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Background To improve responses to tumor microenvironments for achieving a better therapeutic outcome in combination cancer therapy, poly(e-caprolactone)-SS-poly(methacrylic acid) diblock copolymer (PCL-SS-PMAA) with a disulfide linkage between the hydrophobic and hydrophilic junctions was synthesized. Materials and methods Repeating units of PCL and PMAA in PCL-SS-PMAA were controlled and formulated into polymersomes (PSPps). Truncated octahedral Fe3O4 nanoparticles (IONPs) were synthesized and encapsulated to produce IONPs-PSPps NPs and doxorubicin (DOX) was further loaded to produce IONPs-PSPps@DOX NPs for theranostic applications. Results IONPs-PSPps NPs remained a superparamagnetic property with a saturation magnetization value of 85 emu⋅gFe3O4 -1 and a relaxivity value of 180 mM-1⋅s-1. Upon exposure to an alternating magnetic field (AMF), IONPs-PSPps NPs increased temperature from 25°C to 54°C within 15 min. Among test groups, the cell apoptosis was greatest in the group exposed to IONPs-PSPps@DOX NPs with AMF and magnet assistance. In vivo T2-weighted magnetic resonance images of A549 tumor-bearing mice also showed highest contrast and greatest tumor suppression in the tumor with AMF and magnet assistance. Conclusion IONPs-PSPps@DOX NPs are a potential theranostic agent having multifaceted applications involving magnetic targeting, MRI diagnosis, hyperthermia and chemotherapy.

A novel dual-stimuli cleavable linker containing adjacent UV light-sensitive o-nitrobenzyl ester and GSH-responsive disulfide bonds was first designed and synthesized to increase the responsivity to external stimuli. The functionalized linker was then utilized to prepare a dual-responsive amphiphilic block copolymer using ring-opening polymerization and atom transfer radical polymerization. The copolymer formed a micelle in an aqueous solution and showed dual-stimuli responses including photo-mediated cleavage under UV light irradiation at 365 nm as well as reduction-responsive degradation in the presence of a reducing agent. The micelle was nontoxic against three cell lines and majorly internalized via clathrin-mediated endocytosis. Doxorubicin (Dox) was loaded in the hydrophobic core of the micelle. Enhancement of a cell-killing effect against A549 cells was clearly observed in the Dox-encapsulated micelle when exposed to UV light.

Epicutaneous (EC) sensitization with protein allergens is the most important sensitization route for atopic dermatitis. Plasmacytoid dendritic cells (pDCs) are characterized by massive secretion of interferon-α (IFNα). B6 mice are T helper type 1 (Th1)-prone and are representative of non-atopic humans, whereas BALB/c mice are Th2-prone and are representative of atopic humans. Here, we show that naive BALB/c mice contain a greater number of nonactivated pDCs in peripheral lymph nodes (LNs) than do naive B6 mice. Naive BALB/c mice also have more of the CD8α- subset in LNs than naive B6 mice. Moreover, in vivo depletion of pDCs during EC sensitization results in enhanced Th2 responses in BALB/c mice, but not in B6 mice. Mechanistically, when BALB/c mice undergo EC sensitization, there is an increase in pDCs entering draining LNs. These cells exhibit modest activation including comparable costimulation expression but increased cytokine expression compared with those of naive mice. In vivo depletion of pDCs during EC sensitization significantly increases the activation of dermal dendritic cells (dDCs) suggesting a regulatory effect on these cells. To this end, a suppressive effect of pDCs on conventional dendritic cells was also demonstrated in vitro. Further, in vivo blockade of IFNα by an anti-IFNAR antibody (Ab) or in vivo reduction of IFNα production of pDCs by anti-siglec-H Ab both resulted in enhanced activation of dDCs. Collectively, our results demonstrate that pDCs suppress Th2 responses induced by EC sensitization via IFNα-mediated regulation of dDCs.

Influence of hydrogen concentration and defects introduced by neutron irradiation on hydrogen diffusion in tungsten has been investigated by molecular dynamics simulation at elevated temperatures. Hydrogen diffusion is shown to be significantly restrained at high concentrations due to spontaneous formation of platelet-like hydrogen clusters. For neutron irradiation defects, self-interstitials, mono-vacancies and vacancy clusters are considered. By clustering and acting as dislocation loops, self-interstitials show considerable trapping effects on hydrogen, leading to the suppression of hydrogen effective diffusion and the change of diffusion model in which hydrogen mainly diffuses along dislocation lines instead of hopping between tetrahedral interstitial sites. Moreover, an equation connecting hydrogen diffusion parameters and the total length of dislocation loops is empirically established. Different influences of mono-vacancies and vacancy clusters on hydrogen diffusion have been carefully identified. With the same vacancy concentration, hydrogen diffusivity is lower with mono-vacancies than that with vacancy clusters because more isolated trapping sites are provided by mono-vacancies. This work is not only helpful for understanding the synergistic effects of neutron irradiation and plasma interaction, but also potentially applicable for larger scale simulations as input data.

Combining upconversion nanoparticles (UCNPs) and UV-sensitive polymers to form a smart drug delivery system (DDS) is a promising strategy to circumvent drawbacks of direct UV excitation in clinical applications. This study tuned up core-shell UCNPs with a shell thickness of 6 nm and emission wavelength falling in the ultraviolet region at 350 nm under near-infrared (NIR) light irradiation at 980 nm. An amphiphilic block copolymer with UV-responsive

The nutritive value of starch, the major source of dietary energy in pigs, varies depending on its susceptibility for digestion. The botanical origin of starch determines starch structure, and therefore, digestibility. To compare digestibility of starch, fiber, gross energy (GE), crude protein, and amino acid (AA), and to characterize undigested starch of grains in growing pigs, seven ileal-cannulated barrows (initial body weight, 30 kg) were fed six diets containing 96% of one of six test ingredients (three pulse grains: zero-tannin faba bean, green field pea, or mixed-cultivar chickpea; three cereal grains: hulled barley, hard red spring wheat, or hybrid yellow, dent corn), or a N-free diet in a 7 × 7 Latin square at 2.8 × maintenance digestible energy. Grain samples were ground with a hammer mill through a 2.78-mm screen. Amylose content ranged from 29% to 34% for pulse grains and from 22% to 25% for cereal grains. The apparent ileal digestibility (AID) of starch was greater (P < 0.05) in cereal (94% to 97%) than pulse grains (85% to 90%) and was lowest (P < 0.05) in faba bean (85.3%) followed by field pea (87.2%) and chickpea (90.1%). However, apparent total tract digestibility (ATTD) of starch of all tested grains was close to 100%. Apparent hindgut fermentability (AHF, as ATTD − AID) of starch was greater (P < 0.05) in pulse grains (9.9% to 15%) than cereal grains (3.3% to 4.8%). The AHF of total dietary fiber tended to be the greatest (P < 0.10) for corn (43.5%) and lowest for wheat (25.3%). The AHF of GE was greater (P < 0.05) in pulse grains (17% to 20%) than in cereal grains (9% to 11%) and resulted in greater (P < 0.05) digestible energy (DE) contribution from hindgut fermentation for pulse grains than cereal grains (0.9 vs. 0.5 Mcal/kg). Wheat had the greatest standardized ileal digestibility of total AA (90.2%; P < 0.05). Confocal laser scanning microscopy images revealed that 20% to 30% of starch granules of pulse grains were entrapped in protein matrixes. In scanning electron microscopy images, starch granules were larger in faba bean and field pea than cereal grains. Digesta samples revealed pin holes and surface cracks in starch granules of corn and wheat, respectively. In conclusion, hindgut fermentation of starch and fiber was greater in pulse grains than cereal grains resulting in a greater DE value despite lower ileal DE for pulse grain than cereal grains. Defining the digestible and fermentable fractions of starch may enhance the accuracy of equations to predict the net energy value of these feedstuffs.

