Study on the Structure and Bioactivity of Ganoderma lucidum Polysaccharides under Cassava Stalk Stress.

Various carbon sources affect the growth of the G. lucidum fruiting body, and the cassava stalk is considered a promising carbon source for G. lucidum. The composition, functional group characteristics, molecular weight distribution, antioxidant activity in vitro, and growth effect of L. rhamnosus LGG of G. lucidum polysaccharides (GLPs) under cassava stalk stress were investigated by gas chromatography-mass spectrometry, near-infrared spectroscopy, and gel chromatography. The results showed that GLPs consisted of D-glucose, D-galactose, and seven other monosaccharides. The end of the sugar chain had ß-D-Glc and ß-D-Gal configurations. The total sugar content in GLP1 was the highest (4.07%), and GLP1, GLP2, GLP3, and GLP5 had the ß-D-Gal configuration, while GLP4 and GLP6 had the ß-D-Glc configuration. The greater the proportion of cassava stalk, the greater the maximum molecular weight of GLPs. The total antioxidant capacities of GLPs obtained from different cassava stalks significantly varied, as well as their stimulating effects on the L. rhamnosus LGG growth. Higher concentrations of GLPs corresponded to the more intensive growth of L. rhamnosus LGG. This study provided essential data support for cassava stalk as a carbon source in G. lucidum cultivation.

Analysis of Lipids in Green Coffee by Ultra-Performance Liquid Chromatography-Time-of-Flight Tandem Mass Spectrometry.

Lipid components in green coffee were clarified to provide essential data support for green coffee processing. The types, components, and relative contents of lipids in green coffee were first analyzed by ultra-performance liquid chromatography-time-of-flight tandem mass spectrometry (UPLC-TOF-MS/MS). The results showed that the main fatty acids in green coffee were linoleic acid (43.39%), palmitic acid (36.57%), oleic acid (8.22%), and stearic acid (7.37%). Proportionally, the ratio of saturated fatty acids/unsaturated fatty acids/polyunsaturated fatty acids was close to 5.5:1:5.2. A total of 214 lipids were identified, including 15 sterols, 39 sphingosines, 12 free fatty acids, 127 glycerides, and 21 phospholipids. The main components of sterols, sphingosines, free fatty acids, glycerides, and phospholipids were acylhexosyl sitosterol, ceramide esterified omega-hydroxy fatty acid sphingosine, linoleic acid, and triglyceride, respectively. UPLC-TOF-MS/MS furnished high-quality and accurate information on TOF MS and TOF MS/MS spectra, providing a reliable analytical technology platform for analyzing lipid components in green coffee.

Anticancer effects of zapotin flavone in human gastric carcinoma cells are mediated via targeting m-TOR/PI3K/AKT signalling pathway.

The current study aims to investigate the anticancer effects of zapotin flavone in human gastric carcinoma cells. MTT assay was performed to determine the cytotoxicity effects of zapotin against the gastric cancer cells (SNU-1) and normal gastric cells (GES-1). SNU-1 cell morphology was analyzed through phase-contrast microscopy. Apoptosis was identified through DAPI staining assay and quantified through annexin V/propidium iodide (PI) staining. The effects on cell migration and invasion were carried out through transwell assay. Apoptosis and m-TOR/PI3K/AKT signalling pathway related proteins were analysed through western blotting. Proliferation rate of gastric cancer SNU-1 cell line declined with enhanced zapotin concentrations in comparison to normal GES-1 cells. Substantial morphological changes after zapotin exposure, including nuclear condensation and membrane rupture was observed. Further, increasing number of apoptotic cells, suppression of both cell migration and invasion was observed with increased zapotin concentrations. Finally, western blotting indicated significant blocking of m-TOR/PI3K/AKT signalling pathway. We conclude that zapotin can act as a potential drug against the gastric cancer.

ERO1α mediates endoplasmic reticulum stress-induced apoptosis via microRNA-101/EZH2 axis in colon cancer RKO and HT-29 cells.

