Rescue of IFN-γ production following CD244/2B4 receptor blocking.

<p>(A) Representative flow cytometric plots showing IFN-γ production in stimulated CD3⁺CD4⁺ T cells when incubated with isotype control antibody (ISO) or blocking anti-CD244/2B4 antibody clone eBioPP35 (BL). (B to D) Blocking of CD244/2B4 signaling resulted in significantly higher frequencies of IFN-γ-producing CD4⁺ T cells (B), increased level of IFN-γ production in CD3⁺CD4⁺ T cells as determined by flow cytometry (C), and elevated numbers of spot forming units in ELISPOT assay (D), as compared with those incubated with isotype control antibody. Paired t-test was used for statistical analysis. **: p<0.01.</p

Activation of CD244/2B4 signaling resulted in significantly decreased IFN-γ production.

<p>(A) Representative flow cytometric plots showing IFN-γ production in CD3⁺CD4⁺ T cells incubated with isotype control antibody (ISO) or with anti-CD244/2B4 antibody clone C1.7 (CK). (B to D) Activation of CD244/2B4 signaling by cross-linking led to significantly lower frequencies of IFN-γ-producing CD4⁺ T cells (B), decreased expression of IFN-γ as determined by flow cytometry (C), and reduced numbers of spot forming units in ELISPOT assay (D). Paired t-test was used for statistical analysis. *: p<0.05; **: p<0.01.</p

Representative flow cytometric plots showing gating strategy and expression of CD244/2B4.

<p>(A) Viable lymphocytes in PBMCs were gated based on characteristic forward and side scatter profiles. (B) The gate depicting CD4/CD244 expression was defined by both control isotype staining and CD244 expression on CD3⁻CD4⁻ cells that showed clearly two cell subsets with positive and negative staining of CD244. (C and D) CD4⁺ T cells were defined as CD3⁺CD4⁺ cells (C) and CD244/2B4 expression on total CD4⁺ T cells was shown (D). (E and F) Antigen-responsive CD4<b>⁺</b> T cells were identified by appearance of activation marker CD69 in the CD3⁺CD4⁺ cell population (E) and CD244/2B4 expression on <i>M. tuberculosis</i> antigen-responsive CD4⁺ T cells was shown (F). Isotype: isotype antibody control; LI: latent TB infection; TB: active TB patients.</p

miR-582-5p expression detected by real-time RT-PCR.

<p>(A) Relative expression of miR-582-5p in CD3⁻CD33⁺, CD3⁻CD33⁻, CD3⁺ cell subsets and in PBMCs from patients with active TB (n = 3). miR-582-5p was mainly expressed in CD3⁻CD33⁺ cells. (B) Patients with active TB had significantly higher expression of miR-582-5p as compared with healthy controls. The relative expression of miR-582-5p, as defined by mean -ΔCt, was normalized against U6 snRNA. Two-tailed unpaired t-test was used for statistical analysis between groups.</p

List of predicted target genes of miR-582-5p.

<p>List of predicted target genes of miR-582-5p.</p

Frequencies and apoptosis of CD14⁺ peripheral blood monocytes in patients with active TB and healthy controls.

<p>(A) Representative flow cytometric plots showing gating strategy and percentage of monocytes (left panel). Monocytes were defined by high CD14 expression (gate BV). Patients with active TB had significantly elevated frequency of CD14⁺ monocytes compared with healthy controls. Two-tailed unpaired t-test was used for statistical analysis between groups. (B) Representative flow cytometric plots showing apoptosis of monocytes (upper panel). PBMCs from patients with active TB and healthy controls were incubated with RPMI-1640 contained 2% FBS for 24 h at 37°C, and cells were stained with fluorochrome-labeled anti-human CD14, Annexin V and PI. Cells that were positive for Annexin V were defined as apoptosis. Monocytes from patients with active TB had significantly lower percentage of apoptotic cells than that from healthy controls. Mann-Whitney test was used for statistical analysis between groups.</p

<i>FOXO1</i> is a direct target of miR-582-5p.

<p>(A) Alignment between the predicted miR-582-5p target site of <i>FOXO1</i> 3′UTR region and miR-582-5p. (B) Real time RT-PCR and western blot analysis showing <i>FOXO1</i> mRNA and protein expression levels in THP-1 cells transfected with miR-582-5p mimics or mimics NC, respectively. (C) Co-transfection of miR-582-5p mimics/mimics NC and <i>FOXO1</i> 3′UTR-luciferase reporter vector into HEK-293T cells demonstrated that significant decrease in luciferase activity was only found in reporter vector that contained a wild type sequence (FOXO1-3′UTR-wt), not in vector that contained a mutant sequence (FOXO1-3′UTR-mut) within the miR-582-5p binding site. Values were presented as relative luciferase activity after normalization to Renilla luciferase activity. FOXO1-3′UTR-wt: pMIR-FOXO1-3′UTR-wt vector; FOXO1-3′UTR-mut: pMIR-FOXO1-3′UTR-mut vector. (D) Representative flow cytometric plots showing apoptotic ratio of THP-1 transfected with <i>FOXO1</i> siRNA or negative controls of siRNA (siRNA NC) (left panel). The apoptotic percentage of THP-1 cells transfected with <i>FOXO1</i> siRNA were significantly lower than those transfected with negative control siRNA (siRNA NC) (<i>p</i><0.01).</p

Inhibition of apoptosis in monocytes by miR-582-5p.

<p>(A) Representative flow cytometric plots showing apoptotic percentage of THP-1 cells transfected with miR-582-5p mimics or negative control of mimics (mimics NC). (B) The apoptotic percentage of THP-1 cells transfected with miR-582-5p mimics was significantly lower than those transfected with negative control of microRNA mimics (mimics NC).</p

Demographic and clinical characteristics of patients with active TB and controls.

<p>Note: ¹ Erythrocyte sedimentation rate; ²The absolute number of monocytes was determined by white blood cell count; ³% of CD14⁺ Monocytes was determined by flow cytometry.</p

V₂O₅ Nanowire Composite Paper as a High-Performance Lithium-Ion Battery Cathode

Ultralong, as long as ∼1 mm, orthorhombic vanadium pentoxide (V₂O₅) nanowires were synthesized using a hydrothermal method. Free-standing and binder-free composite paper was prepared on a large scale by a two-step reduction method using free-standing V₂O₅ nanowires as the skeleton and reduced graphene oxide (rGO) nanosheets as the additive. Such a free-standing V₂O₅/rGO composite paper as a cathode for lithium ion batteries possesses both structural integrity and extraordinary electrochemical performance. The reversible specific areal capacity of the V₂O₅/rGO composite paper electrode is 885 μAh/cm² at 0.09 mA/cm², much higher than that of the pure V₂O₅ nanowire paper electrode (570 μAh/cm²). It also shows excellent cycling performance at high rates with 30.9% loss of its initial capacities after 1000 cycles at a current rate of 0.9 mA/cm². The excellent performance was attributed to the improved electronic conductivity and Li⁺ ion transport from the rGO addition