A cascade targeting strategy based on modified bacterial vesicles for enhancing cancer immunotherapy.

BACKGROUND: As an efficient tumor immunotherapy, PD-1 antibody has been gradually used in clinical tumor treatment, but the low response rate and excessive immune response limit its extensive application. RESULTS: Herein, a therapeutic regime for the reinvigoration and activation of the tumor immune microenvironment is introduced to improve the anti-tumor effect of the PD-1 antibody. To comprehensively improve the effect of the immunotherapy and reduce excessive immune response, a biomimetic cascade targeting nanosystem, siRNA@PLOV, which was fused by photothermal sensitive liposomes (PTSLs) and attenuated Salmonella outer membrane vesicles (OMVs), was administered in the tumor therapy for targeting of tumor tissues and T cells within tumor respectively. The fused PLOVs which not only retained the biological character of the OMVs, but also enhanced the drug loading ability. The results demonstrated that the immunogenicity of OMVs and photothermal effects can obviously increase the infiltration of T cells and the silencing of CD38 can effectively improve the T cell cytotoxicity, especially combining with PD-1 antibody. CONCLUSIONS: Interesting, this study revealed that anti-PD-1 administration on the 5th day after siRNA@PLOV treatment had the best performance in killing tumors compared with other groups. In addition, this new therapeutic regime also presents a novel strategy for inducing "vaccine effects", conclusively highlighting its potential in preventing tumor recurrence and improving prognosis.

Conjugates of TAT and folate with DOX-loaded chitosan micelles offer effective intracellular delivery ability.

The key for better antitumor efficacy is to improve the specificity of antitumor drugs for tumor cells and diminish their cytotoxicity to normal tissues. Targeted nanoparticles as antitumor drug delivery system can resolve this problem. In this study, we prepared folate and TAT (arginine-rich cell-penetrating peptide) modified N-PEG-N'-octyl-chitosan to form the folate/TAT-PEG-OC micelles. Then, the molecular structure, morphology, size distribution and bio-safety of the micelles were characterized. In order to investigate the drug-loading capacity of folate/TAT-PEG-OC micelles, doxorubicin (DOX) was used as model drug to prepare DOX-loaded chitosan micelles. Here, the confocal microscopy was used to evaluate the cellular uptake of DOX/folate/TAT-PEG-OC micelles, while the self-built NIR imaging system was used to evaluate the dynamic behavior of ICG-Der-01/folate/TAT-PEG-OC micelles in vivo. Our results demonstrate that the dual-modified PEG-OC micelles not only have good morphology, uniform size distribution and excellent drug loading capacity, but also show a strong capability for the efficient intracellular uptake and enhanced targeting behaviors in a folate receptor positive tumor model (Bel-7402 human hepatocellular cells). All these results suggest the potential application of folate/TAT-PEG-OC micelles in the targeted diagnosis and therapy to different kinds of folate receptor positive tumors.

Highly luminescent water-soluble quaternary Zn-Ag-In-S quantum dots for tumor cell-targeted imaging.

Exploring the synthesis and biomedical applications of biocompatible quantum dots (QDs) is currently one of the fastest growing fields of nanotechnology. Hence, in this work, we present a facile approach to produce water-soluble (cadmium-free) quaternary Zn-Ag-In-S (ZAIS) QDs. Their efficient photoluminescence (PL) emissions can be tuned widely in the range of 525-625 nm by controlling the size and composition of the QDs with the PL quantum yields (QYs) of 15-30%. These highly luminescent ZAIS QDs are less toxic due to the absence of highly toxic cadmium, and can be versatilely modified by a DHLA-PEG-based ligand. Importantly, after being modified by tumor cell-specific targeting ligands (e.g., folate and RGD peptide), the PEGylated quaternary QDs show potential applications in tumor cell imaging as a promising alternative for Cd-based QDs.

CO2 emission accounting and emission reduction analysis of the steel production process based on the material-energy-carbon correlation effect.

