Silencing FUT4 Inhibits the Progression of Osteosarcoma through Activation of FOXO1.

BACKGROUND: It has been reported that inhibition of Fucosyltransferase4 (FUT4) to activate Forkhead box O1 (FOXO1) can lead to apoptosis of cancer cells, however, the mechanism in osteosarcoma is still unclear. OBJECTIVE: To explore the biological significance of the connection between FUT4 and FOXO1 in osteosarcoma growth. METHODS: In vitro tests were conducted using the human osteoblast cell line and the osteosarcoma cell lines. QRT-PCR assay as well as western blot assay were used to ascertain the relative expression levels of FUT4 and FOXO1 in the cells. By using the CCK-8 assay, colony assay, EDU assay, wound healing assay and Transwell assay, osteosarcoma cells' ability to proliferate, migrate and invade were examined in relation to si- FUT4. TUNEL test was used to evaluate Si-impact FUT4's on KHOS and U2OS apoptosis in osteosarcoma cells. Western blot assay was used to identify the expression of proliferative, migrating and apoptosis-related protein markers in osteosarcoma cells KHOS and U2OS and the expression of important proteins in the Wnt/ ß-catenin signaling pathway. RESULTS: In comparison with osteoblasts, osteosarcoma cells expressed more FUT4. The osteosarcoma cells' capacities to proliferate, invade, and migrate were markedly inhibited by the inhibition of FUT4 expression, which also increased osteosarcoma cell apoptosis. The Wnt/ß-catenin signaling pathway was blocked by upregulating FOXO1 expression, which was in turn inhibited by inhibiting FUT4 expression. CONCLUSION: Osteosarcoma cells express more FUT4. The Wnt/ß-catenin signaling pathway has a significant effect on osteosarcoma cell death, and inhibition of FUT4 expression may target FOXO1 activation to decrease osteosarcoma cells' ability to proliferate, invade, and migrate.

Life cycle assessment for evaluation of novel solvents and technologies: A case study of flavonoids extraction from Ginkgo biloba leaves.

Innovative solvents such as deep eutectic solvents (DESs) and process intensification technologies assisted by ultrasound have been demonstrated to be promising pathways for enhancing solid-liquid extraction. Nevertheless, quantitative and systematic knowledge of their environmental impact is still limited. In this work, a case study of flavonoids extraction from Ginkgo biloba leaves was evaluated by using life cycle assessment (LCA) for comparison of three extraction scenarios. The first used DES as extractant (DESE), and the other two adopted ethanol, including heat reflux extraction (HRE), and ultrasound-assisted extraction (UAE). Among eight key midpoints investigated, all these from UAE were 10.0 %-80.0 % lower than from DESE and HRE except water consumption. The UAE was the eco-friendliest option due to its higher extraction yield, shorter duration and lower solvent consumption. The DESE exhibited the lowest water consumption, the highest freshwater ecotoxicity and human carcinogenic toxicity, while HRE had the highest impacts for the other 6 midpoints. Moreover, solvent production was the key contributor for all the categories. The standardized sensitivity analysis showed that the overall environmental footprint can be further decreased by 15.4 % for DESE pathways via substituting choline chloride/glycerine with choline chloride/ethylene glycol. Furthermore, all pathways using DESs had higher standardized impacts than those employing ethanol from sugarcane or wood. Replacing ethanol from maize with other feedstocks can significantly lessen the overall impacts, among which the UAE using ethanol from sugarcane demonstrated the least environmental impacts. The promotion of DESs as "green and sustainable" alternative to traditional solvents requires careful consideration.

Improved ergonomic layout design of metro control center based on virtual simulation technology and genetic algorithm.

For the existing control center layout methods do not have targeted measures to implement human factors requirements, this paper puts forward a specific human factors design method for metro control center layout. This method covers the overall process of ergonomic standards and actual engineering requirements in personal space determination, overall location design, and personnel post layout. It introduces discrete personnel simulation to assist in physical control center layout design for the controllers' safety. To make the posts between the unified lines and the connections between the lines more reasonable, an improved genetic algorithm is used to solve the Systematic Layout Planning. This design flow and algorithm can be quickly adapted according to the project's actual needs and the application of human factors guidelines. This method is also suitable for the layout planning and design of similar large spaces, large numbers of people, and complex associated control centers.

Iridoid-glycoside isolation and purification from Premna fulva leaves.

