Deep Penetration Research Articles

Frontiers in artificial intelligence-directed light-sheet microscopy for uncovering biological phenomena and multi-organ imaging.

Light-sheet fluorescence microscopy (LSFM) introduces fast scanning of biological phenomena with deep photon penetration and minimal phototoxicity. This advancement represents a significant shift in 3-D imaging of large-scale biological tissues and 4-D (space + time) imaging of small live animals. The large data associated with LSFM requires efficient imaging acquisition and analysis with the use of artificial intelligence (AI)/machine learning (ML) algorithms. To this end, AI/ML-directed LSFM is an emerging area for multi-organ imaging and tumor diagnostics. This review will present the development of LSFM and highlight various LSFM configurations and designs for multi-scale imaging. Optical clearance techniques will be compared for effective reduction in light scattering and optimal deep-tissue imaging. This review will further depict a diverse range of research and translational applications, from small live organisms to multi-organ imaging to tumor diagnosis. In addition, this review will address AI/ML-directed imaging reconstruction, including the application of convolutional neural networks (CNNs) and generative adversarial networks (GANs). In summary, the advancements of LSFM have enabled effective and efficient post-imaging reconstruction and data analyses, underscoring LSFM's contribution to advancing fundamental and translational research.

DEVELOPMENT OF AN ALGORITHM FOR CALCULATING THE RATE CONSTANT OF HYDROCHLORIC ACID REACTION WITH A CARBONATE ROCK FORMATION

One of the main factors reducing acid stimulation efficiency is the intensive reaction of active acid in the immediate vicinity of the wellbore, which leads to insufficient completeness of reservoir rock dissolution in the remote reservoir zone. The reservoir hydrodynamic characteristics are not being improved properly, and the efficiency of oil production stimulation measures does not reach the planned values.To get high results of acid treatments it is necessary to achieve deep penetration of acid into the formation thickness. The reaction rate constant is used to compare the reactivity of different acid compositions and to select the most suitable acid composition for the geological and physical conditions of the treated object, which provides the highest treatment efficiency. It relates the initial substances concentrations with reaction products formation rate. To determine the reaction rate constant, a volumetric apparatus was created and methodological aspects were developed to work on it, allowing to minimize the measurement error and to achieve the greatest accuracy in determining the kinetic parameters of the reaction. But in the course of further investigations in the work of the existing algorithm for experimental data processing, a number of significant deficiencies distorting the results of calculations were revealed. The aim of the research is to develop a new improved algorithm for reaction rate constant calculation, which will improve the understanding of the process of determining the kinetic parameters of acid reaction with carbonate rock.To achieve this aim, the authors conducted a series of experiments according to the unified methodology, the results of which revealed ways to eliminate the identified deficiencies using a new algorithm. It includes obtaining and saving initial data, subsequent treatment and direct calculation of reaction kinetic parameters with results saving.The developed algorithm makes it possible to unify and automate the process of determining the kinetic parameters of the acid reaction with the formation rock, as well as to determine the reaction characteristics with higher accuracy and reduce the influence of the human factor on the research results, thus reducing the errors of the calculation results.

Self-expanding cellulose sponge with enhanced hemostatic ability by tannic acid/metal ion composite coating for highly effective hemostasis of difficult-to-control bleeding wounds

Refractory bleeding presents a critical, life-threatening challenge, and the goal of medical professionals and researchers has always been to achieve safe and effective hemostasis for bleeding wounds. In this study, we utilized the benefits of a self-expanding cellulose sponge to control incompressible bleeding, which is achieved this by creating a tannic acid/metal ion coating on the surface and within the pores of the sponge to improve its hemostatic effectiveness. The effects of various types and concentrations of metal ions (calcium, magnesium, iron, and zinc) on hemostatic efficiency and biosafety is systematically investigated. The results from bacteriostasis and in vitro coagulation experiments identified 0.3 wt% Fe3+ as the optimal metal ion coating. Scanning electron microscope energy spectrum analysis confirmed the uniform distribution of Fe3+ within the cellulose sponge. Furthermore, the in vivo and in vitro results demonstrated that the prepared tannic acid/Fe3+ coated composite hemostatic sponge exhibits excellent coagulation ability and biocompatibility. Both the bleeding time and theblood loss in two bleeding models are significantly reduced, showing promising potential for treating extensive surface bleeding and deep penetrating wounds. Furthermore, the straightforward preparation method for this composite hemostatic sponge facilitates additional research towards market application.

