Search

Background. Reframing could be considered to represent individual and collective twofold values in social learning. When reframing is promoted, it benefits to generate learning processes involving collective inquiry occurring in inter and outer formal education. Digital games, popular as an application for the young generation, are considered a specific community in which social learning occurs by playing games. Serious games (SG) are a type of digital game involving educational purposes. Hence SG is selected as a suitable educational strategy to promote reframing. The previous research investigated features of a gameplay experience associated with reframing. This study develops further based on these game features to examine if/how they improved reframing capabilities. From an educational perspective, specific guidelines were proposed as the research findings to assist serious games in designing and applying game features to enhance the gameplay experience of reframing. Methods. An expert evaluation was used to assess the guidelines. The seven experts with experience in game design and teaching were chosen to review the designed guidelines. These experts were required to rate the guidelines using a 5-point Likert scale. All experts offered positive feedback on the finding. Results. Research data were analysed and validated via the Content Validity Index. The results showed that a total of 5 guidelines was remained and updated. Conclusion. These guidelines can be applied to assist students, game designers, related educators, and researchers who use games to promote reframing or require evidence to link game features with social learning.

The Community of Inquiry (COI) has become increasingly popular as a practical framework that promotes critical thinking and improves learning skills in online environments. In order to encourage the COI, this study proposes to investigate learning VALUEs based on teaching presence (TP), social presence (SP), and cognitive presence (CP), applying a VALUE creation framework as an assessment tool. A quantitative research approach was used to analyse 16 research works. Firstly, this investigation reviewed the activities of COIs as the vital link between COI and VALUE creation. Secondly, these activities were evaluated using Wenger et al.'s framework. The related results show that COI activities generate immediate, potential, and applied VALUEs, however, it does not positively promote reframing. The activities of TP positively promote immediate VALUE, especially its categories of promoting discourse and direct instruction. Most activities of SP promote both immediate and potential VALUEs, whose indicator of open communication positively creates immediate VALUE whereas the activities of CP promote multiple VALUEs. These findings address the research gap regarding how COI contributes VALUE by improving learning experiences in virtual environments and what determines the generation of the VALUEs. The benefits of this study will guide practitioners (teachers, online course developers, and instructional designers) who aim to design and match related activities of COI to maximise the VALUE creation.

Natural killer (NK) cells have shown good application prospects in adoptive cellular immunotherapy against cancer. However, due to its insufficient infiltration and low activity, the therapeutic effect of infused NK cells has been limited in solid tumors, such as colorectal cancer. It has been proved that tumor-produced chemokines regulate the migration of NK cells expressing corresponding chemokine receptors, and cytokines could enhance the antitumor activity of NK cells. In this study, we innovatively upregulated the expression of chemokine receptor CXC chemokine receptor 2 (CXCR2) and cytokine interleukin (IL)-2 on NK-92 cells using CRISPR-Cas9 gene-editing technology. We demonstrated that overexpressing CXCR2 and IL-2 promotes NK-92 cells to increasingly transfer into tumor sites and achieve stronger cell-killing and proliferation activity. Moreover, the inhibitory effects of gene-edited NK-92 cells on the growth of human colon cancer

Taking advantages of the catalytic activity of METTL3/METTL14 protein towards adenine methylation in RNA sequence and the specific digestion activity of MazF protein towards unmethylated RNA sequence containing ACA bases, a novel photoelectrochemical biosensor was constructed for simultaneous detection of RNA methylation, METTL3/METTL14 protein and MazF protein. MoSe

