Growth performance, serum parameters, inflammatory responses, intestinal morphology and microbiota of weaned piglets fed 18% crude protein diets with different ratios of standardized ileal digestible isoleucine to lysine.

The present study was to explore the Ile requirement of piglets fed 18% crude protein (CP) diets. Two hundred and fifty 28-day-old Duroc × Landrace × Yorkshire piglets (8.37 ± 1.92 kg) were randomly divided into 5 dietary treatments (10 piglets per replicate, 5 barrows and 5 gilts per replicate) with 45%, 50%, 55%, 60%, 65% standardized ileal digestible (SID) Ile-to-Lys ratios, and the SID Lys was formulated to 1.19%. The experimental design consisted of two phases (d 1 to 14 and d 15 to 28). Results showed that average daily gain (ADG) had a tendency to quadratically increase as the SID Ile-to-Lys ratio increased (P = 0.09), and the optimum SID Ile-to-Lys ratios required to maximize ADG were 48.33% and 54.63% for broken-line linear model and quadratic polynomial model, respectively. Different SID Ile-to-Lys ratios had no significant effects on average daily feed intake and gain-to-feed ratio. Dry matter (P < 0.01), CP (P = 0.01), ether extract (P = 0.04), gross energy (P < 0.01) and organic matter (P < 0.01) digestibility increased quadratically. Serum total cholesterol levels decreased linearly (P = 0.01) and quadratically (P < 0.01); aspartate aminotransferase (P < 0.01), interleukin-1ß (P = 0.01), and tumor necrosis factor-α (P < 0.01) levels decreased quadratically; immunoglobulin G (P = 0.03) and immunoglobulin M (P = 0.01) concentrations increased quadratically. Serum Ser levels decreased linearly (P < 0.01) and quadratically (P = 0.01); Glu (P = 0.02), Arg (P = 0.05), and Thr (P = 0.03) levels decreased quadratically; Gly (P < 0.01) and Leu (P = 0.01) levels decreased linearly; Ile (P < 0.01) concentration increased linearly. Duodenal villus height (P < 0.01) and villus height to crypt depth ratio (P < 0.01) increased quadratically. The deficiency or excess of Ile decreased short chain fatty acid-producing bacteria abundance and increased pathogenic bacteria abundance. Overall, taking ADG as the effect index, the optimum SID Ile-to-Lys ratios of piglets offered 18% CP diets were 48.33% and 54.63% based on two different statistical models, respectively, and the deficiency or excess of lle negatively affected piglet growth rates and health status.

MUC1-EGFR crosstalk with IL-6 by activating NF-κB and MAPK pathways to regulate the stemness and paclitaxel-resistance of lung adenocarcinoma.

BACKGROUND: The chemotherapy resistance often leads to chemotherapy failure. This study aims to explore the molecular mechanism by which MUC1 regulates paclitaxel resistance in lung adenocarcinoma (LUAD), providing scientific basis for future target selection. METHODS: The bioinformatics method was used to analyse the mRNA and protein expression characteristics of MUC1 in LUAD. RT-qPCR and ELISA were used to detect the mRNA and protein expression, flow cytometry was used to detect CD133+ cells, and cell viability was detected by CCK-8 assay. The mRNA-seq was performed to analyse the changes in expression profile, GO and KEGG analysis were used to explore the potential biological functions. RESULTS: MUC1 is highly expressed in LUAD patients and is associated with a higher tumour infiltration. In paclitaxel resistance LUAD cells (A549/TAX cells), the expression of MUC1, EGFR/p-EGFR and IL-6 were higher than that of A549 cells, the proportion of CD133+ cells was significantly increased, and the expression of cancer stem cell (CSCs) transcription factors (NANOG, OCT4 and SOX2) were significantly up-regulated. After knocking down MUC1 in A549/Tax cells, the activity of A549/Tax cells was significantly decreased. Correspondingly, the expression of EGFR, IL-6, OCT4, NANOG, and SOX2 were significantly down-regulated. The mRNA-seq showed that knocking down MUC1 affected the gene expression, DEGs mainly enriched in NF-κB and MAPK signalling pathway. CONCLUSION: MUC1 was highly expressed in A549/TAX cells, and MUC1-EGFR crosstalk with IL-6 may be due to the activation of NF-κB and MAPK pathways, which promote the enrichment of CSCs and lead to paclitaxel resistance.

Multifunctional polylactic acid sensing fabric based on biomass flame retardants for intelligent fire early-warning.

