Silicone Bioadhesive with Shear-Stiffening Effect: Rate-Responsive Adhesion Behavior and Wound Dressing Application.

Stimuli-responsive adhesives with on-demand adhesion capabilities are highly advantageous for facilitating wound healing. However, the triggering conditions of stimuli-responsive adhesives are cumbersome, even though some of them are detrimental to the adhesive and adjacent natural tissues. Herein, a novel stimuli-responsive adhesive called shear-stiffening adhesive (SSA) has been created by constructing a poly(diborosiloxane)-based silicone network for the first time, and SSA exhibits a rate-responsive adhesion behavior. Furthermore, we introduced bactericidal factors (PVP-I) into SSA and applied it as a wound dressing to promote the healing of infected wounds. Impressively, the wound dressing not only has excellent biocompatibility and long-term antibacterial properties but also performs well in accelerating wound healing. Therefore, this study provides a new strategy for the synthesis of intelligent adhesives with force rate response, which simplifies the triggering conditions by the force rate. Thus, SSA has great potential to be applied in wound management as an intelligent bioadhesive with on-demand adhesion performance.

Hemostatic Tranexamic Acid-Induced Fast Gelation and Mechanical Reinforcement of Polydimethylacrylamide/Carboxymethyl Chitosan Hydrogel for Hemostasis and Wound Healing.

The combinational properties with excellent mechanical properties, adhesive performance, hemostatic ability, antibacterial action, and wound healing efficacy are highly desirable for injectable hydrogels' practical applications in hemorrhage control and wound closure, but designing one single hydrogel system integrating with such properties is still difficult. Herein, a simplified yet straightforward strategy is proposed to prepare an injectable and robust poly(N,N-dimethylacrylamide) (PDMAA)/carboxymethyl chitosan (CMCS) hydrogel induced by tranexamic acid (TXA). TXA not only promotes the rapid generation of free radicals but also introduces multiple hydrogen bonds into the hydrogel network. Moreover, as a common clinical hemostatic drug, TXA itself has excellent hemostatic effects. In addition, CMCS imparts sterilization and hemostasis effects to the hydrogel, thereby promoting wound healing. Besides, the amino and carboxyl groups on TXA molecules and the hydroxyl, amino, and carboxyl groups on CMCS molecules can form multiple hydrogen bonds with wet biological tissues, leading to good wet tissue adhesion of the hydrogel. As a result, the hydrogel with excellent mechanical properties (up to 1.83 MPa at 90% compression strain), adhesion performance (up to 18.7 kPa adhesion strength to porcine skin tissue), biocompatibility, hemostatic ability, antibacterial activity, and wound healing properties can be fabricated within several minutes. These combinational advantages enable the hydrogel to efficiently stop hemorrhage (blood loss amount: 110 mg; hemostasis time: 25 s) and promote the wound healing process (wound closure rate at 2 weeks: 83%), which can be verified using rat models of liver bleeding and infected full thickness skin defect. Overall, this facile strategy to design a hydrogel incorporating such unique advantages will greatly advance the hydrogel's clinical application in rapid hemostasis and wound healing.

Transparent and Soft Crack-Resistant Bouligand Elastomers Inspired by Fish Scales.

Nature with its abundant source offers numerous inspirations for structural and engineering designs. The oriented membranes stacked with bouligand structures in the fish scales show an outstanding combination of high strength and crack resistance. Although the applications of hard biomimetic composites are reported, the structures are rarely utilized in soft materials. Inspired by the scales of various fishes, electrospun membranes are used and stacked to fabricate bouligand elastomers, including orthogonal-plywood, single-bouligand, and double-bouligand structures. The effects of different structures on the properties of elastomers are systematically investigated and possible mechanism is explained using finite element analysis (FEA). The stiffness and fatigue characteristics of these biomimetic elastomers with the above structures are improved compared with the original membranes, especially the elastomers with a single-bouligand structure, which can undergo 5 000 cycles at a maximum strain of 35% without complete failure. The crack only propagates to half of the width of the elastomer with remaining strength of 50% of its original strength. Moreover, the mechanical performance can be adjusted by regulating the proportion of the components. The excellent crack-resistant properties and transparency promote its various potential applications.

