Air pollution mixture associated with oxidative stress exacerbation and symptoms deterioration in allergic rhinitis patients: Evidence from a panel study.

With allergic rhinitis (AR) on the rise globally, there has been a growing focus on the role of environmental pollutants in the onset of AR. However, the potential mechanisms by how and which these pollutants exacerbate AR conditions remain unknown. This panel study of 49 patients diagnosed with AR over one year aimed to assess the individual and combined effects of short-term exposure to multiple ambient pollutants on oxidative stress, symptoms, and quality of life among patients with AR. All participants underwent four repeated assessments of health conditions and personal environmental exposures (PM2.5, O3, SO2, and NO2) over warm and cold seasons during 2017-2018. We evaluated two oxidative stress biomarkers (malondialdehyde [MDA], and superoxide dismutase [SOD]) via nasal lavage. We collected information on self-reported symptoms and quality of life using the Rhinitis Symptom Scale (SRS), the Visual Analog Scale (VAS), and the Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ) through in-person interviews. Bayesian kernel machine regression (BKMR) was used to evaluate the joint effects of pollutant mixture and identify key contributors. The results revealed a significant association of the pollutant mixture when all four pollutants were at or above their median levels, with increased oxidative stress. This was evidenced by elevated MDA and reduced SOD. We found a joint detrimental effect of the pollutant mixture on AR symptoms with a strong association with increased SRS scores, but a non-significant positive association with VAS and RQLQ scores. PM2.5, O3, and SO2 presented as the potentially primary contributors to the adverse health effects associated with the pollutant mixture in Taiyuan city. Patients with AR exposed to short-term air pollutant mixture are more likely to have greater nasal symptoms and worse quality of life from increased oxidative stress and reduced antioxidant capacity. Further research is warranted to better elucidate the underlying mechanisms.

Vitamin B6 Turnover Predicts Long-term Mortality Risk in Patients with Type 2 Diabetes.

Background: Inflammation can increase vitamin B6 uptake and catabolism. Higher vitamin B6 turnover [4-pyridoxic acid (4-PA)/pyridoxal 5'-phosphate (PLP) ratio], was associated with mortality risk in the general population. Objectives: We aimed to investigate the association between 4-PA/PLP and long-term mortality in patients with type 2 diabetes mellitus (T2DM), an inflammatory disease. Methods: In this prospective cohort study from the National Health and Nutrition Examination Survey (NHANES) cycles 2005-2010, the concentrations of 4-PA and PLP in plasma were measured using high-performance liquid chromatography, with mortality data updated to 31 December 2019. We included 2074 patients with T2DM aged between 20 and 85 y at baseline. Results: There were 739 deaths among 2279 patients with T2DM with a median follow-up of 11.83 y. In the age- and sex-adjusted COX model (model 1), 4-PA/PLP was positively associated with mortality in patients with T2DM [hazard ratio (HR) and 95% confidence interval (CI) highest compared with lowest quartiles: 35.55 (18.29, 69.09); P < 0.001], and in model 3, which was adjusted for demographics as well as inflammation, nutrition, and renal function, high 4-PA/PLP concentrations remained an independent risk factor for mortality in patients with T2DM [HR (95% CI) highest compared with lowest quartiles: 5.03 (2.46, 10.30); P < 0.001]. In restricted cubic spline (RCS), the link between 4-PA/PLP and all-cause mortality displays a positive correlation. Patients with died within the previous 2 y were excluded, the sensitivity analysis had no effect on the association between 4-PA/PLP and mortality in patients with T2DM. Finally, comparable results were found in subgroup analyses of specific-cause mortality. Conclusion: Higher vitamin B6 turnover is associated with long-term mortality risk in patients with T2DM. 4-PA/PLP may serve as a convenient prognostic marker in T2DM management.

Rapid visual nucleic acid detection of Vibrio alginolyticus by recombinase polymerase amplification combined with CRISPR/Cas13a.

