Atorvastatin Upregulates the Expression of miR-126 in Apolipoprotein E-knockout Mice with Carotid Atherosclerotic Plaque

Carotid atherosclerosis (AS) is a chronic inflammatory disease of the carotid arterial wall, which is very important in terms of the occurrence of cerebral vascular accidents. Studies have demonstrated that microRNAs (miRNAs) and their target genes are involved in the formation of atherosclerosis and that atorvastatin might reduce atherosclerotic plaques by regulating the expression of miRNAs. However, the related mechanism is not yet known. In this study, we first investigated the effects of atorvastatin on miR-126 and its target gene, i.e., vascular cell adhesion molecule-1 (VCAM-1) in apolipoprotein E-knockout (ApoE?/?) mice with carotid atherosclerotic plaque in vivo. We compared the expressions of miR-126 and VCAM-1 between the control, atherosclerotic model and atorvastatin treatment groups of ApoE?/? mice using RT-PCR and Western blot. We found the miR-126 expression was significantly down-regulated, and the VCAM-1 expression was significantly up-regulated in the atherosclerotic model group, which accelerated the progression of atherosclerosis in the ApoE?/? mice. These results following atorvastatin treatment indicated that miR-126 expression was significantly up-regulated, VCAM-1 expression was significantly down-regulated and atherosclerotic lesions were reduced. The present results might explain the mechanism by which miR-126 is involved in the formation of atherosclerosis in vivo. Our study first indicated that atorvastatin might exert its anti-inflammatory effects in atherosclerosis by regulating the expressions of miR-126 and VCAM-1 in vivo.     

Vimentin is important in the neural differentiation of PC12 cells promoted by sialylation

Sialic acid modification is a kind of post-translational modification. To investigate the regulation effect of sialic acid on neural differentiation, we used CycloManN propanyl perac (CycloManN pro), a metabolic precursor of sialic acid, to treat PC12 cells. We noted that CycloManN pro indeed robustly promoted global sialylation detected by MAL II lectin blot in PC12 cells. Simultaneously, we interestingly found that the neurite outgrowth of PC12 cells was significantly promoted by the CycloManN pro treatment. The profile analysis of sialylated proteins showed that a protein band at 55KD was greatly enhanced especially in PC12L cells after CycloManN pro treatment. After enrichment with lectin MAL II, the proteins in this band were analyzed by mass spectrometry. The results showed that 23 proteins were in the band, but the score of vimentin was the highest among them. To investigate further the role of vimentin in the process of neurite differentiation, vimentin construct was transfected into PC12 cells. We interestingly observed that ectopic expression of vimentin significantly enhanced the neurite outgrowth induced by CycloManN pro. However, after three potential glycosylation sites (Ser-7, Thr-33, Ser-34:) of vimentin were mutated to alanine, overexpression of the mutated vimentin completely lost the enhancement activity for the neural differentiation even in the presence of CycloManN pro. Taken together, our study demonstrated that vimentin was important in the induction of neural differentiation by CycloManN pro.     

Large numbers of interleukins‐22‐ and ‐17A‐producing T helper cells in cholangiocarcinoma related to liver fluke infection

Cholangiocarcinoma (CCA) associated with liver fluke infection involves inflammatory and immune processes; however, whether these involve the proinflammatory cytokine IL‐17A and proliferative cytokine IL‐22 remains unclear. Here, numbers of IL‐22‐ and IL‐17A‐producing Th cells and cytokine concentrations in 30 patients with CCA and long‐term liver fluke infection, 40 patients with liver‐fluke infection but not CCA, and 16 healthy controls were compared. Analyses were performed using immunohistochemistry, flow cytometry, ELISA and RT‐PCR. Immunohistochemical staining showed weaker expression of IL‐22 and IL‐17A in patients with CCA with than in those without liver fluke infection (P < 0.01). Flow cytometry revealed significantly greater median proportions of IL‐22‐producing T helper cells in patients with CCA (2.2%) than in those without it (0.69%) or controls (0.4%, P < 0.001). Similar results were obtained for IL‐17A‐producing T helper cells. ELISA revealed plasma concentrations of IL‐22 were 1.3‐fold higher in patients with CCA than in those without it and 4.6‐fold higher than in controls (P < 0.001). Plasma concentrations of IL‐17A were 2.5‐fold higher in patients with CCA than in those without it, and 21‐fold higher than in controls (P < 0.001). Amounts of IL‐22 and IL‐17A mRNAs in blood were significantly higher in patients with CCA than in the other two groups. Proportions of CD4⁺CD45RO⁺ T cells producing IL‐22 correlated with proportions producing IL‐17A (r = 0.759; P < 0.001), and plasma concentrations of IL‐22 correlated with those of IL‐17A (r = 0.726; P < 0.001). These results suggest that both IL‐17A and IL‐22 affect development of CCA related to liver fluke infection.