Premna fulva Craib, rich in iridoid glycosides, is widely used to treat periarthritis, osteoproliferation, pain, and other diseases. However, no studies have reported effective purification methods for obtaining iridoid glycosides as active materials. This paper describes an efficient strategy for separating iridoid glycosides from Premna fulva leaves using high-speed counter-current chromatography and preparative high-performance liquid chromatography. A two-phase solvent system, ethyl acetate/n-butanol/water (7.5:2.5:10, v/v), was selected for high-speed counter-current chromatography separation. The proposed method effectively separated and purified four iridoid glycosides and four lignans, including three new iridoid glycosides (4-6) and five known compounds (1-3, 7, 8), from Premna fulva leaves, indicating that high-speed counter-current chromatography combined with prep-HPLC can efficiently isolate catalpol derivatives from the genus Premna. Additionally, the in vitro anti-inflammatory activities of all isolated compounds were analyzed using lipopolysaccharide-stimulated RAW 264.7 cells, and the results indicated that six compounds (1 and 3-7) exhibited potential anti-inflammatory activities.

MiR-125b-5p/STAT3 Axis Regulates Drug Resistance in Osteosarcoma Cells by Acting on ABC Transporters.

Background: The poor prognosis of the highly malignant tumor osteosarcoma stems from its drug resistance and therefore exploring its resistance mechanisms will help us identify more effective treatment options. However, the effects of miR-125b-5p on drug resistance in osteosarcoma cells are still unclear. Methods: To study the effects of miR-125b-5p on drug resistance in osteosarcoma cells. Osteosarcoma-resistant miR-125b-5p was obtained from the databases GeneCards and g:Profiler. CCK8, western blot, and transwell were applied for the detection of the miR-125b-5p effects on proliferation, migration, invasion, apoptosis, and drug resistance in osteosarcoma. Bioinformatics is aimed at demonstrating the targeting factor miR-125b-5p, performing protein interaction enrichment analysis by Metascape, and finally validating by binding sites. Results: Upregulation of miR-125b-5p restrains proliferation, migration, and invasion of osteosarcoma and promotes apoptosis. In addition, miR-125b-5p can restore drug sensitivity in drug-resistant osteosarcoma. miR-125-5p restrains the signal transducer and inhibits the transcription 3 (STAT3) expression activator via targeting its 3'-UTR. STAT3 affects drug-resistant osteosarcoma to regulate the ABC transporter. Conclusion: miR-125b-5p/STAT3 axis mediates the drug resistance of osteosarcoma by acting on ABC transporter.

GUESS - A simple approach to accelerate optimization countercurrent separation.

The generally useful estimate of solvent systems (GUESS) method, which is based on thin layer chromatography, is a simple and practical method for selecting solvent systems for countercurrent chromatography (CCC). However, it is rarely used for complex samples derived from natural products. In this study, GUESS was used for CCC solvent system selection and polarity-adjusted CCC separations of several fractions, which were obtained from a silica gel column containing complex compositions with a broad polarity from Salvia bowleyana Dunn. The GUESS method was performed on five fractions based on solvent systems in the n-hexane-ethyl acetate-methanol-water (HEMWat) family. Based on the GUESS results, the optimal solvent systems were selected for CCC separation. Twelve diterpenoids were obtained from the five silica gel column fractions of S. bowleyana Dunn using elution-extrusion countercurrent chromatography (EECCC). These demonstrate that GUESS guidance and the polarity adjustment of the solvent system accelerate the optimization of CCC separation conditions and simplify the process of accommodating a broad polarity of components in complicated mixture fractions. We therefore confirmed the feasibility and advantage of the GUESS method for complex natural chemical component separations.

The generally useful estimate of solvent systems method facilitates off-line two-dimensional countercurrent chromatography for isolating compositions from Siraitia grosvenorii roots.

Solvent system selection is a crucial and the most time-consuming step for successful countercurrent chromatography separation. A thin-layer chromatography-based generally useful estimate of solvent systems method has been developed to simplify the solvent system selection. We herein utilized the method to select a solvent system for off-line two-dimensional countercurrent chromatography to separate chemical compositions from a complex fraction of the Siraitia grosvenorii root extract. The first-dimensional countercurrent separation using chloroform/methanol/water (10:5.5:4.5, v/v/v) yielded four compounds with high purity and three mixture fractions (Fr I, III, and VII). The second-dimensional countercurrent separation conducted on Fr I, III, and VII using the hexane/ethyl acetate/methanol/water (4:6:6:4, 3:7:3:7, v/v/v) and chloroform/methanol/water (10:9:6, v/v/v) solvent systems, respectively, produced another four compounds. Four triterpenoids and four lignans were finally isolated, including two novel compounds. Hence, the generally useful estimate of solvent systems method is a feasible and efficient approach for selecting an applicable solvent system for separating complex samples. In addition, the off-line two-dimensional countercurrent chromatography method can improve both the peak resolution and the capacity of countercurrent chromatography.