Transcription factor Yin Yang 1 (YY1) is upregulated in multiple tumors and plays essential roles in tumor proliferation and metastasis. However, the function of YY1 in breast cancer stemness remains unclear. Herein, we found that YY1 expression was negatively correlated with the overall survival and relapse-free survival of breast cancer patients and positively correlated with the expression of stemness markers in breast cancer. Overexpression of YY1 increased the expression of stemness markers, elevated CD44+CD24− cell sub-population, and enhanced the capacity of cell spheroid formation and tumor-initiation. In contrast, YY1 knockdown exhibited the opposite effects. Mechanistically, YY1 decreased microRNA-873-5p (miR-873-5p) level by recruiting histone deacetylase 4 (HDAC4) and HDAC9 to miR-873-5p promoter and thus increasing the deacetylation level of miR-873-5p promoter. Sequentially, YY1 activated the downstream PI3K/AKT and ERK1/2 pathways, which have been confirmed to be suppressed by miR-873-5p in our recent work. Moreover, the suppressed effect of YY1/miR-873-5p axis on the stemness of breast cancer cells was partially dependent on PI3K/AKT and ERK1/2 pathways. Finally, it was found that the YY1/miR-873-5p axis is involved in the chemoresistance of breast cancer cells. Our study defines a novel YY1/miR-873-5p axis responsible for the stemness of breast cancer cells., Graphical Abstract, Tumor recurrence and chemotherapy resistance often result from cancer stem cells. Guo et al. define a novel YY1/miR-873-5p axis responsible for the stemness of breast cancer cells. Moreover, the researchers found that the YY1/miR-873-5p axis is involved in the chemoresistance of breast cancer cells. The authors’ research indicates that YY1/miR-873-5p regulatory axis may be a potential molecular target for breast cancer therapy.

Given the improved photoactivity of Bi

Background The present study aimed to explore residues' properties interacting with HLA-A*02-restricted peptides on T-cell receptors (TCRs) and their effects on bond types of interaction and binding free energy. Methods We searched the crystal structures of HLA-A*02-restricted peptide-TCR complexes from the Protein Data Bank (PDB) database and subsequently collected relevant parameters. We then employed Schrodinger to analyze the bond types of interaction and Gromacs 2019 to evaluate the TCR-antigen peptide complex's molecular dynamics simulation. Finally, we compared the changes of bond types of interaction and binding free energy before and after residue substitution to ensure consistency of the conditions before and after residue substitution. Results The main sites on the antigen peptides that formed the intermolecular interaction [hydrogen bond (HB) and pi stack] with TCRs were P4, P8, P2, and P6. The hydrophobicity of the amino acids inside or outside the disulfide bond of TCRs may be related to the intermolecular interaction and binding free energy between TCRs and peptides. Residues located outside the disulfide bond of TCR α or β chains and forming pi stack force played favorable roles in the complex intermolecular interaction and binding free energy. The residues of the TCR α or β chains that interacted with peptides were replaced by alanine (Ala) or glycine (Gly), and their intermolecular binding free energy of the complex had been improved. However, it had nothing to do with the formation of HB. Conclusions The findings of this study suggest that the hydrophobic nature of the amino acids inside or outside the disulfide bonds on the TCR may be associated with the intermolecular interaction and binding between the TCR and polypeptide. The residues located outside the TCR α or β single-chain disulfide bond and forming the pi-stack force showed a beneficial effect on the intermolecular interaction and binding of the complex. In addition, the part of the residues on the TCR α or β single chain that produced bond types of interaction with the polypeptide after being replaced by Ala or Gly, the intermolecular binding free energy of the complex was increased, regardless of whether HB was formed.

NK cells are an important part of human immunity and are often used in clinical antitumor adoptive cell immunotherapy.However, the small number and low activity of NK cells in the tumor microenvironment limit its clinical efficacy.Studies have found that cytokines IL2 and IL12 play key roles in the proliferation of NK cells and the improvement of the activity of killing tumor cells, the toxic and side effects, however, are relatively large when the systemic application reaches the effective concentration. To study the effect of autocrine secretion of IL2 and IL12 on proliferation and anti-tumor activity of NK92 cell lines by editing NK cells' genes, and to construct NK92 cells with sustainable proliferation. Lentivirus was transfected into NK92 cells through the synergistic effect of crispr/cas9 localization gene editing technology and VP64 transcriptional activation domain to construct NK92-IL2-IL12 cell lines. The expression levels of IL2 and IL12 in the cell lines were detected by qPCR and ELISA.NK92 cell lines were inoculated into mouse tumor and the proliferation and anti-tumor effect of NK cells in tumor were observed. The vector plasmids IL2 and IL12 were successfully transferred into NK92 cells, which activated the up-regulated expression of IL2/IL12 in NK92, significantly increased the activity and lethality of NK92 cells, and sustained the proliferation of NK92 cells in mouse tumor.IL2 and IL12 can significantly promote the proliferation and killing activity of NK92 cell lines through autocrine, which have stronger anti-tumor activity than the wild-type ones, with continuous proliferation in the tumor.