Vibrio alginolyticus (V. alginolyticus) is a common pathogen in the ocean. In addition to causing serious economic losses in aquaculture, it can also infect humans. The rapid detection of nucleic acids of V. alginolyticus with high sensitivity and specificity in the field is very important for the diagnosis and treatment of infection caused by V. alginolyticus. Here, we established a simple, fast and effective molecular method for the identification of V. alginolyticus that does not rely on expensive instruments and professionals. The method integrates recombinase polymerase amplification (RPA) technology with CRISPR system in a single PCR tube. Using this method, the results can be visualized by lateral flow dipstick (LFD) in less than 50 min, we named this method RPA-CRISPR/Cas13a-LFD. The method was confirmed to achieve high specificity for the detection of V. alginolyticus with no cross-reactivity with similar Vibrio and common clinical pathogens. This diagnostic method shows high sensitivity; the detection limit of the RPA-CRISPR/Cas13a-LFD is 10 copies/µL. We successfully identified 35 V. alginolyticus strains from a total of 55 different bacterial isolates and confirmed their identity by (Matrix-assisted laser desorption ionization time-of-flight mass spectrometry, MALDI-TOF MS). We also applied this method on infected mice blood, and the results were both easily and rapidly obtained. In conclusion, RPA-CRISPR/Cas13a-LFD offers great potential as a useful tool for reliable and rapid diagnosis of V. alginolyticus infection, especially in limited conditions.

Short-term association of fine particulate matter and its constituents with oxidative stress, symptoms and quality of life in patients with allergic rhinitis: A panel study.

BACKGROUND: Short-term exposure to fine particulate matter (PM2.5) and its specific constituents might exacerbate allergic rhinitis (AR) conditions. However, the evidence is still inconclusive. METHOD: We conducted a panel study of 49 patients diagnosed with AR > 1 year prior to the study in Taiyuan, China, to investigate associations of individual exposure to PM2.5 and its constituents with oxidative parameters, symptoms, and quality of life among AR patients. All participants underwent repeated assessments of health and PM exposure at 4 time points in both the heating and nonheating seasons from June 2017 to January 2018. AR patients' oxidative parameters were assessed using nasal lavage, and their subjective symptoms and quality of life were determined through in-person interviews using a structured questionnaire. Short-term personal exposure to PM2.5 and its constituents was estimated using the time-microenvironment-activity pattern and data from the nearest air sampler, respectively. We applied mixed-effects regression models to estimate the short-term effects of PM2.5 and its constituents. RESULTS: The results showed that exposure to PM2.5 and its constituents, including BaP, PAHs, SO42-, NH4+, V, Cr, Cu, As, Se, Cd, and Pb, was significantly associated with increased oxidative stress, as indicated by an increase in the malondialdehyde (MDA) index. Exposure to PM2.5 and its components (V, Mn, Fe, Zn, As, and Se) was associated with decreased antioxidant activity, as indicated by a decrease in the superoxide dismutase (SOD) index. Additionally, increased visual analog scale (VAS) and rhinoconjunctivitis quality of life questionnaire (RQLQ) scores indicated that exposure to PM2.5 and its constituents exacerbated inflammatory symptoms and affected quality of life in AR patients. CONCLUSION: Exposure to PM2.5 and specific constituents, could exacerbate AR patients' inflammatory symptoms and adversely affect their quality of life in the heavily industrialized city of Taiyuan, China. These findings may have potential biological and policy implications.

Complex Transformer Network for Single-Angle Plane-Wave Imaging.

OBJECTIVE: Plane-wave imaging (PWI) is a high-frame-rate imaging technique that sacrifices image quality. Deep learning can potentially enhance plane-wave image quality, but processing complex in-phase and quadrature (IQ) data and suppressing incoherent signals pose challenges. To address these challenges, we present a complex transformer network (CTN) that integrates complex convolution and complex self-attention (CSA) modules. METHODS: The CTN operates in a four-step process: delaying complex IQ data from a 0° single-angle plane wave for each pixel as CTN input data; extracting reconstruction features with a complex convolution layer; suppressing irrelevant features derived from incoherent signals with two CSA modules; and forming output images with another complex convolution layer. The training labels are generated by minimum variance (MV). RESULTS: Simulation, phantom and in vivo experiments revealed that CTN produced comparable- or even higher-quality images than MV, but with much shorter computation time. Evaluation metrics included contrast ratio, contrast-to-noise ratio, generalized contrast-to-noise ratio and lateral and axial full width at half-maximum and were -11.59 dB, 1.16, 0.68, 278 µm and 329 µm for simulation, respectively, and 9.87 dB, 0.96, 0.62, 357 µm and 305 µm for the phantom experiment, respectively. In vivo experiments further indicated that CTN could significantly improve details that were previously vague or even invisible in DAS and MV images. And after being accelerated by GPU, the CTN runtime (76.03 ms) was comparable to that of delay-and-sum (DAS, 61.24 ms). CONCLUSION: The proposed CTN significantly improved the image contrast, resolution and some unclear details by the MV beamformer, making it an efficient tool for high-frame-rate imaging.