Today, building materials emit many hazardous gases in the event of a fire, causing great harm to human health and the environment. Therefore, it is of great significance to develop bio-based flame retardant materials and to realize preventive measures to reduce fires or their damage. In this work, we fabricated a novel multifunctional fire early-warning polylactic acid-based fabric (MFR-PBF) by coating MXene nanosheet, phytic acid @ furfurylamine (PA@FA) and ammonium polyphosphate (APP) via an eco-friendly layer-by-layer assembly method. MFR-PBF showed outstanding flame retardancy including a limiting oxygen index value of 35 % and better char formation capacity. More importantly, MFR-PBF exhibited sensitive fire early-warning capability (∼1 s) and excellent cyclic alarm stability (>15 cycles) due to the excellent semiconductor responsiveness (light and heat) and the significant catalytic char formation effect. Moreover, MFR-PBF is comfortable, flexible and strong enough to sew onto firefighter uniform to detect a variety of human motions, which can be monitored in the internet by using a LoRa emitter and a gateway. In addition, the controllable heating performance rendered MFR-PBF as a potential portable heater. This work provides new insights into the preparation and application of intelligent fire early-warning fabrics in the smart fire protection and Internet of Things.

DNA methylation role in subgenome expression dominance of Juglans regia and its wild relative J. mandshurica.

Subgenome expression dominance plays a crucial role in the environmental adaptation of polyploids. However, the epigenetic molecular mechanism underlying this process has not been thoroughly investigated, particularly in perennial woody plants. Persian walnut (Juglans regia) and its wild relative, Manchurian walnut (Juglans mandshurica), are woody plants of great economic importance and are both paleopolyploids that have undergone whole-genome duplication events. In this study, we explored the characteristics of subgenome expression dominance in these 2 Juglans species and examined its epigenetic basis. We divided their genomes into dominant subgenome (DS) and submissive subgenome (SS) and found that the DS-specific genes might play critical roles in biotic stress response or pathogen defense. We comprehensively elucidated the characteristics of biased gene expression, asymmetric DNA methylation, transposable elements (TEs), and alternative splicing (AS) events of homoeologous gene pairs between subgenomes. The results showed that biased expression genes (BEGs) in 2 Juglans species were mainly related to external stimuli response, while non-BEGs were related to complexes that might be involved in signal transduction. DS genes had higher expression and more AS events while having less DNA methylation and TEs than homoeologous genes from the SS in the 2 Juglans species. Further studies showed that DNA methylation might contribute to the biased expression of gene pairs by modifying LTR/TIR/nonTIR TEs and improving the AS efficiency of corresponding precursor mRNAs in a particular context. Our study contributes to understanding the epigenetic basis of subgenome expression dominance and the environmental adaptation of perennial woody plants.

Survival analysis of palliative radiotherapy in patients with HER-2+ metastatic breast cancer.

Background: Whether radiotherapy can improve the long-term survival of HER-2+ metastatic breast cancer remains unclear. We launched this study to explore the effect of HER-2+ metastatic breast cancer patients through anti-HER-2 targeted therapy + radiotherapy. Methods: 488 HER-2 + metastatic breast cancer patients who received anti-HER2 targeted ± local radiotherapy from March 2006 to September 2021 were retrospectively collected. Patients were divided into a radiotherapy group (n=207) and a non-radiotherapy group (n=281) based on whether they received radiotherapy or not. 1: 1 propensity matching analysis was used to determine two groups of patients with similar baselines. Results: Before matching, the radiotherapy group (n=207) had a median overall survival (mOS) of 51.7 months (48.8-63.8), which was superior to the non-radiotherapy group's (n=281) mOS of 33.9 months (27.9-39.9) (P < 0.0001). Moreover, the radiotherapy group exhibited better 1-year (94.6% vs 83.9%), 3-year (70.8% vs 45.5%), and 5-year (43.3% vs 25.0%) survival rates compared to the control group. Propensity score matching analysis identified 135 pairs of baseline-matched patients. In the matched groups, the mOS was 57.2 (44.5-69.8) months in the radiotherapy group (n=135) and 34.1 (27.5-40.6) months in the non-radiotherapy group (n=135), showing a statistically significant difference (P < 0.0001). Additionally, the radiotherapy group demonstrated 1-, 3-, and 5-year survival rates of 93.2%, 71.5%, and 46.9%, respectively, while those in the non-radiotherapy group were 89.4%, 45.8%, and 22.2%, respectively. Multivariate Cox analysis revealed that the presence of brain metastasis, liver metastasis, and radiotherapy were identified as independent predictive factors significantly associated with OS. Conclusion: In patients with HER-2 positive metastatic breast cancer, radiotherapy was associated with better survival benefits compared to those who did not receive radiotherapy.

