Hitchhiking of Cas9 with nucleus-localized proteins impairs its controllability and leads to efficient genome editing of NLS-free Cas9.

CRISPR-Cas9 is the most commonly used genome-editing tool in eukaryotic cells. To modulate Cas9 entry into the nucleus to enable control of genome editing, we constructed a light-controlled CRISPR-Cas9 system to control exposure of the Cas9 protein nuclear localization signal (NLS). Although blue-light irradiation was found to effectively control the entry of Cas9 protein into the nucleus with confocal microscopy observation, effective gene editing occurred in controls with next-generation sequencing analysis. To further clarify this phenomenon, a CRISPR-Cas9 editing system without the NLS and a CRISPR-Cas9 editing system containing a nuclear export signal were also constructed. Interestingly, both Cas9 proteins could achieve effective editing of target sites with significantly reduced off-target effects. Thus, we speculated that other factors might mediate Cas9 entry into the nucleus. However, NLS-free Cas9 was found to produce effective target gene editing even following inhibition of cell mitosis to prevent nuclear import caused by nuclear membrane disassembly. Furthermore, multiple nucleus-localized proteins were found to interact with Cas9, which could mediate the "hitchhiking" of NLS-free Cas9 into the nucleus. These findings will inform future attempts to construct controllable gene-editing systems and provide new insights into the evolution of the nucleus and compatible protein functions.

Impact of COVID-19 restrictions liberalization on air quality: a case study of Chongqing, Southwest China.

To mitigate the societal impact of the COVID-19 pandemic, China implemented long-term restrictive measures. The sudden liberalization at the end of 2022 disrupted residents' daily routines, making it scientifically intriguing to explore its effect on air quality. Taking Chongqing City in Southwest China as an example, we examined the impact of restriction liberalization on air quality, identified potential sources of pollutants, simulated the effects of abrupt anthropogenic control relaxation using a Random Forest Model, and applied an optimized model to predict the post-liberalization pollutant concentrations. The results showed increases in PM2.5 (72.3%), PM10 (67.7%), and NO2 (21.9%) concentrations, while O3 concentration decreased by 20.5%. Although potential pollution source areas contracted, pollution levels intensified with northeastern Sichuan, interior Chongqing, and northern Guizhou being major contributors to pollutant emissions. Anthropogenic emissions accounted for 26.7 ~ 33% changes in PM2.5 and PM10 concentrations while meteorological conditions contributed to 40.2 ~ 43.3% variations observed during the period. The optimized model demonstrated a correlation between predicted and observed values with R2 ranging from 0.70 to 0.89, enabling accurate prediction of post-liberalization pollutant concentrations. This study can enhance our understanding regarding the impact of sudden social lockdown relaxation events on air quality while providing support for urban air pollution prevention.

Single-injection subunit vaccine for rabies prevention using lentinan as adjuvant.

Current rabies vaccines require 5 doses to provide full protection from the deadly virus, which significantly reduce the compliance of recipients. To minimize the number of immunizations herein single injection vaccines were developed. First a single injection vaccine was designed using rabies virus glycoprotein (G protein) as antigen. A time-controlled release system which uses dynamic layer-by-layer films as erodible coating was employed to accomplish multiply pulsatile releases of G protein. The single-injection vaccine elicits potent humoral and cellular immune responses comparable to the corresponding multi-dose ordinary vaccines because of their similar release pattern of G protein. To further improve its performance, a second single injection vaccine, in which lentinan was added as adjuvant, was designed. This single-injection vaccine again elicits humoral and cellular immune responses comparable to the corresponding multi-dose ordinary vaccines because of their similar release pattern of antigen and adjuvant. In addition, the second single-injection vaccine elicits higher level immune response and provides higher efficiency on virus inhibition than the first one because lentinan can booster immune response.

Inflammation-responsive PCL/gelatin microfiber scaffold with sustained nitric oxide generation and heparin release for blood-contacting implants.

