Immobilization of a broad host range phage on the peroxidase-like Fe-MOF for colorimetric determination of multiple Salmonella enterica strains in food.

A broad host range phage-based nanozyme (Fe-MOF@SalmpYZU47) was prepared for colorimetric detection of multiple Salmonella enterica strains. The isolation of a broad host range phage (SalmpYZU47) capable of infecting multiple S. enterica strains was achieved. Then, it was directly immobilized onto the Fe-MOF to prepare Fe-MOF@SalmpYZU47, exhibiting peroxidase-like activity. The peroxidase-like activity can be specifically inhibited by multiple S. enterica strains, benefiting from the broad host range capture ability of Fe-MOF@SalmpYZU47. Based on it, a colorimetric detection approach was developed for S. enterica in the range from 1.0 × 102 to 1.0 × 108 CFU mL-1, achieving a low limit of detection (LOD) of 11 CFU mL-1. The Fe-MOF@SalmpYZU47 was utilized for detecting S. enterica in authentic food samples, achieving recoveries ranging from 91.88 to 105.34%. Hence, our proposed broad host range phage-based nanozyme exhibits significant potential for application in the colorimetric detection of pathogenic bacteria.

MiR-150-5p contributes to unexplained recurrent spontaneous abortion by targeting VEGFA and downregulating the PI3K/AKT/mTOR signaling pathway.

PURPOSE: The purpose of this study is to investigate the function of miR-150-5p in URSA. METHOD: Twenty-six chorionic villous tissues were collected to examine the expression of miR-150-5p and VEGFA by using quantitative polymerase chain reaction (qPCR) and western blot assay, respectively. Transwell assay was conducted to assess the migration and invasion ability of trophoblast cells. The dual-luciferase reporter assay was applied to determine the relationship between miR-150-5p and VEGFA in vitro. Relevant signaling pathway protein expression level was measured via western blot assay. Signaling transduction inhibitor LY294002 was used to block PI3K/AKT/mTOR signaling pathway. Finally, in vivo the effect of miR-150-5p on embryonic absorption rate was evaluated in mice. RESULTS: Clinical samples revealed that miR-150-5p expression was significantly elevated in the villous tissues and serum of URSA patients. Moreover, the overexpressing of miR-150-5p could inhibit both HTR-8/SVneo cell and JAR cell migration, invasion, and restrained PI3K/AKT/mTOR signaling pathway by targeting VEGFA in vitro. This inhibitory effect of miR-150-5p could be reversed by overexpressing the gene of vascular epithelial growth factor A (VEGFA). In contrary, inhibition of miR-150-5p significantly enhanced migration, invasion ability of both HTR-8/SVneo and JAR cells, and also could stimulate PI3K/AKT/mTOR signaling pathway. This promoting effect of miR-150-5p could be ameliorated by LY294002 (PI3K inhibitor). Finally, after miR-150-5p overexpression in vivo, the embryo resorption rate in pregnant mice was increased significantly. CONCLUSIONS: Overall, these findings imply that miR-150-5p is among the key factors that regulate the pathogenesis of URSA.

Surface engineering of magnetic peroxidase mimic using bacteriophage for high-sensitivity/specificity colorimetric determination of Staphylococcus aureus in food.

Herein, we proposed surface engineering of magnetic peroxidase mimic using bacteriophage by electrostatic interaction to prepare bacteriophage SapYZU15 modified Fe3O4 (SapYZU15@Fe3O4) for colorimetric determination of S. aureus in food. SapYZU15@Fe3O4 exhibits peroxidase-like activity, catalyzing 3,3',5,5'-tetramethylbenzidine (TMB) chromogenic reaction. After introducing S. aureus, peroxidase-like activity of SapYZU15@Fe3O4 was specifically inhibited, resulting in deceleration of TMB chromogenic reaction. This phenomenon benefits from the presence of unique tail protein gene in the bacteriophage SapYZU15 genome, leading to a specific biological interaction between S. aureus and SapYZU15. On basis of this principle, SapYZU15@Fe3O4 can be employed for colorimetric determination of S. aureus with a limiting detection (LOD), calculated as low as 1.2 × 102 CFU mL-1. With this proposed method, colorimetric detection of S. aureus in food was successfully achieved. This portends that surface engineering of nanozymes using bacteriophage has great potential in the field of colorimetric detection of pathogenic bacterium in food.