The Effect of Task Interruption on Working Memory Performance.

OBJECTIVE: This study used electroencephalography to explore the behavioral and electrophysiological effects of task interruption on performance. BACKGROUND: Task interruption is known to harm work performance, especially on working memory-related tasks. However, most studies pay little attention to cognitive processes by exploring brain activity and ignore the cumulative effect of sequential interruptions. METHOD: Thirty-four healthy participants performed a spatial 2-back in three conditions: (1) interruptions with simple math questions, (2) suspensions with prolonged fixation cross, and (3) a pure 2-back. The measured outcomes comprise performance data, ERP amplitudes, EEG power, and subjective workload. RESULTS: Work performance decreased in the resumption trials, and cumulative interruptions had a more destructive effect on performance. EEG results showed that the P2 and P3 amplitudes induced by the 2-back task significantly increased after interruptions; theta and alpha power increased after interruptions. The P3 amplitude and alpha power induced by interruptions were significantly higher than that induced by suspensions. CONCLUSION: Behavioral data revealed the disruptive effect of interruptions on postinterruption performance and the cumulative effect of interruptions on accuracy. Changes in ERP amplitudes and EEG power indicate the mechanisms of attention reallocation and working memory during interruptions. Larger P3 amplitudes and alpha power after interruptions than after suspensions suggested the inhibition of irrelevant information. These results may support the memory for goals model and improve the understanding of the effects of interruption on working memory. APPLICATION: Focusing upon the mechanisms at play during the interruption process can support interruption management to ensure work safety and efficiency.

The moderation effects of task attributes and mental fatigue on post-interruption task performance in a concurrent multitasking environment.

In a concurrent multitasking environment, performing many types of tasks increases task complexity, and working long hours makes a person susceptible to mental fatigue. Emerging technologies may lead to more task interruptions. This study examines the effects of task attributes and mental fatigue on interrupted task performance in a concurrent multitasking environment. Thirty-four participants performed the MATB-Ⅱ under eight conditions (two-level task interruption, two-level task complexity, two-level fatigue). The results revealed the significant interaction effects of interruption × task complexity and of interruption × fatigue state. The findings show that more time is required to return to a complex primary task, and there are differences among subtask types. Mental fatigue negatively affects primary task performance, workload, and the resumption lag after an interruption. The findings are explained by the increasing information cues needed to resume complex tasks and the negative effect of fatigue on memory activation.

Preparative isolation of diterpenoids from Salvia bowleyana Dunn roots by high-speed countercurrent chromatography combined with high-performance liquid chromatography.

The root of Salvia bowleyana Dunn (Lamiaceae) is used as a traditional Chinese medicine that has multiple therapeutic effects. In this study, an efficient strategy was developed to separate diterpenoid compounds, which are the main active ingredients in Salvia bowleyana Dunn roots, from complex crude extracts by high-speed countercurrent chromatography combined with preparative high-performance liquid chromatography. A two-phase solvent system comprising n-hexane-ethyl acetate-methanol-water (7:3:7:3, v/v/v/v) was selected for high-speed countercurrent chromatographic separation. Three major diterpenoids, 6α-hydroxysugiol (7), sugiol (8), and 6, 12-dihydroxyabieta-5,8,11,13-tetraen-7-one (9) were obtained at purities of 98.9, 95.4, and 96.2%, respectively, and minor diterpenoids were enriched via one-step separation. The enriched minor diterpenoids were further purified by continuous preparative high-performance liquid chromatography to yield two new norabietanoids (1, 6) and four known compounds (2-5). The structures of these new compounds were determined using NMR spectroscopy, high-resolution electrospray ionization mass spectrometry, and electronic circular dichroism spectroscopy. The results suggest that high-speed countercurrent chromatography combined with preparative high-performance liquid chromatography efficiently isolates diterpenoids, including minor components, from complex natural products.

Fatigue-Related Effects in the Process of Task Interruption on Working Memory.