There are certain disadvantages in treating bacterial infections through conventional methods. For this reason, the current study does focus on combating bacterial wound infections by photothermal therapy assisted by gold nanorod-peptide conjugates (GNR-peptide conjugates). Two peptides, the cationic antimicrobial peptide LL-37 and neuropeptide ANGIOPEP-2 both with specificity for targeted bacterial binding, were conjugated with GNR surface through electrostatic interactions. The GNR-peptide conjugates showed good biocompatibility, sufficient stability, enhanced targeting, potential photothermal killing of bacteria, and possible acceleration of wound healing. The photo-biomodulation properties of NIR improved the wound closure rates through enhanced cell migration. The multifunctional LL37-conjugated GNRs significantly enhanced photothermal therapeutic outcomes based on bacterial targeting with promising wound healing properties.

Paclitaxel (PTX) is a widely used chemotherapy drug; however, frequent use causes multidrug resistance (MDR), which limits the utility of PTX against advanced non-small-cell lung cancer (NSCLC). PTX-resistant subline (NCI-H23-TXR) was established in vitro by exposing NCI-H23 cells to gradually increased concentrations of PTX in culture medium. Distinct Beclin expression of autophagy level was observed between resistant NCI-H23-TXR and parental NCI-H23 cells. Beclin-small interfering RNA (siRNA) was selected to restore sensitivity of PTX against NCI-H23-TXR. Chondroitin sulfate-polyethylenimine (CS-PEI) was constructed for delivery and protection of Beclin-siRNA. To delineate the underlying molecular mechanism of Beclin knockdown, we analyzed different MDR expression proteins of two cells using western blot, and the corresponding genes were confirmed by real-time PCR. Compared with NCI-H23, NCI-H23-TXR had higher expression levels in P-glycoprotein (P-gp) and multidrug resistance protein 7 (ABCC10). Knockdown of Beclin simultaneously inhibited P-gp and ABCC10, and renewed the sensitivity of PTX against NCI-H23-TXR. Research on zebrafish embryos revealed that tumor sizes decreased in NCI-H23 tumor xenografts but remained intact in NCI-H23-TXR tumor xenografts as zebrafish were treated with 1 μg/mL PTX. In contrast, the tumor sizes decreased in NCI-H23-TXR tumor xenografts with zebrafish pre-transfected with CS-PEI/Beclin-siRNA followed by the same treatment of PTX. The role of autophagy was associated with MDR development. This study paves the way for a new avenue of PTX in MDR-related lung cancer therapy using CS-PEI as a gene delivery carrier. Keywords: autophagy, Beclin-siRNA, multidrug resistance, MDR, non-viral gene delivery vector, paclitaxel, PTX

Pseudomonas fluorescens is a typical spoiler of proteinaceous foods, and it is characterized by high spoilage activity. The sigma factor RpoN is a well-known regulator controlling nitrogen assimilation and virulence in many pathogens. However, its exact role in regulating the spoilage caused by P. fluorescens is unknown. Here, an in-frame deletion mutation of rpoN was constructed to investigate its global regulatory function through phenotypic and RNA-seq analysis. The results of phenotypic assays showed that the rpoN mutant was deficient in swimming motility, biofilm formation, and resistance to heat and nine antibiotics, while the mutant increased the resistance to H2O2. Moreover, the rpoN mutant markedly reduced extracellular protease and total volatile basic nitrogen (TVB-N) production in sterilized fish juice at 4°C; meanwhile, the juice with the rpoN mutant showed significantly higher sensory scores than that with the wild-type strain. To identify RpoN-controlled genes, RNA-seq-dependent transcriptomics analysis of the wild-type strain and the rpoN mutant was performed. A total of 1224 genes were significantly downregulated, and 474 genes were significantly upregulated by at least two folds at the RNA level in the rpoN mutant compared with the wild-type strain, revealing the involvement of RpoN in several cellular processes, mainly flagellar mobility, adhesion, polysaccharide metabolism, resistance, and amino acid transport and metabolism; this may contribute to the swimming motility, biofilm formation, stress and antibiotic resistance, and spoilage activities of P. fluorescens. Our results provide insights into the regulatory role of RpoN of P. fluorescens in food spoilage, which can be valuable to ensure food quality and safety.

The characteristics of phthalates had been thought to be similar to endocrine disruptors, which increases cancer risk. The role of phthalates in acquired drug resistance remains unclear. In this study, we investigated the effect of di-(2-ethylhexyl) phthalate (DEHP) on acquired drug resistance in breast cancer. MCF7 and MDA-MB-231 breast cancer cells were exposed to long-term physiological concentration of DEHP for more than three months. Long-exposure DEHP permanently attenuated the anti-proliferative effect of doxorubicin with estrogen receptor-independent activity even after withdrawal of DEHP. Long term DEHP exposure significantly reduced ROS (O2−) level in MDA-MB-231 cells while increased in MCF7 cells. ATP-binding cassette (ABC) transporters possess a widely recognized mechanism of drug resistance and are considered a target for drug therapy. Upregulation of ABC family proteins, ABCB-1 and ABCC-1 observed in DEHP-exposed clones compared to doxorubicin-resistant (DoxR) and parental MDA-MB-231 cells. A viability assay showed enhanced multidrug resistance in DEHP-exposed clones against Dox, topotecan, and irinotecan. Inhibition of ABC transporters with tariquidar, enhanced drug cytotoxicity through increased drug accumulation reversing acquired multidrug resistance in MDA-MB-231 breast cancer cells. Tariquidar enhanced Dox cytotoxicity by increasing intracellular ROS production leading to caspase-3 mediated apoptosis. Activation of PI3K/Akt signaling enhanced proliferation and growth of DEHP-exposed MDA-MB-231 cells. Overall, long-term DEHP exposure resulted in acquired multidrug resistance by upregulating ABCB-1 and ABCC1, apart from proliferation PI3K/Akt may be responsible for acquired drug resistance through ABC transporter upregulation. Targeting ABCB1 and ABCC1 with tariquidar may be a promising strategy for reversing the acquired multidrug resistance of triple-negative breast cancer cells.

The clinical treatment of gastric cancer is hampered by the development of anticancer drug resistance as well as the unfavorable pharmacokinetics, nontarget toxicity, and inadequate intratumoral accumulation of current chemotherapies. The polyphenol epigallocatechin gallate in combination with doxorubicin exhibits synergistic inhibition P-glycoprotein efflux pump activity and cancer cell growth. This study evaluated a potential activated nanoparticle delivery system comprising a hyaluronic acid complex with polyethylene glycol-conjugated gelatin containing encapsulated epigallocatechin gallate and low-dose doxorubicin, which may facilitate targeted drug administration to gastric cancer cells. We confirmed successful delivery of bioactive combination drugs and internalization into gastric cancer cells through CD44 ligand recognition and ensuing inhibition of cell proliferation via caspase-induced apoptosis and G2/M phase cell cycle arrest. Furthermore, the targeted nanoparticles significantly suppressed gastric tumor activity and reduced both tumor and heart tissue inflammatory reaction in vivo compared to systemic combination treatment.