The new military revolution in the new era is spurring the development of combat means and combat forces to unmanned, intelligent, and clustered direction and gradually forming new methods of warfare and winning mechanisms. Unmanned combat platforms represented by unmanned aerial vehicle (UAV), unmanned combat vehicle, unmanned surface vessel, and unmanned underwater vehicle have gradually become new research areas in which various military powers are committing to. Among the various unmanned combat platforms, UAVs achieve the fastest development and have the most operational applications. This paper gives a brief overview of UAVs and their applications and focuses on the advantages and problems of UAV and UAV cluster.

Successive infusion of natural killer cells is increasingly being explored as a treatment for cancer patients. The inadequate homing of natural killer cells into the tumor site resulted in the poor efficacy of natural killer cells on solid tumors. For the adoptive transfer of tumor-directed natural killer cell has been proved effective, it is hypothesized that there must be more association between the tumor-produced chemokines and the natural killer cells-expressed chemokine receptors. Increased CXCL12 and CCL21 could ameliorated colorectal cancer via generating an anti-tumor environment by preferentially attracting natural killer cells which expressed the chemokine receptor CXCR4 and CCR7. This study demonstrated that overexpressed CXCR4 and CCR7 on the surface of NK92 cell enhanced their migration to human colon cells. Moreover, the administration of such natural killer cells resulted in tumor shrinkage and a significantly increased survival of experimental mice when compared to ones undergoing the treatment of xenografts with natural killer cells expressing only the mock control. These suggested that chemokine receptor engineered natural killer cells could be a promising tool to improve adoptive tumor immunotherapy.

Background Breast cancer stem cells have self-renewal capability and are resistant to conventional chemotherapy. PD-L1 could promote the expression of stemness markers (OCT4 and Nanog) in breast cancer stem cells. However, the mechanisms by which PD-L1 regulates the stemness of breast cancer cells and PD-L1 is regulated in breast cancer cells are still unclear. Methods Lentivirus infection was used to construct stable cell lines. The correlation between PD-L1 and stemness markers expression was evaluated in clinical samples. Additionally, luciferase reporter assay combined with RNA-Fluorescence in situ hybridization (RNA-FISH) and RNA-binding protein immunoprecipitation (RIP) assays were used to verify the direct binding of miR-873 on PD-L1. Furthermore, flow cytometry, mammosphere formation combined with nude mouse tumor xenograft model were carried out to examine the effects of miR-873/PD-L1 axis on the stemness of breast cancer cells. Finally, MTT assay was performed to determine the effects of miR-873/PD-L1 axis on drug resistance. Findings PD-L1 expression was positively correlated with the expression of stemness markers, and overexpression of PD-L1 contributed to chemoresistance and stemness-like properties in breast cancer cells via activating PI3K/Akt and ERK1/2 pathways. Mechanistically, miR-873 inhibited PD-L1 expression through directly binding to its 3′-untranslated region (UTR), and miR-873 attenuated the stemness and chemoresistance of breast cancer cells which was dependent on PD-L1 and the downstream PI3K/Akt and ERK1/2 signaling. Notably, the promotion of PD-L1 on the stemness and chemoresistance was enhanced by recombinant PD-1 (rPD-1), this effect was attenuated by PD-1/PD-L1 inhibitor. Interpretation miR-873/PD-L1 regulatory axis might serve as a therapeutic target to enhance the chemo-sensitivity and eliminate the stemness of breast cancer cells. Fund This work was supported by the National Nature Science Foundation of China, No. 81702957, China Postdoctoral Science Foundation, No. 2017M620230, the Postdoctoral Research Funding Scheme of Jiangsu Province (2017), No. 1701197B, and the Priority Academic Program Development (PAPD) of Jiangsu Higher Education Institutions., Highlights • A critical role of PD-L1 in stemness and chemoresistance is proposed. • A negative miR-873/PD-L1 interaction was identified in breast cancer cells. • The mechanisms of miR-873/PD-L1 in stemness and chemoresistance were studied. • The results provide new insights for breast cancer progression and treatment.

