY AND SECONDARY OUTCOME MEASURES: Data with normal distributions were expressed as the mean (Stanard Deviation, SD), and data with skewed distributions were expressed as the median (Interquartile Range, IQR). The participants were grouped and the rank-sum test, χ2 test or t-tests was used for comparing groups. Spearman correlation coefficients were estimated to assess the relationships among leptin, A-FABP, visfatin and the components of MetS in each group. RESULTS: A-FABP was an independent predictor of OB (OR, 1.207; 95% CI, 1.170 to 1.245; p<0.05), ABSI (OR, 1.035; 95%CI, 1.019 to 1.052; p<0.05) and MetS (OR, 1.035; 95% CI, 1.013 to 1.057; p<0.05). Leptin was an independent predictor of MetS in the NO group. Visfatin was an independent predictor of increased ABSI, but not for OB or MetS. CONCLUSION: An abnormally elevated plasma A-FABP level, but not leptin or visfatin is a potential risk factor for MetS in high-altitude populations.

Plasma Fatty Acids, Not Dietary Fatty Acids, Associated with Obesity in Four Ethnic Minority Groups Unique to Southwest China: A Cross-Sectional Study.

Background: Dietary fatty acids (DFAs) and plasma fatty acids (PFAs) are linked to obesity. However, whether this association exists among ethnic minorities remains lacking. The present cross-sectional study was designed to investigate the correlation between DFAs, PFAs and obesity in four ethnic minority groups to Southwest China. Methods: A total of 166 obese people, and 166 normal-BMI subjects matched based on their age-, sex-, and ethnicity- were recruited from four different ethnic minority groups. DFAs were obtained through food frequency questionnaires. PFAs were assayed by GC/MS method. Binary and multiple regression analyses were performed to evaluate the correlation among DFAs, PFAs and obesity. Canonical correlation analysis (CCA) was conducted to assess the relationship between DFAs and PFAs. Results: FAs were found to be highest in the Naxi people and lowest in the Hani people. Multiple logistic regression analysis revealed that plasma C16:0 (OR = 1.310; 95% CI, 1.028-1.669) in the Hani people; plasma C20:3 n-6 (OR = 6.250; 95% CI, 1.224-31.927) and dietary C20:1 (OR = 9.231; 95% CI, 1.253-68.016) in the Wa people; plasma C18:0 (OR = 0.788; 95% CI, 0.681-0.912) in the Naxi people were seen to be independent predictive factors for obesity. CCA showed that DFAs were positively correlated with PFAs in the Naxi (r: 0.676; P < 0.05) and Bulang people (r: 0.897; P < 0.05), but there was no correlation in the Hani and Wa people. Conclusion: In this study, PFAs but not DFAs were independently associated with obesity, and different among the four ethnic minorities.

Rapid determination and dietary intake risk assessment of 249 pesticide residues in Panax notoginseng.

UPLC-MS/MS and GC-MS/MS were used to establish a method to simultaneously determine various pesticide residues in Panax notoginseng. Results showed that the limits of detection of 249 pesticides were all 5-10 µg/kg. The detection rate of pesticides in 121 P. notoginseng samples was 93.39%, and 19 pesticides were detected. According to the US Code of Federal Regulations, the Chinese Pharmacopoeia recommended algorithm, and the Japanese "positive list system", the pass rates of pesticide residues were 100%, 99.17%, and 89.26%, respectively. The chronic risk quotient (ADI%) and acute risk quotient (ARfD%) of P. notoginseng were 0.00-0.12% and 0.00-0.15%, respectively. In summary, the detection method established in this study can be used for routine analysis of various P. notoginseng pesticide residues. The pesticide residues in the main root samples of P. notoginseng were at a safe level and unlikely pose health risks to consumers.

Sodium chloride stimulates the biomass and astaxanthin production by Haematococcus pluvialis via a two-stage cultivation strategy.

