GDF11 knockdown downregulates SMURF1 to inhibit breast cancer progression by activation of p53 and inactivation of ERα signaling.

Breast cancer (BC) is a prevalent neoplasm that occurs in women all over the world. Growth and differentiation factor 11 (GDF11) plays an essential role in cancer progression. This study focused on investigating the biological role and underlying mechanisms of GDF11 in BC. We detected the expression of GDF11 in 27 patients with BC and BC cell lines. Kaplan-Meier plotter was employed to analyze the relationship between GDF11 expression and overall survival (OS) of BC patients. The proliferative, migratory, invasive, and apoptotic abilities of T47D cells were examined. Correlation analysis of GDF11 with Smad ubiquitination regulatory factor 1 (SMURF1) was conducted. The association between GDF11 and the p53 pathway was analyzed by western blot and PFT-α (a p53 inhibitor)-mediated rescue assays. A brief analysis of the role of estrogen receptor alpha (ERα) signaling in BC progression was performed. The results showed that GDF11 was increased in BC tissues and cell lines, and the high expression of GDF11 was associated with the poor OS of BC patients. GDF11 knockdown inhibited the proliferation, migration, and invasion of T47D cells, but promoted cell apoptosis. Meanwhile, the GDF11 knockdown reduced the SMURF1 expression and invoked the p53 pathway activation. SMURF1 overexpression and PFT-α partially blocked the effects of GDF11 knockdown. In addition, GDF11 knockdown and SMURF1 silencing inhibited the activation of the ERα signaling pathway. In summary, GDF11 was involved in the progression of BC by regulating SMURF1-mediated p53 and ERα pathways, opening up a new way for BC treatment.

Neuroendocrine-Immune Regulatory Network of Eucommia ulmoides Oliver.

Eucommia ulmoides Oliver (E. ulmoides) is a popular medicinal herb and health supplement in China, Japan, and Korea, and has a variety of pharmaceutical properties. The neuroendocrine-immune (NEI) network is crucial in maintaining homeostasis and physical or psychological functions at a holistic level, consistent with the regulatory theory of natural medicine. This review aims to systematically summarize the chemical compositions, biological roles, and pharmacological properties of E. ulmoides to build a bridge between it and NEI-associated diseases and to provide a perspective for the development of its new clinical applications. After a review of the literature, we found that E. ulmoides has effects on NEI-related diseases including cancer, neurodegenerative disease, hyperlipidemia, osteoporosis, insomnia, hypertension, diabetes mellitus, and obesity. However, clinical studies on E. ulmoides were scarce. In addition, E. ulmoides derivatives are diverse in China, and they are mainly used to enhance immunity, improve hepatic damage, strengthen bones, and lower blood pressure. Through network pharmacological analysis, we uncovered the possibility that E. ulmoides is involved in functional interactions with cancer development, insulin resistance, NAFLD, and various inflammatory pathways associated with NEI diseases. Overall, this review suggests that E. ulmoides has a wide range of applications for NEI-related diseases and provides a direction for its future research and development.

Regulation of the NLRP3 inflammasome with natural products against chemical-induced liver injury.

The past decades have witnessed significant progress in understanding the process of sterile inflammation, which is dependent on a cytosolic complex termed the nucleotide-binding oligomerization domain (NOD)-like receptor containing pyrin domain 3 (NLRP3) inflammasome. Activation of NLRP3 inflammasome requires two steps, including the activation of Toll-like receptor (TLR) by its ligands, resulting in transcriptional procytokine and inflammasome component activation, and the assembly and activation of NLRP3 inflammasome triggered by various danger signals, leading to caspase-1 activation, which could subsequently cleave procytokines into their active forms. Metabolic disorders, ischemia and reperfusion, viral infection and chemical insults are common pathogenic factors of liver-related diseases that usually cause tissue damage and cell death, providing numerous danger signals for the activation of NLRP3 inflammasome. Currently, natural products have attracted much attention as potential agents for the prevention and treatment of liver diseases due to their multitargets and nontoxic natures. A great number of natural products have been shown to exhibit beneficial effects on liver injury induced by various chemicals through regulating NLRP3 inflammasome pathways. In this review, the roles of the NLRP3 inflammasome in chemical-induced liver injury (CILI) and natural products that exhibit beneficial effects in CILI through the regulation of inflammasomes were systematically summarized.