A Novel Sample-to-Answer Visual Nucleic Acid Detection System for Adenovirus Detection.

Human adenoviruses (HAdVs) are common viruses that can cause local outbreaks in schools, communities and military camps, posing a huge threat to public health. An ideal POCT device for adenovirus detection in resource-limited settings is critical to control the spread of the virus. In this study, we developed an integrated and electricity-independent sample-to-answer system that can complete nucleic acid extraction, amplification, and detection at room temperature. This system is suitable for field and on-site detection because of its rapidity, sensitivity, lack of contamination, and lack of requirements of high-precision instruments and skilled technicians. It consists of two separate modules, ALP FINA (alkaline lysis with the paper-based filtration isolation of nucleic acid) and SV RPA (sealed and visual recombinase polymerase amplification). The extraction efficiency of ALP FINA can reach 48 to 84%, which is close to that of the conventional centrifuge column. The detection sensitivity of SV RPA is close to 10 copies/µL of AdvB and AdvE without aerosol contamination after repeated operations. When SV RPA was applied to the detection of nasopharyngeal swab samples of 19 patients who were infected with AdvB or AdvE as well as 10 healthy volunteers, its sensitivity and specificity reached 100%, respectively. IMPORTANCE HAdV infections are readily transmittable and, in some instances, highly contagious. Early and rapid diagnosis is essential for disease control. In this work, we developed a portable, disposable, and modularized sample-to-answer detection system for AdvB and AdvE, which rendered the entire test to be completely independent of electricity and other laboratory infrastructure. Thus, this detection system can be applied in resource-limited settings, and it has the potential to be further developed as an early diagnosis method in the field.

The screening and expression of polysaccharide deacetylase from Caulobacter crescentus and its function analysis.

The bacterium Caulobacter crescentus secretes an adhesive polysaccharide called holdfast, which is the known strongest underwater adhesive in nature. The deacetylase encoded by hfs (holdfast synthesis) H gene is a key factor affecting the adhesion of holdfast. Its structure and function are not yet clear, and whether other polysaccharide deacetylases exist in C. crescentus is still unknown. The screening of both HfsH and its structural analogue as well as their purification from the artificial expression products of Escherichia coli is the first step to clarify these questions. Here, we determined the conserved domains of HfsH via sequence alignment among carbohydrate esterase family 4 enzymes and screened out its structural analogue (CC_2574) in C. crescentus. The recombinant HfsH and CC_2574 were effectively expressed in E. coli. Both of them were purified by chromatography from their corresponding productions in E. coli and were then functionally analyzed. The results indicated that a high deacetylase activity (61.8 U/mg) was observed in recombinant HfsH but not in CC_2574, which suggesting that HfsH might be the irreplaceable gene mediating adhesion of holdfast in C. crescentus. Moreover, the divalent metal ions Zn2+ , Mg2+ , and Mn2+ could promote the activity of recombinant HfsH at the concentration from 0.05 to 1 mM, but inhibit its activity when the concentration exceeds 1 mM. In sum, our study first realized the artificial production of polysaccharide deacetylase HfsH and its structural analogue, and further explored their functions, both of which laid the foundation for the development of new adhesive materials.

Air Pollution Health Impact Monitoring and Health Risk Assessment Technology and Its Application - China, 2006-2019.

Air pollution is a significant risk factor contributing to the burden of disease in China. Health risk assessment and management are important to reduce the impact of air pollution on public health. To help formulate standardized health risk assessment techniques, a series of studies were conducted from 2006 to 2019. Through systematic review, study of molecular mechanisms, epidemiological investigation, and health effect monitoring, the overall project established a monitoring and evaluation indicator system, a comprehensive information platform, software for automatic data cleaning, and standardized health risk assessment techniques. Technical specifications have been issued by the National Health Commission for promoting health risk assessments across China. This paper introduces the project, the research approach, its main research accomplishments, innovations, and public health significance, and describes directions for further research.

Organic Cation Diffusion-Induced Heterogeneous Viscoelasticity in Organic-Inorganic Hybrid Perovskite Polycrystalline Films.

