Well-Wrapped Li-Rich Layered Cathodes by Reduced Graphene Oxide towards High-Performance Li-Ion Batteries

Layered lithium-rich manganese oxide (LLO) cathode materials have attracted much attention for the development of high-performance lithium-ion batteries. However, they have suffered seriously from disadvantages, such as large irreversible capacity loss during the first cycle, discharge capacity decaying, and poor rate performance. Here, a novel method was developed to coat the surface of 0.4Li2MnO3∙0.6LiNi1/3Co1/3Mn1/3O2 cathode material with reduced graphene-oxide (rGO) in order to address these drawbacks, where a surfactant was used to facilitate the well-wrapping of rGO. As a result, the modified LLO (LLO@rGO) cathode exhibits superior electrochemical performance including cycling stability and rate capability compared to the pristine LLO cathode. In particular, the LLO@rGO with a 0.5% rGO content can deliver a high discharge capacity of 166.3 mAh g−1 at a 5C rate. The novel strategy developed here can provide a vital approach to inhibit the undesired side reactions and structural deterioration of Li-rich cathode materials, and should be greatly useful for other cathode materials to improve their electrochemical performance

The sequences of chemically synthesized oligos.

a<p>The selected miRNAs were chemically synthesized in the form of small interfering RNA (siRNA) duplexes.</p>*<p>the bold and underlined letters were the mutation sites of miRNA.</p

MiR-140 increased the sensitivity of AECs to PTX.

<p>A: Morphological characteristics. Scale bars: 150 µm. TGF-β1 was added into miR-140- or ASO-140-transfected A549 cells for 24 h prior to PTX (50 nM) treatment. A549 cells treated with TGF-β1 or ASO-140 presented a fibroblast-like morphology, and the fibroblast-like morphology was reversed to epithelial-like characteristics in PTX- or PTX+miR-140-treated cultures. B: Western blot analysis. The levels of vimentin, Smad3 and p-Smad3 levels were increased and E-cadherin was downregulated in TGF-β1-treated A549 cells, which were reversed in PTX-, or PTX+miR-140-treated cells, especially in both PTX and miR-140-treated cultures. n = 3 replicates.</p

MiR-140 suppresses the TGF-β1/Smad3 pathway in A549 cells.

<p>A: Fluorescence microscope detection. Scale bars: 150 µm. B: Flow cytometry analysis. n = 3 replicates. Control, cells untreated with miRNA or plasmid; GFP-Smad3, cells treated with pcDNA-GFP-UTR; D-NC, cells treated with both D-NC (miR-140 mutation control) and pcDNA-GFP-UTR plasmid; S-NC, cells treated with both S-NC (the single-stranded RNA control) and pcDNA-GFP-UTR; miR-140, cells treated with both miRNA-140 and pcDNA-GFP-UTR; ASO-140, cells treated with both ASO-140 and pcDNA-GFP-UTR. In <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070725#pone-0070725-g004" target="_blank">Fig. 4 A, B</a>, GFP expression and the number of GFP-positive cells in miR-140-treated cultures were much lower than those in D-NC- or ASO-140-treated controls. C: Western blot analysis. The gene expression/α-tubulin ratios were shown on the right of gel. The miR-140 treatment led to an obvious reduction in the expression of TGF-β1, vimentin, Smad3 and p-Smad3 compared to the D-NC control groups. α-tubulin was used as loading controls. *<i>P</i><0.05 versus miR-140 treatment, **<i>P</i><0.05 versus ASO-140 treatment, n = 3 replicates.</p

PTX ameliorates pulmonary fibrosis andu pregulates miR-140 in fibrotic lungs.

<p>A: Upper, Haematoxylin and eosin (HE). The lungs initiated alveolar disruption on 7 d, and more severe on 28 d, while pulmonary fibrosis was ameliorated by PTX (0.6 mg/kg) treatment. Lower, Masson's trichrome for collagen and elastin evaluation. The lungs underwent a slight fibrotic invasion on 7 d, and more aggressive on 28 d, which was alleviated by PTX. Scale bars: 150 µm. B: Type-I collagen ELISA analysis. The type-I collagen level showed basally low in lung tissues of rats instilled with saline (sham control rats), similar to those in the untreated lungs (0 d). The collagen I levels in BLM-treated lung tissues on 28 d were increased seven folds than those in untreated or sham control lung tissues. Collagen I levels were reduced obviously in PTX-treated lungs compared to BLM-treated 28 d lung tissues. *<i>P</i><0.05 versus 28 d. C: Hydroxyproline analysis. BLM-treated lungs had much higher levels of hydroxyproline, but PTX treatment significantly decreased hydroxyproline levels compared to BLM-treated 28 d lungs. *<i>P</i><0.05 versus 28 d. D: Quantitative RT-PCR analysis of miR-140 expression. U6 was used as a control. n = 3 replicates. The miR-140 level in BLM-instilled rat lungs reached its nadir on 7 d, approximately two folds lower than that in untreated or sham control lung tissues, while it was dramatically upregulated in PTX-treated lung tissues. *<i>P</i><0.05 versus 0 d. **<i>P</i><0.05 versus PTX. E: miR-140 expression in human pulmonary fibrotic lungs. a–e: healthy control lungs, f–j: human pulmonary fibrosis lungs. The miR-140 levels are obvious lower in humanulmonary fibrotic lung tissues compared with those in healthy control lungs.</p

MiR-140 affected EMT and Smad3/p-Smad3 expression.