Although colon cancer is a leading and typical gastrointestinal tumor, there is little published data on the underlying molecular mechanisms of endoplasmic reticulum (ER) stress. Here, we investigated the role of ERO1α and its impact on microRNA (miR)-101 expression and ER stress in colon cancer cells. Cell ER stress was established by treating RKO or HT-29 cells with 1 µM thapsigargin (THG). Cell biological behaviors were detected using CCK-8, bromodeoxyuridine assay, flow cytometry and western blot. We also investigated the expression of ERO1α and miR-101 after THG treatment using RT-qPCR. Moreover, effects of ERO1α and miR-101 on ER stress of colon cancer cells were detected. Additionally, miR-101 impact on EZH2 expression and relevance of this regulation was confirmed by RT-qPCR and luciferase reporter. The regulation of miR-101/EZH2 axis and Wnt/ß-catenin pathway in ER stress were investigated. Our results demonstrated that THG induced ER stress in colon cancer cells. Silencing ERO1α further promoted ER stress-induced cell apoptosis. ERO1α knockdown up-regulated miR-101 expression and promoted colon cancer cell apoptosis via regulating miR-101. Surprisingly, miR-101 negatively regulated EZH2 expression via miRNA-mRNA targeting. Moreover, ER stress promoted colon cancer cell apoptosis via regulating miR-101/EZH2 axis. Wnt/ß-catenin pathway was also involved in the regulation of ERO1α/miR-101/EZH2 in ER stress of colon cancer cells. These findings illustrated that silencing ERO1α regulated ER stress-induced apoptosis via miR-101/EZH2 axis in RKO and HT-29 cells.

LncRNA MEG3 promotes endoplasmic reticulum stress and suppresses proliferation and invasion of colorectal carcinoma cells through the MEG3/miR-103a-3p/PDHB ceRNA pathway.

LncRNA maternally expressed gene 3 (MEG3) is a potential prognostic and diagnostic biomarker in colorectal carcinoma (CC). However, its cellular functions and mechanism remain not fully uncovered. Relative expression of MEG3, miRNA (miR)-103a-3p, and pyruvate dehydrogenase E1 subunit beta (PDHB) was detected by RT-qPCR and western blotting. Cell proliferation was measured by CCK-8 assay, colony formation assay, and flow cytometry, as well as xenograft tumor assay. Transwell assay examined cell invasion. Endoplasmic reticulum (ER) stress was evaluated by western blotting. Dual-luciferase reporter assay and RNA immunoprecipitation determined the relationship between miR-103a-3p and MEG3 or PDHB. Expression of MEG3 was downregulated in human CC tumor tissues and cells (SW620 and HCT116), accompanied by higher miR-103a-3p and lower PDHB. Restoring MEG3 suppressed cell viability, colony formation ability, and invasion, arrested cell cycle, and induced apoptosis rate in SW620 and HCT116 cells, as well as promoted expression of ER stress-related proteins (GRP78, ATF6, CHOP, caspase-3, and caspase-9). Furthermore, MEG3 overexpression hindered tumor growth and facilitated ER stress in vivo. Molecularly, miR-103a-3p was a target of MEG3, and further targeted PDHB. Similarly, in function, blocking miR-103a-3p suppressed CC in vitro by affecting proliferation, invasion, and ER stress; in addition, restoring miR-103a-3p partially counteracted the suppressive role of MEG3 in CC cells. MEG3 sponged miR-103a-3p to suppress CC malignancy by inducing ER stress and inhibiting cell proliferation and invasion via upregulating PDHB, suggesting a novel MEG3/miR-103a-3p/PDHB ceRNA pathway.

Factors related to spiritual health in Chinese haemodialysis patients: A multicentre cross-sectional study.

Aim: This study aimed to investigate the current situation of the spiritual health of maintenance haemodialysis (MHD) patients in China and analyse the influencing factors. Methods: A total of 418 patients who underwent maintenance haemodialysis in three grade A tertiary hospitals were selected. The influencing factors were evaluated with demographic questionnaire, the Functional Assessment of Chronic Illness Therapy-Spiritual Well-Being (FACIT-Sp-12), Family APGAR Index, Herth Hope Index (HHI) and Acceptance of Illness Scale (AIS). Results: Spiritual health was positively correlated with the HHI, Family APGAR and AIS scores. Nationality, HHI score, Family APGAR score and AIS score were independent influencing factors of spiritual health. MHD patients had a moderate level of spiritual health. Nationality, hope, family function and acceptance of illness were significant predictors of spiritual health. Patients who have higher hope levels, better family functioning and better illness acceptance may maintain better spiritual health.

