Biomimetic nanovaccine-mediated multivalent IL-15 self-transpresentation (MIST) for potent and safe cancer immunotherapy.

Cytokine therapy, involving interleukin-15 (IL-15), is a promising strategy for cancer immunotherapy. However, clinical application has been limited due to severe toxicity and the relatively low immune response rate, caused by wide distribution of cytokine receptors, systemic immune activation and short half-life of IL-15. Here we show that a biomimetic nanovaccine, developed to co-deliver IL-15 and an antigen/major histocompatibility complex (MHC) selectively targets IL-15 to antigen-specific cytotoxic T lymphocytes (CTL), thereby reducing off-target toxicity. The biomimetic nanovaccine is composed of cytomembrane vesicles, derived from genetically engineered dendritic cells (DC), onto which IL-15/IL-15 receptor α (IL-15Rα), tumor-associated antigenic (TAA) peptide/MHC-I, and relevant costimulatory molecules are simultaneously anchored. We demonstrate that, in contrast to conventional IL-15 therapy, the biomimetic nanovaccine with multivalent IL-15 self-transpresentation (biNV-IL-15) prolonged blood circulation of the cytokine with an 8.2-fold longer half-life than free IL-15 and improved the therapeutic window. This dual targeting strategy allows for spatiotemporal manipulation of therapeutic T cells, elicits broad spectrum antigen-specific T cell responses, and promotes cures in multiple syngeneic tumor models with minimal systemic side effects.

Lipid bilayer-based biological nanoplatforms for sonodynamic cancer therapy.

Sonodynamic therapy (SDT) has been developed as a promising alternative therapeutic modality for cancer treatment, involving the synergetic application of sonosensitizers and low-intensity ultrasound. However, the antitumor efficacy of SDT is significantly limited due to the poor performance of conventional sonosensitizers in vivo and the constrained tumor microenvironment (TME). Recent breakthroughs in lipid bilayer-based nanovesicles (LBBNs), including multifunctional liposomes, exosomes, and isolated cellular membranes, have brought new insights into the advancement of SDT. Despite their distinct sources and preparation methods, the lipid bilayer structure in common allows them to be functionalized in many comparable ways to serve as ideal nanocarriers against challenges arising from the tumor-specific sonosensitizer delivery and the complicated TME. In this review, we provide a comprehensive summary of the recent advances in LBBN-based SDT, with particular attention on how LBBNs can be engineered to improve the delivery efficiency of sonosensitizers and overcome physical, biological, and immune barriers within the TME for enhanced sonodynamic cancer therapy. We anticipate that this review will offer valuable guidance in the construction of LBBN-based nanosonosensitizers and contribute to the development of advanced strategies for next-generation sonodynamic cancer therapy.

Self-adaptive nanoassembly enabling turn-on hypoxia illumination and periphery/center closed-loop tumor eradication.

Solid tumors are regarded as complex evolving systems rather than simple diseases. Self-adaptive synthetic therapeutics are required to cope with the challenges of entire tumors; however, limitations in accurate positioning and destruction of hypoxic niches seriously hinder complete tumor eradication. In this study, we engineer a molecular nanoassembly of sorafenib and a hypoxia-sensitive cyanine probe (CNO) to facilitate periphery/center synergistic cancer therapies. The self-adaptive nanoassembly with cascade drug release features not only effectively kills the peripheral tumor cells in normoxic rims but precisely illuminates hypoxic niches following the reduction of CNO by nitroreductase. More important, CNO is found to synergistically induce tumor ferroptosis with sorafenib via nicotinamide adenine dinucleotide phosphate (NADPH) depletion in hypoxic niches. As expected, the engineered nanoassembly demonstrates self-adaptive hypoxic illumination and periphery/center synergetic tumor eradication in colon and breast cancer BALB/c mouse xenograft models. This study advances turn-on hypoxia illumination and chemo-ferroptosis toward clinical applicability.

Surface-anchored tumor microenvironment-responsive protein nanogel-platelet system for cytosolic delivery of therapeutic protein in the post-surgical cancer treatment.

