The Effect of Molecular Structure on Cytotoxicity and Antitumor Activity of PEGylated Nanomedicines.

Fundamental studies on the cellular uptake and drug release of PEGylated nanomedicines are beneficial to understand their fate in vivo and construct ideal nanoparticle formulations. In this work, the detailed metabolic process of PEGylated doxorubicin (Dox) nanomedicines were investigated via confocal laser scanning microscopy (CLSM), flow cytometry (FCM), cytotoxicity test, fluorescence imaging in vivo (FLIV) and liquid chromatography tandem mass spectrometry (LC-MS/MS). Among them, only LC-MS/MS could accurately determine the content of PEGylated Dox and Dox in vitro and in vivo. To the best of our knowledge, this was the first time the PEGylated Dox and released Dox were simultaneously quantified. The interplay of molecular structures, cellular uptake, drug release, and antitumor effect was well characterized. PEG with high molecular weight impeded the cellular uptake of nanoparticles, and the acid-labile hydrazone bond between Dox and PEG promoted Dox release significantly. Cellular uptake and drug release play decisive roles in cytotoxicity and antitumor effect, as evidenced by LC-MS/MS. We emphasized that LC-MS/MS would be a practicable method to quantify PEGylated drugs without complex tags, which could be more in-depth to understand the interaction between PEGylated nanomedicines and their antitumor efficacy.

Expression Characteristics and Functional Analysis of the ScWRKY3 Gene from Sugarcane.

The plant-specific WRKY transcriptional regulatory factors have been proven to play vital roles in plant growth, development, and responses to biotic and abiotic stresses. However, there are few studies on the WRKY gene family in sugarcane (Saccharum spp.). In the present study, the characterization of a new subgroup, IIc WRKY protein ScWRKY3, from a Saccharum hybrid cultivar is reported. The ScWRKY3 protein was localized in the nucleus of Nicotiana benthamiana leaves and showed no transcriptional activation activity and no toxic effects on the yeast strain Y2HGold. An interaction between ScWRKY3 and a reported sugarcane protein ScWRKY4, was confirmed in the nucleus. The ScWRKY3 gene had the highest expression level in sugarcane stem pith. The transcript of ScWRKY3 was stable in the smut-resistant Saccharum hybrid cultivar Yacheng05-179, while it was down-regulated in the smut-susceptible Saccharum hybrid cultivar ROC22 during inoculation with the smut pathogen (Sporisorium scitamineum) at 0⁻72 h. ScWRKY3 was remarkably up-regulated by sodium chloride (NaCl), polyethylene glycol (PEG), and plant hormone abscisic acid (ABA), but it was down-regulated by salicylic acid (SA) and methyl jasmonate (MeJA). Moreover, transient overexpression of the ScWRKY3 gene in N. benthamiana indicated a negative regulation during challenges with the fungal pathogen Fusarium solani var. coeruleum or the bacterial pathogen Ralstonia solanacearum in N. benthamiana. The findings of the present study should accelerate future research on the identification and functional characterization of the WRKY family in sugarcane.

Homeostatic regulation of flavonoid and lignin biosynthesis in phenylpropanoid pathway of transgenic tobacco.

Flavonoids and lignin consist of a large number of secondarymetabolites which are derived from the phenylpropanoid pathway, and they act as a significant role in plant growth, development, and stress response. However, few reports have documented that how different subbranches of phenylpropanoid metablolic pathway mutually interact. In Arabidopsis, AtCPC (AtCAPRICE) is known to play a negative role in anthocyanin accumulation. Nonetheless, whether AtCPC could control the biosynthesis of lignin is largely unknown. Additionally, whether the RrFLS and RrANR, flavonol synthase and anthocyanidin reductase, from Rosa rugosa regulate different branches of phenylpropanoid pathway is unclear. Here, we performed a series of transgenic experiments with short life cycle tobacco and RNA-Seq analysis. Finally, a series of assays related to biological, physiological, and phenotypic characteristics were undertaken. Our results indicated that ectopic expression of AtCPC in tobacco not only decreased the flavonoid compound accumulation, but also up-regulated several lignin biosynthetic genes, and significantly increased the accumulation of lignin. Our results also revealed that although they respectively improved the flavonol and proanthocyanidin contents, the overexpression of RrFLS and RrANR plays positive roles in lignin biosynthesis in transgenic tobacco plants. Our findings provide a novel insight into the mechanism underlying homeostatic regulation of flavonoid and lignin biosynthesis in phenylpropanoid pathway of plants.

