Myricitrin versus EGCG in the Treatment of Obesity: Target Mining and Molecular Mechanism Exploring based on Network Pharmacology.

BACKGROUND: Myricitrin is a flavonol glycoside possessing beneficial effects on obesity, a rising global health issue that affects millions of people worldwide. However, the involving target and mechanism remain unclear. OBJECTIVE: In the present study, the anti-obesity targets and molecular mechanisms of Myricitrin, along with another flavanol Epigallocatechin gallate (EGCG), were explored through network pharmacology, bioinformatics, and molecular docking. METHODS: The potential targets for Myricitrin and EGCG were obtained from Pharmmaper, SwissTargetPrediction, TargetNet, SEA, Super-PRED, TCMSP, and STICH databases. Meanwhile, DEG targets were retrieved from GEO datasets, and obesity targets were collected from DrugBank, TTD, DisGeNet, OMIM, GeneCards, PharmGKB, and CTD databases. GO and KEGG pathway enrichment analyses were conducted through Metascape online tool. Protein-protein interaction (PPI) networks were also constructed for compound, DEG, and obesity targets to screen the core targets through MCODE analysis. The further screened-out key targets were finally verified through the compound-target-pathway-disease network, mRNA expression level, target-organ correlation, and molecular docking analyses. RESULTS: In total, 538 and 660 targets were identified for Myricitrin and EGCG, respectively, and 725 DEG targets and 1880 obesity targets were retrieved. GO and KEGG analysis revealed that Myricitrin and EGCG targets were enriched in the pathways correlating with obesity, cancer, diabetes, and cardiovascular disease. Furthermore, the intersection core targets for Myricitrin and EGCG function mainly through the regulation of responses to hormones and involving pathways in cancer. Above all, androgen receptor (AR), cyclin D1 (CCND1), early growth response protein 1 (EGR1), and estrogen receptor (ERS1) were identified as key targets in the compound-target-pathway-disease network for both Myricitrin and EGCG, with significant different mRNA expression between weight loss and control groups. Target-organ correlation analysis exhibited that AR and CCND1 showed high expression in adipocytes. Molecular docking also revealed good binding abilities between Myricitrin and EGCG, and all four receptor proteins. CONCLUSION: The present research integrated network pharmacology and bioinformatics approach to reveal the key targets of Myricitrin and EGCG against obesity. The results provided novel insights into the molecular mechanism of Myricitrin and EGCG in obesity prevention and treatment and laid the foundations for the exploitation of flavonoid-containing herbal resources.

Graphitized Carbon-Coated Iron Fluoride Nanocavities for Enhanced Kinetics of Multielectron Cathode Conversion Reactions.

As for the conversion-type iron fluoride (FeF3) cathode material with multielectron reactions for lithium-ion batteries (LIBs), sluggish reaction kinetics and low electrical conductivity pose certain limitations for the long-lasting reversible conversion processes. Herein, the three-dimensional porous nitrogen-doped carbon matrix in situ anchoring FeF3 nanocavities coated by graphitized carbon (FeF3/GC) are rationally prepared. Through the Kirkendall effect, the low-temperature fluorination of NF3 enables the resultant hollow FeF3 nanoparticles to possess a large number of lithium storage cavities and outer graphitized carbon structure, further effectively buffering the expansion of volume. The FeF3/GC cathode delivers a superior discharge capacity of 504.2 mAh g-1 after 1200 cycles at 1000 mA g-1, with a capacity decay rate of only 0.01% per cycle. Even at a rate of 5000 mA g-1, the composite cathode still delivers a discharge capacity of 309.6 mAh g-1. Impressively, the existence of graphitized carbon and the short Li+ diffusion length ensure fast electron/ion transfer, which significantly enhances the conversion reaction kinetics. This study aims to provide a promising strategy for the efficiency enhancement of multielectron cathode conversion reactions for LIBs.

The synergized diagnostic value of VTQ with chemokine CXCL13 in lung tumors.