The synthesis of justicidinoside B and its atropisomer.

The first synthesis of justicidinoside B and its atropisomer was reported and their absolute configurations were determined by the CD exciton chirality method. The structures were confirmed by (1)H NMR, (13)C NMR, and HRMS.

Furanodiene enhances tamoxifen-induced growth inhibitory activity of ERa-positive breast cancer cells in a PPARγ independent manner.

Herbal plants are enriched with compounds with a wide range of biological activities. Furanodiene is a sesquiterpene isolated from Rhizoma Curcumae. Growing evidence shows furanodiene exhibits diversified activities of hepatoprotection, anti-inflammation, anti-angiogenesis, and anti-tumor. However, its biological activities against breast cancer have not been deeply understood, and its potential as an anti-breast cancer agent combined with tamoxifen (TAM) has not been evaluated so far. This study describes the combined effects of furanodiene and TAM in human breast cancer cells in vitro. The results showed that ERa-negative MDA-MB-231 cells were much more sensitive than ERa-positive MCF-7 cells to the growth inhibition due to furanodiene. Combined administration of furanodiene and TAM led to marked increase in growth inhibition, cell cycle arrest and pro-apoptotic activity in ERa-positive cells compared to individual agent, and enhanced the down-regulation of p-cyclin D1, cyclin D1, CDK2, CDK6, p-Rb, Rb and p-p44, and the up-regulation of p27, Bax and Bad, but did not show increased cytotoxicity in ERa-negative MCF-10A non-tumorigenic breast epithelial cells. Co-incubation induced the typical PARP cleavage or caspase 9 cleavages compared to individual agent. In addition, PPARγ activity inhibition by its antagonist T0070907 did not significantly reverse the enhanced effect of furanodiene and TAM suggesting that anti-cancer properties of combination were PPARγ independent. Our data indicated that furanodiene could enhance the growth inhibitory and pro-apoptotic activity of TAM by inducing cell cycle arrest and cell apoptosis via CDKs-cyclins and mitochondria-caspases-dependent, and PPARγ-independent signaling pathways in breast cancer cells, without contributions to the cytotoxicity of TAM.

Identification of ecdysone response elements (EcREs) in the Bombyx mori cathepsin D promoter.

Bombyx mori Cathepsin D (BmCatD) is specifically expressed in the fat body, and plays a critical role for the programmed cell death of the larval fat body and pupal gut during metamorphosis. To better understand the transcriptional control of BmCatD expression, we conducted this study to identify the ecdysone response elements (EcREs) in the BmCatD promoter and clarify their regulational functions. We inserted EcREs into a recombinant AcMNPV (Autographa californica multiple nucleopolyhedrovirus) vector and performed luciferase assay with a dual-luciferase quantitative assay system. Three putative EcREs were located at positions -109 to -99, -836 to -826 and -856 to -846 relative to the transcription start site. Overlapping deletion studies of this EcRE region showed that the three EcREs could suppress the ectopic expression of the BmCatD promoter. EcRE mutations resulted in the loss of the fat body-specific expression of the BmCatD gene. These results suggest that the EcREs are vital for activation of the promoter by 20-hydroxyecdysone (20E) in the larval fat body and further support the crucial role of ecdysone signaling to control cathepsin D gene transcription. It may suggest that the heterodimeric complex EcR/USP mediates the activation of ecdysone-dependent BmCatD transcription in the larval fat body of B. mori.

In vitro fecal fermentation properties of polysaccharides from Tremella fuciformis and related modulation effects on gut microbiota.