This paper develops a process-level carbon emission calculation model for iron and steel enterprises through the carbon emission mechanism of the whole production process. The relationship between material, energy and carbon flows is considered and combined. The carbon emissions of enterprises are divided into industrial emissions and combustion emissions, and the indirect emissions of purchased intermediate products and electricity purchased from the grid are also considered. Carbon emission targets and corresponding emission reduction strategies are formulated at the enterprise and process levels. For example, consider an iron and steel enterprise. The different types of carbon emissions are accounted for, with their reduction potential analysed based on the carbon material flow analysis method. The results show that the carbon emission of this enterprise is 1930.87 kgCO2/t (CS), and the combustion emission caused by energy flow is the main contributor to the enterprise's carbon emission, accounting for 57.02% of the total emission. The carbon emission during iron-making accounts for 69.06% of the entire process and is critical in any carbon emission reduction of the enterprise. Among them, process emissions from the blast furnace process account for 81.79% of industrial emissions of the whole process, which is 356.51 kgCO2/t (CS), and is the main challenge of low carbon transformation in this extensive process. This study highlights that increasing the integrated steel-making scrap ratio and electric furnace steel production can break through the existing emission reduction limits. A 65.02% carbon emission reduction can be achieved, and using green electricity can reduce emissions by 24.15%. Reasonably determining the amount of purchased coke and paying attention to the high-value recycling of byproduct gas resources in the plant are essential to achieve low-carbon economic development of steel.

In vivo anti-tumor efficacy of docetaxel-loaded thermally responsive nanohydrogel.

Thermally responsive poly(N-isopropylacrylamide-co-acrylamide) (P(NIPA-co-AAm)) nanohydrogel (NHG) with a diameter of about 50 nm and a lower critical solution temperature (LCST) of about 40 degrees C was synthesized by a previously reported precipitation polymerization method. The physical properties including LCST, diameter and morphology were characterized. Four hydrophobic model drugs (5-fluorouracil (5-FU), fluorescein, docetaxel (DTX) and near-infrared dye-12 (NIRD-12)) with different hydrophilicities were respectively entrapped into the nanoparticles and their in vitro release kinetics from NHG was investigated. DTX was ultimately chosen as the goal anti-tumor drug and optimally entrapped into NHG with a drug loading content (DLC) of 7.38% and encapsulation efficiency (EE) of 73.8%. An in vitro drug release test indicated that DTX-loaded NHG had zero-order release kinetics at 43 degrees C. The respective anti-tumor efficacy of DTX-loaded NHG with or without hyperthermia on tumor tissue was evaluated in Kunming mice-bearing S180 sarcoma. The inhibition rates of DTX-loaded NHG with or without hyperthermia were 78.15% and 48.78%, respectively. DTX-loaded NHG also showed much lower toxicity during the therapeutic procedure. Results indicated that this kind of thermally responsive, drug-loaded NHG could be used as a promising strategy for tumor therapy with the help of local hyperthermia treatment.

[Fabrication, characterization and drug release characteristics of drug loaded poly (L-lactic acid) fiber].

To develop a long-term released and implantable biodegradable fiber carrier for hydrophobic drug incorporation, fibers were fabricated by organic phase separation method. The structure of the fiber was observed by scanning electron microscopy and the state of the drug dispersed in the polymer was measured by differential scanning colorimetry (DSC) and Fouvier transform infrared spectroscopy (FTIR). The drug loading content and release profiles were determined by high performance liquid chromatography (HPLC) and ultra-violet spectrophotometry, respectively. Blank and drug loaded fibers were successfully fabricated and the drug was entirely encapsulated into the fiber. The drug was dispersed in the polymer with minicrystal and noncrystal form. The drug release profile was long-term sustained and could be regulated. The PLLA fibers in micrometer range successfully fabricated by organic phase separation method could be taken as sustained release and implantable drug carrier.

Characterization of pH- and temperature-sensitive hydrogel nanoparticles for controlled drug release.

