Prevalence and molecular characterization of multi-resistant Escherichia coli isolates from clinical bovine mastitis in China.

Different antibiotics are used to treat mastitis in dairy cows that is caused by Escherichia coli (E. coli). Antimicrobial resistance in food-producing animals in China has been monitored since 2000. Surveillance data have shown that the prevalence of multiresistant E. coli in animals has increased significantly. This study aimed to investigate the occurrence and molecular characteristics of resistance determinants in E. coli strains (n = 105) obtained from lactating cows with clinical bovine mastitis (CBM) in China. A total of 220 cows with clinical mastitis, which has swollen mammary udder with reduced and red or gangrenous milk, were selected from 5000 cows. The results showed 94.3% of the isolates were recognized as multidrug resistant. The isolates (30.5%) were positive for the class I integrase gene along with seven gene cassettes that were accountable for resistance to trimethoprim resistance (dfrA17, dfr2d and dfrA1), aminoglycosides resistance (aadA1 and aadA5) and chloramphenicol resistance (catB3 and catB2), respectively. The blaTEM gene was present in all the isolates, and these carried the blaCTX gene. A double mutation in gyrA (i.e., Ser83Leu and Asp87Asn) was observed in all fluoroquinolone-resistant isolates. In total, nine fluoroquinolone-resistant E. coli isolates were identified with five different types of mutations in parC. In four (44.4%) isolates, Ser458Ala was present in parE, and in all nine (9/9) fluoroquinolone-resistant isolates, Pro385Ala was present in gyrB. Meanwhile, fluoroquinolone was observed as highly resistant, especially in isolates with gyrA and parC mutations. In summary, the findings of this research recognize the fluoroquinolone resistance mechanism and disclose integron prevalence and ESBLs in E. coli isolates from lactating cattle with CBM.

Identification, cleavage, and amplification (ICA): A versatile strategy for highly sensitive detection of miRNA.

MicroRNAs (miRNAs) are small RNA molecules that can play important roles as diagnostic/prognostic biomarkers and therapeutic targets for cancers and other diseases. Herein, an identification-cleavage-amplification (ICA) strategy for highly sensitive and versatile detection of miRNA has been proposed, and successfully applied to miR-155 and miR-21 assays. It combines an aligner-target mediated cleavage with strand displacement amplification (ATMC-SDA) to achieve the ICA process. During the identification process, a DNA-aligner (DA) and a DNA-amplicon (DM) can bind together with the help of target miRNA, forming a T-junction structure. Then, a nicking endonuclease (NEase), binding on the recognition sequence at the stem part of DA, can make a cleavage on DM, and the cleaved DM (CDM) can serve as an initiator to trigger the SDA reaction for signal amplification. Sharing the same set of enzymes and primers, the proposed ATMC-SDA can serve as a versatile ICA strategy for highly sensitive detection of various miRNAs, without the requirement of reverse transcription. Results show that the limits of detection (LOD) for miR-155 and miR-21 are 5.4 aM and 6.8 aM, respectively, with a dynamic range from 10.0 aM to 10.0 pM. The compatibility of ATMC-SDA with biological samples has also been tested by using human serum, indicating a promising potential for a wide variety of applications.

A Sign Language Recognition System Applied to Deaf-Mute Medical Consultation.

It is an objective reality that deaf-mute people have difficulty seeking medical treatment. Due to the lack of sign language interpreters, most hospitals in China currently do not have the ability to interpret sign language. Normal medical treatment is a luxury for deaf people. In this paper, we propose a sign language recognition system: Heart-Speaker. Heart-Speaker is applied to a deaf-mute consultation scenario. The system provides a low-cost solution for the difficult problem of treating deaf-mute patients. The doctor only needs to point the Heart-Speaker at the deaf patient and the system automatically captures the sign language movements and translates the sign language semantics. When a doctor issues a diagnosis or asks a patient a question, the system displays the corresponding sign language video and subtitles to meet the needs of two-way communication between doctors and patients. The system uses the MobileNet-YOLOv3 model to recognize sign language. It meets the needs of running on embedded terminals and provides favorable recognition accuracy. We performed experiments to verify the accuracy of the measurements. The experimental results show that the accuracy rate of Heart-Speaker in recognizing sign language can reach 90.77%.

