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Eleven new nitrogenous meroterpenoids, cinerols A-K (1-11), were isolated from the marine sponge Dysidea cinerea collected in the South China Sea, and their structures were determined by detailed spectroscopic analysis. Cinerols A (1) and B (2) feature a rare 5H-pyrrolo[1,2a]benzimidazole moiety, while cinerols C-G (3-7) are examples of rare meroterpene benzoxazoles. The cinerols are noncytotoxic to human melanoma A375 cells at the concentration of 32 μM; however, selected cinerols exhibit moderate inhibitory activity against one or more of protein-tyrosine phosphatase 1B, ATP-citrate lyase, and SH2 domain-containing phosphatase-1 with IC50 values of 2.8-27 μM.

Uncoupling of mitochondrial respiration by chemical uncouplers has proven effective in ameliorating obesity, insulin resistance, and hyperglycemia. However, development of uncoupler-based therapy remains challenging due to its potentially lethal adverse effects. Here, we identify pyruvate dehydrogenase (PDH) as a key modifier of the toxicity profile of 2, 4-dinitrophenol (DNP), a prototypical mitochondrial uncoupler. PDH activation by dichloroacetic acid (DCA) protects mice from DNP-induced hyperlactacidemia, hyperthermia, and death while preserving the ability of DNP to promote fuel oxidation and improve insulin sensitivity in mice. Mechanistically, PDH activation switches on mitochondrial glucose oxidation to accommodate increased glycolytic flux, leading to reduced lactate secretion during uncoupler treatments. We devised a chemical screening strategy and discovered compound 6j as a dual-action compound that simultaneously activates PDH and uncouples mitochondrial respiration. Compound 6j exhibits an excellent efficacy and safety profile in restoring glucose homeostasis in diabetic mice. This work establishes a new principle to safely harness the power of chemical uncouplers for the treatment of metabolic disease.

The chemical investigation of the marine sponge Dysidea frondosa discovered a pair of unprecedented bioconjugates that are composed of a meroterpene and an unusual psammaplysin alkaloid. The structures of frondoplysins A (1) and B (2) were characterized by analysis of HRMS and NMR data coupled with single-crystal X-ray diffraction. Frondoplysin A was found to be a potent inhibitor targeting protein-tyrosine phosphatase 1B (PTP1B) with an IC50 value of 0.39 μM.

Several series of novel tryptophan-derived rhodanine derivatives were synthesized and identified as potential competitive PTP1B inhibitors and antibacterial agents. Among the compounds studied, 10b was found to have the best in vitro inhibition activity against PTP1B (IC50 = 0.36 ± 0.02 μM). In addition, the compounds also showed potent inhibition against other PTPs, especially CDC25B. Molecular docking analysis demonstrated that compounds 7c and 10b could occupy both the catalytic site and the adjacent pTyr binding site simultaneously. The compounds also showed higher levels of activity against gram-positive strains, the gram-negative strain Escherichia coli 1924, and multidrug-resistant gram-positive bacterial strains. Compounds 7c, 8c, 9e, 10a, and 10c had comparable or more potent antibacterial activity than the positive controls.

The unique thermogenic capacity of brown adipocyte makes it an attractive target for antiobesity treatments. Several epigenetic regulators can control brown adipocyte development. In this study, we show that SIRT5, a member of the sirtuins, is required for brown adipocyte differentiation and essential for brown adipogenic gene activation in vitro. Furthermore, we find out that knockdown of SIRT5 reduces intracellular α-ketoglutarate concentration, which leads to elevated H3K9me2 and H3K9me3 levels at promoter regions of Pparγ and Prdm16 loci. Finally, we discover that SIRT5 knockout mice on the Sv129 background exhibit less browning capacity in subcutaneous white adipose tissue compared with controls and show apparent cold intolerance, suggesting that SIRT5 can modulate the browning process in vivo. Thus, our study uncovers a new biological function of SIRT5 in brown adipocyte differentiation and a mechanism by which SIRT5 regulates brown adipogenic gene activation at least partly through an indirect effect on histone modifications. Our study extends the linkage between epigenetics and cell differentiation.

Four indole alkaloids, alstonlarsines A–D (1–4), were isolated from Alstonia scholaris and structurally characterized. Compound 1 possesses a new carbon skeleton with a cage-shaped 9-azatricyclo[4.3.1.03,8]decane motif, and compounds 2–4 feature a rare carbon skeleton that was found in nature for the first time. Plausible biosynthetic routes for 1–4 are proposed. Compound 1 showed DRAK2 inhibitory activity with an IC50 value of 11.65 ± 0.63 μΜ.

Insulin-induced gene (Insig) negatively regulates SREBP-mediated de novo fatty acid synthesis in the liver. However, the upstream regulation of Insig is incompletely understood. Here we report that AMPK interacts with and mediates phosphorylation of Insig. Thr222 phosphorylation following AMPK activation is required for protein stabilization of Insig-1, inhibition of cleavage and processing of SREBP-1, and lipogenic gene expression in response to metformin or A769662. AMPK-dependent phosphorylation ablates Insig’s interaction with E3 ubiquitin ligase gp78 and represses its ubiquitination and degradation, whereas AMPK deficiency shows opposite effects. Interestingly, activation of AMPK by metformin causes an augmentation of Insig stability and reduction of lipogenic gene expression, and leads to the attenuation of hepatic steatosis in HFHS diet-fed mice. Moreover, hepatic overexpression of Insig-1 rescues hepatic steatosis in liver-specific AMPKα2 knockout mice fed with HFHS diet. These findings uncover a novel effector of AMPK. Targeting Insig may have the therapeutic potential for treating fatty liver disease and related disorders., Insulin-related gene (Insig) negatively regulates hepatic fatty acid synthesis, a process involved in development of non-alcoholic fatty liver disease (NAFLD). Here, the authors show that AMPK activation by metformin promotes Insig phosphorylation, stabilizing it and inhibiting lipogenic gene expression. This is protective against steatosis in diabetic mice.

