Antiparasitic efficacy of flavonoids identified from Psoralea corylifolia against Tetrahymena piriformis in guppy (Poecilia reticulate).

Tetrahymena piriformis belongs to the ciliated protists (ciliates), causing severe economic losses in aquaculture. Chemical drugs currently used usually have toxic side effects, and there is no specific drug against Tetrahymena. Therefore, it is an urgent need to identify new antiparasitic lead compounds. In the present study, the in vitro parasiticidal activity of ethyl acetate (EtOAc) extracts and water extracts from 22 selected traditional Chinese medicines (TCMs) were evaluated against T. piriformis. The EtOAc extract of P. corylifolia turned out to be the most active with the minimum parasiticidal concentration of 100 mg/L within 3 h. Thus, it was separated into 12 fractions by the first-dimensional (D1) normal phase liquid chromatography (NPLC), meanwhile combining with in vitro antiparasitic tests for activity tracking. Subsequently, 8 flavonoids were identified in the active fractions by the second-dimensional (D2) reverse phase liquid chromatography (RPLC) tandem high-resolution mass spectrometry. According to the results, 5 flavonoids were selected for in vitro antiparasitic test, of which isobavachalcone showed the minimum parasiticidal concentration of 3.125 mg/L in 2 h. Bathing treatment of infected guppies with isobavachalcone could significantly reduce the burden of T. piriformis, obtaining a 24-h median effective concentration (24-h EC50) value of 1.916 mg/L. And the concentration of isobavachalcone causing guppies to die within 24 h is 39 times than that of 24-h EC50. The results demonstrated that isobavachalcone has the potential to be developed into a novel commercial fish drug against T. piriformis.

Circ_0004104 participates in the regulation of ox-LDL-induced endothelial cells injury via miR-942-5p/ROCK2 axis.

BACKGROUND: Cardiovascular disease was the most common disease among the elderly with high morbidity and mortality. Circ_0004104 was demonstrated to be involved in the regulation of atherosclerosis. METHODS: Quantitative real-time polymerase chain reaction was employed to measure the expression of circ_0004104, miR-942-5p and Rho associated coiled-coil containing protein kinase 2 (ROCK2). Cell proliferation was tested by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Cell apoptosis was measured by flow cytometry, and tube formation assay was used to detect the angiogenesis ability of cells. Western blot assay was performed to assess protein levels. Enzyme­linked immunosorbent assay was used to detect the release of IL-1ß and TNF-α. The relationship between miR-942-5p and circ_0004104 or ROCK2 was identified by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay, and RNA pull-down assay. RESULTS: Oxidized low-density lipoprotein (ox-LDL) inhibited the proliferation of human umbilical vein endothelial cells (HUVECs) and promoted apoptosis in a dose-dependent manner. Circ_0004104 was increased in serum of atherosclerosis patients and ox-LDL-treated HUVECs, and silence of circ_0004104 promoted the proliferation of ox-LDL-exposed HUVECs and inhibited cell apoptosis. MiR-942-5p downregulation reversed si-circ_0004104-mediated influences in HUVECs upon ox-LDL exposure. ROCK2 was the target of miR-942-5p and circ_0004104 regulated the expression of ROCK2 through sponging miR-942-5p. ROCK2 abated the influences of miR-942-5p in ox-LDL-stimulated HUVECs. Circ_0004104 was increased in the exosomes derived from ox-LDL-exposed HUVECs, and the expression of circ_0004104 was promoted in HUVECs after stimulation with ox-LDL-treated HUVECs cells-derived exosomes. CONCLUSION: Circ_0004104 downregulation receded ox-LDL-induced injury in HUVECs through miR-942-5p and ROCK2.

First case of Coxiella burnetii infection in Palaemonetes sinensis cultured in Liaoning, China.