Structural Characteristics of Insoluble Dietary Fiber from Okara with Different Particle Sizes and Their Prebiotic Effects in Rats Fed High-Fat Diet.

Dietary fiber, which is utilized to make functional meals, is an important component for promoting human health and managing calorie consumption. In this study, three different particle sizes of OIDF (Okara insoluble dietary fiber) were characterized. Their lipid-lowering effects and the impacts on gut microbiota were determined by OIDF intervention in high-fat diet rats. Scanning electron microscopy (SEM) results showed that the three particle sizes of OIDF have different morphologies. Fourier transform infrared spectroscopy (FT-IR) results showed that the three sources of IDF samples have similar active groups, but the thermogravimetric analysis/differential scanning calorimetry (TGA/DSC) and X-ray diffraction (XRD) showed that three different particle sizes of OIDF have different retention and crystallinity. Among the three OIDFs, OIDF-10 exhibited the stronger WSC, OHC, CAC, and SCAC. The results after the feeding showed that the OIDF of three particle sizes could improve the elevation of blood lipids and the disturbance of gut microbiota caused by the high-fat diet. Therefore, this study demonstrated the functional significance of the three particle sizes of OIDF and provided a reference for its application in functional food processing, aiming at maintaining healthy blood lipid and intestinal flora levels.

Cis-bifenthrin inhibits cortisol and aldosterone biosynthesis in human adrenocortical H295R cells via cAMP signaling cascade.

Cis-bifenthrin (cis-BF) is a common-used pyrethroid insecticide frequently detected in environmental and biological matrices. Mounting evidence highlights the endocrine disrupting effects of cis-BF due to anti-estrogenic or anti-androgenic activity. However, little is known about the exposure effects of cis-BF on adrenal cortex function. In this study, effects of cis-BF on biosynthesis of adrenal steroids, as well as the potential mechanisms were investigated in human adrenocortical carcinoma (H295R) cells. Cis-BF decreased basal production levels of cortisol and aldosterone, as well as cAMP-induced production of cortisol. Both he basal and cAMP-stimulated transcriptional levels of several steroidogenic genes were significantly down-regulated by cis-BF. As an important rate-limiting enzyme in steroidogenesis, the protein level of StAR was prohibited by cis-BF on both basal and cAMP-induced conditions. Intracellular level of cAMP was significantly reduced by cis-BF. Overall, these data suggest that cis-BF may inhibit the biosynthesis of cortisol and aldosterone via disrupting cAMP signaling cascade.

Structural properties, antioxidant and immune activities of low molecular weight peptides from soybean dregs (Okara).

In this study, a method for preparing low molecular weight peptides (HPH-VAP) from okara using high-pressure homogenization assisted double enzymes was proposed. In order to explore its advantages, the effects of various methods on protein extraction rate and on the structure, antioxidant and immune properties of peptides were compared. The results showed that the protein extraction rate of this method was increased by 69% and 51% compared with other methods, and the structure only led to changes in the hydrogen bonds between peptide chains. HPH-VAP was screened out through functional characteristics, its structure was identified by HPLC-MS/MS, and further immunological activity analysis was carried out. The results showed that it promoted cell phagocytic ability, NO level and release of cytokines IL-6, IFN- γ, TNF-α. Therefore, this method is an effective and applicable method for industrial preparation of okara peptides, and has a positive effect on the reuse of okara resources.

Gas-solid phase flow synthesis of Cu-Co-1,3,5-benzenetricarboxylate for electrocatalytic oxygen evolution.

Using an environmentally friendly method to produce a stable and highly catalytically active electrocatalyst for the oxygen evolution reaction (OER) is becoming increasingly urgent. Herein, a novel bimetallic metal-organic framework (MOF), specifically a copper-cobalt 1, 3, 5-benzenetricarboxylate (Cu-Co-BTC) MOF, was successfully prepared by employing the gas-solid two-phase flow (GSF) synthetic technique. The as-prepared Cu-Co-BTC with its multiple active sites afforded a current density of 10 mA cm-2 at 239 mV for the OER in a 1 mol L-1 KOH solution, and showed a better electrocatalytic performance than did single-metal-containing Cu-BTC and Co-BTC materials. This work provides a new idea, one involving using novel gas-solid phase reactions for the preparation of electrocatalysts in large quantities.