Versatile dodecyl trimethyl ammonium bromide modified γ-FeOOH for simultaneous removal and determination of As(⠤).

Inorganic arsenic pollution in water spreads all over the world, tremendously threatening environmental safety and human health. Herein, versatile dodecyl trimethyl ammonium bromide modified γ-FeOOH (DTAB-γ-FeOOH) was prepared for sportive removal and visual determination of As(Ⅴ) in water. DTAB-γ-FeOOH displays a nanosheet-like structure with a high specific surface area calculated as 166.88 m2 g-1. Additionally, DTAB-γ-FeOOH shows peroxidase-mimicking feature, which can catalyze colorless TMB to generate blue oxidized TMB (TMBox) in presence of H2O2. Removal experiments show that DTAB-γ-FeOOH exhibits good As(Ⅴ) removal efficiency because modification of DTAB makes γ-FeOOH carry abundant positive charges, improving affinity between DTAB-γ-FeOOH and As(Ⅴ). It is found that theoretical maximum adsorption capacity is up to 126.91 mg g-1. Moreover, DTAB-γ-FeOOH can resist interference of most of co-existing ions. After that, As(Ⅴ) was detected based on peroxidase-like DTAB-γ-FeOOH. As(Ⅴ) can be adsorbed onto DTAB-γ-FeOOH surface, markedly inhibiting its peroxidase-like activity. Based on it, As(Ⅴ) ranging from 1.67 to 3333.33 µg L-1 can be well detected, with a low LOD (0.84 µg L-1). The successful sorptive removal and visual determination of As(Ⅴ) from real environmental water indicated that DTAB-γ-FeOOH has great potential in the treatment of As(Ⅴ)-containing environment water.

A smartphone-integrated colorimetric quantitative analysis platform based on oxidase-like Ce(IV)-ATP-Tris CPNs/CNF test strip for detection of inorganic arsenic in rice.

Herein, a colorimetric sensing system based on cerium(IV) coordination polymer nanoparticles (Ce(IV)-ATP-Tris CPNs) was proposed for As(V) detection. Ce(IV)-ATP-Tris CPNs show excellent oxidase-like activity, triggering 3,3',5,5'-tetramethylbenzidine (TMB) chromogenic reaction. With addition of ascorbic acid 2-phosphate (AAP) and acid phosphatase (ACP), ACP can hydrolyze AAP to produce antioxidative ascorbic acid (AA), inhibiting TMB chromogenic reaction. After that, introduction of As(V) can inhibit ACP, recovering TMB chromogenic reaction. Therefore, sensitive and selective As(V) detection is achieved. Moreover, Ce(IV)-ATP-Tris CPNs were transformed into cellulose nanofiber (CNF) to form test strip (Ce(IV)-ATP-Tris CPNs/CNF). Inorganic arsenic in rice can be detected by test strip, color of that can be measured by smartphone-integrated colorimetric quantitative analysis platform. Given this, rapid and convenient strip test of inorganic arsenic in rice by using this platform was achieved. Hence, this platform possesses great application potential in the field of inorganic arsenic in rice, even food safety.

Development of Diagnosis Model for Early Lung Nodules Based on a Seven Autoantibodies Panel and Imaging Features.