     

偃麦草属植物醇溶蛋白和谷蛋白多态性及系统学研究

运用酸性聚丙烯酰胺凝胶电泳(A-PAGE)和十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)对偃麦草属(Elytrigia)24个物种的醇溶蛋白和谷蛋白进行了研究。以中国春为参照,按醇溶蛋白和谷蛋白电泳图谱条带迁移距离大小和条带多态性对供试材料进行聚类分析。结果显示,24份供试材料均呈现不同的醇溶蛋白和谷蛋白电泳图谱,共分离出醇溶蛋白带纹83条和谷蛋白带纹53条,多态性均达100%,并且在相同染色体组组成的物种中,染色体数目越多,其蛋白带纹越多;偃麦草属24个物种的醇溶蛋白和谷蛋白图谱均有明显差异,蛋白图谱可作为鉴定偃麦草属物种的指纹图谱。聚类分析结果显示,在材料间的醇溶蛋白遗传相似性系数为0.66时,24份材料被划分为6个主要类群,含E和St基因组的物种具有较近的亲缘关系;在谷蛋白遗传相似性系数为0.62时,24份材料被聚为4大类。聚类结果表明,染色体组成未知的物种Et.pachynera可能为异源多倍体物种。     

Targeting myeloid differentiation protein 2 by the new chalcone L2H21 protects LPS‐induced acute lung injury

Acute inflammatory diseases are the leading causes of mortality in intensive care units. Myeloid differentiation 2 (MD‐2) is required for recognizing lipopolysaccharide (LPS) by toll‐like receptor 4 (TLR4), and represents an attractive therapeutic target for LPS‐induced inflammatory diseases. In this study, we report a chalcone derivative, L2H21, as a new MD2 inhibitor, which could inhibit LPS‐induced inflammation both in vitro and in vivo. We identify that L2H21 as a direct inhibitor of MD‐2 by binding to Arg⁹⁰ and Tyr¹⁰² residues in MD‐2 hydrophobic pocket using a series of biochemical experiments, including surface plasmon response, molecular docking and amino acid mutation. L2H21 dose dependently inhibited LPS‐induced inflammatory cytokine expression in primary macrophages. In mice with LPS intratracheal instillation, L2H21 significantly decreased LPS‐induced pulmonary oedema, pathological changes in lung tissue, protein concentration increase in bronchoalveolar lavage fluid, inflammatory cells infiltration and inflammatory gene expression, accompanied with the decrease in pulmonary TLR4/MD‐2 complex. Meanwhile, administration with L2H21 protects mice from LPS‐induced mortality at a degree of 100%. Taken together, this study identifies a new MD2 inhibitor L2H21 as a promising candidate for the treatment of acute lung injury (ALI) and sepsis, and validates that inhibition of MD‐2 is a potential therapeutic strategy for ALI.     

P2X7 receptor inhibition attenuated sympathetic nerve sprouting after myocardial infarction via the NLRP3/IL‐1β pathway

Mounting evidence supports the hypothesis that inflammation modulates sympathetic sprouting after myocardial infarction (MI). The myeloid P2X₇ signal has been shown to activate the nucleotide‐binding and oligomerization domain‐like receptor family pyrin domain‐containing 3 (NLRP3) inflammasome, a master regulator of inflammation. We investigated whether P2X₇ signal participated in the pathogenesis of sympathetic reinnervation after MI, and whether NLRP3/interleukin‐1β (IL‐1β) axis is involved in the process. We explored the relationship between P2X₇ receptor (P2X₇R) and IL‐1β in the heart tissue of lipopolysaccharide (LPS)‐primed naive rats. 3′‐O‐(4‐benzoyl) benzoyl adenosine 5′‐triphosphate (BzATP), a P2X₇R agonist, induced caspase‐1 activation and mature IL‐1β release, which was further neutralized by a NLRP3 inhibitor (16673‐34‐0). MI was induced by coronary artery ligation. Following infarction, a marked increase in P2X₇R was localized within infiltrated macrophages and observed in parallel with an up‐regulation of NLRP3 inflammasome levels and the release of IL‐1β in the left ventricle. The administration of A‐740003 (a P2X₇R antagonist) significantly prevented the NLRP3/IL‐1β increase. A‐740003 and/or Anakinra (an IL‐1 receptor antagonist) significantly reduced macrophage infiltration as well as macrophage‐based IL‐1β and NGF (nerve growth factor) production and eventually blunted sympathetic hyperinnervation, as assessed by the immunofluorescence of tyrosine hydroxylase (TH) and growth‐associated protein 43 (GAP 43). Moreover, the use of Anakinra partly attenuated sympathetic sprouting. This indicated that the effect of P2X₇ on neural remodelling was mediated at least partially by IL‐1β. The arrhythmia score of programmed electric stimulation was in accordance with the degree of sympathetic hyperinnervation. In vitro studies showed that BzATP up‐regulated secretion of nerve growth factor (NGF) in M1 macrophages via IL‐1β. Together, these data indicate that P2X₇R contributes to neural and cardiac remodelling, at least partly mediated by NLRP3/IL‐1β axis. Therapeutic interventions targeting P2X₇ signal may be a novel approach to ameliorate arrhythmia following MI.     

