Expression and distribution of PPP2R5C gene in leukemia

<p>Abstract</p> <p>Background</p> <p>Recently, we clarified at the molecular level novel chromosomal translocation t(14;14)(q11;q32) in a case of Sézary syndrome, which caused a rearrangement from TRAJ7 to the <it>PPP2R5C </it>gene. <it>PPP2R5C </it>is one of the regulatory B subunits of protein phosphatase 2A (PP2A). It plays a crucial role in cell proliferation, differentiation, and transformation. To characterize the expression and distribution of five different transcript variants of the <it>PPP2R5C </it>gene in leukemia, we analyzed the expression level of <it>PPP2R5C </it>in peripheral blood mononuclear cells from 77 patients with <it>de novo </it>leukemia, 26 patients with leukemia in complete remission (CR), and 20 healthy individuals by real-time PCR and identified the different variants of <it>PPP2R5C </it>by RT-PCR.</p> <p>Findings</p> <p>Significantly higher expression of <it>PPP2R5C </it>was found in AML, CML, T-ALL, and B-CLL groups in comparison with healthy controls. High expression of <it>PPP2R5C </it>was detected in the B-ALL group; however, no significant difference was found compared with the healthy group. The expression level of <it>PPP2R5C </it>in the CML-CR group decreased significantly compared with that in the <it>de novo </it>CML group and was not significantly different from the level in the healthy group. By using different primer pairs that covered different exons, five transcript variants of <it>PPP2R5C </it>could be identified. All variants could be detected in healthy samples as well as in all the leukemia samples, and similar frequencies and distributions of <it>PPP2R5C </it>were indicated.</p> <p>Conclusions</p> <p>Overexpression of <it>PPP2R5C </it>in T-cell malignancy as well as in myeloid leukemia cells might relate to its proliferation and differentiation. Investigation of the effect of target inhibition of this gene might be beneficial to further characterization of molecular mechanisms and targeted therapy in leukemia.</p

Comprehensive succinylome analyses reveal that hyperthermia upregulates lysine succinylation of annexin A2 by downregulating sirtuin7 in human keratinocytes

Background and Objectives: Local hyperthermia at 44°C can clear multiple human papillomavirus (HPV)-infected skin lesions (warts) by targeting a single lesion, which is considered as a success of inducing antiviral immunity in the human body. However, approximately 30% of the patients had a lower response to this intervention. To identify novel molecular targets for anti-HPV immunity induction to improve local hyperthermia efficacy, we conducted a lysine succinylome assay in HaCaT cells (subjected to 44°C and 37°C water baths for 30 min). Methods: The succinylome analysis was conducted on HaCaT subjected to 44°C and 37°C water bath for 30 min using antibody affinity enrichment together with liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results were validated by western blot (WB), immunoprecipitation (IP), and co-immunoprecipitation (Co-IP). Then, bioinformatic analysis including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, motif characterization, secondary structure, and protein–protein interaction (PPI) was performed. Results: A total of 119 proteins with 197 succinylated sites were upregulated in 44°C-treated HaCaT cells. GO annotation demonstrated that differential proteins were involved in the immune system process and viral transcription. Succinylation was significantly upregulated in annexin A2. We found that hyperthermia upregulated the succinylated level of global proteins in HaCaT cells by downregulating the desuccinylase sirtuin7 (SIRT7), which can interact with annexin A2. Conclusions: Taken together, these data indicated that succinylation of annexin A2 may serve as a new drug target, which could be intervened in combination with local hyperthermia for better treatment of cutaneous warts

Altered expression of the TCR signaling related genes CD3 and FcεRIγ in patients with aplastic anemia

<p>Abstract</p> <p>Background</p> <p>Aplastic anemia (AA) is characterized by pancytopenia and bone marrow hypoplasia, which results from immune-mediated hematopoiesis suppression. Understanding the pathophysiology of the immune system, particularly T cells immunity, has led to improved AA treatment over the past decades. However, primary and secondary failure after immunosuppressive therapy is frequent. Thus, knowledge of the immune mechanisms leading to AA is crucial to fundamentally understand the disease.</p> <p>Findings</p> <p>To elucidate the T cell receptor (TCR) signal transduction features in AA, the expression levels of CD3γ, δ, ε and ζ chain and FcεRIγ genes, which are involved in TCR signal transduction, and the negative correlation of the expression levels between the CD3ζ and FcεRIγ genes in T cells from peripheral blood mononuclear cells (PBMCs) were analyzed. Real-time RT-PCR using the SYBR Green method was used to detect the expression level of these genes in PBMCs from 18 patients with AA and 14 healthy individuals. The β2microglobulin gene (β2M) was used as an endogenous reference. The expression levels of the CD3γ, CD3δ, CD3ε and CD3ζ genes in patients with AA were significantly increased compared to a healthy control group, whereas the FcεRIγ gene expression level was significantly decreased in patients with AA in comparison with the healthy control group. Moreover, the negative correlation of the expression levels between the CD3ζ and FcεRIγ genes was lost.</p> <p>Conclusions</p> <p>To our knowledge, this is the first report of the CD3γ, CD3δ, CD3ε, CD3ζ and FcεRIγ gene expression in patients with AA. The abnormally expressed TCR signaling related genes may relate to T cells dysfunction in AA.</p

Optimisation for image salient object detection based on semantic‐aware clustering and CRF

