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Sustained activation of signal transducer and activator of transcription 3 (STAT3) is a critical contributor in tumorigenesis and chemoresistance, thus making it an attractive cancer therapeutic target. Here, SH2 domain-containing adapter protein F (SHF) is identified as a tumor suppressor in glioblastoma Multiforme (GBM) and its negative regulation of STAT3 activity is characterized. Mechanically, SHF selectively binds and inhibits acetylated STAT3 dimerization without affecting STAT3 phosphorylation or acetylation. Additionally, by blocking STAT3-DNMT1 (DNA Methyltransferase 1) interaction, SHF relieves methylation of tumor suppressor genes. The SH2 domain is documented to be essential for SHF's actions on STAT3, and almost entirely replaces the functions of SHF on STAT3 independently. Moreover, the peptide C16 a peptide derived from the STAT3-binding sites of SHF inhibits STAT3 dimerization and STAT3/DNMT1 interaction, and achieves remarkable growth inhibition in GBM cells in vitro and in vivo. These findings strongly identify targeting of the SHF/STAT3 interaction as a promising strategy for developing an optimal STAT3 inhibitor and provide early evidence of the potential clinical efficacy of STAT3 inhibitors such as C16 in GBM.

We have previously demonstrated that mesenchymal stromal/stem cells (MSCs) in spheroids (MSCsubsp/sub) tolerate ambient and hypoxic conditions for a prolonged time. Local administration of MSCsubsp/sub, but not dissociated MSCs (MSCsubdiss/sub), promotes wound healing and relieves multiple sclerosis and osteoarthritis in mice and monkeys. These findings indicate an advantage of MSCsubsp/subover MSCsubdiss/subin sustaining cell viability and efficacy following transplantation, which, however, does not appear to apply to intravenous (i.v.) injection for the principal concern that MSCsubsp/submight cause embolism in small blood vessels of the host, leading to sudden death. Here, we addressed this concern by injecting human MSCsubsp/sub(∼450 μm) or MSCsubdiss/subi.v. into cynomolgus monkeys. Surprisingly, no deaths occurred until sacrifice at day 21 or 60 post injection, and no remarkable physiological changes were found in the animals following the i.v. injection. The big diameters of large blood vessels in monkeys, compared to small animals like mice, may allow sufficient time for MSCsubsp/subto dissociate into single cells so they can pass through small vessels without causing embolism. Retention of MSCsubsp/subwas lower in the lungs but higher in the blood than retention of MSCsubdiss/subat 1 h post injection and both disappeared at day 21. In vitro, MSCsubsp/subtolerated fluidic shear stress with higher survival than MSCsubdiss/sub. Thus, i.v. injection of MSCsubsp/subinto nonhuman primates is feasible, safe, and probably associated with better survival, less lung entrapment and higher efficacy than administration of MSCsubdiss/sub.

Description, Circulating tumor cells with high levels of adhesion proteins survive more easily in circulation and form metastatic tumors., To study how cancer cells can withstand fluid shear stress (SS), we isolated SS-resistant breast and lung cancer cells using a microfluidic circulatory system. These SS-resistant cells showed higher abilities to form clusters, survive in circulation, and metastasize in mice. These SS-resistant cells expressed 4.2- to 5.3-fold more desmocollin-2 (DSC2) and plakophilin-1 (PKP1) proteins. The high expression of DSC2 and PKP1 facilitated cancer cells to form clusters in circulation, and also activated PI3K/AKT/Bcl-2–mediated pathway to increase cell survival. The high levels of DSC2 and PKP1 are also important for maintaining high expression of vimentin, which stimulates fibronectin/integrin β1/FAK/Src/MEK/ERK/ZEB1–mediated metastasis. Moreover, higher levels of DSC2 and PKP1 were detected in tumor samples from patients with breast and lung cancer, and their high expression was correlated with lower overall survival and worse disease progression. DSC2 and PKP1 may serve as new biomarkers for detecting and targeting metastatic circulating tumor cells.

