NPM promotes hepatotoxin-induced fibrosis by inhibiting ROS-induced apoptosis of hepatic stellate cells and upregulating lncMIAT-induced TGF-ß2.

Liver fibrosis is caused by a variety of chronic liver injuries and has caused significant morbidity and mortality in the world with increasing tendency. Elucidation of the molecular mechanism of liver fibrosis is the basis for intervention of this pathological process and drug development. Nucleophosmin (NPM) is a widely expressed nucleolar phosphorylated protein, which is particularly important for cell proliferation, differentiation and survival. The biological role of NPM in liver fibrosis remains unknown. Here we show that NPM promotes liver fibrosis through multiple pathways. Our study found that NPM was up-regulated in cirrhosis tissues and activated in hepatic stellate cells (HSCs). NPM inhibition reduced liver fibrosis markers expression in HSCs and inhibited the HSCs proliferation and migration. In mice model, NPM knockdown in HSCs or application of specific NPM inhibitor can remarkably attenuate hepatic fibrosis. Mechanistic analysis showed that NPM promotes hepatic fibrosis by inhibiting HSCs apoptosis through Akt/ROS pathway and by upregulating TGF-ß2 through Akt-induced lncMIAT. LncMIAT up-regulated TGF-ß2 mRNA by competitively sponging miR-16-5p. In response to liver injury, hepatocytes, Kupffer cells and HSCs up-regulated NPM to increase TGF-ß2 secretion to activate HSCs in a paracrine or autocrine manner, leading to increased liver fibrosis. Our study demonstrated that NPM regulated hepatotoxin-induced fibrosis through Akt/ROS-induced apoptosis of HSCs and via the Akt/lncMIAT-up-regulated TGF-ß2. Inhibition of NPM or application of NPM inhibitor CIGB300 remarkably attenuated liver fibrosis. NPM serves a potential new drug target for liver fibrosis.

Erratum: Jiang et al. Localization Accuracy of Ultrasound-Actuated Needle with Color Doppler Imaging. Diagnostics 2020, 10, 1020.

The authors wish to correct the following erratum in this paper [...].

Development of high frequency piezocomposite with hexagonal pillars via cold ablation process.

This paper reports on the fabrication of 1-3 piezocomposite with hexagonal pillars for high frequency ultrasonic transducer based on the cold ablation technique. The piezocomposite with hexagonal pillars was designed, simulated, and fabricated using an ultraviolet picosecond laser. It performs better than the piezocomposite with other pillar shapes like square. The edge length and height of the hexagonal PZT pillar were 10 µm and 36 µm, the width of the kerf was about 5 µm. The 1-3 piezocomposite with a resonance frequency of 51.2 MHz and a coupling coefficient of 0.69 was fabricated. The transducer with fabricated 1-3 piezocomposite was prototyped and characterized. Compared to the conventional dice-and-fill technique, the cola ablation process allows for the manufacturing of 1-3 piezocomposites with higher variability of pillar design and distribution as well as smaller structural size. It suggests that the cold ablation process proves to be suitable for the fabrication of high frequency composite and transducers.

Micromachined High Frequency 1-3 Piezocomposite Transducer Using Picosecond Laser.

In this article, a PZT/Epoxy 1-3 piezoelectric composite based on picosecond laser etching technology is developed for the fabrication of high-frequency ultrasonic transducer. The design, fabrication, theoretical analysis, and performance of the piezocomposite and transducer are presented and discussed. According to the test results, the area of the PZT pillar is [Formula: see text], the average width of the kerf is [Formula: see text], and the thickness of the piezocomposite is [Formula: see text]. The fabricated 1-3 piezocomposite has a resonant frequency of 46.5 MHz, a parallel resonant frequency of 65 MHz, and an electromechanical coupling coefficient of 0.73. According to the wires phantom imaging, its imaging resolution can reach [Formula: see text]. This study shows that the proposed picosecond laser micromachining technique can be applied in the fabrication of high frequency 1-3 piezocomposite and transducer.

