Coupling between 2, 2', 4, 4'-tetrabromodiphenyl ether (BDE-47) debromination and methanogenesis in anaerobic soil microcosms.

Polybrominated diphenyl ethers (PBDEs) are persistent pollutants that may undergo microbial-mediated debromination in anoxic environments, where diverse anaerobic microbes such as methanogenic archaea co-exist. However, current understanding of the relations between PBDE pollution and methanogenic process is far from complete. To address this knowledge gap, a series of anaerobic soil microcosms were established. BDE-47 (2, 2', 4, 4'-tetrabromodiphenyl ether) was selected as a model pollutant, and electron donors were supplied to stimulate the activity of anaerobes. Debromination and methane production were monitored during the 12 weeks incubation, while obligate organohalide-respiring bacteria (OHRBs), methanogenic, and the total bacterial communities were examined at week 7 and 12. The results demonstrated slow debromination of BDE-47 in all microcosms, with considerable growth of Dehalococcoides and Dehalogenimonas over the incubation observed in most BDE-47 spiked treatments. In addition, the accumulation of intermediate metabolites positively correlated with the abundance of Dehalogenimonas at week 7, suggesting potential role of these OHRBs in debromination. Methanosarcinaceae were identified as the primary methanogenic archaea, and their abundance were correlated with the production of debrominated metabolites at week 7. Furthermore, it was observed for the first time that BDE-47 considerably enhanced methane production and increased the abundance of mcrA genes, highlighting the potential effects of PBDE pollution on climate change. This might be related to the inhibition of reductive N- and S-transforming microbes, as revealed by the quantitative microbial element cycling (QMEC) analysis. Overall, our findings shed light on the intricate interactions between PBDE and methanogenic processes, and contribute to a better understanding of the environmental fate and ecological implication of PBDE under anaerobic settings.

Whole-tumor histogram analysis of apparent diffusion coefficient maps with machine learning algorithms for predicting histologic grade of sinonasal squamous cell carcinoma: a preliminary study.

PURPOSE: Accurate histologic grade assessment is helpful for clinical decision making and prognostic assessment of sinonasal squamous cell carcinoma (SNSCC). This research aimed to explore whether whole-tumor histogram analysis of apparent diffusion coefficient (ADC) maps with machine learning algorithms can predict histologic grade of SNSCC. METHODS: One hundred and forty-seven patients with pathologically diagnosed SNSCC formed this retrospective study. Sixty-six patients were low-grade (grade I/II) and eighty-one patients were high-grade (grade III). Eighteen histogram features were obtained from quantitative ADC maps. Additionally, the mean ADC value and clinical features were analyzed for comparison with histogram features. Machine learning algorithms were applied to build the best diagnostic model for predicting histological grade. The receiver operating characteristic (ROC) curve was used to evaluate the performance of each model prediction, and the area under the ROC curve (AUC) were analyzed. RESULTS: The histogram model based on three features (10th Percentile, Mean, and 90th Percentile) with support vector machine (SVM) classifier demonstrated excellent diagnostic performance, with an AUC of 0.947 on the testing dataset. The AUC of the histogram model was similar to that of the mean ADC value model (0.947 vs 0.957; P = 0.7029). The poor diagnostic performance of the clinical model (AUC = 0.692) was improved by the combined model incorporating histogram features or mean ADC value (P < 0.05). CONCLUSION: ADC histogram analysis improved the projection of SNSCC histologic grade, compared with clinical model. The complex histogram model had comparable but not better performance than mean ADC value model.

Ethanol extract of propolis regulates type 2 diabetes in mice via metabolism and gut microbiota.