Amphiphilic redox-sensitive NIR BODIPY nanoparticles for dual-mode imaging and photothermal therapy.

Theranostics, integrating tumor treatment and diagnosis concurrently, has become an emerging and meaningful strategy in cancer therapy. In this work, an amphiphilic redox-sensitive near-infrared (NIR) BODIPY dye, which could be formed into nanoparticles (PEG-SS-BDP NPs) by self-assembly, was synthesized and possessed good capability of photothermal therapy (PTT), near-infrared fluorescence (NIRF) imaging, photoacoustic (PA) imaging and drug loading. The stable nanoparticles could be dissociated to turn on NIRF due to the rift of embedded disulfide bonds by glutathione (GSH). The enhanced fluorescence in vitro could be observed via confocal laser scanning microscopy (CLSM) after adding GSH, confirming the redox-sensitivity of disulfide bonds. NIRF and PA imaging demonstrated active accumulation in tumor and good imaging effect. At last, PEG-SS-BDP NPs could significantly suppress tumor growth in vivo upon irradiation. The amphiphilic redox-sensitive BODIPY nanoparticles provide a promising design strategy to formulate multifunctional stimuli-responsive theranostic nanoplatforms.

A sensitive amperometric AChE-biosensor for organophosphate pesticides detection based on conjugated polymer and Ag-rGO-NH2 nanocomposite.

Long-term accumulation of organophosphate pesticides in environment presents a potential hazard to human and animal health. Towards this, a highly sensitive amperometric AChE-biosensor based on conjugated polymer and Ag-rGO-NH2 nanocomposite has been successfully developed. First, 4, 7-di (furan-2-yl) benzo thiadiazole (FBThF) was electrochemically polymerized on the electrode surface. Then, Ag-rGO-NH2 nanocomposite and acetylcholinesterase (AChE) are modified on the polymer membrane surface. In this way, a novel amperometric AChE-biosensor was successfully prepared. The as-prepared biosensor possessed excellent conductivity, catalytic activity, and biocompatibility which were attributed to the synergistic effects of poly(FBThF) and Ag-rGO-NH2 and provided a hydrophilic surface for AChE adhesion. Under optimized conductions, the linear range was 0.099-9.9 µg L-1 with a regression coefficient of 0.9947 for malathion, 0.0206-2.06 µg L-1 with a regression coefficient of 0.9969 for trichlorfon. The detection limit is calculated to be about 0.032 µg L-1 for malathion and 0.001 µg L-1 for trichlorfon (S/N = 3). Moreover, the biosensor exhibited acceptable reproducibility and long-term stability, which makes it possible to provide a novel and promising tool for analysis of organophosphate pesticides.

Poly(ε-caprolactone) modified organic dyes nanoparticles for noninvasive long term fluorescence imaging.

Application of organic dyes is limited in biomedical fields due to the rapid self-quenching, poor stability and water solubility. In this work, polymer modified organic dyes is developed through the dyes initiating polymerization. The polymer length is studied by tuning the amount of monomer. With an optimal molecule weight of the polymer, the π-π stacking from π-conjugated organic fluorescent dyes and fluorescence quenching are inhibited, resulting in an enhancement of fluorescence intensity and photostability. Nanoparticles are further fabricated to be used for cell uptake based on the optimized organic dyes. Then, long term cellular fluorescence imaging is realized. This work highlights the potential of polymer modification to improve the performance of organic dyes and expand their applications.

Nifedipine regulated periodontal ligament remodeling: an experimental study in rats.

OBJECTIVE: The aim of this study was to investigate the effects of nifedipine on periodontal ligament remodeling during orthodontic tooth movement. MATERIAL AND METHODS: Sixty male Sprague-Dawley rats were randomly divided into an orthodontic group and groups that received either 10 mg/kg or 40 mg/kg nifedipine (NIF). Immunohistochemical staining and image analysis were used to investigate the expression of matrix metalloproteinase (MMP)-1, -10, -13 and collagen-I in PDL, and maxillary 1st molar displacement was measured. RESULTS: Expression of MMP-1, -10, and -13 was significantly decreased in both NIF groups, while collagen-I expression was markedly increased. NIF significantly inhibited tooth movement. CONCLUSIONS: NIF affects the expression of MMP-1, -10, -13 and collagen-I and tooth movement induced by orthodontic force in rats, thus indicating that calcium channels might be important in mediating PDL remodeling.