Organic-inorganic hybrid perovskite (OIHP) polycrystalline films are the key light-absorbing layers of laminated-structure OIHP-based devices that have attracted increasing attention in photoelectronics and flexible electronics. Internal stresses induced by the mismatched responses of laminated layers to long-term and cyclic multiphysical fields generate time-dependent mechanical deformation in OIHP polycrystalline films, which makes the mechanical constitution relation of great significance. However, few studies focus on either the mechanical properties and behaviors of OIHP polycrystalline films or the underlying mechanism coupled with the grain structure and ion diffusion. Here, we uncovered the heterogeneous viscoelasticity of MAPbBr3 films strongly correlated with the grain structure. Combining experiments and modeling, we revealed that the organic cation diffusion from grain interiors to grain boundaries leads to heterogeneity in the chemical distribution and viscoelastic modulus. Our work provides the nanomechanical understanding of the OIHP polycrystalline films that are crucial for safety design and performance optimization in OIHP-based electronics.

Research Progress of Graphene Nano-Electromechanical Resonant Sensors-A Review.

Graphene nano-electromechanical resonant sensors have wide application in areas such as seawater desalination, new energy, biotechnology, and aerospace due to their small size, light weight, and high sensitivity and resolution. This review first introduces the physical and chemical properties of graphene and the research progress of four preparation processes of graphene. Next, the principle prototype of graphene resonators is analyzed, and three main methods for analyzing the vibration characteristics of a graphene resonant sheet are described: molecular structural mechanics, non-local elastic theory and molecular dynamics. Then, this paper reviews research on graphene resonator preparation, discussing the working mechanism and research status of the development of graphene resonant mass sensors, pressure sensors and inertial sensors. Finally, the difficulties in developing graphene nano-electromechanical resonant sensors are outlined and the future trend of these sensors is described.

Modeling and Analysis of a SiC Microstructure-Based Capacitive Micro-Accelerometer.

In this study, a comb-type capacitive accelerometer based on a silicon carbide (SiC) microstructure is presented and investigated by the finite element method (FEM). It has the advantages of low weight, small volume, and low cross-coupling. Compared with silicon(111) accelerometers with the same structure, it has a higher natural frequency. When the accelerometer vibrates, its resistive force consists of two main components: a viscous damping and an elastic damping force. It was found that viscous damping dominates at low frequency, and elastic damping dominates at high frequency. The second-order linear system of the accelerometer was analyzed in the time-frequency domain, and its dynamic characteristics were best when the gap between the capacitive plates was 1.23 µm. The range of this accelerometer was 0-100 g, which is 1.64 times that of a silicon(111) accelerometer with the same structure. In addition, the accelerometer could work normally at temperatures of up to 1200 °C, which is much higher than the working temperatures of silicon devices. Therefore, the proposed accelerometer showed superior performance compared to conventional silicon-based sensors for inertial measurements.

Donor-Acceptor Competition via Halide Vacancy Filling for Oxygen Detection of High Sensitivity and Stability by All-Inorganic Perovskite Films.

Oxygen detection by organic-inorganic halide perovskites (OIHPs) has demonstrated advantages in operating temperature, response time, and reversibility over traditional materials. However, OIHPs can only sense O2 in light and the unavoidable O2 exposure during detection easily induces the degradation of OIHPs. The trade-off between sensitivity and stability makes the OIHP-based oxygen sensors impractical. By replacing organic groups with Cs, the compact films of all-inorganic halide perovskites (AIHPs) that can adsorb O2 at grain boundaries in dark are developed. AIHPs show conductance increase of 1875.5% from 1 × 10-5 to 700 Torr of O2 pressure, associated with full reversibility and long-term stability. Combining experiments and modeling, this work reveals the donor-acceptor competition via halide vacancy filling leading to the modulation of carrier concentration and mobility. This work offers understandings on oxygen sensing by perovskite materials and paves the way for further optimization of AIHPs as promising oxygen sensors with high sensitivity and stability.

Effect of Mg Addition on the Microstructure and Properties of a Heat-Affected Zone in Submerged Arc Welding of an Al-Killed Low Carbon Steel.