Virtual Touch Tissue Quantification (VTQ) offers several advantages in the diagnosis of various lung diseases. Chemokine expression levels, such as CXCL13, play a vital role in the occurrence and development of tumors and aid in the diagnosis process. The purpose of this study was to evaluate the combined value of VTQ and changes in CXCL13 expression levels for the diagnosis of lung tumors. A total of 60 patients with thoracic nodules and pleural effusion were included, with 30 of them having malignant pleural effusion (based on pathology) and the remaining 30 having benign thoracic nodules and pleural effusion. The relative expression level of CXCL13 was measured in the collected pleural effusions using Enzyme-Linked Immunosorbent Assay (ELISA). The relationship between CXCL13 expression levels and various clinical features was analyzed. A Receiver Operating Characteristic (ROC) curve analysis was conducted on the VTQ results and relative expression levels of CXCL13, and the areas under the curve, critical values, sensitivity, and specificity were calculated. Multivariate analysis incorporating multiple indicators was performed to determine the accuracy of lung tumor diagnosis. The results showed that the expression levels of CXCL13 and VTQ were significantly higher in the lung cancer group compared to the control group (P < 0.05). In the Non-Small Cell Lung Cancer (NSCLC) group, CXCL13 expression levels increased with later TNM staging and poorer tumor differentiation. The expression level of CXCL13 in adenocarcinoma was higher than that in squamous cell carcinoma. The ROC curve analysis revealed that CXCL13 had an area under the curve (AUC) of 0.74 (0.61, 0.86) with an optimal cut-off value of 777.82 pg/ml for diagnosing lung tumors. The ROC curve analysis of VTQ showed an AUC of 0.67 (0.53, 0.82) with a sensitivity of 60.0% and a specificity of 83.3%, and an optimal diagnostic cut-off of 3.33 m/s. The combination of CXCL13 and VTQ for diagnosing thoracic tumors had an AUC of 0.842 (0.74, 0.94), which was significantly higher than either factor alone. The results of the study demonstrate the strong potential of combining VTQ results with chemokine CXCL13 expression levels for lung tumor diagnosis. Additionally, the findings suggest that elevated relative expression of CXCL13 in cases of malignant pleural effusion caused by non-small cell lung cancer may indicate a poor prognosis. This provides promising potential for using CXCL13 as a screening tool and prognostic indicator for patients with advanced lung cancer complicated by malignant pleural effusion.

A machine learning model based on ultrasound image features to assess the risk of sentinel lymph node metastasis in breast cancer patients: Applications of scikit-learn and SHAP.

Background: This study aimed to determine an optimal machine learning (ML) model for evaluating the preoperative diagnostic value of ultrasound signs of breast cancer lesions for sentinel lymph node (SLN) status. Method: This study retrospectively analyzed the ultrasound images and postoperative pathological findings of lesions in 952 breast cancer patients. Firstly, the univariate analysis of the relationship between the ultrasonographic features of breast cancer morphological features and SLN metastasis. Then, based on the ultrasound signs of breast cancer lesions, we screened ten ML models: support vector machine (SVM), extreme gradient boosting (XGBoost), random forest (RF), linear discriminant analysis (LDA), logistic regression (LR), naive bayesian model (NB), k-nearest neighbors (KNN), multilayer perceptron (MLP), long short-term memory (LSTM), and convolutional neural network (CNN). The diagnostic performance of the model was evaluated using the area under the receiver operating characteristic (ROC) curve (AUC), Kappa value, accuracy, F1-score, sensitivity, and specificity. Then we constructed a clinical prediction model which was based on the ML algorithm with the best diagnostic performance. Finally, we used SHapley Additive exPlanation (SHAP) to visualize and analyze the diagnostic process of the ML model. Results: Of 952 patients with breast cancer, 394 (41.4%) had SLN metastasis, and 558 (58.6%) had no metastasis. Univariate analysis found that the shape, orientation, margin, posterior features, calculations, architectural distortion, duct changes and suspicious lymph node of breast cancer lesions in ultrasound signs were associated with SLN metastasis. Among the 10 ML algorithms, XGBoost had the best comprehensive diagnostic performance for SLN metastasis, with Average-AUC of 0.952, Average-Kappa of 0.763, and Average-Accuracy of 0.891. The AUC of the XGBoost model in the validation cohort was 0.916, the accuracy was 0.846, the sensitivity was 0.870, the specificity was 0.862, and the F1-score was 0.826. The diagnostic performance of the XGBoost model was significantly higher than that of experienced radiologists in some cases (P<0.001). Using SHAP to visualize the interpretation of the ML model screen, it was found that the ultrasonic detection of suspicious lymph nodes, microcalcifications in the primary tumor, burrs on the edge of the primary tumor, and distortion of the tissue structure around the lesion contributed greatly to the diagnostic performance of the XGBoost model. Conclusions: The XGBoost model based on the ultrasound signs of the primary breast tumor and its surrounding tissues and lymph nodes has a high diagnostic performance for predicting SLN metastasis. Visual explanation using SHAP made it an effective tool for guiding clinical courses preoperatively.