A major challenge facing by astaxanthin industrialization is the low productivity and high production costs. This study established a two-stage cultivation strategy based on the application of NaCl to improve the production of biomass and astaxanthin by Haematococcus pluvialis. During the first growth stage, 12.5 mg L-1 NaCl led to a remarkable enhancement in biomass, which was 1.28 times compared with the control. Moreover, 2 g L-1 NaCl stimulated the astaxanthin content from 12.18 mg g-1 to 25.92 mg g-1 during the second induction stage. Simultaneously, salinity stress application increased the lipids and GABA contents, as well as the levels of Ca2+ and carotenogenic genes' expression, but suppressed the contents of carbohydrate and protein and high-light induced-ROS. This study proposed a simple and convenient strategy for efficient coproduction of biomass and astaxanthin and provides insights into the underlying mechanism of astaxanthin biosynthesis in H. pluvialis induced by salinity stress.

Coupling of myo-inositol with salinity regulates ethylene-induced microalgal lipid hyperproduction in molasses wastewater.

With the goal of cost-effective and high-efficient microalgae-based biodiesel production, this study evaluated the feasibility of the joint strategy concerning myo-inositol (MI) and salinity stress on lipid productivity of Monoraphidium sp. QLY-1 in molasses wastewater (MW). The maximal lipid productivity (147.79 mg L-1 d-1) was obtained under combined 0.5 g L-1 MI and 10 g L-1 NaCl treatment, which was 1.40-fold higher than the control. Meanwhile, the nutrients removal from MW was markedly increased under MI-NaCl treatment. Moreover, exogenous MI upregulated key lipogenic genes' expressions, activated autophagic activity and ethylene (ET) signaling, and ultimately alleviated the salinity-induced damage via reactive oxygen species (ROS) signaling. Further pharmacologic experiment confirmed the indispensable role of ET in the lipogenesis progress under the combined treatment. These data demonstrated the combined salinity stress and MI treatment to be capable for lipid hyperproduction and wastewater nutrients removal, which contributes to practically integrating the microalgae cultivation with wastewater treatment.

A joint strategy comprising melatonin and 3-methyladenine to concurrently stimulate biomass and astaxanthin hyperaccumulation by Haematococcus pluvialis.

Haematococcus pluvialis is a commercial microalgae used for natural astaxanthin production. This study aims to investigate the roles of melatonin (MT) and 3-methyladenine (3-MA) in regulating the cell growth and biosynthesis of astaxanthin and fatty acids under adverse conditions by H. pluvialis. Upon the dual treatments, the maximum astaxanthin concentration (46.78 mg L-1) was 2.39- and 1.35-fold higher compared with the control and MT treatments, respectively. Concomitantly, the combined application of MT and 3-MA suppressed autophagy but promoted the production of biomass and lipids and upregulated carotenogenesis, lipogenesis and antioxidant enzyme-related genes at the transcriptional level, which were linked with astaxanthin and lipid biosynthesis and oxidative stress. Additionally, astaxanthin exhibited a noticeable increase under MT coupled with 3-MA in the other two strains of H. pluvialis. This study proposed a potential method for astaxanthin induction and provided insights into the function of autophagy in modulating cell growth and astaxanthin synthesis.

Melatonin and calcium act synergistically to enhance the coproduction of astaxanthin and lipids in Haematococcus pluvialis under nitrogen deficiency and high light conditions.

This study focused on the influence of integrating melatonin (MT) and calcium (Ca2+) on the simultaneous accumulation of astaxanthin and lipids in Haematococcus pluvialis under abiotic stress conditions. Compared with the control condition, MT induction enhanced astaxanthin and lipid contents by 65.89% and 27.38%, respectively. The highest contents of astaxanthin and lipids under combined exposure to MT and Ca2+ were 3.8% and 49.53%, respectively, which were 1.13- and 1.21-fold higher than those of cells treated with MT alone. The application of MT and Ca2+ also promoted the expression of carotenogenic and lipogenic genes and increased the levels of Ca2+ and γ-aminobutyric acid (GABA) but decreased reactive oxygen species (ROS) levels. Further evidence indicated that the increased cellular Ca2+ could promote astaxanthin biosynthesis under MT induction by regulating carotenogenic gene levels and GABA and ROS signalling. The integrated strategy efficiently improved the coproduction of astaxanthin and lipids in H. pluvialis.