To reveal the effect of Mg treatment on the microstructure evolution behavior in the actual steel welding process, the microstructure and properties of Al-deoxidized high-strength ship plate steel with Mg addition were analyzed after double-side submerged arc welding. It was found that the Al-Mg-O + MnS inclusion formed under 26 ppm Mg treatment could promote acicular ferrite (AF) nucleation in the coarse-grained heat-affected zone (CGHAZ) and inhibit the formation of widmanstätten ferrite and coarse grain boundary ferrite. In the fine-grained heat-affected zone (FGHAZ) and intercritical heat-affected zone (ICHAZ), polygonal ferrite and pearlite were dominant. Al-Mg-O+MnS cannot play a role in inducing AF, but the grain size of ferrite was refined by Mg addition. The impact toughness in HAZ of the Mg-added steel was higher than that of Mg-free steel. With the heat-input rising from 29.55 to 44.11 kJ/cm, it remained relatively stable in Mg-treated steel. From the fusion line to the base metal, the micro-hardness of the fusion zone, CGHAZ, ICHAZ and FGHAZ decreased to some extent after Mg addition, which means the cold cracking tendency in the welding weak zone could be reduced. Finally, the mechanisms of Mg-containing inclusion-induced AF were also systematically discussed.

H∞ State Estimation for Neural Networks With General Activation Function and Mixed Time-Varying Delays.

This article deals with H∞ state estimation of neural networks with mixed delays. In order to make full use of delay information, novel delay-product Lyapunov-Krasovskii functional (LKF) by using parameterized delay interval is first constructed. Then, generalized free-weighting-matrix integral inequality is used to estimate the derivative of LKF to reduce the conservatism. Also, a more general activation function is further applied by combining with parameterized delay interval in order to obtain a more accurate estimator model. Finally, sufficient conditions are derived to confirm that the estimation error system is asymptotically stable with a prescribed H∞ performance. Numerical examples are simulated to show the benefits of our proposed method.

FMixCutMatch for semi-supervised deep learning.

Mixed sample augmentation (MSA) has witnessed great success in the research area of semi-supervised learning (SSL) and is performed by mixing two training samples as an augmentation strategy to effectively smooth the training space. Following the insights on the efficacy of cut-mix in particular, we propose FMixCut, an MSA that combines Fourier space-based data mixing (FMix) and the proposed Fourier space-based data cutting (FCut) for labeled and unlabeled data augmentation. Specifically, for the SSL task, our approach first generates soft pseudo-labels using the model's previous predictions. The model is then trained to penalize the outputs of the FMix-generated samples so that they are consistent with their mixed soft pseudo-labels. In addition, we propose to use FCut, a new Cutout-based data augmentation strategy that adopts the two masked sample pairs from FMix for weighted cross-entropy minimization. Furthermore, by implementing two regularization techniques, namely, batch label distribution entropy maximization and sample confidence entropy minimization, we further boost the training efficiency. Finally, we introduce a dynamic labeled-unlabeled data mixing (DDM) strategy to further accelerate the convergence of the model. Combining the above process, we finally call our SSL approach as "FMixCutMatch", in short FMCmatch. As a result, the proposed FMCmatch achieves state-of-the-art performance on CIFAR-10/100, SVHN and Mini-Imagenet across a variety of SSL conditions with the CNN-13, WRN-28-2 and ResNet-18 networks. In particular, our method achieves a 4.54% test error on CIFAR-10 with 4K labels under the CNN-13 and a 41.25% Top-1 test error on Mini-Imagenet with 10K labels under the ResNet-18. Our codes for reproducing these results are publicly available at https://github.com/biuyq/FMixCutMatch.

[Phytochemical and pharmacological progress on genus Syringa].

The genus Syringa, belonging to the family Oleaceae, are distributed naturally in the European and Asian regions.This genus is composed of more than 20 species worldwide, among which about 16 species including 10 endemic ones are discovered in China.The Syringa sp.are extensively used as herbal medicine and ornamental aspects, such as the roots and stems of S. pinnatifolia, which is one of the typical Mongolian folk medicines in China for the treatment of cardiovascular and pulmonary symptoms. As a continuous research following the previous summary in 2015, the present reriew describes the phytochemical and pharmacological progress of the genus, which hopes to provide a valuable reference to its research, development and clinic application.

GEM on proliferation and apoptosis of childhood AL cells through inhibiting c-myc expression by upregulating miR-125a-3p.

