To improve the diagnosis and treatment of Mycoplasma pneumoniae (Mp) infection and reduce the misuse of antibiotics, we sought to establish a loop-mediated isothermal amplification (LAMP) assay for ...rapid detection of Mp.Six primers specific for the Mp P1 gene were designed, and the LAMP method was used to rapidly detect Mp. The sensitivity of the LAMP method was determined by serial dilution of the standard Mp strain FH (standard strains of Mycoplasma pneumoniae). Specificity was assessed with 17 common pathogenic microorganisms in the respiratory tract. Patient samples were collected from the Department of Respiratory and Critical Care Medicine at the 307th Hospital of Chinese People's Liberation Army from March 2016 to May 2017, examined prospectively, and compared with diagnosis by quantitative real-time polymerase chain reaction (qRT-PCR).The LAMP assay for Mp detection can be completed within 60 minutes. The minimum detection limit was 39 pg/μL, and no cross-reaction was observed with 17 common respiratory tract pathogens. Of the 125 clinical specimens tested, 43 cases were positive by LAMP assay, and 40 cases were positive by qRT-PCR (P = .162). All 43 samples determined as positive by LAMP test were confirmed to be Mp by Mp P1 protein sequencing.The LAMP assay is suitable for rapid detection of Mp. It has high sensitivity and specificity, and the detection results are not inferior to those of qRT-PCR.
This research studies the effects of axial preload on nonlinear dynamic characteristics of a flexible rotor supported by angular contact ball bearings. A dynamic model of ball bearings is improved ...for modeling a five-degree-of-freedom rotor bearing system. The predicted results are in good agreement with prior experimental data, thus validating the proposed model. With or without considering unbalanced forces, the Floquet theory is employed to investigate the bifurcation and stability of system periodic solution. With the aid of Poincarè maps and frequency response, the unstable motion of system is analyzed in detail. Results show that the effects of axial preload applied to ball bearings on system dynamic characteristics are significant. The unstable periodic solution of a balanced rotor bearing system can be avoided when the applied axial preload is sufficient. The bifurcation margins of an unbalanced rotor bearing system enhance markedly as the axial preload increases and relates to system resonance speed.
Acute lung injury and acute respiratory distress syndrome (ALI/ARDS) refer to acute and progressive hypoxic respiratory failure caused by non-cardiogenic factors, which is a common condition ...occurring in critically ill patients with widespread pulmonary inflammation. Use of a single medication or target cannot treat ALI/ARDS. Mesenchymal stem cells (MSCs) and FTY720, as an analogue of sphingosine-1-phosphate (S1P), can mitigate lipopolysaccharide (LPS)-induced inflammatory lung injury.
In this investigation, the clinical efficacy of MSCs alone, FTY720 alone, and a MSC and FTY720 combination in the treatment of LPS-induced lung injury was evaluated in mouse models. The experimental results demonstrated that both MSCs and FTY720 alleviate lung injuries in mice. The combined application of MSCs and FTY720 yielded higher clinical efficacy in mitigating lung injuries compared with use of MSCs or FTY720 alone. Transcriptomic analysis was performed using an Agilent gene expression chip. By analyzing the differences in gene expression of lung tissues between treated and non-treated ALI/ARDS mice, Gene Ontology and Pathway terms related to ALI/ARDS treatment were identified. Moreover, the target genes which might play a pivotal role in the treatment of ALI/ARDS were also detected, thus providing a theoretical basis for multi-target or multi-drug combined treatment of ALI/ARDS and lay a solid foundation for clinical treatment of ALI/ARDS.
•This investigation analyze the differences in gene expression of lung tissues between MSCs, FTY720, MSC and FTY720 combination treated ALI/ARDS mice.•Gene Ontology and Pathway terms related to ALI/ARDS treatment were identified.•The target genes which might play a pivotal role in the treatment of ALI/ARDS were also detected.•This investigation provide a theoretical basis for multi-target or multi-drug combined treatment of ALI/ARDS and lay a solid foundation for clinical treatment of ALI/ARDS.
