The Protective Role of Magnoliae Flos in Preventing Ovotoxicity and Managing Ovarian Function: An In Vitro and In Vivo Study.

Magnoliae Flos (MF) is a medicinal herb widely employed in traditional medicine for relieving sinusitis, allergic rhinitis, headaches, and toothaches. Here, we investigated the potential preventive effects of MF extract (MFE) against 4-vinylcyclohexene diepoxide (VCD)-induced ovotoxicity in ovarian cells and a mouse model of premature ovarian insufficiency (POI). The cytoprotective effects of MFE were assessed using CHO-K1 or COV434 cells. In vivo, B6C3F1 female mice were intraperitoneally injected with VCD for two weeks to induce POI, while MFE was orally administered for four weeks, beginning one week before VCD administration. VCD led to a significant decline in the viabilities of CHO-K1 and COV434 cells and triggered excessive reactive oxygen species (ROS) production and apoptosis specifically in CHO-K1 cells. However, pretreatment with MFE effectively prevented VCD-induced cell death and ROS generation, while also activating the Akt signaling pathway. In vivo, MFE increased relative ovary weights, follicle numbers, and serum estradiol and anti-Müllerian hormone levels versus controls under conditions of ovary failure. Collectively, our results demonstrate that MFE has a preventive effect on VCD-induced ovotoxicity through Akt activation. These results suggest that MFE may have the potential to prevent and manage conditions such as POI and diminished ovarian reserve.

Colorectal Cancer Stem Cell Subtypes Orchestrate Distinct Tumor Microenvironments.

Several classification systems have been developed to define tumor subtypes in colorectal cancer (CRC). One system proposes that tumor heterogeneity derives in part from distinct cancer stem cell populations that co-exist as admixtures of varying proportions. However, the lack of single cell resolution has prohibited a definitive identification of these types of stem cells and therefore any understanding of how each influence tumor phenotypes. Here were report the isolation and characterization of two cancer stem cell subtypes from the SW480 CRC cell line. We find these cancer stem cells are oncogenic versions of the normal Crypt Base Columnar (CBC) and Regenerative Stem Cell (RSC) populations from intestinal crypts and that their gene signatures are consistent with the "Admixture" and other CRC classification systems. Using publicly available single cell RNA sequencing (scRNAseq) data from CRC patients, we determine that RSC and CBC cancer stem cells are commonly co-present in human CRC. To characterize influences on the tumor microenvironment, we develop subtype-specific xenograft models and we define their tumor microenvironments at high resolution via scRNAseq. RSCs create differentiated, inflammatory, slow growing tumors. CBCs create proliferative, undifferentiated, invasive tumors. With this enhanced resolution, we unify current CRC patient classification schema with TME phenotypes and organization.

Berteroin ameliorates lipid accumulation through AMPK-mediated regulation of hepatic lipid metabolism and inhibition of adipocyte differentiation.

AIMS: Berteroin (5-methylthiopentyl isothiocyanate) is a naturally occurring sulforaphane analog containing a non-oxidized sulfur atom in cruciferous vegetables. The objectives of the present study were to determine the effects of berteroin on lipid metabolism in hepatocytes and adipocytes and to elucidate the mechanisms involved. MAIN METHODS: The effect of berteroin on lipid metabolism were evaluated in liver X receptor α agonist-stimulated HepG2 cells and adipocyte differentiation-induced 3T3-L1 cells using MTT assay, western blot, real time polymerase chain reaction, oil red O staining, and triglyceride assay. KEY FINDINGS: T0901317 treatment increased the expression of sterol regulatory element binding protein (SREBP)-1c, a major transcription factor that mediates lipogenesis, and berteroin pretreatment significantly inhibited the expressions of T0901317-induced SREBP-1c and lipogenic genes. Especially, berteroin had a greater inhibitory effect on T0901317-induced SREBP-1c activation than sulforaphane, AICAR, or metformin. Berteroin also markedly suppressed lipid droplet formations and triglyceride accumulations caused by both T0901317 stimulation in HepG2 hepatocytes and differentiation induction in 3T3-L1 preadipocytes. However, berteroin significantly increased the expression of mitochondrial fatty acid oxidation-related genes (carnitine palmitoyltransferase 1 (CPT-1) and peroxisome proliferator-activated receptor gamma coactivator-1α) and the phosphorylation of adenosine monophosphate-activated protein kinase (AMPK) in HepG2 cells. Interestingly, effects of berteroin on the expressions of SREBP-1c protein and CPT-1 mRNA were remarkably prevented by compound C (an AMPK inhibitor). SIGNIFICANCE: Our results suggest berteroin-inhibited hepatic lipid accumulation and adipocyte differentiation might be mediated by AMPK activation and that berteroin might be useful for the prevention, amelioration, and treatment of metabolic diseases, including hepatic steatosis.

