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We developed a new on-line method of ultra-performance liquid chromatography coupled with biochemical detection (UHPLC-BCD) to screen acetylcholinesterase (AChE) inhibitors in complex matrixes. Chromatography separation was performed using an Xtimate UHPLC C18 column (100 mm × 2.1 mm, 1.8 μm) and a gradient elution with methanol–0.1% formic acid at a flow rate of 0.08 mL/min. The BCD was based on a colorimetric method using Ellman’s reagent, and the detection wavelength was at 405 nm. Galanthamine was used as a positive reference to validate the methodology. The detection and quantitation limits of the UHPLC-BCD method were 0.018 and 0.060 μg, respectively. A functional equation was generated in terms of the negative peak area (X) and galanthamine concentration (Y, μg/mL). The regression equation was Y = 0.0028X2 + 0.4574X + 50.7776, R2 = 0.9993. UHPLC-fourier-transform mass spectrometry detection results revealed that five alkaloids showed obvious AChE inhibitory activities including coptisin, epiberberine, jatrorrhizine, berberine and palmatine. The relative AChE inhibitory activities of jatrorrhizine, berberine and palmatine in the Coptidis Rhizoma sample were equal to that of 257.0, 2355 and 283.9 μg/mL of galanthamine, respectively. This work demonstrated that the UHPLC-BCD method was convenient and feasible, and could be widely used for the screening and activity evaluation of the bioactive components in the complex extracts.

Objective To prepare a self-microemulsifying drug delivery system (SMEDDS) to increase the solubility and oral bioavailability of chrysin. Methods The preparation conditions were determined using factor analysis method. Preliminarily screening was conducted using compatibility tests and pseudo-ternary phase diagram studies. The central composite design-response surface methodology was used to determine the maximum drug loading and optimize SMEDDS formation, as characterized by surface morphology, pH, diameter, polydispersity index (PDI), zeta potential, and phase type. In vitro release of chrysin-suspension and chrysin-SMEDDS was investigated using the bulk-equilibrium reverse dialysis bag technique. Short-term stability of chrysin-SMEDDS at high and low temperatures was assessed. Pharmacokinetic behaviors were evaluated after intragastric and intravenous administration to rats. Results The final optimal formulation was medium chain triglyceride:oleic acid:Cremophor RH40: Transcutol HP (w/w) (12%:12%:32%:44%), with a drug loading capacity of 5 mg/g. Diluted chrysin-SMEDDS was characterized as an oil-in-water type and spherical, with a diameter, pH, PDI, and zeta potential of 28.26 ± 0.83 nm, 5.60 ± 0.84, 0.18 ± 0.01, and -23.13 ± 0.95 mV, respectively. The release speed of chrysin-SMEDDS was significantly higher than that of chrysin-suspension, and the release process was not affected by the media pH. In vivo pharmacokinetic data revealed that the oral bioavailability of chrysin-SMEDDS was 2.7-fold higher than that of chrysin suspension, compared with the chrysin microemulsion. Conclusion The optimal SMEDDS formulation increased the dissolution and oral bioavailability of chrysin and may be useful for investigating chrysin efficacy in animal disease models and toxicokinetic studies.

Lupeol liposomes, modified with Gal-PEG-DSPE, were developed following a thin-film dispersion method. Then, the morphology, physicochemical properties, and in vitro release properties of those liposomes were investigated. The scanning electron microscopic images showed that most of the liposomes were spherical particles; they were similar in size and uniformly dispersed. Both lupeol liposomes and Gal-lupeol liposomes exhibited an average particle size of about 100 nm. The encapsulation efficiency was greater than 85%. The encapsulation efficiency of lupeol liposome and Gal-lupeol liposome, stored with 15% sucrose as glycoprotein for 6 months, was higher than 80%; although the particle size increased, they remained within 200 nm. The cell-uptake study demonstrated that the Gal-lupeol-liposome uptake efficiency was the highest in HepG2 cells. The HepG2 cells treated with the Gal-lupeol liposomes had higher apoptotic efficiency than the lupeol liposome and free lupeol. After HepG2 cells were treated with Gal-lupeol liposome, the expressions of AKT/mTOR-related proteins (p-AKT308 and p-AKT473) were also significantly reduced than the lupeol-liposome and free lupeol group. The in vivo targeting studies showed that Gal-NR-L exhibited liver-targeting effects on FVB mice. The pharmacodynamic study was performed by transfecting AKT and c-MET via the high-pressure tail vein of FVB mice. After Gal-lupeol-L administration, the liver index and liver weight of mice were less than those non-targeted group. The histopathological study showed that the lobular structure in the mice liver was clearer, the vacuoles were more obvious, and the cytoplasm was more abundant after Gal-lupeol-L administration. Also, the qRT-PCR study showed that AFP, GPC3, and EpCAM mRNA expression levels were significantly lower than those non-targeted lupeol-liposomes.

A homogenous polysaccharide from Codonopsis pilosula (CPP) was isolated and identified to be an inulin-type fructan, coded as CPP1-2-1. The polysaccharide exhibited anti-inflammatory effect against lipopolysaccharide (LPS) induced RAW264.7 cells in vitro and dextran sodium sulfate (DSS)-induced colitis mice in vivo. Moreover, the expression of cytokines was further detected by qPCR, and the results showed that the polysaccharides can reduce the expression of inflammatory factors such as TLR4, NF-κB, TNF-α and IL-6 in the cells, indicating the anti-inflammatory activities may be achieved by inhibiting the activation of TLR4/NF-κB pathway.