Background: There is increasing incidence of pulmonary nodules due to the promotion and popularization of low-dose computed tomography (LDCT) screening for potential populations with suspected lung cancer. However, a high rate of false-positive and concern of radiation-related cancer risk of repeated CT scanning remains a major obstacle to its wide application. Here, we aimed to investigate the clinical value of a non-invasive and simple test, named the seven autoantibodies (7-AABs) assay (P53, PGP9.5, SOX2, GAGE7, GUB4-5, MAGEA1, and CAGE), in distinguishing malignant pulmonary diseases from benign ones in routine clinical practice, and construct a neural network diagnostic model with the development of machine learning methods. Method: A total of 933 patients with lung diseases and 744 with lung nodules were identified. The serum levels of the 7-AABs were tested by an enzyme-linked Immunosorbent assay (ELISA). The primary goal was to assess the sensitivity and specificity of the 7-AABs panel in the detection of lung cancer. ROC curves were used to estimate the diagnosis potential of the 7-AABs in different groups. Next, we constructed a machine learning model based on the 7-AABs and imaging features to evaluate the diagnostic efficacy in lung nodules. Results: The serum levels of all 7-AABs in the malignant lung diseases group were significantly higher than that in the benign group. The sensitivity and specificity of the 7-AABs panel test were 60.7% and 81.5% in the whole group, and 59.7% and 81.1% in cases with early lung nodules. Comparing to the 7-AABs panel test alone, the neural network model improved the AUC from 0.748 to 0.96 in patients with pulmonary nodules. Conclusion: The 7-AABs panel may be a promising method for early detection of lung cancer, and we constructed a new diagnostic model with better efficiency to distinguish malignant lung nodules from benign nodules which could be used in clinical practice.

Combined analysis of C-reactive protein in pleural fluid and serum is effective in the differential diagnosis of exudative pleural effusions.

BACKGROUND: Exudative pleural effusion (EPE) is one of the common pleural manifestations of various diseases. Differential diagnosis of EPE is imperative clinically as it identifies different causes of EPE, thereby, enabling effective treatments. Thoracoscopy is a useful tool for differential diagnosis of EPE; however, some patients refuse thoracoscopic examination due to its invasive nature. In addition, the specificity and sensitivity of existing routine tests of EPE are unsatisfactory. Therefore, there is a great need to establish an effective method for the differential diagnosis of EPE. METHODS: This study was a single-institution retrospective analysis of diagnostic efficiency of C-reactive protein (CRP) and procalcitonin (PCT) between March 2018 and September 2018. A total of 87 patients diagnosed with EPE were enrolled. All participants underwent diagnostic thoracentesis. The EPE was examined using biochemical, routine, microbiological, and cytological methods. Pathological cytology detection was necessary for those suspected of malignant PE. Benign PE originates in patients with pneumonia, empyema, and tuberculosis. The levels of CRP and PCT in EPE and serum were measured before treatment. Correlation analysis and receiver-operating characteristic (ROC) curve analysis were conducted to determine the underlying relationship between levels of CRP and PCT, and for differential diagnosis. RESULTS: The ROC analysis showed that the sensitivity and specificity for the analysis of pleural fluid CRP (p-CRP) were higher (cut-off: 17.55 pg/mL; sensitivity: 75.00%, specificity: 83.90%) than that of serum CRP (s-CRP, cut-off: 23.90 pg/mL; sensitivity: 71.00%, specificity: 80.4%) in the differential diagnosis for EPE. However, the analysis of pleural fluid PCT (p-PCT) and serum PCT (s-PCT) did not demonstrate correlations with EPE. Combined analysis of p-CRP (cut-off: 17.55 mg/dL) with s-CRP (cut-off: 23.9 pg/mL) showed the highest diagnostic accuracy (88.4%) in diagnosing infectious EPE. CONCLUSIONS: The data support the close relationship between combined analysis of p-CRP with s-CRP and effective and accurate differential diagnosis of EPE, due to its higher sensitivity and specificity. However, as a highly sensitive marker for diagnosing bacterial infections, neither s-PCT nor p-PCT, showed correlations with the differential diagnosis of EPE.

Long non-coding RNA small nucleolar RNA host gene 6 aggravates pancreatic cancer through upregulation of far upstream element binding protein 1 by sponging microRNA-26a-5p.

