Reactive Oxygen Species-Responsive Nanoparticle Delivery of Small Interfering Ribonucleic Acid Targeting Olfactory Receptor 2 for Atherosclerosis Theranostics.

Atherosclerosis (AS) is a chronic inflammatory disorder characterized by arterial intimal lipid plaques. Small interfering ribonucleic acid (siRNA)-based therapies, with their ability to suppress specific genes with high targeting precision and minimal side effects, have shown great potential for AS treatment. However, targets of siRNA therapies based on macrophages for AS treatment are still limited. Olfactory receptor 2 (Olfr2), a potential target for plaque formation, was discovered recently. Herein, anti-Olfr2 siRNA (si-Olfr2) targeting macrophages was designed, and the theranostic platform encapsulating si-Olfr2 to target macrophages within atherosclerotic lesions was also developed, with the aim of downregulating Olfr2, as well as diagnosing AS through photoacoustic imaging (PAI) in the second near-infrared (NIR-II) window with high resolution. By utilization of a reactive oxygen species (ROS)-responsive nanocarrier system, the expression of Olfr2 on macrophages within atherosclerotic plaques was effectively downregulated, leading to the inhibition of NLR family pyrin domain containing 3 (NLRP3) inflammasome activation and interleukin-1 ß (IL-1ß) secretion, thereby reducing the formation of atherosclerotic plaques. As manifested by decreased Olfr2 expression, the lesions exhibited a significantly alleviated inflammatory response that led to reduced lipid deposition, macrophage apoptosis, and a noticeable decrease in the necrotic areas. This study provides a proof of concept for evaluating the theranostic nanoplatform to specifically deliver si-Olfr2 to lesional macrophages for AS diagnosis and treatment.

DNA Framework-Enabled 3D Organization of Antiarrhythmic Drugs for Radiofrequency Catheter Ablation.

Preorganizing molecular drugs within a microenvironment is crucial for the development of efficient and controllable therapeutic systems. Here, the use of tetrahedral DNA framework (TDF) is reported to preorganize antiarrhythmic drugs (herein doxorubicin, Dox) in 3D for catheter ablation, a minimally invasive treatment for fast heartbeats, aiming to address potential complications linked to collateral tissue damage and the post-ablation atrial fibrillation (AF) recurrence resulting from incomplete ablation. Dox preorganization within TDF transforms its random distribution into a confined, regular spatial arrangement governed by DNA. This, combined with the high affinity between Dox and DNA, significantly increases local Dox concentration. The exceptional capacity of TDF for cellular internalization leads to a 5.5-fold increase in intracellular Dox amount within cardiomyocytes, effectively promoting cellular apoptosis. In vivo investigations demonstrate that administering TDF-Dox reduces the recurrence rate of electrical conduction after radiofrequency catheter ablation (RFCA) to 37.5%, compared with the 77.8% recurrence rate in the free Dox-treated group. Notably, the employed Dox dosage exhibits negligible adverse effects in vivo. This study presents a promising treatment paradigm that strengthens the efficacy of catheter ablation and opens a new avenue for reconciling the paradox of ablation efficacy and collateral damage.

Collaborative Learning and Style-Adaptive Pooling Network for Perceptual Evaluation of Arbitrary Style Transfer.