THE EFFECT OF CALCIUM HYDROXIDE PARTICLE SIZE ON DENTIN TUBULI HARDNESS

Background: Calcium hydroxide (Ca(OH)2) is an effective root canal treatment, with calcium and hydroxyl ions effectively released on day 7. However, prolonged use can diminish dentinal tubule hardness and dissolve the hydroxyapatite crystals within them. Nanoparticle Ca(OH)2 demonstrated superior antimicrobial activity compared to conventional Ca(OH)2 because of its deeper penetration into the dentinal tubules.. Purpose: This study aimed to evaluate the effect of particle size on dentinal tubule hardness. Methods: True experiment with fifteen premolars with one root canal, no caries, and apical closure were divided into three treatment groups: conventional Ca(OH)2 (group 1, n=5), Ca(OH)2 nanoparticles (group 2, n=5), and untreated (control group), n=5. All samples were incubated for 7 days, and hardness was measured using a micro-Vickers hardness tester at 1/3 of the root canal. Kruskal-Wallis test followed by Mann-whitney post hoc analysis was used to compare the mean microvickers hardness values among different groups. The level of significance was set at p<0.05. Results: The results showed that there was significant difference between conventional Ca(OH)2 (73.00 ± 2.71) and Ca(OH)2 nanoparticles (67.40 ± 0.62) p=0.01 and the control group (70.68 ± 1.70; p>0.05) and Ca(OH)2 nanoparticles p=0.03. Conclusions: The use of Ca(OH)2 nanoparticles as an intracanal medicament for 7 days reduced dentin microhardness, whereas conventional Ca(OH)2 did not result in any change in microhardness. Particle size affects the hardness of dentinal tubules.

Research on 3D Reconstruction of Paleontological Fossils Based on Photoacoustic Microscopy

Abstract The focus on three-dimensional morphological analysis of endogenous trace fossils has emerged as a prominent aspect in the study of palaeobiological fossils. Photoacoustic imaging technology, renowned for its deep penetration, high resolution and non-destructiveness, holds considerable promise in exploring fossil morphology. In this work, a solid-coupling-based photoacoustic microscopic imaging system was developed, leveraging this technology to investigate small-scale physical fossils. The proposed system enabled layered imaging of various fossil specimens, achieving a lateral resolution exceeds 35 μm and an axial resolution superior to 8.8815 μm. The three-dimensional reconstruction results were obtained through image processing algorithms. This system was proved to be efficient to imaging the small-scale physical fossils.

Mucosal Penetrative Polymeric Micelle Formulations for Insulin Delivery to the Respiratory Tract.

Effective mucosal delivery of drugs continues to pose a significant challenge owing to the formidable barrier presented by the respiratory tract mucus, which efficiently traps and clears foreign particulates. The surface characteristics of micelles dictate their ability to penetrate the respiratory tract mucus. In this study, polymeric micelles loaded with insulin (INS) were modified using mucus-penetrative polymers. We prepared and compared polyethylene glycol (PEG)-coated micelles with micelles where cell-penetrating peptide (CPP) is conjugated to PEG. Systematic investigations of the physicochemical and aerosolization properties, performance, in vitro release, mucus and cell penetration, lung function, and pharmacokinetics/pharmacodynamics (PK/PD) of polymeric micelles were performed to evaluate their interaction with the respiratory tract. The nano-micelles, with a particle size of <100 nm, exhibited a sustained-release profile. Interestingly, PEG-coated micelles exhibited higher diffusion and deeper penetration across the mucus layer. In addition, CPP-modified micelles showed enhanced in vitro cell penetration. Finally, in the PK/PD studies, the micellar solution demonstrated higher maximum concentration (Cmax) and AUC0-8h values than subcutaneously administered INS solution, along with a sustained blood glucose-lowering effect that lasted for more than 8 h. This study proposes the use of mucus-penetrating micelle formulations as prospective inhalation nano-carriers capable of efficiently transporting peptides to the respiratory tract.