Hypertrophic preconditioning cardioprotection after myocardial ischaemia/reperfusion injury involves ALDH2-dependent metabolism modulation.

Brief episodes of ischaemia and reperfusion render the heart resistant to subsequent prolonged ischaemic insult, termed ischaemic preconditioning. Here, we hypothesized that transient non-ischaemic stress by hypertrophic stimulation would induce endogenous cardioprotective signalling and enhance cardiac resistance to subsequent ischaemic damage. Transient transverse aortic constriction (TAC) or Ang-Ⅱ treatment was performed for 3-7 days in male mice and then withdrawn for several days by either aortic debanding or discontinuing Ang-Ⅱ treatment, followed by subsequent exposure to regional myocardial ischaemia by in situ coronary artery ligation. Following ischaemia/reperfusion (I/R) injury, myocardial infarct size and apoptosis were markedly reduced and contractile function was significantly improved in the TAC preconditioning group compared with that in the control group. Similar results were observed in mice receiving Ang-Ⅱ infusion. Mechanistically, TAC preconditioning enhanced ALDH2 activity, promoted AMPK activation and improved mitochondrial energy metabolism by increasing myocardial OXPHOS complex expression, elevating the mitochondrial ATP content and improving viable myocardium glucose uptake. Moreover, TAC preconditioning significantly mitigated I/R-induced myocardial iNOS/gp91phox activation, inhibited endoplasmic reticulum stress and ameliorated mitochondrial impairment. Using a pharmacological approach to inhibit AMPK signalling in the presence or absence of preconditioning, we demonstrated AMPK-dependent protective mechanisms of TAC preconditioning against I/R injury. Furthermore, treatment with adenovirus-encoded ALDH2 partially emulated the actions of hypertrophic preconditioning, as evidenced by improved mitochondrial metabolism, inhibited oxidative stress-induced mitochondrial damage and attenuated cell death through an AMPK-dependent mechanism, whereas genetic ablation of ALDH2 abrogated the aforementioned actions of TAC preconditioning. The present study demonstrates that preconditioning with hypertrophic stress protects the heart from I/R injury via mechanisms that improve mitochondrial metabolism, reduce oxidative/nitrative stress and inhibit apoptosis. ALDH2 is obligatorily required for the development of cardiac hypertrophic preconditioning and acts as the mediator of this process.

Combination of Erythromycin and Curcumin Alleviates Staphylococcus aureus Induced Osteomyelitis in Rats.

Osteomyelitis is commonly caused by Staphylococcus aureus. Both erythromycin and curcumin can suppress S. aureus growth, but their roles in osteomyelitis are barely studied. We aim to explore the activities of erythromycin and curcumin against chronical osteomyelitis induced by methicillin-resistant S. aureus (MRSA). Chronicle implant-induced osteomyelitis was established by MRSA infection in male Wistar rats. Four weeks after bacterial inoculation, rats received no treatment, erythromycin monotherapy, curcumin monotherapy, or erythromycin plus curcumin twice daily for 2 weeks. Bacterial levels, bone infection status, inflammatory signals and side effects were evaluated. Rats tolerated all treatments well, with no death or side effects such as, diarrhea and weight loss. Two days after treatment completion, erythromycin monotherapy did not suppress bacterial growth and had no effect in bone infection, although it reduced serum pro-inflammatory cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-6. Curcumin monotherapy slightly suppressed bacterial growth, alleviated bone infection and reduced TNF-α and IL-6. Erythromycin and curcumin combined treatment markedly suppressed bacterial growth, substantially alleviated bone infection and reduced TNF-α and IL-6. Combination of erythromycin and curcumin lead a much stronger efficiency against MRSA induced osteomyelitis in rats than monotherapy. Our study suggests that erythromycin and curcumin could be a new combination for treating MRSA induced osteomyelitis.

Solanum Muricatum Promotes Osteogenic Differentiation of Rat Bone Marrow Stromal Cells.