BACKGROUND: Pancreatic cancer (PC) is a highly deadly malignancy with few effective therapies. We aimed to unmask the role that long non-coding RNA small nucleolar RNA host gene 6 (SNHG6) plays in PC cells by targeting far upstream element binding protein 1 (FUBP1) via microRNA-26a-5p (miR-26a-5p). METHODS: SNHG6 expression was predicted by bioinformatics, followed by verification via reverse transcription quantitative polymerase chain reaction. Then, the interactions among SNHG6, miR-26a-5p, and FUBP1 were detected through online software analysis, dual luciferase reporter assay and RNA pull-down. After that, cells were treated with different small interfering RNAs and/or mimic to determine the interactions among SNHG6, miR-26a-5p, and FUBP1 and their roles in PC cells. Finally, the role of SNHG6 in tumor growth in vivo was evaluated by measuring the growth and weight of transplanted tumors in nude mice. A t-test, one-way and two-way analysis of variance were used for data analysis. RESULTS: Compared with that in normal tissues, SNHG6 was highly expressed in PC tissues (1.00 ±â€Š0.05 vs. 1.56 ±â€Š0.06, t = 16.03, P < 0.001). Compared with that in human pancreatic duct epithelial cells (HPDE6-C7), SNHG6 showed the highest expression in PANC-1 cells (1.00 ±â€Š0.06 vs. 3.87 ±â€Š0.13, t = 34.72, P < 0.001) and the lowest expression in human pancreatic cancer cells (MIAPaCa-2) (1.00 ±â€Š0.06 vs. 1.41 ±â€Š0.07, t = 7.70, P = 0.0015). Compared with the levels in the si-negative control group, SNHG6 (0.97 ±â€Š0.05 vs. 0.21 ±â€Š0.06, t = 16.85, P < 0.001), N-cadherin (0.74 ±â€Š0.05 vs. 0.41 ±â€Š0.04, t = 8.93, P < 0.001), Vimentin (0.55 ±â€Š0.04 vs. 0.25 ±â€Š0.03, t = 10.39, P < 0.001), and ß-catenin (0.62 ±â€Š0.05 vs. 0.32 ±â€Š0.03, t = 8.91, P < 0.001) were decreased, while E-cadherin (0.65 ±â€Š0.06 vs. 1.36 ±â€Š0.07, t = 13.34, P < 0.001) was increased after SNHG6 knockdown or miR-26a-5p overexpression, accompanied by inhibited cell proliferation, migration, and invasion. SNHG6 overexpression exerted the opposite effects. SNHG6 upregulated FUBP1 expression by sponging miR-26a-5p. Silencing SNHG6 blocked the growth of PC in vivo. CONCLUSION: Silencing SNHG6 might ameliorate PC through inhibition of FUBP1 by sponging miR-26a-5p, thus providing further supporting evidence for its use in PC treatment.

Caveolin-1 Inhibits Proliferation, Migration, and Invasion of Human Colorectal Cancer Cells by Suppressing Phosphorylation of Epidermal Growth Factor Receptor.

BACKGROUND Although downregulation of caveolin-1 (Cav-1), which is a key constituent of membrane caveolae and a regulator of cellular processes, is associated with colorectal cancer (CRC), its involvement in the disease progression is largely unknown. This study aimed to explore the role of Cav-1 in CRC and the associated mechanisms. MATERIAL AND METHODS Fresh tissues from patients with CRC and human CRC SW480 cells were used to evaluate Cav-1 and Ki-67 expression using immunohistochemistry and Western blotting. The MTS and Transwell assays were performed to determine the effects of Cav-1 overexpression via pcDNA3.1/Cav-1 plasmid on cell proliferation and metastasis. The effect of Cav-1 on the epidermal growth factor receptor (EGFR) activation was evaluated by Western blotting. The correlation of Cav-1 expression with clinicopathological factors was statistically analyzed. RESULTS Overexpression of Cav-1 significantly reduced proliferation, migration, and invasion of SW480 cancer cells in vitro. The EGF-induced phosphorylation of EGFR and activations of the RAF-MEK-ERK and PI3K-AKT pathways were adversely regulated by Cav-1 overexpression in vitro. In 76 cases of CRC patients with EGFR expression, a negative correlation was observed between the level of Cav-1 and tumor-node-metastasis stage, lymph node metastasis, and distant metastasis (All p<0.05). Finally, the expression level of Cav-1 was negatively correlated with that of Ki-67. CONCLUSIONS This report is the first to show that overexpression of Cav-1significantly inhibits the proliferation, migration, and invasion potential of SW480 cells, possibly through reducing EGF-induced EGFR activation. High Cav-1 expression level may be a predictor of positive outcomes in patients with colorectal cancer.