Pluronic® F127-modified water-dispersible poly(acrylic acid)-bound iron oxide (PF127-PAAIO) nanoparticles have been prepared as diagnostic agents. A blood–brain-barrier penetrating peptide, angiopep-2 (ANG), was further conjugated onto the surface of the PF127-PAAIO (ANG-PF127-PAAIO) for brain targeting. The ANG-PF127-PAAIO shows negligible cell cytotoxicity, better cellular uptake, and higher T2-weighted image enhancement than the PF127-PAAIO in U87 cells. Using an ex vivo blood–brain barrier (BBB) model, we showed that the ANG-PF127-PAAIO shows better permeability to bypass the BBB. This is because the ANG-PF127-PAAIO has a dual-targeting ability, recognition of the low-density lipoprotein receptor-related protein and clathrin-mediated receptor on the U87 surface. Thus, the ANG-PF127-PAAIO is a potential nanotheranostic agent for brain dysfunction.

Superparamagnetic iron oxide nanoparticles (SPIONs) have been widely used for multiple biomedical applications. Magnetic-assisted transfection (magnetofection) using SPION is an attractive gene vector candidate. In this work, poly(2-dimethylamino)ethyl methacrylate-bound iron oxide nanoparticles (IO-PDMAEMA) were generated using a grafting-from approach via atom transfer radical polymerization (ATRP) for use as a gene vector. Preparing an iron oxide-initiator (IO-initiator) is a typical and important step. We designed a simple method to produce an IO-initiator containing bromide groups (IO-Br). The IO-Br was synthesized by reacting iron oxide nanoparticles with 2-bromoisobutyric acid using a one-pot solvothermal method at a high temperature. We optimized IO-PDMAEMA by controlling the PDMAEMA molecular weight, allowing higher gene expression with lower cytotoxicity. The hydrodynamic diameter of IO-Br was 76.7 nm, which increased to 361.7 nm after polymerization. Transversal relaxivity studies suggested that IO-PDMAEMA can be a contrast agent for magnetic resonance imaging. The magnetofection efficacy of IO-PDMAEMA/pDNA was measured in HEK 293T cells with or without fetal bovine serum (FBS). The IO-PDMAEMA/pDNA magnetoplexes exhibited remarkably improved gene expression in the presence of a magnetic field and 10% FBS compared with a commercial product, PolyMag/pDNA. No significant cytotoxicity of IO-PDMAEMA/pDNA was observed with different incubation time periods with or without the magnetic field. Confocal laser scanning microscopic images showed that the amount of internalized plasmid DNA increased in the assisted magnetic field.

Construction of multifunctional nanoparticles (NPs) with near-infrared (NIR) plasmonic responses is considered a versatile and multifaceted platform for several biomedical applications. Herein, a double layer of Au/Ag alloy on the surface of truncated octahedral iron oxide NPs (IONPs) was prepared and the distance between the layers was controlled to exhibit broad and strong NIR absorption. The rattle-shaped IONP@shell-in-shell nanostructure showed light-response to the NIR biological window from 650 to 1300 nm for photothermal therapy (PTT) and magnetic guidance for hyperthermia and magnetic resonance imaging (MRI) diagnosis. Exposing the aqueous solution of IONP@shell-in-shell to a 1064 nm diode laser, its heat conversion efficiency was ∼28.3%. The in vitro cell viability at a gold concentration of 100 ppm was ∼85%, and decreased to ∼16% when the cells were treated with the NIR irradiation and magnetic attraction. T2-weighted MRI images showed a clear accumulation of IONP@shell-in-shell at the tumor site with magnetic attraction. In vivo luminescence tumor images explained that the IONP@shell-in-shell could reduce the U87MG-luc2 cancer cell proliferation in mice with the NIR irradiation and magnetic attraction. These results indicate the IONP@shell-in-shell as a promising nanomedicine for PTT, magnetic targeting, and magnetic resonance imaging (MRI).

Vacancy and its clusters are among the most important defects induced by self-irradiation in plutonium-gallium (Pu-Ga) alloys. For decades-long stockpiles, the generation and evolution of vacancy defects could cause void swelling and helium bubble formation, resulting in the ageing of the Pu-Ga alloys. Therefore, to shed light on the ageing mechanisms caused by vacancies in Pu-Ga alloys, the long-term behaviour of vacancy defects in Pu-Ga alloys is simulated employing an AKMC model parameterized by molecular statics calculations. By tracking the number of vacancy defects, the size distribution of vacancy clusters, and the largest vacancy clusters over time, we found that temperature and Ga concentration significantly influence the evolution of vacancy defects in Pu-Ga alloys. Temperature changes could affect the clustering behaviour of mono vacancies, and critical temperatures initializing the nucleation of vacancy clusters are observed. On the other hand, adding Ga contents in Pu-Ga alloys could increase the migration energy of and attractive interaction among vacancies, leading to increased vacancy cluster numbers at high temperatures.

Understanding the evolution of irradiation-induced defects is of critical importance for the performance estimation of nuclear materials under irradiation. Hereby, we systematically investigate the influence of He on the evolution of Frenkel pairs and collision cascades in tungsten (W) via using the object kinetic Monte Carlo (OKMC) method. Our findings suggest that the presence of He has significant effect on the evolution of irradiation-induced defects. On the one hand, the presence of He can facilitate the recombination of vacancies and self-interstitial atoms (SIAs) in W. This can be attributed to the formation of immobile He-SIA complexes, which increases the annihilation probability of vacancies and SIAs. On the other hand, due to the high stability and low mobility of He-vacancy complexes, the growth of large vacancy clusters in W is kinetically suppressed by He addition. Specially, in comparison with the injection of collision cascades and He in sequential way at 1223 K, the average sizes of surviving vacancy clusters in W via simultaneous way are smaller, which is in good agreement with previous experimental observations. These results advocate that the impurity with low concentration has significant effect on the evolution of irradiation-induced defects in materials, and contributes to our understanding of W performance under irradiation.

A series of dinuclear aluminum (Al2Pyr2) complexes bridged by two pyrazole ligands were synthesized, and their catalytic activity toward ring-opening polymerization of e-caprolactone (CL) was investigated. Different types of the Al–N–N–Al–N–N skeletal ring were found among these Al2Pyr2 complexes. The butterfly form, LThio2Al2Me4, exerted the highest catalytic activity for CL polymerization. κ2-CL coordination with both Al centers within the butterfly form LThio2Al2Me4 facilitates the initiation process. Generally speaking, the Al2Pyr2 complexes exhibited substantially higher catalytic activity for CL polymerization than literature examples such as β-diketiminate- or traiaza-bearing aluminum complexes. In fact, the Al2Pyr2 complexes can even carry out CL polymerization at room temperature.

A series of five- and six-membered-ring Al complexes bearing Schiff bases was synthesized and their application to the ring-opening polymerization of e-caprolactone (CL) was studied. The five-membered-ring Al complexes have been shown to have a significantly higher polymerization rate than six-membered-ring Al complexes (2–3 fold for CL polymerization). The X-ray data revealed that the Al center of a five-membered-ring Al complex is farther from the Schiff base ligand than is that of a six-membered-ring Al complex. The results of density functional theory calculations also suggest that more space around the Al center of five-membered-ring Al complexes may reduce the steric repulsion in CL polymerization and increase the catalytic activity of five-membered-ring Al complexes.