Dietary Forsythia suspensa extracts supplementation improves antioxidant status, anti-inflammatory functions, meat fatty acid deposition, and intestinal microbial community in finishing pigs.

This study aimed to determine the effects of Forsythia suspensa extracts (FSE) on performance, antioxidant status, inflammatory cytokines, meat quality, meat fatty acid composition, and gut microbial community in finishing pigs. Sixty-four pigs [Duroc × (Landrace × Yorkshire)] with an average initial body weight of 88.68 kg were randomly allotted to two dietary treatments, with eight replicate pens per treatment (four pens were barrows and four pens were gilts), four pigs per pen. The dietary treatments included a corn-soybean meal basal diet (CON) and an FS diet (basal diet + 100 mg/kg FSE; FS). Compared with CON, pigs fed FSE showed enhanced (P < 0.05) saturated fatty acid (SFA)/polyunsaturated fatty acid (PUFA) ratio, reduced (P < 0.05) lightness, and n-6/n-3 PUFA ratio, as well as tended to increase C20:5n3 content in the longissimus dorsi muscle. Moreover, pigs fed FSE showed decreased (P < 0.05) serum cortisol and tumor nuclear factor-α contents, and increased (P < 0.05) serum high-density lipoprotein cholesterol, superoxide dismutase, and glutathione peroxidase contents compared with CON. These pigs also tended to have increased serum total protein and immunoglobulin G contents, and decreased serum low-density lipoprotein cholesterol and interleukin-1ß contents compared with CON. In the colon, pigs fed FSE had a higher (P < 0.05) relative abundance of Bifidobacteriales at the order level, Lactobacillaceae and Bifidobacteriaceae at the family level, as well as Lactobacillus and Bifidobacterium at the genus level compared with CON. In conclusion, dietary Forsythia suspensa extract supplementation effectively improved antioxidant status and anti-inflammatory functions, as well as modulated meat fatty acid composition, and gut microbial community in finishing pigs.

Maternal Dietary Forsythia suspensa Extract Supplementation Induces Changes in Offspring Antioxidant Status, Inflammatory Responses, Intestinal Development, and Microbial Community of Sows.

This experiment aims to investigate the effect of maternal diet supplemented with Forsythia suspensa extract (FSE) on the performance, antioxidant status, inflammatory responses, intestinal development, and microbial community of sows. A total of 24 gestating sows (Landrace × Yorkshire) were assigned to 2 treatments with 12 sows per treatment. From d 107 of gestation to d 21 of lactation, sows were supplemented with a basal diet as control (CON) or an FSE diet (basal diet + 100 mg/kg FSE). Compared with CON, sows fed FSE showed lower (P < 0.05) wean-to-estrus interval, body weight loss, and higher (P < 0.05) average daily gain of suckling piglet. Sows fed FSE had reduced (P < 0.05) serum malondialdehyde (MDA) content and enhanced (P < 0.05) catalase and glutathione peroxidase (GSH-Px) contents at farrowing and weaning compared with CON. The suckling piglets of FSE-fed sows had increased (P < 0.05) mRNA expressions of nuclear factor erythroid-2 related factor 2, heme oxygenase-1 in the liver, and lower (P < 0.05) serum MDA content on d 0, 7, and 14 of lactation. Sows fed FSE had lower (P < 0.05) serum tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8) contents at farrowing and reduced (P < 0.05) serum IL-6 and IL-8 contents at weaning compared with CON. Piglets from FSE-fed sows had enhanced (P ≤ 0.05) villus height and villus height to crypt depth ratio in the jejunum, and higher (P < 0.05) protein expression of Occludin in jejunal mucosa compared with CON. Sows fed FSE tended to have higher (P = 0.09) relative abundance of Lactobacillus at genus level in feces at weaning compared with CON. Our results showed maternal diet supplemented with FSE in lactating sows could effectively induce improvement of performance, antioxidant status, anti-inflammatory function, intestinal morphology, barrier function, and microbial community.

Comprehensive Evolutionary Analysis of CPP Genes in Brassica napus L. and Its Two Diploid Progenitors Revealing the Potential Molecular Basis of Allopolyploid Adaptive Advantage Under Salt Stress.