Solanum muricatum (SM), also known as pepino, is known for its antioxidative and anti-inflammatory effects. The aim of this study was to evaluate the effects of SM extract in promoting osteogenic differentiation and regulating the Wnt and bone morphogenetic protein (BMP) signaling pathways. Ingredients of pepino were extracted and identified. SM extracts were used to treat rat bone marrow stromal cells (BMSCs), followed by evaluating alkaline phosphatase activities and mineralization levels. The mRNA levels of osteogenic biomarkers, including OPN and Collagen I, were also evaluated with real-time polymerase chain reaction. After treatment with SM extracts, the expressions of key proteins in the Wnt and BMP signaling pathways were assessed. DKK-1 and noggin, which are Wnt and BMP inhibitors, respectively, were added with SM extracts to investigate the role of Wnt and BMP pathways in the ameliorating effects of SM extract in osteogenesis. Treatment of BMSCs with SM extract promoted osteogenesis. Meanwhile, upregulations in the Wnt and BMP pathways were also observed. However, inhibiting both pathways compromised the effects of SM extract in promoting osteogenic differentiation. SM extract promotes osteogenic differentiation in BMSCs via promoting the Wnt and BMP signaling pathways.

ß-Carotene suppresses osteoclastogenesis and bone resorption by suppressing NF-κB signaling pathway.

AIMS: ß-Carotene is a natural anti-oxidant, which has been used for treatment of cancer and cardiovascular diseases. Recently, the ameliorating function of ß-carotene in osteoporosis has been implicated. However, the precise mechanism of ß-carotene in prevention and treatment of osteoporosis is largely unknown. In the present study, we aimed to elucidate how ß-carotene affects osteoclast formation and bone resorption. MAIN METHODS: Bone marrow-derived monocytes/-macrophages (BMM) were exposed to 0.05, 0.1, 0.2, 0.4 and 0.6µM ß-carotene, followed by evaluation of cell viability, lactate dehydrogenase (LDH) release, receptor activator of nuclear factor kappa B ligand (RANKL)-induced osteoclastogenesis and resorption pits formation. Key factors in nuclear factor kappa B (NF-ĸB) and mitogen-activated protein kinases (MAPK) pathways were evaluated with western blot after BMM cells were exposed to RANKL and ß-carotene. The effects of ß-carotene in nuclear factor of activated T-cells cytoplasmic 1 (NFATc1), c-Fos and cathepsin K (CTSK) expression were also evaluated. KEY FINDINGS: ß-Carotene significantly inhibited BMM viability and promoted LDH release at concentrations of 0.4 and 0.6µM. A decrease in RANKL-induced osteoclastogenesis and resorption was also observed after ß-carotene treatment. ß-Carotene attenuated the NF-ĸB pathway activation by RANKL, with no effect on MAPK pathway. ß-Carotene suppressed the upregulation of NFATc1 and c-Fos by RANKL. SIGNIFICANCE: We clarified the anti-osteoclastogenic role of ß-carotene, which is mediated by NF-κB signaling.

Flaxseed oil ameliorates alcoholic liver disease via anti-inflammation and modulating gut microbiota in mice.

BACKGROUND: Alcoholic liver disease (ALD) represents a chronic wide-spectrum of liver injury caused by consistently excessive alcohol intake. Few satisfactory advances have been made in management of ALD. Thus, novel and more practical treatment options are urgently needed. Flaxseed oil (FO) is rich in α-linolenic acid (ALA), a plant-derived n-3 polyunsaturated fatty acids (PUFAs). However, the impact of dietary FO on chronic alcohol consumption remains unknown. METHODS: In this study, we assessed possible effects of dietary FO on attenuation of ALD and associated mechanisms in mice. Firstly, mice were randomly allocated into four groups: pair-fed (PF) with corn oil (CO) group (PF/CO); alcohol-fed (AF) with CO group (AF/CO); PF with FO group (PF/FO); AF with FO group (AF/FO). Each group was fed modified Lieber-DeCarli liquid diets containing isocaloric maltose dextrin a control or alcohol with corn oil and flaxseed oil, respectively. After 6 weeks feeding, mice were euthanized and associated indications were investigated. RESULTS: Body weight (BW) was significantly elevated in AF/FO group compared with AF/CO group. Dietary FO reduced the abnormal elevated aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels in chronic ethanol consumption. Amelioration of these parameters as well as liver injury via HE staining in dietary FO supplementation in ALD demonstrated that dietary FO can effectively benefit for the protection against ALD. To further understand the underlying mechanisms, we investigated the inflammatory cytokine levels and gut microbiota. A series of inflammatory cytokines, including TNF-α, IL-1ß, IL-6 and IL-10, were determined. As a result, TNF-α, IL-1ß and IL-6 were decreased in AF/FO group compared with control group; IL-10 showed no significant alteration between AF/CO and AF/FO groups (p > 0.05). Sequencing and analysis of gut microbiota gene indicated that a reduction of Porphyromonadaceae and Parasutterella, as well as an increase in Firmicutes and Parabacteroides, were seen in AF group compared with PF control. Furthermore, dietary FO in ethanol consumption group induced a significant reduction in Proteobacteria and Porphyromonadaceae compared with AF/CO group. CONCLUSION: Dietary FO ameliorates alcoholic liver disease via anti-inflammation and modulating gut microbiota, thus can potentially serve as an inexpensive interventions for the prevention and treatment of ALD.