Table S4. Sequences of siRNA against specific target in this study (DOC 32 kb)

Figure S1. CeRNET_CC promotes the stemness of MCF-7 cells in vitro. (A) The expression of CYP4Z2P and CYP4Z1 in MCF-7 and MCF-7-tumorsphere cells was detected by qRT-PCR. (B and C) The infection efficiency of MCF-7 cells with CYP4Z1- or CYP4Z2P-3′UTR stable overexpression (B) or knockdown (C) was detected by qRT-PCR. (D) Phase contrast images of mammospheres formed by stable expression cells depicted in B and C and quantification of spheres. (E) Representative FACS profile of cells described in B with CD24− and CD44+ markers. (F and G) The mRNA and protein expression of stemness markers (ALDH1, SOX2, OCT4 and Nanog) in cells described in B and C were examined by qRT-PCR and western blot analysis, respectively. The data are presented as the means ± SDs, n = 3, *P

Table S6. Sequences of primers used for Luciferase reporter assay. (DOC 46 kb)

Table S2. Primary antibodies used in this study. (DOC 35 kb)

Figure S4. The correlation between six2 expression and the survival of breast cancer patients. (A) KM-plotter survival curves showed the disease-free survival probability of patients separated into low and high six2 levels. (B) KM-plotter survival curves showed the OS survival probability of patients separated into low and high six2 level. (C and F) KM-plotter survival curves showed the RFS probability of patients separated into low and high six2 levels ( http://hgserver1.amc.nl/cgi-bin/r2/main.cgi ). (PDF 576 kb)

Table S5. Sequences of primers used for plasmid constructions. (DOC 40 kb)

Table S3. Sequences of primers used for ChIP qRT-PCR in this study. (DOCX 16 kb)

Figure S3. Transcriptional factor six2 promotes the expression of CYP4Z1 and the pseudogene CYP4Z2P. (A) Computational analysis of the CYP4Z2P and CYP4Z1 promoters showed potential binding sites for six2. (B and C) Fragments of the CYP4Z2P and CYP4Z1 promoters were cloned into the luciferase reporter vector pGL3. MCF-7 cells were co-transfected with six2 and the luciferase constructs or the control construct. 72 h later, luciferase activity was measured. (D) Relative luciferase activity was detected in MCF-7 cells co-transfected with the six2 overexpression vector and CYP4Z1 and CYP4Z2P promoter vectors with six2 binding sites or mutated six2 binding sites. (E and F) The expression of CYP4Z2P, CYP4Z1, and six2 in MCF-7-six2 (E) and 231-six2 (F) cells was examined by qRT-PCR. (G and H) CYP4Z1 protein expression in MCF-7-six2 (G) and MCF7-Plko-six2 (H) cells was detected by western blot. The data were presented as the means ± SDs, n = 3, **P

Background The expression of CYP4Z1 and the pseudogene CYP4Z2P has been shown to be specifically increased in breast cancer by our group and others. Additionally, we previously revealed the roles of the competitive endogenous RNA (ceRNA) network mediated by these genes (ceRNET_CC) in breast cancer angiogenesis, apoptosis, and tamoxifen resistance. However, the roles of ceRNET_CC in regulating the stemness of breast cancer cells and the mechanisms through which ceRNET_CC is regulated remain unclear. Methods Transcriptional factor six2, CYP4Z1-3′UTR, and CYP4Z2P-3′UTR were stably overexpressed or knocked down in breast cancer cells via lentivirus infection. ChIP-sequencing and RNA-sequencing analysis were performed to reveal the mechanism through which ceRNET_CC is regulated and the transcriptome change mediated by ceRNET_CC. Clinical samples were used to validate the correlation between six2 and ceRNET_CC. Finally, the effects of the six2/ceRNET_CC axis on the stemness of breast cancer cells and chemotherapy sensitivity were evaluated by in vitro and in vivo experiments. Results We revealed that ceRNET_CC promoted the stemness of breast cancer cells. Mechanistically, six2 activated ceRNET_CC by directly binding to their promoters, thus activating the downstream PI3K/Akt and ERK1/2 pathways. Finally, we demonstrated that the six2/ceRNET_CC axis was involved in chemoresistance. Conclusions Our results uncover the mechanism through which ceRNET_CC is regulated, identify novel roles for the six2/ceRNET_CC axis in regulating the stemness of breast cancer cells, and propose the possibility of targeting the six2/ceRNET_CC axis to inhibit breast cancer stem cell (CSC) traits.