Heating Up the Immune Battle: Magnetic Hyperthermia Against Cancer

Magnetic hyperthermia (MH) utilizes magnetic iron oxide nanomaterials (MIONs) to generate nano-scale heat and boost reactive oxygen species production within cells exposed to an external alternating magnetic field. Unlike conventional thermal ablation therapies that produce heat on a macro-scale, MIONs act as point source of heat inside cells, which enables MIONs-mediated MH to modulate cellular functions and fate with precision in real-time. With key benefits such as deep tissue penetration and the ability to regulate processes in a temporal-spatial and quantifiable manner, MH is now emerging as a new cancer therapy. Most intriguing is the apparent ability for MH to alter specific biological pathways associated with an anti-tumor immune response. Research efforts are now accelerating to render MH applicable to the clinical setting, with the objective of supporting the treatment of common cancers such as hepatocellular carcinoma (HCC). In this perspective paper, we highlight the recent progress made in MH, with a particular focus on its ability to manipulate anti-tumor immune mechanisms and the therapeutic advantages demonstrated thus far for HCC. We explore the current challenges in this field, and provide our perspective on the outlook for MH and its role in cancer treatment.

Time Series Forecasting for Sparse Ring-shaped Array Photoacoustic Imaging Reconstruction

Abstract Photoacoustic computed tomography (PACT), which provides high optical absorption contrast and deep acoustic penetration, plays an important role in non-invasive biomedical imaging area. As the decrease of array elements, the reconstructed image suffers from severe artifacts. Recent studies utilize deep learning methods to improve the imaging quality of PACT based on image network design, but few were reported with raw data. To address this issue, this paper proposes a Wave to Wave Convolution Gate Recurrent-Net (WWCG-Net) to reconstruct photoacoustic image based on time series acoustic signal prediction. Simulation and experiment results show the superiority of our method compared with linear interpolation (LI) and eXtreme Gradient Boosting (XGBoost) in term of suppress artifact and improve resolution.

Hadamard Coded Multiplane Wave based Ultrafast Doppler for Rat Brain Imaging

Abstract This study explores the application of Hadamard multiplane-wave coding in ultrafast power and color Doppler imaging for enhanced visualization of rat brain vasculature. By implementing an 8-order Hadamard matrix, an approximate 10 dB improvement in signal-to-noise ratio was significantly achieved, particularly in deep brain regions and the hippocampus. The enhanced imaging technique, compared to classical plane wave coherent compounding, demonstrates its potential for applications requiring deep penetration and high sensitivity to small blood flow without compromising the ultrafast frame rate.

An Acceptor-Donor-Acceptor Structured Nano-Aggregate for NIR-Triggered Interventional Photoimmunotherapy of Cervical Cancer.

Compared with conventional therapies, photoimmunotherapy offers precise targeted cancer treatment with minimal damage to healthy tissues and reduced side effects, but its efficacy may be limited by shallow light penetration and the potential for tumor resistance. Here, an acceptor-donor-acceptor (A-D-A)-structured nanoaggregate is developed with dual phototherapy, including photodynamic therapy (PDT) and photothermal therapy (PTT), triggered by single near-infrared (NIR) light. Benefiting from strong intramolecular charge transfer (ICT), the A-D-A-structured nanoaggregates exhibit broad absorption extending to the NIR region and effectively suppressed fluorescence, which enables deep penetration and efficient photothermal conversion (η=67.94%). A suitable HOMO-LUMO distribution facilitates sufficient intersystem crossing (ISC) to convert ground-state oxygen (3O2) to singlet oxygen (1O2) and superoxide anions (·O2 -), and catalyze hydroxyl radical (·OH) generation. The enhanced ICT and ISC effects endow the A-D-A structured nanoaggregates with efficient PTT and PDT for cervical cancer, inducing efficient immunogenic cell death. In combination with clinical aluminum adjuvant gel, a novel photoimmunotherapy strategy for cervical cancer is developed and demonstrated to significantly inhibit primary and metastatic tumors in orthotopic and intraperitoneal metastasis cervical cancer animal models. The noninvasive therapy strategy offers new insights for clinical early-stage and advanced cervical cancer treatment.