Although the research of arbitrary style transfer (AST) has achieved great progress in recent years, few studies pay special attention to the perceptual evaluation of AST images that are usually influenced by complicated factors, such as structure-preserving, style similarity, and overall vision (OV). Existing methods rely on elaborately designed hand-crafted features to obtain quality factors and apply a rough pooling strategy to evaluate the final quality. However, the importance weights between the factors and the final quality will lead to unsatisfactory performances by simple quality pooling. In this article, we propose a learnable network, named collaborative learning and style-adaptive pooling network (CLSAP-Net) to better address this issue. The CLSAP-Net contains three parts, i.e., content preservation estimation network (CPE-Net), style resemblance estimation network (SRE-Net), and OV target network (OVT-Net). Specifically, CPE-Net and SRE-Net use the self-attention mechanism and a joint regression strategy to generate reliable quality factors for fusion and weighting vectors for manipulating the importance weights. Then, grounded on the observation that style type can influence human judgment of the importance of different factors, our OVT-Net utilizes a novel style-adaptive pooling strategy guiding the importance weights of factors to collaboratively learn the final quality based on the trained CPE-Net and SRE-Net parameters. In our model, the quality pooling process can be conducted in a self-adaptive manner because the weights are generated after understanding the style type. The effectiveness and robustness of the proposed CLSAP-Net are well validated by extensive experiments on the existing AST image quality assessment (IQA) databases. Our code will be released at https://github.com/Hangwei-Chen/CLSAP-Net.

The serum soluble scavenger with 5 domains levels: A novel biomarker for individuals with heart failure.

Background: We aimed to explore the relationship between the serum Soluble Scavenger with 5 Domains (SSC5D) levels and heart failure (HF). Methods and Results: We retrospectively enrolled 276 patients diagnosed with HF or normal during hospitalization in Shanghai General Hospital between September 2020 and December 2021. Previously published RNA sequencing data were re-analyzed to confirm the expression profile of SSC5D in failing and non-failing human and mouse heart tissues. Quantitative real-time polymerase chain reaction assay was used to quantify Ssc5d mRNA levels in murine heart tissue after myocardial infarction and transverse aortic constriction surgery. To understand the HF-induced secreted proteins profile, 1,755 secreted proteins were investigated using human dilated cardiomyopathy RNA-seq data, and the results indicated that SSC5D levels were significantly elevated in failing hearts compared to the non-failing. Using single-cell RNA sequencing data, we demonstrated that Ssc5d is predominantly expressed in cardiac fibroblasts. In a murine model of myocardial infarction or transverse aortic constriction, Ssc5d mRNA levels were markedly increased compared with those in the sham group. Similarly, serum SSC5D levels were considerably elevated in the HF group compared with the control group [15,789.35 (10,745.32-23,110.65) pg/mL, 95% CI (16,263.01-19,655.43) vs. 8,938.72 (6,154.97-12,778.81) pg/mL, 95% CI (9,337.50-11,142.93); p < 0.0001]. Moreover, serum SSC5D levels were positively correlated with N-terminal pro-B-type natriuretic peptide (R = 0.4, p = 7.9e-12) and inversely correlated with left ventricular ejection fraction (R = -0.46, p = 9.8e-16). Conclusion: We concluded that SSC5D was a specific response to HF. Serum SSC5D may function as a novel biomarker and therapeutic target for patients with HF.

Targeting HDAC6 attenuates nicotine-induced macrophage pyroptosis via NF-κB/NLRP3 pathway.

BACKGROUND AND AIMS: During the development of atherosclerosis, nicotine activates macrophage inflammation. However, whether nicotine induces macrophage pyroptosis and the underlying mechanisms remain unclear. This study aimed to investigate the role of histone deacetylase 6 (HDAC6) in nicotine-induced macrophage pyroptosis. METHODS: For the in vivo study, nicotine was administered to 8-week-old ApoE-/- mice fed a high-fat diet (HFD) for 12 weeks. TUNEL/CD68 and Caspase-1/CD68 staining was used to assess macrophage pyroptosis in plaque. For the in vitro study, Western blotting, lactic dehydrogenase activity (LDH), coimmunoprecipitation, chromatin immunoprecipitation and immunofluorescence were used to evaluate pyroptosis and related signaling pathway in RAW264.7 cells. RESULTS: A high-fat diet and nicotine upregulated macrophage pyroptosis in atherosclerotic lesions. Nicotine promoted pyroptosis in RAW264.7 cells, as evidenced by increased expression of cleaved Caspase1, NLRP3, IL-1ß, IL-18, and elevated LDH release. Inhibition of HDAC6 suppressed nicotine-induced pyroptosis, which is partly mediated by p65 acetylation and NLRP3 transcription. Silencing p65 or NLRP3 resulted in decreased pyroptosis in RAW264.7 cells. CONCLUSIONS: Nicotine induces macrophage pyroptosis in atherosclerosis through HDAC6/NF-κB/NLRP3 signaling pathway.

