Development of a reverse transcription loop-mediated isothermal amplification based clustered regularly interspaced short palindromic repeats Cas12a assay for duck Tembusu virus.

Duck Tembusu virus (DTMUV) is an emerging pathogen that poses a serious threat to the duck industry in China. Currently, polymerase chain reaction (PCR), quantitative PCR (qPCR) and reverse transcription loop-mediated isothermal amplification (RT-LAMP) are commonly used for DTMUV detection. However, these methods require complex steps and special equipment and easily cause false-positive results. Therefore, we urgently need to establish a simple, sensitive and specific method for the clinical field detection of DTMUV. In this study, we developed an RT-LAMP-based CRISPR-Cas12a assay targeting the C gene to detect DTMUV with a limited detection of 3 copies/µL. This assay was specific for DTMUV without cross-reaction with other common avian viruses and only required some simple pieces of equipment, such as a thermostat water bath and blue/UV light transilluminator. Furthermore, this assay showed 100% positive predictive agreement (PPA) and negative predictive agreement (NPA) relative to SYBR Green qPCR for DTMUV detection in 32 cloacal swabs and 22 tissue samples, supporting its application for clinical field detection.

Supply chain finance and innovation efficiency: An empirical analysis based on manufacturing SMEs.

This paper firstly demonstrates the positive and negative effects of supply chain finance on the innovation efficiency of China's small and medium-sized enterprises (SMEs) in the manufacturing industry from the theoretical point of view. Based on the data of 267 manufacturing companies in China Growth Enterprise Market from 2015 to 2019, the DEA-SBM method was used to measure the comprehensive innovation efficiency of different companies, and it was further decomposed into technological innovation efficiency and organizational innovation efficiency. Afterwards, it conducts an empirical analysis through the double fixed effect model, and explores the difference in the impact of supply chain finance on innovation efficiency in enterprises with different industries and different property rights. The results show that supply chain financial services have a strong positive impact on the comprehensive innovation efficiency, technological innovation efficiency and organizational innovation efficiency of manufacturing SMEs. Further, supply chain finance has the most significant improvement on the technological innovation efficiency of the sample of private traditional enterprises, but it has a significant inhibitory effect on the organizational innovation efficiency of the sample of state-owned high-tech enterprises. Therefore, this paper suggests that the development of supply chain financial services should increase support for traditional manufacturing industries; appropriately tilt resources to private enterprises; improve relevant supply chain financial laws and regulations, establish and improve corresponding institutional arrangements, and encourage state-owned enterprises to participate in market competition.

Metallic foreign bodies ingestion by schizophrenic patient: a case report.

The ingestion of foreign objects is a widespread health issue, with a higher occurrence in adults with psychosis. Case Presentation: The authors present the case of a 39-year-old man who arrived at the hospital with symptoms of abdominal distension and occasional black stools for a week. The patient was known to have schizophrenia but had not received regular hospital follow-up or treatment for the past 5 years. He had a history of exogenous stimulation, which led him to surreptitiously swallow metallic objects. Upon physical examination, he displayed abdominal distension and mild tenderness in the upper abdomen. Radiographs revealed multiple foreign objects in his stomach, leading to the decision for laparotomy, gastric opening, and removal of the foreign objects under general anesthesia. Clinical Discussion: Mental illnesses, including schizophrenia, bipolar disorder, major depression, and multiple substance abuse, are recognized as being significant risk factors for ingesting foreign bodies. In such cases, it is crucial to intervene quickly. For patients presenting with psychiatric symptoms, the involvement of family caregivers is of even greater importance than endoscopic or surgical treatments. Conclusion: Foreign body ingestion is more prevalent in individuals with psychosis, highlighting the importance of ongoing care and follow-up for patients with mental illness.

Portable paper-based electrochemiluminescence test incorporating lateral-flow immunosensors for detection of interferon-γ levels.