Allopolyploids exist widely in nature and have strong environmental adaptability. The typical allopolyploid Brassica napus L. is a widely cultivated crop, but whether it is superior to its diploid progenitors in abiotic stress resistance and the key genes that may be involved are not fully understood. Cystein-rich polycomb-like protein (CPP) genes encode critical transcription factors involved in the response of abiotic stress, including salt stress. To explore the potential molecular basis of allopolyploid adaptation to salt stress, we comprehensively analyzed the characteristics and salt stress response of the CPP genes in B. napus and its two diploid progenitors in this study. We found some molecular basis that might be associated with the adaptability of B. napus, including the expansion of the CPP gene family, the acquisition of introns by some BnCPPs, and abundant cis-acting elements upstream of BnCPPs. We found two duplication modes (whole genome duplication and transposed duplication) might be the main reasons for the expansion of CPP gene family in B. napus during allopolyploidization. CPP gene expression levels and several physiological indexes were changed in B. napus and its diploid progenitors after salt stress, suggesting that CPP genes might play important roles in the response of salt stress. We found that some BnCPPs might undergo new functionalization or subfunctionalization, and some BnCPPs also show biased expression, which might contribute to the adaptation of B. napus under saline environment. Compared with diploid progenitors, B. napus showed stronger physiological responses, and BnCPP gene expression also showed higher changes after salt stress, indicating that the allopolyploid B. napus had an adaptive advantage under salt stress. This study could provide evidence for the adaptability of polyploid and provide important clues for the study of the molecular mechanism of salt stress resistance in B. napus.

Dietary live yeast supplementation alleviates transport-stress-impaired meat quality of broilers through maintaining muscle energy metabolism and antioxidant status.

BACKGROUND: This experiment was to investigate the effect of dietary live yeast (LY, 1 × 1010 CFU g-1 ) supplementation on serum metabolic parameters, meat quality as well as antioxidant enzyme activity of transported broilers. A total of 192 one-day-old broilers were randomly assigned to four treatments with six replicates and eight chicks per replicate: a basal diet without transportation (CON), a basal diet containing 0 (T), 500 (T + LY500 ) and 1000 mg kg-1 (T + LY1000 ) LY with 3 h of transportation after feeding for 42 days, respectively. The serum and muscle samples of broilers were collected immediately after 3 h of transportation. RESULTS: A higher (P < 0.05) final body weight and average daily weight gain were observed in T + LY1000 group compared with CON and T groups. The T + LY1000 group reduced (P < 0.05) the serum lactate contents and improved (P < 0.05) the pH24h and decreased (P < 0.05) the drip loss in muscles of transported-broilers. Also, the T + LY1000 group enhanced (P < 0.05) the total-antioxidant capacity and reduced (P < 0.05) the malondialdehyde in serum and muscles. Besides, the messenger RNA (mRNA) expression of avian uncoupling protein (avUCP) in muscles was down-regulated (P < 0.05) of T + LY1000 group compared with T group. CONCLUSION: Dietary LY supplementation alleviates transport-stress-impaired meat quality of broilers through maintaining muscle energy metabolism and antioxidant status. Therefore, LY may serve as a potential protector for broilers under transport stress in the future. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

CANT1 serves as a potential prognostic factor for lung adenocarcinoma and promotes cell proliferation and invasion in vitro.

BACKGROUND: Calcium-activated nucleotidase 1 (CANT1), functions as a calcium-dependent nucleotidase with a preference for UDP. However, the potential clinical value of CANT1 in lung adenocarcinoma (LA) has not been fully clarified. Thus, we sought to identify its potential biological function and mechanism through bioinformatics analysis and in vitro experiments in LA. METHODS: In the present study, we comprehensively investigated the prognostic role of CANT1 in LA patients through bioinformatics analysis and in vitro experiments. The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) were utilized to analyze the expression of CANT1 in LA patients and their clinical-prognostic value. The immunohistochemistry staining was obtained from the Human Protein Atlas (HPA). A Cox regression model was used to evaluate prognostic factors. Gene ontology (GO) and Gene set enrichment analysis (GSEA) was performed to explore the potential regulatory mechanism of CANT1 in the development of LA. Moreover, we also examined the relationship between CANT1 expression and DNA methylation. Finally, we did in vitro experiments to evaluate the biological behavior and role of CANT1 in LA cells (LACs). RESULTS: Our study showed that the CANT1 expression was significantly elevated in the LA tissues compared with the normal lung tissues. Increased CANT1 expression was significantly associated with the TN stage. A univariate Cox analysis indicated that high CANT1 expression levels were correlated with poor overall survival (OS) in LA. Besides, CANT1 expression was independently associated with OS in multivariate analysis. GO and GSEA analysis showed the enrichment of mitotic nuclear division, DNA methylation, and DNA damage. Then we found that the high expression of CANT1 is positively correlated with hypomethylation. The methylation level was associated with prognosis in LA patients. Finally, in vitro experiments indicated that knockdown of CANT1 resulted in decreased cell proliferation, invasion, and G1 phase cell-cycle arrest in LACs. CONCLUSION: The present study suggested that CANT1 may serve as a potential prognosis biomarker in patients with LA. High CANT1 expression and promoter demethylation was associated with worse outcome. Finally, in vitro experiments verified the biological functions and behaviors of CANT1 in LA.