Carnosic acid is a phenolic diterpene with anti-inflammation, anticancer, anti-bacterial, anti-diabetic, as well as neuroprotective properties, which is generated by many species from Lamiaceae family. Fisetin (3,3',4',7-tetrahydroxyflavone), a naturally flavonoid is abundantly produced in different vegetables and fruits. Fisetin has been reported to have various positive biological effects, including anti-proliferative, anticancer, anti-oxidative and neuroprotective effects. Lung cancer is reported as the most common neoplasm in human world-wide. In the present study, the possible benefits of carnosic acid combined with fisetin on lung cancer in vitro and in vivo was explored. Carnosic acid and fisetin combination led to apoptosis in lung cancer cells. Caspase-3 signaling pathway was promoted in carnosic acid and fisetin co-treatment, which was accompanied by anti-apoptotic proteins of Bcl-2 and Bcl-xl decreasing and pro-apoptotic signals of Bax and Bad increasing. The death receptor (DR) of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) was enhanced in carnosic acid and fisetin combined treatment. Furthermore, the mouse xenograft model in vivo suggested that carnosic acid and fisetin combined treatment inhibited lung cancer growth in comparison to the carnosic acid or fisetin monotherapy. This study supplies a novel therapy to induce apoptosis to inhibit lung cancer through caspase-3 activation.

A series of aluminum (Al) complexes bearing hydrazine-bridging Schiff base and salen ligands were synthesized and investigated as catalysts for the ring-opening polymerization of e-caprolactone (CL). The introduction of steric bulky groups increases the catalytic activity of the corresponding mononuclear aluminum complex. However, the opposite phenomenon was observed in dinuclear Al complexes bearing salen ligands because the steric repulsion reduced the cooperative activation mechanism in the dinuclear Al system. Among these Al complexes, LN2Bu-Al2Me4 bearing a hydrazine-bridging Schiff base ligand had the highest catalytic activity, approximately 3- to 11-fold higher than that of dinuclear Al complexes bearing salen ligands and mononuclear Al complexes bearing Schiff base ligands. Density functional theory calculations revealed that the mechanism of the coordination of CL to one Al center was initiated by the benzyl alkoxide of another Al center.

We report herein the synthesis of a novel photocleavable crosslinker, 4-formylphenyl 4-((4-formylphenoxy)methyl)-3-nitrobenzoate (CHO–ONB–CHO) and its joining with amine-based polysaccharides, viz. chitosan, resulting in the formation of a dual stimuli-responsive (ONB–chitosan) hydrogel having UV- and pH-responsive sites. The detailed mechanism for the formation of CHO–ONB–CHO and ONB–chitosan hydrogel is proposed. The (CHO–ONB–CHO) crosslinker was characterized using 1H-NMR, LCMS and UV-visible spectroscopy. The dual responsive hydrogel is characterized by FTIR, SEM, XRD, DSC and TGA. The crosslinked hydrogel displayed mechanical robustness with a storage modulus of about 1741 pa. The pH-responsiveness of the hydrogel was studied via equilibrium swelling studies in various pH media at 37 °C. The photocleavable behavior of the crosslinker was observed in the UV-absorption range of 310–340 nm and the hydrogel exhibited maximum swelling at pH 5.7. The higher swelling of the hydrogel in acidic conditions and its photo-responsiveness can be exploited for the controlled, temporal and spatial release of therapeutic drugs at any inflammatory areas with acidic environments. It was observed that the hydrogel exhibited higher drug release at pH 5.7 than at pH 7.4.

The functional outcome of intracerebral hemorrhage (ICH) in young male patients are poor than in premenopausal women. After ICH, ferrous iron accumulation causes a higher level of oxidative injury associated with autophagic cell death in striatum of male mice than in females. In rodent model of ferrous citrate (FC)-infusion that simulates iron accumulation after ICH, female endogenous estradiol (E2) suppresses autophagy via estrogen receptor α (ERα) and contributes to less injury severity. Moreover, E2 implantation diminished the FC-induced autophagic cell death and injury in males, whose ERα in the striatum is less than females. Since, no sex difference of ERβ was observed in striatum, we delineated whether ERα and G-protein-coupled estrogen receptor 1 (GPER1) mediate the suppressions of FC-induced autophagy and oxidative injury by E2 in a sex-dimorphic manner. The results showed that the ratio of constitutive GPER1 to ERα in striatum is higher in males than in females. The GPER1 and ERα predominantly mediated suppressive effects of E2 on FC-induced autophagy in males and antioxidant effect of E2 in females, respectively. This finding opens the prospect of a male-specific therapeutic strategy targeting GPER1 for autophagy suppression in patients suffering from iron overload after hemorrhage.

In this manuscript, we have reported a strategy for the synthesis and fine control of the crystal size and morphology of a novel mesoporous faujasite type zeolitic imidazolate framework-8 (ZIF-8) nanocrystals in water solution. The synthesized materials are mesoporous faujasite type, highly crystalline, nanometer-sized, and with a high surface area. The materials have been characterized using powder X-ray diffraction (XRD), transmission electron micrograph (TEM), BET-Surface area, and Fourier transform infrared spectroscopic (FT-IR) analysis. The distribution of nanoparticles is observed through DLS spectroscopy. The thermogravimetry/differential thermal analysis reveals that the material is stable even up to 600 °C. Tetrahedral coordination and 4+ oxidation state of Zinc is found through UV–vis and X-ray photoelectron spectroscopies. The NaBH4 assisted rhodamine B degradation at room temperature shows excellent catalytic activity (∼ 95 %) in a short duration (10 min.). A plausible mechanism of degradation on its catalytic activity is also proposed.

Combining dual drugs in one vehicle to cancer cells offers spatiotemporal localization of drug at the site of action, leading to synergistic therapeutic effects and reduced side effects. To improve pH/redox responsiveness to the tumor microenvironments for cancer therapy, a pH/redox-responsive micelle based on poly(e-caprolactone)-SS-poly(methacrylic acid) (PCL-SS-PMAA) diblock copolymer was fabricated for dual drug delivery. The PCL-SS-PMAA was formulated into a core-shell micelle (PSPm) in an aqueous solution. The critical micelle concentration (CMC) values of PSPm were 7.94 × 10−3 mg mL-1 at pH 5.0 and 1.00 × 10-2 mg mL-1 at pH 7.4. The hydrodynamic diameters of PSPm were within 210–270 nm, depending on pH values. Changes in morphology and size of PSPm were clearly observed before and after exposure to a reducing agent. Paclitaxel (PTX) was encapsulated into the core and cisplatin (CDDP) was chelated on the shell of PSPm, with both PTX and CDDP being efficiently released from PSPm in the presence of a reducing agent in an acid condition. MTT and annexin V/propidium iodide dual staining results demonstrated that co-loading of CDDP and PTX into PSPm had a synergistic effect in killing lung cancer cells and exerted superior antitumor activity over the combination of single drug-loaded PSPm or the combination of free-CDDP and free-PTX at equivalent drug amounts. Hence, encapsulating the dual drugs into PSPm exhibits a synergistic effect for potential lung cancer therapy.