Our previous study proved that celastrol was a potential candidate for hepatocellular carcinoma (HCC) therapy. However, poor water solubility and toxic side effects may restrict its clinical application. To overcome these shortcomings and optimize its antitumor efficacy, we developed galactosylated liposomes using galactose-modified 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-poly(ethylene glycol) to deliver celastrol (C-GPL). C-GPL improved the water solubility of celastrol and exhibited high encapsulation efficiency, good stability in serum, and slow drug release profile. In vitro studies showed that C-GPL increased the cellular uptake of celastrol through receptor-mediated endocytosis, thereby enhancing celastrol cytotoxicity and cancer cell apoptosis. Particularly, in vivo antitumor activity of C-GPL was assessed in rapid HCC mouse models established via hydrodynamic transfection of the activated forms of AKT and c-Met. Compared to free celastrol, C-GPL significantly prevented liver weight gain, decreased liver damage biomarkers (glutamic-oxalacetic transaminase and alanine aminotransferase) and HCC marker (alpha-fetoprotein), and led to tumor disappearance on the liver surface. The improved therapeutic effect of C-GPL may be attributed to suppression of AKT activation, induction of apoptosis, and retardation of cell proliferation. Importantly, C-GPL exerted low toxicity to normal tissues without causing severe weight loss in mice. Taken together, C-GPL may become a promising drug delivery system for HCC treatment.

Background: Oxidative stress (OS) plays an important role in the progression of many aging-related diseases. Exercises can delay this kind of progress, but aerobic exercise is the most commonly used type of training among older adults; therefore, its influence needs to be further verified.Methods: A literature search was conducted in eight electronic databases, including Cochrane, EMBASE, PubMed, Web of Science, China National Knowledge Infrastructure (CNKI), China Science and Technology Journal Database (VIP), Wanfang Date, and SinoMed from their inception to April 2020. Methodological quality was assessed using Cochrane RoB tool v2.0 for individual studies, and RevMan 5.3 software was used to perform the meta-analysis.Results: The meta-analysis included 20 studies, involving 1,170 older adults. The results showed that regular aerobic exercise could reduce blood oxidant markers, including malondialdehyde (MDA; SMD=−1.80, 95% CI −2.46 to −1.14, pp=0.02), and increase the levels of antioxidant factors, such as nitric oxide (NO; SMD=0.89, 95% CI 0.37–1.41, pp=0.001), and total antioxidant capacity (TAC; SMD=1.22, 95% CI 0.45–1.98, p=0.002), with clear statistical significance. It may also improve the levels of other OS markers, such as 8-OHdG, 8-isoPGF2, VE, and reduced glutathione/oxidized glutathione (GSH/GSSG).Conclusion: Regular aerobic exercise may have a positive effect on the OS levels of older adults by reducing some oxidant markers and increasing antioxidant marker levels.

Objective: The objective of this study was to enhance the solubility and bioavailability of Lupeol.Methods: Utilizing a thin-film dispersion method, we prepared Lupeol-loaded PEGylated liposomes an...

Hepatocellular carcinoma (HCC) is a lethal malignancy with few effective options for therapeutic treatment in its advanced stages. Metformin, a first-line oral agent used in the treatment of type 2 diabetes, exhibits efficacy in metabolic reprogramming fueling changes in cell growth and proliferation for multiple cancer types, including HCC. However, the molecular mechanism by which metformin delays hepatocarcinogenesis in individuals with hepatic steatosis remains rare. Here, we investigate the preventive efficacy of metformin in a rapid AKT/c-Met-triggered HCC mouse model featuring excessive levels of steatosis. Hematoxylin and eosin staining, Oil Red O staining and immunoblotting were applied for mechanistic investigations. Pharmacological and biochemical strategies were employed to illuminate molecular evidence for HCC cell lines. The results show that metformin obstructs the malignant transformation of hepatocytes in AKT/c-Met mice. Mechanistically, metformin reduces the expression of phospho-ERK (Thr202/Tyr204) and two forms of proto-oncogenes, Cyclin D1 and c-Myc, in AKT/c-Met mice. Moreover, metformin ameliorates FASN-mediated aberrant lipogenesis and HK2/PKM2-driven ATP generation in vivo. Furthermore, metformin represses the expression of FASN and HK-2 by targeting c-Myc in an AMPK-dependent manner in vitro. In addition, metformin is effective at inhibiting PKM2 expression in the presence of an AMPK inhibitor compound C, suggesting that its functioning in PKM2 is AMPK-independent. Our study experimentally validates a novel molecular mechanism by which metformin alleviates enhanced lipogenesis and high energy metabolism during hepatocarcinogenesis, indicating that metformin may serve as an agent for the prevention of HCC in patients with nonalcoholic fatty liver diseases.

Size controllable silver nanoparticles (AgNPs) were synthesized in situ on the polysaccharides-based nanotubes, which were formed by the triple-helix polysaccharide extracted from black fungus (AF1). The results of transmission electron microscope (TEM), scanning electron microscope (SEM) and energy dispersive spectrometer (EDS) proved that AgNPs with the size from 10-25 nm were uniformly dispersed on the surface of AF1 dendritic nanotubes without affecting their tubular morphology. Moreover, due to the tubular structure, the loaded silver content of the composites (AgNPs and AF1 nanotube, AF1-Ag) could reach about 50% by thermogravimetric analysis (TG) evaluation. Thus, the smaller size of AgNPs and higher silver loading content suggest that the composites could be applied in the biomedical field. The antibacterial properties of AF1-Ag were evaluated as an example in the present work. As expected, the culture medium contained a few of AF1-Ag (10% ω%, c = 50 μg/mL) exhibited obvious antibacterial properties, and the effect of bacteriostasis increased with the increase of the amount of supported silver content. Taken together, the AF1-Ag with good antibacterial activity and good stability has the potential to be applied in the antibacterial field.