Curcumin protects human adipose-derived mesenchymal stem cells against oxidative stress-induced inhibition of osteogenesis.

The detrimental effects of oxidative stress on the skeletal system have been documented, and understanding the mechanisms is important to design a therapeutic strategy. As an antioxidant and anti-inflammatory agent, the active ingredient of turmeric curcumin has been used as medication for numerous complications including bone loss. However, it is unclear if curcumin could influence the osteogenic potential of mesenchymal stem cells (MSCs), particularly in oxidative injuries. Here we demonstrate that curcumin treatment protects cell death caused by hydrogen peroxide (H2O2) exposure in human adipose-derived MSCs in vitro. Importantly, curcumin is able to enhance the osteoblast differentiation of human adipose-derived MSCs that is inhibited by H2O2. Notably, both oxidative stress and the inhibition of Wnt/ß-catenin signaling are attenuated by curcumin treatment. These results suggest that curcumin can promote osteoblast differentiation of MSCs and protect the inhibitory effect elicited by oxidative injury. The findings support potential use of curcumin or related antioxidants in MSC-based bone regeneration for disease related with oxidative stress-induced bone loss.

MicroRNA-29 facilitates transplantation of bone marrow-derived mesenchymal stem cells to alleviate pelvic floor dysfunction by repressing elastin.

BACKGROUND: Pelvic floor dysfunction (PFD) is a condition affecting many women worldwide, with symptoms including stress urinary incontinence (SUI) and pelvic organ prolapse (POP). We have previously demonstrated stable elastin-expressing bone marrow-derived mesenchymal stem cells (BMSCs) attenuated PFD in rats, and aim to further study the effect of microRNA-29a-3p regulation on elastin expression and efficacy of BMSC transplantation therapy. METHODS: We inhibited endogenous microRNA-29a-3p in BMSCs and investigated its effect on elastin expression by RT-PCR and Western blot. MicroRNA-29-inhibited BMSCs were then transplanted into PFD rats, accompanied by sustained release of bFGF using formulated bFGF in poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NP), followed by evaluation of urodynamic tests. RESULTS: MicroRNA-29a-3p inhibition resulted in upregulated expression and secretion of elastin in in vitro culture of BMSCs. After co-injection with PLGA-loaded bFGF NP into the PFD rats in vivo, microRNA-29a-3p-inhibited BMSCs significantly improved the urodynamic test results. CONCLUSIONS: Our multidisciplinary study, combining microRNA biology, genetically engineered BMSCs, and nanoparticle technology, provides an excellent stem cell-based therapy for repairing connective tissues and treating PFD.

New strategy to rescue the inhibition of osteogenesis of human bone marrow-derived mesenchymal stem cells under oxidative stress: combination of vitamin C and graphene foams.

To rescue the oxidative stress induced inhibition of osteogenesis, vitamin C (VC) was chemically modified onto three-dimensional graphene foams (3D GFs), then their regulation on osteogenesis of human bone marrow-derived mesenchymal stem cells (BM-MSCs) was studied. Combined action of VC + GF significantly decreased H2O2-induced oxidative stress, and rescued H2O2-inhibited cell viability, differentiation and osteogenesis of BM-MSCs in vitro. Further studies revealed that Wnt pathway may be involved in this protection of osteogenesis. Furthermore, an in vivo mouse model of BM-MSCs transplantation showed that VC + GF remarkably rescued oxidative stress inhibited calcium content and bone formation. The combination of VC and GF exhibited more pronounced protective effects against oxidative stress induced inhibition of osteogenesis, compared to monotherapy of VC or GF. Our study proposed a new strategy in stem cell-based therapies for treating bone diseases.

