Distinctions in structure, rheology, antioxidation, and α-glucosidase inhibitory activity of ß-glucans from different species.

To investigate the distinctions between ß-glucans from different species, Lentinula edodes ß-glucan (LG), yeast ß-glucan (YG), and oat ß-glucan (OG) were extracted with hot water and determined as ß-d-glucopyranose form by HPLC and FT-IR analysis. The molecular weight (Mw) of LG, YG, and OG was 670 kDa, 341 kDa, and 66 kDa, respectively. Scanning electron microscopy exhibited different micro surfaces of three ß-glucans and the relative crystallinity of YG was the highest (29.8 %), followed by that of LG (23.2 %) and OG (20.3 %) determined by X-ray diffraction. Congo red analysis and atomic force microscopy showed that LG and YG have triple helical structures. The apparent viscosity, storage modulus (G'), and loss modulus (G") of ß-glucans were increased with the increase of Mw. DPPH·, ABTS+·, HO·, and reducing power assays showed that ß-glucans from different species exhibited different antioxidant activities, and the DPPH· scavenging rate of 2 mg/mL LG reached >80 % higher than that of YG and OG. The α-glucosidase inhibitory activity of OG was better than YG and LG. In summary, ß-glucans from different species have different structures, physicochemical properties, and physiological functions, which provides theoretical evidence for the precise processing and utilization of ß-glucan.

Single-cell transcriptome analysis indicates fatty acid metabolism-mediated metastasis and immunosuppression in male breast cancer.

Male breast cancer (MBC) is a rare but aggressive malignancy with cellular and immunological characteristics that remain unclear. Here, we perform transcriptomic analysis for 111,038 single cells from tumor tissues of six MBC and thirteen female breast cancer (FBC) patients. We find that that MBC has significantly lower infiltration of T cells relative to FBC. Metastasis-related programs are more active in cancer cells from MBC. The activated fatty acid metabolism involved with FASN is related to cancer cell metastasis and low immune infiltration of MBC. T cells in MBC show activation of p38 MAPK and lipid oxidation pathways, indicating a dysfunctional state. In contrast, T cells in FBC exhibit higher expression of cytotoxic markers and immune activation pathways mediated by immune-modulatory cytokines. Moreover, we identify the inhibitory interactions between cancer cells and T cells in MBC. Our study provides important information for understanding the tumor immunology and metabolism of MBC.

Rab1A-Mediated Exosomal Sorting of miR-200c Enhances Breast Cancer Lung Metastasis.

Background: Recent therapeutic approaches have improved survival rate for women with breast cancer, but the survival rate for metastatic breast cancer is still low. Exosomes released by various cells are involved in all steps of breast cancer development. Methods: We established the multimodal imaging report expression in breast cancer cells with lentivirus vectors pGluc and pBirA to investigate the secreted exosomes. Comparative microRNA (miRNA) analysis was performed with miRNA qPCR array in mice with breast cancer lung metastasis. The co-immunoprecipitation and chromatin immunoprecipitation assays were used to identify the mechanism of miRNA sorting to exosomes. The potential therapeutic strategy using an anti-sorting antibody was used to investigate breast cancer lung metastasis. Results: We identified 26 high- and 32 low-expression level miRNAs in exosomes from metastasis compared to those from primary tumors and normal tissues. The tumor suppressors, including miR-200c and let-7a, were reduced in tumor tissues and metastasis but increased in the respective exosomes compared to normal tissues. Furthermore, the Ras-related protein (Rab1A) facilitated miR-200c sorting to exosomes circumventing the influence of tumor suppressor miR-200c on tumor cells, while the metastatic exosome cargo miR-200c inhibited F4/80+ macrophage immune response. Administration of anti-Rab1A antibody significantly repressed the trafficking of miR-200c to exosomes and breast cancer lung metastasis. Conclusion: Our study has identified a novel molecular mechanism for breast cancer lung metastasis mediated by exosome cargo miRNAs and provided a new therapeutic strategy for cancer immunotherapy.

Lactobacillus rhamnosus GG alleviates colitis caused by chemotherapy via biofilm formation.