Elastic circuit de-constructor: a pattern to enhance resiliency in microservices

Cloud-based workloads have proliferated with the deep penetration of the internet. Microservices based handling of high volume transactions and data have become extremely popular owing to their scalability and elasticity. The major challenge that cloud-based microservice patterns face is predicting dynamic load and failure patterns, which affect resiliency and uptime. Existing Circuit breaker patterns are biased toward denying incoming requests to maintain acceptable latency values, at the cost of availability. This paper proposes the Elastic Circuit De-Constructor (ECD) pattern to address these gaps. The proposed ECD pattern addresses this challenge by dynamically adapting to changing workloads and adjusting circuit-breaking thresholds based on real-time performance metrics. The proposed ECD pattern introduces a novel ‘De-constructed’ state, that allows the ECD to identify alternate paths pre-defined by the application, ensuring user requests continue to be routed to the microservice. By leveraging ‘Availability’, ‘Latency’ and ‘Error rate’ as performance metrics, the ECD pattern is able to balance the fault tolerance and resiliency imperatives in the cloud-based microservices environment. The performance of the proposed ECD pattern has been verified against both no Circuit Breaker and a default Circuit Breaker setting.

A self-targeting MOFs nanoplatform for treating metastatic triple-negative breast cancer through tumor microenvironment remodeling and chemotherapy potentiation

Triple-negative breast cancer (TNBC) is the most aggressive and fatal subtype of breast cancer with disappointing treatment and high mortality. Tumor microenvironment (TME) plays an important role in the invasion and metastasis of TNBC through multiple complex processes. Most anti-metastatic therapies only focus on cancer cells themselves or interfering with single factors of the metastasis process, which is often related to poor outcomes. Thus, effective TNBC treatment relies on regulating multiple key metastasis-related aspects of the TME. Herein, a self-targeting Metal-Organic Frameworks (MOFs) nanoplatform (named as MTX-PEG@TPL@ZIF-8) was designed to improve treatment of TNBC through tumor microenvironment remodeling and chemotherapy potentiation. The self-targeting MOF nanoplatform is consist of ZIF-8 nanoparticles loaded triptolide (TPL) and followed by the coating with methotrexate-polyethylene glycol conjugates (MTX-PEG). Due to MTX’s affinity for the overexpressed folate receptor on tumor cell surfaces, MTX-PEG@TPL@ZIF-8 enables effective accumulation and deep penetration in the tumor area by an MTX-mediated self-targeting strategy. This MOF nanoplatform could promptly release the medication after penetrating the tumor cell, due to pH-triggered degradation. Its anti-metastasis mechanism is to inhibit tumor invasion and metastasis by down-regulating the expression of Vimentin, MMP-2 and MMP-9 and increasing the expression of E-cadherin, upregulation of cleaved caspase-3 and cleaved caspase-9 protein expression promote the apoptosis of tumor cells, thereby reducing their migration. It also downregulated the expression of VEGF and CD31 protein to inhibit the generation of neovascularization. Overall, these findings suggest the self-targeting MOF nanoplatform offers new insights into the treatment of metastatic TNBC by TME remodeling and potentiating chemotherapy.

Integrating self-hygroscopic hydrogel electrolyte and porous electrode for supercapacitor operating in variable humidity environments

The application of supercapacitors (SCs) based on the conventional polyvinyl alcohol‑potassium hydroxide (PVA-KOH) polymer electrolyte is challenged in variable humidity environments, especially in low relative humidity (RH) environments. Herein, taking advantage of the high ionic conductivity and strong hygroscopicity of highly concentrated KOH solutions, we have prepared a hygroscopic hydrogel polymer electrolyte composed of sodium alginate (SA)/PVA/poly(acrylic acid) (PAA)-KOH. The SA/PVA/PAA-KOH gel electrolyte exhibits high ionic conductivity, superior water retention and even water adsorption capability. A three-dimensional CoCu-layered double hydroxide/zeolitic imidazole framework-67 (CuCo-LDH/ZIF-67) porous electrode has also been prepared for the construction of the high-performance SC operating in variable RH environments. The CuCo-LDH/ZIF-67 porous electrode allows deep penetration of the self-hygroscopic electrolyte, facilitating electron/ion diffusion and providing fast water transport channels. As a result, our asymmetric SC show significantly improved electrochemical performance than that of SC based on conventional PVA-KOH electrolyte under different RH environments. This integration strategy can advance the practical application of SC in extreme environments.