Nicotine Modulates CTSS (Cathepsin S) Synthesis and Secretion Through Regulating the Autophagy-Lysosomal Machinery in Atherosclerosis.

OBJECTIVE: Increased CTSS (cathepsin S) has been reported to play a critical role in atherosclerosis progression. Both CTSS synthesis and secretion are essential for exerting its functions. However, the underlying mechanisms contributing to CTSS synthesis and secretion in atherosclerosis remain unclear. Approach and Results: In this study, we showed that nicotine activated autophagy and upregulated CTSS expression in vascular smooth muscle cells and in atherosclerotic plaques. Western blotting and immunofluorescent staining showed that nicotine inhibited the mTORC1 (mammalian target of rapamycin complex 1) activity, promoted the nuclear translocation of TFEB (transcription factor EB), and upregulated the expression of CTSS. Chromatin immunoprecipitation-qualificative polymerase chain reaction, electrophoretic mobility shift assay, and luciferase reporter assay further demonstrated that TFEB directly bound to the CTSS promoter. mTORC1 inhibition by nicotine or rapamycin promoted lysosomal exocytosis and CTSS secretion. Live cell assays and IP-MS (immunoprecipitation-mass spectrometry) identified that the interactions involving Rab10 (Rab10, member RAS oncogene family) and mTORC1 control CTSS secretion. Nicotine promoted vascular smooth muscle cell migration by upregulating CTSS, and CTSS inhibition suppressed nicotine-induced atherosclerosis in vivo. CONCLUSIONS: We concluded that nicotine mediates CTSS synthesis and secretion through regulating the autophagy-lysosomal machinery, which offers a potential therapeutic target for atherosclerosis treatment.

The interaction between STAT3 and nAChRα1 interferes with nicotine-induced atherosclerosis via Akt/mTOR signaling cascade.

During atherosclerosis development, nicotine and its α1 nicotinic acetylcholine receptors (nAChRα1) activate atherogenic inflammation. However, the effect of signal transducer and activator of transcription 3 (STAT3)-related inflammatory pathways in nicotine-induced atherosclerosis has been poorly studied. This study investigated the transcriptional mechanism of STAT3 in nicotine/nAChRα1-induced atherosclerosis. In vivo, ApoE-/- mice were used to establish an atherosclerotic model. Plaque area and composition were assessed by oil red O staining and immunohistochemistry. In vitro, vascular smooth muscle cells and macrophages were used to investigate cell migration, proliferation, inflammation and related signaling pathways by Transwell migration assay, EdU assay, immunofluorescence, western blotting, coimmunoprecipitation and chromatin immunoprecipitation. nAChRα1 knockdown significantly decreases the nicotine-induced upregulation of p-STAT3, p-Akt and p-mTOR in vitro, while nAChRα1 overexpression has the opposite effects. The inhibition of STAT3 attenuated nicotine-induced atherosclerosis, by reducing the proliferation and migration of vascular smooth muscle cells and inflammation in macrophages. Moreover, there is a direct interaction between STAT3 and nAChRα1 that modulates STAT3 nuclear translocation and its binding to the Akt promoter region upon nicotine exposure. Taken together, STAT3 and nAChRα1 blockade attenuates nicotine-induced atherosclerosis by reducing the migration and proliferation of vascular smooth muscle cells and inflammation in macrophages via the Akt/mTOR pathway.

Synthesis of Oxylipin Mimics and Their Antifungal Activity against the Citrus Postharvest Pathogens.

