Neutrophil extracellular traps (NETs) facilitate the extracellular killing of pathogens. However, excessive NETs formation and poor degradation are associated with exacerbated immune responses and tissue injury. In this study, we investigated the role of NETs in lipopolysaccharide (LPS)-mediated acute lung injury (ALI) and assessed the use of DNase I, for the treatment of ALI. Additionally, we focused on the controversial issue of whether LPS directly induces NETs release in vitro. NETs formation was detected in murine ALI tissue in vivo and was associated with increased NETs markers, citrullinated-histone H3 tissue levels and NET-DNA levels in BALF. Treatment with DNase I significantly degraded NETs and reduced citrullinated-histone H3 levels, which protected against ALI and ameliorated pulmonary oedema and total protein in BALF. In addition, DNase I significantly reduced IL-6 and TNF-α levels in plasma and BALF. In vitro, LPS-activated platelets rather than LPS alone efficiently induced NETs release. In conclusion, NETs formed during LPS-induced ALI, caused organ damage and initiated the inflammatory response. NETs degradation by DNase I promoted NET-protein clearance and protected against ALI in mice; thus, DNase I may be a new potential adjuvant for ALI therapy. Specifically, LPS induced NETs formation in an indirect manner via platelets activation.

BackgroundThe main approach to treat HIV-1 infection is combination antiretroviral therapy (cART). Although cART is effective in reducing HIV-1 viral load and controlling disease progression, it has many side effects, and is expensive for HIV-1 infected patients who must remain on lifetime treatment. HIV-1 gene therapy has drawn much attention as studies of genome editing tools have progressed. For example, zinc finger nucleases (ZFN), transcription activator like effector nucleases (TALEN) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 have been utilized to successfully disrupt the HIV-1 co-receptors CCR5 or CXCR4, thereby restricting HIV-1 infection. However, the effects of simultaneous genome editing of CXCR4 and CCR5 by CRISPR-Cas9 in blocking HIV-1 infection in primary CD4+ T cells has been rarely reported. Furthermore, combination of different target sites of CXCR4 and CCR5 for disruption also need investigation.ResultsIn this report, we designed two different gRNA combinations targeting both CXCR4 and CCR5, in a single vector. The CRISPR-sgRNAs-Cas9 could successfully induce editing of CXCR4 and CCR5 genes in various cell lines and primary CD4+ T cells. Using HIV-1 challenge assays, we demonstrated that CXCR4-tropic or CCR5-tropic HIV-1 infections were significantly reduced in CXCR4- and CCR5-modified cells, and the modified cells exhibited a selective advantage over unmodified cells during HIV-1 infection. The off-target analysis showed that no non-specific editing was identified in all predicted sites. In addition, apoptosis assays indicated that simultaneous disruption of CXCR4 and CCR5 in primary CD4+ T cells by CRISPR-Cas9 had no obvious cytotoxic effects on cell viability.ConclusionsOur results suggest that simultaneous genome editing of CXCR4 and CCR5 by CRISPR-Cas9 can potentially provide an effective and safe strategy towards a functional cure for HIV-1 infection.Electronic supplementary materialThe online version of this article (doi:10.1186/s13578-017-0174-2) contains supplementary material, which is available to authorized users.

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Solution‐processed lead halide perovskites have shown good applicability in both solar cells and microlasers. Very recently, the nonlinear properties of perovskites have attracted considerable research attention. Second harmonic generation and two‐photon absorption have been successfully demonstrated. However, perovskite devices based on these nonlinear properties, such as micro‐ and nanolasers have thus far not been fabricated. Here we demonstrate two‐photon pumped microlasers from CH3NH3PbBr3 perovskite microwires. These CH3NH3PbBr3 perovskite microwires are synthesized through a one‐step solution precipitation method and dispersed on a glass substrate. Under optical excitation at 800 nm, two‐photon pumped lasing action with periodic peaks is successfully observed at around 546 nm. The obtained quality (Q) factors of the two‐photon pumped microlasers are around 682, and the corresponding thresholds are about 674 µJ cm‐2. Both the Q factors and thresholds are comparable to conventional whispering‐gallery modes in two‐dimensional polygon microplates. This work is the first demonstration of two‐photon pumped microlasers in CH3NH3PbBr3 perovskite microwires. We believe our finding will significantly expand the application of perovskites in low‐cost nonlinear optical devices, such as optical limiters, optical switches, and biomedical imaging devices.