Monodispersed poly (N-isopropylacrylamide) (PNIPA) nanoparticles and their derivative poly (N-isopropylacrylamide-co-acrylic acid) (PNIPA-co-AA) nanoparticles were successfully synthesized. Regulation of the size and the lower critical solution temperature (LCST) of the hydrogel nanoparticles was intensively studied. The results showed that the diameters and LCST of the particles can be arbitrarily manipulated according to different application designs. The relationship between the size of the particles and the amount of surfactant were quantitatively disclosed. It was found that the LCST of PNIPA-co-AA nanoparticles ranging from 35-45 degrees C correlated with the molar fraction of acrylic acid which was copolymerized with NIPA. The pH sensitivity of PNIPA-co-AA nanoparticles was displayed by the transmittance transition of their aqueous solution in various pH conditions. Furthermore, the anti-cancer drug 5-fluorouracil (5-Fu) was first loaded into both PNIPA and PNIPA-co-AA hydrogel nanoparticles with an entrapment efficiency larger than 4%. In vitro release of 5-Fu from PNIPA-co-AA nanoparticle hydrogels was shown to be pH- and temperature-dependent, which demonstrated that the PNIPA-co-AA nanoparticles have great potential in the design of controlled drug delivery systems.

Cypate-mediated thermosensitive nanoliposome for tumor imaging and photothermal triggered drug release.

It is an emerging focus to explore controlled release drug delivery systems for simultaneous cancer imaging and therapy. Herein, we synthesized a photothermal sensitive multifunctional nano-liposome drug delivery system, with doxorubicin wrapped in the hydropholic layer as the therapeutical agent and cypate doped in the hydrophobic layer as the diagnostic agent. A series of in vitro and in vivo characterization demonstrated the stability of synthesized liposome, as the DL% was 9 ± 1.5 and the EE% was 82.7 ± 2.1. And the liposome achieved the functions of target-delivery, enhanced photochemical internalized drug release, and simultaneous chemotherapy and thermal therapy, indicating that this multifunctional nano-liposome is a promising drug delivery system for tumor diagnosis and targeting therapy.

Photodynamic Therapy Induced Enhancement of Tumor Vasculature Permeability Using an Upconversion Nanoconstruct for Improved Intratumoral Nanoparticle Delivery in Deep Tissues.

Photodynamic therapy (PDT) has recently emerged as an approach to enhance intratumoral accumulation of nanoparticles. However, conventional PDT is greatly limited by the inability of the excitation light to sufficiently penetrate tissue, rendering PDT ineffective in the relatively deep tumors. To address this limitation, we developed a novel PDT platform and reported for the first time the effect of deep-tissue PDT on nanoparticle uptake in tumors. This platform employed c(RGDyK)-conjugated upconversion nanoparticles (UCNPs), which facilitate active targeting of the nanoconstruct to tumor vasculature and achieve the deep-tissue photosensitizer activation by NIR light irradiation. Results indicated that our PDT system efficiently enhanced intratumoral uptake of different nanoparticles in a deep-seated tumor model. The optimal light dose for deep-tissue PDT (34 mW/cm(2)) was determined and the most robust permeability enhancement was achieved by administering the nanoparticles within 15 minutes following PDT treatment. Further, a two-step treatment strategy was developed and validated featuring the capability of improving the therapeutic efficacy of Doxil while simultaneously reducing its cardiotoxicity. This two-step treatment resulted in a tumor inhibition rate of 79% compared with 56% after Doxil treatment alone. These findings provide evidence in support of the clinical application of deep-tissue PDT for enhanced nano-drug delivery.

MUC1 aptamer-based near-infrared fluorescence probes for tumor imaging.

PURPOSE: DNA aptamer (APT) is able to bind to Mucin 1 (MUC1) specifically. The possibility of APT acting as a moiety to construct tumor-targeting probes was investigated. PROCEDURES: A near-infrared (NIR) fluorescent dye (MPA) and polyethylene glycol (PEG) were conjugated to APT to form APT-MPA and APT-PEG-MPA. The successful synthesis of the two probes was characterized via thin layer chromatography (TLC) and optical spectra. The tumor-targeting efficacy of the probes was evaluated in detail at cell level and animal level, respectively. RESULTS: The results indicated that MPA and PEG were successfully coupled with APT. APT-based probes were mediated by Mucin 1 into tumor cells, and PEG-modified probe exhibited higher cell affinity. CONCLUSIONS: The aptamer-based NIR fluorescent probes are promising candidates for tumor imaging and diagnosis.