The leaf photosynthesis and nitrogen (N) translocation in three large-spike lines and control cultivar (Xi'nong 979) of winter wheat (Triticum aestivum L.) were studied in 2010–2011 and 2011–2012. The objectives of this study were to investigate the differences in the physiological characteristics of large-spike lines and control cultivar and identify the limiting factors that play a role in improving the yield of breeding materials. The average yield, grain number per spike, kernel weight per spike, and 1000-kernel weight of the large-spike lines were 16.0, 26.8, 42.6, and 15.4%, respectively, significantly higher than those of control. The average photosynthetic rates (Pn) were not significant between the large-spike lines and control cultivar during the active growth period. The average PSII maximum energy conversion efficiency (Fv/Fm), PSII actual quantum efficiency (ΦPS??), photochemical quenching coefficient (qP), PSII reaction center activity (Fv‘/Fv‘) and water-use efficiency (WUE) of the large-spike lines were 1.0, 5.1, 3.6, 0.8, and 43.4%, respectively, higher than those of the control during the active growth stages. The N distribution proportions in different tissues were ranked in the order of grains

A self-assembled poly(methacrylic acid)–poly(e-caprolactone) block copolymer with a disulfide linkage, PMAA-b-PCL-SS-PCL-b-PMAA (S-PCL-PMAA)2, was synthesized for enhanced cellular uptake due to a reduction response to glutathione (GSH) and pH-sensitive characteristics. For comparison, a reduction-insensitive PMAA-b-PCL-CC-PCL-b-PMAA (C-PCL-PMAA)2, using a carbon–carbon linkage as a symmetrical center was also synthesized. These block copolymers were synthesized via the combination of ring opening polymerization (ROP) and atom transfer radical polymerization (ATRP) followed by hydrolysis. The similar number of MAA repeating units was controlled in the copolymers containing either disulfide or carbon–carbon linkages. Copolymers could self-assemble into core–shell micelles in an aqueous solution owing to amphiphilicity. The molecular weight of (S-PCL-PMAA)2 increases linearly with reaction time at both reaction temperatures of 40 and 80 °C. Critical micelle concentrations range within 3.09 × 10−3 to 6.31 × 10−3 mg mL−1 at pH 5 and 3.16 × 10−2 to 3.98 × 10−2 mg mL−1 at pH 8. The average hydrodynamic diameters of micelles are ∼200 nm. The cellular uptake of (S-PCL-PMAA)2 increases with incubation time and is higher in the medium with GSH than without in CRL-5802 cells. In contrast, the cellular uptake of (C-PCL-PMAA)2 is insensitive to the presence of GSH. The higher internalization of the micelle containing disulfides in the presence of GSH is attributable to all three pinocytosis pathways involved, including macropinocytosis-, caveolae-, clathrin-mediated endocytosis, but in the absence of GSH clathrin-mediated endocytosis is only involved. The nascent (S-PCL-PMAA)2 is non-cytotoxic to four cell lines. However, the paclitaxel-encapsulated (S-PCL-PMAA)2 shows slightly higher cell-killing ability than free paclitaxel against CRL-5802 cells. Thus, the GSH-responsive (S-PCL-PMAA)2 is a potential drug delivery system.

We previously synthesized a chondroitin sulfate-graft-poly(ε-caprolactone) copolymer (H-CP) with a high content of poly(ε-caprolactone) (18.7 mol%), which self-assembled in water into a rod-like micelle to encapsulate hydrophobic camptothecin (CPT) in the core (micelle/CPT) for tumor-targeted drug delivery. As a result of the recognition of the micelle by CD44, the micelle/CPT entered CRL-5802 cells efficiently and released CPT efficaciously, resulting in higher tumor suppression than commercial CPT-11. In this study, H1299 cells were found to have a higher CD44 expression than CRL-5802 cells. However, the lower CD44-expressing CRL-5802 cells had a higher percentage of cell death and higher cellular uptake of the micelle/CPT than the higher CD44-expressing H1299 cells. Examination of the internalization pathway of the micelle/CPT in the presence of different endocytic chemical inhibitors showed that the CRL-5802 cells involved clathrin-mediated endocytosis, which was not found in the H1299 cells. Analysis of the cell cycle of the two cell lines exposed to the micelle/CPT revealed that the CRL-5802 cells arrested mainly in the S phase and the H1299 cells arrested mainly in the G2-M phase. A consistent result was also found in the evaluation of γ-H2AX expression, which was about three-fold higher in the CRL-5802 cells than in the H1299 cells. A near-infrared dye, IR780, was encapsulated into the micelle to observe the in vivo biodistribution of the micelle/IR780 in tumor-bearing mice. The CRL-5802 tumor showed a higher fluorescence intensity than the H1299 tumor at any tracing time after 1 h. Thus we tentatively concluded that CRL-5802 cells utilized the clathrin-mediated internalization pathway and arrested in the S phase on exposure to the micelle/CPT; all are possible reasons for the better therapeutic outcome in CRL-5802 cells than in H1299 cells.

Our previous research has confirmed a chondroitin sulfate-polyethylenimine copolymer (CS-PEI) as a potential gene delivery vector because of its recognition by CD44, which enhances the cellular uptake of CS-PEI/pDNA polyplexes. As poly(N,N-dimethylaminoethyl methacrylate) (PDMAEMA) is also a commonly used non-viral gene delivery vector, a CS-PDMAEMA copolymer was synthesized using a similar method with CS-PEI via Michael addition. The physicochemical properties of CS-PDMAEMA and CS-PEI copolymers were thoroughly characterized. The gel electrophoresis results demonstrate that the weight ratio of CS-PEI/pDNA required to completely encapsulate pDNA is ≥1 while that of CS-PDMAEMA/pDNA is ≥3. The CS-modified cationic polymers show lower cytotoxicity than compared with the unmodified ones. At the same weight ratio, CS-PEI/pDNA has a smaller particle size than CS-PDMAEMA/pDNA. The cellular uptake of CS-modified polyplexes is higher in U87 cells (high CD44 expression) than in 3T3 cells (low CD44 expression). However, the transfection efficiency of CS-modified polyplexes is higher in 3T3 cells than in U87 cells. The contrasting results may be attributed to the variation of cell types. In addition, the high level of asialoglycoprotein receptor (ASGP-R) expressed in 3T3 cells seems beneficial for triggering the lectin receptor-mediated endocytosis and results in high transfection efficiency when compared with U87 cells.

Gliomas display a poor disease prognosis causing death within 15 months after diagnosis. Chemotherapy has offered some hope to target this, however, it is majorly ineffective due to the low therapeutic window, poor efficacy and high cytotoxicity. To overcome these challenges, we conjugated Angiopep-2, a cell penetrating peptide (CPP) to Iron Gold (Fe-Au) alloy nanoparticles and investigated the ability of Ang-Fe-Au Nps conjugate to limit glioma growth via magnetic field induced hyperthermia. Our results show that 6.44 nm sized conjugated Fe-Au Nps were superparamagnetic, enhanced negative Glioma image contrast and displayed a 12 °C temperature elevation when magnetically stimulated, indicating applications in medical imaging and hyperthermia-based therapy. Angiopep-2 conjugation resulted in 1.5-fold higher ingestion by C6 glioma cells than L929 fibroblasts, indicating specific glioma targeting and resulting in 90 % decrement in cell viability due to magnetic field induced hyperthermia. Immunohistochemical analysis showed an enhanced coagulative necrosis, glial fibrillary acidic protein (GFAP) expression and decreased Ki67 labelling index in rat treated with Ang-Fe-Au Nps which translated to a 5-fold decrement in tumor volume, consequently resulting in an increased survival time by 7 days. The dual application of this platform opens new doors towards cancer theranostics with minimal invasiveness.

We have developed an object kinetic Monte Carlo (OKMC) code and simulated hydrogen-vacancy clustering behavior and dependence on temperature and hydrogen-vacancy ratio in tungsten. For each of the temperatures we simulated from 300 K to 1000 K, H n V clusters with smaller n form before those with larger n . The elevating temperature leads to a decrease in hydrogen vacancies: H 10 V and H 9 V clusters dominate at 300 K and 600 K, whereas H 5 V , H 6 V , and H 7 V clusters dominate when the temperature reaches 1000 K. Furthermore, only H n V clusters with smaller n formed when a lower hydrogen-vacancy ratio was used due to insufficient availability of hydrogen atoms to occupy vacancies. The results suggest hydrogen emission occurs very rarely at lower temperatures, while higher temperatures facilitate the dissociation of hydrogen from H n V clusters.