Urolithin A (Uro-A), a metabolite of ellagitannins in mammals’ intestinal tract, displays broad biological properties in preclinical models, including anti-oxidant, anti-inflammatory, and anti-tumor effects. However, the clinical application of Uro-A is restricted because of its low aqueous solubility and short elimination half-life. Our purpose was to develop a delivery system to improve the bioavailability and anti-tumor efficacy of Uro-A. To achieve this goal, urolithin A–loaded PEGylated liposomes (Uro-A-PEG-LPs) were prepared for the first time and its physicochemical properties and anti-tumor efficacy in vitro were evaluated. The morphology of Uro-A-PEG-LPs displayed a uniform sphere under transmission electron microscope. The particle size, polydispersity index, zeta potential, and encapsulation efficiency of Uro-A-PEG-LPs were 122.8 ± 7.4 nm, 0.25 ± 0.16, − 25.5 ± 2.3 mV, and 94.6 ± 1.6%, respectively. Moreover, Uro-A-PEG-LPs possessed higher stability and could be stably stored at 4°C for a long time. In vitro release characteristics indicated that Uro-A-PEG-LPs possessed superior sustained release properties. The results of confocal laser scanning microscopy experiment showed that the coumarin 6–loaded PEGylated liposomes (C6-PEG-LPs) have superior cellular uptake than that of conventional liposomes. In addition, in vitro tests demonstrated that Uro-A-PEG-LPs elevated cytotoxicity and pro-apoptotic effect in human hepatoma cells comparing with free Uro-A. Furthermore, the results of pharmacokinetic experiments showed that the t1/2, AUC0-t, and MRT0-t of Uro-A-PEG-LPs increased to 4.58-fold, 2.33-fold, and 2.43-fold than those of free Uro-A solution, respectively. Collectively, these manifested that PEGylated liposomes might be a potential delivery system for Uro-A to prolonging in vivo circulation time, promoting cellular uptake, and enhancing its anti-tumor efficacy.

Increasing the drug tumor-specific accumulation and controlling their release is considered one of the most effective ways to increase the efficacy of drugs. Here, we developed a vesicle system that can target hepatoma and release drugs rapidly within tumor cells. This non-ionic surfactant vesicle is biodegradable. Galactosylated stearate has been used to glycosylate the vesicles to achieve liver targeting; replacement of a portion (Chol:CHEMS = 1:1) of cholesterol by cholesteryl hemisuccinate (CHEMS) allows for a rapid release of drugs in an acidic environment. In vitro release experiments confirmed that galactose-modified pH-sensitive niosomes loaded with tanshinone IIA had excellent drug release performance in acid medium. In vitro experiments using ovarian cancer cells (A2780), colon cancer cells (HCT8), and hepatoma cell (Huh7, HepG2) confirmed that the preparation had specific targeting ability to hepatoma cells compared with free drugs, and this ability was dependent on the galactose content. Furthermore, the preparation also had a more substantial inhibitory effect on tumor cells, and subsequent apoptosis assays and cell cycle analyses further confirmed its enhanced anti-tumor effect. Results of pharmacokinetic experiments confirmed that the vesicle system could significantly extend the blood circulation time of tanshinone IIA, and the larger area under the curve indicated that the preparation had a better drug effect. Thus, the results of biodistribution experiments confirmed the in vivo liver targeting ability of this preparation. Niosomes designed in this manner are expected to be a safe and effective drug delivery system for liver cancer therapy.

Camellia oleifera Abel. is a critical oil tree species. Camellia oil, which is extracted from the seeds, is widely regarded as a premium cooking oil, with the content of oleic acid being over 80%. Light is thought to be one of the largest essential natural components in the regulation of plant developmental processes, and different light qualities can considerably influence plant physiological and phenotypic traits. In this research, we examined the growth and physiological responses of C. oleifera &ldquo, MIN 43&rdquo, cultivar plantlets to three different wavelengths of light, containing white, red, and blue light, and we utilized the combination of the PacBio single-molecule real-time (SMRT) and Illumina HiSeq RNA sequencing to obtain the mRNA expression profiles. The results showed that plantlets growing under blue light conditions displayed superior growth performance, including stimulated enhancement of the leaf area, increased leaf number, increased chlorophyll synthesis, and improved photosynthesis. Furthermore, SMAT sequencing created 429,955 reads of inserts, where 406,722 of them were full-length non-chimeric reads, and 131,357 non-redundant isoforms were produced. Abundant differentially expressed genes were found in leaves under different light qualities by RNA-sequencing. Gene expression profiles of actin, dynein, tubulin, defectively organized tributaries 3 (DOT3), and ADP ribosylation factor 5 (ARF5) were associated with the greatest leaf performance occurring under blue light conditions. Moreover, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis identified hundreds of pathways involved in different light conditions. The pathways of the plant circadian rhythm and hormone signal transduction were associated with different light quality responses in C. oleifera. Phytochrome B (PHYB), constitutively photomorphogenic 1 (COP1), long hypocotyl 5 (HY5), auxin/indole-3-acetic acid (AUX/IAA), Gretchen Hagen 3 (GH3), and small auxin-up RNA (SAUR), which were differentially expressed genes involved in these two pathways, play a vital role in responses to different wavelengths of light in C. oleifera. In addition, blue light significantly promotes flavonoid biosynthesis via changing expression of related genes.