A photofabricated honeycomb micropillar array for loss-free trapping of microfluidic droplets and application to digital PCR.

Droplet microfluidics is a promising platform for various biological and biomedical applications. Among which, droplet-based digital PCR (ddPCR) is one of the most challenging examples, with practical issues involving possible fusion/fission of droplets during PCR thermocycling and difficulties of indexing them for real-time monitoring. While spatially trapped droplet arrays may be helpful, they currently are either of low trapping density or suffer from high droplet loss. In this paper, we, for the first time, report a photofabricated honeycomb micropillar array (PHMA) for high-density and loss-free droplet trapping. By rationally designing high-aspect-ratio micropillars into a honeycomb configuration, droplets can be captured at a density of 160-250 droplets per mm2 and, more interestingly, without any loss. The PHMA device can be fabricated from several photocurable materials, with one gasproof photopolymer being optimally selected herein to enable the simple design to avoid sample evaporation and tedious surface modification, thereby making the fabrication very convenient. Moreover, by using a photocurable oil as a continuous phase, the trapped droplets can be further immobilized, and thus, become more stable even in PCR thermocycling. With these features, the proposed PHMA has shown promising potential in ddPCR, and is expected to find a wide range of applications in various biological and biomedical research.

Preparation and characterization of soybean insoluble dietary fiber and its prebiotic effect on dyslipidemia and hepatic steatosis in high fat-fed C57BL/6J mice.

The potential benefits of insoluble dietary fiber (IDF) in the regulation of lipid metabolism have been reported in large prospective cohort studies although the molecular regulatory mechanism is still unclear. Okara is a by-product obtained during soybean processing for soy milk and soybean curd (tofu), which is rarely utilized and can be a cheap potential dietary fiber (DF) resource. In this study, the structure and physicochemical properties of insoluble dietary fiber (SIDF) extracted from okara were characterized, and the prebiotic effects on fat metabolism were investigated in vivo. The results showed that the main monosaccharides of SIDF (90.50%) identified were galactose, arabinose, xylose, rhamnose and glucose. Scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), and X-ray diffraction (XRD) analyses suggested that SIDF had a loose and porous structure, polysaccharide functional groups, and a typical crystalline cellulose I structure. In addition, SIDF had ideal oil-adsorption capacity (OAC; 7.95 g g-1) and significantly improved cholesterol adsorption (11.14 mg g-1) at pH 7.0. In vivo, IDF supplementation reduced the serum lipid levels and inhibited hepatic fat accumulation. Additionally, SIDF administration improved hepatic steatosis by stimulating lipolysis via upregulation of PPARα, CYP4a10 and CPT1a. This is the first systematic study on the composition, structure, physicochemical properties, adsorption function and biological effects of SIDF. The above results show that SIDF could be used as an ideal functional ingredient in food processing as well as play a positive role in improving the added value of okara and promoting its comprehensive utilization.

Composite Elastomer-Enabled Rapid Photofabrication of Microfluidic Devices.

Microfluidics, as an emerging technology, is highly dependent on the evolution of device materials and fabrication techniques. While replica molding of polydimethylsiloxane and hot embossing/injection molding of thermoplastics are most popular, they are either hard to scale up or inappropriate for laboratory-scale prototyping. Recently, photocurable resins, as a huge class of materials, have attracted extensive interest. However, very few of them can now be used in device fabrication due to the challenge in machining these materials. In response, we herein propose a novel concept of composite elastomers, which can covalently link with and consequently offer a flexible support to photocured thin films. This effect would allow most photocurable resins to be used in microfluidic device fabrication, greatly enriching the material choices for diverse applications. Moreover, the whole fabrication process becomes very simple and rapid, with an impressive throughput of at least hundreds of replicas per day. With these features, it is reasonably expected that the composite elastomer-enabled rapid photofabrication method will be very competent for laboratory prototyping, providing not only the ease of fabrication but also a possibility to select the materials specifically for ultimate applications and promising potential for volume production without the redevelopment process. These may offer a good opportunity to narrow the current gap between academic research and industrial practice.

An Intelligent Self-Service Vending System for Smart Retail.