Poor medication adherence is one of the leading causes of suboptimal glycaemic control in approximately half of the patients with type 2 diabetes mellitus (T2DM). Long-acting antidiabetic drugs are clinically needed for improving patients' compliance. Dipeptidyl peptidase-4 (DPP-4) inhibitors play an increasingly important role in the treatment of T2DM because of their favorable properties of weight neutrality and hypoglycemia avoidance. Herein, we report the successful discovery and scale-up synthesis of compound 5, a structurally novel, potent, and long-acting DPP-4 inhibitor for the once-weekly treatment of T2DM. Inhibitor 5 has fast-associating and slow-dissociating binding kinetics profiles as well as slow clearance rate and long terminal half-life pharmacokinetic properties. A single-dose oral administration of 5 (3 mg/kg) inhibited80% of DPP-4 activity for more than 7 days in diabetic mice. The long-term antidiabetic efficacies of 5 (10 mg/kg, qw) were better than those of the once-weekly trelagliptin and omarigliptin, especially in decreasing the hemoglobin A1c level.

The widespread usage of plastic products in human life has led to extensive exposure to plasticizers and resulted in serious health problems for humans, which has become a focus of toxicology research in the world. We aimed to explore the potential mechanism of liver insulin resistance induced by di(2-ethylhexyl) phthalate (DEHP) and working on a novel treatment to alleviate insulin resistance caused by excessive exposure to DEHP. For this purpose, in vivo and in vitro experiments were conducted, and the pivotal factors in the insulin signaling pathway were analyzed. In vivo study showed DEHP could lead to liver injury and insulin resistance. DEHP could break the balance of oxidative stress and cause accumulation of inflammatory factors. Genomics and proteomics experiment results revealed that DEHP could inhibit the mRNA and protein expression of insulin receptor, insulin receptor substrate, PI3K/Akt/mTOR, and glucose transporter 4. Nevertheless, the liver insulin resistance induced by DEHP could be reversed by Verapamil (thioredoxin interacting protein (TXNIP) inhibitor). Thus, we confirmed that DEHP caused insulin resistance by affecting the TXNIP in liver, further damaging the conduction of insulin signaling pathway. Therefore, adding Verapamil to the treatment of patients with insulin resistance due to plasticizers might be more effective.

Palmitate (PA) exposure induces stress conditions featuring ROS accumulation and upregulation of p62 expression, resulting in autophagic flux blockage and cell apoptosis. Sulfuretin (Sul) is a natural product isolated from Rhus verniciflua Stokes; the cytoprotective effect of Sul on human hepatic L02 cells and mouse primary hepatocytes under PA-induced stress conditions was investigated in this study. Sul induced mitophagy by activation of p-TBK1 and LC3 and produced a concomitant decline in p62 expression. Autophagosome formation and mitophagy were assessed by the sensitive dual fluorescence reporter mCherry-EGFP-LC3B, and mitochondrial fragmentation was analyzed using MitoTracker Deep Red FM. A preliminary structure-activity relationship (SAR) for Sul was also investigated, and the phenolic hydroxyl group was found to be pivotal for maintaining the cytoprotective bioactivity of Sul. Furthermore, experiments using flow cytometry and western blots revealed that Sul reversed the cytotoxic effect stimulated by the autophagy inhibitors 3-methyladenine (3-MA) and chloroquine (CQ), and its cytoprotective effect was almost eliminated when the autophagy-related 5 (Atg5) gene was knocked down. These studies suggest that, in addition to its antioxidative effects, Sul stimulates mitophagy and restores impaired autophagic flux, thus protecting hepatic cells from apoptosis, and that Sul has potential future medical applications for hepatoprotection.

A column photobioreactor with internal light column (ILC-CPBR) was compared with CPBRv (without internal light column but with the same culture volume), IC-CPBR (with internal column but without the internal light) and CPBRh (without internal light column but with the same height of culture solution) not only in hydrodynamics characteristics and light intensity, but also in microalgal growth and CO2 fixation rate. The result showed that mass transfer coefficient of ILC-CPBR was 24.3% and 21.5% higher than that of CPBRv and CPBRh in 0.1 vvm. The adding of internal light column enhanced light intensity of microalgal growth by increasing light/dark ratio. Based on mutual enhancement of mass transfer coefficient and light intensity, the biomass increased by 82.4% and 64.4% in ILC-CPBR compared with those in CPBRv and CPBRh, and the peak of CO2 biofixation rate increased by 47.7% and 26.2% in ILC-CPBR compared with those in CPBRv and CPBRh.

Three new (1a/1b and 2b) and five known flavans (2a, 6a/6b, 7, and 8), one new flavan glycoside (3), and two new (4 and 5) and six known 1,3-diphenylpropanes (9-14) were isolated from the twigs of Broussonetia kazinoki. Compounds 1, 2, and 6 are scalemic mixtures, and were resolved by chiral HPLC to provide 1a/1b, 2a/2b, and 6a/6b. The structures of these compounds were elucidated by extensive spectroscopic methods, including NMR, MS, and ECD analyses. Compounds 1, 2, 5, 8, 9, 11, and 12 showed in vitro inhibition of protein tyrosine phosphatase 1B (PTP1B).