ETHNOPHARMACOLOGICAL RELEVANCE: Propolis is a traditional natural medicine with various activities such as antioxidant and anti-inflammatory, immunomodulatory, anti-tumour, gastroenteritis treatment and prevention, anti-microbial and parasitic, as well as glucose regulation and anti-diabetes, and is expected to be an anti-diabetic candidate with few side effects, but the mechanism of action of propolis on type 2 diabetes mellitus (T2DM) has not been fully elucidated. AIM OF THE STUDY: The purpose of this study was to investigate the mechanism of the effect of ethanol extract of propolis (EEP) on the regulation of blood glucose in T2DM mice. MATERIALS AND METHODS: We studied the possible mechanism of EEP on T2DM using an animal model of T2DM induced by a combination of a high-fat diet and intraperitoneal injection of streptozotocin (STZ). The experiment was divided into four groups, namely, the normal group (HC), model group (T2DM), EEP and metformin group (MET). Biochemical indexes and cytokines were measured, and the differences of metabolites in the serum were compared by 1H-NMR. In addition, the diversity of intestinal flora in feces was studied by 16S rDNA amplicon sequencing. RESULTS: The results showed that following treatment with EEP and MET, the weight-loss trend of mice was alleviated, and the fasting blood glucose, insulin secretion level, insulin resistance index, C peptide level and oral glucose tolerance level decreased, whereas the insulin sensitivity index increased, thereby EEP effectively alleviated the occurrence of T2DM and insulin resistance. Compared with the T2DM group, the concentrations of pro-inflammatory cytokines interleukin-1 beta (IL-1ß), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) decreased significantly in EEP and MET groups, whereas the concentrations of anti-inflammatory cytokine interleukin-10 (IL-10) increased significantly. Metabolomics results revealed that EEP and MET regulate carbohydrate metabolism and restore amino acid and lipid metabolism. Correlation analysis of intestinal flora in mouse feces showed that compared with the HC group, harmful bacteria such as Bilophila, Eubacterium_ventriosum_group, Mucispirillum and Desulfovibrio were found in the T2DM group, whereas the abundance of beneficial bacteria such as Lactobacillus was significantly reduced. Parabacteroides, Akkermansia, Leuconostoc, and Alloprevotella were abundantly present in the EEP group; however, the MET group showed an increase in the genus Parasutterella, which could regulate energy metabolism and insulin sensitivity. CONCLUSIONS: The results showed that EEP and MET reduce fasting blood glucose in T2DM mice, followed by alleviating insulin resistance, improving the inflammatory reaction of mice, regulating the metabolism of mice, and affecting the steady state of gut microbiota. However, the overall therapeutic effect of EEP is better than that of MET.

Correction: Chemo-photodynamic combined gene therapy and dual-modal cancer imaging achieved by pH-responsive alginate/chitosan multilayer-modified magnetic mesoporous silica nanocomposites.

Correction for 'Chemo-photodynamic combined gene therapy and dual-modal cancer imaging achieved by pH-responsive alginate/chitosan multilayer-modified magnetic mesoporous silica nanocomposites' by Hong Yang et al., Biomater. Sci., 2017, 5, 1001-1013, https://doi.org/10.1039/c7bm00043j.

The clinical features and image characteristics of Meniere's disease patients with endolymphatic hydrops confirmed by enhanced magnetic resonance imaging

Abstract Objectives: To analyze and summarize the clinical features and image characteristics of Meniere's Disease (MD) patients with Endolymphatic Hydrops (EH) confirmed by enhanced Magnetic Resonance Imaging (MRI). Methods: 252 MD patients with EH confirmed by MRI were enrolled. All patients met the diagnostic criteria forMD and underwent intravenous gadolinium injection. After 4 h, MR examinations were performed. The Nakashima grading standard was used to classify EH and evaluate its correlation with clinical features. Results: Different degrees of EH were shown in all MD patients, and 157 of the 252 (62.3%) patients showed significant EH, 95 of the 252 (37.7%) patients showed mild EH. Only 89 (35.3%) met the diagnostic criteria for definite MD, and the remaining 163 (64.7%) patients met the diagnostic criteria for probable MD. Compared with patients with unilateral EH, the symptoms of the first affected ear of patients with bilateral EH were more serious. The degree of EH was related to the degree of hearing loss (p< 0.05). Conclusion: MRI with intravenous gadolinium injection can provide a better assessment of EH in MD patients. The clinical features of MD patients with EH confirmed by enhanced MRI did not fully meet the existing diagnostic criteria for definite MD. Including the diagnosis of EH in the diagnostic criteria of MD can increase the diagnosis rate of MD. The degree and distribution of EH may be related to the degree of hearing loss. Level of evidence: 4.

Do CT values change over time and correlate with hearing in the non-surgical ears of otosclerosis patients?