Nanoparticle-anchored platelet systems hold great potential to act as drug carriers in post-surgical cancer treatment due to their intrinsic ability to target the bleeding sites. However, rational design is still needed to further improve its cargo release profiles to meet the cytosolic delivery of therapeutic proteins with intracellular targets. Herein, we developed a tumor microenvironment (TME)-responsive backpack-conjugated platelet system to enhance intracellular protein delivery, thereby significantly inhibiting tumor recurrence after surgery. Specifically, protein nanogels encapsulating GALA and Granzyme B (GrB) are conjugated on the platelet surface via an acid-sensitive benzoic-imine linker through a biorthogonal reaction (GALA-GNGs-P). Taking advantage of wound-tropism of platelets, GALA-GNGs-P could actively accumulate at the surgical trauma and release nanogels in response to acidic TME for promoting deep penetration. Following cellular uptake, the pore-forming peptide GALA helps nanogels escape from lysosome. Subsequently, high glutathione (GSH) concentration in tumor cytoplasm facilitates GrB release from NGs, leading to intense cell apoptosis. GALA-GNGs-P shows remarkable tumor-targeting capability, high cellular uptake, and outstanding lysosomal escaping ability, which can significantly inhibit tumor recurrence in mice models with incomplete tumor resection. Our findings indicate that platelets bioengineered with TME-responsive protein nanogels provide an option to intracellularly deliver therapeutic proteins for the post-surgical treatment of cancer. STATEMENT OF SIGNIFICANCE: Platelet-based drug delivery systems (DDSs) have gained considerable achievements in post-surgical cancer treatment. However, there is no research exploring their potential in realizing the controllable release of cargoes in the acidic tumor microenvironment (TME). Herein, we developed a TME-responsive bioengineered platelet delivery platform (GALA-GNGs-P) for achieving controllable and effective protein intracellular delivery to overcome post-surgical tumor recurrence. Our surface-anchored nanogel-platelet system has the following advantages: (i) improving the loading efficiency of therapeutic proteins, (ii) affecting no physiological function of platelets, (iii) realizing on-demand cargo release in the acidic TME, and (iv) helping proteins escape from endosomal entrapment. Our findings further explored the prospect of cellular backpack system and realized the controllable release of cargoes in the acidic TME.

Linking the chemical nature of soil organic carbon and biological binding agent in aggregates to soil aggregate stability following biochar amendment in a rice paddy.

Changes in soil aggregation with biochar amendment have been investigated extensively, but how biochar affects the chemical composition of organic carbon (C) and biological binding agents in aggregates and their linkage with soil aggregate stability remains unclear. Soil samples were collected in a rice paddy treated with 0 (C0, control), 10 t ha-1 (C10), 20 t ha-1 (C20) and 40 t ha-1 (C40) biochar for twenty months. The amount and chemical composition of soil organic C (SOC), microbial abundances and glomalin-related soil protein (GRSP) were determined in bulk soil and four fractions: large macroaggregates (>2000 µm), small macroaggregates (250-2000 µm), microaggregates (53-250 µm), and silt + clay (<53 µm). Our results showed that the proportion of >250 µm water-stable aggregates and mean weight diameter were gradually increased with increasing biochar addition rate. The concentrations of SOC, readily oxidizable C and microbial biomass C increased most in the small macroaggregates, followed by microaggregates under biochar amendment. Increasing biochar addition rate gradually decreased the relative contents of alkyl C, O-alkyl C and carbonyl C, but increased those of aromatic C across the aggregates, resulting in a higher aromaticity and hydrophobicity of SOC with respect to the control. The abundances of bacteria, fungi and archaea and the content of GRSP were significantly enhanced in the large and small macroaggregates under the C40 treatment. The proportion of >250 µm aggregates was significantly correlated with the contents of soil organic C fractions, GRSP and microbial abundance. Structural equation modeling further revealed that changes in SOC hydrophobicity and GRSP content under biochar amendment had significant and direct effects on the soil aggregate size distribution. In summary, our findings suggest that biochar amendment in rice paddy could improve soil aggregation through altering the chemical composition of soil organic C and the abundance of biological binding agents.

[Effects of combined application of biochar with organic amendments on enzyme activity and microbial metabolic function of carbon sources in infertile red soil]. / ç”Ÿç‰©è´¨ç‚­é æ–½æœ‰æœºç‰©æ–™å¯¹è´«ç˜ çº¢å£¤é ¶æ´»æ€§å’Œå¾®ç”Ÿç‰©ç¢³æºä»£è°¢åŠŸèƒ½çš„å½±å“.