BACKGROUND AND AIM: Severe colitis is a common side effect of chemotherapy in cancer patients. In this study, we attempted to enhance the viability of probiotics in a gastric acid environment and improve the colitis induced by dextran sulfate sodium (DSS) and docetaxel. METHODS: We purified Lactobacillus from yogurt and estimated their growth at pH 6.8 and pH 2.0. In the further investigation, the bacterial biofilm formation was used to define the mechanism by which administration of Lactobacillus rhamnosus (LGG) via oral gavage alleviates the colitis and intestine permeability of the mice induced by DSS and docetaxel. The potential benefit of probiotics on the treatment of breast cancer metastasis has been assessed as well. RESULTS: Lactobacillus from yogurt growth was unexpectedly faster in the pH 2.0 than in the neutral pH medium during the first hour. LGG administered in the fasting state via oral gavage significantly improved the preventive effect in the colitis caused by DSS and docetaxel. LGG reduced the permeability of the intestine and decreased the expression of proinflammatory cytokines, TNF-α, IL-1ß, and IL-6, in colitis by biofilm formation. Increasing the docetaxel dose may reduce breast tumor growth and metastasis in the lung but did not benefit survival due to severe colitis. However, the LGG supplement significantly improved the survival of tumor-bearing mice following a high dose of docetaxel treatment. CONCLUSIONS: Our findings provide new insights into the potential mechanism of probiotic protection of the intestine and provide a novel therapeutic strategy to augment the chemotherapeutic treatment of tumors.

Preparation and properties of water-in-oil shiitake mushroom polysaccharide nanoemulsion.

Shiitake mushroom (Lentinus edodes) polysaccharide (SMP), an important immunomodulatory substance, is usually hard to be absorbed by the intestinal tract, because of its macromolecule characteristics. The objective of this study was to develop the water-in-oil (W/O) nanoemulsion to improve its absorption efficiency and the biological activity. Based on the results of pseudoternary phase diagram, the formulation of nanoemulsion were optimized as IPM/polysaccharide/Tween80-Span85/anhydrous ethanol = 39/16/33.75/11.25 (v/v/v/v), used as the oil, aqueous phase, surfactant and co-surfactant, respectively. Under this condition, the droplet size of a W/O SMP nanoemulsion was 144.5 nm, with the polydispersity of 0.128 and the great stability. Moreover, the anti-tumor activity of SMP was notably improved after nano-emulsification. The results demonstrated that SMP nanoemulsion has great potential for applying in health food and pharmaceutical industry.

Tumor-derived nanovesicles promote lung distribution of the therapeutic nanovector through repression of Kupffer cell-mediated phagocytosis.

Tumor-derived nanovesicles have been widely used as a biomarker or therapeutic target in various tumor types. However, these nanovesicles have limited use in therapy due to the risk of advancing tumor development. Methods: Exosome-like nanovesicles (ENVs) were developed from metastatic breast cancer 4T1 cells-derived exosomes. The distribution of ENVs and their impact on macrophage-mediated phagocytosis were evaluated. The effect of ENVs pretreatment on anti-lung metastasis therapeutic effects of chemotherapeutic drugs delivered by DOTAP/DOPE liposomes in breast cancer-bearing mice was also examined. Results: We demonstrated that, following intravenous injection in mice, ENVs were preferentially uptaken by Kupffer cells and repressed phagocytosis. The decreased uptake appeared to be due to the translocation of membrane nucleolin from the inner face of the plasma membrane to the cell surface and intercellular Ca2+ fluxes, leading to altered expression of genes involved in phagocytosis by macrophages. Mice pretreated with 4T1-derived ENVs led to the decreased uptake of DOTAP: DOPE liposomes (DDL) in the liver. Consequently, doxorubicin-loaded DDL transported to the lungs instead of the liver, effectively inhibiting breast cancer lung metastasis. Importantly, 4T1 cells exosome-derived ENVs had no detectable toxicity in vivo and low-risk to promote tumor growth and metastasis compared to 4T1 cells exosomes. Conclusion: Our results suggested that pretreatment with 4T1 ENVs represents a strategy to escape Kupffer cell-mediated phagocytosis effectively targeting drug delivery vehicles to tumor metastasis, reducing the IC50 of the chemotherapeutic drugs, and avoiding adverse side effects.

Paeoniflorin influences breast cancer cell proliferation and invasion via inhibition of the Notch­1 signaling pathway.

