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Elabela (ELA), which is the second endogenous peptide ligand of the apelin receptor (APJ) to be discovered, has been widely studied for potential use as a therapeutic peptide. However, its role in ischemic stroke (IS), which is a leading cause of disability and death worldwide and has limited therapeutic options, is uncertain. The aim of the present study was to investigate the beneficial effects of ELA on neuron survival after ischemia and the underlying molecular mechanisms. Primary cortical neurons were isolated from the cerebral cortex of pregnant C57BL/6J mice. Flow cytometry and immunofluorescence showed that ELA inhibited oxygen–glucose deprivation (OGD) -induced apoptosis and axonal damage in vitro. Additionally, analysis of the Gene Expression Omnibus database revealed that the expression of microRNA-124-3p (miR-124-3p) was decreased in blood samples from patients with IS, while the expression of C-terminal domain small phosphatase 1 (CTDSP1) was increased. These results indicated that miR-124-3p and CTDSP1 were related to ischemic stroke, and there might be a negative regulatory relationship between them. Then, we found that ELA significantly elevated miR-124-3p expression, suppressed CTDSP1 expression, and increased p-AKT expression by binding to the APJ receptor under OGD in vitro. A dual-luciferase reporter assay confirmed that CTDSP1 was a direct target of miR-124-3p. Furthermore, adenovirus-mediated overexpression of CTDSP1 exacerbated neuronal apoptosis and axonal damage and suppressed AKT phosphorylation, while treatment with ELA or miR-124-3p mimics reversed these effects. In conclusion, these results indicated that ELA could alleviate neuronal apoptosis and axonal damage by upregulating miR-124-3p and activating the CTDSP1/AKT signaling pathway. This study, for the first time, verified the protective effect of ELA against neuronal injury after ischemia and revealed the underlying mechanisms. We demonstrated the potential for the use of ELA as a therapeutic agent in the treatment of ischemic stroke. Graphical Abstract

The "Supplemental Data" contains supplementary methods,supplementary table 1 and supplementary figures1-19.1.Supplementary methods: Antibodies and Reagents, Primary culture of LECs, Immunomagnetic isolation of LECs, Expressing human recombinant sSEMA4C protein, Liquid chromatography - Electrospray ionization tandem MS (LC-ESI/MS), Migration assay, Wound healing assay, Cell proliferation assay, Primers sequences; 2.Supplementary Table 1: the Patient characteristics; 3.Supplementary Figure 1: the Authentication of LECs; 4.Supplementary Figure 2: the Evaluation of LCM-isolated LECs using RNA markers; 5. Supplementary Figure 3 : SEMA4C expresses on lymphatic vessels; 6.Supplementary Figure 4 : Nano-LC-ESI-MS/MS spectrum of a single charged deglycosylated glycopeptide from the culture supernatants of tumor LECs; 7. Supplementary Figure 5 :sSEMA4C binds to tumor cells and LECs; 8.Supplementary Figure 6: sSEMA4C promotes the migration and proliferation of tumor cells; 9.Supplementary Figure 7: SEMA4C promotes LECs function in vitro; 10.Supplementary Figure 8 : targeting SEMA4C inhibits LECs function in vitro; 11.Supplementary Figure 9 : the growth of the tumors; 12.Supplementary Figure 10 : Knock-down of PlexinBs; 13.Supplementary Figure 11 : Blocking ERBB2 or PlexinB2 inhibits SEMA4C-induced lymphangiogenesis in vitro; 14.Supplementary Figure 12 : LEC was stimulated by VEGF-C and sSEMA4C; 15.Supplementary Figure 13 : sSEMA4C activates MET and RHOA in tumor cells; 16.Supplementary Figure 14 : blocking MET or PlexinB2 inhibits SEMA4C-induced migration and proliferation of tumor cells; 17.Supplementary Figure 15 : cells treated with sSEMA4C; 18.Supplementary Figure 16 : targeting RHOA inhibits SEMA4C-induced lymphangiogenesis in vitro and in vivo; 19.Supplementary Figure 17 : targeting RHOA inhibits SEMA4C-induced migration of tumor cells; 20.Supplementary Figure 18 : 3 days after inoculation with tumour cells, the mice were treated with Lapatinib, K252a ,C3 toxin, Fasudil, Y27632 , sSEMA4C and PBS control solution, the size of tumor was measured; 21.Supplementary Figure 19: 3 days after inoculation with tumour cells, the mice were treated with Lapatinib, K252a ,C3 toxin, Fasudil, Y27632 , sSEMA4C and PBS control solution, the average number of positive lymph node in each mice were calculated.