The effects of different angiotensin II type 1 receptor blockers on the regulation of the ACE-AngII-AT1 and ACE2-Ang(1-7)-Mas axes in pressure overload-induced cardiac remodeling in male mice.

Angiotensin II (AngII) type 1 receptor blockers (ARBs) have been effectively used in hypertension and cardiac remodeling. However, the differences among them are still unclear. We designed this study to examine and compare the effects of several ARBs widely used in clinics, including Olmesartan, Candesartan, Telmisartan, Losartan, Valsartan and Irbesartan, on the ACE-AngII-AT1 axis and the ACE2-Ang(1-7)-Mas axis during the development of cardiac remodeling after pressure overload. Although all of the six ARBs, attenuated the development of cardiac hypertrophy and heart failure induced by transverse aortic constriction (TAC) for 2 or 4weeks in the wild-type mice evaluated by echocardiography and hemodynamic measurements, the degree of attenuation by Olmesartan, Candesartan and Losartan tended to be larger than that of the other three drugs tested. Additionally, the degree of downregulation of the ACE-AngII-AT1 axis and upregulation of the ACE2-Ang(1-7)-Mas axis was higher in response to Olmesartan, Candesartan and Losartan administration in vivo and in vitro. Moreover, in angiotensinogen-knockdown mice, TAC-induced cardiac hypertrophy and heart failure were inhibited by Olmesartan, Candesartan and Losartan but not by Telmisartan, Valsartan and Irbesartan administration. Furthermore, only Olmesartan and Candesartan could downregulate the ACE-AngII-AT1 axis and upregulate the ACE2-Ang(1-7)-Mas axis in vitro. Our data suggest that Olmesartan, Candesartan and Losartan could effectively inhibit pressure overload-induced cardiac remodeling even when with knockdown of Ang II, possibly through upregulation of the expression of the ACE2-Ang(1-7)-Mas axis and downregulation of the expression of the ACE-AngII-AT1 axis. In contrast, Telmisartan, Valsartan and Irbesartan only played a role in the presence of AngII, and Losartan had no effect in the presence of AngII in vitro.

TNF-α-induced NF-κB activation upregulates microRNA-150-3p and inhibits osteogenesis of mesenchymal stem cells by targeting ß-catenin.

Although systemic or local inflammation, commonly featured by cytokine activation, is implicated in patients with bone loss, the underlying mechanisms are still elusive. As microRNAs (miR), a class of small non-coding RNAs involved in essential physiological processes, have been found in bone cells, we aimed to investigate the role of miR for modulating osteogenesis in inflammatory milieu using human bone marrow mesenchymal stem cells (hBM-MSCs). Induced by proinflammatory cytokine TNF-α, miR-150-3p was identified as a key player in suppressing osteogenic differentiation through downregulating ß-catenin, a transcriptional co-activator promoting bone formation. TNF-α treatment increased the levels of miR-150-3p, which directly targeted the 3'-UTR of ß-catenin mRNA and in turn repressed its expression. In addition, we observed that miR-150-3p expression was increased by TNF-α via IKK-dependent NF-κB signalling. There are three putative NF-κB binding sites in the promoter region of miR-150, and we identified -686 region as the major NF-κB binding site for stimulation of miR-150 expression by TNF-α. Finally, the osteogenic differentiation of hBM-MSCs was inhibited by either miR-150-3p overexpression or TNF-α treatment, which was prevented by anti-miR-150-3p oligonucleotides. Taken together, our data suggested that miR-150-3p integrated inflammation signalling and osteogenic differentiation and may contribute to the inhibition effects of inflammation on bone formation, thus expanding the pathophysiological functions of microRNAs in bone diseases.

Nucleosome Assembly Protein 1-Like 1 (Nap1l1) Regulates the Proliferation of Murine Induced Pluripotent Stem Cells.