HnRNPA2B1 promotes the proliferation of breast cancer MCF-7 cells via the STAT3 pathway.

HnRNPA2/B1 is highly expressed in many tumors. However, the role of hnRNPA2/B1 in breast cancer is not clear. In this study, we found the proliferation rate was decreased after knockout of hnRNPA2/B1 by CRISPR-CAS9 in MCF-7 cells, as demonstrated by the reduced expression of CDK4 and p-AKT, and the increased expression of P27. Besides this, the western blot results showed that knockout of hnRNPA2/B1 increased the rate of apoptosis and declined autophagy. By in vivo assay, we found that knockout of hnRNPA2/B1 suppressed tumor growth in a xenograft mouse model. Immunohistochemical staining results confirmed knockout of hnRNPA2/B1 impaired tumor angiogenesis, as illustrated by downregulated expression of VEGF-A. Besides this, interacting proteins with hnRNPA2/B1 were identified by mass spectrometry and the PPI network was constructed. GO analysis suggests that the Interacting proteins are mainly enriched in the Wnt signaling pathway, tumor necrosis factor-mediated signaling pathway, translation, and so on. We then identified hnRNPA2/B1 interacted with signal transducer and activator of transcription 3 (STAT3), as supported by the colocalization of hnRNPA2/B1 and STAT3. Meanwhile, knockout of hnRNPA2/B1 inhibited the phosphorylation of STAT3. Collectively, our results demonstrate that hnRNPA2/B1 promotes tumor cell growth in vitro and in vivo by activating the STAT3 pathway, regulating apoptosis and autophagy.

Localization Accuracy of Ultrasound-Actuated Needle with Color Doppler Imaging.

An ultrasonic needle-actuating device for tissue biopsy and regional anaesthesia offers enhanced needle visibility with color Doppler imaging. However, its specific performance is not yet fully determined. This work investigated the influence on needle visibility of the insertion angle and drive voltage, as well as determined the accuracy and agreement of needle tip localization by comparing color Doppler measurements with paired photographic and B-mode ultrasound measurements. Needle tip accuracy measurements in a gelatin phantom gave a regression trend, where the slope of trend is 0.8808; coefficient of determination (R2) is 0.8877; bias is -0.50 mm; and the 95% limits of agreement are from -1.31 to 0.31 mm when comparing color Doppler with photographic measurements. When comparing the color Doppler with B-mode ultrasound measurements, the slope of the regression trend is 1.0179; R2 is 0.9651; bias is -0.16 mm; and the 95% limits of agreement are from -1.935 to 1.605 mm. The results demonstrate the accuracy of this technique and its potential for application to biopsy and ultrasound guided regional anaesthesia.

Sensitivity Enhanced Photoacoustic Imaging Using a High-Frequency PZT Transducer with an Integrated Front-End Amplifier.

Photoacoustic (PA) imaging is a hybrid imaging technique that can provide both structural and functional information of biological tissues. Due to limited permissible laser energy deposited on tissues, highly sensitive PA imaging is required. Here, we developed a 20 MHz lead zirconium titanate (PZT) transducer (1.5 mm × 3 mm) with front-end amplifier circuits for local signal processing to achieve sensitivity enhanced PA imaging. The electrical and acoustic performance was characterized. Experiments on phantoms and chicken breast tissue were conducted to validate the imaging performance. The fabricated prototype shows a bandwidth of 63% and achieves a noise equivalent pressure (NEP) of 0.24 mPa/√Hz and a receiving sensitivity of 62.1 µV/Pa at 20 MHz without degradation of the bandwidth. PA imaging of wire phantoms demonstrates that the prototype is capable of improving the detection sensitivity by 10 dB compared with the traditional transducer without integrated amplifier. In addition, in vitro experiments on chicken breast tissue show that structures could be imaged with enhanced contrast using the prototype and the imaging depth range was improved by 1 mm. These results demonstrate that the transducer with an integrated front-end amplifier enables highly sensitive PA imaging with improved penetration depth. The proposed method holds the potential for visualization of deep tissue structures and enhanced detection of weak physiological changes.