Tremella fuciformis is an edible and medicinal mushroom. Polysaccharides from T. fuciformis have received increasing attention due to their diversely pharmacological activities. In this study, the digestive behavior and fermentation characteristics of T. fuciformis polysaccharides (TFP) were studied. The results revealed that the reducing sugar content, chemical composition, molecular weight, rheological property, constituent monosaccharide, and FT-IR spectrum of TFP were not altered after the in vitro simulated digestion, indicating that it was indigestible under different simulated digestion conditions. However, the physicochemical characteristics of TFP, including reducing sugar content, molecular weight, constituent monosaccharide, and free monosaccharide released, were obviously altered after the in vitro fermentation for 48 h, indicating that it was remarkably utilized by intestinal microbiota in human feces. Notably, TFP could obviously modulate the microbial composition via promoting the relative abundances of Phascolarctobacterium, Bacteroides, and Lachnoclostridium. Moreover, TFP could also increase the production of short-chain fatty acids, including acetic, propionic, n-butyric, and n-valeric acids, after the in vitro fermentation for 48 h. These results showed that TFP was stable under the simulated digestion conditions, but could be utilized by intestinal microbiota in human feces, and might possess the potential to improve intestinal health.

In vitro digestive characteristics and microbial degradation of polysaccharides from lotus leaves and related effects on the modulation of intestinal microbiota.

Polysaccharides exist as one of the most abundant components in lotus leaves, which attract increasing attention owing to their promising health-promoting benefits. In this study, the digestive and microbial degradation characteristics of lotus leaf polysaccharides (LLP) were studied by using an in vitro gastrointestinal model. The results suggested that LLP was stable in the human upper gastrointestinal tract in vitro according to its digestive stabilities at different simulated digestion stages. Conversely, the indigestible LLP (LLPI) could be remarkably utilized by intestinal microbiota in human feces during in vitro fermentation, and its fermentability was 58.11% after the in vitro fermentation of 48 h. Indeed, the microbial degradation characteristics of LLPI during in vitro fermentation by human fecal inoculum were revealed. The results showed that the content of reducing sugars released from LLPI obviously increased from 0.498 to 2.176 mg/mL at the initial fermentation stage (0-6 h), and its molecular weight sharply decreased from 4.08 × 104 to 2.02 × 104 Da. Notably, the molar ratios of arabinose (Ara), galactose (Gal), and galacturonic acid (GalA) in LLPI decreased from 2.89 to 1.40, from 5.46 to 3.72, and from 21.24 to 18.71, respectively, suggesting that the utilization of arabinose and galactose in LLPI by intestinal microbiota was much faster than that of galacturonic acid at the initial fermentation stage. Additionally, LLPI could remarkably regulate gut microbial composition by increasing the abundances of several beneficial microbes, including Bacteroides, Bifidobacterium, Megamonas, and Collinsella, resulting in the promoted generation of several short-chain fatty acids, especially acetic, propionic, and butyric acids. The findings from the present study are beneficial to better understanding the digestive and microbial degradation characteristics of LLP, which indicate that LLP can be used as a potential prebiotic for the improvement of intestinal health.

Dynamic variations in physicochemical characteristics of oolong tea polysaccharides during simulated digestion and fecal fermentation in vitro.

In this study, dynamic variations in physicochemical characteristics of polysaccharides from 'Wuyi rock' tea (WYP) at different simulated digestion and fecal fermentation stages in vitro were studied. Results revealed that physicochemical characteristics of WYP were slightly altered after the simulated digestion in vitro, and its digestibility was about 8.38%. Conversely, physicochemical characteristics of the indigestible WYP, including reducing sugar, chemical composition, constituent monosaccharide, molecular weight, and FT-IR spectrum, were obviously altered after the fecal fermentation in vitro, and its fermentability was about 42.18%. Notably, the indigestible WYP could remarkably modulate the microbial composition via promoting the proliferation of profitable intestinal microbes, such as Bacteroides, Lactococcus, and Bifidobacterium. Moreover, it could also enhance the generation of short-chain fatty acids. The results showed that WYP was slightly digested in the gastrointestinal tract in vitro, but could be obviously utilized by intestinal microbiota, and might possess the potential to improve intestinal health.

Hydroxysafflor Yellow A Blocks HIF-1α Induction of NOX2 and Protects ZO-1 Protein in Cerebral Microvascular Endothelium.