Anti-Neuroinflammatory Effects and Mechanism of Action of Fructus ligustri lucidi Extract in BV2 Microglia.

For centuries, Fructus ligustri lucidi (FLL; the fruit of Ligustrum lucidum Aiton or Ligustrum japonicum Thunb.) has been commonly used in traditional Chinese medicine for treating hepatitis and aging-related symptoms and in traditional Korean medicine to detoxify kidneys and the liver. Pharmacological research has shown FLL has antioxidant, anti-inflammatory, anticancer, anti-osteoporosis, and hepatoprotective activities. This study was undertaken to investigate the effects of FLL extract (FLLE) on neuroinflammation. After setting a non-toxic concentration using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] assay data, we investigated the effects of FLLE using Western blotting, cell migration, enzyme-linked immunosorbent assay, a nitric oxide (NO) assay, and immunofluorescence staining in lipopolysaccharide (LPS)-stimulated murine BV2 microglial cells. FLLE was non-toxic to BV2 cells up to a concentration of 500 µg/mL and concentration-dependently inhibited the production of NO and prostaglandin E2 and the protein levels of inducible nitric oxide synthase and cyclooxygenase-2 under LPS-induced inflammatory conditions. It also inhibited the secretion of the inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Furthermore, FLLE pretreatment attenuated LPS-induced increases of CD68 (a marker of microglia activation) and suppressed the activation of mitogen-activated protein kinases (MAPKs) and nuclear factor-kappa B (NF-κB) signaling pathways in LPS-stimulated BV2 cells, and significantly increased heme oxygenase (HO)-1 levels. FLLE also reduced the LPS-induced increase in the migratory ability of BV2 cells and the phosphorylation of vascular endothelial growth factor receptor 1. Collectively, FLLE effectively inhibited inflammatory response by suppressing the MAPK and NF-κB signaling pathways and inducing HO-1 in LPS-stimulated BV2 microglial cells. Our findings provide a scientific basis for further study of FLL as a candidate for preventing or alleviating neuroinflammation.

HS­146, a novel phosphoinositide 3­kinase α inhibitor, induces the apoptosis and inhibits the metastatic ability of human breast cancer cells.

The phosphoinositide 3­kinase (PI3K) signaling pathway plays an important role in human cancer as it regulates critical cellular functions, such as survival, proliferation and metabolism. In the present study, a novel PI3Kα inhibitor (HS­146) was synthesized and its anticancer effects on MCF­7, MDA­MB­231, SKBR3 and BT­474 human breast cancer cell lines were confirmed. HS­146 was found to be most effective in inhibiting the proliferation of MCF­7 cells and in inducing cell cycle arrest in the G0/G1 phase by downregulating cyclin D1, cyclin E, cyclin­dependent kinase (Cdk)2 and Cdk4, and upregulating p21Waf1/Cip1 protein levels in this cell line. The induction of apoptosis by HS­146 was confirmed by DAPI staining and western blot analysis. Cell shrinkage and nuclear condensation, which are typical morphological markers of apoptosis, were increased by HS­146 in the MCF­7 cells in a concentration­dependent manner, and HS­146 also increased the protein expression levels of cleaved poly(ADP­ribose) polymerase (PARP) and decreased the protein expression levels of Mcl­1 and caspase­7. In addition, HS­146 effectively decreased the phosphorylation levels of downstream PI3K effectors, such as Akt, mammalian target of rapamycin (mTOR), glycogen synthase kinase 3ß (GSK3ß), p70S6K1 and eukaryotic translation initiation factor 4E­binding protein 1 (4E­BP1). Hypoxia­inducible factor (HIF)­1α and vascular endothelial growth factor (VEGF) expression were also suppressed by HS­146 under hypoxic conditions, and HS­146 inhibited the migration and invasion of MCF­7 cells in a concentration­dependent manner. On the whole, the findings of the present study suggest that HS­146, a novel PI3Kα inhibitor, may be an effective novel therapeutic candidate that suppresses breast cancer proliferation and metastasis by inhibiting the PI3K/Akt/mTOR pathway.