The Palaemonetes sinensis aquaculture industry in Panjin City, Liaoning Province, China, experienced heavy losses in October 2018. Morbidity of cultured shrimp reached 50% and was characterized by cloudiness of muscle and the gradual spread of disease within the population. When the infection was mild, histopathological examinations revealed that the muscle cells contained a considerable number of microorganisms. In extreme cases, the structure of the hepatopancreatic glandular and muscle fiber was obscured or even vanished. Electron microscope observations revealed the presence of granular cytoplasmic inclusions in cells from hepatopancreas and muscle tissues. The 16S rDNA sequence of the intracytoplasmic organism was 94.7% identity to that of Coxiella burnetii. This is the first report of infection by C. burnetii in P. sinensis.

Metschnikowia bicuspidata isolated from milky diseased Eriocheir sinensis: Phenotypic and genetic characterization, antifungal susceptibility and challenge models.

A severe disease occurred in farmed Eriocheir sinensis characterized by milky liquid accumulation in the pectoral cavity, in the province of Liaoning, China, during October 2018-April 2019. Diseased crabs moved sluggishly, exhibited appetite loss and readily lost legs. Under the microscopic analysis, it was observed that the milky liquid contained a large number of yeastlike microorganisms (0.8-1.2 µm × 1.5-1.9 µm), which were also present in the muscle, hepatopancreas and gills. A dominant strain was isolated from the milky liquid and other tissues of diseased crabs. It grew on nutrient agar and formed 1- to 3-mm white opaque colonies, each with a protuberance in the centre. Besides, the results of TEM and SEM also demonstrate a typical multilateral budding model of the yeast clearly. We identified the strain, which we named 2EJM001, as Metschnikowia bicuspidata based on 18S rDNA, ITS and 26S rDNA sequence analyses and on its morphological, physiological and biochemical characteristics. Phylogenetic analysis revealed that 26S rDNA of 2EJM001 was clustered with M. bicuspidata (LNES0119) as reported by Bao et al. In addition, unlike Bao et al., two challenge experiments (injection and immersion) were used in this study. The results of challenge experiments show that 2EJM001 was pathogenic to E. sinensis and caused signs similar to those found in the naturally infected crabs. At the same time, the minimum inhibitory concentrations (MIC80 and MIC90 ) were determined. This study further confirms that M. bicuspidata 2EJM001 was the pathogen responsible for 'milky disease' in E. sinensis from Liaoning Province.

First case of non-O1/O139 Vibrio cholerae infection in the pink-tailed chalceus Chalceus macrolepidotus in China.

A pathogen was isolated from diseased pink-tailed chalceus Chalceus macrolepidotus during a high-mortality outbreak in a freshwater culture farm in Liaoyang, China. The diseased fish were characterized by disoriented behaviors, exophthalmos, and redness and swelling of the top of the head. A Gram-negative, pure strain of bacteria (CM0428) was isolated from the brain, kidney, and liver. The isolate was identified as Vibrio cholerae based on ompW gene amplification and 16S rRNA and gyrB gene sequence analysis. Serogroup testing indicated that CM0428 was a non-O1/O139 strain of V. cholerae. The challenge test showed that CM0428 exhibited strong virulence to pink-tailed chalceus, and hlyA and toxR virulence-related genes were detected. The isolate was sensitive to multi-class antibiotics, but resistant to tetracycline. Histological examination revealed that V. cholerae CM0428 infection caused multiple organ and tissue lesions, and typical pathological features were cell degeneration and necrosis.

A Low-Cost, Ear-Contactless Electronic Stethoscope Powered by Raspberry Pi for Auscultation of Patients With COVID-19: Prototype Development and Feasibility Study.