Characterization of m6A RNA methylation mediated immune heterogeneity and functional validation in hepatocellular carcinoma.

BACKGROUND: N6 -methyladenosine (m6 A) mediates RNA modification in various biological processes. It plays a key role in hepatocellular carcinoma (HCC) through regulating methyltransferase. The present study aims to analyze the correlation between the m6 A and the immune status of HCC, and to construct an m6 A-related prognostic signature for HCC. METHODS: HCC subtypes with different m6 A modification activities were identified based on the m6 A-related genes. Lasso Cox regression was applied to construct an m6 A-related prognostic model for HCC. Then, the prognostic potential of the constructed signature was evaluated and validated in the external validation dataset. Small interfering RNAs were designed to knockdown FBXO5. CCK-8 assay, Edu staining, wound healing assay, and Transwell cell invasion assay were used to detect cell proliferation, migration, and invasion ability. RESULTS: Two m6 A-related HCC subtypes were identified. The m6 A modification active group showed an immune suppressive microenvironment compared to the m6 A modification inactive group. The differentially expressed genes (DEGs) between the HCC subtypes were screened. Enrichment analysis was performed using the DEGs. Subsequently, an m6 A-related prognostic model was established. The prognostic model performed well in both training and validation datasets. Moreover, knockdown of FBXO5, one of the genes in the prognostic model, inhibited the proliferation, migration, and invasion of HepG2 cells. CONCLUSIONS: The heterogeneity of m6 A RNA methylation is associated with immune status in HCC. The constructed m6 A-related gene-based signature can predict the prognosis of HCC patients. The genes in the prognostic model also have therapeutic potential for HCC.

Ultra-Fast-Healing Glassy Hyperbranched Plastics Capable of Restoring 26.4 MPa Tensile Strength within One Minute at Room Temperature.

The growing concern regarding widespread plastic pollution has propelled the development of sustainable self-healing plastics. Although considerable efforts have been dedicated to fabricating self-healing plastics, achieving rapid healing at room temperature is extremely challenging. Herein, we have developed an ultra-fast-healing glassy polyurethane (UGPU) by designing a hyperbranched molecular structure with a high density of multiple hydrogen bonds (H-bonds) on compliant acyclic heterochains and introducing trace water to form water bridge across the fractured surfaces. The compliant acyclic heterochains allow the dense multiple hydrogen bonds to form a frozen network, enabling tensile strength of up to 70 MPa and storage modulus of 2.5 GPa. The hyperbranched structure can drive the reorganization of the H-bonding network through the high mobility of the branched chains and terminals, thereby leading to self-healing ability at room temperature. Intriguingly, the presence of trace water vapor facilitates the formation of activated layers and the rearrangement of networks across the fractured UGPU sections, thereby enabling ultra-fast self-healing at room temperature. Consequently, the restored tensile strength after healing for 1 minute achieves a historic-record of 26.4 MPa. Furthermore, the high transparency (>90%) and ultra-fast healing property of UGPU make it an excellent candidate for advanced optical and structural materials.

Adenosine signaling mediate pain transmission in the central nervous system.

Pain is a common clinical symptom that seriously affects the quality of life in a variety of patient populations. In recent years, research on the role of adenosine signaling in pain modulation has made great progress. Adenosine is a purine nucleoside and a neuromodulator, and regulates multiple physiological and pathophysiological functions through the activation of four G protein-coupled receptors, which are classified as A1, A2A, A2B, and A3 adenosine receptors (ARs). Adenosine and its receptors that are widespread in the central nervous system (CNS) play an important role in the processing of nociceptive sensory signals in different pain models. A1Rs have the highest affinity to adenosine, and the role in analgesia has been well investigated. The roles of A2ARs and A2BRs in the modulation of pain are controversial because they have both analgesic and pronociceptive effects. The analgesic effects of A3Rs are primarily manifested in neuropathic pain. In this article, we have reviewed the recent studies on ARs in the modulation of neuropathic pain, inflammatory pain, postoperative pain, and visceral pain in the CNS. Furthermore, we have outlined the pathways through which ARs contribute to pain regulation, thereby shedding light on how this mechanism can be targeted to provide effective pain relief.