Preparation, characterization, and in vivo evaluation of a polymorphic form of valnemulin hydrogen tartrate.

We prepared a polymorphic form of valnemulin hydrogen tartrate (Form I) to overcome the instability and irritating odor of valnemulin hydrochloride that affect its use in the production and application of veterinary drugs. The physicochemical properties of Form I were characterized by scanning electron microscopy, X-ray powder diffraction, infrared spectroscopy, differential scanning calorimetry, and thermogravimetric analysis. The results showed the crystal structure and thermal properties of Form I were very different from those of a commercially available form of valnemulin hydrogen tartrate (Form II). Form I and Form II were more stable than valnemulin hydrochloride after storage under irradiation and high humidity conditions, respectively. The solubility of Form I was 2.6 times that of Form II, and Form I was selected for use in pharmaceutical kinetics experiments in vivo. Compared to valnemulin hydrochloride, after oral administration at a dose of 10 mg/kg in pigs, Form I had similar pharmaceutical kinetic behavior but a slightly higher area under the concentration-time curve from time zero to the last measurable concentration. Consequently, Form I should be suitable for the development of simple formulations and be effective in the clinical application of veterinary drugs.

Preparation of valnemulin hydrogen fumarate and its enhanced stability compared with valnemulin hydrochloride.

CONTEXT: It is necessary to develop a new salt of valnemulin to replace the veterinary antibiotic, e.g. valnemulin hydrochloride, in order to overcome its instability during storage and preparation. OBJECTIVE: The objective of this study was to prepare a novel organic acid salt, valnemulin hydrogen fumarate, and to investigate its stability compared with valnemulin hydrochloride. MATERIALS AND METHODS: The crystal of valnemulin hydrogen fumarate was prepared by modified crystallization method; the enhanced stabilities of valnemulin hydrogen fumarate were conducted under irradiation and humid conditions, and the experimental results were simulated at AM1 level of calculations. RESULTS: Valnemulin hydrogen fumarate was more stable than valnemulin hydrochloride. After irradiation for 180 days, the content of valnemulin hydrogen fumarate decreased slightly 2.7%, whereas the content of valnemulin hydrochloride had an obvious decrease of 32.8%. Meanwhile, valnemulin hydrogen fumarate showed better anti-RH (relative humidity) ability than valnemulin hydrochloride. Under conditions of 65% and 85% RH, the absorption values of valnemulin hydrogen fumarate towards water were 0.75% and 1.20% at 48 h, whereas those of valnemulin hydrochloride were 4.50% and 9.71%, respectively. CONCLUSION: The enhanced stability of valnemulin hydrogen fumarate could be attributed to its good crystallinity in comparison with the amorphous valnemulin hydrochloride.

Simultaneous determination of benzimidazoles and their metabolites in plasma using high-performance liquid chromatography/tandem mass spectrometry: application to pharmacokinetic studies in rabbits.

An HPLC/MS/MS method was developed for the simultaneous determination of the following benzimidazole anthelmintics and metabolites in plasma: flubendazole, albendazole, fenbendazole, mebendazole, thiabendazole, hydrolyzed flubendazole, albendazole sulfoxide, albendazole sulfone, albendazole aminosulfone, oxfendazole, fenbendazole sulfone, aminomebendazole, hydroxymebendazole, and 5-hydroxythiabendazole. The sample preparation process involved a pH-dependent extraction of the analytes. Chromatographic separation was performed on a C18 column with a mobile phase gradient starting with methanol-water (20 + 80, v/v) containing 0.1% formic acid. The overall average recoveries of the analytes based on a matrix-matched calibration ranged from 75.0 to 120.0%, with RSD values of <20.0%. The LODs ranged from 0.08 to 2.0 microg/kg and the LOQs from 0.3 to 5.0 microg/kg. The validated method was used in pharmacokinetic studies of benzimidazole compounds in rabbits, and the elimination of the metabolites was measured quantitatively.