Natural plants are considered as a huge treasure for anticancer. Amomum tsaoko, a plant of Zingiberaceae, is used widely as a food and traditional medicine in East Asia. In previous studies, Amomum tsaoko has antitumor effect on liver cancer cells, but the mechanism is not clear. Here, we demonstrated that ethanol extract from Amomum tsaoko (At-EE) could inhibit ovarian cancer and decrease angiogenesis in vivo. At-EE did not influence vascular endothelial cells directly, but decreased IL-6 and VEGF secreted by ovarian cancer cells to inhibit angiogenesis through inhibition of p-STAT3 and NF-kB activation. In addition, we demonstrated that p-STAT3 and NF-kB could adjust each other and IL-6 and VEGF also mediate p-STAT3 and NF-kB too, which created a loop. In addition, At-EE interrupted p-STAT3/NF-kB/IL-6 and VEGF loop through induced ER stress. These results reveal that p-STAT3/NF-kB/IL-6 and VEGF is a cascade amplification loop in ovarian cancer for angiogenesis, and induced ER stress can interrupt it. Taken together, this work explored the anticancer activities of Amomum tsaoko, which could be a potential therapeutic candidate in the treatment of ovarian cancer.

Although the suppressing effects of celastrol on hepatocellular carcinoma (HCC) have been demonstrated, evidence for the targeting of fatty acid synthetase (FASN) in the development of HCC by celastrol is still rare. In this study, the effect of celastrol on a rapid HCC model featuring co-activation of AKT/c-Met oncogenes in mice was studied. The effect of celastrol on the alpha-fetoprotein level in the liver and serum was also investigated. Protein expressions of PCNA, Ki67 and FASN in celastrol-treated AKT/c-Met HCC mice were observed. The molecular mechanism of celastrol on the AKT/c-Met signaling pathway was elucidated. The results revealed that celastrol significantly repressed the AKT/c-Met induced HCC development and down-regulated the mRNA expression of AFP in the liver and the AFP level in serum. Furthermore, the expression of proliferation-associated proteins in the HCC tissues was reduced by celastrol treatment. Moreover, the abundant steatosis that resulted from FASN accumulation in the liver in AKT/c-Met mice was also attenuated. Finally, the phosphorylation of AKT and ERK1/2 in HCC mice was repressed by celastrol treatment. Overall, our data demonstrate that celastrol exerts an antiproliferative and novel lipid-decreasing effect by targeting AKT/ERK and FASN in HCC development in vivo.

In this paper, the peptide amphiphiles (PA) which consists of RGDSEEEEEEEEEEK as pH-sensitive segment and stearic acid as hydrophobic segment named RGDS-E10-Lys(C18) was successfully synthesized. TEM images showed that uniformly dispersed nanoparticles could be formed by PA molecules in pH 7.4 medium, however, disintegrated in pH 5.0 medium. Circular dichroism (CD) spectrum indicated that polypeptide adopted a random-coil conformation in neutral medium (pH 7.4). The CD signal was significantly attenuate for decreased solubility of PA in medium with pH 5.0. As expected, the prepared RGDS-E10-Lys(C18) assembly showed high pH-sensitive property which demonstrated a much more rapid drug release from micelles in tumor tissue (acidic environment) than in physiological environment (neutral environment). After DOX-loaded micelles incubated with tumor cells, the cytotoxicity of the micelles against Hela cells was increased obviously, indicating the great potential of micelles developed here as promising ve...

Polymeric delivery vehicles can improve the safety and efficacy of chemotherapy drugs by facilitating preferential tumor delivery. Double hydrophilic block copolymer (DHBC)-based prodrugs are considered as ideal candidates for drug delivery due to the elegant integration of benefits from both structures including polymeric prodrugs’ superior protection and minimal premature drug release using covalent links and a DHBC-based “green” self-assembly strategy by a simple stimulus in a pure aqueous phase without the use of any organic solvent. Clearly, the location of drug molecules in the polymeric prodrugs has exerted a significant effect on their therapeutic efficiency. However, there has been no published data so far, to our knowledge, reporting the effect of drug-conjugated sites on its therapeutic efficacy, as well as some basic guidelines that can be followed to direct the future design of polymeric prodrugs. To this end, herein a thermo-sensitive DHBC, poly(N-(2-hydroxypropyl) methacrylamide)-b-poly(N-isopropyl acrylamide) (P(HPMA)-b-P(NIPAAm)), was designed and synthesized by successive reversible addition and fragmentation chain transfer (RAFT) polymerizations, and was chosen as a platform to clarify this issue. An anti-cancer drug, doxorubicin (DOX) was conjugated to the hydrophilic PHPMA block and the temperature-responsive P(NIPAAm) block, respectively, through a pH-liable hydrazone bond to fabricate two different types of polymeric prodrugs with the drug tethered to the micellar hydrophilic PHPMA shell or encapsulated within the hydrophobic P(NIPAAm) core upon temperature elevation above its lower critical solution temperature (LCST). A detailed comparison study was carried out to investigate which structure exhibits better properties and higher therapeutic efficacy in terms of micellar size, stability, cellular uptake, drug loading capacity, drug release behaviors and cell viability. The results showed the self-assembly of both DHBC-based prodrugs into well-dispersed spherical micelles with similar average hydrodynamic diameters (Dh) around 150 nm in phosphate buffer (PBS, pH 7.4) at 37 °C, but a higher drug loading content (DLC), and enhanced pH-mediated drug release, i.e., much accelerated drug release at pH 5.0, while slower at pH 7.4, as well as enhanced cytotoxicity when the drug was conjugated to the hydrophilic shell of the micelles. The guidelines obtained in this study are thus believed to direct the future design and development of polymeric prodrugs for efficient cancer therapy.