BACKGROUND: High-resolution computed tomography (HRCT) is often used to diagnose otosclerosis. AIMS/OBJECTIVES: To investigate whether CT values change over time and correlate with hearing in the non-surgical ears of otosclerosis patients. MATERIALS AND METHODS: 32 patients with bilateral otosclerosis who had undergone unilateral stapedectomy were enrolled in the study. HRCT examination and pure tone audiometry were performed before and after the surgery. CT values of different regions of interest (ROIs) were measured manually and the data were collected for analysis. RESULTS: The CT value of anterior to the inner auditory meatus (AIAM) decreased by 88.5 HU (p < .05), and the changes in CT values in other ROIs showed no statistical difference. Regarding hearing, the value of air bone gap (ABG) increased by 3.6 dB (p = .020), and the average hearing deterioration rate of ABG was 3.1 dB/year. The CT value of the mid-point of the stapes footplate (MPSF) showed a certain correlation with the change of ABG (p < .05). CONCLUSIONS AND SIGNIFICANCE: The CT value of AIAM may change over time. The overall hearing of non-surgical ears in otosclerosis patients shows a deteriorating trend. Moreover, the changes in the CT value of MPSF may be related to the hearing, which needs further research.

Performance of 2D BLADE turbo gradient- and spin-echo diffusion-weighted imaging in the quantitative diagnosis of recurrent temporal bone cholesteatoma.

BACKGROUND: Diffusion-weighted imaging (DWI) has become an important tool for the detection of cholesteatoma. The purpose of this study was to explore the value of 2D BLADE turbo gradient- and spin-echo imaging (TGSE BLADE) DWI in the quantitative diagnosis of recurrent temporal bone cholesteatoma (CS). METHODS: From March 2018 to October 2021, 67 patients with suspected recurrence of temporal bone CS after assessment by clinical otorhinolaryngologists who had undergone previous ear surgery for CS were prospectively evaluated by magnetic resonance imaging (MRI). Two radiologist assessed images independently. Quantitative parameters such as signal intensity ratio (SIR) calculated using, as a reference, the inferior temporal cortex (SIRT) and the background noise (SIRN), apparent diffusion coefficient (ADC) value, and ADC ratio (with pons as reference) measured on TGSE BLADE sequences were assessed. Using receiver operating characteristic (ROC) curve analysis, the optimal threshold and diagnostic performance for diagnosing recurrent CS were determined. Pair-wise comparison of the ROC curves was performed using the area under the ROC curve (AUC). RESULTS: Finally, 44 patients were included in this study, including 25 CS and 19 non-cholesteatoma (NCS). Mean SIRT and mean SIRN on TGSE BLADE DWI were significantly higher for CS than NCS lesions (p < 0.001). Meanwhile, mean ADC values and mean ADC ratios on ADC maps were significantly lower in the CS group than in the NCS group (p < 0.001). According to ROC analysis, the diagnostic efficacy of quantitative parameters such as SIRT (AUC = 0.967), SIRN (AUC = 0.979), ADC value (AUC = 1.0), and ADC ratio (AUC = 0.983) was significantly better than that of qualitative DWI (AUC = 0.867; p = 0.007, 0.009, 0.011 and 0.037, respectively). CONCLUSIONS: Residual/recurrent temporal bone CS can be accurately detected using quantitative evaluation of TGSE BLADE DWI.

The risk variant rs11836367 contributes to breast cancer onset and metastasis by attenuating Wnt signaling via regulating NTN4 expression.

Most genome-wide association study (GWAS)-identified breast cancer-associated causal variants remain uncharacterized. To provide a framework of understanding GWAS-identified variants to function, we performed a comprehensive study of noncoding regulatory variants at the NTN4 locus (12q22) and NTN4 gene in breast cancer etiology. We find that rs11836367 is the more likely causal variant, disrupting enhancer activity in both enhancer reporter assays and endogenous genome editing experiments. The protective T allele of rs11837367 increases the binding of GATA3 to the distal enhancer and up-regulates NTN4 expression. In addition, we demonstrate that loss of NTN4 gene in mice leads to tumor earlier onset, progression, and metastasis. We discover that NTN4, as a tumor suppressor, can attenuate the Wnt signaling pathway by directly binding to Wnt ligands. Our findings bridge the gaps among breast cancer-associated single-nucleotide polymorphisms, transcriptional regulation of NTN4, and breast cancer biology, which provides previously unidentified insights into breast cancer prediction and prevention.