Prognostic value of SEC61G in lung adenocarcinoma: a comprehensive study based on bioinformatics and in vitro validation.

BACKGROUND: Studies have shown that the Sec61 gamma subunit (SEC61G) is overexpressed in several tumors and could serve as a potential prognostic marker. However, the correlation between SEC61G and lung adenocarcinoma (LUAD) remains unclear. In the current study, we aimed to demonstrate the prognostic value and potential biological function of the SEC61G gene in LUAD. METHODS: Public datasets were used for SEC61G expression analyses. The prognostic value of SEC61G in LUAD was investigated using the Kaplan-Meier survival and Cox analyses. The correlation between the methylation level of SEC61G and its mRNA expression was evaluated via cBioPortal. Additionally, MethSurv was used to determine the prognostic value of the SEC61G methylation levels in LUAD. Functional enrichment analysis was conducted to explore the potential mechanism of SEC61G. Also, single sample GSEA (ssGSEA) and TIMER online tool were applied to identify the correlation between SEC61G and immune filtration. Furthermore, cell functional experiments were conducted to verify the biological behavior of SEC61G in lung adenocarcinoma cells (LAC). RESULTS: SEC61G was upregulated in pan-cancers, including LUAD. High SEC61G expression was significantly correlated with worse prognosis in LUAD patients. Multivariate analysis demonstrated that high SEC61G expression was an independent prognostic factor in the TCGA cohort. (HR = 1.760 95% CI: 1.297-2.388, p < 0.001). The methylation level of SEC61G negatively correlated with the SEC61G expression (R = - 0.290, p < 0.001), and patients with low SEC61G methylation had worse overall survival. (p = 0.0014). Proliferation-associated terms such as cell cycle and cell division were significantly enriched in GO and KEGG analysis. Vitro experiments demonstrated that knockdown of SEC61G resulted in decreased cell proliferation, invasion and facilitated apoptosis in LAC. GSEA analysis found that SEC61G expression was associated with the E2F targets. Moreover, SEC61G expression was negatively correlated with the immune cell infiltration including CD4+ T cell, CD8+ T cell, B cell, macrophage, neutrophil, and dendritic cell. CONCLUSION: Our study indicated that overexpression of SEC61G was significantly associated with poor prognosis of LUAD patients and the malignant phenotypes of LUAD cells, suggesting that it could be a novel prognostic biomarker and potential therapeutic target of LUAD.

Enzyme-Treated Soybean Meal Enhanced Performance via Improving Immune Response, Intestinal Morphology and Barrier Function of Nursery Pigs in Antibiotic Free Diets.

This study aims to investigate the effects of ESBM on performance, antioxidant status, immune response, and intestinal barrier function of nursery pigs in antibiotic free diets compared with EFS. A total of 32 Duroc × (Landrace × Yorkshire) barrows (initial body weight of 8.05 ± 0.66 kg, weaned on d 28) were selected and allocated to two treatments with 16 replicates per treatment and one pig per replicate using a complete random design. The treatments included an EFS group (basal diet + 24% EFS; EFS) and an ESBM group (basal diet + 15% ESBM; ESBM). Corn was used to balance energy and diets were iso-nitrogenous at about 18% crude protein. The experiment lasted for 14 days and pigs were slaughtered for sampling on d 14. Compared with EFS, pigs fed ESBM showed enhanced (p < 0.05) gain to feed ratio and average daily gain and a reduced (p < 0.05) diarrhea score. These pigs had increased (p < 0.05) contents of glutathione peroxidase, catalase, superoxide dismutase, IgG, interleukin-10, and ferric reducing ability of plasma, as well as decreased (p < 0.05) malondialdehyde, IL-6, IL-1ß, tumor necrosis factor (TNF-α), interferon-γ, thiobarbituric acid-reactive substances, and diamine oxidase level in serum and TNF-α level in the jejunal mucosa. Moreover, these pigs also showed enhanced (p < 0.05) villus height/crypt depth in ileum, villus height in duodenum, protein expression of zonula-occludens-1 in jejunal mucosa, and fecal total volatile fatty acids and butyric acid contents. In conclusion, ESBM replacing EFS could enhance performance via improving immune response, antioxidant status, gut morphology, and barrier function of nursery pigs in antibiotic free diets.