Tuberculosis (TB) poses a serious threat to human health and social development. Accurate diagnosis of mycobacterium tuberculosis infection plays a critical role in the prevention and treatment of tuberculosis. Interferon-γ (INF-γ) release assay (IGRA) is currently the only quantitative tuberculosis infection diagnosis method. An accurate, fast, and easily handled INF-γ detection method is the key to obtaining accurate results. Herein, we report a novel paper-based electrochemiluminescence (ECL) method based on lateral flow immunosensors that combines the easy handling characteristics of immunochromatography and the high sensitivity of electrochemiluminescence to detect IFN-γ. To our knowledge this is the first INF-γ detection method that combines immunochromatography with electrochemiluminescence. The paper-based ECL-LFI test consists of a sample pad, conjugation pad (with binding antibody IFN-γ-Ab1 conjugated with ruthenium tripyridine), detection pad (with capture antibody IFN-γ-Ab2 immobilized on nanospheres), absorbent pad, and electrode for signal activation. The ECL signal is obtained by cyclic voltammetry scanning at a speed of 0.1 V/s in the detection area of the paper-based ECL-LFI test. In our experiments, the paper-based ECL-LFI test exhibited a minimum detection limit of 2.57 pg/mL within 12 min, and a broad detection range of 2.57-5,000 pg/mL, with repeatability of 8.10% and stability of 4.97%. With the advantage of high accuracy and sensitivity, easy handling, and low user training requirements, this ECL-LFI test might be used as point-of-care testing (POCT) in the IGRA for tuberculosis diagnosis.

[Temporal and Spatial Distribution Characteristics and Difference Analysis of Chromophoric Dissolved Organic Matter in Sediment Interstitial Water from Gangnan Reservoir].

The chromophoric dissolved organic matter (CDOM), the main component of dissolved organic matter, affects the morphological characteristics, migration, and conversion of pollutants in water. Based on UV-vis spectra and excitation emission matrix spectroscopy (EEMs) combined with the parallel factor analysis (PARAFAC), the spatial distribution and spectral characteristics were investigated and source analysis of CDOM was performed. Thus, the spatiotemporal differences in the CDOM in Gangnan Reservoir were analyzed. Results showed that a254, a260, a280, and a355 exhibited significant seasonal differences in Gangnan Reservoir, and the order of CDOM concentrations was summer > spring > autumn > winter. There are significant seasonal differences in the E2/E3, E3/E4, E4/E6, and SR of interstitial water CDOM. The concentrations of E2/E3, E3/E4, E4/E6, and SR were high in winter and low in summer. E2/E3 and E3/E4 in autumn and winter were significantly higher than those in spring and summer, and the E3/E4 in autumn and winter was greater than 3.5, which indicates that the CDOM of the autumn and winter sediments has a smaller molecular weight and a lower degree of humification. Protein-like substances (C1), short-wave fulvic acid (C2), and degraded humic substances (C3) were identified by the PARAFAC model, and there was a significant positive correlation among the three fluorescent components (P<0.001). The total fluorescence intensity of CDOM and the fluorescence intensity of each fluorescent component show significant seasonal differences. The total fluorescence intensity and the fluorescence intensity of each component show the highest levels in spring, followed by autumn and winter, and the lowest levels in summer. The proportion of each fluorescent component in autumn and winter and that of each fluorescent component in spring and summer showed no significant difference. There was a significant difference in the proportion of each fluorescent component between autumn/winter and spring/summer. The BIX and FI of CDOM for autumn and winter were higher than those for spring and summer, indicating that the autogenous source of CDOM in autumn and winter is stronger than that in spring and summer, which was consistent with the result of HIX. PCA and Adonis analysis showed that the spectral characteristics of CDOM exhibited obvious seasonal differences (P<0.001). Moreover, the C1, C2, and C3 and water quality parameters (NH4+, NO3-, NO2-, TDN, and TDP) exhibited significant correlation based on linear regression. The results could provide technical support for the control of organic carbon pollution sources and water quality management in Gangnan Reservoir.