Breast cancer is one of the most frequently occurring malignant tumors affecting women's health. At least one million new cases are diagnosed each year. Therefore, research that aims to identify strategies that inhibit the growth of breast cancer cells has become a primary worldwide focus. Traditional Chinese medicine (TCM) is regarded as a valuable resource in China, and numerous monomer compositions extracted from TCMs have been demonstrated to exhibit antitumor effects. The present study aimed to determine the impact of paeoniflorin (PF) on breast cancer cell proliferation and invasion, and to explore the mechanisms underlying its effects. Different concentrations of PF were applied to MCF­7 cells at various time points and the Cell Counting kit­8 assay was used to determine cell proliferation, a transwell invasion assay was employed to determine cell invasion, reverse transcription­polymerase chain reaction was used to determine notch homolog­1 (NOTCH­1) and Hes family basic helix­loop helix transcription factor (HES)­1 mRNA expression levels, and western blotting was used to determine NOTCH­1 and HES­1 protein expression levels. The results demonstrated that PF inhibited the proliferation of MCF­7 cells in a dose­ and time­dependent manner. Following treatment with different concentrations of PF, the total number of cells present in the PF­treated groups was significantly lower when compared with the untreated control group (P<0.05). With increasing doses of PF, the rate of cell invasion significantly decreased, indicating a dose­dependent association. NOTCH­1 and HES­1 mRNA expression levels were reduced when compared with the untreated control group, which reached a statistical significance following treatment with 15 and 30 µM PF (P<0.05). NOTCH­1 and HES­1 protein levels demonstrated a similar trend to the mRNA levels, whereby an increase in the concentration of PF was associated with a decrease in NOTCH­1 and HES­1 protein expression levels. The results of the present study therefore suggest that PF may inhibit the proliferation and invasiveness of breast cancer cells via inhibition of the NOTCH­1 signaling pathway.

MiR-30e inhibits tumor growth and chemoresistance via targeting IRS1 in Breast Cancer.

MicroRNA-30e (miR-30e) is downregulated in various tumor types. However, its mechanism in inhibiting tumor growth of breast cancer remains to be elucidated. In this study, we found that miR-30e was significantly downregulated in tumor tissues of breast cancer (BC) patients and cell lines, and overexpression of miR-30e inhibited cell proliferation, migration and invasion. To understand the potential mechanism of miR-30e in inhibiting tumor growth, we showed that miR-30e blocked the activation of AKT and ERK1/2 pathways, and the expression of HIF-1α and VEGF via directly targeting IRS1. Moreover, miR-30e regulates cell proliferation, migration, invasion and increases chemosensitivity of MDA-MB-231 cells to paclitaxel by inhibiting its target IRS1. MiR-30e also inhibited tumor growth and suppressed expression of IRS1, AKT, ERK1/2 and HIF-1α in mouse xenograft tumors. To test the clinical relevance of these results, we used 40 pairs of BC tissues and adjacent normal tissues, analyzed the levels of miR-30e and IRS1 expression in these tissues, and found that miR-30e levels were significantly inversely correlated with IRS1 levels in these BC tissues, suggesting the important implication of our findings in translational application for BC diagnostics and treatment in the future.

Overexpression of L1 cell adhesion molecule correlates with aggressive tumor progression of patients with breast cancer and promotes motility of breast cancer cells.

BACKGROUND AND PURPOSE: L1 cell adhesion molecule (L1CAM) has been observed to be aberrantly expressed and implicated in progression of several types of human cancers. However, its roles in breast cancer have not been fully elucidated. In this study, we aimed to investigate the clinical significance of L1CAM in human breast cancer and to validate whether it participates in cancer cell migration and invasion. METHODS: Immunohistochemical analysis of 100 breast cancer and matched non-cancerous breast tissues was performed to detect the expression and sub-cellular localization of L1CAM protein. Its associations with clinicopathological characteristics of breast cancer patients were statistically analyzed and its phenotypic effects were also evaluated in vitro. RESULTS: Of the 100 breast cancer patients, 89 (89.0%) were positive for L1CAM immunostaining localized in the membrane of cancer cells. The immunoreactive scores of L1CAM protein in breast cancer tissues were significantly higher than those in matched non-cancerous breast tissues (P<0.05). Chi-Square analysis showed the significant associations between L1CAM overexpression and high tumor stage (P=0.01), advanced tumor grade (P=0.03), positive lymph node metastasis (P=0.01) and tumor recurrence (P=0.01) in breast cancer patients. Moreover, we found that RNA interference-mediated knockdown of L1CAM could inhibit the migration and invasion abilities of breast cancer cells in vitro. CONCLUSIONS: Our results suggest that the overexpression of L1CAM may be related to several established markers of poor prognosis in breast cancer patients. L1CAM might be a potential therapeutic target against metastatic breast cancer.

Targeted delivery of doxorubicin to breast cancer cells by aptamer functionalized DOTAP/DOPE liposomes.