Nine oxylipin mimics were designed and synthesized starting from d-mannose. Their antifungal activity against three citrus postharvest pathogens was evaluated by spore germination assay. The results indicated that all the compounds significantly inhibited the growth of Penicillium digitatum, Penicillium italicum and Aspergillus niger. The compound (3Z,6Z,8S,9R,10R)-octadeca-3,6-diene-8,9,10-triol (3) exhibited excellent inhibitory effect on both Penicillium digitatum (IC50 = 34 ppm) and Penicillium italicum (IC50 = 94 ppm). Their in vivo antifungal activities against citrus postharvest blue mold were tested with fruit inoculated with the pathogen Penicillium italicum. The compound (3R,4S)-methyl 3,4-dihydroxy-5-octyltetrahydrofuran-2-carboxylate (9) demonstrated significant efficacy by reducing the disease severity to 60%. The antifungal mechanism of these oxylipin mimics was postulated in which both inhibition of pathogenic mycelium and stimuli of the host oxylipin-mediated defense response played important roles.

Evaluation of a rapid detection influenza virus a antigens kit using paired serum antibody test.

PURPOSE: To evaluate the feasibility for gold immunochromatographic assay (GICA) in rapid detection of influenza virus A infection. MATERIALS AND METHODS: Seventy-three patients were enrolled. All patients contributed nasopharyngeal secretions and paired serum samples. Nasopharyngeal secretions was used for colloidal gold immunochromatographic rapid assay for influenza A virus immediately after the collection of specimen. Paired serum samples were used for the hemagglutination inhibition assay at the Centers for Disease Control and Prevention influenza network laboratory in Beijing. RESULTS: Compare GICA test to hemagglutination inhibition (HI) assay, the Kappa value was 0.402 and the p value in the paired χ² test was higher than 0.05. Therefore, the difference was not statistically significant. The sensitivity of GICA was 50.0% and the specificity was 90.2%, and the negative predictive value was 90.2%. CONCLUSION: The sensitivity for Influenza A antigen detection by using GICA is relatively low, the specificity is relatively satisfactory.

The value of early diagnostic screens for the diagnosis of influenza A, B and Mycoplasma pneumoniae infections.

BACKGROUND: Rapid diagnosis of influenza virus and Mycoplasma pneumoniae infections is of importance for therapeutic intervention. It was the aim of the study to evaluate screening tests for influenza A (Flu A), B (Flu B), and Mycoplasma pneumoniae (M. pneumoniae) infections in young patients admitted to the hospital. METHODS: 522 children and juvenile patients were admitted to the hospital because of symptoms suspecting Flu A, B or M. pneumoniae infections. Gold Immunochromatography Assays were used to screen for Flu A and Flu B and rapid identification culturing with subsequent polymerase chain reaction (PCR) was carried out at admission for identification of M. pneumoniae infections. Diagnoses were based upon seroconversion during the clinical course. RESULTS: In the current study, 26% of 522 patients were shown to be infected with Flu A and 77% had positive screens using the Gold Immunochromatography Assays. 19% of 522 patients were infected with Flu B as diagnosed by seroconversion and 89% were detected by the screen. The rapid identification culture showed that 169 of 522 patients were M. pneumoniae positive. The positive samples based on rapid identification showed negative and poor concordance with PCR tests. The consistency test between the two screening methods mentioned above showed higher kappa value in the children group than infant group. CONCLUSIONS: The results are in agreement with literature and indicate that in juvenile patients and children the screening efficiency was limited. PCR assays may be applied prior to evaluation of antibody titres in the case of M. pneumoniae infections confirming previous results. The pronounced effect of age on the outcome has to be taken into account for evaluation of the screening methods.

Pulmonary embolism excluded acute coronary syndrome by coronarography: a retrospective analysis.