BACKGROUND/AIMS: To investigate whether nucleosome assembly protein 1-like 1 (Nap1l1) regulates the proliferation of induced pluripotent stem cells (iPSC) and the potential mechanisms. METHODS: Nap1l1-knockdown-iPSC and Nap1l1-overexpression-iPSC were constructed by transfection of lentiviral particles. The proliferation of iPSC was detected by MTT analysis, and cell cycle was analyzed by flow cytometry. RESULTS: Nap1l1 overexpression promoted iPSC proliferation and induced G2/M transition compared to their control iPSC while Nap1l1-knockdown-iPSC dramatically displayed the reduced proliferation and accumulated G2/M phase cells. Further analysis showed that Nap1l1 overexpression in iPSC increased the expression of cyclin B1, downregulated the expression of p21 and p27, while knockdown of Nap1l1 showed the opposite effects. In addition, overexpression of Nap1l1 promoted the phosphorylation of AKT and ERK in iPSC, while knockdown of Nap1l1 inhibited the effects. However, these effects displayed in Nap1l1-overexpression-iPSC were greatly suppressed by the inhibition of AKT or ERK signaling. CONCLUSIONS: The results indicate that Nap1l1 promotes the proliferation of iPSC attributable to G2/M transition caused by downregulation of p27 and p21, and upregulation of cyclin B1, the activation of AKT or ERK is involved in the process. The present study has revealed a novel molecular mechanism involved in the proliferation of iPSC.

ψ-Bufarenogin, a novel anti-tumor compound, suppresses liver cancer growth by inhibiting receptor tyrosine kinase-mediated signaling.

Resistance of hepatocellular carcinoma (HCC) to existing chemotherapeutic agents largely contributes to the poor prognosis of patients, and discovery of novel anti-HCC drug is in an urgent need. Herein we report ψ-Bufarenogin, a novel active compound that we isolated from the extract of toad skin, exhibited potent therapeutic effect in xenografted human hepatoma without notable side effects. In vitro, ψ-Bufarenogin suppressed HCC cells proliferation through impeding cell cycle progression, and it facilitated cell apoptosis by downregulating Mcl-1 expression. Moreover, ψ-Bufarenogin decreased the number of hepatoma stem cells through Sox2 depression and exhibited synergistic effect with conventional chemotherapeutics. Mechanistic study revealed that ψ-Bufarenogin impaired the activation of MEK/ERK pathway, which is essential in the proliferation of hepatoma cells. ψ-Bufarenogin notably suppressed PI3-K/Akt cascade, which was required in ψ-Bufarenogin-mediated reduction of Mcl-1 and Sox2. ψ-Bufarenogin inhibited the auto-phosphorylation and activation of epithelial growth factor receptor (EGFR) and hepatocyte growth factor receptor (c-Met), thereafter suppressed their primary downstream cascades Raf/MEK/ERK and PI3-K/Akt signaling. Taken together, ψ-Bufarenogin suppressed HCC growth via inhibiting, at least partially, receptor tyrosine kinases-regulated signaling, suggesting that ψ-Bufarenogin could be a novel lead compound for anti-HCC drug.

Combination Treatment With Antihypertensive Agents Enhances the Effect of Qiliqiangxin on Chronic Pressure Overload-induced Cardiac Hypertrophy and Remodeling in Male Mice.

We previously showed that Qiliqiangxin (QL) capsules could ameliorate cardiac hypertrophy and remodeling in a mouse model of pressure overload. Here, we compared the effects of QL alone with those of QL combined with the following 3 types of antihypertensive drugs on cardiac remodeling and dysfunction induced by pressure overload for 4 weeks in mice: an angiotensin II type 1 receptor (AT1-R) blocker (ARB), an angiotensin-converting enzyme inhibitor (ACEI), and a ß-adrenergic receptor (ß-AR) blocker (BB). Adult male mice (C57B/L6) were subjected to either transverse aortic constriction or sham operation for 4 weeks, and the drugs (or saline) were orally administered through gastric tubes. Cardiac function and remodeling were evaluated through echocardiography, catheterization, histology, and analysis of hypertrophic gene expression. Cardiomyocyte apoptosis and autophagy, AT1-R and ß1-AR expression, and cell proliferation-related molecules were also examined. Although pressure overload-induced cardiac remodeling and dysfunction, hypertrophic gene reprogramming, AT1-R and ß1-AR expression, and ERK phosphorylation were significantly attenuated by QL alone, QL + ARB, QL + ACEI, and QL + BB, the attenuation was stronger in the combination treatment groups. Moreover, apoptosis was reduced to a larger extent by each combination treatment than by QL alone, whereas autophagy was more strongly attenuated by either QL + ARB or QL + ACEI. None of the treatments significantly upregulated ErbB2 or ErbB4 phosphorylation, and none significantly downregulated C/EBPß expression. Therefore, the effects of QL on chronic pressure overload-induced cardiac remodeling may be significantly increased when QL is combined with an ARB, an ACEI, or a BB.