Room-temperature autonomous self-healing glassy polymers with hyperbranched structure.

Glassy polymers are extremely difficult to self-heal below their glass transition temperature (T g) due to the frozen molecules. Here, we fabricate a series of randomly hyperbranched polymers (RHP) with high density of multiple hydrogen bonds, which show T g up to 49 °C and storage modulus up to 2.7 GPa. We reveal that the hyperbranched structure not only allows the external branch units and terminals of the molecules to have a high degree of mobility in the glassy state, but also leads to the coexistence of "free" and associated complementary moieties of hydrogen bonds. The free complementary moieties can exchange with the associated hydrogen bonds, enabling network reconfiguration in the glassy polymer. As a result, the RHP shows amazing instantaneous self-healing with recovered tensile strength up to 5.5 MPa within 1 min, and the self-healing efficiency increases with contacting time at room temperature without the intervention of external stimuli.

Inhibition of Src improves cardiac fibrosis in AngII-induced hypertrophy by regulating the expression of galectin-3.

BACKGROUND: The expression of Src is upregulated in the vasculature associated with cardiac hypertrophy events. Here, we aimed to explore the underlying mechanism of Src in angiotensin II (AngII)-mediated cardiac fibrosis and hypertrophy. METHODS: The heart conditional Src knockout mouse model was established and administrated with AngII. The effects of Src on the AngII-mediated cardiac hypertrophy were assessed by Hematoxylin and Eosin (HE), Masson's trichrome, immunohistochemical staining, Annexin V-FITC/PI apoptosis detection assay and Western blot analysis. RESULTS: The expression levels of galectin-3, Src and the hypertrophy marker brain natriuretic peptide (BNP), as well as the phosphorylation of Src were all elevated in heart tissues of mice with AngII-induced cardiac hypertrophy and fibrosis. Heart conditional Src knockout attenuated AngII-activated cardiac fibrosis and hypertrophy in mice. Consistently, AngII could promote the expression of Src in a dose-dependent manner and the knockout of Src impaired Ang II-mediated apoptosis and fibrosis in the cardiomyocytes. In addition, Src inhibition suppressed the expression of galectin-3 in vivo and in vitro. Specifically, AngII could upregulate the expression of galectin-3, and knockdown of galectin-3 (Gal-3) remarkably inhibited AngII-enhanced apoptosis and fibrosis in the cardiomyocytes. Furthermore, overexpression of galectin-3 reinforced Ang II-induced cell apoptosis and fibrosis that was attenuated by knockout of Src. CONCLUSIONS: Our findings indicate that Src and Gal-3 play an important role in AngII-mediated cardiac structural remodeling. Src and galectin-3 might serve as potential targets for the treatment of AngII-induced cardiac fibrosis and hypertrophy.

Web-Based Risk Prediction Tool for an Individual's Risk of HIV and Sexually Transmitted Infections Using Machine Learning Algorithms: Development and External Validation Study.