The adsorption and dissociation of water monomer and dimer on stepped Co(0001) surface were studied by means of first-principles calculations. Present results indicate that the adsorption strength of water is greatly enhanced by the presence of step, while the activity of water monomer dissociation does not exhibit a noticeable improvement. Nevertheless, water dimer partial dissociation on stepped Co(0001) is more active than on flat Co(0001), and the promotion of oxygen atom on O–H bond cleavage of H2O is more prominent on stepped surface than on flat Co(0001). The findings reveal the importance of low coordinated surface atoms on metallic catalysts and the vital role of surface rippling on water dissociation. Together with previous reports, the activity of water dissociation on cobalt-based catalytic surfaces depends dominantly on O-containing species like oxygen atom, H2O or hydroxyl.

Genetic studies have elucidated mechanisms that regulate aging, however, there has been little progress in identifying drugs that retard ageing. Caenorhabditis elegans is among the classical model organisms in ageing research. Methyl 3,4-dihydroxybenzoate (MDHB) can prolong the life-span of C. elegans, but the underlying molecular mechanisms are not yet fully understood. Here, we report that MDHB prolongs the life-span of C. elegans and delays age-associated declines of physiological processes. Besides, MDHB can lengthen the life-span of eat-2 (ad1113) mutations, revealing that MDHB does not work via caloric restriction (CR). Surprisingly, the life-span&ndash, extending activity of MDHB is completely abolished in daf-2 (e1370) mutations, which suggests that daf-2 is crucial for a MDHB-induced pro-longevity effect in C. elegans. Moreover, MDHB enhances the nuclear localization of daf-16/FoxO, and then modulates the expressions of genes that positively correlate with defenses against stress and longevity in C. elegans. Therefore, our results indicate that MDHB at least partially acts as a modulator of the daf-2/daf-16 pathway to extend the lifespan of C. elegans, and MDHB might be a promising therapeutic agent for age-related diseases.

Background B lymphocyte-induced maturation protein-1 (BLIMP1) is a transcriptional repressor, and plays a crucial role in the regulation of development and functions of various immune cells. Currently, there is limited understanding about the regulation of BLIMP1 expression in keratinocytes and crosstalk between EGFR and BLIMP1 in skin homeostasis. Objective The aim of the study was to investigate the regulation and functional link between EGFR and BLIMP1 in human epidermal keratinocytes. Methods Immunoblotting and Q-PCR were used to determine the molecular mechanism of BLIMP1 expression induced by EGFR in primary human epidermal keratinocytes (NHEK) and HaCaT cells. In functional assay, effects of BLIMP1 knockdown on EGFR-mediated cytokine production, differentiation, and migration in NHEK were evaluated by Q-PCR, ELISA, immunoblotting, and/or wound-healing assay. Results EGFR activation by EGFR ligands could upregulate the protein and mRNA levels of BLIMP1 in NHEK and HaCaT cells. This effect was dependent on PKC, p38, and ERK activation. Additionally, the stability of BLIMP1 protein was under the control of the proteasome and lysosome degradation systems. EGF could also upregulate BLIMP1 expression in skin squamous cell carcinomas. In addition, BLIMP1 knockdown enhanced the EGFR-mediated IL8, CXCL5 and IL6 gene expression and keratinocyte migration, but reduced the EGFR-mediated suppression of differentiation marker K10. Conclusions Our findings shed new insights into the regulation of BLIMP1 expression by EGFR-mediated gene transcription and proteasome/lysosome-mediated degradation in keratinocytes. Functionally, BLIMP1 is a negative regulator of EGF-induced inflammation and migration in keratinocytes, and exerts a gene-specific regulation on keratinocyte differentiation.

We designed a new copolymer, poly(ethylene glycol)-block-poly(e-caprolactone)-graft-poly(acrylic acid) (PAA-PEC), which could be chemically and physically coated onto iron oxide (Fe3O4) nanoparticles for theranostic applications. The chemically PAA-PEC-coated Fe3O4 nanoparticles (PAA-PEC-IO) were prepared using the carboxylic groups of PAA-PEC to bind the Fe3O4 nanoparticles during a co-precipitation reaction. Because of the amphiphilic properties of PAA-PEC, the compound self-assembled into a core-shell structure. The hydrophobic oleic acid-coated Fe3O4 nanoparticles could then be physically encapsulated inside the hydrophobic core of PAA-PEC (PAA-PEC-OA-IO) using an emulsion technique. A similar amount of iron content was controlled in both the PAA-PEC-IO and PAA-PEC-OA-IO (-23%). The particle diameters, morphologies, superparamagnetism, drug loading efficiency, and transversal relaxivity (r2) were studied and compared between the two magnetic nanoparticles. All results displayed the chemically-synthesized PAA-PEC-IO nanoparticles had higher potential than did the physically-synthesized PAA-PEC-OA-IO as an MRI contrast agent and a drug delivery carrier. Rodamine123-linked PAA-PEC-IO (PAA-PEC-IO-Rh123) was used as a molecular probe. Flow cytometric diagrams indicated that cellular internalization of PAA-PEC-IO occurred primarily through clathrin-mediated endocytosis.

We have clarified the performance of two tungsten–helium analytical interatomic potentials, one of which, developed by Li et al., is a bond-order potential, and another, developed by Juslin et al., is a combination of embedded atom method potential and pair potential. Using these two potentials, we have simulated and made a full comparison of formation energy and migration energy of different defects including helium and vacancy, binding energies of helium and vacancy with helium-vacancy cluster, surface energy, as well as melting point, with reference to the corresponding results from the first-principles and experiments.

MicroRNA-128 (miR-128) is an attractive therapeutic molecule with powerful glioblastoma regulation properties. However, miR-128 lacks biological stability and leads to poor delivery efficacy in clinical applications. In our previous study, we demonstrated two effective transgene carriers, including polyethylenimine (PEI)-decorated superparamagnetic iron oxide nanoparticles (SPIONs) as well as chemically-conjugated chondroitin sulfate-PEI copolymers (CPs). In this contribution, we report optimized conditions for coating CPs onto the surfaces of SPIONs, forming CPIOs, for magneto-gene delivery systems. The optimized weight ratio of the CPs and SPIONs is 2 : 1, which resulted in the formation of a stable particle as a good transgene carrier. The hydrodynamic diameter of the CPIOs is ∼136 nm. The gel electrophoresis results demonstrate that the weight ratio of CPIO/DNA required to completely encapsulate pDNA is ≥3. The in vitro tests of CPIO/DNA were done in 293 T, CRL5802, and U87-MG cells in the presence and absence of an external magnetic field. The magnetofection efficiency of CPIO/DNA was measured in the three cell lines with or without fetal bovine serum (FBS). CPIO/DNA exhibited remarkably improved gene expression in the presence of the magnetic field and 10% FBS as compared with a gold non-viral standard, PEI/DNA, and a commercial magnetofection reagent, PolyMag/DNA. In addition, CPIO/DNA showed less cytotoxicity than PEI/DNA and PolyMag/DNA against the three cell lines. The transfection efficiency of the magnetoplex improved significantly with an assisted magnetic field. In miR-128 delivery, a microRNA plate array and fluorescence in situ hybridization were used to demonstrate that CPIO/pMIRNA-128 indeed expresses more miR-128 with the assisted magnetic field than without. In a biodistribution test, CPIO/Cy5-DNA showed higher accumulation at the tumor site where an external magnet is placed nearby.