Polyethylene microplastic exposure and concurrent effect with Aeromonas hydrophila infection on zebrafish.

Microplastics are widely distributed in the environment, raising significant concerns owing to their potential negative effects on humans. Zebrafish were used in this study to assess the toxicity of microplastic exposure. Adult zebrafish were exposed to polyethylene (PE) microplastics with smooth clustered sphere shapes and diameters of 75-100 µm for 35 days. Survival rates of the zebrafish were not significantly affected, whereas growth rates were. Analyses on oxidative stress-related enzyme activities showed that glutathione (GSH), glutathione peroxidase (GSH-PX), and glutathione s-transferase (GST) production in the intestines was stimulated when exposed to low concentrations of microplastics (0.1 and 1 mg/L), while superoxide dismutase (SOD), catalase (CAT), GSH, and GSH-PX production was suppressed when exposed to 10 mg/L microplastics. Enzyme activities in the muscles were much less affected. Intestinal injuries and changes in colony structure in the intestines were observed in zebrafish following exposure to microplastics. After 35 days of exposure, concurrent exposure to microplastics and Aeromonas hydrophila did not increase zebrafish mortality compared with those challenged by bacteria alone. This study confirms that intestinal enzyme activities of zebrafish are altered by exposure to PE microplastics but mortality and bacterial infection were not significantly affected under the tested conditions.

Enantioseparation and dissipation of acephate and its highly toxic metabolite methamidophos in pakchoi by supercritical fluid chromatography tandem mass spectrometry.

Acephate is widely used in crops as racemate. However, the enantioselective dissipation of acephate enantiomers has not been investigated in pakchoi. A sensitive and effective approach was established for determining residues of acephate and its highly toxic metabolite methamidophos enantiomers by supercritical fluid chromatography tandem mass spectrometry. Baseline separations for their enantiomers were achieved by using a Chiralcel OD-H column. The optimal chromatographic conditions were obtained as follows: CO2 /ethanol (95/5) as mobile phase; flow rate, 3.0 mL/min; column temperature, 40°C. The mean recoveries (RSDs) of analytes were in the range of 77-83.1% (6.1-9.9%), 75.4-87.5% (9.3-13.2%), and 81.5-84.2% (7.1-13.4%) at three fortification levels (0.005, 0.05, and 0.5 mg/kg for each enantiomer) for interday assay (n = 18). The method was used to evaluate the enantioselective dissipation of acephate and methamidophos in pakchoi. S-acephate dissipated faster than R-acephate, while the concentration of R-methamidophos was higher than that of S-methamidophos during the entire study period. The results indicated that the R-enantiomer of acephate and methamidophos was preferentially enriched in pakchoi. The established analysis approach and the study data provided useful information for the rational use of acephate in agriculture.

Extracellular ATP promotes breast cancer chemoresistance via HIF-1α signaling.

We have previously demonstrated that extracellular adenosine 5'-triphosphate (ATP) promotes breast cancer cell chemoresistance. However, the underlying mechanism remains unclear. Using a cDNA microarray, we demonstrated that extracellular ATP can stimulate hypoxia-inducible factor (HIF) signaling. In this study, we report that hypoxia-inducible factor 1α (HIF-1α) was upregulated after ATP treatment and mediated the ATP-driven chemoresistance process. We aimed to investigate the mechanisms and identify potential clinically relevant targets that are involved. Using mass spectrometry, we found that aldolase A (ALDOA) interacts with HIF-1α and increases HIF-1α expression. We then demonstrated that STAT3-ALDOA mediates ATP-HIF-1α signaling and upregulates the HIF-1 target genes adrenomedullin (ADM) and phosphoinositide-dependent kinase-1 (PDK1). Moreover, we show that PI3K/AKT acts upstream of HIF-1α in ATP signaling and contributes to chemoresistance in breast cancer cells. In addition, HIF-1α-knockdown or treatment with direct HIF inhibitors combined with the ATP hydrolase apyrase in MDA-MB-231 cells induced enhanced drug sensitivity in nude BALB/c mice. We then used in vitro spheroid formation assays to demonstrate the significance of ATP-HIF-1α in mediating chemoresistance. Furthermore, considering that indirect HIF inhibitors are effective in clinical cancer therapy, we treated tumor-bearing BALB/c mice with STAT3 and PI3K/AKT inhibitors and found that the dual-targeting strategy sensitized breast cancer to cisplatin. Finally, using breast cancer tissue microarrays, we found that ATP-HIF-1α signaling is associated with cancer progression, poor prognosis, and resistance to chemotherapy. Taken together, we suggest that HIF-1α signaling is vital in ATP-driven chemoresistance and may serve as a potential target for breast cancer therapies.