Thermal responsive micelles for dual tumor-targeting imaging and therapy.

Two kinds of thermally responsive polymers P(FAA-NIPA-co-AAm-co-ODA) and P(FPA-NIPA-co-AAm-co-ODA) containing folate, isopropyl acrylamide and octadecyl acrylate were fabricated through free radical random copolymerization for targeted drug delivery. Then the micelles formed in aqueous solution by self-assembly and were characterized in terms of particle size, lower critical solution temperature (LCST) and a variety of optical spectra. MTT assays demonstrated the low cytotoxicity of the control micelle and drug-loaded micelle on A549 cells and Bel 7402 cells. Then fluorescein and cypate were used as model drugs to optimize the constituents of micelles for drug entrapment efficiency and investigate the release kinetics of micelles in vitro. The FA and thermal co-mediated tumor-targeting efficiency of the two kinds of micelles were verified and compared in detail at cell level and animal level, respectively. These results indicated that the dual-targeting micelles are promising drug delivery systems for tumor-targeting therapy.

In vivo NIR imaging with PbS quantum dots entrapped in biodegradable micelles.

In this article, we firstly synthesized oil-soluble PbS quantum dots (QDs) emitting in the near-infrared (NIR) spectral range through a two-phase method, which exhibit a conveniently tunable photoluminescence (PL) emission (from ~750 to 872 nm) with a narrow PL bandwidth, as well as a high (up to 40%) PL quantum yield (QY). Next, the as-prepared oil-soluble NIR PbS QDs were applied to the in vivo imaging of tumors by entrapping in biodegradable micelles (N-succinyl-N'-octyl nanomicelles, SOC) which had hydrophobic inner cores. Transmission electron microscope results show well dispersed spherical shaped QDs-loaded SOC micelles with 100 nm diameter. The QY of PbS QDs entrapped into SOC has no significant change compared to free QDs. Besides, both in vitro and in vivo acute toxicity results demonstrated that the QDs-loaded micelles have low cytotoxicity. Furthermore, in vivo imaging of PbS QDs-loaded SOC injected intravenously into tumor-bearing nude mice showed the NIR QDs-loaded micelles can accumulate in the tumor tissue due to the enhanced permeability and retention effect of SOC micelles. These results confirm that the as-prepared PbS QDs could be used as fluorescence probes to study the biodistribution of nanocarriers and their intracellular pathways, as well as their passive targeted behavior to tumors in preclinical research.

Folate-polyethylene glycol conjugated near-infrared fluorescence probe with high targeting affinity and sensitivity for in vivo early tumor diagnosis.

PURPOSE: The purpose of this study is to synthesize a folate-polyethylene glycol (PEG) conjugated near-infrared fluorescence probe (fPI-01) for diagnosis of folate receptor (FR)-overexpressed tumors with high sensitivity and specificity. PROCEDURES: fPI-01 was synthesized, purified, and characterized. Its cytotoxicity and affinity to tumor cells were determined in vitro. The dynamics and biodistribution of the probe was monitored in normal nude mice. And the tumor-targeting capability was investigated in nude mice bearing different tumor xenograft. RESULTS: fPI-01 was successfully synthesized with strengthened optical properties. Cells experiments showed the probe had high FR affinity and without apparent cytotoxicity. Animal experiments indicated the probe excreted through urine by kidney. And its tumor-targeting ability was demonstrated on different tumor-bearing mice, with high sensitivity and tumor-to-normal tissue contrast ratio (10:1). CONCLUSIONS: fPI-01 is a promising optical agent for diagnosis of FR-positive tumors, especially in their early stage.

Comparison of two polymeric carrier formulations for controlled release of hydrophilic and hydrophobic drugs.