Changes in Growth Performance and Ileal Microbiota Composition by Xylanase Supplementation in Broilers Fed Wheat-Based Diets.

Xylanase exerts key roles in improving growth performance and intestinal health of broilers fed wheat-based diets. However, knowledge is limited regarding effects of xylanase supplementation on ileal microbiota in broilers. A total of 128 one-day-old broilers (initial BW 48.03 ± 0.33 g) were selected to investigate effects of xylanase (AT-xynA) on growth performance, ileal morphology, microbiota composition, immune response, antioxidant capacity, and endocrine peptide levels in broilers. Broilers were randomly allotted into two dietary treatments (n = 8), namely, a wheat-soybean basal diet and a basal diet with 4,000 U/kg AT-xynA (XY). On days 7, 14, 21, and 42, broilers were weighted and ileal tissues were sampled. Ileal digesta samples were collected for analyzing microbiota composition on days 21 and 42. The results showed that AT-xynA could improve average daily weight gain and average daily feed intake, and there were interactions between diet and age of broilers (p < 0.05). On days 21 and 42, xylanase supplementation decreased ileal microbiota α-diversity, and the relative abundance of potentially pathogenic microbiota, such as phylum Proteobacteria, family Moraxellaceae and Staphylococcaceae, genus Staphylococcus, Pseudomonas, Streptococcus, and Enterococcus, increased the abundance of Lactobacillus (p < 0.05). Moreover, the reduction in acetate concentration and abundance of short-chain fatty acid-producing bacteria was also observed in broilers from XY group (p < 0.05). AT-xynA increased ileal villus height, glucagon-like peptide-1, and insulin-like growth factor-1 concentrations and decreased interleukin-1ß, interleukin-6, tumor necrosis factor-α, and malondialdehyde content in broilers, and these positive effects on intestinal health were greater in young broilers. In conclusion, xylanase supplementation to wheat-based diets could improve ileal intestinal morphology and immune function, and alleviate excess fermentation of bacteria, which may be related to changes of intestinal microbiota. In addition, the positive effects of xylanase on intestinal health were more pronounced in young broilers, thus contributing to subsequent improvement in growth performance of broilers.

Isolation, structure characteristics and antioxidant activity of two water-soluble polysaccharides from Lenzites betulina.

BACKGROUND: Fungal polysaccharides belong to a very important class of biological macromolecules in nature, and have complex monosaccharide composition and structure. These studies on structure and biological activity of fungal polysaccharides have become one of the research hotspots of scholars at home and abroad. RESULTS: This study was performed in order to understand the structural characteristics and antioxidant activity of polysaccharides from Lenzites betulina (LBPs). The LBPs were deproteinized using sevag method, and further purified by DEAE cellulose-52 column and Sephadex G-100 column chromatographies, then the two refined polysaccharides were obtained and named LBPs-5 and LBPs-6. Fourier transform infrared spectrometry (FT-IR) showed that LBPs-5 and LBPs-6 are typical ß-pyranose with characteristic peaks of polysaccharides. The molecular weight of the two water-soluble polysaccharides were estimated to be 3.235 × 103 Da and 6.196 × 103 Da by HPGPC, respectively. HPLC with PMP derivatization analysis indicated that the monosaccharide compositions of LBPs-5 were mannose, glucuronic acid, glucose, and galactose in a molar ratio of 0.05:0.15:0.76:0.04. The monosaccharide compositions of LBPs-6 were mannose, glucuronic acid, and glucose, in a molar ratio of 0.04:0.17:0.79. Furthermore, the two water-soluble polysaccharides demonstrated strong scavenging effects on DPPH·, ABTS·+, ·OH and weak total reducing power, especially LBPs-6 was significantly stronger in scavenging rate than that of LBPs-5. CONCLUSIONS: The outcome of the study indicated that LBPs had good potential as medicine and food.