Anticancer Effects of a Korean Herbal Medicine Formula (H9) via AMPK and HER2-PI3K/Akt Signaling in Breast Cancer Cells.

An H9 is a formula of nine medicinal herbs derived from Osuyubujaijung-tang, a traditional Korean prescription for Soeumin constitution. In our previous study, H9 showed anticancer effects against breast cancer and non-small-cell lung cancer. However, the underlying mechanisms of these effects have not yet been elucidated. In this study, we investigated the effects of H9, both alone and in combination with trastuzumab, on breast cancer cells and sought to elucidate the mechanisms involved. H9 suppressed the proliferation of human breast cancer cells, induced arrest of the cell cycle at the G0/G1 phase, and caused mitochondrial dysfunction and apoptosis. In addition, H9 induced the activation of AMPK and inhibited the HER2-PI3K/Akt signaling pathway. Furthermore, H9 attenuated hypoxia-induced HIF-1α and VEGF, resulting in decreased migration and invasion of breast cancer cells. Compared with treatment with either drug alone, co-treatment with H9 and trastuzumab significantly inhibited the growth of BT-474 cells through induction of apoptosis. These results suggest that H9 should be considered as a potent anticancer agent that targets the HER2-PI3K/Akt pathway, and the combination of H9 with trastuzumab should be considered as a new therapeutic regimen for treating breast cancer. Copyright © 2017 John Wiley & Sons, Ltd.

Speckle noise reduction in ultrasound images using a discrete wavelet transform-based image fusion technique.

Here, the speckle noise in ultrasonic images is removed using an image fusion-based denoising method. To optimize the denoising performance, each discrete wavelet transform (DWT) and filtering technique was analyzed and compared. In addition, the performances were compared in order to derive the optimal input conditions. To evaluate the speckle noise removal performance, an image fusion algorithm was applied to the ultrasound images, and comparatively analyzed with the original image without the algorithm. As a result, applying DWT and filtering techniques caused information loss and noise characteristics, and did not represent the most significant noise reduction performance. Conversely, an image fusion method applying SRAD-original conditions preserved the key information in the original image, and the speckle noise was removed. Based on such characteristics, the input conditions of SRAD-original had the best denoising performance with the ultrasound images. From this study, the best denoising technique proposed based on the results was confirmed to have a high potential for clinical application.

Comparison of epidermal/dermal damage between the long-pulsed 1064 nm Nd:YAG and 755 nm alexandrite lasers under relatively high fluence conditions: quantitative and histological assessments.

OBJECTIVE: The purpose of this study was to compare degrees of epidermal/dermal tissue damage quantitatively and histologically after laser irradiation, to find ideal treatment conditions with relatively high fluence for skin rejuvenation. BACKGROUND DATA: A number of recent studies have evaluated the clinical efficacy and safety of therapeutic lasers under relatively low fluence conditions. METHODS: We transmitted the long-pulsed 1064 nm Nd:YAG and 755 nm Alexandrite lasers into pig skin according to different fluences and spot diameters, and estimated epidermal/dermal temperatures. Pig skin specimens were stained with hematoxylin and eosin for histological assessments. The fluence conditions comprised 26, 30, and 36 J/cm2, and the spot diameter conditions were 5, 8, and 10 mm. Pulse duration was 30 ms for all experiments. RESULTS: Both lasers produced reliable thermal damage on the dermis without any serious epidermal injuries, under relatively high fluence conditions. The 1064 nm laser provided more active fibrous formations than the 755 nm laser, while higher risks for tissue damages simultaneously occurred. CONCLUSIONS: The ideal treatment conditions for skin rejuvenation were 8 mm diameter with 30 J/cm2 and 10 mm diameter with 26 J/cm2 for the 1064 nm laser, and 8 mm diameter with 36 J/cm2 and 10 mm diameter with 26 J/cm2 for the 755 nm laser.