To improve carbon (C) sequestration and soil fertility of red soil, a two-year (2017 and 2018) field experiment was conducted to investigate the effects of two organic amendments (i.e., corn straw and sheep manure) applied alone or combined with biochar on soil nutrient content, enzyme activities involved in C cycling, and microbial substrate utilization rate in infertile red soil. There were six treatments, including control (non-amendment), corn straw, sheep manure and across biochar treatments (without and with biochar amendment, respectively). The organic amendments and biochar were applied in 2017 and 2018. The results showed that, compared with the control, organic amendments significantly increased soil pH, organic C, total nitrogen, available phosphorus and potassium contents. Compared with straw and manure alone, the biochar co-application with straw or manure significantly increased the contents of soil organic C, available potassium, and available nitrogen, without any significant interactive effects. Application of organic amendments significantly increased the activities of soil ß-glucosidase (BG), cellobiohydrolase (CB), ß-xylosidase (XYL), and peroxidase (PERO). The combined application of biochar and straw significantly reduced the activity of phenol oxidase (PHOX) by 28.6% and PERO by 22.2% in comparison with straw addition alone, respectively, while the combined application of biochar and manure significantly reduced the activities of α-glucosidase (AG) by 46.1%, BG by 50.9%, XYL by 41.6%, and PERO by 31.3% compared with manure addition alone, respectively. Compared with the control, the application of organic amendments significantly enhanced soil basal respiration and microbial utilization rates of carbohydrates, whereas biochar co-application significantly decreased microbial utilization rates of carbohydrates and carboxylic acids. Microbial C source utilization rates were significantly and positively correlated with the activities of BG and PERO. Thus, biochar co-application with organic amendments can enhance nutrient content and reduce enzymatic and microbial metabolic activities, thereby may facilitate C sequestration and fertility of infertile red soil.

Artificial tumor microenvironment regulated by first hemorrhage for enhanced tumor targeting and then occlusion for synergistic bioactivation of hypoxia-sensitive platesomes.

The unique characteristics of the tumor microenvironment (TME) could be exploited to develop antitumor nanomedicine strategies. However, in many cases, the actual therapeutic effect is far from reaching our expectations due to the notable tumor heterogeneity. Given the amplified characteristics of TME regulated by vascular disrupting agents (VDAs), nanomedicines may achieve unexpected improved efficacy. Herein, we fabricate platelet membrane-fusogenic liposomes (PML/DP&PPa), namely "platesomes", which actively load the hypoxia-activated pro-prodrug DMG-PR104A (DP) and physically encapsulate the photosensitizer pyropheophorbide a (PPa). Considering the different stages of tumor vascular collapse and shutdown induced by a VDA combretastatin-A4 phosphate (CA4P), PML/DP&PPa is injected 3 h after intraperitoneal administration of CA4P. First, CA4P-mediated tumor hemorrhage amplifies the enhanced permeation and retention (EPR) effect, and the platesome-biological targeting further promotes the tumor accumulation of PML/DP&PPa. Besides, CA4P-induced vascular occlusion inhibits oxygen supply, followed by photodynamic therapy-caused acute tumor hypoxia. This prolonged extreme hypoxia contributes to the complete activation of DP and then high inhibitory effect on tumor growth and metastasis. Thus, such a combining strategy of artificially-regulated TME and bio-inspired platesomes pronouncedly improves tumor drug delivery and boosts tumor hypoxia-selective activation, and provides a preferable solution to high-efficiency cancer therapy.

[Effects of Nitrogen Fertilizer Management on CH4 and N2O Emissions in Paddy Field].