VEGF165 induces differentiation of hair follicle stem cells into endothelial cells and plays a role in in vivo angiogenesis

Within the vascular endothelial growth factor (VEGF) family of five subtypes, VEGF165 secreted by endothelial cells has been identified to be the most active and widely distributed factor that plays a vital role in courses of angiogenesis, vascularization and mesenchymal cell differentiation. Hair follicle stem cells (HFSCs) can be harvested from the bulge region of the outer root sheath of the hair follicle and are adult stem cells that have multi‐directional differentiation potential. Although the research on differentiation of stem cells (such as fat stem cells and bone marrow mesenchymal stem cells) to the endothelial cells has been extensive, but the various mechanisms and functional forms are unclear. In particular, study on HFSCs’ directional differentiation into vascular endothelial cells using VEGF165 has not been reported. In this study, VEGF165 was used as induction factor to induce the differentiation from HFSCs into vascular endothelial cells, and the results showed that Notch signalling pathway might affect the differentiation efficiency of vascular endothelial cells. In addition, the in vivo transplantation experiment provided that HFSCs could promote angiogenesis, and the main function is to accelerate host‐derived neovascularization. Therefore, HFSCs could be considered as an ideal cell source for vascular tissue engineering and cell transplantation in the treatment of ischaemic diseases.     

Stable Inverted Planar Perovskite Solar Cells with Low‐Temperature‐Processed Hole‐Transport Bilayer

Low‐temperature‐processed perovskite solar cells (PSCs), which can be fabricated on rigid or flexible substrates, are attracting increasing attention because they have a wide range of potential applications. In this study, the stability of reduced graphene oxide and the ability of a poly(triarylamine) underlayer to improve the quality of overlying perovskite films to construct hole‐transport bilayer by means of a low‐temperature method are taken advantage of. The bilayer is used in both flexible and rigid inverted planar PSCs with the following configuration: substrate/indium tin oxide/reduced graphene oxide/polytriarylamine/CH₃NH₃PbI₃/PCBM/bathocuproine/Ag (PCBM = [6,6]‐phenyl‐C₆₁‐butyric acid methyl ester). The flexible and rigid PSCs show power conversion efficiencies of 15.7 and 17.2%, respectively, for the aperture area of 1.02 cm². Moreover, the PSC based the bilayer shows outstanding light‐soaking stability, retaining ≈90% of its original efficiency after continuous illumination for 500 h at 100 mW cm^?2.     

Four New Diterpene Glucosides from Perovskia atriplicifolia

Four new diterpene glucosides, namely perovskiaditerpenosides A – D ( 1  –  4 ), were isolated from the BuOH extract of Perovskia atriplicifolia. Their structures were well elucidated by chemical methods and comprehensive spectroscopic analyses including MS, IR, and NMR (1D and 2D). The newly isolated compounds were screened for their cytotoxic activity against HepG2, NB4, HeLa, K562, MCF7, PC3, and HL60. The obtained results indicated that the new compounds possessed considerable cytotoxic activity.     

Cytotoxic and Anti‐inflammatory Sesquiterpenes from Ainsliaea henryi

Three new sesquiterpenoids, 4α‐hydroxyeudesm‐11(13)‐en‐12‐yl 3‐methylbutanoate ( 1 ), diaspanolide E ( 2 ), and (13α)‐germacra‐1(10),4‐dien‐12,8α‐olid‐15‐oic acid ( 3 ), along with eight known sesquiterpenoids ( 4 – 11 ), were isolated from the aerial parts of Ainsliaea henryi. The chemical structures of compounds 1 – 3 were elucidated by spectroscopic analysis (1D‐, 2D‐NMR, MS and HR/MS). All isolates were evaluated for their inhibitory activities against nitric oxide (NO) production in lipopolysaccharide‐induced RAW264.7 macrophage cells. Compound 10 exhibited significantly inhibition against NO release with an IC₅₀ value of 6.54 ± 0.16 μm . Also, all isolated compounds were tested for cytotoxicity against three human tumor cell lines A549, MGC803, and HCT116, among which compound 5 significantly inhibited the proliferation of MGC803 cell lines with an IC₅₀ value of 2.2 ± 0.2 μm .