The traditional weighing and selling process of non-barcode items requires manual service, which not only consumes manpower and material resources but is also more prone to errors or omissions of data. This paper proposes an intelligent self-service vending system embedded with a single camera to detect multiple products in real-time performance without any labels, and the system realizes the integration of weighing, identification, and online settlement in the process of non-barcode items. The system includes a self-service vending device and a multi-device data management platform. The flexible configuration of the structure gives the system the possibility of identifying fruits from multiple angles. The height of the system can be adjusted to provide self-service for people of different heights; then, deep learning skill is applied implementing product detection, and real-time multi-object detection technology is utilized in the image-based checkout system. In addition, on the multi-device data management platform, the information docking between embedded devices, WeChat applets, Alipay, and the database platform can be implemented. We conducted experiments to verify the accuracy of the measurement. The experimental results demonstrate that the correlation coefficient R2 between the measured value of the weight and the actual value is 0.99, and the accuracy of non-barcode item prediction is 93.73%. In Yangpu District, Shanghai, a comprehensive application scenario experiment was also conducted, proving that our system can effectively deal with the challenges of various sales situations.

Prebiotic Impacts of Soybean Residue (Okara) on Eubiosis/Dysbiosis Condition of the Gut and the Possible Effects on Liver and Kidney Functions.

Okara is a white-yellow fibrous residue consisting of the insoluble fraction of the soybean seeds remaining after extraction of the aqueous fraction during the production of tofu and soymilk, and is generally considered a waste product. It is packed with a significant number of proteins, isoflavones, soluble and insoluble fibers, soyasaponins, and other mineral elements, which are all attributed with health merits. With the increasing production of soy beverages, huge quantities of this by-product are produced annually, which poses significant disposal problems and financial issues for producers. Extensive studies have been done on the biological activities, nutritional values, and chemical composition of okara as well as its potential utilization. Owing to its peculiar rich fiber composition and low cost of production, okara might be potentially useful in the food industry as a functional ingredient or good raw material and could be used as a dietary supplement to prevent varied ailments such as prevention of diabetes, hyperlipidemia, obesity, as well as to stimulate the growth of intestinal microbes and production of microbe-derived metabolites (xenometabolites), since gut dysbiosis (imbalanced microbiota) has been implicated in the progression of several complex diseases. This review seeks to compile scientific research on the bioactive compounds in soybean residue (okara) and discuss the possible prebiotic impact of this fiber-rich residue as a functional diet on eubiosis/dysbiosis condition of the gut, as well as the consequential influence on liver and kidney functions, to facilitate a detailed knowledge base for further exploration, implementation, and development.

A comparison of removal efficiencies of conventional drinking water treatment and advanced treatment equipped with ozone-biological activated carbon process.

ABSTRACTUsing raw water from a shallow water supply reservoir located in the lower Yangtze River region, the removal efficiencies of conventional treatment on dissolved organic matter (DOM) and disinfection by-products formation potential (DBPFP) were compared with an advanced treatment that equipped with ozone-biological activated carbon (O3-BAC) process. The results showed that the advanced treatment was more efficient than the conventional treatment at removing dissolved organic carbon (DOC; 40-67% removal), UV254 (61-81% removal), the trihalomethane formation potential (THMFP; 37-70% removal) and the haloacetic acid formation potential (HAAFP; 35-89% removal). The sand filter in the conventional treatment process was identified as the main contributor to decreasing DOC, UV254 and DBPFP. The O3-BAC in advanced treatment was found to decrease THMFP and HAAFP, with removal rates of 17-40% and 22-59%, respectively. To improve the water quality of effluents, advanced treatment with O3-BAC should be used to treat raw water from the shallow water supply reservoir in lower Yangtze River. However, the increased DBPFP yield, which is proportional to the potential health risks, should not be ignored.

Aligner-Mediated Cleavage-Based Isothermal Amplification for SARS-CoV-2 RNA Detection.