Delayed endothelialization, the excessive proliferation of smooth muscle cells (SMCs), and persistent inflammation are the main reasons for the implantation failure of blood-contacting materials. To overcome this problem, an inflammation-responsive, core-shell structured microfiber scaffold is developed using polycaprolactone (PCL), selenocystamine-modified gelatin (Gel-Se), L-ascorbyl 6-palmitate (AP), and dexamethasone as the fiber shell, with poly (l-lysine) (PLL) and heparin incorporated in the fiber core. Superhydrophilic microfiber scaffolds exhibit antifouling properties that inhibit protein adsorption and blood cell adhesion, thereby effectively mitigating the risk of acute thrombosis. The continuous release of heparin and sustained generation of nitric oxide (NO) through the catalytic decomposition of S-nitrosothiols by selenocystamine lead to a biomimetic endothelial function for the enhancement of blood compatibility. The inflammation-responsive compound AP can detoxify excess reactive oxygen species (ROS) while controlling the release of dexamethasone to reduce chronic inflammation. We demonstrate the ability of microfiber scaffolds to reduce thrombotic and inflammatory complications, inhibit SMC proliferation, and promote rapid endothelialization both in vitro and ex vivo. Hence, microfiber scaffolds are robust and promising for blood-contacting implants with enhanced antithrombogenicity and anti-inflammatory capabilities.

Genome Editing of Mammalian Cells Through RNA Transcript-Mediated Homologous Recombination Repair.

Double-stranded break (DSB) repair of eukaryotic DNA is mainly accomplished by nonhomologous end joining and homologous recombination (HR). Providing exogenous templates during HR repair can result in the editing of target genes, which is the central mechanism of the well-established clustered regularly interspaced short palindromic repeats (CRISPR) gene editing system. Currently, exogenous templates are mainly DNA molecules, which can provoke a cellular immune response within the cell. In order to verify the feasibility of RNA molecules as repair templates for HR in mammalian cell genome editing, we fused RNA template molecules to the 3'-end of single guide RNA (sgRNA), so that the sgRNA and the homologous template RNA form a single RNA molecule. The results show this construct can be used as a repair template to achieve target gene editing in mammalian cells. In addition, the factors influencing HR mediated by RNA template molecules were investigated, and it was found that increasing the length of homologous arms and inducing an R-loop near the DSBcan effectively promote HR repair. Furthermore, intracellular homologous chromosomes may compete with exogenous RNA templates. The findings in this article provide a reference for the utilization of RNA template molecules to mediate target gene editing in eukaryotic cells, as well as a basis for the study of the mechanism by which RNA molecules mediate the repair of DSBs.

Heparinized self-healing polymer coating with inflammation modulation for blood-contacting biomedical devices.

The current techniques for antithrombotic coating on blood-contacting biomedical materials and devices are usually complex and lack practical feasibility with weak coating stability and low heparin immobilization. Here, a heparinized self-healing polymer coating with inflammation modulation is introduced through thermal-initiated radical copolymerization of methacrylate esterified heparin (MA-heparin) with methyl methacrylate (MMA) and n-butyl acrylate (nBA), followed by the anchoring of reactive oxygen species (ROS)-responsive polyoxalate containing vanillyl alcohol (PVAX) onto the coating through esterification. The aspirin, which is readily dissolved in the solution of MMA and nBA, is encapsulated within the coating after copolymerization. The copolymerization of MA-heparin with MMA and nBA significantly increases the heparin content of the coating, effectively inhibiting thrombosis and rendering the coating self-healing to help maintain long-term stability. ROS-responsive PVAX and aspirin released in a temperature-dependent manner resist acute and chronic inflammation, respectively. The heparinized self-healing and inflammation-modulated polymer coating exhibits the ability to confer long-term stability and hemocompatibility to blood-contacting biomedical materials and devices. STATEMENT OF SIGNIFICANCE: Surface engineering for blood-contacting biomedical devices paves a successful way to reduce thrombotic and inflammatory complications. However, lack of effectiveness, long-term stability and practical feasibility hinders the development and clinical application of existing strategies. Here we design a heparinized self-healing and inflammation-modulated polymer coating, which possesses high heparin level and self-healing capability to maintain long-term stability. The polymer coating is practically feasible to varied substrates and demonstrated to manipulate inflammation and prevent thrombosis both in vitro and in vivo. Our work provides a new method to develop coatings for blood-contacting biomedical materials and devices with long-term stability and hemocompatibility.

Myricetin Oligomer Triggers Multi-Receptor Mediated Penetration and Autophagic Restoration of Blood-Brain Barrier for Ischemic Stroke Treatment.