Methane (CH4) and nitrous oxide (N2O) are two extremely important greenhouse gases in the atmosphere. Nitrogen fertilizer is an important factor affecting CH4 and N2O emissions in rice fields. Rational application of nitrogen fertilizer can not only promote high yields of rice but also reduce greenhouse gas emissions. Existing studies have shown that nitrogen reduction and optimal application can effectively improve the nitrogen use efficiency of rice on the basis of ensuring the yield and reduce the loss of N2O caused by nitrification and denitrification of excessive nitrogen in soil. Fertilization times and fertilizer types have significant effects on CH4 and N2O emissions in paddy fields. In this study, a field experiment was conducted for two consecutive years (2019-2020) to study the effects of fertilizer application on CH4 and N2O emissions from rice fields by setting up four treatments consisting of no fertilizer (CK), customary fertilizer application by farmers (CF), twice fertilizer (TT), and 20% replacement of chemical fertilizer by organic fertilizer (OF) using static chamber-gas chromatography. Additionally, the effect of integrating rice yield and integrated global warming potential (GWP) on the greenhouse gas emission intensity (GHGI) per unit of rice yield was analyzed to explore fertilizer application for yield increase and emission reduction in a typical rice growing area in the middle and lower reaches of Yangtze River. The results showed that:① compared with those of CK, the fertilizer treatments reduced CH4 emissions by 14.6%-25.1% and increased N2O emissions by 610%-1836% in both years; ② compared with those of CF, both the TT and OF treatments showed a trend of increasing CH4 emissions and reducing N2O emissions. CH4 emissions increased by 1.8% (P>0.05) and 14.0% (P<0.05), respectively. The annual average of N2O emissions decreased by 63.3% (P<0.05) and 49.2% (P<0.05) in both the TT and OF treatments, respectively. ③ Compared with that of CK, both fertilizer applications increased rice yield and reduced GHGI; compared with that of CF, the OF and TT treatments increased the average annual rice yield by 17.0% and 10.7%, respectively, and reduced GHGI by 6.8% and 13.7%, respectively. The OF treatment had a better yield increase than that of the TT treatment, and the TT treatment had a slightly better emission reduction than that of the OF treatment. In terms of combined yield and GHG emission reduction, both twice fertilizer (TT) and 20% replacement of chemical fertilizer by organic fertilizer (OF) could reduce the intensity of GHG emission per unit of rice yield and achieve yield increase and emission reduction while ensuring rice yield.

Apoptotic body-mediated intercellular delivery for enhanced drug penetration and whole tumor destruction.

Chemotherapeutic nanomedicines can exploit the neighboring effect to increase tumor penetration. However, the neighboring effect is limited, likely by the consumption of chemotherapeutic agents and resistance of internal hypoxic tumor cells. Here, we first propose and demonstrate that apoptotic bodies (ApoBDs) could carry the remaining drugs to neighboring tumor cells after apoptosis. To enhance the ApoBD-based neighboring effect, we fabricated disulfide-linked prodrug nanoparticles consisting of camptothecin (CPT) and hypoxia-activated prodrug PR104A. CPT kills external normoxic tumor cells to produce ApoBDs, while PR104A remains inactive. The remaining drugs could be effectively delivered into internal tumor cells via ApoBDs. Although CPT exhibits low toxicity to internal hypoxic tumor cells, PR104A could be activated to exert strong cytotoxicity, which further facilitates deep penetration of the remaining drugs. Such a synergic approach could overcome the limitations of the neighboring effect to penetrate deep into solid tumors for whole tumor destruction.

Light-triggered dual-modality drug release of self-assembled prodrug-nanoparticles for synergistic photodynamic and hypoxia-activated therapy.

Photodynamic therapy (PDT) leads to tumor hypoxia which could be utilized for the activation of hypoxia-activated prodrugs (HAPs). However, conventional photosensitizer-loaded nanoformulations suffer from an aggregation-caused quenching (ACQ) effect, which limits the efficiency of PDT and synergistic therapy. Herein, prodrug-nanoparticles (NPs) are prepared by the self-assembly of heterodimeric prodrugs composed of pyropheophorbide a (PPa), hypoxia-activated prodrug PR104A, and a thioether or thioketal linkage. In addition, a novel dual-modality drug release pattern is proposed on the basis of the structural states of prodrug-NPs. Under light irradiation, PR104A is released via photoinduced electron transfer (PET) due to the aggregation state of prodrugs. With the disassembly of prodrug-NPs, the ACQ effect is relieved, and PPa produces singlet oxygen which further promotes the reactive oxygen species (ROS)-sensitive release of PR104A. Such prodrug-NPs turn the disadvantage of the ACQ effect to facilitate drug release, demonstrating high-efficiency synergy in combination with PDT and hypoxia-activated therapy.

Human Lacrimal Production Rates from Modified Schirmer-Tear Test.