Levosimendan Postconditioning Attenuates Cardiomyocyte Apoptosis after Myocardial Infarction.

BACKGROUND: Levosimendan preconditioning has been shown to attenuate myocardial apoptosis in animal models. However, protective effects of levosimendan postconditioning against myocardial apoptosis following myocardial infarction (MI) have not been evaluated. Therefore, we investigated the effects of levosimendan postconditioning on myocardial apoptosis in MI rat models. METHODS: In an anoxia/reoxygenation (A/R) model, H9c2 cells were pretreated with or without levosimendan postconditioning after which their apoptosis rates were assessed by flow cytometry, RT-qPCR, and western blot analyses. Then, postconditioning was performed with or without levosimendan in MI rat models. Myocardiocyte apoptosis was evaluated by echocardiography, TTC staining, TUNEL staining, immunohistochemical staining, RT-qPCR, and western blot analysis. RESULTS: Levosimendan postconditioning inhibited H9c2 cell apoptosis in A/R models by elevating Bcl-2 while suppressing Caspase-3 and Bax at both mRNA and protein levels. Moreover, it improved cardiac functions and reduced the left ventricle infarction area in MI rat models. Compared to the MI control group, cardiomyocyte apoptosis rates in the levosimendan postconditioning group were low. The reduced cardiomyocyte apoptosis rates were associated with downregulation of Bax and Caspase-3 as well as with upregulation of Bcl-2 at mRNA and protein levels. CONCLUSIONS: Levosimendan postconditioning of MI rat models protected against cardiomyocyte apoptosis, implying that it is a potential strategy for preventing cardiomyocyte apoptosis in the treatment of cardiac dysfunction following MI.

Inner ear MRI enhancement based on three dimensional-real IR sequence in patients with Meniere's disease after intravenous gadolinium injection: Comparison of different doses used and exploration of an appropriate dose.

OBJECTIVES: Three-dimensional inversion-recovery sequence with real reconstruction (3D-real IR) magnetic resonance imaging (MRI) can detect endolymphatic hydrops of the inner ear. We aimed to explore a appropriate dose for intravenous gadolinium injection. DESIGN: Observational prospective study. SETTING: Tertiary referral centre. PARTICIPANTS: We collected 90 unilateral definite Meniere's disease patients. MAIN OUTCOME MEASURES: All enrolled patients were divided into three groups randomly (patients in group A, B and C received gadolinium injection in 1/1.5/2 times doses respectively). After 4 h, inner ear MRI scans were applied. RESULTS: The signal intensities of B-affected ears and C-affected ears were significantly higher than A-affected ears (p < .05); however, no difference was found between B-affected ears and C-affected ears (p = .267). The same conditions also appeared in the three unaffected-ear groups. Moreover, the signal intensities of affected-ear in group A, B and C were significantly higher than that of the corresponding unaffected-ear groups (p < .05). Besides, the subjective visual evaluation scores of group B and C were significantly better than that of group A (p < .05). CONCLUSIONS: Intravenous injection of gadolinium in a single dose may be unbefitting for the inner ear imaging based on 3D-real IR MRI, both the applications of gadolinium in 1.5 times and double doses can have a good perilymphatic enhancement effect of inner ear. In order to minimise the use of dose for avoiding or mitigating the adverse reactions and renal damage, 1.5 times dose may be preferred in clinical practice.

Quantitative T1 mapping MRI for the assessment of extraocular muscle fibrosis in thyroid-associated ophthalmopathy.