Preliminary comparison of diffusion-weighted MRI and PET/CT in predicting histological type and malignancy of lung cancer.

INTRODUCTION: Emerging evidence shows that diffusion-weighted magnetic resonance imaging (DW MRI) and fluorine 18 fluorodeoxyglucose positron emission tomography/computed tomography (18 F-FDG PET/CT) might be useful in predicting histological type and malignancy of lung cancer, and even in specifically detecting the types of gene mutation. OBJECTIVE: We assessed whether DW MRI is equivalent to PET/CT in lung cancer diagnosis and evaluation. METHODS: The institutional review board approved this study, and written informed consent was obtained from all patients. DW MRI and FDG PET/CT were performed before therapy in 15 lung cancer patients diagnosed by pathological examination. Apparent diffusion coefficient (ADC), ratio of ADC (rADC = ADC in tumor/ADC in spinal cord) and maximal standardized uptake value (SUVmax ) were assessed. RESULTS: ADC, rADC and SUVmax did not reveal significant differences among different types of lung cancer. Sensitivity, specificity and accuracy of ADC, rADC and SUVmax proved to be not significantly different in the detection of adenocarcinoma and squamous cell carcinoma. Difference in the abilities of the sensitivity, specificity and accuracy of ADC, rADC and SUVmax to detect adenocarcinoma and squamous cell carcinoma proved to be insignificant. Although Ki-67 score did not show correlation with ADC, rADC and SUVmax , significant positive correlation was found between ADC and rADC, and ADC and SUVmax . CONCLUSIONS: Both DW MRI and FDG PET/CT had similar limited diagnostic capability of predicting different histological types and malignancy of lung cancer. This study may help provide a novel insight into diagnostic and therapeutic strategies of lung cancer based on DW MRI.

Effect of overexpression of hypoxia-inducible factor-1α induced by hyperoxia in vivo in LNCaP tumors on tumor growth rate.

OBJECTIVE: To study effect of overexpression of hypoxia-inducible factor-1α induced by hyperoxia in vivo in LNCaP tumors on tumor growth rate. METHODS: The prostate cancer LNCaP cells were inoculated in the abdomen of mice. All the mice were randomly placed in the gas chamber with different oxygen content. The groups were divided as follows: twelve mice in hypoxia group, sixteen mice in normoxia group, ten mice in hyperoxia group. After 28 d of treatment, the mice were weighed, the blood samples were taken from the left ventricle, and the tumor was isolated and weighed. Tumor growth, angiogenesis and vascularization, HIF-1α expression and intracellular signal transduction molecules expression in each group of xenografts were detected and analyzed by using Western blotting and immunofluorescence and determination of hemoglobin. RESULTS: Comparison of the growth of xenografts in each group showed that, the xenografts growth of hypoxia group was more quickly than that of normoxia group. The difference was statistically significant (P = 0.004). The difference in xenografts growth between hyperoxia group compared and normoxia group was not statistically significant (P > 0.05). The expressions of HIF-1α, VEGF and VEGF-R of xenografts in hyperoxia group were significantly higher than those of normoxia group (P < 0.05). The expression of HIF-1α of xenografts in hypoxia group and normoxia group were similar. The blood growth rate of xenografts in hypoxia group (170%) was significantly higher than that of normoxia group (40%) (P < 0.05). The expression of Nrf2 of xenografts in hyperoxia group was significantly higher than that of normoxia group (P < 0.05). CONCLUSIONS: When hyperoxia induces the overexpression of HIF-1α in LNCaP tumor, it will not affect tumor growth. It provides a new ideas and theoretical basis for the clinical treatment of prostate cancer.