An NSCT-Based Multifrequency GPR Data-Fusion Method for Concealed Damage Detection

Ground-penetrating radar (GPR) is widely employed as a non-destructive tool for subsurface detection of transport infrastructures. Typically, data collected by high-frequency antennas offer high resolution but limited penetration depth, whereas data from low-frequency antennas provide deeper penetration but lower resolution. To simultaneously achieve high resolution and deep penetration via a composite radargram, a Non-Subsampled Contourlet Transform (NSCT) algorithm-based multifrequency GPR data-fusion method is proposed by integrating NSCT with appropriate fusion rules, respectively, for high-frequency and low-frequency coefficients of decomposed radargrams and by incorporating quantitative assessment metrics. Despite the advantages of NSCT in image processing, its applications to GPR data fusion for concealed damage identification of transport infrastructures are rarely reported. Numerical simulation, tunnel model test, and on-site road test are conducted for performance validation. The comparison between the evaluation metrics before and after fusion demonstrates the effectiveness of the proposed fusion method. Both shallow and deep hollow targets hidden in the simulated concrete structure, real tunnel model, and road are identified through one radargram obtained by fusing different radargrams. The significance of this study is producing a high-quality composite radargram to enable multi-depth concealed damage detection and exempting human interference in the interpretation of multiple radargrams.

Microfluidic-enabled aptamer-modified liposomal probes for targeted transient triplet differential photoacoustic imaging

Photoacoustic (PA) imaging, merging high spatial resolution and deep tissue penetration, holds promise for biomedical applications, particularly cancer diagnosis. The effectiveness of PA imaging heavily relies on the use of contrast agents that enhance the PA signal. However, the efficient preparation of targeted PA contrast agents remains a challenge. Here, we report the development of microfluidic-enabled aptamer-modified liposomal probes for targeted transient triplet differential (TTD) PA imaging. Utilizing projection micro stereolithography (PµSL) 3D printing, we fabricated cost-effective microfluidic chips for the rapid synthesis of methylene blue (MB)-loaded anti-programmed cell death ligand-1 (anti-PDL1) aptamer-modified liposomes (Apt-MB-Lip). We conducted the TTD-PA imaging of Apt-MB-Lip to improve imaging sensitivity by reducing background noise. In vivo TTD-PA experiments demonstrated the superior targeting ability and sustained presence of the Apt-MB-Lip in tumors. Our findings underscore the potential of these targeted probes for advanced diagnostic and therapeutic applications.

NIR-II-Responsive Hybrid System Achieves Cascade-Augmented Antitumor Immunity via Genetic Engineering of Both Bacteria and Tumor Cells.

The combination of nanoparticles and tumor-targeting bacteria for cancer immunotherapy can overcome the shortcomings of poor nanoparticle accumulation, limited penetration, and restricted distribution. However, it remains a great challenge for the hybrid system to improve therapeutic efficacy through the simultaneous and controllable regulation of immune cells and tumor cells. Herein, a hybrid therapeutic platform is rationally designed to achieve immune cascade-augmented cancer immunotherapy. To construct the hybrids, photothermal nanoparticles responsive to light in the second near-infrared (NIR-II) region are conjugated onto the surface of engineered bacteria through pH-responsive Schiff base bonds. Taking advantage of the hypoxia targeting and deep penetration characteristics of the bacteria, the hybrids can accumulate at tumor sites. Then nanoparticles detach from the bacteria to realize genetic engineering of tumor cells, which induces tumor cell apoptosis and down-regulate the expression of programmed cell death ligand 1 to alleviate immunosuppressive tumor microenvironment. The mild photothermal heating can not only induce tumor-associated antigen release, but also trigger sustainable expression of cytokine interleukin-2. Notably, a synergistic antitumor effect is achieved between the process of p53 transfection and NIR-II light-activated genetic engineering of bacteria. This work proposes a facile strategy for the construction of hybrid system to achieve cascade-augmented cancer immunotherapy.

NIR‐II AIEgens for Infectious Diseases Phototheranostics

Pathogenic infectious diseases have persistently posed significant threats to public health. Phototheranostics, which combines the functions of diagnostic imaging and therapy, presents an extremely promising solution to block the spread of pathogens as well as the outbreak of epidemics owing to its merits of a wide‐spectrum of activity, high controllability, non‐invasiveness, and difficult to acquire resistance. Among multifarious phototheranostic agents, second near‐infrared (NIR‐II, 1000–1700 nm) aggregation‐induced emission luminogens (AIEgens) are notable by virtue of their deep penetration depth, excellent biocompatibility, balanced radiative and nonradiative decay and aggregation‐enhanced theranostic performance, making them an ideal option for combating pathogens. This minireview provides a systematical summary of the latest advancements in NIR‐II AIEgens with emphasis on the molecular design and nanoplatform formulation to fulfill high‐efficiency in treating bacterial and viral pathogens, classified by disease models. Then, the current challenges, potential opportunities, and future research directions are presented to facilitate the further progress of this emerging field.