[Spatial Distribution Characteristics and Driving Factors of Aerobic Denitrification Bacterial Community Structure from Baiyangdian Lake in Xiong'an New Area During the Winter Freezing Period].

To isolate the aerobic denitrification bacteria suitable for water quality in the low-temperature period of Baiyangdian Lake, a water quality investigation and bioinformatics analysis of the aerobic denitrification bacterial community were carried out using a MiSeq high-throughput sequencing technique based on napA. Moreover, α-diversity, ß-diversity, and network analyses were also carried out. The results showed significant differences in the water quality of different sampling sites in Baiyangdian Lake, and the estuary area exhibited the highest nitrogen concentration. α-diversity exhibited significant differences (P<0.05), and the abundance and diversity of ZZD and BH were the lowest. The operational taxonomic units of the water body mainly belonged to Proteobacteria (α-Proteobacteria, ß-Proteobacteria, and γ-Proteobacteria). Meanwhile, Venn diagram analysis indicated the community of aerobic denitrification bacteria exhibited significant differences, and variance inflation factor and redundancy analysis showed that temperature, dissolved oxygen, ammonia, nitrate, dissolved total phosphorus, and redox potential were the main environmental factors. Network analysis showed that symbiotic relationships accounted for a major proportion of the microbial network. Mantel test analysis shows that temperature, redox potential, nitrate, ammonia, dissolved total phosphorus, and iron and manganese are the key factors affecting the evolution of modular community structure. From all the results, the MiSeq high-throughput sequencing technique based on the napA gene was an effective tool to explore the changes of aerobic denitrification bacterial community structure, which could supply a reference to isolate the "directional-accurate-efficient" aerobic denitrification bacterial agent in the future.

Complete genome sequence of an IMP-8, CTX-M-14, CTX-M-3 and QnrS1 co-producing Enterobacter asburiae isolate from a patient with wound infection.

OBJECTIVES: The aim of this study was to investigate the characteristics and complete genome sequence of an IMP-8, CTX-M-14, CTX-M-3 and QnrS1 co-producing multidrug-resistant Enterobacter asburiae isolate (EN3600) from a patient with wound infection. METHODS: Species identification was confirmed by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS). Carbapenemase genes were identified by PCR and Sanger sequencing. The complete genome sequence of E. asburiae EN3600 was obtained using a PacBio RS II platform. Genome annotation was done by Rapid Annotation using Subsystem Technology (RAST) server. Acquired antimicrobial resistance genes (ARGs) and plasmid replicons were detected using ResFinder 2.1 and PlasmidFinder 1.3, respectively. RESULTS: The genome of E. asburiae EN3600 consists of a 4.8-Mbp chromosome and five plasmids. The annotated genome contains various ARGs conferring resistance to aminoglycosides, ß-lactams, fluoroquinolones, fosfomycin, macrolides, phenicols, rifampicin and sulfonamides. In addition, plasmids of incompatibility (Inc) groups IncHI2A, IncFIB(pECLA), IncFIB(pQil) and IncP1 were identified. The genes blaIMP-8, blaCTX-M-14 and blaCTX-M-3 were located on different plasmids. The blaIMP-8 gene was carried by an 86-kb IncFIB(pQil) plasmid. The blaCTX-M-3 and qnrS1 genes were co-harboured by an IncP1 plasmid. In addition, blaCTX-M-14 was associated with blaTEM-1B, blaOXA-1, catB3 and sul1 genes in a 116-kb non-typeable plasmid. CONCLUSION: To our knowledge, this is the first complete genome sequence of an E. asburiae isolate co-producing IMP-8, CTX-M-14, CTX-M-3 and QnrS1. This genome may facilitate the understanding of the resistome, pathogenesis and genomic features of Enterobacter cloacae complex (ECC) and will provide valuable information for accurate identification of ECC.