Nine undescribed acylphloroglucinol derivatives, oblatones A−I, along with three known ones, were isolated from Syzygium oblatum. Their structures were determined on the basis of extensive spectroscopic analysis, including NMR and MS data interpretation. Oblatones A and B possess an alkylated chromanone scaffold featuring a hemiketal moiety. Oblatones C and D are the first acylphloroglucinol derivatives with an α,β-unsaturated ketone lipid chain. Some of the isolates showed inhibitory effects on ATP citrate lyase in vitro. The binding mode of oblatone A was predicted by molecular docking.

Nonalcoholic steatohepatitis (NASH) is an advanced stage of nonalcoholic fatty liver disease (NAFLD) with serious consequences that currently lacks approved pharmacological therapies. Recent studies suggest the close relationship between the pathogenesis of NAFLD and the dysregulation of RNA splicing machinery. Here, we reveal death-associated protein kinase-related apoptosis-inducing kinase-2 (DRAK2) is markedly upregulated in the livers of both NAFLD/NASH patients and NAFLD/NASH diet-fed mice. Hepatic deletion of DRAK2 suppresses the progression of hepatic steatosis to NASH. Comprehensive analyses of the phosphoproteome and transcriptome indicated a crucial role of DRAK2 in RNA splicing and identified the splicing factor SRSF6 as a direct binding protein of DRAK2. Further studies demonstrated that binding to DRAK2 inhibits SRSF6 phosphorylation by the SRSF kinase SRPK1 and regulates alternative splicing of mitochondrial function-related genes. In conclusion, our findings reveal an indispensable role of DRAK2 in NAFLD/NASH and offer a potential therapeutic target for this disease.

In an effort to identify novel inhibitors of nuclear factor kappa B (NF-κB), twenty five pyranochalcone derivatives were synthesized and evaluated for their in vitro activities against TNF-α induced NF-κB inhibition in HEK293T cells. Among all of these derivatives, several displaying the same acrylate moiety on the B ring exhibited potent inhibition, with IC50 values ranging from 0.29 to 10.46 μM. A functional study of the most potent of these compounds, designated 6b, revealed that it significantly suppressed the transcriptional expression of inflammatory factor IL-1β in lipopolysaccharide-induced RAW 264.7 macrophages, and also mildly inhibited CCL2, IL6 and TNF-α. In addition, compound 6b was found to inhibit IL-1β released in LPS-induced BMDM cells. This study demonstrates that the inhibitory effect of 6b on LPS-stimulated inflammatory mediator production in the mouse macrophage cell line RAW 264.7 correlates with the suppression of the NF-κB and MAPK signaling pathways.

Nonalcoholic hepatosteatosis (NASH) is an advanced stage of nonalcoholic fatty liver disease (NAFLD) with serious consequences that currently lacks approved pharmacological therapies. Recent studies suggest the close relationship between the pathogenesis of NAFLD and the dysregulation of RNA splicing machinery. Here, we revealed death-associated protein kinase-related apoptosis-inducing kinase-2(Drak2) is markedly upregulated in the livers of both patients and NAFLD/NASH diets-fed mice. Hepatic deletion of Drak2 suppresses the progression of hepatic steatosis to NASH. Comprehensive analyses of the phosphoproteome and transcriptome, indicated a crucial role of Drak2 in RNA splicing, and identified the splicing factor SRSF6 as a direct binding protein of Drak2. Further studies demonstrated that the binding of Drak2 inhibits phosphorylation of SRSF6 by the SRSF kinase(SRPK1) and regulates alternative splicing of mitochondrial function-related genes. In conclusion, our findings reveal an indispensable role of Drak2 in NAFLD/NASH and offer a potential therapeutic target for this disease.

Post-stroke depression (PSD) severely affects recovery in patients with intracerebral hemorrhage (ICH). Although hydrogen gas (HA rodent model of ICH in the basal ganglia was established using autologous blood injection (30 μL). Mice were treated with 2.9% HCompared with female mice, HEstrogen was responsible for increased H

Phateacid esters (PAEs), such as dibutyl phthalate (DBP), have been widely used and human exposure results into serious toxic effects; such as the development of fatty liver disease. In the present study, SD rat models for in vivo study (normal and fatty liver model group) and hepatocytes for in vitro study (normal and abnormal lipid metabolism model group) were established to determine the effects of DBP on liver function and discover the possible mechanisms. Meanwhile, the peroxisome proliferator activated receptor (PPARα) blocker, GW6471, with the Adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) activator, AICAR, were applied in vitro study to clarify the role of PPARα/SREBP-1c/FAS/GPAT/AMPK signal pathway in the process. Results suggested that DBP could activate PPARα signaling pathway and affected the protein expression of SREBP, FAS and GPAT to cause hyperlipidemia and abnormal liver function. DBP also could inhibit the phosphorylation and activation of AMPK to inhibit the decomposition and metabolism of lipids. Interestingly, the effects of DBP could be alleviated by GW6471 and AICAR. Our experimental results provide reliable evidence that DBP exposure could further induce liver lipid metabolism disorder and other hepatic toxicity through PPARα/SREBP-1c/FAS/GPAT/AMPK signal pathway.