NIR-II AIEgens for Infectious Diseases Phototheranostics.

Pathogenic infectious diseases have persistently posed significant threats to public health. Phototheranostics, which combines the functions of diagnostic imaging and therapy, presents an extremely promising solution to block the spread of pathogens as well as the outbreak of epidemics owing to its merits of a wide-spectrum of activity, high controllability, non-invasiveness, and difficult to acquire resistance. Among multifarious phototheranostic agents, second near-infrared (NIR-II, 1000-1700 nm) aggregation-induced emission luminogens (AIEgens) are notable by virtue of their deep penetration depth, excellent biocompatibility, balanced radiative and nonradiative decay and aggregation-enhanced theranostic performance, making them an ideal option for combating pathogens. This minireview provides a systematical summary of the latest advancements in NIR-II AIEgens with emphasis on the molecular design and nanoplatform formulation to fulfill high-efficiency in treating bacterial and viral pathogens, classified by disease models. Then, the current challenges, potential opportunities, and future research directions are presented to facilitate the further progress of this emerging field.

Fluorescence and photoacoustic (FL/PA) dual-modal probe: Responsive to reactive oxygen species (ROS) for atherosclerotic plaque imaging

Accurate and early detection of atherosclerosis (AS) is imperative for their effective treatment. However, fluorescence probes for efficient diagnosis of AS often encounter insufficient deep tissue penetration, which hinders the reliable assessment of plaque vulnerability. In this work, a reactive oxygen species (ROS) activated near-infrared (NIR) fluorescence and photoacoustic (FL/PA) dual model probe TPA-QO-B is developed by conjugating two chromophores (TPA-QI and O–OH) and ROS-specific group phenylboronic acid ester. The incorporation of ROS-specific group not only induces blue shift in absorbance, but also inhibits the ICT process of TPA-QO-OH, resulting an ignorable initial FL/PA signal. ROS triggers the convertion of TPA-QO-B to TPA-QO-OH, resulting in the concurrent amplification of FL/PA signal. The exceptional selectivity of TPA-QO-B towards ROS makes it effectively distinguish AS mice from the healthy. The NIR emission can achieve a tissue penetration imaging depth of 0.3 cm. Moreover, its PA775 signal possesses the capability to penetrate tissues up to a thickness of 0.8 cm, ensuring deep in vivo imaging of AS model mice in early stage. The ROS-triggered FL/PA dual signal amplification strategy improves the accuracy and addresses the deep tissue penetration problem simultaneously, providing a promising tool for in vivo tracking biomarkers in life science and preclinical applications.

Thrombin and NIR dual-responsive system driven by heat-gas for thrombus targeted therapy

Drug thrombolysis is the main means of clinical treatment for thrombotic-related diseases. However, serious bleeding complications caused by low drug delivery efficiency and secondary vascular embolism usually lead to unsatisfactory therapeutic effects. Herein, we constructed a thrombin and near-infrared (NIR) dual-responsive drug-loaded intelligent system to solve the above problems. The CREKA-modified polydopamine (PDA) nanoparticles (NPs) were crosslinked by thrombin-responsive peptide (Pep), to obtain Pep-PPC nanocarriers with special mesoporous nanostructures. Subsequently, NO donor BNN6 and thrombolytic drug UK were loaded on the surface and into the mesoporous of Pep-PPC, to get the multifunctional Pep-NO@PPC/UK. In vitro and in vivo results proved that Pep-NO@PPC/UK could effectively recognize and aggregate at the thrombus site via CREKA-mediated active targeting. Then local abundant thrombin cleaved the nanoplatform to release UK. Meanwhile, PDA converted NIR light into heat and synchronously triggered NO release. This transformed the DDS into a “heat-gas” dual propulsion nanomotor in situ, driving deep penetration of UK in thrombus tissue. Under the combined action of UK and PTT, arterial thrombosis rapidly dissolved with relatively low bleeding risk. More importantly, NO generated in situ could prevent re-embolism of blood vessels at the thrombolytic site by inhibiting activated platelet aggregation, ultimately improving vascular reperfusion rate through a combination of attack and defense mode.