BACKGROUND: Chest examination by auscultation is essential in patients with COVID-19, especially those with poor respiratory conditions, such as severe pneumonia and respiratory dysfunction, and intensive cases who are intubated and whose breathing is assisted with a ventilator. However, proper auscultation of these patients is difficult when medical workers wear personal protective equipment and when it is necessary to minimize contact with patients. OBJECTIVE: The objective of our study was to design and develop a low-cost electronic stethoscope enabling ear-contactless auscultation and digital storage of data for further analysis. The clinical feasibility of our device was assessed in comparison to a standard electronic stethoscope. METHODS: We developed a prototype of the ear-contactless electronic stethoscope, called Auscul Pi, powered by Raspberry Pi and Python. Our device enables real-time capture of auscultation sounds with a microspeaker instead of an earpiece, and it can store data files for later analysis. We assessed the feasibility of using this stethoscope by detecting abnormal heart and respiratory sounds from 8 patients with heart failure or structural heart diseases and from 2 healthy volunteers and by comparing the results with those from a 3M Littmann electronic stethoscope. RESULTS: We were able to conveniently operate Auscul Pi and precisely record the patients' auscultation sounds. Auscul Pi showed similar real-time recording and playback performance to the Littmann stethoscope. The phonocardiograms of data obtained with the two stethoscopes were consistent and could be aligned with the cardiac cycles of the corresponding electrocardiograms. Pearson correlation analysis of amplitude data from the two types of phonocardiograms showed that Auscul Pi was correlated with the Littmann stethoscope with coefficients of 0.3245-0.5570 for healthy participants (P<.001) and of 0.3449-0.5138 among 4 patients (P<.001). CONCLUSIONS: Auscul Pi can be used for auscultation in clinical practice by applying real-time ear-contactless playback followed by quantitative analysis. Auscul Pi may allow accurate auscultation when medical workers are wearing protective suits and have difficulties in examining patients with COVID-19. TRIAL REGISTRATION: ChiCTR.org.cn ChiCTR2000033830; http://www.chictr.org.cn/showproj.aspx?proj=54971.

Toward a national emission inventory for the catering industry in China.

Catering oil fumes are a major hazard to human health. In particular, the typical Chinese cooking style is characterised by a high temperature frying process that produces high levels of cooking oil fumes. However, limited data relating to this sector mean that national emission inventory data specific to the catering service industry do not exist. To address above deficiency and thus to establish the inventory of a city, or a province, or even a country, a door-to-door survey campaign was launched in the Chinese cities of Heze and Linfen to determine the structure of local catering industries. Data revealed that the number of catering businesses per 104 people was 17 ± 4. Of these, 3.0 ± 1.4, 15.0 ± 1.4, and 82.0 ± 0.0% were classified as large, medium, and small enterprises, respectively. Furthermore, the installation rates of fume purifiers were 74 ± 13, 66 ± 9, and 51 ± 14% for large, medium, and small enterprises, respectively, with net removal efficiencies of 63 ± 11, 50 ± 7, and 31 ± 8%, respectively. This information was extrapolated across all provincial regions of China to construct a provincial and national emission inventory. In 2017, China's national catering industry released approximately 34 kt of volatile organic compounds (VOCs), 38 kt of particulate matter with a diameter less than 2.5 µm (PM2.5), 48 kt of particulate matter with a diameter less than 10 µm (PM10), 1 kt of black carbon (BC), and 27 kt of organic carbon (OC). A significant correlation was observed between vegetable oil consumption and emissions (e.g., for VOCs, y = 14.94 x + 76.50, R2 = 0.87, where y is VOCs emissions and x is vegetable oil consumption), indirectly corroborating the rationality of the inventory. Moreover, this correlation provides the potential for a dynamic inventory based on vegetable oil consumption. Future studies are proposed to address more influential factors to improve the reliability of the national inventory and refer to big data, rather than door-to-door investigation, to identify the amount of catering service businesses in a region.

Algorithm developed for dynamic quantification of coal consumption for and emission from rural winter heating.