Rapid detection of SARS-CoV-2 RNA is critical for reducing the global transmission of COVID-19. Here, we report a simple and versatile assay for detection of SARS-CoV-2 RNA based on aligner-mediated cleavage-based strand displacement amplification (AMC-SDA). The entire amplification procedure takes less than 25 min without professional instruments or requirement of specific target sequences and can reach a limit of detection of attomolar RNA concentration. Using pseudovirus as mimicry of clinical SARS-CoV-2 positive samples, we achieved a diagnostic accuracy of 100% in 10 simulated samples (five positive and five negative). We anticipate that our method will provide a universal platform for rapid and accurate detection of emerging infectious diseases.

Efficacy and safety of micro-implant anchorage in Angle class II malocclusion orthodontic treatment: A protocol for systematic review and meta-analysis.

BACKGROUND: Angle class II malocclusion is clinically complex and common malocclusion type, which affects beauty. Conventional treatment has the disadvantages of long course of treatment, high cost, easy recurrence and limited curative effect. Clinical practice shows that micro-implant anchorage has certain advantages in the treatment of Angle II malocclusion, but lacks the evidence of evidence-based medicine. This study systematically evaluates the efficacy and safety of micro-implant anchorage in the treatment of Angle class II malocclusion. METHODS: A systematic search was performed by retrieving on English databases (PubMed, Embase, Web of Science, and the Cochrane Library) and Chinese databases (CNKI, Wanfang, Weipu [VIP], CBM). Besides, manually search for Google and Baidu academic of micro-implant anchorage in the treatment of Angle class II malocclusion in randomized controlled clinical research. The retrieval time limit was from the establishment of the database to September 2020. Two researchers independently extracted and evaluated the quality of the data in the included study. A meta-analysis was performed using RevMan5.3 software. RESULTS: In this study, the efficacy and safety of micro-implant anchorage against Angle class II malocclusion were evaluated by SNA, BNA, ANB, NLA°, Adverse reaction. CONCLUSIONS: This study will provide reliable evidence-based evidence for the clinical application of micro-implant anchorage in the treatment of Angle class II malocclusion. ETHICS AND DISSEMINATION: Private information from individuals will not be published. This systematic review also does not involve endangering participant rights. Ethical approval was not required. The results may be published in a peer-reviewed journal or disseminated at relevant conferences.OSF Registration number: DOI 10.17605/OSF.IO/UPBR8.

Rapid In Situ Photoimmobilization of a Planar Droplet Array for Digital PCR.

Digital PCR (dPCR) is a powerful technique capable of absolute quantification of nucleic acids with good accuracy. Droplet-based dPCR (ddPCR), among others, is one of the most important dPCR techniques. However, the surface tension-controlled droplets may suffer from fusion/fission due to the vigorous temperature change in PCR thermal cycling. Besides, the free movement of droplets makes them unsuitable for real-time fluorescence monitoring. In this paper, we first developed a photoimmobilized planar droplet array (PIPDA) by using a photocurable polyurethane as the continuous oil phase. It is found that uniform water-in-oil droplets of various sizes can be readily generated, and more importantly, the oil phase can be rapidly solidified in just a few seconds upon exposure to UV irradiation. This process will leave the droplets immobilized in the accommodation chamber as a stable planar array and, thus, effectively prevent the movement, coalescence, and breakup of droplets. In addition, a novel multilayered chip design has been proposed, which can thoroughly overcome the evaporation issue that commonly exists in polydimethylsiloxane (PDMS)-based dPCR chips. With these two innovations, the ddPCR experiment could be performed in a robust manner, and shows a promising potential in the development of real-time ddPCR technique. These features may therefore enable the wide application of PIPDA-based ddPCR in various fields.

Aligner-mediated cleavage-triggered exponential amplification for sensitive detection of nucleic acids.

Exponential amplification reaction (EXPAR), as a simple and high sensitive method, holds great promise in nucleic acids detection. One major challenge in EXPAR is the generation of trigger DNA with a definite 3'-end, which now relies on fingerprinting technology. However, the requirement of different endonucleases for varying target sequences and two head-to-head recognition sites in double stranded DNA, as well as the confinement of trigger DNA's 3'-end to be near/within the recognition site, usually subject EXPAR to compromised universality and/or repeated matching of reaction conditions. Herein, we report a simple and universal method for high sensitive detection of nucleic acids, termed aligner-mediated cleavage-triggered exponential amplification (AMCEA). The aligner-mediated cleavage (AMC) needs only one nicking endonuclease and can make a break at any site of choice in a programmable way. Thus, the 3'-end of target DNA can be easily redefined as required, a key step for initiating the amplification reaction. This capability endows the proposed AMCEA with excellent universality and simplicity. Moreover, it is sensitive and specific, with a detection limit at amol level, a broad dynamic range of 5～6 orders of magnitude and the ability to distinguish single nucleotide mutation. Experiments performed with human serum indicate that AMCEA is compatible with the complex biological sample, and thus has the potentials for practical applications.