BACKGROUND: HIV and sexually transmitted infections (STIs) are major global public health concerns. Over 1 million curable STIs occur every day among people aged 15 years to 49 years worldwide. Insufficient testing or screening substantially impedes the elimination of HIV and STI transmission. OBJECTIVE: The aim of our study was to develop an HIV and STI risk prediction tool using machine learning algorithms. METHODS: We used clinic consultations that tested for HIV and STIs at the Melbourne Sexual Health Centre between March 2, 2015, and December 31, 2018, as the development data set (training and testing data set). We also used 2 external validation data sets, including data from 2019 as external "validation data 1" and data from January 2020 and January 2021 as external "validation data 2." We developed 34 machine learning models to assess the risk of acquiring HIV, syphilis, gonorrhea, and chlamydia. We created an online tool to generate an individual's risk of HIV or an STI. RESULTS: The important predictors for HIV and STI risk were gender, age, men who reported having sex with men, number of casual sexual partners, and condom use. Our machine learning-based risk prediction tool, named MySTIRisk, performed at an acceptable or excellent level on testing data sets (area under the curve [AUC] for HIV=0.78; AUC for syphilis=0.84; AUC for gonorrhea=0.78; AUC for chlamydia=0.70) and had stable performance on both external validation data from 2019 (AUC for HIV=0.79; AUC for syphilis=0.85; AUC for gonorrhea=0.81; AUC for chlamydia=0.69) and data from 2020-2021 (AUC for HIV=0.71; AUC for syphilis=0.84; AUC for gonorrhea=0.79; AUC for chlamydia=0.69). CONCLUSIONS: Our web-based risk prediction tool could accurately predict the risk of HIV and STIs for clinic attendees using simple self-reported questions. MySTIRisk could serve as an HIV and STI screening tool on clinic websites or digital health platforms to encourage individuals at risk of HIV or an STI to be tested or start HIV pre-exposure prophylaxis. The public can use this tool to assess their risk and then decide if they would attend a clinic for testing. Clinicians or public health workers can use this tool to identify high-risk individuals for further interventions.

MicroRNA-182 exacerbates blood-brain barrier (BBB) disruption by downregulating the mTOR/FOXO1 pathway in cerebral ischemia.

Cerebral ischemia causes damage to the structure and function of the blood-brain barrier (BBB) and alleviating BBB destruction will be of great significance for the treatment and prognosis of ischemic stroke. Recently, microRNAs have been shown to play a critical role in BBB integrity. However, the potential mechanism by which microRNA-182 (miR-182) affects the BBB in ischemic stroke remains unclear. We demonstrated for the first time that cerebral ischemia leads to a significant progressive increase in miR-182 after pMCAO, and bEnd.3 cells are the primary target cells of miR-182. In miR-182 KD transgenic mice, infarct volume, and BBB permeability were attenuated, and tight junction (TJ) proteins increased. Inhibition of miR-182 with an antagomir reduced OGD-induced apoptosis of bEnd.3 cells and the loss of ZO-1 and Occludin. To further explore the mechanism by which miR-182 regulates BBB integrity, we detected the apoptotic proteins Bcl-2/Bax and demonstrated that mTOR and FOXO1 were the targets of miR-182. Inhibition of mTOR/FOXO1 by rapamycin/AS1842856 decreased the ratio of Bcl-2/Bax and exacerbated TJ protein loss. Taken together, inhibition of miR-182 protects BBB integrity by reducing endothelial cell apoptosis through the mTOR/FOXO1 pathway. Thus, miR-182 may be a potential target for the treatment of BBB disruption during cerebral ischemia.

Preparation of High-Performance Composite Hydrogel Reinforced by Hydrophilic Modified Waste Rubber Powder.

In order to reduce the environmental pollution caused by waste rubber and to realize the recycling of resources, we proposed a facile method for the hydrophilic modification of waste rubber powder (HRP) and used it to reinforce a composite hydrogel. In the presence of toluene, dibenzoyl peroxide (BPO) diffused into the waste rubber powder. After the solvent was removed, BPO was adsorbed in the rubber powder, which was used to initiate the grafting polymerization of the acrylamide monomer on the rubber-water interface. As a result, the polyacrylamide (PAM) molecular chains were grafted onto the surface of the rubber powder to realize hydrophilic modification. The success of the grafting modification was confirmed by FTIR, contact angle testing, and thermogravimetric analysis. The hydrophilic modified waste rubber powder was used to reinforce the PAM hydrogel. Mechanical tests showed that the tensile strength and elongation at the break of the composite hydrogel reached 0.46 MPa and 1809%, respectively, which was much higher than those of pure PAM hydrogel. Such a phenomenon indicates that the waste rubber particles had a strengthening effect.