BACKGROUND: Pulmonary embolism (PE) is often mistaken as acute coronary syndromes (ACS) because of the considerable overlap in their clinical features. We evaluated the factors causing misdiagnosis of PE as ACS and factors that differentiate PE from ACS to improve the diagnosis efficacy of PE. METHODS: The medical records of 22 consecutive PE patients, between 2001 and 2010, who were initially suspected of ACS were retrieved. ACS was ruled out by coronary artery angiography before a definite diagnosis of PE was given. Twenty-two contemporary cases of ACS matched by age and sex were recruited as controls. Clinical manifestations, electrocardiograms (ECG), and biomarkers of these patients were reviewed retrospectively. The factors causing misdiagnosis of PE as ACS and factors differentiating PE from ACS were evaluated. RESULTS: We found two leading causes of misdiagnosis of PE as ACS. One is that PE can resemble ACS in several clinical aspects (symptoms and signs, ECG findings, plasma cardiac troponin I, and D-dimer). The other is the insufficient recognition of PE by clinicians. Risk factors for venous thromboembolism (VTE), especially deep venous thrombosis (DVT), together with signs of PE, such as unexplained dyspnea or hypoxemia, and right ventricular pressure overload on ECGs are valuable in differentiating the two diseases. CONCLUSIONS: Differentiation between PE and ACS is sometimes challenging. Adequate awareness of the risk factors for VTE and the signs of PE are crucial in the diagnosis of PE.

Depression, anxiety and influencing factors in patients with acute pulmonary embolism.

BACKGROUND: Psychological distress has been widely studied in many cardiovascular and pulmonary diseases, but the condition in acute pulmonary embolism (APE) is unknown. The purpose of this study was to investigate levels of depression and anxiety and their influencing factors in APE patients. METHODS: Sixty consecutive patients with APE were subjected to investigation of depression and anxiety by the Beck Depression Inventory and State-Trait Anxiety Inventory, and 60 community-based subjects were enrolled as controls. APE patients were stratified as high-risk, intermediate-risk and low-risk according to the disease severity. Scores of depression and anxiety were compared by statistical analysis using paired t tests between APE patients and controls, and by analysis of variance within the APE patients with the three risk stratification. Factors influencing depression and anxiety were evaluated. RESULTS: The mean age of the patients (38 males and 22 females) was (52 ± 12) years. APE patients displayed higher scores of depression (P = 0.04) and anxiety (P = 0.001) compared with controls. Patients in the high-risk group displayed higher scores of depression (P = 0.004) and anxiety (P = 0.001) compared with those in the intermediate- and low-risk groups. Depression scores were highly correlated with anxiety scores (r = 0.60, P < 0.001). Both depression and anxiety inversely related to risk stratification (P < 0.01), age (P < 0.05), and arterial blood oxygen pressure (PaO2) (P < 0.05). Linear regression analysis showed that PaO2 was independently inversely related to both depression (P < 0.01) and anxiety (P < 0.05); risk stratification and age were independently inversely related to anxiety (P < 0.05). CONCLUSIONS: Patients of APE suffered high levels of depression and anxiety, which were negatively influenced by PaO2, risk stratification and age.

The evaluation of colloidal gold immunochromatographic assay (GICA) for rapid diagnosis of influenza A disease.

BACKGROUND: A low cost colloidal gold immunochromatographic assay (GICA) for rapid detection of influenza A virus was developed. The assay was evaluated in this study. METHODS: Six hundred and twenty-six patients were enrolled. All patients contributed two pharyngeal swabs, one used for colloidal gold immunochromatographic rapid assay for influenza A virus immediately after the collection of specimen and one used for real-time reverse transcriptase polymerase chain reaction (RT-PCR) test or virus culture at the Centers for Disease Control and Prevention (CDC) influenza network laboratory. RESULT: In reference to viral culture, GICA influenza A test demonstrated a sensitivity of 64%, a specificity of 95% and an overall accuracy of 93%. Consistency between the GICA test and virus culture assay is moderate, with Kappa being 0.46. In reference to RT-PCR, GICA test demonstrated considerable high sensitivity (74%) and specificity (86%), with Kappa value being 0.61 and overall accuracy of 81%. There was no significant difference between GICA test and virus culture/RT-PCR on the detected positive rates of influenza A cases. CONCLUSIONS: GICA is a reliable, rapid, convenient and inexpensive test for the screening and diagnosis of influenza A disease. Given its lower cost than other rapid tests, the GICA test has the great potential in the management of influenza A disease in resource-poor countries.