Gypenosides, extracts of Gynostemma yixingense, have been traditionally prescribed to improve metabolic syndrome in Asian folk and local traditional medicine hospitals. However, the mechanism of its action remains unclarified. In this work, our results indicated that chronic administration of 2α-OH-protopanoxadiol (GP2), a metabolite of gypenosides in vivo, protected mice from high-fat diet-induced obesity and improved glucose tolerance by improving intestinal L-cell function. Mechanistically, GP2 treatment inhibited the enzymatic activity of bile salt hydrolase and modulated the proportions of the gut microbiota, which led to an increase in the accumulation of tauro-β-muricholic acid (TβMCA) in the intestine. TβMCA induced GLP-1 production and secretion by reducing the transcriptional activity of nuclear receptor farnesoid X receptor (FXR). Transplantation of GP2-remodelled fecal microbiota into antibiotic-treated mice also increased the intestinal TβMCA content and improved intestinal L-cell function. These findings demonstrate that GP2 ameliorates metabolic syndrome at least partly through the intestinal FXR/GLP-1 axis via gut microbiota remodelling and also suggest that GP2 may serve as a promising oral therapeutic agent for metabolic syndrome.

Beige adipocytes have been considered as a potential strategy in anti-obesity therapy because of its thermogenic capacity. AMP- activated protein kinase (AMPK) plays important roles in regulating adipose tissue function. C29 is a novel pyrazolone derivative with AMPK activity. In the current study, we investigated the role of C29 in the regulation of thermogenesis using differentiated adipocytes and diet-induced obese mice, and explored the mechanisms that might be involved in energy expenditure via adipocyte AMPK activation. We showed that treatment with C29 (2.5–10 μM) concentration-dependently increased thermogenesis in differentiated preadipocytes separated from inguinal white adipose tissue (iWAT), evidenced by increased expression levels of thermogenesis markers such as Ucp1, Pgc-1α, Dio2, Prdm16, Cox7a1, Cox8b, Elovl3 , and Cidea , fatty acid oxidation (FAO) genes including Cpt1a, Lcad and Pparα, as well as beige-selective genes such as Cd137, Tmem26, Slc27a1 , and Tbx1 . In high-fat diet (HFD)- fed mice, oral administration of C29 (30 mg·kg −1 ·day −1 ) for 9 weeks alleviated HFD-induced obesity, promoted energy expenditure and modulated iWAT browning. However, these effects were not observed in adipose-specific AMPKα1/α2 knockout (AKO) mice following C29 administration. Together, this study demonstrates that C29 regulates energy balance via adipocyte AMPK. Our findings show that the discovery of AMPK activators that specifically target adipose tissue may have therapeutic potential for treating obesity-related metabolic diseases.

Eight pairs of meroterpenoid enantiomers and four achiral meroterpenoids were isolated from Rhododendron anthopogonoides Maxim. Seventeen of them, named (+)-/(-)-anthoponoids A-G, (+)-daurichromene D, and anthoponoids H and I, are undescribed compounds with structural diversity. Their structures were characterized herein by a combined application of spectroscopic techniques, X-ray crystallographic analysis, ECD calculation, and the modified Mosher's method. (+)-/(-)-Anthoponoid A and anthoponoid I are the first Rhododendron meroterpenoids found to possess a hexahydroxanthene motif and a diterpene unit, respectively. Some isolates were identified as NF-κB pathway inhibitors, and (+)-anthoponoid E, (-)-anthoponoid G, and anthoponoid H showed suppressive effects on LPS-induced inflammatory responses in RAW 264.7 macrophages.

Fibroblast growth factor 1 (FGF1) demonstrated protection against nonalcoholic fatty liver disease (NAFLD) in type 2 diabetic (T2D) and obese mice by uncertain mechanism. This study investigated the therapeutic activity and mechanism of a non-mitagenic FGF1 variant (FGF1ΔHBS) against NAFLD. FGF1ΔHBS administration was effective in 9-month old db/db mice with established NAFLD; liver weight, lipid deposition and inflammation declined and liver function improved. FGF1ΔHBS reduced oxidative stress by nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) and elevation of antioxidant protein expression. FGF1ΔHBS also inhibited activity and/or expression of lipogenic genes, coincident with phosphorylation of AMP-activated protein kinase (AMPK) and its substrates. Mechanistic studies on palmitate exposed hepatic cells demonstrated that NAFLD-like oxidative damage and lipid accumulation could be reversed by FGF1ΔHBS. In palmitate treated hepatic cells, siRNA knockdown of Nrf2 abolished only FGF1ΔHBS anti-oxidative actions but not lipid metabolism improvement. In contrast, AMPK inhibition by pharmacological agents or siRNA abolished both FGF1ΔHBS benefits on oxidative stress and lipid metabolism that was FGF receptor 4 (FGFR4) dependent. Further support of these in vitro findings is that liver specific AMPK knockout abolished therapeutic effects of FGF1ΔHBS against high-fat-high-sugar diet-induced hepatic steatosis. These findings indicate that FGF1ΔHBS is effective in late stage NAFLD and acts by activation of AMPK via hepatocyte FGFR4. Disclosure Q. Lin: None. G. Cai: None. Z. Liu: None. J. Li: None. J. Li: None. K.A. Wintergerst: None. P.N. Epstein: None. L. Cai: None. Y. Li: None. Y. Tan: None. Funding American Diabetes Association (1-13-JF-53 to Y.T.), (1-18-IBS-082 to L.C.)