Restoration of blood-brain barrier (BBB) dysfunction, which drives worse outcomes of ischemic stroke, is a potential target for therapeutic opportunities, whereas a sealed BBB blocks the therapeutics entrance into the brain, making the BBB protection strategy paradoxical. Post ischemic stroke, hypoxia/hypoglycemia provokes the up-regulation of transmembrane glucose transporters and iron transporters due to multiple metabolic disorders, especially in brain endothelial cells. Herein, we develop a myricetin oligomer-derived nanostructure doped with Ce to bypass the BBB which is cointermediated by glucose transporters and iron transporters such as glucose transporters 1 (GLUT1), sodium/glucose cotransporters 1 (SGLT1), and transferrin(Tf) reporter (TfR). Moreover, it exhibits BBB restoration capacity by regulating the expression of tight junctions (TJs) through the activation of protective autophagy. The myricetin oligomers scaffold not only acts as targeting moiety but is the prominent active entity that inherits all diverse pharmacological activities of myricetin. The suppression of oxidative damage, M1 microglia activation, and inflammatory factors makes it a multitasking nanoagent with a single component as the scaffold, targeting domain and curative components.

Gelatin/heparin coated bio-inspired polyurethane composite fibers to construct small-caliber artificial blood vessel grafts.

Long-term patency and ability for revascularization remain challenges for small-caliber blood vessel grafts to treat cardiovascular diseases clinically. Here, a gelatin/heparin coated bio-inspired polyurethane composite fibers-based artificial blood vessel with continuous release of NO and biopeptides to regulate vascular tissue repair and maintain long-term patency is fabricated. A biodegradable polyurethane elastomer that can catalyze S-nitrosothiols in the blood to release NO is synthesized (NPU). Then, the NPU core-shell structured nanofiber grafts with requisite mechanical properties and biopeptide release for inflammation manipulation are fabricated by electrospinning and lyophilization. Finally, the surface of tubular NPU nanofiber grafts is coated with heparin/gelatin and crosslinked with glutaraldehyde to obtain small-caliber artificial blood vessels (ABVs) with the ability of vascular revascularization. We demonstrate that artificial blood vessel grafts promote the growth of endothelial cells but inhibit the growth of smooth muscle cells by the continuous release of NO; vascular grafts can regulate inflammatory balance for vascular tissue remodel without excessive collagen deposition through the release of biological peptides. Vascular grafts prevent thrombus and vascular stenosis to obtain long-term patency. Hence, our work paves a new way to develop small-caliber artificial blood vessel grafts that can maintain long-term patency in vivo and remodel vascular tissue successfully.

The potential auxiliary effects of Sargassum fusiform polysaccharides on sitagliptin in the treatment of diabetes mellitus.

This work aimed to evaluate the potential positive effects of Sargassum fusiform polysaccharides (SFP) as add-on adjuncts to sitagliptin (SIT) in treating diabetes in rats. The results showed that both SIT and SIT co-administrated with SFP (SIT+SFP) could improve hyperglycemia, glucose tolerance, insulin resistance and hyperlipidemia, and SIT+SFP exhibited better effects in alleviating the levels of blood glucose, glucose tolerance, insulin resistance index, cholesterol, and low-density lipoprotein cholesterol compared to SIT administration. Intestinal flora analysis showed that SIT+SFP treatment significantly restored the beneficial composition of gut flora as compared with SIT administration, such as the increase of Lactobacillus, Romboutsia, Blautia, Bifidobacterium, Bacteroides, Ruminococcaceae_UCG_014 and Ruminococcus_1, and the decrease of Helicobacter, Escherichia-Shigella and Pseudomonas. The fecal metabolite analysis demonstrated that the fecal bile acid and short-chain fatty acid levels in the SIT+SFP group significantly increased compared to SIT treatment. Additionally, mRNA expression results confirmed that the hypoglycemic effects of SIT+SFP were better than those of SIT, which might be attributed to the regulation of blood glucose absorption, inhibition of gluconeogenesis and regulation of cholesterol metabolism. These results suggested that SFP could be used as an auxiliary substance for SIT in treating diabetes mellitus.

Advances in Off-Target Detection for CRISPR-Based Genome Editing.

The CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-based genome editing system exhibits marked potential for both gene editing and gene therapy, and its continuous improvement contributes to its great clinical potential. However, the largest hindrance to its application in clinical practice is the presence of off-target effects (OTEs). Thus, in addition to continuous optimization of the CRISPR system to reduce and eventually eliminate OTEs, further development of unbiased genome-wide detection of OTEs is key for its successful clinical application. This article summarizes detection strategies for OTEs of different CRISPR systems, to provide detailed guidance for the detection of OTEs in CRISPR-based genome editing.

Single-injection COVID-19 subunit vaccine elicits potent immune responses.

Current vaccination schedules, including COVID-19 vaccines, require multiple doses to be administered. Single injection vaccines eliciting equivalent immune response are highly desirable. Unfortunately because unconventional release kinetics are difficult to achieve it still remains a huge challenge. Herein a single-injection COVID-19 vaccine was designed using a highly programmable release system based on dynamic layer-by-layer (LBL) films. The antigen, S1 subunit of SARS-CoV-2 spike protein, was loaded in CaCO3 microspheres, which were further coated with tannic acid (TA)/polyethylene glycol (PEG) LBL films. The single-injection vaccine was obtained by mixing the microspheres coated with different thickness of TA/PEG films. Because of the unique constant-rate erosion behavior of the TA/PEG coatings, this system allows for distinct multiple pulsatile release of antigen, closely mimicking the release profile of antigen in conventional multiple dose vaccines. Immunization with the single injection vaccine induces potent and persistent S1-specific humoral and cellular immune responses in mice. The sera from the vaccinated animal exhibit robust in vitro viral neutralization ability. More importantly, the immune response and viral inhibition induced by the single injection vaccine are as strong as that induced by the corresponding multiple dose vaccine, because they share the same antigen release profile. STATEMENT OF SIGNIFICANCE: Vaccines are the most powerful and cost-effective weapons against infectious diseases such as COVID-19. However, current vaccination schedules, including the COVID-19 vaccines, require multiple doses to be administered. Herein a single-injection COVID-19 vaccine is designed using a highly programmable release system. This vaccine releases antigens in a pulsatile manner, closely mimicking the release pattern of antigens in conventional multiple dose vaccines. As a result, one single injection of the new vaccine induces an immune response and viral inhibition similar to that induced by the corresponding multiple-dose vaccine approach.

Construction of single-injection vaccine using new time-controlled release system.

Single-injection vaccines may overcome issues, such as high cost and poor patient compliance, of the multi-bolus regimes dominantly used in vaccination. However no such vaccine has been commercialized because time-controlled release, an unconventional release kinetics, is difficult to achieve. Here a new time-controlled release system using dynamic layer-by-layer (LBL) film as erodible coating was used to design single-injection vaccine. Unlike commonly used degradable polymers, dynamic LBL film disintegrates at a constant rate, thus allowing distinct pulsatile release of antigen at predetermined intervals. The release pattern of the single-injection vaccine mimics closely to that of ordinary multi-dose regimes. It elicits both humoral and cellular immune responses which are comparable to or even stronger than the corresponding multi-dose regime. In addition, it inhibits tumor growth more effectively. The new vaccine will not only improve patient compliance but also therapeutic outcome.

Strategies for Optimization of the Clustered Regularly Interspaced Short Palindromic Repeat-Based Genome Editing System for Enhanced Editing Specificity.

The clustered regularly interspaced short palindromic repeats (CRISPR) system is inarguably the most valuable gene editing tool ever discovered. Currently, three classes of CRISPR-based genome editing systems have been developed for gene editing, including CRISPR/CRISPR associate system (Cas) nucleases, base editors, and prime editors. Ever-evolving CRISPR technology plays an important role in medicine; however, the biggest obstacle to its use in clinical practice is the induction of off-target effects (OTEs) during targeted editing. Therefore, continuous improvement and optimization of the CRISPR system for reduction of OTEs is a major focus in the field of CRISPR research. This review aims to provide a comprehensive guide for optimization of the CRISPR-based genome editing system.

Distribution patterns of nitroaromatic compounds in the water, suspended particle and sediment of the river in a long-term industrial zone (China).