Aligner-mediated cleavage of nucleic acids and its application to isothermal exponential amplification.

We herein describe a simple and versatile approach to use conventional nicking endonuclease (NEase) for programmable sequence-specific cleavage of DNA, termed aligner-mediated cleavage (AMC), and its application to DNA isothermal exponential amplification (AMC-based strand displacement amplification, AMC-SDA). AMC uses a hairpin-shaped DNA aligner (DA) that contains a recognition site in its stem and two side arms complementary to target DNA. Thus, it enables the loading of an NEase on DA's stem, localization to a specific locus through hybridization of the side arms with target DNA, and cleavage thereof. By using just one NEase, it is easy to make a break at any specific locus and tune the cleavage site to the single-nucleotide scale. This capability also endows the proposed AMC-SDA with excellent universality, since the cleavage of target DNA, followed by a polymerase-catalyzed extension along a particular primer as a key step for initiating SDA, no longer relies on any special sequence. Moreover, this manner of initiation facilitates the adoption of 3'-terminated primers, thus making AMC-SDA highly sensitive and highly specific, as well as simple primer design.

Prevalence and molecular characterization of fluoroquinolone resistance in Escherichia coli isolates from dairy cattle with endometritis in China.

Fluoroquinolones are frequently used to treat infectious disease that is caused by Escherichia coli in dairy cattle. However, fluoroquinolone resistance occurs and is due either to chromosomal mutations in the bacterial topoisomerase genes and/or to plasmid-mediated resistance genes. The purpose of this study was to determine the prevalence and molecular characteristics of fluoroquinolone resistance determinants in E. coli strains (n=148) isolated from dairy cattle with bovine endometritis in Inner Mongolia (China). Analysis of the mutations in the quinolone resistance-determining regions of resistant E. coli isolates confirmed previously reported substitutions in the GyrA and ParE. However, we identified additional substitutions in the ParC and GyrB that have not been reported earlier. No plasmid-mediated quinolone resistance genes in any of the isolates were found. The number of point mutations found per isolate correlated with an increase in the minimum inhibitory concentration of ciprofloxacin. Overall, 45.5% of the isolates were positive for the class I integrase gene along with four gene cassettes that were responsible for resistance to trimethoprim (dfr1 and dfrA17) and aminoglycosides (aadA1 and aadA5), respectively. The prevalence of extended-spectrum ß-lactamases (ESBLs) was 100%, and the blaTEM gene was predominant in all of the isolates. In conclusion, our results identify the mechanism of quinolone resistance for the first time and reveal the prevalence of integron and ESBLs in E. coli isolates from dairy cattle with bovine endometritis in China after 20 years of quinolone usage in cattle.

Soft nanomaterial-based targeting polymersomes for near-infrared fluorescence multispectral in vivo imaging.

We report here the soft nanomaterial-based targeting polymersomes for near-infrared (NIR) fluorescence imaging to carry out in vivo tumor detection. Two polymersome-based NIR fluorescent probes were prepared through the self-assembly of amphiphilic block copolymers, poly(butadiene-b-ethylene oxide) (PEO-b-PBD). Each of them was encapsulated with distinct hydrophobic near-infrared dyes (DiD and DiR) and modified with different targeting ligands (anti-CEA antibody and anti-EGFR antibody), respectively. After simultaneous injection of these two probes into the tumor-bearing mice via tail vein, multispectral near-infrared fluorescence images were obtained. The results indicate that both probes are successfully directed to the tumor foci, where two distinguishable fluorescent signals were detected through the unmixed fluorescence images. By taking advantage of two targeting polymersome-based probes with distinct fluorescent features, the proposed multispectral near-infrared fluorescence imaging method can greatly improve the specificity and accuracy for in vivo tumor detection.