Interruption generally has a negative effect on performance by affecting working memory (WM). However, the neural mechanism of interruption has yet to be understood clearly, and previous studies have largely ignored the role of fatigue state. To address these issues, the present study explores the behavioral and electrophysiological effects of interruption on WM performance using electroencephalography (EEG) data. The moderating effect of fatigue is also explored. The participants performed spatial 2-back tasks with math task interruption, suspension interruption, and non-interruption under different fatigue states. The results show that interruption led to increased alpha activity and P300 amplitude, indicating inhibitory control to interference from irrelevant information. Analysis of P200 amplitude revealed that interruption affected attentional reallocation when resuming the primary task. Increased theta power indicated an increased demand for information maintenance during the interruption. A speeding-up effect was discovered after interruption; however, fatigue impaired cognitive ability and further exacerbated the negative effects of interruption on WM and behavioral performance. These findings contribute to a better understanding of cognitive activity during the interruption and of the interaction with fatigue, and provide further support for the theory of memory for goals (MFG).

Vicenin 3 ameliorates ECM degradation by regulating the MAPK pathway in SW1353 chondrocytes.

Aberrant destruction of the articular extracellular matrix (ECM) has been considered to be one of the pathological features of osteoarthritis (OA) which results in chondrocyte changes and articular cartilage degeneration. The MAPK signaling pathway serves a key role by releasing cartilage-degrading enzymes from OA chondrocytes. However, the use of MAPK inhibitors for OA is hindered by their potential long-term toxicity. Vicenin 3 is one of the major components of the Jian-Gu injection which is effective in the clinical treatment of OA. However, its potential impact on OA remain poorly understood. Therefore, the present study aimed to assess the effects of vicenin 3 on interleukin (IL)-1ß-treated SW1353 chondrocytes, which mimic the microenvironment of OA. These chondrocytes were pretreated with vicenin 3 (0, 5 and 20 µM) for 1 h and subsequently stimulated with IL-1ß (10 ng/ml) for 24 h. Nitric oxide (NO) production was measured using the Griess reaction, whereas the production of prostaglandin E2 (PGE2), matrix metalloproteinases (MMPs), A disintegrin-like and metalloproteinase with thrombospondin motifs (ADAMTSs), collagen type II and aggrecan were measured using ELISA. The mRNA expression of MMPs and ADAMTSs were measured using reverse transcription-quantitative PCR. The protein expression levels of MAPK were measured using western blotting. Vicenin 3 was found to significantly inhibit IL-1ß-induced production of NO and PGE. Increments in the expression levels of MMP-1, MMP-3, MMP-13, ADAMTS-4 and ADAMTS-5 induced by IL-1ß, in addition to the IL-1ß-induced degradation of collagen type II and aggrecan, were all reversed by vicenin 3 treatment. Furthermore, vicenin 3 suppressed IL-1ß-stimulated MAPK activation, an effect that was similar to that exerted by SB203580, a well-known p38 MAPK inhibitor. In conclusion, vicenin 3 may confer therapeutic potential similar to that of the p38 MAPK inhibitor for the treatment of OA.

Hypoglycemic effects of esculeoside A are mediated via activation of AMPK and upregulation of IRS-1.

BACKGROUND: Tomato fruit (Lycopersicon esculentum Mill.) has been suggested to be useful for the prevention of diabetes. Esculeoside A is the main saponin compounds in tomatoes. This study investigated the hypoglycemic effects and the underlying mechanism of esculeoside A in C57BLKS/Leprdb (db/db) mice. METHODS: Wild-type C57BLKS (db/dm) mice were used in the db/dm mouse group and db/db mice were randomly divided into 2 groups: untreated and treated db/db mouse groups. Esculeoside A (100 mg/kg) was administered by gavage for 56 days to the treated db/db mouse group. Distilled water was administered to the db/dm mouse group and the untreated db/db mouse group. The blood and liver biochemical parameters and the expression of liver insulin signaling-related proteins were examined. RESULTS: The results showed that esculeoside A reduced the fasting blood glucose (FBG) levels and improved the glucose tolerance. Further investigation revealed that hepatic protein expressions of total AMP-activated protein kinase (T-AMPK), phosphorylated AMP-activated protein kinase (p-AMPK), insulin receptor substrate-1 (IRS-1), and glucokinase (GCK) were significantly upregulated after esculeoside A treatment. In contrast, the hepatic protein expression of phosphoenolpyruvate carboxykinase (PEPCK) was significantly downregulated by esculeoside A treatment. CONCLUSION: These findings suggested that esculeoside A has a potential of alleviating the metabolic abnormalities in db/db mice via regulation of AMPK/IRS-1 pathway. Our findings supported a possible application of esculeoside A as a functional supplement for diabetes treatment.

Preparation of flavone di-C-glycoside isomers from Jian-Gu injection (Premna fulva Craib.) using recycling counter-current chromatography.