Low-protein/high-carbohydrate (LPHC) diet promotes metabolic health and longevity in adult humans and animal models. However, the complex molecular underpinnings of how LPHC diet leads to metabolic benefits remain elusive. Through a multi-layered approach, here we observed that LPHC diet promotes an energy-dissipating response consisting in the parallel recruitment of canonical and non-canonical (muscular) thermogenic systems in subcutaneous white adipose tissue (sWAT). In particular, we measured Ucp1 induction in association with up-regulation of actomyosin components and several Serca (Serca1, Serca2a, Serca2b) ATPases. In beige adipocytes, we observed that AMPK activation is responsible for transducing the amino acid lowering in an enhanced fat catabolism, which sustains both Ucp1- and Serca-dependent energy dissipation. Limiting AMPK activation counteracts the expression of brown fat and muscular genes, including Ucp1 and Serca, as well as mitochondrial oxidative genes. We observed that mitochondrial reactive oxygen species are the upstream molecules controlling AMPK-mediated metabolic rewiring in amino acid-restricted beige adipocytes. Our findings delineate a novel metabolic phenotype of responses to amino acid shortage, which recapitulates some of the benefits of cool temperature in sWAT. In conclusion, this highlights LPHC diet as a valuable and practicable strategy to prevent metabolic diseases through the enhancement of mitochondrial oxidative metabolism and the recruitment of different energy dissipating routes in beige adipocytes.

Hedyorienoids A (1) and B (2), two sesquiterpenoid dimers, were isolated and characterized from Hedyosmum orientale. Compound 1 was an unprecedented heterodimer of two different classes of sesquiterpenoids furnished by forming an unusual 1,3-dioxolane ring, while compound 2 possessed a new dimerization pattern of two guaiane-type sesquiterpenoids. Biosynthetic pathways for 1 and 2 were proposed with the coexisting monomers chloranthalactone A (3) and hedyosumin A (4) as the precursors. Compounds 2 and 3 showed significant NF-κB inhibitory activity.

Eight enantiomeric pairs of new meromonoterpenoids (1a/1b-8a/8b) and four known compounds (9-12) were isolated from Rhododendron nyingchiense. Their structures were established by spectroscopic methods, quantum chemical calculations, and X-ray crystallography. The enantiomeric pairs were acquired from scalemic mixtures by chiral-phase HPLC and showed diverse heterocyclic frameworks. Compounds 1a/1b possess a rare 6/7/5/5 heterocyclic system, and 2a/2b incorporate a new 6/6/3/5 heterocyclic system featuring a quinone motif. Compounds 3a/3b represent the first meroterpenoids with a 6/6/5 ring system from the Rhododendron genus. Putative biosynthetic pathways of these compounds are proposed. Compounds 1b, 2a-4a, 8a, 8b, and 11 exhibited weak inhibitory effects on PTP1B, with IC50 values ranging from 5.7 ± 0.5 to 61.0 ± 4.8 μM.

(+)- and (−)-Mangelonoids A and B, two pairs of novel macrocyclic diterpenoid enantiomers featuring an unprecedented bicyclo[9.3.1]pentadecane core and a rare bridgehead double bond, were isolated from Croton mangelong. Their structures were unambiguously established using spectroscopic data, X-ray crystallography, and electronic circular dichroism analysis. A plausible biosynthesis pathway for these compounds was proposed. (−)-Mangelonoid A exhibited NF-κB inhibition with an IC50 value of 7.27 ± 1.30 μM.

Virus-like particles encapsulating HBV-RNA represent a serum biomarker for assessing viral replication activity in clinical practice. However, baseline levels of serum HBV-RNA and their associations with viral replicative intermediates and liver disease in phases of chronic hepatitis B remain unknown. In this cross-sectional study, 102 patients were categorized into immune-tolerant (IT), HBeAg-positive immune active (HBeAg+IA), inactive carrier (IC) and HBeAg-negative immune active (HBeAg-IA) phases. HBV-RNA in serum samples and in 66 paired liver biopsies were quantified and correlated with serum ALT levels, histopathological scores and the levels of other viral replicative intermediates. Mean levels of serum HBV-RNA differed among phases, with the highest levels among IT (6.78 ± 0.83 log10 copies mL-1 ) patients, followed by HBeAg+IA (5.73 ± 1.16 log10 copies mL-1 ), HBeAg-IA (4.52 ± 1.25 log10 copies mL-1 ) and IC (2.96 ± 0.40 log10 copies mL-1 ) patients. Serum HBV-RNA levels correlated with HBV DNA in all phases, although correlations with other viral replicative intermediates weakened or disappeared when cases were stratified into phases. Distinct compositions of viral products were found among phases: the ratio of HBsAg to serum HBV-RNA was highest in IC patients, while the ratio of serum HBV-RNA to intrahepatic HBV-RNA and the ratio of intrahepatic HBV-DNA to intrahepatic HBV-RNA were significantly higher in IT patients. In conclusion, baseline levels of HBV-RNA and the composition of viral replicative intermediates differ significantly across the natural course of chronic HBV infection. These findings shed light on the nature of viral replication and pathogenesis of disease among different phases of chronic HBV infection.