Leading determinants for multimorbidity in middle-aged Australian men and women: A nine-year follow-up cohort study.

Although socioeconomic, behavioural, psychological, and biological factors have been individually linked to multimorbidity, data on the importance of these factors are limited. Our study aimed to determine the leading predictors for multimorbidity of chronic conditions in middle-aged Australian adults using machine learning methods. We included 53,867 participants aged 45-64 years from the 45 and Up Study who were free of eleven predefined chronic conditions at baseline (2006-2009) in the analysis. Incident multimorbidity was defined by the co-existence of ≥2, ≥3, or ≥ 4 conditions during follow-up until December 31, 2016. The five leading predictors for multimorbidity in men were age (7.2-20.5% of total variance), body mass index (6.5-15.4%), smoking (4.0-8.3%), chicken intake (3.6-7.5%), and red meat intake (4.6-6.3%) across the three definitions. Leading predictors varied across the three definitions in women, but the four common ones were body mass index (6.3-20.1%), age (6.2-16.4%), chicken intake (4.1-8.3%), and red meat intake (4.2-4.7%). The ten leading modifiable health factors accounted for 39.4-46.1% of total variance across the three definitions. Men with 6-10 health factors had 46-54% lower risks for multimorbidity compared with those reporting ≤2. The corresponding percentage for women was 45-52%. Non-behavioural factors including psychological distress, low education and income and high relative economic disadvantage were among the leading risk factors for multimorbidity. In conclusion, modifications on behavioural factors including diets, physical activity, smoking, alcohol consumption may reduce the risk of multimorbidity in middle-aged adults, whereas individuals with low socioeconomic status or psychological distress are at the highest priority for intervention.

Upregulation of miR-219a-5p Decreases Cerebral Ischemia/Reperfusion Injury In Vitro by Targeting Pde4d.

BACKGROUND: Ischemic stroke is the leading cause of disability and death globally. Micro-RNAs (miRNAs) have been reported to play important roles in the development and pathogenesis of the nervous system. However, the exact function and mechanism of miRNAs have not been fully elucidated about brain damage caused by cerebral ischemia/reperfusion (I/R). METHODS: In this study, we explored the neuroprotective effects of miR-219a-5p on brain using an in vitro ischemia model (mouse neuroblastoma N2a cells treated with oxyglucose deprivation and reperfusion), and in vivo cerebral I/R model in mice. Western blot assay and Reverse Transcription-Polymerase Chain Reaction were used to check the expression of molecules involved. Flow cytometry and cholecystokinin were used to examine cell apoptosis, respectively. RESULTS: Our research shows that miR-219a-5p gradually decreases in cerebral I/R models in vivo and in vitro. In vitro I/R, we find that miR-219a-5p mimics provided evidently protection for cerebral I/R damage, as shown by increased cell viability and decreased the release of LDH and cell apoptosis. Mechanically, our findings indicate that miR-219a-5p binds to cAMP specific 3', 5'-cyclic phosphodiesterase 4D (PDE4D) mRNA in the 3'-UTR region, which subsequently leads to a decrease in Pde4d expression in I/R N2a cells. CONCLUSIONS: Our results provide new ideas for the study of the mechanism of cerebral ischemia/reperfusion injury, and lay the foundation for further research on the treatment of brain I/R injury. Upregulation of miR-219a-5p decreases cerebral ischemia/reperfusion injury by targeting Pde4d in vitro.

Capillary morphogenesis protein 2 is a novel prognostic biomarker and plays oncogenic roles in glioma.