PURPOSE: We aimed to investigate the performance of T1 mapping and its histological correlation with extraocular muscle fibrosis in thyroid-associated ophthalmopathy (TAO). METHODS: We prospectively recruited 12 cases of active TAO, 12 cases of inactive TAO, and 15 cases of control subjects. All participants underwent magnetic resonance imaging (MRI) scan with pre-/postcontrast T1 mapping and short-time inversion-recovery (STIR) sequence. The images were analyzed to obtain precontrast T1, extracellular-volume (ECV) fraction on T1 mapping, and signal-intensity ratio (SIR) on STIR for each rectus. Muscle biopsy was performed at lateral rectus to quantify-collagen volume fraction, glycosaminoglycan (GAG)-volume fraction, and extracellular space component. The relationship between MRI and histopathology was examined with Pearson correlation coefficient. RESULTS: The active TAO group was characterized with GAG accumulation, while the inactive TAO group presented with substantial fibrosis. The MRI parameters achieved acceptable interobserver and intraobserver agreement. The precontrast T1 and ECV remarkably increased in the TAO groups than the control group, and ECV positively correlated with collagen-volume fraction (r = 0.913) and extracellular-space component (r = 0.886) in the inactive TAO group. The SIR statistically increased in the active TAO group, and SIR positively correlated with GAG-volume fraction in all three groups. The performance of ECV (cutoff > 48.1%) to screen out extraocular muscle fibrosis in inactive TAO was 60.9% sensitivity and 93.3% specificity. CONCLUSIONS: The ECV parameter on T1 mapping MRI is a reliable tool to quantify extraocular muscle fibrosis, providing insights into noninvasive evaluation of pathological changes in TAO orbit. TRIAL REGISTRATION NUMBER: ChiCTR2000040394; Date of registration: 28 November 2020.

The features comparison between patients in the ICU and general wards and between patients with different outcomes: a 2020 COVID-19 study.

BACKGROUND: The novel 2019 coronavirus (COVID-19) has largely abated in China; however, sporadic or imported cases are still a concern, while in other countries, the COVID-19 pandemic persists as a major health crisis. METHODS: All patients enrolled in this study were diagnosed with COVID-19 from February 21, 2020 to April 14, 2020 in Wuhan. We retrospectively analyzed the patients admitted to the ICU (137 patients) and general wards (114 patients) of Wuhan Leishenshan Hospital in China. The population characteristics, symptoms, and laboratory examination results between the patients in the ICU and those in the general wards were compared. Furthermore, the differences between the deceased patients in the ICU and those discharged from the ICU were compared. RESULTS: There were significant differences between the two groups in terms of symptoms, including fever, shortness of breath, no presence of complications, presence of 1 complication, and presence of 3 or more complications (P<0.05). There were also significant differences between the patients in terms of the laboratory examination results including elevated urea nitrogen, creatinine, direct bilirubin, aspartate aminotransferase, total protein, albumin, creatine kinase, lactate dehydrogenase, procalcitonin, erythrocyte sedimentation rate, white blood cells, C-reactive protein, prothrombin time, activated partial thromboplastin time, fibrinogen, D-dimer, interleukin 6, interleukin 8, interleukin 10, interleukin 2 receptor, tumor necrosis factor-α, troponin I, phosphokinase isoenzyme-MB, and B-type natriuretic peptide; and decreased platelets, lymphocyte absolute value, and eosinophil absolute value (<0.05). There were 45 patients who died in ICU and 57 improved and discharged patients. There were significant differences between the two groups in the number of patients that had 1 complication and 3 or more complications (P<0.05). There were also significant differences in the laboratory examination results between the patients including elevated urea nitrogen, total bilirubin, direct bilirubin, aspartate aminotransferase, procalcitonin, white blood cells, interleukin 8, interleukin 10, phosphokinase isoenzyme-MB, and B-type natriuretic peptide; and decreased platelets and eosinophil absolute value (P<0.05). CONCLUSIONS: Our findings highlight that the identified determinants may help to improve treatment of COVID-19 patients, to predict the risk of developing severe illness and to optimizing arrangement of health resources.

Advanced Glycation End Products Induce Vascular Smooth Muscle Cell-Derived Foam Cell Formation and Transdifferentiate to a Macrophage-Like State.