Hydrophilic modification and anti-fouling properties of PVDF membrane via in situ nano-particle blending.

Two hydrophilic poly-vinylidene fluoride (PVDF) ultrafiltration membranes were prepared via in situ embedment of nanoparticles (NP), i.e., TiO2 and Al2O3, respectively, and their anti-organic-fouling and anti-biofouling were comprehensively investigated. Characterization of modified PVDF-NP membranes by XRD and FTIR exhibited that nanoparticles were embedded successfully. Series of fast filtration tests demonstrated that in contrary to virgin PVDF membrane, PVDF-NP membranes have high permeability and anti-organic-fouling ability by decreasing the possibility of organic matters deposition and accumulation. Co-existed Ca2+ in feed solution deteriorated the organic fouling in virgin PVDF and PVDF-NP membranes, which was mainly caused by gelation of macromolecular foulants. PVDF-NP membranes were used to form MBR modules for domestic wastewater treatment, and the long-term monitoring evidenced that hydrophilic modified membranes achieved stably high COD and [Formula: see text] rejection efficiencies, and better organic rejection capability than mAO process. PVDF-NP membranes possessed consistently high anti-biofouling ability to maintain stable membrane permeability.

Second Near-Infrared Conjugated Polymer Nanoparticles for Photoacoustic Imaging and Photothermal Therapy.

Photothermal conversion in the second near-infrared (NIR-II) window allows deeper penetration and higher exposure to lasers, but examples of NIR-II photothermal agents are mainly formulated by inorganic compounds. In view of the underlying influence of inorganic materials, a novel NIR-II photothermal nanoagent based on a narrow band gap D-A conjugated polymer (TBDOPV-DT) with 2,2-bithiophene as the donor and thiophene-fused benzodifurandione-based oligo( p-phenylenevinylene) as the acceptor has been developed. More importantly, TBDOPV-DT nanoparticles (TBDOPV-DT NPs) are demonstrated to combine excellent photoacoustic imaging (PAI) and photothermal therapy (PTT) ability. TBDOPV-DT NPs exhibit dramatic photostability and heating reproducibility with a photothermal conversion efficiency of 50%. Especially, the NPs possess a remarkable PTT effect toward cancer cells in vitro and can eliminate tumor cells completely in vivo under 1064 nm laser irradiation, while no appreciable side effects have been observed. This study achieves PAI-guided cancer therapy and sheds light on the future of using organic polymer NPs for the NIR-II PTT of cancer.

Translocation of a Polymer through a Crowded Channel under Electrical Force.

The translocation of a polymer chain through a crowded cylindrical channel is studied using the Langevin dynamics simulations. The influences of the field strength F, the chain length N, and the crowding extent ρ on the translocation time are evaluated, respectively. Scaling relation τ ~ F-α is observed. With the crowding extent ρ increasing, the scaling exponent α becomes large. It is found that, for noncrowded channel, translocation probability drops when the field strength becomes large. However, for high-crowded channel, it is the opposite. Moreover, the translocation time and the average translocation time for all segments both have exponential growth with the crowding extent. The investigation of shape factor 〈δ〉 shows maximum value with increasing of the number of segments outside s. At last, the number of segments inside channel Nin in the process of translocation is calculated and a peak is observed. All the information from the study may benefit protein translocation.

Rational Design of Polymeric Nanoparticles with Tailorable Biomedical Functions for Cancer Therapy.

Polymeric nanoparticles (NPs) play a key role in nanoscale formulations for bioimaging, cancer treatment, and theranostics. In this work, we designed and synthesized a series of hydrophobic polymers (P1-6) with different pendent groups via one-step multicomponent Passerini reaction. These polymers possessed similar molecular structures and various biomedical functions. Interestingly, they could self-assemble into stable NPs in aqueous media. All formed NPs were redox sensitive because of the existence of disulfide bonds in the backbone. The stability of NPs in aqueous media with or without glutathione was systematically evaluated and compared. The optical performance, including fluorescence resonance energy transfer, was characterized under different conditions for those polymers with fluorescent components. Importantly, all formed NPs showed good cytocompatibility toward HeLa cells and different biological functions, including drug loading and delivery, bioimaging with variable fluorescence, and photodynamic activity, as evidenced by experiments in vitro and in vivo. These results demonstrate the great potential of multicomponent reaction to customize versatile polymeric nanoparticles for biomedical applications.