Chronic skin wounds are often associated with multidrug-resistant bacteria, impeding the healing process. Bacteriophage (phage) therapy has been revitalized as a promising strategy to counter the ...growing concerns of antibiotic resistance. However, phage monotherapy also faces several application drawbacks, such as a narrow host spectrum, the advent of resistant phenotypes and poor stability of phage preparations. Phage-antibiotic synergistic (PAS) combination therapy has recently been suggested as a possible approach to overcome these shortcomings. In the present study, we employed a model PAS combination containing a vB_AbaM-IME-AB2 phage and colistin to develop stable wound dressings of PAS to mitigate infections associated with
. A set of thermosensitive hydrogels were synthesized with varying amounts of Pluronic® F-127 (PF-127 at 15, 17.5 and 20 w/w%) modified with/without 3 w/w% hydroxypropyl methylcellulose (HPMC). Most hydrogel formulations had a gelation temperature around skin temperature, suitable for topical application. The solidified gels were capable of releasing the encapsulated phage and colistin in a sustained manner to kill bacteria. The highest bactericidal effect was achieved with the formulation containing 17.5% PF-127 and 3% HPMC (F5), which effectively killed bacteria in both planktonic (by 5.66 log) and biofilm (by 3 log) states and inhibited bacterial regrowth. Good storage stability of F5 was also noted with negligible activity loss after 9 months of storage at 4 °C. The
antibacterial efficacy of the F5 hydrogel formulation was also investigated in a pork skin wound infection model, where it significantly reduced the bacterial burden by 4.65 log. These positive outcomes warrant its further development as a topical PAS-wound dressing.
Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are major causes of respiratory failure, but currently, no effective pharmacotherapy exists for these disorders. Alveolar ...macrophages play a critical role in both the acute/initial phase and chronic/resolving phase of ALI, rendering them a potential therapeutic target. Interleukin-4 (IL-4), a Th2 cytokine, not only directly inhibits the secretion of pro-inflammatory factors from macrophages but also drives macrophages to the anti-inflammatory and tissue remodeling M2 type. However, the short half-life of IL-4 in vivo hampers its effect on disease treatment. In this study, macrophages secreting IL-4 (M-IL-4) were established and used to treat ALI through pulmonary macrophage transplantation (PMT). The results showed that highly sustained levels of IL-4 and M2 macrophage markers were detected in mice lungs following pulmonary M-IL-4 transplantation. Furthermore, PMT improved the therapeutic effect by reducing lung inflammation, alleviating tissue injury, reducing alveolar macrophages necrotic cell death, and decreasing mortality in mice with ALI. These results suggest an efficient macrophage-based protein drug delivery strategy, and for the first time, prove the feasibility and efficacy of PMT in ALI treatment.
Acute lung injury (ALI), a persistent lung inflammatory response syndrome, may evolve into acute respiratory distress syndrome (ARDS). Characterized by rapid onset, critical features, and a complex ...etiology, ALI remains a challenging critical respiratory disease. Recently, mesenchymal stem cells (MSCs) have provided a new solution for the treatment of ALI. We built a lipopolysaccharide (LPS)-induced ALI model in mice. After treatment with human umbilical cord mesenchymal stem cells (hUC-MSCs), FTY720, or a combination of hUC-MSCs and FTY207, the lung inflammatory response was apparently attenuated. To understand the mechanism underlying MSCs treatment of ALI at the genetic level, significant differentially expressed long non-coding RNAs (lncRNAs) between the treatment and model groups were analyzed using microarray technology. Moreover, genetic gene prediction, gene ontology (GO) analysis, pathway analysis, and transcription factor (TF) prediction were carried out. The results showed that a total of 66 lncRNAs were differentially expressed in all three treatment groups, including 8 up-regulated and 58 down-regulated lncRNAs. LncRNA A_30_P01029806 and A_30_P01029194, which were down-regulated, were involved in the signaling pathways closely related to ALI. Through further TF analysis, we identified several significant TFs which lay a foundation for revealing the mechanism underlying lncRNAs treatment of ALI. LncRNA A_30_P01029806 and A_30_P01029194 may serve as candidate biomarkers in the diagnosis and treatment of ALI.
•66 lncRNAs were differentially expressed in all three treatment groups.•Two lncRNAs were involved in the signaling pathways closely related to ALI.•Several significant TFs related to the two lncRNAs were identified through TF prediction.•The two lncRNAs may serve as candidate biomarkers in the diagnosis and treatment of ALI.
Bacteriophage (phage) therapy, exploiting phage which are the natural enemies of bacteria, has been reintroduced to treat multidrug-resistant (MDR) bacterial infections. However, some intrinsic ...drawbacks of phage are overshadowing its clinical uses, particularly the narrow host spectrum and rapid emergence of resistance upon treatment. The use of phage and antibiotic combination exhibiting synergistic bacterial killing (termed phage-antibiotic synergy or PAS) has therefore been proposed. It is well-reported that the types and doses of phage and antibiotic are critical in achieving PAS. Comparatively, the impacts of treatment sequences are less explored. Here, we took an Acinetobacter baumannii phage vB_AbaM-IME-AB2 and colistin as a model PAS combination to elucidate the order effects in-vitro. While applying phage 8 h before colistin treatment demonstrated the greatest antibacterial synergy, it failed to prevent phage resistance development. On the other hand, simultaneous and antibiotic first followed by phage treatments could effectively suppress/delay resistance development. Between them, the simultaneous application demonstrated superior antibacterial and antibiofilm activities. Further in-vivo investigation is required to confirm the impacts of treatment sequences on the PAS application.