Effect of gemcitabine (GEM) on proliferation and apoptosis of childhood acute leukemia (AL) cells and the mechanism of action were investigated. Bone marrow and peripheral blood of 18 newly diagnosed children with childhood AL admitted to Yidu Central Hospital of Weifang were selected, and the miR-125a-3p level in peripheral blood of healthy children and children with AL was detected by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Leukemia cells from the bone marrow of children with AL were primarily cultured and purified to observe the morphology. miR-125a-3p mimic was transfected into childhood AL cells. The cells were randomly divided into three groups: control group, GEM group and GEM + miR-125a-3p mimic group. 5-ethynyl-2'-deoxyuridine (EdU) staining assay was chosen to detect the proliferation of childhood AL cells in each group. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining assay was adopted to determine apoptosis of childhood AL cells. The protein level of c-myc was measured via western blotting. Compared with that in the healthy children, the level of miR-125a-3p in the peripheral blood of children with AL was remarkably decreased. Compared with those in the control group, GEM inhibited proliferation and promoted apoptosis of childhood AL cells, and impeded the protein expression of c-myc in these cells. Compared with those in the GEM group, GEM + miR-125a-3p mimic notably reduced the proliferation and enhanced apoptosis of cells, and the protein expression of c-myc in cells was overtly reduced. The level of miR-125a-3p in peripheral blood of children with AL is obviously decreased. It is suggested in this study that GEM can inhibit the proliferation and promote apoptosis of childhood AL cells, and the mechanism may be related to upregulated miR-125a-3p inhibiting the expression of c-myc.

PM2.5 exposure induces vascular dysfunction via NO generated by iNOS in lung of ApoE-/- mouse.

PM2.5 exposure exacerbates cardiovascular diseases via oxidative stress and inflammation, the detailed mechanism of which is unclear. In this study, the effects of oxidative stress and inflammation, as well as vascular structure and function were studied by multiple PM2.5 exposure model of ApoE-/- mice. The results indicated that NO produced by iNOS not cNOS might play important roles in inducing vascular dysfunction after PM2.5 exposure. The occurrence order and causality among NO, other oxidative stress indicators and inflammation is explored by single PM2.5 exposure. The results showed that NO generated by iNOS occurred earlier than that of other oxidative stress indicators, which was followed by the increased inflammation. Inhibition of NOS could effectively block the raise of NO, oxidative stress and inflammation after PM2.5 exposure. All in all, we firstly confirmed that NO was the initiation factor of PM2.5 exposure-induced oxidative stress, which led to inflammation and the following vascular dysfunction.

Learning Scale-Adaptive Tight Correlation Filter for Object Tracking.

In this paper, we propose a novel tracking method by formulating tracking as a correlation filtering as well as a ridge regression problem. First, we develop a tight correlation filter-based tracking framework from the signal detection perspective. In this formulation, the correlation filter is set as the same size as the target, which can make full use of the relations of the adjacent image patches and effectively exclude the influence of the background. Specifically, we point out that the novel correlation filter model can be regarded as the ridge regression model which takes into account the different importance of the samples and has the consistent objective with tracking. Second, we focus on the scale variation problem in tracking. By making use of the spatial structure of the correlation filter, the multiscale filter banks can be generated via interpolation to handle the scale estimation problem easily. Third, we present a novel distance importance-based confidence calculation model to determine the final tracking result, which not only makes use of the fine discriminability of the correlation filter but also takes the distance importance of the candidate samples into account to alleviate the impact of similar distractors. Experimental results demonstrate that our method is superior to several state-of-the-art trackers and many other correlation filter-based methods in the benchmark datasets.

Construction of a novel Staphylokinase (SAK) mutant with low immunogenicity and its evaluation in rhesus monkey.

The heterologous nature of SAK, a thrombolytic drug, elicits high titers of neutralizing antibodies, which limits its clinical use. Here, we aim to establish a SAK mutant with equivalent activity to the wild type but reduced antigenicity, which may allow for multiple injections. Biosun software was used to predict SAK antigenic epitopes, and several main epitopes were modified by gene deletion and mutation. Ten SAK mutants were constructed, and their thrombolytic activity and immunogenicity were analyzed in vitro. SAK6, with a high expression level (45%), similar thrombolytic activity, and lower antibody reaction, was chosen for in vivo analysis in rhesus monkey. In the nearly 8-month experimental period, the antibody level of the SAK6 group was significantly lower than that of the SAK group. Moreover, only 5% of SAK activity was retained, whereas 75.6% of SAK6 activity was retained after incubating with respective antiserum. Overall, these results demonstrated that SAK6, established through comprehensive site-directed mutagenesis program, had identical thrombolytic activity to SAK, low immunogenicity, and less side effects, demonstrating its efficient clinical potential for thrombus disease.