Serum starvation induces cell death in NSCLC via miR-224.

Purpose: Increasing evidence suggests that microRNAs (miRNAs) may be involved in the occurrence and progression of non-small cell lung cancer (NSCLC). In the present study, we used serum-starved A549 cells emulating tumor under a nutrient depletion stress in the microenvironment. Patients and methods: We first detected the expression level of miR-224 between tumor tissues and the adjacent normal tissues. We analyzed the expression levels of miR-224 and its predicted target phosphatase and tensin homolog (PTEN) using quantitative real-time PCR (qRT-PCR) in starved A549 cells. Following transfection with miR-224 mimic or inhibitor in starved A549 cells, MTT assay, Annexin V FITC/PI staining, and LC-3 immunofluorence staining were performed to investigate the roles of miR-224 on proliferation, apoptosis, and autophagy. Next, the expression of apoptosis-related protein Bax and Bcl-2, autophagy-related proteins LC3, PI3K signaling, and target PTEN were measured using qRT-PCR and Western blot assays. The direct interaction between miR-224 and PTEN was validated with a dual luciferase assay. Results: We found that the expression level of miR-224 in tumor tissues was significantly higher when compared with the adjacent normal tissues. We discovered a reciprocal expression pattern between miR-224 and PTEN in starved A549 cells, and transfection with miR-224 mimic led to down-regulation of PTEN. A dual luciferase assay further confirmed the direct interaction between miR-224 and 3'UTR of PTEN. Transfection with miR-224 mimic in starved A549 cells resulted in enhanced cell proliferation, reduced apoptosis, and autophagy, accompanied by increased expression of anti-apoptotic protein Bcl-2, decreased expression of pro-apoptotic protein Bax, and autophagy-related protein LC3. Activation of PI3K was observed in miR-224 mimic transfected cells. The reverse effects by the miR-224 inhibitor in all experiments were observed. Conclusion: Taken together, we proved that miR-224 might play essential roles in cellular functions of nutrient-depleted A549 cells possibly through regulating the target PTEN and downstream signal PI3K, suggesting the potential of miR-224 to be a therapeutic target for NSCLC therapy.

Epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer (NSCLC) of Yunnan in southwestern China.

To investigate the Epidermal Growth Factor Receptor (EGFR) mutation status in non-small cell lung cancer (NSCLC) in Yunnan province in southwestern China, we detected EGFR mutation by Amplification Refractory Mutation System (ARMS) polymerase chain reaction (PCR) using DNA samples from 447 pathologically confirmed NSCLC specimens (175 tissue, 256 plasma and 16 cytologic samples). The relationship between EGFR mutations and demographic and clinical factors were further explored. Subgroup analyses according to sample type (tissue and plasma) and histological type (adenocarcinoma) were done. We found the mutation rate was 34.9% in overall patients (42.3%, 29.7%, and 37.5% for tissue, plasma, and cytologic samples respectively). We found female (p < 0.0001), no smoking (p = 0.001), adenocarcinoma (p < 0.0001), and tissue specimen (p = 0.026) were associated with higher EGFR mutation rate. The most common mutations were exon 19 deletions (40%) and L858R point (30%) mutation. Interestingly, NSCLC patients from Xuanwei harbored a strikingly divergent mutational pattern for EGFR when compared with non-Xuanwei patients (higher G719X, G719X+S768I mutations, but lower 19 deletion and L858R mutations). Generally, EGFR mutation rate and pattern in Yunnan province was in accord with other Asian populations. However, Xuanwei subgroup showed strikingly divergent EGFR mutation spectrum from other general population. Our analysis also indicated that cftDNA analysis for EGFR mutations detection was feasibility for the patients lacking sufficient tissue for molecular analyses.