Chondroitin sulfate-g-poly(ε-caprolactone) (CP) copolymers were synthesized via atom transfer radical addition (ATRA). The CP copolymers self-assembled into micelles in water, and the micelles could be used to encapsulate a hydrophobic anticancer drug, camptothecin (CPT), in the core for tumor targeting delivery. The physicochemical properties of the micelles and CPT-loaded micelles were thoroughly characterized. For the in vitro test, the CPT release, the protection of the lactone ring of CPT from hydrolysis and the cellular uptake of CPT were studied. The cell-killing and apoptosis-inducing effects using the CPT-loaded micelles were significantly better than using free CPT against CRL-5802 cells. The micellar internalization into CRL-5802 cells was primarily via CD44 and clathrin dual-mediated endocytosis. For the in vivo test, the therapeutic efficacy of the CPT-loaded micelles was studied in a non-small-cell lung cancer xenograft animal model. The CPT-loaded micelles showed good inhibition in tumor growth as compared with a commercial product, CPT-11, in CRL-5802 tumor-bearing mice. The in vitro and in vivo data suggested the CP-based micelles are promising anticancer drug vehicles for lung cancer targeting.

A star-shaped poly(e-caprolactone)-b-poly(dimethylaminoethyl methacrylate) copolymer (S-PCL-PDMAEMA) was synthesized and applied to co-deliver pDNA and doxorubicin (DOX) into cancer cells. A linear-shaped L-PCL-PDMAEMA was prepared for comparison. A star-shaped PCL homopolymer (S-PCL) was synthesized through a ring-opening reaction of e-caprolactone with pentaerythritol, followed by brominating the end hydroxyl groups of S-PCL to yield S-PCL-Br. The S-PCL-PDMAEMA was obtained via atom transfer radical polymerization using DMAEMA as a monomer and S-PCL-Br as a macroinitiator. Similar numbers of repeating units of PCL and PDMAEMA were controlled for L-PCL-PDMAEMA and S-PCL-PDMAEMA. The star-shaped copolymer formed uniform nano-sized micelles in water with lower cytotoxicity than the linear one and PDMAEMA. The L-PCL-PDMAEMA and S-PCL-PDMAEMA effectively formed polyplexes with pDNA at a low N/P ratio. The DOX-loaded S-PCL-PDMAEMA micelles showed a better cell-killing effect than the DOX-loaded L-PCL-PDMAEMA in four cell lines. The co-delivery of DOX and pDNA was confirmed using a confocal laser scanning microscope. The S-PCL-PDMAEMA delivered the drugs into the nuclei of U87 cells for 3 h of incubation but the L-PCL-PDMAEMA accumulated most of them in the cytoplasm. This result demonstrated the cationic S-PCL-PDMAEMA micelles are a promising co-delivery system for therapeutic pDNA and hydrophobic anticancer drugs.

The aim of this study was to conjugate an anticancer drug, doxorubicin (DOX) and a folate targeting moiety, folic acid (FA), to self-assembled polycaprolactone (PCL)-graft-chondroitin sulfate (CS) copolymers for enhanced chemotherapy. The PCL-graft-CS copolymer was abbreviated as CP. DOX was conjugated to CP using a bifunctional polyethylene glycol as a spacer (CP-DOX). FA was conjugated to the CP-DOX to yield FA-CP-DOX that could enhance the cellular uptake in folate receptor (FR)-overexpressing cancer cells. The CP-DOX and FA-CP-DOX copolymers were confirmed using 1H-nuclear magnetic resonance (1H-NMR) and Fourier transform infrared (FTIR) spectrophotometers. CP-DOX was spherical and FA-CP-DOX was worm-like. The copolymers without DOX were non-cytotoxic against U87 cells. The IC50 value (an inhibitory concentration of 50% cell growth) of FA-CP-DOX was comparable to that of free DOX but much lower than that of CP-DOX against U87 cells 24, 48 and 72 h post incubation. Because of recognition of the FR, the magnificent cellular uptake of FA-CP-DOX into U87 cells was observed using flow cytometry and confocal laser scanning microscopy.

To evaluate nutrient digestibility, we propose the novel approach of functional group digestibility (FGD). The FGD was based on the absorbance of specific Fourier transform infrared (FT-IR) peaks and the ratio of an inorganic indigestible marker in diet and digesta, without calibration. For application, samples of diet and digesta of wheat with predetermined crude protein (CP) digestibility were scanned on an FT-IR spectrometer equipped with a single-reflection attenuated total reflection (ATR) attachment. The FGD in the amide I region (1689–1631 cm−1) of digesta spectra was strongly related ( R 2 = 0.99) with CP digestibility. The measured diet CP digestibility ranged from 60.4 to 87.8% with a standard error of prediction of 1.09%. In conclusion, instead of predictions based on calibrations, FGD can be calculated directly from spectra, provided the ratio of marker in diet and undigested residue is known, and then accurately predicts nutrient digestibility.

This study investigated the neuroprotective effects of methyl 3,4-dihydroxybenzoate (MDHB) against t-butylhydroperoxide(TBHP) induced oxidative damage in SH-SY5Y (human neuroblastoma cells) and the underlying mechanisms. SH-SY5Y were cultured in DMEM+10% FBS for 24 hours and pretreated with different concentrations of MDHB or N-acetyl-L-cysteine (NAC) for 4 hours prior to the addition of 40 μM TBHP for 24 hours. Cell viability was analyzed using the methyl thiazolyl tetrazolium (MTT) and lactate dehydrogenase (LDH) assays. An annexin V-FITC assay was used to detect cell apoptosis rate. The 2',7'-dichlorofluorescin diacetate (DCFH-DA) assay was used to determine intracellular ROS levels. The activities of antioxidative enzymes (GSH-Px and SOD) were measured using commercially available kits. The oxidative DNA damage marker 8-OHdG was detected using ELISA. Western blotting was used to determine the expression of Bcl-2, Bax, caspase 3, p-Akt and Akt proteins in treated SH-SY5Y cells. Our results showed that MDHB is an effective neuroprotective compound that can mitigate oxidative stress and inhibit apoptosis in SH-SY5Y cells

Men have worse survival than premenopausal women after intracerebral hemorrhage (ICH). After ICH, overproduction of iron associated with induction of heme oxygenase-1 (HO-1) in brain was observed. Rodent ICH model using ferrous citrate (FC)-infusion into the striatum to simulate iron overload, showed a higher degree of injury severity in males than in females. However, the participation of HO-1 in sex-differences of iron-induced brain injury remains unknown. The present results showed a higher level of HO-1 expression associated with more severe injury in males compared with females after FC-infusion. Estradiol (E2) contributed to lower levels of FC-induced HO-1 expression in females compared with males. Heterozygote ho-1 KO decreased the levels of FC-induced injury severity, histological lesions, behavioral deficits, autophagy and autophagic cell death in the striatum of males but not in females. Moreover, ho-1 deficiency enhanced the neuroprotection by E2 only in males. These results suggested that over induction of HO-1 plays a harmful role in FC-induced brain injury in a male-specific manner. Suppression of HO-1 combined with E2 exhibits a synergistic effect on neuroprotection against FC-induced striatal injury in males. These findings open up the prospect for male-specific neuroprotection targeting HO-1 suppression for patients suffering from striatal iron overload.