Jian-Gu Injection (JGI, Premna fulva Craib) has been demonstrated to be effective in the clinical treatment of bone hyperplasia. However, an effective purification method of the JGI flavone C-glycosides as reference materials is not available. The present work developed a recycling counter-current chromatography approach to prepare these materials in high quality. An optimized biphasic solvent system composed of ethyl acetate/n-butanol/water (1:9:10, v/v/v) was employed to purify the five congeneric flavone C-glycosides, identified as apigenin 6,8-di-C-ß-d-glucopyranoside (vicenin 2), apigenin 6-C-ß-d-xylopyranosyl-8-C-ß-d-glucopyranoside (vicenin 1), apigenin 6-C-ß-d-xylopyranosyl-8-C-ß-d-galactopyranoside, apigenin 6-C-ß-d-glucopyranosyl-8-C-ß-d-xylopyranoside (vicenin 3), and apigenin 6-C-ß-d-galactopyranosyl-8-C-ß-d-xylopyranoside, by means of UHPLC-PDA-ESI-IT-ToF-MSn and NMR spectroscopy.

Phenolic Compounds from the Leaves of Castanopsis fargesii.

In the course of a phytochemical and chemotaxonomical investigation of Castanopsis species (Fagaceae), three new phenolic compounds, (3R,1'S)-[1'-(6″-O-galloyl-ß-d-gluco-pyranosyl)oxyethyl]-3-hydroxy-dihydrofuran-2(3H)-one (1), (2R,3S)-2-[2'-(galloyl)oxyethyl]-dihydroxybutanoic acid (2), and (3S,4S)-3-hydroxymethyl-3,4-dihydro-5,6,7-trihydroxy-4-(4'-hydroxy-3'-methoxyphenyl)-1H-[2]-benzopyran-1-one (3) were isolated from the fresh leaves of Castanopsis fargesii. In addition, a known phenolic glycoside, gentisic acid 5-O-α-l-rhamnopyranosyl-(1→2)-ß-d-glucopyranoside (4) was also isolated and identified. Their structures were elucidated by means of spectroscopic methods including one- and two-dimensional NMR techniques.

Studies on chemical fingerprints of Siraitia grosvenorii fruits (Luo Han Guo) by HPLC.

A novel, efficient, and accurate fingerprinting method using high performance liquid chromatography-photodiode array detection has been developed and optimized for the investigation and demonstration of the variance in chemical properties among Siraitia grosvenorii fruits from different origins. The effects of growth stages, cultivated varieties, collection locations, and fruit portions of the herb on chromatographic fingerprints were examined. Eleven compounds were identified on chromatograms by comparing the retention time and UV spectrum of each peak separately with those of external references. The results revealed that chromatographic fingerprints, combining similarity or hierarchical clustering analysis along with reference compounds, could efficiently identify and distinguish S. grosvenorii fruits from different sources, which provided helpful clues for studying the plants' secondary metabolites and benefitted quality control.

New pregnane glycosides from Brucea javanica and their antifeedant activity.

Three new pregnane glycosides, 3-O-ß-D-glucopyranosyl-(1→2)-α-L-arabinopyranosyl-(20R)-pregn-5-ene-3ß,20-diol (1), 3-O-α-L-arabinopyranosyl-(20R)-pregn-5-ene-3ß,20-diol-20-O-ß-D-glucopyranoside (2), 3-O-α-L-arabinopyranosyl-(20R)-pregn-5-ene-3ß,20-diol-20-O-ß-D-glucopyranosyl-(1→2)-ß-D-glucopyranoside (3) were isolated along with four known compounds, 4-7, from the leaves and stems of Brucea javanica. Their structures were determined by detailed analyses of 1D- and 2D-NMR spectroscopic data. All of the compounds isolated from Brucea javanica were tested for the antifeedant activities against the larva of Pieris rapae. Compounds 1, 3, and 5 showed significant antifeedant activities after 72 h incubation.

[Determination of esculeoside A in Lycopersicon esculentum MILL by high performance liquid chromatography with evaporative light scattering detection].

A method for the determination of esculeoside A in Lycopersicon esculentum MILL by high performance liquid chromatography (HPLC) with evaporative light scattering detection (ELSD) was established. The results showed that esculeoside A had good linearity in the range of 0.61-3.05 mg with the correlation coefficient of 0. 999 5. The average recoveries were 97.9%-104.8% with the relative standard deviations (RSD) < or = 4.14% (n=5). The method is simple, sensitive and suitable for the analysis of esculeoside A and quality control for the raw material and extract of Lycopersicon esculentum MILL.