<p>A: Morphological characteristics. Scale bars: 150 µm. A549 cells treated with TGF-β1 or TGF-β1+ D-NC underwent a fibroblast-like morphology, while their morphologies were reversed by miR-140 or miR-140 plus PTX treatment. B: Western blot analysis. α-tubulin was used as loading controls. The Smad3 and p-Smad3 levels were decreased obviously in miR-140- or PTX+miR-140-treated A549 cells, especially in PTX+miR-140-treated cells, compared to those in TGF-β1- or TGF-β1+D-NC-treated cultures. n = 3 replicates.</p

PTX suppresses the TGF-β1/Smad3 signaling pathway.

<p>A: Smad3 and p-Smad3 expression in A549 cells by western blot analysis. α-tubulin was used as loading controls. B: Quantitative RT-PCR analysis of Smad3 expression in A549 cells. GAPDH was used a control gene. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070725#pone-0070725-g003" target="_blank">In Fig. 3A,B</a>, TGF-β1-treatment increased the expression of p-Smad3 and Smad3 in A549 cells, which could be reduced by PTX treatment. *<i>P</i><0.05 versus TGF-β1 treatment group, n = 3 replicates. C: Immunohistochemical staining of Smad3, p-Smad3 and α-SMA in rat lung tissues. Black arrows, Smad3, p-Smad3 or α-SMA positive staining AECs. Stars, vasculature. Scale bars: 150 µm. A majority of Smad3, p-Smad3 and α-SMA staining was found to be localized in the AECs of fibrotic lungs. The BLM-untreated (0 d) or sham control lung tissues showed almost a total absence of α-SMA and p-Smad3. TGF-β1-treatment increased the expression of p-Smad3 and α-SMA in fibrotic lungs (7 d, 28 d), which was restored by PTX treatment to some extent. D: Western blot analysis. E: The ratios of Smad3 (p-Smad3, or α-SMA)/α-tubulin of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070725#pone-0070725-g003" target="_blank">Fig. 3D</a>. α-tubulin was used as the internal standard. n = 3 replicates. F: Quantitative RT-PCR analysis of Smad3 and α-SMA expressions in rat lung tissues (n = 3 replicates). GAPDH was used as a control gene. In <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070725#pone-0070725-g003" target="_blank">Fig. 3D–F</a>: the quantity of p-Smad3 in lung tissues reached peaked on day 7 and the quantity of α-SMA in lung tissues reached peaked on day 28 after BLM treatment. PTX could reduce the expression of p-Smad3 and α-SMA obviously, but had no effect on Smad3. *<i>P</i><0.05 versus 0 d. **<i>P</i><0.05 versus PTX.</p

Lung-to-body weight ratio, the pulmonary inflammation and fibrosis scores.

*<p><i>P</i><0.05 versus saline-treated control,</p>**<p><i>P</i><0.05 versus BLM-treated 28 d rats, n = 12.</p

Increased Expression of Pregnancy Up-Regulated Non-Ubiquitous Calmodulin Kinase Is Associated with Poor Prognosis in Clear Cell Renal Cell Carcinoma

Purpose: The aims of this study were to evaluate the clinical significance and potential prognostic value of pregnancy up-regulated non-ubiquitous calmodulin kinase (PNCK) in clear cell renal cell carcinoma (ccRCC) patients. Materials and Methods: The expression of PNCK mRNA was determined in 24 paired samples of ccRCCs and adjacent normal tissues using real-time RT-PCR. The expression of PNCK was determined in 248 samples of ccRCCs and 92 paired samples of adjacent normal tissues by immunohistochemical analysis. Statistical analysis was performed to define the relationship between PNCK expression and the clinical features of ccRCC. Results: The mRNA level of PNCK was significantly higher in tumorous tissues than in the adjacent non-tumorous tissues (p<0.001). An immunohistochemical analysis of 92 paired tissue specimens showed that PNCK expression was higher in tumorous tissues than in the adjacent non-tumorous tissues (p<0.001). Moreover, there was a significant correlation between the PNCK expression and various clinicopathological parameters such as Fuhrman grade (p = 0.011), tumor size (p<0.001), T stage (p<0.001) and N stage (p = 0.015). Patients with higher PNCK expression had shorter overall survival time than those with lower PNCK expression (p<0.001). Multivariate analysis indicated that PNCK expression was an independent predictor for poor survival of ccRCC patients. Conclusions: To our knowledge, this is the first study that determines the relationship between PNCK and prognosis in ccRCC. We found that increased PNCK expression is associated with poor prognosis in ccRCC. PNCK may represent a novel prognostic marker for ccRCC