Screening key lncRNAs for human rectal adenocarcinoma based on lncRNA-mRNA functional synergistic network.

BACKGROUND: Rectal adenocarcinoma (READ) is one of the deadliest malignancies, and the molecular mechanisms underlying the initiation and development of READ remain largely unknown. In this study, we aimed to find key long noncoding RNAs (lncRNAs) and mRNAs in READ by RNA sequencing. METHODS: RNA sequencing was performed to identify differentially expressed mRNAs (DEmRNAs) and lncRNAs (DElncRNAs) between READ and normal tissue. READ-specific protein-protein interaction (PPI), DElncRNA-DEmRNA coexpression, and DElncRNA-nearby DEmRNA interaction networks were constructed. DEmRNAs and DEmRNAs coexpressed with DElncRNAs were functionally annotated. RESULTS: A total of 2113 DEmRNAs and 150 DElncRNAs between READ and normal tissue were identified. The PPI network identified several hub proteins, including CDK1, AURKB, CDC6, FOXQ1, NUF2, and TOP2A. The DElncRNA-DEmRNA coexpression and DElncRNA-nearby DEmRNA interaction networks identified some hub lncRNAs, including CCAT1, LOC105374879, GAS5, and B3GALT5-AS1. The colorectal cancer pathway, the intestinal immune network for IgA production and the p53 signaling pathway were three pathways significantly enriched in DEmRNAs and DEmRNAs coexpressed with DElncRNAs. MSH6 coexpressed with two DElncRNAs (LOC105374879 and CASC15) and BCL2 coexpressed with B3GALT5-AS1 were significantly enriched in the colorectal cancer signaling pathway. TNFRSF17 coexpressed with B3GALT5-AS1 was enriched in the intestinal immune network for IgA production. CCNB2 coexpressed with LOC105374879 was enriched in the p53 signaling pathway. CONCLUSION: A total of four DEmRNAs (MSH6, BCL2, TNFRSF17, and CCNB2) and three DElncRNAs (LOC105374879, CASC15, and B3GALT5-AS1) may be involved in the pathogenesis of READ; this data may contribute to understanding the mechanisms of READ and the development of therapeutic strategies for READ.

The KLF14 Transcription Factor Regulates Glycolysis by Downregulating LDHB in Colorectal Cancer.

The Krüppel-like transcription factor 14 (KLF14) is a critical regulator of a wide array of biological processes. However, the role of KLF14 in colorectal cancer (CRC) isn't fully investigated. This study aimed to explore the clinicopathological significance and potential role of KLF14 in the carcinogenesis and progression of CRC. A tissue microarray consisting of 185 samples from stage I-III CRC patients was adopted to analyze the correlation between KLF14 expression and clinicopathological parameters, as well as overall survival (OS) and disease-free survival (DFS). The underlying mechanisms of altered KLF14 expression on glycolysis were studied using in vitro and patients' samples. The results showed that KLF14 expression was downregulated in CRC than their normal controls. Low KLF14 expression correlated with advanced T stage (P< 0.001) and N stage (P= 0.040), and larger tumor size (P= 0.008). Lost KLF14 expression implied shorter OS and DFS after colectomy in both univariate and multivariate survival analysis (P<0.05). Experimentally, restore KLF14 expression significantly decreased the rate of glycolysis both in vitro and in patients' sample. Mechanically, KLF14 regulated glycolysis by downregulating glycolytic enzyme LDHB. Collectively, KLF14 is a novel prognostic biomarker for survival in CRC, and downregulation of KLF14 in CRC prompts glycolysis by target LDHB. Hence, KLF14 could constitute potential prognostic predictors and therapeutic targets for CRC.

NOTCH4 Is a Novel Prognostic Marker that Correlates with Colorectal Cancer Progression and Prognosis.