Nitroaromatic compounds are known to be hazardous to ecological and human health. To assess the status of nitroaromatic compounds contamination in the main rivers in the important industrial bases of the northeastern China, we collected water, suspended particulate matter (SPM) and sediment samples from 28 sites in the Daliao River watershed and analysed them for eight nitroaromatic compounds by gas chromatography. The total concentrations of eight nitrobenzenes in the water column including aqueous and SPM phases ranged from 740 to 15,828 ng L( - 1), with a mean concentration of 3,460 ng L( - 1). The total concentrations of eight nitrobenzenes in the sediment were 7.47 to 8,185.76 ng g( - 1), with a mean concentration of 921.98 ng g( - 1), and several times higher than those found from the Yellow River in China. 4-Nitrotoluene was the predominant contaminant in the water and sediment of the three rivers of the Daliao River watershed. 2,6-Dichloro-4-nitroaniline was generally dominant in the SPM. The levels of nitroaromatic compounds were different among different sites in the Daliao River watershed, mainly caused by the distribution of pollution sources. No obvious correlation was found between the total concentrations of eight nitrobenzenes concentrations and TOC or the slit-clay content of the sediments.

A highly programmable platform for sequential release of protein therapeutics.

Drug carriers capable of releasing multiple protein therapeutics in an appropriate sequence are highly desirable for the treatment of many diseases. However current systems only allow the sequential release of two or three proteins, and it is difficult to adjust the time intervals between them. Here to solve these problems a new system is designed. The proteins are first encapsulated in CaCO3 microspheres. Then the microspheres are coated with hydrogen-bonded tannic acid (TA)/polyethylene glycol (PEG) layer-by-layer films. The encapsulated protein does not release from the microsphere until the TA/PEG coating is fully disintegrated. As the TA/PEG coating is eroded at a constant rate, the lag time for protein release is proportional to the coating thickness. To achieve sequential release, one can simply coat the protein-encapsulated microspheres with different thickness TA/PEG films and then mix them. Both in vitro and in vivo tests demonstrate that the proteins can be released from the mixed samples in a sequence according to the thickness of the TA/PEG coatings. The time intervals between the protein releases can be facilely adjusted by adjusting the thickness of the TA/PEG coatings. In addition, sequential release of more than 3 proteins can be facilely achieved.

Tough, Resilient, Adhesive, and Anti-Freezing Hydrogels Cross-Linked with a Macromolecular Cross-Linker for Wearable Strain Sensors.

Ideal conductive hydrogels for flexible, wearable strain sensors should be tough, highly resilient, adhesive, and anti-freezing. However, such hydrogels are difficult to design. Herein, a multifunctional macromolecular cross-linker (MC) based on poly(hydroxyethyl-l-glutamine) was designed and used to synthesize the hydrogels. Cross-linking with the MC leads to a reduced inhomogeneity of the gel network. Therefore, the mechanical properties of the gels are significantly improved compared with the ordinary hydrogels cross-linked with the conventional cross-linker N,N-methylenebisacrylamide (BIS). The MC-cross-linked gels also exhibit high resilience. At the same time, replacing BIS with MC significantly improves the adhesive properties of the gel, which is attributed to the introduction of a large amount of adhesive groups with the MC. The gels can stick to various substrates including skin. The good tissue adhesiveness of the gel allows it to stick to skin by itself without using any straps or adhesive tapes when used as a flexible wearable strain sensor. Both large and subtle human movements were successfully monitored using the sensor. The signals are highly stable and reliable, thanks to the high resilience of the gel. The introduction of the polar groups also improved dramatically the anti-freezing properties of the gels. Even at -20 °C, the gels still remained highly flexible and stretchable, therefore allowing the gel-based sensor to work at sub-zero temperatures. The excellent toughness, resilience, tissue-adhesiveness, and anti-freezing properties of the gel make it a good choice for a flexible wearable sensor.

Precise and tunable time-controlled drug release system using layer-by-layer films as erodible coatings.

Unlike conventional drug carriers, time-controlled release systems do not release drug immediately, but start to release drug after a predetermined lag time. Coating a drug-loaded core with an erodible barrier is a valid way to defer drug release, however, the complicated erosion behavior of the erodible coatings makes it difficult to predict and tune the lag time. Herein we proposed that dynamic layer-by-layer films, using hydrogen-bonded poly(ethylene glycol)/tannic acid (PEG/TA) film as an example, are ideal erodible coatings, because their erosion mechanism is clear and simple, and they disintegrate at constant rate. As a proof, we demonstrated that the release of bovine serum albumin (BSA) from BMS spheres can be deferred by PEG/TA coating. More importantly, the lag time can be simply tuned by the thickness of the coating. By mixing bimodal mesoporous silica (BMS) spheres coated with different thickness PEG/TA films, multiple pulse release was achieved. Similar release patterns were also successfully achieved in vivo.