State‐of‐the‐art optimisation methods for salient object detection neglect that saliency maps of different images usually show different imperfections. Therefore, the saliency maps of some images cannot achieve effective optimisation. Based on the observation that the saliency maps of semantically similar images usually show similar imperfections, the authors propose an optimisation method for salient object detection based on semantic‐aware clustering and conditional random field (CRF), named CCRF. They first cluster the training images into some clusters using the image semantic features extracted by using a deep convolutional neural network model for image classification. Then for each cluster, they use a CRF to optimise the saliency maps generated by existing salient object detection methods. A grid search method is used to compute the optimal weights of the kernels of the CRF. The saliency maps of the testing images are optimised by the corresponding CRFs with the optimal weights. The experimental results with 13 typical salient object detection methods on four datasets show that the proposed CCRF algorithm can effectively improve the results of a variety of image salient object detection methods and outperforms the compared optimisation methods

Study on the strata displacement law of overlapping shield tunnels with different overlapping angles

Based on the overlapping tunnel of Tel Aviv Red Line Light Rail Project in Israel, this study established a three-dimensional numerical model by using Rhino6 and FLAC3D software on the basis of analysis of on-site monitoring data, to investigate the ground settlement and stratum displacement laws of shield overlapping tunnel at different overlapping angles.Results show that when the two tunnels gradually transit from horizontal to overlapping state, with the increase of overlapping angle of the two tunnels, the accumulated settlement at the measuring points in the middle of the two tunnels increases, but the accumulated settlement is not only affected by overlapping angle, but also closely related to the stratum traversing situation.Under the stratum condition of the project, when the overlapping angle is 60°, the accumulated surface settlement reaches the maximum because both tunnels pass through the loose reservoir K1 stratum.When the overlapping angle of the rear-built tunnel is less than 45° after excavation, the width of the settlement trough increases.When the overlapping angle is larger than 45° and smaller than 60°, the width of the settlement trough remains unchanged.The width of settlement trough decreases when the overlapping angle is larger than 60°.When the overlapping angle reaches 90°, the settlement trough is symmetrical on both sides.After excavation of tunnels on both sides, when the overlapping angle of the tunnels reaches 75°, the strata on both sides change the direction of movement.This research offer reference for the design and construction of overlapping tunnels

Viral delivery of multiple miRNA promotes retinal ganglion cell survival and functional preservation after optic nerve crush injury

Bone marrow mesenchymal stem cell (BMSC)-derived small extracellular vesicles (sEV) but not fibroblast sEV provide retinal ganglion cell (RGC) neuroprotection both in vitro and in vivo, with miRNAs playing an essential role. More than 40 miRNAs were more abundant in BMSC-sEV than in fibroblast-sEV. The purpose of this study was to test the in vitro and in vivo neuroprotective and axogenic properties of six candidate miRNAs (miR-26a, miR-17, miR-30c-2, miR-92a, miR-292, and miR-182) that were more abundant in BMSC-sEV than in fibroblast-sEV. Adeno-associated virus (AAV)-2 expressing a combination of three of the above candidate miRNAs were added to heterogenous adult rat retinal cultures or intravitreally injected into rat eyes one week before optic nerve crush (ONC) injury. Survival and neuritogenesis of βIII-tubulin+ RGCs was assessed in vitro, as well as the survival of RBPMS+ RGCs and regeneration of their axons in vivo. Retinal nerve fiber layer thickness (RNFL) was measured to assess axonal density whereas positive scotopic threshold response electroretinography amplitudes provided a readout of RGC function. Qualitative retinal expression of PTEN, a target of several of the above miRNAs, was used to confirm successful miRNA activity. AAV2 reliably transduced RGCs in vitro and in vivo. Viral delivery of miRNAs in vitro showed a trend towards neuroprotection but remained insignificant. Delivery of selected combinations of miRNAs (miR-17-5p, miR-30c-2 and miR-92a; miR-92a, miR-292 and miR-182) before ONC provided significant therapeutic benefits according to the above measurable endpoints. However, no single miRNA appeared to be responsible for the effects observed, whilst positive effects observed appeared to coincide with successful qualitative reduction in PTEN immunofluorescence in the retina. Viral delivery of miRNAs provides a possible neuroprotective strategy for injured RGCs that is conducive to therapeutic manipulation

Complete and Resilient Documentation for Operational Medical Environments Leveraging Mobile Hands-free Technology in a Systems Approach: Experimental Study

BACKGROUND: Prehospitalization documentation is a challenging task and prone to loss of information, as paramedics operate under disruptive environments requiring their constant attention to the patients. OBJECTIVE: The aim of this study is to develop a mobile platform for hands-free prehospitalization documentation to assist first responders in operational medical environments by aggregating all existing solutions for noise resiliency and domain adaptation. METHODS: The platform was built to extract meaningful medical information from the real-time audio streaming at the point of injury and transmit complete documentation to a field hospital prior to patient arrival. To this end, the state-of-the-art automatic speech recognition (ASR) solutions with the following modular improvements were thoroughly explored: noise-resilient ASR, multi-style training, customized lexicon, and speech enhancement. The development of the platform was strictly guided by qualitative research and simulation-based evaluation to address the relevant challenges through progressive improvements at every process step of the end-to-end solution. The primary performance metrics included medical word error rate (WER) in machine-transcribed text output and an F1 score calculated by comparing the autogenerated documentation to manual documentation by physicians. RESULTS: The total number of 15,139 individual words necessary for completing the documentation were identified from all conversations that occurred during the physician-supervised simulation drills. The baseline model presented a suboptimal performance with a WER of 69.85% and an F1 score of 0.611. The noise-resilient ASR, multi-style training, and customized lexicon improved the overall performance; the finalized platform achieved a medical WER of 33.3% and an F1 score of 0.81 when compared to manual documentation. The speech enhancement degraded performance with medical WER increased from 33.3% to 46.33% and the corresponding F1 score decreased from 0.81 to 0.78. All changes in performance were statistically significant (P\u3c.001). CONCLUSIONS: This study presented a fully functional mobile platform for hands-free prehospitalization documentation in operational medical environments and lessons learned from its implementation