A nanocarrier system of methoxypolyethylene glycol amine (mPEG-NH2) functionalized polydopamine (PDA) coated hollow mesoporous silica nanoparticles (HMSNs-PDA-PEG) was developed with pH-responsive, which combined doxorubicin hydrochloride (DOX) and quercetin (QUR) to reverse multidrug resistance (MDR) and improved anticancer effects on taxol (TAX) and DOX double resistant human colorectal cancer cell line HCT-8 (HCT-8/TAX cells). Well-dispersed nanoparticles (HMSNs-PDA-PEG) were prepared with a dimension of around 170 nm. The surface morphology and chemical properties of HMSNs-PDA-PEG were also successfully characterized by transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), thermal gravimetric analysis (TGA), X-ray diffraction (XRD), Brunauer-Emmett-Teller (BET) method, Fourier transform infrared spectroscopy (FT-IR) and dynamic light scattering (DLS). Drug release experiments results indicated that DOX and QUR (QD) loaded nanoparticles (HMSNs-PDA-PEG@QD) had similar release kinetic profiles of each drug, which all exhibited highly sensitive to pH value due to the surface PDA coating. Additionally, the HCT-8 cells or HCT-8/TAX cells were employed to assess the cellular uptake and cytotoxicity of various drug-free or drug-loaded HMSNs samples. Meanwhile, a series of biological evaluations demonstrated that the HMSNs-PDA-PEG@QD exhibited remarkable ability to overcome MDR compared with free DOX and HMSNs-PDA-PEG@DOX. Taken together, these results revealed that HMSNs-PDA-PEG@QD was suitable as a prospective and efficient drug delivery nanosystem for overcoming multidrug resistance.

Arsenic trioxide (As2O3, ATO), a FDA approved drug for hematologic malignancies, was proved of efficient growth inhibition of cancer cell in vitro or solid tumor in vivo. However, its effect on solid tumor in vivo was hampered by its poor pharmacokinetics and dose-limited toxicity. In this study, a polyacrylic acid capped pH-triggered mesoporous silica nanoparticles was conducted to improve the pharmacokinetics and enhance the antitumor effect of arsenic trioxide. The mesoporous silica nanoparticles loaded with arsenic trioxide was grafted with polyacrylic acid (PAA-ATO-MSN) as a pH-responsive biomaterial on the surface to achieve the release of drug in acidic microenvironment of tumor, instead of burst release action in circulation. The nanoparticles were characterized with uniform grain size (particle sizes of 158.6 ± 1.3 nm and pore sizes of 3.71 nm, respectively), historically comparable drug loading efficiency (11.42 ± 1.75%), pH-responsive and strengthened sustained release features. The cell toxicity of amino groups modified mesoporous silica nanoparticles (NH2-MSN) was significantly reduced by capping of polyacrylic acid. In pharmacokinetic studies, the half time (t1/2β) was prolonged by 1.3 times, and the area under curve) was increased by 2.6 times in PAA-ATO-MSN group compared with free arsenic trioxide group. Subsequently, the antitumor efficacy in vitro (SMMC-7721 cell line) and in vivo (H22 xenografts) was remarkably enhanced indicated that PAA-ATO-MSN improved the antitumor effect of the drug. These results suggest that the polyacrylic acid capped mesoporous silica nanoparticles (PAA-MSN) will be a promising nanocarrier for improving pharmacokinetic features and enhancing the anti-tumor efficacy of arsenic trioxide.

Orlistat (Xenical™), a US Food and Drug Administration (FDA)-approved anti-obesity drug, shows efficacy against multiple tumor types, including hepatocellular carcinoma (HCC), due to its ability to inhibit fatty acid synthase (FASN) activity. However, whether orlistat affects hepatocellular malignant transformation during hepatocarcinogenesis in vivo is unknown. This study assessed the antisteatotic and antitumorigenic efficacy of orlistat in a rapid HCC FVB/N mouse model established via hydrodynamic transfection of activated forms of AKT and c-Met proto-oncogenes. Human hepatoma cell lines were used for mechanical validation in vitro. Hematoxylin and eosin staining, immunohistochemistry, and immunoblotting were applied for the mechanistic investigation. The results revealed that when orlistat was administered in the early stage of AKT/c-Met-triggered hepatocarcinogenesis, it resulted in the elimination of hepatic tumor burden. Mechanistically, orlistat efficiently elevated PTEN expression and suppressed AKT/SREBP1/FASN signaling both in vivo and in vitro, impairing AKT/c-Met-driven de novo lipogenesis and aberrant proliferation. Altogether, this study demonstrates the antilipogenic and antiproliferative efficacy of orlistat in hepatocarcinogenesis, suggesting that orlistat may be beneficial for the treatment of HCC, especially in NAFLD-related HCCs featuring activated AKT/mTOR cascade and increased lipogenesis in livers.