Thiadiazole molecules and poly(ethylene glycol)-block-polylactide self-assembled nanoparticles as effective photothermal agents.

A new photothermal nano-agent was obtained by the coprecipitation of 2,5-Bis(2,5-bis(2-thienyl)-N-dodecyl pyrrole) thieno[3,4-b][1,2,5] thiadiazole (TPT-TT) and a biodegradable amphiphilic block copolymer, methoxypoly(ethylene glycol)2K-block-poly(D,L-lactide)2K (mPEG2K-PDLLA2K). TPT-TT, a donor-acceptor-donor (D-A-D) type small molecule, with bis(2-thienyl)-N-alkylpyrrole (TPT) as the donor and thieno[3,4-b]thiadiazole (TT) as the acceptor was a strong near infrared (NIR) absorber, which could convert the absorbed light energy into heat. The formation of TPT-TT nanoparticles (TPT-NPs), which possessed high stability in water, was confirmed by dynamic light scattering (DLS) and transmission electron microscopy (TEM). TPT-NPs showed high photothermal conversion efficiency (32%) and excellent photostability and heating reproducibility. The photostability of TPT-TT NPs was much better than that of indocyanine green (ICG), a federal drug administration (FDA) approved NIR dye. Besides, TPT-TT NPs exhibited significant photothermal therapeutic effect toward human cervical carcinoma (HeLa) and human liver hepatocellular carcinoma (HepG2) cells, while no appreciable dark cytotoxicity was observed. These results highlight the potential of TPT-TT NPs as an effective photothermal agent for cancer therapy.

Reduction-sensitive amphiphilic copolymers made via multi-component Passerini reaction for drug delivery.

One-step synthesis of amphiphilic polymers containing disulfide bond within the hydrophobic backbone was demonstrated via multi-component Passerini reaction. The obtained polymer was self-assembled into micelles in aqueous solution. Curcumin (CUR), an effective and safe anticancer agent, which was limited by its water insolubility and poor bioavailability, was loaded into the micelles as a model drug. The nanoscale polymeric micelles were confirmed by dynamic light scattering (DLS) and transmission electron microscopy (TEM). Faster intracellular CUR release was observed by confocal laser scanning microscopy (CLSM) in the HeLa cells pretreated with GSH than in the unpretreated ones. Micelles also loaded with NH2-BODIPY which was almost non-fluorescent and gave strong enhanced fluorescence under acid conditions. The phenomenon of the stronger enhanced fluorescence in the pretreated HeLa cells showed further that the obtained polymer was reduction-sensitive. In vitro MTT assays showed that the micelles were biocompatible and CUR-loaded micelles had higher cellular proliferation inhibition in contrast to free CUR toward HeLa cells. These results highlight the potential of using multi-component Passerini reaction to make functional copolymers as smart nanocarriers for drug delivery.

The use of cisplatin-loaded mucoadhesive nanofibers for local chemotherapy of cervical cancers in mice.

Polymer-based local drug delivery system may be suitable for the treatment of cervix cancer. A pilot study was carried out to examine the efficacy of cisplatin-loaded poly(ethylene oxide)/polylactide composite electrospun nanofibers as a local chemotherapy system against cervical cancer in mice via vaginal implantation. The nanofibers were proven to have good mucoadhesive property by in vitro mucoadhesion test and in vivo vaginal retention evaluation. An orthotopic cervical/vaginal cancer model was established by injecting murine cervical cancer U14 cells into the vaginal submucosa nearby the cervix. By inserting the nanofibers mat into the vagina of mice, the cisplatin released from the fiber-mat showed a much more accumulation in the vagina/cervix region than in the peripheral organs such as kidneys, liver, or blood, in contrary to the case of intravenous (i.v) injection. The in vivo trials showed that a better balance between anti-tumor efficacy and systemic safety was achieved in nanofibers group than that in i.v injection group at the equal drug dose. Therefore, electrospun nanofibers present a promising approach to the local drug delivery via vagina against cervical cancer.