Capillary morphogenesis protein 2 (CMG2) was originally identified through its participation in capillary morphogenesis, and subsequently identified as the second receptor for anthrax toxin (ANTXR2). Although tumor-associated functions of CMG2 have also been reported, the clinical significance and functional mechanism of CMG2 in glioma remain to be elucidated. We assessed the clinicopathological relevance of CMG2 in a cohort of 48 glioma patients as well as through public glioma databases, and explored the function of CMG2 using glioblastoma (GBM) models in vitro and in vivo. CMG2 overexpression was associated with increased tumor grade and poor patient survival. CMG2 promoted G2/M-phase transition during the cell cycle of GBM cells in vitro and contributed to tumor growth in vivo. We also observed that CMG2 is implicated in the activation of extracellular signal-regulated kinases (ERKs), epithelial-mesenchymal transition (EMT), migration, and invasion in GBM cells. Transcriptomic analysis of GBM cells with or without CMG2 overexpression indicated that a panel of oncogenic signaling pathways was altered with CMG2 upregulation, implying that CMG2 might orchestrate these signaling pathways to regulate the growth of GBM cells. Yes-associated protein 1 (YAP1) activity was enhanced by CMG2 overexpression but suppressed with CMG2 deficiency. Since YAP1 is critically implicated in GBM, the oncogenic roles of CMG2 in GBM cells might thus be mediated, at least partially, by YAP1. Altogether, CMG2 functioned as an oncogene in glioma cells and is a potential prognostic biomarker or therapeutic target for the clinical treatment of glioma. Copyright © 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Targeting CD146 with a 64Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas.

Given the highly heterogeneous character of brain malignancies and the associated implication for its proper diagnosis and treatment, finding biomarkers that better characterize this disease from a molecular standpoint is imperative. In this study, we evaluated CD146 as a potential molecular target for diagnosis and targeted therapy of glioblastoma multiforme (GBM), the most common and lethal brain malignancy. YY146, an anti-CD146 monoclonal antibody, was generated and radiolabeled for noninvasive positron-emission tomography (PET) imaging of orthotopic GBM models. (64)Cu-labeled YY146 preferentially accumulated in the tumors of mice bearing U87MG xenografts, which allowed the acquisition of high-contrast PET images of small tumor nodules (∼ 2 mm). Additionally, we found that tumor uptake correlated with the levels of CD146 expression in a highly specific manner. We also explored the potential therapeutic effects of YY146 on the cancer stem cell (CSC) and epithelial-to-mesenchymal (EMT) properties of U87MG cells, demonstrating that YY146 can mitigate those aggressive phenotypes. Using YY146 as the primary antibody, we performed histological studies of World Health Organization (WHO) grades I through IV primary gliomas. The positive correlation found between CD146-positive staining and high tumor grade (χ(2) = 9.028; P = 0.029) concurred with the GBM data available in The Cancer Genome Atlas (TCGA) and validated the clinical value of YY146. In addition, we demonstrate that YY146 can be used to detect CD146 in various cancer cell lines and human resected tumor tissues of multiple other tumor types (gastric, ovarian, liver, and lung), indicating a broad applicability of YY146 in solid tumors.

Establishment and application of a new HPLC qualitative and quantitative assay for Gentiana Macrophyllae Radix based on characteristic constituents of anofinic acid and its derivatives.

Gentiana Macrophylla Radix (GMR) is officially used as traditional Chinese medicine, but easily confused with Gentianae Radix et Rhizoma (GRR) and adulterants. This study aimed to establish an HPLC method for qualitative and quantitative analysis of GMR based on characteristic components, anofinic acid and its derivatives. HPLC analysis was performed on a C18 column with gradient elution using acetonitrile and 0.1% phosphoric acid as mobile phase, and detected at 240 nm by conventional methodology validation. For fingerprint analysis, RSDs of relative retention times and relative peak areas of the characteristic peaks were within 0-1.10 and 0-4.08%, respectively. For determination of 2-methoxyanofinic acid, the calibration curve showed good linearity (R2 > 0.9999) within the test range. The RSDs of precision, repeatability and stability test did not exceed 2.46, 0.83 and 1.11%, respectively. The average recoveries were between 95.08 and 103.05% with RSDs ≤2.29%. The results showed that there was no significant difference among the four species of GMR, but there were significant differences among GMR, GRR and spurious breeds by principal component analysis and hierarchical cluster analysis. Anofinic acid and its derivatives, as the characteristic markers, could be used for the identification and quality control of GMR.