Hepatocellular carcinoma (HCC) is one of the most common fatal malignancies worldwide. Inhibition of the lipogenic enzymes involved in hepatic de novo lipogenesis can both effectively restrain proliferation of HCC cells in vitro and reduce the risk of hepatocarcinogenesis in vivo. Although a natural coumarin derivative osthole shows efficacy in suppressing cell proliferation and inducing apoptosis in cultured hepatoma cells and HCC xenograft tumors, the molecular mechanism by which osthole delays hepatocellular malignant transformation during lipogenesis-driven hepatocarcinogenesis remains unknown. Here, we evaluate the efficacy of osthole in a rapid HCC mouse model featuring excessive levels of hepatic steatosis established via hydrodynamic transfection of activated forms of AKT and c-Met proto-oncogenes. Moreover, human hepatoma cell lines were employed for in vitro assessment. Hematoxylin and eosin staining, immunoblotting and immunohistochemistry were applied for mechanistic investigations. The results revealed that if osthole was administered in the early stage of AKT/c-Met-driven HCC, it led to disease stabilization. Moreover, osthole alleviated hepatic steatosis in the AKT/c-Met mice. Further evidence at the molecular level suggested that osthole reduced the expression of phosphor-extracellular signal-regulated kinase 1/2 (ERK1/2), proliferating cell nuclear antigen (PCNA) and Ki67 in livers of the AKT/c-Met mice. Mechanically, osthole efficiently repressed the phospho-AKT (Thr308) / ribosomal protein S6 (RPS6) / fatty acid synthase (FASN) signaling both in mice and in vitro. Altogether, this study suggests that osthole exerts its antilipogenic and antiproliferative efficacy by suppressing the AKT/FASN axis and ERK phosphorylation, which contributes to its capacity to delay hepatocarcinogenesis.

Background & Aims The progression of hepatosteatosis to non-alcoholic steatohepatitis (NASH) is a critical step in the pathogenesis of hepatocellular cancer. However, the underlying mechanism(s) for this progression is essentially unknown. This study was designed to determine the role of miR-378 in regulating NASH progression. Methods We used immunohistochemistry, luciferase assays and immunoblotting to study the role of miR-378 in modulating an inflammatory pathway. Wild-type mice kept on a high-fat diet (HFD) were injected with miR-378 inhibitors or a mini-circle expression system containing miR-378, to study loss and gain-of functions of miR-378. Results MiR-378 expression is increased in livers of dietary obese mice and patients with NASH. Further studies revealed that miR-378 directly targeted Prkag2 that encodes AMP-activated protein kinase γ 2 (AMPKγ2). AMPK signaling negatively regulates the NF-κB-TNFα inflammatory axis by increasing deacetylase activity of sirtuin 1. By targeting Prkag2, miR-378 reduced sirtuin 1 activity and facilitated an inflammatory pathway involving NF-κB-TNFα. In contrast, miR-378 knockdown induced expression of Prkag2, increased sirtuin 1 activity and blocked the NF-κB-TNFα axis. Additionally, knockdown of increased Prkag2 offset the inhibitory effects of miR-378 inhibitor on the NF-κB-TNFα axis, suggesting that AMPK signaling mediates the role of miR-378 in facilitating this inflammatory pathway. Liver-specific expression of miR-378 triggered the development of NASH and fibrosis by activating TNFα signaling. Ablation of TNFα in miR-378-treated mice impaired the ability of miR-378 to facilitate hepatic inflammation and fibrosis, suggesting that TNFα signaling is required for miR-378 to promote NASH. Conclusion MiR-378 plays a key role in the development of hepatic inflammation and fibrosis by positively regulating the NF-κB-TNFα axis. MiR-378 is a potential therapeutic target for the treatment of NASH. Lay summary The recent epidemic of obesity has been associated with a sharp rise in the incidence of non-alcoholic fatty liver disease (NAFLD). However, the underlying mechanism(s) remains poorly described and effective therapeutic approaches against NAFLD are lacking. The results establish that microRNA-378 facilitates the development of hepatic inflammation and fibrosis and suggests the therapeutic potential of microRNA-378 inhibitor for the treatment of NAFLD.

Various benefits of flavonoids for ameliorating cardiovascular diseases have been demonstrated. However, the lowering effects on blood pressure caused by antiproliferative potentials of flavonoids in vascular smooth muscle cells are rare. In this study, the antihypertensive effects of total flavonoids from Ampelopsis megalophylla were investigated. The dynamic pressure values and the rate of media thickness versus lumen diameter were measured by the tail-cuff system and H&E staining in vivo, respectively. The mRNA expressions of ACE, Ang II, eNOS, c-Myc, cyclin D1 and p27Kip1 in thoracic aorta or A7r5 cells were measured by qPCR, respectively. The protein expressions of c-Myc, Cyclin D1, p27Kip1 and β-catenin in tissues or A7r5 cells were measured by Western blot assay. Total flavonoids of A. megalophylla (TFAM) reduced the expressions of ACE and Ang II, and elevated the content of eNOS in thoracic aorta cells of SHRs. Furthermore, TFAM decreased the mRNA and protein expressions of c-Myc and cyclin D1 by repressing the Wnt/β-catenin-mediated TCF/LEF transcriptional activation both in vivo and in vitro, which is synergetic with the up-regulation of p27Kip1 expression. Our study provided evidence for developing flavonoids from A. megalophylla as herbal supplements to prevent against cardiovascular diseases by suppressing vascular remodeling.