BACKGROUND: Advanced glycation end products play an important role in diabetic atherosclerosis. The effects of advanced glycation end products (AGEs) on vascular smooth muscle cell- (VSMC-) derived foam cell formation and phenotypic transformation are unknown. METHODS: Serological and histological samples were obtained from diabetic amputation patients and accident amputation patients from the Affiliated Hospital of Jiangsu University. CD68/Actin Alpha 2 (ACTA2) coimmunofluorescence sections were used to quantify the number of VSMCs with macrophage-like phenotypes. Western blotting was used to detect the expression of the receptor of advanced glycation end products in vascular samples. Enzyme-linked immunosorbent assay (ELISA) was used to evaluate the level of serum Nε-carboxymethyl-lysine (CML). In vitro oil red O staining was used to examine lipid accumulation in VSMCs stimulated by CML. The expression of VSMCs and macrophage markers was measured by western blotting and quantitative real-time PCR. Furthermore, changes in VSMC migration and secretion were detected by the Transwell assay and ELISA. RESULTS: In the arterial plaque sections of diabetic patients, VSMCs transformed to a macrophage-like phenotype. The serum CML and RAGE levels in the plaques were significantly higher in the diabetes group than those in the healthy control group and were significantly related to the number of macrophage-like VSMCs. CML stimulation promoted intracellular lipid accumulation. However, CML stimulation decreased the expression of VSMC markers and increased the expression of macrophage phenotype markers. Finally, CML promoted smooth muscle cell migration and the secretion of proinflammatory-related factors. CONCLUSIONS: CML induces VSMC-derived foam cell formation, and VSMCs transdifferentiate to a macrophage-like state, which may be mediated by the activation of RAGE.

Extracellular ATP promotes breast cancer invasion and chemoresistance via SOX9 signaling.

Our previous research demonstrated that extracellular adenosine 5'-triphosphate (ATP) could promote breast cancer cell invasion. However, the impact of extracellular ATP on chemoresistance and the mechanisms behind ATP pro-invasion and pro-chemoresistance remain unclear. Here we aimed to determine the molecules or signaling pathways involved. cDNA microarray was performed to identify the differentially expressed genes before and after ATP treatment. As a result, Sex-determining region Y-box 9 (SOX9) was up-regulated after ATP treatment in breast cancer cells. In vitro invasion and migration assays demonstrated that knocking down SOX9 attenuated ATP-driven invasive capability. Mass spectrometry and co-IP revealed that SOX9 interacted with Janus kinase 1 (JAK1). Afterward, IL-6-JAK1-STAT3 signaling was demonstrated to promote SOX9 expression and invasion following ATP treatment. Notably, ATP-IL-6-SOX9 signaling was shown to stimulate chemoresistance in breast cancer cells. ChIP assays identified some potential SOX9 target genes, among which carcinoembryonic antigen-related cell adhesion molecule 5/6 (CEACAM5/6) was demonstrated to mediate ATP pro-invasive function, while ATP-binding cassette subfamily B member 1 (ABCB1) and ATP-binding cassette subfamily G member 2 (ABCG2) mediated ATP-driven chemoresistance. In addition, SOX9-knockdown and apyrase (an ATP hydrolase)-treated MDA-MB-231 cells illustrated decreased tumor growth and enhanced drug sensitivity in nude mice. In vitro spheroid formation assays also proved the significance of ATP-SOX9 in mediating chemoresistance. Moreover, molecules involved in ATP-SOX9 signaling were up-regulated in human breast carcinoma specimens and were associated with poor prognosis. Altogether, SOX9 signaling is vital in ATP-driven invasion and chemoresistance, which may serve as a potential target for breast cancer therapies.

Facile Removal of Phytochromes and Efficient Recovery of Pesticides Using Heteropore Covalent Organic Framework-Based Magnetic Nanospheres and Electrospun Films.