To discover more derivatives with better glucose-lowering efficacy compared with berberine, twenty-three novel compounds with 4,7,12,12a-tetrahydro-5H-thieno[3',2':3,4]pyrido[1,2-b]isoquinoline or 5,8,12,12a-tetrahydro-6H-thieno[2',3':4,5]pyrido[2,1-a]isoquinoline cores were designed, synthesized, and biologically evaluated in vitro in continuation of our previous work on indirect activators of adenosine 5'-monophosphate-activated protein kinase (AMPK). Nine compounds effectively stimulated glucose consumption (>2.3-fold at 10 μM) in L6 myotube cells, and two compounds (4d and 4s) exhibited superior inhibitory activity (

Forward osmosis (FO) has increasingly penetrated into traditional process, i.e. microalgae dewatering, to become one of the new energy saving technologies. However, FO membranes can be fouled by soluble algal products (SAP) and fouling behavior of the membrane is not clear due to the variety of SAP composition. In this work, two types of membranes including a self-made TFC and a commercial CTA, and three kinds of draw solutions including NaCl, MgCl2 and CaCl2 were adopted to investigate the dewatering of SAP from microalgae of Chlorella vulgaris. The dependence of permeate flux and foulants composition on the membranes, membrane orientation and reverse salt diffusion were then compared for membrane fouling behavior. The results showed that TFC membrane exhibited higher water permeability but more loss of water flux in comparison with CTA. The SAP was inclined to be adsorbed by TFC membrane while adsorbate was easier to be removed by physical cleaning. By contrast, the pollutants accumulated on the membrane surface of CTA were much more irreversible. The interaction between SAP from feed solution and calcium cations from draw solution induced the formation of cake layer on the surface of TFC membrane while irreversible granular deposits of SAP were found accumulated on CTA membrane surface. Possible membrane fouling mechanism was finally discussed for better fabrication of anti-fouling forward osmosis membranes, thus to propel the integration of forward osmosis with traditional field of microalgae dewatering.

Hongkonoids A-D (1-4), the first example of ascorbylated terpenoids featuring a unique 5,5,5-fused tricyclic spiroketal butyrolactone moiety and diterpenoid-derived long chain, were isolated from Dysoxylum hongkongense. Their structures were unambiguously assigned by a combination of spectroscopic data, chemical degradation, X-ray crystallography, CD analysis, and total synthesis. The total syntheses of compounds 1-4 were effectively accomplished by a convergent strategy with the longest linear sequences of 12-14 steps and overall yields of 5.4-9.6%. Notably, we exploited a bioinspired one-pot method to construct the key intermediate 14 from an easily made compound 12 by involving the cascade reactions of an elaborate Claisen rearrangement, deprotections, and a 5-exo-trig cyclization. The desired major epimer 14a was then transformed to the main building block 21. Assembly of 21 and the long chain vinyl iodide 7 was made by an NHK coupling reaction to furnish the framework of 1-4. Some of the hongkonoids and/or synthetic analogs showed significant to moderate inhibitory activities against NF-κB, 11β-HSD1, and sterol synthesis. The most active NF-κB inhibitor 34 exhibited distinct inhibition on the LPS-induced inflammatory responses in RAW 246.7 and primary BMDM cells.

Although Flotillin 1 (FLOT1) is highly expressed in various human cancers, its relationship with ovarian cancer (OC) remains unknown. This study determines FLOT1 expression in human ovarian tumors and examines its effect on OC cell proliferation. FLOT1 protein expression was assessed in a tissue microarray by immunohistochemical staining. We found that 81.48% malignant and 50% borderline tumors were FLOT1 protein-positive, whereas benign tumors and normal ovarian tissues were negative. The staining was strongest in serous malignant tumor and transitional cell carcinoma and weakest in mucinous tumor. Differentially expressed FLOT1 in freshly isolated serous tumors was confirmed by Western blot and we then evaluated FLOT1 expression association with OC patients' clinical characteristics. Histological typing established that FLOT1 protein expression was significantly associated with serous tumor (P Keywords: cell proliferation, epithelial ovarian cancer, FLOT1, miRNA, therapeutic target, tumorigenesis.

Due to chemoresistance and metastasis, the overall prognosis of osteosarcoma (OS) has not improved over the last two decades. Exploring novel therapeutic agents that can circumvent theses malignant phenotypes of OS would be essential to improve the survival of OS patients. Triptolide is a unique diterpene triepoxide that possesses potent antitumor activities.However, the effects and mechanism of triptolide on OS cells remain unknown. The effects of triptolide on viability, apoptosis, cell cycle distribution and migratory ability of OS cells were measured using MTT, flow cytometry and wound healing and transwell invasion assays. And an OS tumor xenograft mouse model was produced to further study the in vivo antitumor effects of triptolide. The expression of DUSP1 at the protein and mRNA level in OS cells was detected by western blot and qPCR. We report that triptolide exhibits multidimensional antitumor activities in OS cells, including the induction of apoptosis and G1 phase accumulation, inhibition of cell viability, migration, and invasion. We further demonstrate that triptolide inhibits the expression of dual-specificity protein phosphatase1 (DUSP1) through inhibiting its promoter activity, which causes sustained activation of three subfamilies of mitogen-activated protein kinase (MAPK). And the modulation of DUSP1/MAPK cascade is associated with the apoptosis of OS cells, since the ectopic expression of DUSP1 or the inhibition of MAPK using specific inhibitors can counteract triptolide-induced apoptosis. In addition, triptolide enhances doxorubicin-induced apoptosis. In summary, our study suggests that DUSP1 is an important cellular target of triptolide, and triptolide may be a promising treatment option for OS as a single agent or combined with other chemotherapeutics.