An abnormality in the Lin28/let-7a axis is relevant to the progression of hepatitis B virus (HBV)-positive hepatocellular carcinoma (HCC), which could be a novel therapeutic target for this malignant tumor. The present study aimed to investigate the antiproliferative and anti-invasive effects of urolithin A in a stable full-length HBV gene integrated cell line HepG2.2.15 using CCK-8 and transwell assays. The RNA and protein expressions of targets were assessed by quantitative PCR and western blot, respectively. Results revealed that urolithin A induced cytotoxicity in HepG2.2.15 cells, which was accompanied by the cleavage of caspase-3 protein and down-regulation of Bcl-2/Bax ratio. Moreover, urolithin A suppressed the protein expressions of Sp-1, Lin28a, and Zcchc11, and elevated the expression of microRNA let-7a. Importantly, urolithin A also regulated the Lin28a/let-7a axis in transient HBx-transfected HCC HepG2 cells. Furthermore, urolithin A decelerated the HepG2.2.15 cell invasion, which was involved in suppressing the let-7a downstream factors HMGA2 and K-ras. These findings indicated that urolithin A exerted the antiproliferative effect by regulating the Lin28a/let-7a axis and may be a potential supplement for HBV-infected HCC therapy.

Calcium has been associated with improved cold tolerance in many crops. The aim of this study was to investigate the changes in leaf cell Ca 2+ distribution and cell organelle ultrastructure of loquat (Eriobotrya japonica Lindl.) plants in response to cold stress at -3℃, using transmission electron microscopy (TEM). Two loquat accessions, Zaozhong 6 (a commercial cultivar) and oakleaf loquat (a wild relative) were used. Cold tolerance, as measured by leaf browning rate, was higher in oakleaf plants, and calcium treatment improved cold tolerance in both species. Cold stress first induced inward transport of Ca 2+ from the intracellular space. Then, the imported Ca 2+ was aggregated around the chloroplast membrane, finally entering the chloroplast. This pattern of Ca 2+ distribution in leaf cells occurred earlier in Zaozhong 6 than in the wild loquat. With increasing time of cold exposure, the chloroplast membranes of Zaozhong 6 leaves were damaged, blurred and even disappeared, while those of wild oakleaf loquat leaves maintained their structure longer. In Zaozhong 6, cold stress induced a clear cavity between poorly structured granal thylakoids and vesicles appearing inside the chloroplast, while in oakleaf leaves cold stress had little effect on the ultrastructure of chloroplasts (although chloroplast membranes looked blurred). Loquat leaves accumulated free calcium ions around chloroplasts in response to cold stress, with earlier calcium accumulation occurring in the cold-sensitive cultivar Zaozhong 6 than in wild oakleaf loquat. These results demonstrate that these two loquat species have differences in both cold tolerance and calcium accumulation dynamics.

The circulating level of platelet-activating factor acetylhydrolase (PAF-AH) is a novel biomarker to predict the presence of coronary heart disease. PAF-AH gene polymorphisms may be responsible for the variance of circulating PAF-AH levels in individuals. However, the association of PAF-AH gene polymorphisms with circulating PAF-AH levels and the susceptibility to coronary heart disease (CHD) remains unsolved. Blood stasis syndrome (BSS) of CHD is the most common type of TCM syndromes, and a previous study discovered its relationship with the elevated circulating PAF-AH levels. However, the association of gene polymorphisms and CHD with BSS is unclear at present. In this study, four polymorphisms (R92H, I198T, A379V, V279F) of the PAF-AH gene were genotyped in 570 CHD patients, of which 299 had BSS. In addition, 317 unaffected individuals from the same hospitals served as controls. Plasma PAF-AH levels were measured in 155 controls and 271 CHD patients selected randomly, including 139 CHD patients with BSS. In the Chinese Han population, plasma PAF-AH levels in CHD patients with BSS or without BSS were significantly higher (12.9 ± 6.5 and 11.1 ± 5.0 μM, respectively) than in controls (9.3 ± 5.2 μM); this difference still remained significant after adjustment for traditional risk factors or the inflammatory factors. The R92H polymorphism was highly related to the plasma PAF-AH levels and the risk of CHD, especially among patients with BSS, even with the adjustment for the effects of traditional factors. The I198T polymorphism was highly associated with risk of CHD with BSS, but was associated with neither the risk of CHD with no BSS nor with elevated plasma PAF-AH levels.

Activation of the AKT/mTOR cascade and overexpression of c-Met have been implicated in the development of human hepatocellular carcinoma (HCC). To elucidate the functional crosstalk between the two pathways, we generated a model characterized by the combined expression of activated AKT and c-Met in the mouse liver. Co-expression of AKT and c-Met triggered rapid liver tumor development and mice required to be euthanized within 8 weeks after hydrodynamic injection. At the molecular level, liver tumors induced by AKT/c-Met display activation of AKT/mTOR and Ras/MAPK cascades as well as increased lipogenesis and glycolysis. Since a remarkable lipogenic phenotype characterizes liver lesions from AKT/c-Met mice, we determined the requirement of lipogenesis in AKT/c-Met driven hepatocarcinogenesis using conditional Fatty Acid Synthase (FASN) knockout mice. Of note, hepatocarcinogenesis induced by AKT/c-Met was fully inhibited by FASN ablation. In human HCC samples, coordinated expression of FASN, activated AKT and c-Met proteins was detected in a subgroup of biologically aggressive tumors. Altogether, our study demonstrates that co-activation of AKT and c-Met induces HCC development that depends on the mTORC1/FASN pathway. Suppression of mTORC1 and/or FASN might be highly detrimental for the growth of human HCC subsets characterized by concomitant induction of the AKT and c-Met cascades.