Doxorubicin is used to treat numerous types of tumors including breast cancer, yet dose-associated toxicities limit its clinical application. Here, we demonstrated a novel strategy by which to deliver doxorubicin to breast cancer cells by conjugating cancer cell-specific single-strand DNA aptamers with doxorubicin-encapsulated DOTAP:DOPE nanoparticles (NPs). We utilizing a whole-cell-SELEX strategy, and 4T1 cells with high invasive and metastatic potential were used as target cells, while non-invasive and non-metastatic 67NR cells were used as subtractive cells. Ten potential aptamers were generated after multi-pool selection. Studies on the selected aptamers revealed that SRZ1 had the highest and specific binding affinity to 4T1 cells. Then we developed SRZ1 aptamer-carried DOTAP:DOPE-DOX NPs. In vitro uptake results which were conducted by FACS indicated that the aptamer significantly promoted the uptake efficiency of DOTAP:DOPE-DOX NPs by 4T1 cells. ATPlite assay was performed to test 4T1, 67NR and NMuMG cell viability after treatment with free doxorubicin, DOTAP:DOPE-DOX particles and aptamer­loaded DOTAP:DOPE-DOX particles. As expected, the aptamers effectively enhanced accumulation of doxorubicin in the 4T1 tumor tissues as determined by in vivo mouse body images and biodistribution analysis. Consistent with the in vitro findings, aptamer-conjugated doxorubicin-loaded DOTAP:DOPE particles markedly suppressed tumor growth and significantly increased the survival rate of 4T1 tumor-bearing mice. These studies demonstrated that aptamer SRZ1 could be a promising molecule for chemotherapeutic drug targeting deliver.

A cross sectional study between the prevalence of chronic pain and academic pressure in adolescents in China (Shanghai).

BACKGROUND: The purpose of this study was to investigate the prevalence of four types of chronic pain (headache, abdominal pain, neck and shoulder pain (NSP), and low back pain (LBP)) and to explore the relationship between the prevalence of chronic pain and self-reported academic pressure in high school students in Shanghai, China. METHOD: Three thousand students were randomly surveyed on related issues using a questionnaire, and the results were analyzed using a multivariate logistic regression model. RESULTS: Among the 2849 high school students who completed the questionnaire, the overall prevalence rates of headache, abdominal pain, NSP, and LBP were 30.3, 20.9, 32.8, and 41.1%, respectively. The students in general experienced a heavy burden of learning, a high level of stress, and sleep deprivation, which were closely related to the four types of chronic pain. CONCLUSION: Chronic pain is a common condition in Chinese adolescents and is closely related to self-reported academic pressure.

Investigation of a new flux-modulated permanent magnet brushless motor for EVs.

This paper presents a flux-modulated direct drive (FMDD) motor. The key is to integrate the magnetic gear with the PM motor while removing the gear inner-rotor. Hence, the proposed FMDD motor can achieve the low-speed high-torque output and high-speed compact design requirements as well as high-torque density with a simple structure. The output power equation is analytically derived. By using finite element analysis (FEA), the static characteristics of the proposed motor are obtained. Based on these characteristics, the system mathematical model can be established. Hence, the evaluation of system performances is conducted by computer simulation using the Matlab/Simulink. A prototype is designed and built for experimentation. Experimental results are given to verify the theoretical analysis and simulation.

microRNA-200c downregulates XIAP expression to suppress proliferation and promote apoptosis of triple-negative breast cancer cells.

Despite advances in the understanding of breast cancer, patients most commonly have a poor prognosis, particularly those with triple negative breast cancer (TNBC). microRNAs (miRNAs) are endogenous non­coding small RNAs, and their aberrant expression is linked to numerous malignancies. In the present study, the expression levels of miR­200c in patients with TNBC were analyzed and it was identified that miR­200c was downregulated in TNBC samples, compared with that in normal adjacent tissues. miR­200c was overexpressed in the TNBC cell line MDA­MB­231 and its functions were studied in vitro and in vivo. An in vitro study revealed that the overexpression of miR­200c inhibited MDA­MB­231 cell proliferation and resulted in the induction of apoptosis. The in vivo data indicated that the overexpression of miR­200c significantly inhibited tumor growth and increased the rate of apoptosis. Target prediction revealed that the X­linked inhibitor of apoptosis (XIAP) had putative complementary sequences to miR­200c, which was confirmed by a dual luciferase reporter assay. Western blot analysis further demonstrated that the expression of XIAP was markedly reduced and that caspase­3 was highly activated by the overexpression of miR­200c. These findings suggested that miR­200c may function as a tumor suppressor gene in TNBC, at least partly via directly targeting XIAP, and may therefore act as a potential therapeutic target in the development of novel treatment strategies for TNBC.