Background
Three-dimensional (3D) chromatin architecture frequently altered in cancer. However, its changes during the pathogenesis of hepatocellular carcinoma (HCC) remained elusive.
Methods
Hi-C ...and RNA-seq were applied to study the 3D chromatin landscapes and gene expression of HCC and ANHT. Hi-C Pro was used to generate genome-wide raw interaction matrices, which were normalized via iterative correction (ICE). Moreover, the chromosomes were divided into different compartments according to the first principal component (E1). Furthermore, topologically associated domains (TADs) were visualized via WashU Epigenome Browser. Furthermore, differential expression analysis of ANHT and HCC was performed using the DESeq2 R package. Additionally, dysregulated genes associated with 3D genome architecture altered were confirmed using TCGA, qRT-PCR, immunohistochemistry (IHC), etc.
Results
First, the intrachromosomal interactions of chr1, chr2, chr5, and chr11 were significantly different, and the interchromosomal interactions of chr4–chr10, chr13–chr21, chr15–chr22, and chr16–chr19 are remarkably different between ANHT and HCC, which resulted in the up-regulation of
TP53I3
and
ZNF738
and the down-regulation of
APOC3
and
APOA5
in HCC. Second, 49 compartment regions on 18 chromosomes have significantly switched (A–B or B–A) during HCC tumorigenesis, contributing to up-regulation of
RAP2A
. Finally, a tumor-specific TAD boundary located on chr5: 6271000–6478000 and enhancer hijacking were identified in HCC tissues, potentially associated with the elevated expression of
MED10
, whose expression were associated with poor prognosis of HCC patients.
Conclusion
This study demonstrates the crucial role of chromosomal structure variation in HCC oncogenesis and potential novel biomarkers of HCC, laying a foundation for cancer precision medicine development.
ALI/ARDS remain the main reason of morbidity and mortality in the critically ill. Studies have indicated that human umbilical cord mesenchymal stem cells (hUC-MSCs) can be useful in the treatment of ...ALI/ARDS. Sphingosine-1-phosphate (S1P) and its analog FTY720 significantly reduce lipopolysaccharide (LPS)-induced lung edema and inflammatory lung injury. This study aimed to assess the therapeutic effects of hUC-MSCs combined with FTY720 in an LPS-induced murine model of ALI. Eight-week-old female C57BL/6 mice were divided into a normal control group, an LPS group, an hUC-MSC group, an FTY720 group, and an hUC-MSCs+FTY720 group randomly. At 24 hours post injury, mice were administrated hUC-MSCs via the tail vein and/or intraperitoneally injected with FTY720. We assessed histopathology and histologic scores, lung wet/dry weight ratio, micro-CT scans, and total protein in the bronchoalveolar lavage fluid (BALF), as well as cytokines in the BALF at 48 h post injury. All treatment groups showed higher survival rates and attenuated lung injuries. The hUC-MSCs+FTY720 group yielded better results than hUC-MSCs or FTY720 alone. While the underlying mechanism requires further study, we anticipate that combination therapy of hUC-MSCs and FTY720 could be an effective strategy for ALI.
Influenza virus (IAV) infection is a major cause of severe respiratory illness that affects almost every country in the world. IAV infections result in respiratory illness and even acute lung injury ...and death, but the underlying mechanisms responsible for IAV pathogenesis have not yet been fully elucidated. In this study, the basic fibroblast growth factor 2 (FGF2) level was markedly increased in H1N1 virus-infected humans and mice. FGF2, which is predominately derived from epithelial cells, recruits and activates neutrophils via the FGFR2-PI3K-AKT-NFκB signaling pathway. FGF2 depletion or knockout exacerbated influenza-associated disease by impairing neutrophil recruitment and activation. More importantly, administration of the recombinant FGF2 protein significantly alleviated the severity of IAV-induced lung injury and promoted the survival of IAV-infected mice. Based on the results from experiments in which neutrophils were depleted and adoptively transferred, FGF2 protected mice against IAV infection by recruiting neutrophils. Thus, FGF2 plays a critical role in preventing IAV-induced lung injury, and FGF2 is a promising potential therapeutic target during IAV infection.
PDF