Method development and validation for pharmacokinetic and tissue distributions of ellagic acid using ultrahigh performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).

Ellagic acid is a dietary polyphenol found in numerous fruits and vegetables, possessing several health benefits such as antioxidant, anticancer and anti-atherosclerotic biological properties. The purpose of this study was to explore the pharmacokinetics and tissue distribution of ellagic acid in rats. A simple, rapid, sensitive and specific liquid chromatography-tandem mass spectrometry method to determine the ellagic acid in plasma and tissue samples was developed and validated. The separation was achieved using reversed-phase ultra-performance liquid chromatography (UPLC), and the mass spectrometric detection was achieved using heated electrospray ionization (negative mode) and multiple ion monitoring (m/z 301/229). A sample cleanup with a solid phase extraction (SPE) step prior to the UPLC-MS/MS analysis was also developed. The SPE and UPLC-MS/MS method established here was successfully applied to reveal the pharmacokinetic profiles and tissue distribution of ellagic acid. After oral administration dosing at 50 mg/kg, plasma levels of ellagic acid peaked at about 0.5 h, with Cmax value of 93.6 ng/mL, and the results showed that the ellagic acid was poorly absorbed after oral administration. The pharmacokinetic profile of ellagic acid fitted to a two-compartment model with t1/2α 0.25 h and t1/2ß 6.86 h, respectively. Following oral administration, ellagic acid was detected in all examined tissues including kidney, liver, heart, lung and brain et al., and the highest levels were found in kidney and liver.

Structural characterisation and antioxidant activity evaluation of phenolic compounds from cold-pressed Perilla frutescens var. arguta seed flour.

A total of 11 phenolic compounds, as well as sucrose (12) and tryptophan (13), were isolated from cold-pressed Perilla frutescens var. arguta seed flour using column chromatography, and their chemical structures were identified as 3'-dehydroxyl-rosmarinic acid-3-o-glucoside (1), rosmarinic acid-3-o-glucoside (2), rosmarinic acid (3), rosmarinic acid methyl ester (4), luteolin (5), luteolin-5-o-glucoside (6), apigenin (7), caffeic acid (8), caffeic acid-3-o-glucoside (9), vanillic acid (10) and cimidahurinine (11) using NMR and time-of-flight mass spectrometry. Of these components, compound 1 is novel, and this is the first report of compounds 10 and 11 in perilla seeds. HPLC quantification combined with antioxidant activity evaluation revealed that rosmarinic acid and rosmarinic acid-3-o-glucoside were the dominant phenolic antioxidants with strong antioxidant activities.

α-Glucosidase inhibitory activity of polyphenols from the burs of Castanea mollissima Blume.

Polyphenol extracts from the burs of Castanea mollissima Blume (CMPE) exhibited potential antioxidant and hypoglycemic activities. The α-glucosidase inhibitory activities of CMPE were assessed as a means of elucidating the mechanism behind its hypoglycemic activities. In vitro studies showed that CMPE significantly inhibited both yeast α-glucosidase, through a noncompetitive mode with an IC50 of 0.33 µg/mL, and rat intestinal α-glucosidase. In vivo studies revealed that oral administration of CMPE at doses of 600 mg/kg significantly reduced postprandial blood glucose levels by 27.2% in normal rats following sucrose challenges. Gel permeation chromatography revealed that CMPE exhibited typical characteristics of high-molecular-mass polymers with mean (Mn) and weight (Mw) average molecular weights of 35.4 and 50.7 kDa, respectively, and a polydispersity (Mw/Mn) of 1.432. Acid hydrolysis analysis indicated the presence of ellagitannins. These data suggest that CMPE, enriched with ellagitannins, would be an efficacious dietary supplement for diabetes management through the inhibition of alpha-glucosidase.