Two new Diels-Alder adducts, albasins A and B (1 and 2), one new isoprenylated 2-arylbenzofuran, albasin C (3), one new isoprenylated flavone, albasin D (4), together with sixteen known phenolic compounds, were isolated from the root bark of Morus alba. Their structures were elucidated by extensive spectroscopic analysis, including NMR, MS, and ECD data. All the new compounds and most of the known ones showed significant inhibitory effects on PTP1B in vitro with IC50 values ranging from 0.57 to 7.49μM.

The development of a selective and sensitive sensor for uric acid (UA) is of great significance in diagnosing disorders and avoiding relevant complications. Herein, a new colorimetric sensing platform for the detection of UA was established by Cu2+-catalyzed TMB (3,3′,5,5′-tetramethylbenzidine)–H2O2 system, in which H2O2 was produced from the catalytic reaction of UA by uricase. In this strategy, Cu2+ exhibited a highly efficient peroxidase-like activity and catalyzed the oxidation of TMB by H2O2 to produce a blue product, thus generating optical signals, which could be distinguished by naked eyes or detected through UV–vis spectrometer. The UA biosensor exhibited a linear response in the range of 1–100 μM, with a detection limit of 0.64 μM. The colorimetric sensor could discriminatively detect UA in human urine samples with the recoveries over 94.0%. The obtained results are consistent with those measured by biochemistry analyzer with the average relative error of 4.9%. The uricase involved and Cu2+-catalyzed TMB–H2O2 sensor offers the advantages of high selectivity and sensitivity, and no need of sample pretreatment. Therefore, the present UA biosensor has great potential in applications for biochemical analysis and clinical diagnosis. This strategy broadens the sensing application of Cu2+-catalyzed TMB–H2O2 system.

The present study was designed to develop a concise synthetic route for macrolide, with the purpose of confirming the absolute configuration of natural dihydroresorcylide (1) and making it more easily accessible for biological evaluation. The absolute configuration of C-3 in natural 1 was revised to be R by comparison of the rotation sign of synthetic (R)- and (S)-1. The synthetic (R)-1 was found to be a novel highly specific PTP1B inhibitor with an IC50 value of 17.06 μM.

Hepatocellular carcinoma (HCC) is one of the most difficult cancer disease for diagnosis and treatment, with a low survival rate and high recurrence rate and mortality. Nuclear factor of activated T-cells 5 (NFAT5) is mediated by osmolality and proved to be a carcinogenic gene in some tumor. However in our study we considered NFAT5 as tumor suppressor of HCC. RT-qPCR was performed for NFAT5 expression in tumor tissues. NaCl was applied to make hyperosmotic treatment. We knockdowned and overexpressed NFAT5 to investigate its role in HCC. FCM was used for apoptosis assay. Transwell and scratch assay is proceeded for invasion.NFAT5 is downregulated in HCC tissue and cell lines, besides, upregulated by hyperosmolality. NFAT5 promotes apoptosis by regulating PARP-1,BAX/BCL2 while inhibits invasion through EMT-related protein claudin-1 and fibronectin. Hyperosmolality is also a protective factor for HCC. We considered hyperosmolality exhibited his protective effect by inducing NFAT5.In a word, NFAT5 inhibits invasion and promotes apoptosis in HCC, associated with osmolality.

Macdentichalcone (1), an unprecedented polycyclic dimeric chalcone featuring a unique quinonoid moiety, was isolated from Macaranga denticulata, together with 1-(5,7-dihydroxy-2,2,6-trimethyl-2H-1-benzopyran-8-yl)-3-phenyl-2-propen-1-one (2), a known monomeric chalcone proposed as a biosynthetic precursor of 1. The structure of 1 was elucidated by extensive spectroscopic analysis including NMR and MS data, as well as calculation of 13C NMR chemical shifts. Both compounds 1 and 2 showed inhibitory activity against PTP1B in vitro.

Eleven new nitrogenous meroterpenoids, cinerols A-K (

Adenosine monophosphate-activated protein kinase (AMPK) has been considered as a promising drug target for its regulation in both glucose and lipid metabolism. Mogrol was originally identified from high throughput screening as a small molecule activator of AMPK subtype α2β1γ1. In order to enhance its potency on AMPK and summarize the structure-activity relationships, a series of mogrol derivatives were designed, synthesized and evaluated in pharmacological AMPK activation assays. The results showed that the amine derivatives at the 24-position can improve the potency. Among them, compounds 3 and 4 exhibited the best potency (EC50: 0.15 and 0.14 μM) which was 20 times more potent than mogrol (EC50: 3.0 μM).

Berberine (BBR) exhibits diverse bioactivities, including anticancer activity; but its poor druggability limits its applications. In this study, we designed and synthesized a series of 9-O position modified BBR derivatives aiming to improve its cell permeability and anticancer activity, utilizing a long alkyl chain branched by hydroxyl group and methoxycarbonyl group. Among these compounds, B10 showed 3.6-fold higher intracellular concentration than BBR, as well as 60-fold increased anti-proliferation activity against human lung cancer A549 cells compared with BBR. Treatment with B10 (1, 2 μM) induced apoptosis of A549 cells. Further investigations showed that B10 treatment dose-dependently affected mitochondrial functions, including oxygen consumption rate (OCR), mitochondrial membrane potential (MMP) and the morphology of mitochondria in A549 cells. Therefore, this work offers a new way for BBR structural modification through improving cell membrane permeability to affect mitochondrial functions and potential anti-tumor therapy in the future.