[Determination of residues of cyromazine and its metabolite melamine in chickens by gas chromatography-mass spectrometry].

A gas chromatography-mass spectrometric (GC-MS) method has been established for the determination of cyromazine and its metabolite melamine in chickens. The homogenized tissue samples added with melamine-15N3 were extracted with acidic acetonitrile-water solution, and defatted with dichloromethane. The samples were derivatized with N,O-bis(trimethylsilyl) trifluoroacetamide (BSTFA), after solid-phase extraction (SPE) which was performed on Oasis MCX cartridges, and then detected by GC-MS. Cyromazine was quantified by an external standard method, and melamine by an internal standard method using melamine-15N3. The results indicated that the linearities of cyromazine and melamine were within the range of 100 - 1,000 microg/L, and their limits of quantification (LOQ) were 20 microg/kg. The mean recoveries of chicken samples fortified at three concentrations of 20, 40 and 80 microg/kg were within the range of 75% - 110%. The relative standard deviations (RSDs) of intra- and inter- batch were less than 10% and 15%, respectively. The application of this method was further demonstrated by analyzing 10 real samples which were brought from local markets. The results show that this method is simple, rapid, sensitive and specific. It is appropriate for the identification and quantification of cyromazine and its metabolite melamine in chickens.

Determination of residues of cyromazine and its metabolite, melamine, in animal-derived food by gas chromatography-mass spectrometry with derivatization.

A gas chromatography-mass spectrometric (GC-MS) method was established for the determination of cyromazine and its metabolite, melamine, in animal-derived food. Chicken and tilapia muscle samples were spiked with (15)N(3)-melamine, extracted with an acidic acetonitrile/water solution, and defatted with dichloromethane. Egg and milk samples were directly extracted with 3% trichloroacetic acid. The extracts were purified using mixed cation-exchange cartridges, derived with N,O-bis(trimethylsilyl)trifluoroacetamide, and detected by GC-MS. Cyromazine and melamine were quantified by external standard methods except for the determination of melamine in animal muscle, which used an internal standard method. Recoveries ranged from 75.0 to 110.0%, and relative standard deviations were <15.0%. In animal muscle the limits of quantification (LOQs) were 20 microg/kg and the limits of detection (LODs) were 10 microg/kg for cyromazine and melamine. In milk and eggs the LOQs were 10 microg/kg and the LODs were 5 microg/kg for both analytes.

Photocatalytic degradation of pesticide pyridaben. 3. In surfactant/TiO2 aqueous dispersions.

The effective TiO2 photocatalytic degradation of pyridaben in an acetronitrile/water dispersion has been investigated in previous work, but could not be achieved in the case of real waters. In this paper, photocatalytic degradation of pyridaben on TiO2 particles under UV light (lambda> 360 nm) illumination in surfactant CTAB (cetyltrimethyl ammonium bromide) aqueous dispersions was studied. 1H NMR was used to determine quantitative information about the adsorption mode of pyridaben in CTAB micelles. The results showed that the upfield 1H shifts were largest for long chain protons of CTAB, indicating that the hydrophobic aromatic rings were primarily located in this region. Adsorption models on TiO2 surface were thus proposed. The reaction rates decreased with the increase of pH value, which can be attributed to the surface charge variations of pyridaben adsorbed onto TiO2 particles. The adsorption isotherms at different pH values confirmed that preadsorption on the surface of TiO2 particles was prerequisite for efficient degradation. Furthermore, an oxidation reagent such as H2O2 was added to the photocatalytic system, which may act as an alternative electron acceptor and result in a notably enhanced rate of pollutant destruction. On the basis of intermediates identified by GC/MS, a degradation pathway was proposed.