Two temperature sensitive drug carriers, poly (N-isopropylacrylamide-co-acrylic acid) (PNIPA-co-AA) and poly (N-isopropylacrylamide-vinyl pyrrolidone-acrylic acid) (PNIPA-VP-AA), were successfully synthesized through free radical mechanism. The diameters of PNIPA-co-AA and PNIPA-VP-AA particles can be regulated to be less than 100 nm, which were related to surfactant sodiumdodecyl sulfate and initiator ferrous ammonium sulfate, respectively. The lower critical solution temperature (LCST) of them can be manipulated to be higher than 40 degrees C, which was correlated to amount of acrylic acid (AA) that was copolymerized with NIPA. Hydrophilic anti-tumor drugs, 5-fluorouracil (5-Fu) and hydrophobic drug thalidomide were entrapped into PNIPA-co-AA and PNIPA-VP-AA, respectively. For different interaction mechanism between drug and carrier, 5-Fu was prone to be entrapped in PNIPA-co-AA with loading efficiency larger than 10% (w/w), while thalidomide was entrapped in PNIPA-VP-AA up to 80% (w/w). Fluorescein, an angiography agent, was used to evaluate the drug loading mechanism between PNIPA-VP-AA and poor water-soluble drug. In vitro drug release behavior from these two drug carriers were significantly different and showed temperature dependent, which demonstrated that PNIPA-co-AA and PNIPA-VP-AA are promising candidates for different controlled drug delivery system.

The targeted behavior of thermally responsive nanohydrogel evaluated by NIR system in mouse model.

Thermally responsive poly(N-isopropylacrylamide-co-acrylamide) (P(NIPA-co-AAm)) nanohydrogels were synthesized in this study by free-radical precipitation polymerization method. Different lower critical solution temperatures (LCST) of nanohydrogels were obtained by modulating the amount of AAm and characterized by measuring their transmittances of the particle solutions at 500 nm. The diameters within the range of 50-450 nm were achieved by manipulating the amount of sodiumdodecyl sulfate (SDS). Near infrared dye NIRD-12, with excitation and emission wavelengths at 772 nm and 814 nm, was entrapped into the P(NIPA-co-AAm) nanohydrogels for in vivo animal study. The thermally targeted behavior of the nanohydrogels was evaluated by the in vivo fluorescence imaging on different groups of denuded mice, with or without S180 tumors and with or without hyperthermia treatment. Results indicated that this kind of thermally responsive nanohydrogel could only accumulate in the tissue with higher temperature, no matter normal tissue and tumor site. The thermally targeted behavior is passive and non-specific. The targeted location can be selected by hyperthermia treatment and manipulating the suitable LCST of nanohydrogel. Results indicated that the thermally responsive P(NIPA-co-AAm) nanohydrogel could be used as an attracting thermally targeted carrier for drugs, especially for anti-cancer drugs.

The implantable 5-fluorouracil-loaded poly(l-lactic acid) fibers prepared by wet-spinning from suspension.

The paper introduced an improved fabrication technique, by which the hydrophobic polymer monofilament fiber loading hydrophilic drug was obtained. The micronized 5-Fu (5-fluorouracil) powders were homogeneously dispersed in PLLA (poly(l-lactic acid))-chloroform solution to form the suspension, and then the suspension was solidified in the nonsolvent to prepare the fibers by wet-spinning method under mild condition. The diameter of drug-loaded fiber was in the range of 50-250 microm. The hydrophilic drug powders were successfully encapsulated into the monofilament fiber with good stability, high drug loading content and efficacy. The MTT cytotoxicity assay in vitro revealed the satisfactory anticancer activity of the drug-loaded fibers. The long-term release characteristics of these fibers were also achieved. Furthermore, the drug release rate of the fibers could be regulated by the formulation and fabricating parameters, such as drug loading content, polymer concentration in suspension, nonsolvent composition and flow rate in wet-spinning. The release mechanism of the fibers was investigated and described by Fickian diffusion equation.