Antagonistic effect of protein extracts from Streptococcus sanguinis on pathogenic bacteria and fungi of the oral cavity.

An antibacterial substance from Streptococcus sanguinis (S. sanguinis) is known to have an inhibitory effect on putative periodontal pathogens, but its inhibitory effect on pathogens of oral candidiasis is unknown. In this study, intracellular and exocrine proteins were extracted from S. sanguinis. The antagonistic effect of the protein extracts on Prevotella intermedia (P. intermedia) and Porphyromonas gingivalis (P. gingivalis) was detected by a well-plate technique, and the effects of the protein extracts on biofilms formed by these bacteria were evaluated by confocal laser scanning microscopy. The antagonistic effect of the protein extracts on pathogenic fungi was investigated using Candida albicans (C. albicans) and Candida tropicalis (C. tropicalis). The growth curves of C. albicans and C. tropicalis were determined from ultraviolet absorption measurements, their morphological changes following treatment were observed by optical microscopy and scanning electron microscopy, and the effects of the protein extracts on the thickness of their biofilms and the distribution of dead/live bacteria within the biofilms were detected by confocal laser scanning microscopy. The results showed significant inhibitory effects of the intracellular proteins extracted from S. sanguinis on pathogenic bacteria (P. intermedia and P. gingivalis), fungi (C. albicans and C. tropicalis) and the biofilms formed by them. Furthermore, the growth curves and morphology of C. albicans and C. tropicalis were altered following treatment with the intracellular proteins, resulting in disc-like depressions in the surfaces of the fungal spores and mycelia. By contrast, the exocrine proteins demonstrated no significant inhibitory effect on the pathogenic bacteria, fungi and the biofilms formed by them. Thus, it may be concluded that intracellular proteins of S. sanguinis have antibacterial activity and exert an antagonistic effect on certain pathogenic bacteria and fungi of the oral cavity.

Brain-targeted delivery of trans-activating transcriptor-conjugated magnetic PLGA/lipid nanoparticles.

Magnetic poly (D,L-lactide-co-glycolide) (PLGA)/lipid nanoparticles (MPLs) were fabricated from PLGA, L-α-phosphatidylethanolamine (DOPE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-amino (polyethylene glycol) (DSPE-PEG-NH2), and magnetic nanoparticles (NPs), and then conjugated to trans-activating transcriptor (TAT) peptide. The TAT-MPLs were designed to target the brain by magnetic guidance and TAT conjugation. The drugs hesperidin (HES), naringin (NAR), and glutathione (GSH) were encapsulated in MPLs with drug loading capacity (>10%) and drug encapsulation efficiency (>90%). The therapeutic efficacy of the drug-loaded TAT-MPLs in bEnd.3 cells was compared with that of drug-loaded MPLs. The cells accumulated higher levels of TAT-MPLs than MPLs. In addition, the accumulation of QD-loaded fluorescein isothiocyanate (FITC)-labeled TAT-MPLs in bEnd.3 cells was dose and time dependent. Our results show that TAT-conjugated MPLs may function as an effective drug delivery system that crosses the blood brain barrier to the brain.

[Clinical application of modified distal-jet appliance in distalization of maxillary molar].

OBJECTIVE: To investigate the clinical effects of the maxillary molar distalization with modified Distal-Jet appliance. METHODS: 15 patients (average age 12.3 year-old) with maxillary molars shifting mesially were selected as the subjects. Maxillary molars of all patients were distalized with modified Distal-Jet appliance. RESULTS: All patients achieved good occlusion, normal overjet and overbite with modified Distal-Jet appliance. CONCLUSION: Modified Distal-Jet appliance can distalize maxillary molars effectively.

Heterologous expression and site-directed mutagenesis of endoglucanase CelA from Clostridium thermocellum.

The endoglucanase CelA from Clostridium thermocellum was strongly expressed in Bacillus subtilis. The enzyme was purified by Ni(2+)-affinity chromatography. Site-directed substitution of D278 with an asparagine or an alanine residue surprisingly did not decrease the apparent k(cat) value. Further substitutions of two other potentially critical residues, Y215 and D152, resulted in a 2-fold decrease in apparent k(cat) value for Y215P and complete loss of activity for D152N.