Super-Resolution Fluorescence Imaging of Spatial Organization of Proteins and Lipids in Natural Rubber.

Natural rubber (NR) with proteins and lipids has superior mechanical properties to its synthetic counterpart, polyisoprene rubber. However, it is a challenge to unravel the morphology of proteins and lipids. Here we used two-color stochastic optical reconstruction microscopy (STORM) to directly visualize the spatial organization of proteins and lipids in NR. We found that the proteins and lipids form an interdispersed stabilizing layer on the surface of NR latex particles. After drying, the proteins and lipids form aggregates of up to 300 nm in diameter. The aggregates physically interact with the terminal groups of polyisoprene chains, leading to the formation of a network, which contributes to the high elasticity and mechanical property of NR. If we remove proteins in NR, the large phospholipid aggregates disintegrate into small ones. However, it does not decompose the network but rather reduces the effective cross-linking density, thus the deproteinized NR is still elastic-like with decreased mechanical property. Removing both proteins and lipids wholly decomposes the network, thus, results in a liquid-like behavior of the rubber. The STORM measurements in this paper enable more insight into the structure-property relationship of NR, which also shows a great potential of STORM in studying the fine structure of polymeric materials and nanocomposites.

Apoptosis of THP-1 Macrophages Induced by Pseudohypericin-Mediated Sonodynamic Therapy Through the Mitochondria-Caspase Pathway.

BACKGROUND/AIMS: Pseudohypericin (P-HY) and its congener hypericin are the major hydroxylated phenanthroperylenediones present in Hypericum species. Our previous study indicated that hypericin was able to function as a sonosensitizer. The potential use of P-HY as a sonosensitizer for sonodynamic therapy (SDT) requires further exploration. Thus, this study aimed to investigate the effects of P-HY-SDT on THP-1 macrophages. METHODS: THP-1 macrophages were incubated with P-HY, and cell viability was measured using a CCK-8 assay. Fluorescence microscopy assessed the intracellular reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm ) and mitochondrial permeability transition pore (mPTP) opening. Apoptotic and necrotic cell levels were measured by the flow cytometry analysis. Western blots were employed to assay BAX, Cytochrome C expression and apoptosis-related proteins. RESULTS: P-HY-SDT induced THP-1 macrophage apoptosis. The levels of ROS were significantly increased in the SDT group, resulting in both mPTP opening and ΔΨm loss, which led to apoptosis. In addition, the translocation of BAX, release of Cytochrome C and the upregulated expression of apoptosis-related proteins after P-HY-SDT were observed, all of which were reversed by N-acetyl cysteine (NAC). CONCLUSION: P-HY-SDT induced THP-1 macrophage apoptosis through the mitochondria-caspase pathway via ROS generation, the translocation of BAX and the release of Cytochrome C to regulate the mPTP opening.

Authentication of Gentiana straminea Maxim. and its substitutes based on chemical profiling of iridoids using liquid chromatography with mass spectrometry.

A liquid chromatography-electrospray ionization-ion-trap mass spectrometry (MSn ) method was established and applied for authentication of Gentiana straminea from the four substitutes, G. tibetica, G. lhassica, G. waltonii and G. robusta, based on chemical profiling of the principal iridoid glucosides aided by a quadrupole time-of-flight mass spectrometry. The fragmentation pathways of the three representative iridoid glucosides, loganic acid, gentiopicroside and sweroside, were investigated by MSn analysis in negative ion mode, which assisted the characterization of analogs detected in the chromatographic profiling of the tested Gentiana species. In total, 25 iridoids were identified or tentatively characterized from G. straminea and four substitutes, in which 7-O-(4''-O-glucosyl)coumaroyl-loganic acid and 7-O-coumaroyl-loganic acid are diagnostic in G. straminea and can serve as the proposed chemical markers to discriminate it from morphologically similar substitutes.