Coal-dominated winter heating practices in China are largely accepted to be a leading cause of winter haze in the region though the amount of coal for heating is actually much lower than for power generation or industrial process. However, little is known about how the total rural coal weight in a region could be attributed to real time (e.g., daily) patterns, limiting the understanding of dynamic impacts of coal emissions and the adoption of timely measures against predicted haze. Considering that winter heating essentially protects against cold temperatures, coal burning strength may be related to the temperatures that people experience. A field study was organized to test the validity of this hypothesis. A system was designed to continuously monitor every instance of coal addition, and coal consumption on any given day for a whole village (WDAY) was calculated by summating all the additions. Meanwhile, a new term, composite temperature (TCOM), which incorporates a few weather-related elements, was introduced to represent cold temperatures that individuals experience. It was found that WDAY and TCOM presented opposite variations, and a negative linear correlation was observed (WDAY = -0.75TCOM + 11.86, R2 = 0.75), revealing the feasibility of estimating coal consumption on a certain day (WDAY) based on weather data (TCOM) for a given village. An extensive form of the algorithm for any area of interest (e.g., a district, city, or province) can be expressed as WDAY = (-0.75TCOM + 11.86)‧NH/834, where NH denotes the number of households in a region. This algorithm reflects the essence of winter heating (to resist cold temperatures), and therefore its logic is highly likely to be useful for any countries of the world regardless of what forms of energy used (coal or other energy forms) provided the energy involved is unexceptionally used for winter heating, though there may be some uncertainties in estimated coal consumption due to multiple factors.

Study on Hesitant Fuzzy Information Measures and Their Clustering Application.

At present, research on hesitant fuzzy operations and measures is based on equal length processing, and an equal length processing method will inevitably destroy the original data structure and change the data information. This is an urgent problem to be solved in the development of hesitant fuzzy sets. Aiming at solving this problem, this paper firstly defines a hesitant fuzzy entropy function as the measure of the degree of uncertainty of hesitant fuzzy information and then proposes the concept of hesitant fuzzy information feature vector. The hesitant fuzzy distance measure and similarity measure are studied based on the information feature vector. Finally, the hesitant fuzzy network clustering method based on similarity measure is given, and the effectiveness of our algorithm through a numerical example is illustrated.

Ranolazine improves oxidative stress and mitochondrial function in the atrium of acetylcholine-CaCl2 induced atrial fibrillation rats.

AIMS: Ranolazine, an antianginal agent used for chronic stable angina treatment, was demonstrated to be effective in atrial fibrillation (AF) treatment. The aim of this study was to explore the molecular mechanisms of its anti-AF effects. MAIN METHODS: AF rat model was established using acetylcholine (ACh)-CaCl2 injection for 7days followed by ACh infusion into the heart. Prior to ACh infusion, ranolazine at 10.7mg/kg/0.5ml was injected into vein and followed by 0.56mg/kg/min infusion. Blood pressure and electrocardiogram were monitored during the infusion. Histological changes of atrial tissue were observed after H&E staining. Activities and protein expression of NADPH oxidase-4, xanthine oxidase, glutathione peroxidase and superoxide dismutase were examined using commercial assay kits and Western botting, respectively. Mitochondrial functions were evaluated through membrane potential, ATP production, activities of complex I and III and reactive oxygen species production. Apoptosis was measured using TUNEL staining. Protein expression of apoptotic proteins Bcl-2, Bax and cleaved-caspase 3 and Akt/mTOR signaling proteins were detected using Western blotting. KEY FINDINGS: Results demonstrated that ranolazine attenuated AF in ACh-CaCl2-exposed rats. In addition, ranolazine restored mitochondrial function, suppressed oxidative stress, and inhibited atrial cells apoptosis. Furthermore, the activated Akt/mTOR signaling pathway induced by AF was further activated by ranolazine. SIGNIFICANCE: The present study confirms the effects of ranolazine on AF rats induced by ACh-CaCl2, and provides evidence that the anti-AF effects are associated with the restoration of mitochondrial function and activation of the Akt/mTOR signaling pathway in atrial tissue.