The aim of this study was to stabilize all-trans-retinol (RE) by complexification with chitosan derivatives through H-bonding. Succinated chitosan (CHI-succ) with three different degrees (5, 10, 20 mol%) of succinylation were synthesized to form complexes with RE. Various weight ratios (w/w) of CHI-succ/RE complexes were prepared and characterized to produce stable complexes in nanometer size. The CHI-succ(0.20)/RE complex with approximate 250 nm in diameter was obtained using a CHI-succ(0.20) concentration of 0.005% (w/v) in double deionized water with various contents of RE. From fine-tuning the degree of succinylation and the weight ratio of the CHI-succ and RE, the formation of supramolecular complexes simultaneously improved water solubility and stability of RE. The cell viability of CHI-succ polymers and their RE complexes in 3T3 cells were all85% relative to the control. The antioxidant ability of the CHI-succ(0.20)/RE complexes was significantly greater than that of pure RE using a 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay (p0.01).

OBJECTIVE To investigate the roles of daidzein in the expressions of steroid receptor coactivator-1 (SRC-1) and nuclear receptor corepressor (NcoR) in MC3T3-E1 osteoblastic cells. METHODS MC3T3-E1 cells were cultured in α-minimal essential medium (α-MEM) containing 2% fetal bovine serum and treated with various concentrations of daidzein (10(-9), 10(-7) and 10(-5) mol/L) or 17β-estradiol at 10(-8) mol/L for 3 d. The protein levels of SRC-1 and NcoR in MC3T3-E1 cells were determined by Western blotting. Estrogen receptor (ER) antagonist ICI182780 at 10(-7) mol/L or specific ERα antagonist methyl-piperidino-pyrazole (MPP) at 10(-6) mol/L were used to block the corresponding receptors, and then MC3T3-E1 cells were treated with daidzein at 10(-7) mol/L or 10(-5) mol/L for 3 d. SRC-1 and NcoR protein levels were detected by Western blotting. RESULTS The protein levels of SRC-1 increased by 2.5 fold (P

It is demonstrated that nanoparticulate PEC with a crosslinked shell sustains DOX release and increases DOX activity against cancer cells. CSMA was synthesized to prepare PEC with chitosan. The double bonds among CSMA were used to form a shell crosslink. The released DOX from DOX-loaded PECs against human cancer KB cells and A549 cells were qualitatively traced by confocal laser scanning microscopy and flow cytometry, and quantitatively measured by capillary electrophoresis. All the results implied the DOX-loaded PEC with a crosslinked shell had the best anti-cancer potency of free DOX and the DOX-loaded PEC prepared from pure chondroitin sulfate and chitosan in both the cell lines.

Folic acid (FA) was selected to link with macromolecules for the selective and specific delivery of doxorubicin (DOX) to folate receptor (FR)-positive tumor cells, because of the high binding affinity of FA to the tumor-associated FR. We synthesized folate-mediated chondroitin sulfate (FA-PEG-ChS) for tumor cell targeting and non-folate-mediated naproxen-linked chondroitin sulfate (Nap-PEG-ChS) for comparison. Both the aforementioned polymers contain a PEG1000 spacer. We encapsulated an anticancer agent, DOX, during the formation of complexes with chitosan. Polyelectrolyte complexes (PEC) grafted with a fluorescent dye (FITC) served as a platform for online imaging cellular internalization. FR-positive KB and FR-deficient A549 cancer cells were tested. The concentration to kill 50% of the cells (IC(50)) of DOX-loaded FA-complex was 1.53 μg/ml, in comparison to 0.91 μg/ml of free DOX. The overlaid fluorescent images of DOX and FITC on confocal laser scanning microscopy demonstrated the co-internalization of DOX and the complex nanoparticles into the cytoplasm of KB cells followed by a gradual release of DOX.

To explore the local mechanisms of fibroblast growth factor (FGF) 23 regulations, we examined the FGF23 expression patterns in an osteoblast culture model. The characteristics of cultured rat calvaria osteoblasts in half-confluence, confluence, osteoid deposition, and osteoid mineralization stages might reflect the proliferation, differentiation, maturation, and matrix mineralization stages, respectively. Compared with proliferating cells in half-confluence, FGF23 expression was upregulated by 7.5-fold at the mRNA level and 126% at the protein level in confluent differentiated cells as determined by real-time RT-PCR and Western blot analysis. Interestingly, mRNA levels of CYP27B1 (the gene coding for 1alpha-hydroxylase enzyme which catalyses the conversion of 1alpha,25-dihydroxyvitamin D, 1alpha,25[OH]2D, from its inactive form, 25-hydroxycholecalciferol, 25[OH]D) and CYP24A (the gene coding for 24-hydroxylase, a target gene of 1alpha,25[OH]2D) were significantly increased by twofold and 34-fold, respectively, in differentiated osteoblasts compared with proliferating cells. We next examined if the local production of 1alpha,25(OH)2D might contribute to the FGF23 upregulation. We cultured osteoblasts in serum-free medium with or without 25-(OH)D (the substrate of 1alpha-hydroxylase). FGF23 mRNA levels were increased in proliferating cells (16-fold) and in differentiated cells (28-fold) by the physiological dose of 25-(OH)D3 treatment. CYP27B1 was slightly but significantly upregulated and CYP24A was increased by 1,700-fold and 800-fold, respectively, in transcriptional levels. Because FGF23 was upregulated in confluent osteoblasts regardless of the presence or absence of 25-(OH)D in serum-free medium, we further examined the possible impact of cell communication on FGF23 expression. We treated osteoblasts with carbenoxolone, a gap junction Cx43 blocker in serum-free medium. The FGF23 mRNA level was reduced by 50% in confluent differentiated cells and slightly but not significantly reduced in half-confluent cells by carbenoxolone treatments. The results suggested that upregulation of FGF23 in differentiated osteoblast appeared to be due to increased autocrine/paracrine action of osteoblast-derived 1alpha,25(OH)2D and increased cell communication, which were identified in cultured rat calvaria osteoblasts. These results indicate that FGF23 expression was stimulated not only by circulating calcitriol but also by locally produced 1alpha,25(OH)2D. The local mechanisms of FGF23 expression remain to be characterized.

Poly(acrylic acid) was decorated onto Fe 3 O 4 resulting in a highly water-soluble superparamagnetic iron oxide. The Poly(acrylic acid) iron oxide (PAAIO) complexes possess specific magnetic properties in the presence of an external magnetic field and are attractive contrast agents for magnetic resonance imaging (MRI). The free carboxylic groups of PAAIO exposed on the surface allow for covalent attachment of a fluorescent dye, Rhodamine 123 (Rh123) to form PAAIO-Rh123, which permits applications in fluorescence imaging. PAAIO-Rh123 is therefore a dual-modality molecular probe. In order to endow specific properties to compounds that target cancer cells and to prevent recognition by the reticuloendothelial system (RES), folic acid-linked poly(ethylene glycol) (FA-PEG) was further conjugated onto PAAIO-Rh123. The amounts of Rh123 and FA-PEG on the modified iron oxides were quantitatively determined by elemental analysis. The iron content was determined by inductively coupled plasma-optical emission spectrometer (ICP-OES). The particle diameters were characterized by dynamic light scattering (DLS) and transmission electron microscope (TEM). Superparamagnetism was confirmed by the superconducting quantum interference device (SQUID) magnetometer. The cellular internalization efficacy of the modified iron oxides was realized in folate-overexpressed FR(+) and folate-deficient FR(−) KB cells by flow cytometry and confocal laser scanning microscopy (CLSM). The quantitative amount of iron internalized into different harvested KB cells was measured by ICP-OES. The T 2 -weighted MR images were tested in FR(+) KB cells.