Background Breast cancer stem cells have self-renewal capability and are resistant to conventional chemotherapy. PD-L1 could promote the expression of stemness markers (OCT4 and Nanog) in breast cancer stem cells. However, the mechanisms by which PD-L1 regulates the stemness of breast cancer cells and PD-L1 is regulated in breast cancer cells are still unclear. Methods Lentivirus infection was used to construct stable cell lines. The correlation between PD-L1 and stemness markers expression was evaluated in clinical samples. Additionally, luciferase reporter assay combined with RNA-Fluorescence in situ hybridization (RNA-FISH) and RNA-binding protein immunoprecipitation (RIP) assays were used to verify the direct binding of miR-873 on PD-L1. Furthermore, flow cytometry, mammosphere formation combined with nude mouse tumor xenograft model were carried out to examine the effects of miR-873/PD-L1 axis on the stemness of breast cancer cells. Finally, MTT assay was performed to determine the effects of miR-873/PD-L1 axis on drug resistance. Findings PD-L1 expression was positively correlated with the expression of stemness markers, and overexpression of PD-L1 contributed to chemoresistance and stemness-like properties in breast cancer cells via activating PI3K/Akt and ERK1/2 pathways. Mechanistically, miR-873 inhibited PD-L1 expression through directly binding to its 3′-untranslated region (UTR), and miR-873 attenuated the stemness and chemoresistance of breast cancer cells which was dependent on PD-L1 and the downstream PI3K/Akt and ERK1/2 signaling. Notably, the promotion of PD-L1 on the stemness and chemoresistance was enhanced by recombinant PD-1 (rPD-1), this effect was attenuated by PD-1/PD-L1 inhibitor. Interpretation miR-873/PD-L1 regulatory axis might serve as a therapeutic target to enhance the chemo-sensitivity and eliminate the stemness of breast cancer cells. Fund This work was supported by the National Nature Science Foundation of China, No. 81702957, China Postdoctoral Science Foundation, No. 2017M620230, the Postdoctoral Research Funding Scheme of Jiangsu Province (2017), No. 1701197B, and the Priority Academic Program Development (PAPD) of Jiangsu Higher Education Institutions.

RNA binding proteins (RBPs) are pivotal post-transcriptional regulators. RNPC1, an RBP, acts as a tumor suppressor through binding and regulating the expression of target genes in cancer cells. This study disclosed that RNPC1 expression was positively correlated with breast cancer patients' relapse-free and overall survival and that RNPC1 suppressed breast cancer cell metastasis. Mechanistically, RNPC1 promotes competing endogenous RNA (ceRNA) network crosstalk among STARD13, CDH5, HOXD10, and HOXD1 (STARD13-correlated ceRNA network), which we previously confirmed in breast cancer cells through stabilizing the transcripts and thus facilitating the expression of these four genes in breast cancer cells. Furthermore, RNPC1 overexpression restrained the promotion of STARD13, CDH5, HOXD10, and HOXD1 knockdown on cell metastasis. Notably, RNPC1 expression was positively correlated with CDH5, HOXD1, and HOXD10 expression in breast cancer tissues and attenuated adriamycin resistance. Taken together, these results identified that RNPC1 could inhibit breast cancer cell metastasis via promoting a STARD13-correlated ceRNA network.

Table S2. Sequences of primers used for qRT-PCR in this study (DOC 44Â kb)

Table S1. Sequences of siRNA against specific target in this study. (DOC 34Â kb)

Table S3. Primary antibodies used in this study. (DOC 36Â kb)

Table S4. Sequences of primers used for plasmid constructions. (DOC 41Â kb)

An effective optimum charging technique for lithium ion batteries using a universal voltage protocol (UVP) that can accommodate cell aging is presented here. This charging method demands less learning to varying state-of-health (SOH) conditions with potential to improve charging efficiency and cycle life. The simplicity of UVP makes the implementation easier than the conventional constant current–constant voltage (CC–CV)-based methods. Here, the mathematical formulation, optimization targets (e.g. minimal time) and constraints (terminal voltages and other instrumental and cell electrochemistry-limited ones) are explained from the protocol design considerations. An equivalent circuit model was used and its parameters derived from the analysis of test data, which could yield a nonlinear varying current profile (VCP) by simulation and a genetic algorithm-based optimization. Both UVP and VCP were used in the validation to illustrate better charging efficiency and capacity retention, which showed a much improved cycle life.