Cellular heterogeneity and the lack of metastatic biomarkers limit the diagnosis of and development of therapies for metastatic triple-negative breast cancer (TNBC). Thus, development of new clinically relevant markers is urgently needed. By using RNA-seq analysis, we found that nerve growth factor receptor (NGFR) was highly expressed in metastatic lung clones of MDA-MB-231 cells. This high level of NGFR expression was necessary for TNBC cells to grow into tumor spheres under nonadhesive conditions, resist anoikis, promote primary tumor growth and increase metastasis in mice. NGFR was also expressed at a high level in a greater number of TNBC patients (45%) than non-TNBC patients (23%), enriched in higher grade tumors, and negatively correlated with the overall survival of TNBC patients. Mechanistic analysis indicated that NGFR exerted its prometastatic effects by binding with neurotrophic receptor tyrosine kinase 3 (TrkC) mainly through a ligand-independent manner, which activated the MEK-ERK1-ZEB1 and PI3K-AKT signaling pathways, increased the level of fibronectin, and decreased the expression of PUMA. Notably, we observed that NGFR expression in TrkC-positive metastatic clones reduced cellular sensitivity to anti-Trk therapy. Moreover, WNT family member 5a (WNT5A) and TrkC activated NGFR transcription in a ZEB1-dependent manner. Taken together, this study identified NGFR as a novel driver for transforming TNBC into higher grade metastatic tumors. Our findings provide the basis for the future development of NGFR as a diagnostic and prognostic marker for determining the metastatic potential of TNBC and as a therapeutic target for treating TNBC patients.

This study manifested the effects of polysaccharides from Ganoderma lucidum strain S3 (GLP S3) on chronic pancreatitis (CP) therapy and intestinal microbiota modulation in mice induced by diethyldithiocarbamate (DDC). The GLPS3 was prepared from cultured mycelium and markedly alleviated the pancreatitis in mice through decreasing lipase, AMS, IFN-γ and TNF-α level as well as increasing SOD and total antioxidant activity. Furthermore, high throughput sequencing analysis revealed that GLPS3 altered the composition and diversity of intestinal microbiota, especially, decreased the relative abundance of phylum Bacteroidetes and increased that of phylum Firmictutes. At the genus level, supplementation of GLPS3 increased the relative abundance of the beneficial bacteria such as Lactobacillales, Roseburia and Lachnospiraceae. These results disclosed the potential therapy mechanism of GLPS3 on chronic pancreatitis might be intestinal microbiota dependent.

The ability of seleno-lentinan in preventing chronic pancreatitis was examined and its prebiotic-like efficacy on gut microbiota in mice induced by 3,5-diethoxycarbonyl-1,4-dihydrocollidine was also investigated. Seleno-lentinan more effectively impeded the progress of inflammation and fibrosis than selenite and lentinan through the enhancement of superoxide dismutase and glutathione peroxidase activities, inhibiting lipid peroxidation and decreasing the levels of serum tumour necrosis factor-α and interleukin-1β, as well as pancreatic hydroxyproline. The high throughput sequencing analysis showed that seleno-lentinan increased the relative abundance of Bacteroidetes, whereas it decreased that of Firmicutes. At the genus level, seleno-lentinan increased the proportion of beneficial bacteria such as Lactobacillus , Bacteroides , Prevotella , and Roseburia . Besides, the supplementation of selenium had a beneficial impact on the proliferation of lactic acid bacteria. Thus, seleno-lentinan availably prevents chronic pancreatitis development by elevating antioxidant status and has a prebiotic-like efficacy on the modulation of gut microbiota in mice.

Chronic pancreatitis (CP) is a progressive inflammation of pancreas characterized by irreversible morphologic change and dysfunction. Patients with chronic pancreatitis often present with abdominal pain, diarrhoea, jaundice, weight loss and the development of diabetes. Polysaccharides of Ganoderma lucidum strain S3 (GLPS3) possess antioxidative and immunomodulatory activities. This study was to characterize chemical structures of GLPS3 and determine their effects on diethyldithiocarbamate (DDC)-induced CP in mice. The total sugar content of GLPS3 from fermentation broth (GLPS3-Ⅰ), cultured mycelia (GLPS3-Ⅱ) and fruiting body (GLPS3-Ⅲ) was 90.4%, 92.2% and 91.8% respectively. GLPS3-Ⅰ, GLPS3-Ⅱ and GLPS3-Ⅲ were composed of Glu:Gal:Ara:Xyl, Glu:Gal:Ara:Xyl:Man:Rha, and Glu:Gal:Xyl:Man:Rha:Fuc, with molar ratio of 2.82: 1.33: 1.26: 0.87, 5.84: 2.23: 0.72:1.38: 1.40: 0.51 and 5.34: 2.72: 1.14: 1.10: 0.33: 0.38, respectively. The antioxidative activity of GLPS3-Ⅱfrom cultured mycelia in vitro is higher than other two polysaccharides. The superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in serum were increased while the malondialdehyde (MDA) levels were reversely decreased by GLPS3 treatment. Serum amylase (AMS) and lactic dehydrogenase (LDH) changes indicated the therapeutic effects of GLPS3. Moreover, interleukin-1beta (IL-1β) and interferon-gamma (INF-γ) contents were reduced most by GLPS3-Ⅱ. The results revealed that GLPS3 especially GLPS3-Ⅱfrom cultured mycelia were effective for CP therapy and bioactivity difference might be attributed to monosaccharide composition.