Zonula occludens-1 (ZO-1) is a tight junction protein in the cerebrovascular endothelium, responsible for blood-brain barrier function. Hydroxysafflor yellow A (HSYA) is a major ingredient of safflower (Carthamus tinctorius L.) with antioxidative activity. This study investigated whether HSYA protected ZO-1 by targeting ROS-generating NADPH oxidases (NOXs). HSYA administration reduced cerebral vascular leakage with ZO-1 protection in mice after photothrombotic stroke, largely due to suppression of ROS-associated inflammation. In LPS-stimulated brain microvascular endothelial cells, HSYA increased the ratio of NAD+/NADH to restore Sirt1 induction, which bound to Von Hippel-Lindau to promote HIF-1αdegradation. NOX2 was the predominant isoform of NOXs in endothelial cells and HIF-1α transcriptionally upregulated p47phox and Nox2 subunits for the assembly of the NOX2 complex, but the signaling cascades were blocked by HSYA via HIF-1α inactivation. When oxidate stress impaired ZO-1 protein, HSYA attenuated carbonyl modification and prevented ZO-1 protein from 20S proteasomal degradation, eventually protecting endothelial integrity. In microvascular ZO-1 deficient mice, we further confirmed that HSYA protected cerebrovascular integrity and attenuated ischemic injury in a manner that was dependent on ZO-1 protection. HSYA blocked HIF-1α/NOX2 signaling cascades to protect ZO-1 stability, suggestive of a potential therapeutic strategy against ischemic brain injury.

Expression profiles of glutathione S-transferase genes in larval midgut of Bombyx mori exposed to insect hormones.

Glutathione S-transferases (GSTs) are believed to play a role in the detoxification of xenobiotics, resistance to insect viruses and pesticides, intracellular transport, biosynthesis of hormones and protection against oxidative stress. In this study, we used quantitative real time RT-PCR to examine expression profiles of the silkworm Bombyx mori GST-Sigma (BmGSTS2) and GST-Delta (BmGSTD2) genes in the larval midgut of the silkworm after exposure to 2-hydroxyecdysone (20E) and juvenile hormone analog (JHA). In concentration-course study, 20E at higher concentrations (1.0 and 2.0 µg/µl) caused significant upregulation of BmGSTD2, and all concentrations (0.5-2.0 µg/µl) of 20E caused significant upregulation of BmGSTS2. However, JHA in all concentrations downregulated the expression of BmGSTD2 and BmGSTS2. When exposed to either 20E (2.0 µg/µl) or JHA (2.0 µg/µl) on the third day of the fifth instar, the silkworm had higher BmGSTD2 at later time points: 15, 18, and 24 h for 20E and 24 h for JHA. BmGSTS2 expression was downregulated within 24 h after exposure to JHA and showed a time-dependent response after exposure to 20E. We also did a stage-dependent study, in which JHA downregulated BmGSTD2 expression and upregulated BmGSTS2 expression significantly at both day 1 and day 3 of the fifth instar. 20E upregulated the expression of BmGSTD2 and BmGSTS2 at the two stages. These findings imply that hormones have an important role in the regulation of basal GST expression. However, further validation and field trials should be carried out on the regulatory elements relevant to BmGSTD2 and BmGSTS2 gene expression.

The discovery approaches and detection methods of microRNAs.

MicroRNAs (miRNAs) are small, highly conserved, non-coding RNAs that regulate gene expression of target mRNAs through cleavage or translational inhibition. Computer-based approaches for miRNA gene identification are being considered as indispensable in miRNAs research. Similarly, experimental approaches for detection of miRNAs are crucial to the testing and validating of computational algorithms. The detection of miRNAs in tissues or cells can supply valuable information for investigating the biological function of these molecules. Selective and highly sensitive detection methods will pave the way for extended understanding of miRNA function within organisms. In this review, we summarize the various computational methods for identification of miRNAs as well as the methodologies that have been developed to detection miRNAs.

Differential expression of microRNA-2b with potential target coding P25 in the fifth instar larvae posterior silk gland of the silkworm.