Notch family plays vital role in carcinogenesis and progression of various cancer, however, its clinical significance and prognostic value in colorectal cancer isn't fully investigated. In present study, we first investigated the NOTCH4 expression in The Cancer Genome Atlas (TCGA) (n=361) and GSE39582 (n=474) database and then validated with our own database (n=248). The transcriptional and protein levels of NOTCH4 were evaluated by RT-PCR and immunohistochemistry study, respectively. Univariate and multivariate survival analyses were performed to explore the relationship between various prognostic factors and survival outcomes. In the univariate analysis, NOTCH3 and NOTCH4 were significantly correlated with prognosis in TCGA and GSE39582 database, respectively (P<0.05). For NOTCH3 has been studied in CRC, we chosen NOTCH4 for further study. NOTCH4 mRNA was higher in liver metastases than their primary colorectal cancer or normal mucosa. Increased NOTCH4 levels significantly correlated with advanced N stage (P= 0.002), M stage (P= 0.002), lymphovascular invasion (P= 0.026), and CEA status (P= 0.030). Patients with high NOTCH4 expression had shorter 5-year disease-free survival (DFS) (HR 6.809; 95% CI 3.334-13.904; P< 0.001) and overall survival (OS) (HR 6.476; 95% CI 3.307-12.689; P<0.001) than those with low NOTCH4 expression. Multivariate survival analysis demonstrated that NOTCH4 was an independent prognostic biomarker for both DFS (HR 7.848; 95% CI 3.777-16.308; P<0.001) and OS (HR 5.323; 95% CI 2.668-10.623; P<0.001).Collectively, NOTCH4 may play critical role in colorectal cancer progression and could serve as a novel biomarker to predict survival after colectomy.

Colloid properties of hydrophobic modified alginate: Surface tension, ζ-potential, viscosity and emulsification.

Micelle properties of hydrophobic modified alginate (HM-alginate) in various dispersion media have been studied by surface tension, ζ-potential, and viscosity measurements. Effect of salt on micelle properties showed that the presence of counter ion weakened the repulsive interaction between surfactant ions, decreased the critical micelle concentration (CMC) value of the HM-alginate, reduced the effective volume dimensions of HM-alginate and hence viscosity, which coincide with the corresponding ζ-potential values. Soy oil-in-water emulsions, stabilized solely by HM-alginate, were produced in high speed homogenization conditions and their stability properties were studied by visual inspection, optical microscopy and droplet size measurements. The results showed that emulsions (oil-water ratio was 1:7) containing 15mg/mL HM-alginate presented better stability during 15days storage, which stating clearly that HM-alginate is an effective emulsifier to stabilize oil-in-water emulsions. The herein presented homogeneous method for preparation of emulsion has the potential to be used in food industry.

[Prognostic value of lymph node ratio in survival of patients with stage III rectal cancer].

OBJECTIVE: To assess the prognostic value of lymph node ratio (LNR) in patients with stage III rectal cancer after curative resection. METHODS: A retrospective review of the clinicopathological data was performed in 161 patients with stage III rectal cancer who received curative surgical excision in our hospital from June 2005 to June 2010. The variables including LNR, age, gender, T stage, N stage, total number of dissected lymph nodes, number of metastatic lymph nodes, and positive rate of lymph node metastasis were studied through univariate and multivariate analyses, and the survival analysis was performed using Kaplan-Meier method and Log rank test. RESULTS: Multivariate analysis revealed that LNR, but not number of positive nodes or number of harvested lymph nodes, had independent prognostic value for overall survival and disease-free survival for patients with stage III rectal cancer. The overall survival in the LNR < 0.43 and LNR ≥ 0.43 groups was 75.8% and 41.3%, respectively (P < 0.01), while the disease-free survival was 68.8% and 40.3%, respectively (P = 0.001). CONCLUSIONS: The LNR is an independent prognostic factor for survival of patients with stage III rectal cancer, and is more efficient than the number of positive nodes and total number of dissected lymph nodes in the survival prediction.