To remove heavy metals from aqueous solution and reclaim valuable materials from mushroom byproducts, Flammulina velutipes spent substrate (FVSS) was developed as a novel biosorbent for copper ion removal. Batch experiments demonstrated that ion removal was pH-, biosorbent dosage- and initial metal concentration dependent. The maximum removal capacity of 15.56 mg/g was achieved at pH 6.0 with a biomass dosage of 3.0 g/L and initial copper ion concentration of 50 mg/L. The adsorption data were in compliance with the Langmuir isotherm and a pseudo-second-order kinetic model. Thermodynamic studies revealed the biosorption process was endothermic, random, and spontaneous. FT-IR spectral analysis confirmed that hydroxyl, amino, carbonyl, and phosphate groups on the biosorbent surface were involved in the biosorption. The uneven surface and porous structure of the biosorbent was propitious for quickly capturing the metal ions from aqueous solution. EDX spectra revealed that the copper ions were loaded on the surface of the biosorbent. XRD patterns showed the formation of copper-containing compounds.

Inonotus obliquus polysaccharide (IOP) was extracted by water with a yield of 9.83% and purified by an anion-exchange DEAE cellulose column and Sephadex G-200 gel with a polysaccharide content of 98.6%. The scavenging activities for 2,2-diphenyl-1-picryl-hydrazyl (DPPH) and hydroxyl radicals of IOP were 82.3% and 81.3% respectively at a concentration of 5 mg/mL. IOP was composed of Man, Rha, Glu, Gal, Xyl and Ara in a molar ratio of 9.81:3.6：29.1：20.5：21.6：5.4 respectively. The gel permeation chromatography indicated that IOP was a homogeneous polysaccharide with molecular weight of 32.5 kDa. IOP helped to alleviate pancreatic acinar atrophy and weight loss for chronic pancreatitis (CP) mice induced by Diethyldithiocarbamate (DDC). The SOD level was increased most by IOP-H treatment (400 mg/kg body weight). MDA, IL-1β and LDH were significantly decreased by IOP treatment, especially hydroxyproline, IFN-γ and AMS levels were decreased 39.18%, 37.82% and 41.57% by IOP-H treatment respectively compared to MC group. In conclusion, IOP possessed strong antioxidant activity for scavenging free radicals in vitro and vivo which could be propitious to CP therapy in mice.

Lentinan was successfully modified with nitric acid-sodium selenite method based on L9(3(4)) orthogonal experiments. The optimum selenizing conditions were obtained according to selenium conversion rate as follows: Lentinan of 1.0g, pH of 4.5, temperature of 70°C and sodium selenite of 1.50g. The antioxidant activity assays in vitro (DPPH, reducing power, superoxide radicals and hydroxyl radicals) proved that Lentinan had stronger antioxidant activity after selenizing. The elevations of serum alanine aminotransferase and aspartate aminotransferase, as well as the abnormal hepatic architecture, verified that oral administration of Seleno-Lentinan (SL2-1) markedly alleviated oxidative damage in the liver of mice induced by D-gal. In addition, SL2-1 significantly increased total antioxidant capacity, activities and protein expressions of catalase and glutathione peroxidase and lowered malondialdehyde levels in serum and liver. Fourier transform infrared spectroscopy analysis indicated that selenium of SL2-1 was mostly existed as the formations of OSeO, SeO and SeOC. Scanning electron microscope coupled with energy dispersive X-ray spectroscopy analysis revealed that the surface structure and elemental components of Lentinan significantly changed after selenizing. The results are instructive for the development of organic selenium-supplement resource.