Evaluation of the commercial rapid Mycoplasma pneumoniae medium method using the medium PCR and serum antibody test.

BACKGROUND: Mycoplasma pneumoniae (M. pneumoniae) is a major cause of respiratory tract infection. There is no specific and simple diagnosis test for the clinic treatment. In this study, we evaluated the effectiveness of a recently developed commercial rapid M. pneumoniae medium testing method. METHODS: There were 132 patients with acute respiratory tract infection enrolled in this study. All of the patients' throat swabs were taken in the morning after admission and determined using the rapid identification culture test for M. pneumoniae. In addition the M. pneumoniae polymerase chain reaction (PCR) test and routine bacterial and fungal microorganism culture test were used to identify the organisms following culture. In addition, serum from each patient was tested for M. pneumoniae using the passive particle agglutination test. RESULTS: The rapid identification culture procedure indicated that 41 patients were M. pneumoniae positive, only 6 patients were M. pneumoniae positive based on PCR, and all of the positive cultures had fungus contamination. There were only 26 of the 41 patients whose serum M. pneumoniae tested positive. Passive particle agglutination test shows 38.6% of the patients were M. pneumoniae positive (51 out of 132). Compared with the passive particle agglutination test, the specificity, sensitivity of rapid M. pneumoniae identification culture method are 81.48% and 50.98%. CONCLUSIONS: The rapid identification of M. pneumoniae culture test is easy to perform and provides results fast, but has poor concordance with the M. pneumoniae PCR and serum M. pneumoniae antibody test due to the high fungus contamination. This commercial rapid M. pneumoniae medium method needs improvement for clinical use.

[The low molecular weight heparin on rat pulmonary surfactant associated protein A of acute pulmonary embolism].

OBJECTIVE: To explore the effect of low molecular weight heparin (LMWH) on the changes of pulmonary surfactant associated protein A (SPA) of rats in acute pulmonary embolism. METHODS: Male SD rats were injected with medical gelfoam microspheres via jugular vein to induce PE model. Rats were randomized into three groups: control group (n = 8), embolism for 2 weeks group (n = 8) and LMWH therapy group (n = 8); The LMWH therapy group were injected Nadroparin subcutaneously immediately after operation, 0.1 ml/10 kg, once every 12 h. Saline were injected into the control group instead of gelfoam granule solution without further procedure. All the rats were sacrificed at the time of 2 weeks. Pulmonary artery pressure were detected by right heart catheterization and artery blood gas were analyzed at the time of sacrifice. Lung tissue were sliced and dyed with HE to observe the embolism of pulmonary artery. Methods of RT-PCR and western blot were used to study the changes of SPA mRNA and SPA protein in lung tissue. RESULTS: In control group, embolism group and LMWH group, the pulmonary pressure were (14.2 +/- 4.1) mm Hg, (29.0 +/- 8.2) mm Hg, (25.50 +/- 2.74) mm Hg respectively (F3.01, P < 0.05); the artery oxygen blood pressure (PaO2) were (94.1 +/- 8.8) mm Hg, (73.4 +/- 14.3) mm Hg, (82.86 +/- 3.73) mm Hg respectively (F 1.31, P < 0.05); SPA mRNA in three groups were 1.43 +/- 0.51, 0.87 +/- 0.35, 1.07 +/- 0.20 respectively (F 2.87, P < 0.05); and SPA protein were 1.00 +/- 0.00, 0.52 +/- 0.32, 0.90 +/- 0.22 respectively (F 2.96, P < 0.05); Under microscope, lung tissue were seen congestion, edema, infiltration of inflammatory cells in embolism group, which were lessened in LMWH group. CONCLUSIONS: The lung SPA decrease significantly in acute pulmonary embolism, and LMWH can increase the SPA, which may be one of mechanisms of LMWH in treatment of pulmonary embolism.