Melatonin attenuates nicotine-induced autophagy and neurological changes by decreasing the production of reactive oxygen species.

Purpose: The aim of this study was to explore the mechanism of neurological changes underlying the toxicity of nicotine.Materials and methods: Rat pheochromocytoma 12 (PC12) cells and human neuroglia (HM) cells were used. The ROS levels of the cells were detected by the FACScan. Autophagy flux was monitored by a tandem monomeric RFP-GFP-tagged LC3 lentivirus. The autophagic proteins LC3, SQSTM1/p62 and Beclin1 were detected by western blot assay. In order to evaluate the effects of nicotine and melatonin on the morphological changes of neurons, primary cortical neurons were obtained and immunocytochemistry of TUBB3 tubulin were conducted.Results: Nicotine increased the levels of reactive oxygen species (ROS) in PC12 and HM cells in a concentration-dependent manner. Microscopy showed increased autophagic flux in nicotine-treated PC12 cells. Subsequent western blotting results showed that nicotine induced increase in the levels of LC3B-II and Beclin1, and decreased SQSTM1/p62 in a concentration-dependent manner. Finally, nicotine treatment reduced the length of TUBB3-positive axons and dendrites. Melatonin, a mitochondrially targeted antioxidant, reduced the ROS level, and blocked autophagy activation and the morphologic structural changes induced by nicotine.Conclusions: Our results suggested that the role of nicotine in neuronal toxicity maybe through the induction of ROS and the subsequent activation of autophagy. These effects could be restored by melatonin.

γ-Aminobutyric acid (GABA) regulates lipid production and cadmium uptake by Monoraphidium sp. QLY-1 under cadmium stress.

This study explored the effects of γ-aminobutyric acid (GABA) on the production of biomass and lipids and on the uptake of Cd2+ by microalgae under cadmium (Cd) stress. Compared with the control and Cd stress alone, 2.5 mM GABA increased the maximum lipid content (55.37%) by 49.37% and 9.42%, respectively. GABA application resulted in increased contents of protein and glutathione (GSH) and in upregulated activity of α-amylase but decreased contents of starch, reactive oxygen species (ROS) and Cd2+, with no effect on subsequent biodiesel quality. Additional analysis of GABA further indicated that increased cellular GABA contents could promote lipid synthesis and reduce Cd accumulation by regulating the expression levels of lipogenesis genes, ROS signalling and mineral nutrient uptake under Cd stress. Collectively, these findings show that GABA not only increases lipid production in microalgae but also is involved in the mechanisms by which microalgae respond to Cd stress.

[Determination of 153 pesticide residues in tea using on-line gel permeation chromatography-gas chromatography/mass spectrometry].

A method was developed for the determination of 153 pesticide residues in tea using on-line gel permeation chromatography-gas chromatography/mass spectrometry (GPC-GC/ MS). The pesticide residues were extracted with acetonitrile under ultrasonic operation, and the extract was first cleaned up with an ENVI-carb solid phase extraction column and then separated and detected with the on-line GPC-GC/MS system. The recoveries of the method ranged from 73.32% to 117.05% with the relative standard deviations (RSDs) from 0.76% to 13.18%. The limits of detection and the limits of quantification were 0.0003-0.006 mg/kg and 0.001-0.02 mg/kg, respectively. This method is simple, rapid and characterized with acceptable sensitivity and accuracy to meet the requirements for the analysis of multiple pesticide residues in tea.