An extensive phytochemical investigation on the chemical constituents from the needles and twigs of the vulnerable conifer Pseudotsuga sinensis yielded 19 diterpenoids and 21 sesquiterpenoids with various carbocyclic skeletons. Among them, 13 (named pseudosinins A-M, resp.) were undescribed compounds. Their structures with absolute configurations were characterized by a combination of spectroscopic methods, calculated and experimental electronic circular dichroism (ECD) data, quantum chemical calculations of the chemical shifts, and single crystal X-ray diffraction analyses. In particular, an array of labdane-derived norditerpenoids with C

The chemical investigation of the marine sponge

Brown adipose tissue (BAT) dissipates metabolic energy and mediates non-shivering thermogenesis, thereby boosting energy expenditure. Increasing BAT mass and activity is expected to be a promising strategy for combating obesity; however, few medications effectively and safely recruit and activate BAT in humans. Berberine (BBR), a natural compound, is commonly used as a nonprescription drug to treat diarrhea. Here, we reported that 1-month BBR intervention increased BAT mass and activity, reduced body weight, and improved insulin sensitivity in mildly overweight patients with non-alcoholic fatty liver disease. Chronic BBR treatment promoted BAT development by stimulating the expression of brown adipogenic genes, enhanced BAT thermogenesis, and global energy expenditure in diet-induced obese mice and chow-fed lean mice, Consistently, BBR facilitated brown adipocyte differentiation in both mouse and human primary brown preadipocytes. We further found that BBR increased the transcription of PRDM16, a master regulator of brown/beige adipogenesis, by inducing the active DNA demethylation of PRDM16 promoter, which might be driven by the activation of AMPK and production of its downstream tricarboxylic acid cycle intermediate α-Ketoglutarate. Moreover, chronic BBR administration had no impact on the BAT thermogenesis in adipose-specific AMPKa1 and AMPKa2 knockout mice. In summary, we found that BBR intervention promoted recruitment and activation of BAT and AMPK–PRDM16 axis was indispensable for the pro-BAT and pro-energy expenditure properties of BBR. Our findings suggest that BBR may be a promising drug for obesity and related metabolic disorders in humans partially through activating BAT.

Type 2 diabetes (T2D), which is characterized by hyperglycemia and insulin resistance, has become global epidemic. Uncoupling of mitochondria by chemical uncoupler, 2,4-dinitrophenol (DNP), is an efficient way to ameliorate hyperglycemia and insulin resistance. However, DNP use ceased in 1930s due to the systemic toxicities including hyperlactacidemia, hyperthermia and even death, which were mainly resulted from uncoupling actions induced excessive fat combustion and anaerobic glycolysis. To relieve the adverse effects derived from uncoupling activity, here we identified an effect-enhancing and toxicity-reducing way linking activation of pyruvate dehydrogenase (PDH) with DNP to T2D therapy. Activation of PDH by its classical activator, dichloroacetic acid (DCA), protected mice from DNP-derived hyperlactacidemia, hyperthermia and even death. Meanwhile, DCA combined with DNP further promoted glucose disposal in streptozotocin-induced hyperglycemic mice and enhanced insulin signaling in high-fat diet mice. Mechanistically, PDH activation corrected the excessive fatty acid oxidation and anaerobic glycolysis by DNP that was strongly associated with the adverse effects, and preferentially promoted glucose oxidation to improve hyperglycemia. Based on the beneficial effects of DNP combined with DCA, a novel compound possessing both mitochondrial uncoupling and PDH activation effects, named 6j, was identified and confirmed to promote glucose oxidation preferentially in vitro and in vivo. Therefore, our study suggested that combination of mitochondrial uncoupling and PDH activation might be a novel effective pharmacological approach for safely treating T2D. Disclosure J. Li: None. J. Li: None. Funding National Natural Science Foundation of China; Chinese Academy of Sciences; Shanghai Science and Technology Commission; K.C. Wong Education Foundation

Islet apoptosis is a detrimental event causing or aggravating type 2 diabetes (T2D). The attenuation of islet β cells is always accompanied by the development of T2D. Current clinical antidiabetic drugs are shown to be effective, but they have not fundamentally solved the essence of islet cellular apoptosis and failure. Therefore, it is necessary to identify novel targets and potential therapeutics for hyperglycemic treatment with significant protection/delay islet function attenuation. DAP-related apoptosis-inducing kinase-2 (Drak2) is a member of death-associated protein kinase (DAPK) family. We found higher expression of Drak2 in pancreas of db/db mice compared with littermites. We demonstrated that Drak2 activity was negatively related with glucose-stimulated insulin secretion (GSIS) and the autophagic flux by knockdown or overexpression of Drak2 in primary islets. To further investigate the role of Drak2 in islet function, we generated β cell-specific Drak2 knockout mice (βKO) model. We found that the capacity of glucose tolerance was enhanced in βKO mice. And Drak2-deficient islets can resistant to palmitate-triggered apoptosis and GSIS impairment. On the other hand, Drak2 inhibitors were applied in the efficacy study, we demonstrated that these compounds effectively protected islets from apoptosis via autophagy promotion in vitro and ameliorated T2D in vivo in mouse models. Mechanistically, Drak2 may be involved in autophagy regulation through autophagy initiation complex, ULK1, to exerting its effects on the function and apoptosis of islet cells. Our preliminary work show that inhibition or deficiency of Drak2 protects β cell from apoptosis may due to autophagy promotion, and suggest that Drak2 might be a novel target for potential diabetic therapeutics development. Disclosure Y. lu: None. J. Li: None. J. Li: None. Funding Shanghai Municipal Science and Technology Commission (16430711700)