To develop an effective method to remove and mineralize trace organic pollutants, a series of composite functional membranes containing immobilized ACF (activated carbon fibre) powder ( 3+ -TiO 2 , a combination of adsorbent and photo-catalyst on terylene (PET) filter cloth, was prepared by using a sol–gel and dip coating method. As high as 93% BPA (bisphenol A) pollutant can be separated successfully by adsorption onto the membrane in a series of batch adsorption tests, 40% removal was achieved in dynamic filtration/adsorption test, which also removed 98% koalinite suspended solids at the same time. The measured adsorption of BPA corresponds to 37.0, 70.6 and 62.2 mg (BPA)/g(ACF) for three membranes A, B and C, where the weight ratio of ACF to ACF + TiO 2 was 29.9%, 17.6% and 11.3% respectively in batch adsorption tests (50 ml solution) and 25.48 mg (BPA)/g(ACF) for membrane A after 2 h adsorption in 200 ml solution. After dynamic filtration, the total adsorption capacity was 81.0 mg (BPA)/g(ACF) for membrane A. It is noteworthy that membrane adsorption can be regenerated by UV/Fenton treatment. Order of BPA adsorption quantity is 1st > 2nd > 3rd time use for membrane C, due to loss of ACF resulted from the supersonic treatment before oxidative regeneration. Introducing TEOS in the preparation of sol–gel solution and subsequent coating to form composite membrane, improved membrane adsorption stability in UV/Fenton regeneration treatment.

Two new steroidal glycosides named riparosides A (1) and B (2), and two aromatic compounds (3, 4), together with four known flavonoid derivatives have been isolated from the EtOH extract of the rhizomes and roots of Smilax riparia A. DC. The structure of riparoside A (1) was determined to be 3-O-alpha-L-rhamnopyranosyl-(1--2)-[alpha-L-rhamnopyranosyl-(1--6)]-beta-D-glucopyranosyl 3beta,20alpha-dihydroxy-5alpha-furost-22(23)-ene 26-O-beta-D-glucopyranoside. Riparoside B (2) was characterized as 3-O-alpha-L-rhamnopyranosyl-(1--2)-[alpha-L-rhamnopyranosyl-(1--6)]-beta-D-glucopyranosyl 3beta,16beta-dihydroxy-5alpha-pregnan-20-one 16-O-[5-O-beta-D-glucopyranosyl 5-hydroxy-4-methyl-pentanoic acid]-ester 26-O-beta-D-glucopyranoside. Compounds 3 and 4 were elucidated as a sucrosyl ferulic acid ester and 7-O-methyl-10-oxythymol gentiobioside, respectively.

Thermoresponsive amphiphilic copolymer, poly[N-isopropyl acrylamide-co-3-(trimethoxysilyl)propylmethacrylate]-b-poly{N-[3-(dimethylamino)propyl]methacrylamide} with a branched structure was designed and synthesized by consecutive reversible addition–fragmentation chain-transfer polymerization. The further hydrolysis of trimethoxysilyl functions in 3-(trimethoxysilyl) propyl methacrylate units led to the fabrication of core-crosslinked (CCL) micelles with silica crosslinks at temperatures above the lower critical solution temperature of the poly(N-isopropyl acrylamide) block. The thermally induced structural and morphological changes of the CCL micelles in aqueous solution were investigated by transmission electron microscopy and 1H-NMR analyses. The resulting CCL micelles were further explored as nanocarriers for the codelivery of an anticancer drug and nucleic acid for enhanced therapeutic efficacy. The CCL micelles effectively condensed the nucleic acid and mediated higher gene transfer in the presence of serum than in serum-free transduction. A cytotoxicity study revealed that whereas the pure CCL micelles exhibited unapparent cytotoxicity, the codelivery of p53 and doxorubicin with the CCL micelle formulation resulted in better treatment efficiency than sole chemotherapy. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 41752.

The purpose of the study was to explore the association between plasma platelet activating factor (PAF) and platelet activating factor acetylhydrolase (PAF-AH) levels and risk of coronary heart disease (CHD) or blood stasis syndrome (BSS) of CHD. Questionnaire, routine clinical assays and plasma levels of PAF, PAF-AH and inflammatory factors hs-CRP and IL-6 were investigated or measured for 120 controls and 150 CHD patients (66 non-BSS and 84 BSS). Plasma PAF levels were higher in CHD patients [49.7 (34.8–73.2 pg/mL)] than in controls [23.8 (14.9–42.3 pg/mL)] (P < 0.001), and in BSS [56.0 (40.1–86.1 pg/mL)] than in non-BSS [47.4 (29.0–67.4 pg/mL)] (P = 0.027). Similarly, plasma PAF-AH levels were higher in CHD patients [11.5 (7.5–15.6 μmol/L)] than in controls [8.1 (5.4–12.6 μmol/L)] (P < 0.001), and in BSS [13.4 (8.7–18.5 μmol/L)] than in non-BSS [9.5 (7.3–14.3 μmol/L)] (P = 0.014). After adjustment for the confounded effects of inflammatory factors or conventional risk factors, plasma PAF and PAF-AH levels still had a significant difference between CHD patients and controls, but plasma PAF-AH rather than PAF levels had a significant difference between BSS and non-BSS. Elevated plasma PAF level contributed to the risk of CHD rather than BSS, and elevated plasma PAF-AH level was an independent risk factor of CHD and BSS.