One hundred and thirty-six bla OXA-51-negative strains were identified from 1,067 Acinetobacter calcoaceticus-A. baumannii complex (ACB complex) isolates, which were collected during October 2010 to March 2013 from 15 general hospitals in 10 cities throughout Zhejiang Province, China. Seven of the 136 bla OXA-51-negative ACB complex isolates were New Delhi metallo-β-lactamase-1 (NDM-1)-positive, among which three were identified as A. nosocomialis and four were identified as A. pittii strains using 16S-23S rRNA gene intergenic spacer (ITS) sequencing and partial RNA polymerase β-subunit (rpoB) sequencing. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) analysis showed that the seven NDM-positive isolates belonged to three clonal strains with three novel sequence types (STs). Polymerase chain reaction (PCR) assays and DNA sequence analysis of the carbapenemase and other β-lactamase genes indicated that all the isolates harbored the bla NDM-1 gene, and that only one strain of A. nosocomialis isolates harbored both bla NDM-1 and bla OXA-23. All of them were positive for bla ADC, from which three novel bla ADC genes (designated as bla ADC-69, bla ADC-70, and bla ADC-71) were detected for the first time. The presence of ISAba125 upstream of bla NDM-1 was identified through genetic environment analysis. Carbapenem resistance can be transferred from A. nosocomialis and A. pittii to Escherichia coli EC600 by the conjugation experiment. Plasmid analysis, DNA hybridization, and extraction experiments indicated that bla NDM-1 was located on a plasmid of approximately 50 kb. In conclusion, we characterized the dissemination of NDM-1-positive A. pittii strains in Zhejiang Province, China, and reported the NDM-producing A. nosocomialis for the first time.

Deflection measurement is the research focus of health monitoring for bridges during the operation period. This study develops a contactless measurement technique to monitor the bridge deflection, leveraging visual information from a team of unmanned aerial vehicles (UAVs). On the basis of the collinearity of the laser spots projected on the plane by the coplanar laser indicator, we can eliminate the motion of UAV, and calculate the vertical displacement of the position to be measured relative to the bridge pier. In the proposed method, the center of the laser spot is extracted through a method based on deep learning, and an algorithm based on scale-invariant features registration was developed to track the feature points of the bridge in the image sequence. According to the algorithm, we demonstrate the accuracy and feasibility of our approach through simulation and simulated bridge experiments. The result shows that the root mean squared error (RMSE) of measurement through our technique is less than 0.5 mm in the laboratory conditions. In addition, the limits and scalability of the presented method have been explored through a field experiment.

We investigated the distribution of resistance genes and the clonal relationships among carbapenem-resistant Acinetobacter baumannii isolates from the intensive care unit wards of two hospitals in Guangzhou, China. From 2012 to 2013, 57 A. baumannii isolates were obtained from blood cultures from two hospitals in Guangzhou. The antibiotic resistance profiles were determined by using the Vitek2 system and Etest strips. PCR was used to detect the genes encoding OXA-type carbapenemases and metallo-β-lactamases and the presence of ISAba1 upstream of the bla(OXA-51-like) gene and the bla(OXA-23-like) gene. Multilocus sequence typing (MLST) and sequence-based typing of bla(OXA-51-like) genes (SBT-bla(OXA-51-like )genes) were performed to analyze the genetic relationship of the isolates. Among the 57 isolates, 46 were carbapenem-resistant A. baumannii (CRAB) isolates. The bla(OXA-51-like) gene was identified in all 57 isolates, while the bla(OXA-23-like) gene was present in all 46 CRAB isolates. The MLST analysis grouped the A. baumannii isolates into five existing sequence types (STs) and five new STs. Fifty-two isolates belonged to the worldwide spread of clonal complex 92 (CC92), among which ST195 and ST365 were the most common STs. The MLST data and SBT-bla(OXA-51-like) genes showed that all isolates harboring the major bla(OXA-51-like) alleles, such as bla(OXA-66), belonged to CC92.

Interleukin-25 (IL17E/IL25) plays a critical role in colitis and intestinal homeostasis. However, the expression and biological role of IL25 in colorectal cancer is not properly understood. In this study, we show that IL25 is mainly expressed by cancer stem cells in the colorectal cancer microenvironment. Genetic deletion of IL25 inhibited tumor formation and growth and prolonged survival in AOM/DSS-treated mice. IL25 stimulated cancer organoid and cancer cells sphere formation and prevented the tumor from chemotherapy-induced apoptosis. Mechanistically, IL25 upregulated stem cell genes LGR5, CD133, and ABC transporters via activating the Hedgehog signaling pathway. IL25 inhibited phosphorylation of AMPK and promoted GLI1 accumulation to maintain cancer stem cells. Moreover, IL25 expression was associated with poor survival in patients with metastatic colorectal cancer. Taken together, our work reveals an immune-associated mechanism that intrinsically confers cancer cell stemness properties. Our results first demonstrated that IL25, as a new potent endogenous Hedgehog pathway agonist, could be an important prognostic factor and therapeutic target for CRC.

N6-methyladenosine (m6A) is the most prevalent RNA modification in eukaryotic mRNAs. Currently available detection methods for locus-specific m6A marks rely on RT-qPCR, radioactive methods, or high-throughput sequencing. Here, we develop a non-qPCR, ultrasensitive, isothermal, and naked-eye visible method for m6A detection based on rolling circle amplification (RCA) and loop-mediated isothermal amplification (LAMP), named m6A-Rol-LAMP, to verify putative m6A sites in transcripts obtained from the high-throughput data. When padlock probes hybridize to the potential m6A sites on targets, they are converted to circular form by DNA ligase in the absence of m6A modification, while m6A modification hinders the sealing of padlock probes. Subsequently, Bst DNA polymerase-mediated RCA and LAMP allow the amplification of the circular padlock probe to achieve the locus-specific detection of m6A. Following optimization and validation, m6A-Rol-LAMP can ultra-sensitively and quantitatively determine the existence of m6A modification on a specific target site as low as 100 amol under isothermal conditions. Detections of m6A can be performed on rRNA, mRNA, lincRNA, lncRNA and pre-miRNA from biological samples with naked-eye observations after dye incubation. Together, we provide a powerful tool for locus-specific detection of m6A, which can simply, quickly, sensitively, specifically, and visually determine putative m6A modification on RNA.