MicroRNAs (miRNAs) are a class of non-protein coding small RNAs that regulate a gene expression at the post-transcriptional level. Using in silico screening, we found that the 3'-untranslated regions of the P25 gene mRNA are perfectly complementary to nucleotides 2-8 at the 5' end of the miRNA-2b (miR-2b). The expression of miR-2b and the P25 gene in posterior silk gland of the fifth instar larval silkworm was investigated using real-time PCR detection method. The results indicated that expression of the P25 gene was very high in the posterior silk gland during the fifth instar larvae, whereas a level of miR-2b sharply decreased until reaching the lowest one on the 8th day. The expression patterns of miR-2b and P25 gene indicate that miR-2b might act as a fine-tuning regulator of expression of the P25 gene at the post-transcriptional level.

[Identification of Pu'er teas with different storage years by FTIR spectroscopy].

Infrared spectra of Pu'er raw tea and Pu'er ripe tea were investigated using Fourier transform spectroscopy, in order to exploit a rapid method for discrimination of aging period for Pu' er tea samples. The results showed that the two kinds of Pu'er teas shared a similar woveform of infrared spectrum. However, due to the variations of aging time, leading to different chemical composition in pu'er teas, both Pu'er raw tea and ripe tea displayed corresponding different characteristic peaks. And the extent of aging of Pu'er tea had a significant relationship with optical density and waveforms of absorption peaks in the wave number range of 1 120-1 570 cm(-1) and 400-853 cm(-1), suggesting that the extent of aging of Pu'er tea may be identified by infrared spectrum technology rapidly and simply.

HIF-1: structure, biology and natural modulators.

Hypoxia-inducible factor 1 (HIF-1), as a main transcriptional regulator of metabolic adaptation to changes in the oxygen environment, participates in many physiological and pathological processes in the body, and is closely related to the pathogenesis of many diseases. This review outlines the mechanisms of HIF-1 activation, its signaling pathways, natural inhibitors, and its roles in diseases. This article can provide new insights in the diagnosis and treatment of human diseases, and recent progress on the development of HIF-1 inhibitors.

Dynamic changes of structural characteristics of snow chrysanthemum polysaccharides during in vitro digestion and fecal fermentation and related impacts on gut microbiota.

The in vitro simulated saliva-gastrointestinal digestion and human fecal fermentation of snow chrysanthemum polysaccharides (JHP) were investigated. Results showed that reducing sugar contents of JHP increased during the gastrointestinal digestion, and glucose released with the decrease of its molecular weight, suggesting that JHP could be partially degraded under the gastrointestinal digestion. Furthermore, after in vitro fecal fermentation, the molecular weight and molar ratio of constituent monosaccharides (galactose and galacturonic acid) of the indigestible JHP (JHP-I) significantly decreased, and both monosaccharides and oligosaccharides released, suggesting that JHP-I could be further degraded and consumed by gut microbiota. Some beneficial bacteria, such as genera Bifidobacterium, Lactobacillus, Megamonas, and Megasphaera, significantly increased, suggesting that JHP-I could change the composition and abundance of gut microbiota. These results suggest that JHP is a potential source of prebiotics, and can be helpful for better understanding of the potential digestion and fermentation mechanism of JHP.

In vitro simulated digestion and fecal fermentation of polysaccharides from loquat leaves: Dynamic changes in physicochemical properties and impacts on human gut microbiota.

The aim of this study was to well understand the dynamic changes of physicochemical properties of polysaccharides from loquat leaves (LLP) during in vitro simulated saliva-gastrointestinal digestion and fecal fermentation and its related impacts on human gut microbiota. Results showed that the contents of reducing sugar of LLP slightly increased during the gastrointestinal digestion, and its molecular weight also slightly decreased, suggesting that LLP could be slightly degraded under the gastrointestinal digestion conditions. Moreover, during the fecal fermentation, the molecular weight of the indigestible LLP (LLP-I) significantly decreased, and the molar ratio of constituent monosaccharides of LLP-I, such as glucuronic acid, galacturonic acid, galactose, and arabinose, significantly changed, indicating that LLP-I could be degraded and consumed by human gut microbiota. Indeed, some beneficial bacteria such as Megasphaera, Megamonas, Bifidobacterium, Phascolarctobacterium, and Desulfovibrio significantly increased, suggesting that LLP-I could change the composition and abundance of gut microbiota. LLP-I could also promote the production of health-promoting short chain fatty acids. Results from this study are benefical to well understand the in vitro digestion and fecal fermentation behaviors of LLP, and LLP can be developed as a potential prebiotic in the functional food industry.