SIGNIFICANCE: A simple methodology is presented to quantify basal tear production with a modified Schirmer-tear test. PURPOSE: We introduce a simple clinical procedure to measure quantitative basal tear-production flowrates, QL, from a modified Schirmer-tear test (STT). METHODS: Eight healthy subjects aged at least 18 years underwent modified STTs on both eyes for two visits each. Schirmer strips were sheathed with transparent tape before insertion. Topical anesthetic minimized reflex tearing. Wetting lengths were measured every 30 s for 5 min; QL was calculated from the linear slope of wetting length versus time. Determination of QL requires mass-balance equations on the tear prism and Schirmer strip with strip imbibition kinetics obeying Darcy and Young-Laplace laws. RESULTS: Basal tear production rates varied from essentially 0 to about 2 µl/min. With some exceptions, right and left eyes showed similar tear production rates. CONCLUSIONS: By following the modified STT, QL is established with minimal additional effort over a standard Schirmer test. We predict and observe four different subtypes of imbibition kinetics depending on how short or long the time is for first appearance of the wetting front and on how fast or slow is tear production. For slow lacrimal production rates, the standard 5-min wetting length does not correlate with basal tear production.

Response of microbial community structure and function to short-term biochar amendment in an intensively managed bamboo (Phyllostachys praecox) plantation soil: Effect of particle size and addition rate.

Biochar incorporated into soil has been known to affect soil nutrient availability and act as a habitat for microorganisms, both of which could be related to its particle size. However, little is known about the effect of particle size on soil microbial community structure and function. To investigate short-term soil microbial responses to biochar addition having varying particle sizes and addition rates, we established a laboratory incubation study. Biochar produced via pyrolysis of bamboo was ground into three particle sizes (diameter size<0.05mm (fine), 0.05-1.0mm (medium) and 1.0-2.0mm (coarse)) and amended at rates of 0% (control), 3% and 9% (w/w) in an intensively managed bamboo (Phyllostachys praecox) plantation soil. The results showed that the fine particle biochar resulted in significantly higher soil pH, electrical conductivity (EC), available potassium (K) concentrations than the medium and coarse particle sizes. The fine-sized biochar also induced significantly higher total microbial phospholipid fatty acids (PLFAs) concentrations by 60.28% and 88.94% than the medium and coarse particles regardless of addition rate, respectively. Redundancy analysis suggested that the microbial community structures were largely dependent of particle size, and that improved soil properties were key factors shaping them. The cumulative CO2 emissions from biochar-amended soils were 2-56% lower than the control and sharply decreased with increasing addition rates and particle sizes. Activities of α-glucosidase, ß-glucosidase, ß-xylosidase, N-acetyl-ß-glucosaminidase, peroxidase and dehydrogenase decreased by ranging from 7% to 47% in biochar-amended soils over the control, indicating that biochar addition reduced enzyme activities involved carbon cycling capacity. Our results suggest that biochar addition can affect microbial population abundances, community structure and enzyme activities, that these effects are particle size and rate dependent. The fine particle biochar may additionally produce a better habitat for microorganisms compared to the other particle sizes.

[Effects of bamboo charcoal on the growth of Trifolium repens and soil bacterial community structure].

The effects of addition rates (0, 3% and 9%) and particle sizes (0.05, 0.05-1.0 and 1.0-2.0 mm) of bamboo charcoal on the growth of Trifolium repens and soil microbial community structure were investigated. The results showed that bamboo charcoal addition greatly promoted the early growth of T. repens, with the 9% charcoal addition rate being slightly better than the 3% charcoal addition rate. The effects of different particle sizes of bamboo charcoal on the growth of T. repens were not different significantly. Growth promotion declined with time during 120 days after sowing, and disappeared completely after 5 months. DGGE analysis of the bacterial 16S rDNA V3 fragment indicated that bamboo charcoal altered the soil bacterial community structure. The amount and Shannon diversity index of bacteria in the bamboo charcoal addition treatments increased compared with CK. The quantitative analysis showed that the amount of bacteria in the treatment with bamboo charcoal of fine particle (D < 0.05 mm) at the 9% addition rate was significantly higher than in the other treatments. The fine bamboo charcoal had a great effect on soil bacteria amount compared with the charcoal of other sizes at the same addition rate.