MiR-1254 and MEGF6 regulates oxaliplatin resistance in human colorectal cancer cells.

Colorectal cancer (CRC) remains one of the deadliest diseases in the whole world. Cancer recurrence and chemotherapeutic drug resistance limit the overall survival rate of patients with CRC. This study aimed to discover the latent miRNAs and genes associated with oxaliplatin resistance in CRC cells. The study found that miR-1254 is upregulated in oxaliplatin-resistant CRC cell line HCT116-R compared with its parental cell line HCT116 by transcriptome sequencing and small RNA sequencing. Meanwhile, MEGF6 (multiple EGF-like domains 6) was downregulated in HCT116-R cells. Transient transfection of miR-1254 mimics significantly reduced cell apoptosis, increased HCT116 tolerance to oxaliplatin, and enhanced MEGF6 expression. Furthermore, transfection of miR-1254 inhibitor increased apoptosis, decreased HCT116-R tolerance to oxaliplatin, and reduced MEGF6 expression. In addition, transient transfection of SiMEGF6 enhanced HCT116 cell resistance to oxaliplatin and reduced cell apoptosis. In summary, MEGF6 is a latent functional target of miR-1254 in regulating oxaliplatin resistance and apoptosis in human CRC cells, suggesting a potential therapeutic target for CRC.

Bioinformatics analysis combined with experiments predicts CENPK as a potential prognostic factor for lung adenocarcinoma.

BACKGROUND: Lung cancer is the most common malignant tumor. Identification of novel diagnostic and prognostic biomarkers for lung cancer is a key research imperative. The role of centromere protein K (CENPK) in cancer is an emerging research hotspot. However, the role of CENPK in the progression of lung adenocarcinoma (LAC) is not well characterized. METHODS: In this study, we identified CENPK as a potential new gene for lung cancer based on bioinformatics analysis. In addition, in vitro experiments were performed to verify the function of this gene. We investigated the expression of CENPK in LAC by analyses of datasets from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Differential expression analyses, gene ontology (GO) enrichment, Kyoto encyclopedia of genes and genomes (KEGG) analysis, and gene set enrichment analysis (GSEA) were conducted to evaluate the diagnostic and prognostic relevance of CENPK. Then, for evaluating the biological behavior and role of CENPK in lung cancer cells, we did a series of vitro experiments, such as immunohistochemistry analysis, Western blot analysis, CCK8 assay, transwell assay, flow cytometry, and wound healing assay. RESULTS: We demonstrated overexpression of CENPK in LAC; in addition, increased expression of CENPK was associated with clinical progression. Moreover, CENPK was found to be an independent risk factor in patients with LAC. Furthermore, we observed activation of CENPK-related signaling pathways in patients with LAC. CONCLUSIONS: Our findings indicate a potential role of CENPK in promoting tumor proliferation, invasion, and metastasis. It may serve as a novel diagnostic and prognostic biomarker in patients with LAC.

Delayed postoperative radiotherapy might improve the long-term prognosis of locally advanced esophageal squamous cell carcinoma.

OBJECTIVE: There is no consensus on the optimal timing of postoperative radiotherapy (PORT) for locally advanced esophageal squamous cell carcinoma (ESCC). We aimed to determine whether the timing of PORT affects the long-term prognosis of ESCC, and plotted nomograms to predict survival. METHODS: We retrospectively analyzed 351 ESCC patients who underwent radical surgery and PORT. Receiver operating characteristic curves were used to estimate the optimal cutoff point of the time interval between surgery and PORT. Cox proportional hazards regression was used to identify prognostic predictors. Overall survival (OS) and progression-free survival (PFS) were predicted using nomograms. RESULTS: The median follow-up was 53 months (range: 3-179 months). Compared to early PORT, PORT at >48 days after surgery was associated with better OS (adjusted hazard ratio [HR]: 1.406, p = 0.037) and PFS (adjusted HR: 1.475, p = 0.018). In the chemotherapy subgroup, incorporation of chemotherapy timing into the analysis suggested that 2-4 chemotherapy cycles followed by PORT was the optimal treatment schedule as compared to 0-1 chemotherapy cycle followed by PORT and concurrent chemoradiotherapy (5-year PFS: 65.9% vs. 51.0% vs. 50.1%; p = 0.049). The nomograms for OS and PFS were superior to the TNM classification (concordance indices: 0.721 vs. 0.626 and 0.716 vs. 0.610, respectively). CONCLUSIONS: Delayed PORT (>48 days) provides better survival benefit than early PORT among ESCC patients. PORT following 2-4 chemotherapy cycles might lead to the best survival rate. The nomogram plotted in this study effectively predicted survival and may help guide treatment.