Polysaccharides from dandelion (Taraxacum mongolicum) leaves: Insights into innovative drying techniques on their structural characteristics and biological activities.

The aim of this study was to well understand the impacts of innovative drying techniques (radio frequency drying and microwave drying) and traditional drying techniques (vacuum drying, freezing drying, and hot air drying) on the structural characteristics and bioactivities of polysaccharides from dandelion leaves (DLPs). Five different DLPs were obtained from dandelion leaves dried by abovementioned drying techniques. Results showed that the structural characteristics and bioactivities of DLPs varied with different drying techniques. The molecular weights, apparent viscosities, molar ratios of constituent monosaccharide, contents of uronic acids, and contents of bonded polyphenolics in DLPs obtained by different drying techniques had noticeable variations, while the types of constituent monosaccharides and the major glycosidic linkages in DLPs were similar. In addition, results showed that DLPs, especially DLP-RF obtained by the radio frequency drying, exhibited remarkable antioxidant activities (ABTS, DPPH, and NO radical scavenging activities), excellent in vitro antiglycation activity, and obvious in vitro inhibitory activity on α-glucosidase. Results from this study suggest that the radio frequency drying can be used as a potential drying technique before extracting DLPs for applications in the functional food and medicine industries.

Involvement of volume-sensitive Cl- channels in the proliferation of human subcutaneous pre-adipocytes.

1. Obesity is a significant challenge in terms of public health and preventive medicine. Inhibition of pre-adipocyte proliferation is believed to be important in the proposed anti-obesity mechanism. The aim of the present study was to examine the interplay between Cl(-) channels and their possible involvement in the proliferation of undifferentiated human pre-adipocytes. 2. Pre-adipocytes were isolated from human abdominal subcutaneous adipose tissue. Membrane ion currents were recorded using the whole-cell patch-clamp technique. Expression of the Cl(-) channel ClC-3 gene and protein was determined by reverse transcription-polymerase chain reaction (RT-PCR) and western blot, respectively. Cell proliferation was evaluated using the [(3)H]-thymidine incorporation assay. 3. Electrophysiological recordings revealed a volume-sensitive Cl(-) current (I(Cl.vol)) expressed in pre-adipocytes that was activated under hyposmotic conditions (external osmolarity decreased by 80%) and inhibited by the Cl(-) channel blocker tamoxifen. 4. Expression of the ClC-3 channel gene and protein was confirmed by RT-PCR and western blot analysis. Blocking I(Cl.vol) with tamoxifen supressed the proliferation of pre-adipocytes in a concentration-dependent manner. 5. Collectively, the results of the present study indicate that the volume-sensitive Cl(-) channel participates in regulation of the proliferation of human subcutaneous pre-adipocytes.

Exercise benefits cardiovascular health in hyperlipidemia rats correlating with changes of the cardiac vagus nerve.

The role of exercise training on hemodynamic parameters, blood lipid profiles, inflammatory cytokines, cholinesterase-positive nerves and muscarinic cholinergic (M(2)) receptors expression in the heart was investigated in Sprague-Dawley male rats with hyperlipidemia (HL). The rats were subjected to a high-fat diet and exercise training for 8 weeks, and then the hemodynamic parameters, the profiles of blood lipid and inflammatory cytokines, and the expression of cholinesterase-positive nerves and M(2) receptors were measured. HL rats displayed cardiac dysfunction, dysregulation of inflammatory cytokines, and decreased cholinesterase-positive nerves and M(2) receptors expression. The combination of hyperlipidemia with exercise training (AT) restored the profiles of blood lipids and the levels of inflammatory cytokines. In addition, AT and HL + AT improved cardiac function with increasing cholinesterase-positive nerves and M(2) receptors expression. Overall, these data show that the increased expression of cholinesterase-positive nerves and M(2) receptors in the heart is partially responsible for the benefits of exercise training on cardiac function in hyperlipidemia rats.