Reduction of thrombotic and inflammatory complications of polystyrene-block-polyisoprene-block-polystyrene (SIS) with one-step electrospinning.

Polystyrene-block-polyisoprene-block-polystyrene (SIS) has been used as biomaterials due to its soft and stable properties under physiological conditions. However, the thrombotic and inflammatory complications caused by SIS restrain its application as blood-contacting implant. To overcome this problem, the hydrophilic core-shell structured SIS-based microfiber with antioxidant encapsulation is fabricated with one-step reactive electrospinning. We demonstrate that the phase separation of SIS and acylated Pluronic F127 (F127-DA) components and crosslinking during electrospinning renders the microfiber blood compatible and stable under physiological condition; the encapsulation of 2-O-d-glucopyranosyl-l-ascorbic acid (AA-2G) in microfiber and subsequent release of AA-2G detoxifies the excess reactive oxygen species (ROS). The microfibers are nontoxic to cells and promote the fast growth and proliferation of human umbilical vein endothelial cells (HUVECs) in the presence of ROS; the thrombotic and inflammatory complications are effectively reduced with implant evaluation in vivo. Therefore, our work paves a new way to improve the biocompatibility of SIS, making it a promising candidate for blood contact materials.

Smart microneedle patches for rapid, and painless transdermal insulin delivery.

Insulin administration at mealtimes for the control of postprandial glucose is a major part of basal-bolus insulin therapy; however, painful subcutaneous (SC) injections lead to poor patient compliance. The microneedle (MN) patch, which allows painless transdermal drug delivery, is a promising substitute; however, it remains a big challenge to deliver insulin as rapidly as by SC injection. Here a novel MN patch is designed in which the MNs are coated with insulin/poly-l-glutamic acid (PGA) layer-by-layer (LBL) films at pH 3.0. This coating is pH-sensitive because the net charge of insulin turns from positive to negative when the pH increases from 3.0 to 7.4. As a result, when transferred to pH 7.4 media, e.g., when inserted into skin, the coating dissociates instantly and releases insulin rapidly. A brief epidermal application (<1 min) of the coated MNs is enough for complete film dissociation. More importantly, the coated MN patch exhibits a pharmacokinetic and a pharmacodynamic profile comparable to that of insulin administrated by SC injection, suggesting the coated MN patch can deliver insulin as rapidly as the SC injection. In addition, the patch exhibits excellent biocompatibility and storage stability. The new MN patch is expected to become a painless, convenient method for the control of postprandial glucose.

Biomimetic nano-NOS mediated local NO release for inhibiting cancer-associated platelet activation and disrupting tumor vascular barriers.

Platelets attribute to the hypercoagulation of blood and maintenance of the tumor vascular integrity, resulting in limited intratumoral perfusion of nanoparticle into solid tumors. To overcome these adversities, we herein present an antiplatelet strategy based on erythrocyte membrane-enveloped proteinic nanoparticles that biomimic nitric oxide synthase (NOS)with co-loading of l-Arginine (LA) and photosensitizer IR783 for local NO release and inhibition of the activation of tumor-associated platelets specifically, thereby enhancing vascular permeability and accumulation of the nanoparticles in tumors. A cRGD-immobolized membrane structure is constructed to actively target platelets and cancer cells respectively, through overexpressed integrin receptors such as integrin αIIbß3 and αvß3, accelerating the inhibition of platelet activation and endocytosis of nanoparticles by tumor cells. Bio-mimicking the arginine/NO pathway in vivo, synergistical delivery of LA and IR783 enables LA molecules readily oxidize to NO with O2 that is mediated by activated IR783, the resulted NO not only retards the activity of platelets to disrupt the vascular integrity of tumor but also enhances toxicity to cancer cells. In addition, NIR-controlled release localizes the NO spatiotemporally to tumor-associated platelets and prevents undesirable systemic bleeding substantially. The reduction of the hypercoagulable state is further demonstrated by the down-regulation of tissues factor (TF) expression in tumor cells. Our study describes a promising approach to combat cancer, which advances the biomimetic NOS system as the potent therapeutic forces toward clinic applications.