Nontargeted analysis of food safety requires selective removal of interference matrices and highly efficient recovery of chemical hazards. Porous materials such as covalent organic frameworks (COFs) show great promise in selective adsorption of matrix molecules via size selectivity. Considering the complexity of interference matrices, we prepared crystalline heteropore COFs whose two kinds of pores have comparable sizes to those of several common phytochromes, main interference matrices in vegetable sample analysis. By controlling the growth of COFs on the surface of Fe3O4 nanoparticles or by utilizing a facile co-electrospinning method, heteropore COF-based magnetic nanospheres or electrospun nanofiber films were prepared, respectively. Both the nanospheres and the films maintain the dual-pore structures of COFs and show good stability and excellent reusability. Via simple magnetic separation or immersion operation, respectively, they were successfully used for the complete removal of phytochromes and highly efficient recovery of 15 pesticides from the extracts of four vegetable samples, and the recoveries are in the range of 83.10-114.00 and 60.52-107.35%, respectively. Film-based immersion operation gives better sample pretreatment performance than the film-based filtration one. This work highlights the great application potentials of heteropore COFs in sample pretreatment for nontargeted analysis, thus opening up a new way to achieve high-performance sample preparation in many fields such as food safety analysis, environment monitoring, and so on.

Nε-Carboxymethyl-Lysine Negatively Regulates Foam Cell Migration via the Vav1/Rac1 Pathway.

BACKGROUND: Macrophage-derived foam cells play a central role in atherosclerosis, and their ultimate fate includes apoptosis, promotion of vascular inflammation, or migration to other tissues. Nε-Carboxymethyl-lysine (CML), the key active component of advanced glycation end products, induced foam cell formation and apoptosis. Previous studies have shown that the Vav1/Rac1 pathway affects the macrophage cytoskeleton and cell migration, but its role in the pathogenesis of diabetic atherosclerosis is unknown. METHODS AND RESULTS: In this study, we used anterior tibiofibular vascular samples from diabetic foot amputation patients and accident amputation patients, and histological and cytological tests were performed using a diabetic ApoE-/- mouse model and primary peritoneal macrophages, respectively. The results showed that the atherosclerotic plaques of diabetic foot amputation patients and diabetic ApoE-/- mice were larger than those of the control group. Inhibition of the Vav1/Rac1 pathway reduced vascular plaques and promoted the migration of macrophages to lymph nodes. Transwell and wound healing assays showed that the migratory ability of macrophage-derived foam cells was inhibited by CML. Cytoskeletal staining showed that advanced glycation end products inhibited the formation of lamellipodia in foam cells, and inhibition of the Vav1/Rac1 pathway restored the formation of lamellipodia. CONCLUSION: CML inhibits the migration of foam cells from blood vessels via the Vav1/Rac1 pathway, and this process affects the formation of lamellipodia.

Macrophage galectin-3 enhances intimal translocation of vascular calcification in diabetes mellitus.

The clinical risks and prognosis of diabetic vascular intimal calcification (VIC) and medial calcification (VMC) are different. This study aims to investigate the mechanism of VIC/VMC translocation. Anterior tibial arteries were collected from patients with diabetic foot amputation. The patients were then divided into VIC and VMC groups. There were plaques in all anterior tibial arteries, while the enrichment of galectin-3 in arterial plaques in the VIC group was significantly higher than that in the VMC group. Furthermore, a macrophage/vascular smooth muscle cell (VSMC) coculture system was constructed. VSMC-derived extracellular vesicles (EVs) was labeled with fluorescent probe. After macrophages were pretreated with recombinant galectin-3 protein, the migration of VSMC-derived EVs and VSMC-derived calcification was more pronounced. And anti-galectin-3 antibody can inhibit this process of EVs and calcification translocation. Then, lentivirus (LV)-treated bone marrow cells (BMCs) were transplanted into apolipoprotein E-deficient (ApoE-/-) mice, and a diabetic atherosclerosis mouse model was constructed. After 15 wk of high-fat diet, ApoE-/- mice transplanted with LV-shgalectin-3 BMCs exhibited medial calcification and a concentrated distribution of EVs in the media. In conclusion, upregulation of galectin-3 in macrophages promotes the migration of VSMC-derived EVs to the intima and induces diabetic vascular intimal calcification.NEW & NOTEWORTHY The clinical risk and prognosis of vascular intimal and medial calcification are different. Macrophage galectin-3 regulates the migration of vascular smooth muscle cell-derived extracellular vesicles and mediates diabetic vascular intimal/medial calcification translocation. This study may provide insights into the early intervention in diabetic vascular calcification.