LncRNA MAGI2-AS3 Overexpression Sensitizes Esophageal Cancer Cells to Irradiation Through Down-Regulation of HOXB7 via EZH2.

BACKGROUND: Accumulating evidence has suggested that aberrant expression of long non-coding RNAs (lncRNAs) may contribute to cancer progression in association with radioresistance. The current study aimed to identify the potential role of lncRNA MAGI2-AS3 and the underlying mechanism in its regulation of the radio-sensitivity of esophageal cancer cells. METHODS AND RESULTS: Initially, we detected high expression of HOXB7 from microarray-based gene expression profiling of esophageal cancer. Then, we identified the interactions among MAGI2-AS3, HOXB7, and EZH2 by dual-luciferase reporter gene assay, RNA pull-down assay, RIP assay and ChIP assay. HOXB7 was highly-expressed, while MAGI2-AS3 was poorly-expressed in esophageal cancer tissues and cells. The effect of MAGI2-AS3 and HOXB7 on esophageal cancer cell proliferation and apoptosis as well as tumorigenicity of radioresistant cells was examined by gain- and loss-of-function experiments. Interestingly, MAGI2-AS3 down-regulated HOXB7 through interaction with EZH2, which promoted cell apoptosis and inhibited proliferation and radio-resistance. Besides, down-regulation of MAGI2-AS3 exerted a promoting effect on these malignant phenotypes. CONCLUSION: Taken together, our results reveal the potential role of MAGI2-AS3 over-expression in controlling esophageal cancer resistance to radiotherapy by down-regulating HOXB7, this providing a candidate biomarker for resistance to radiotherapy.

MPEG grafted alkylated carboxymethyl chitosan as a high-efficiency demulsifier for O/W crude oil emulsions.

Three kinds of novel environmentally benign and high-efficiency crude oil demulsifiers were prepared using methoxy polyethylene glycol (MPEG) to modify alkylated carboxymethyl chitosan (ACMC). Structures of the demulsifiers were confirmed using FT-IR and 1H NMR, and the relationship between surface tension and concentration was tested. Demulsification performance was investigated using the bottle test method with oil-in-water (O/W) emulsions that were prepared in lab conditions. The demulsification efficiency was as high as 79.1 %-84.9 %, and the possible mechanism of the demulsification process is discussed. Results show that MPEG-grafted ACMC (MPEG-ACMC) has a promising application as a demulsifier for dealing with emulsified O/W crude oil.

Peptides of tetraspanin oncoprotein CD151 trigger active immunity against primary tumour and experimental lung metastasis.

BACKGROUND: Active immunotherapy is an effective, long-lasting, cheap, and safe approach to suppress cancer progression; however, the key issue is to develop appropriate tumour vaccines. Oncoproteins are up-regulated under various stress conditions and promote cell survival. Oncoproteins and their immunogenic domains could serve well as tumour vaccines and prime the hosts' active anti-tumour immunity. METHODS: Proteomic and bioinformatic analyses were performed to identify potential tumour associated antigens (TAAs). Then, peptides derived from CD151 were designed and synthesized according to the major histocompatibility complex (MHC) I binding and immunogenicity. Cytotoxicity assay, flow cytometry, immunohistochemistry, and in vivo bioluminescence imaging were performed to assess the active anti-tumour immunity triggered by CD151 peptides in H22 primary hepatoma and experimental 4T1 breast cancer lung metastasis models. FINDINGS: CD151 was identified as an ideal TAA based on proteomic and bioinformatic analyses. CD151 peptides as tumour vaccines triggered active anti-tumour immunity against H22 hepatoma and the lung metastasis of 4T1 breast cancer in two mouse models through the activation of CD8+IFNγ+ lymphocytes and the subsequent targeted cytotoxicity. Further, the peptides suppressed the negative regulators, myeloid-derived suppressor cells. Survival was prolonged for mice with lung metastases from CD151 peptide-immunised groups. INTERPRETATION: The up-regulated oncoproteins in 8 Gy-irradiated tumour cells are good candidates for designing immunogenic peptides as tumour vaccines. Anti-tumour active immunity primed by peptides from CD151 may be an effective and safe approach to suppress cancer progression.