Purpose: Lymphatic vessels are mainly regarded as passive conduits for the dissemination of cancer cells. In this study, we investigate whether and how the tumor-associated lymphatic vessels may play an active role in tumor metastasis.Experimental Design:In situ laser capture microdissection of lymphatic vessels followed by cDNA microarray analysis was used to determine the expression profiling of lymphatic endothelial cells (LEC). Gene expression levels and activity of signaling pathways were measured by real-time RT-PCR, ELISA, or immunoblotting. Lymphangiogenesis was assessed by IHC. Lymph node metastasis was measured using fluorescence imaging. The effects of SEMA4C on lymphangiogenesis in vitro were evaluated using migration assay and tube-formation assay of LECs.Results: Tumor-associated LECs are molecularly and functionally different from their normal counterparts. In addition to expressing high levels of membrane-bound SEMA4C, tumor-associated LECs also produced soluble SEMA4C (sSEMA4C). Increased serum sSEMA4C was detected in patients with breast cancer and cervical cancer. Patients with metastasis had much higher levels of serum sSEMA4C. sSEMA4C promoted lymphangiogenesis by activating PlexinB2-ERBB2 signaling in LECs, and promoted the proliferation and migration of tumor cells by activating PlexinB2-MET signaling, thus promoting lymphatic metastasis. Although the SEMA4C signaling pathways differ between LECs and tumor cells, RHOA activation was necessary for the effects of SEMA4C in both types of cells.Conclusions: Tumor-associated LECs produce sSEMA4C to promote lymphatic metastasis of tumors. Our results suggest that SEMA4C and RHOA might be potential therapeutic targets, and that higher serum sSEMA4C could be a marker for breast cancer and cervical cancer. Clin Cancer Res; 23(1); 214–24. ©2016 AACR.

Flavonoids are rich in tea plants (Camellia sinensis), and responsible for the flavor and healthful benefits of tea beverage. The anthocyanin levels in the purple tender shoots are higher than in the general green leaves of tea plant, which provide special materials to search metabolic mechanisms of flavonoid enrichment in plant. In this work, flavonoid differences between purple and green shoots from tea cultivars “Zijuan” (ZJ) and “Yunkang10” (YK-10) were investigated through metabolomic analysis, and mechanisms for their difference were surveyed by comparative transcriptomic and proteomic analysis. Levels of 34 flavonoids were different between ZJ and YK-10 shoots. Among them, 8 and 6 were marker metabolites in ZJ and YK-10, respectively. The differentially expressed genes (DEGs), differentially expressed proteins (DEPs), and different-level metabolites (DLMs) between ZJ and YK-10 were researched, respectively; and interactions including DEG-DLM, DEP-DLM, DEG-DEP, and DEG-DEP-DLM were analyzed; the contents of 18 characteristic flavonoids in tea leaves and expressions of 34 flavonoid metabolic genes were measured to verify the omics results. Integrated above analyses, a proposed model of flavonoids biosynthesis in tea shoots were established. The differential expression of the leucoanthocyanidin reductase (LAR), anthocyanidin synthase (ANS), anthocyanidin reductase (ANR), UDPG-flavonoid glucosyltransferase (UGT) 75L12 and 94P1 at gene level, and the ANS, ANR, and UGT78A15 at protein level, were closely associated with differences in flavonoids between ZJ and YK-10 shoot. Together, this study provides new information on the flavonoid accumulation mechanism in tea plant.

Pathological hypoxia/oxidative stress of placenta is a major cause in preeclampsia. Autophagy dysregulation has recently been linked to aberrant placental vascular function in preeclampsia [1, 2] while the underlying mechanisms remain unknown. Here, we found exosomal TINCR derived from trophoblasts exposed to hypoxia-oxidative stress induced placental vascular dysfunction through evoking excessive autophagic activation of endothelial cells related to preeclampsia. Specifically, umbilical cord serum- and plasma-derived exosomal TINCR levels were elevated in preeclampsia determined by microarray-based transcript analyses and qRT-PCR, which may be caused by trophoblasts releasing exosomes containing TINCR into circulation under pathological hypoxia conditions of the placenta. These exosomal TINCR impaired placental vascular function by promoting excessive autophagy of endothelial cells involved in preeclampsia-like symptoms both in vivo and in vitro. In contrast, the autophagy inhibitor hydroxychloroquine (HCQ) partially reversed exosomal TINCR's effect on placental vascular dysfunction and preeclampsia symptoms. Mechanistically, TINCR promoted autophagy in endothelial cells by regulating miR-424/ ATG5 axis as a competing endogenous RNA. Therefore, preeclampsia was associated with the overactivation of endothelial cell autophagy by trophoblast-derived exosomal TINCR in regulating placental vascular function, indicating that targeting autophagy such as HCQ is a novel strategy for preeclampsia treatment.

Background:This study aims to compare the histological and thickness changes of attached gingiva following grafting with three groups of different soft tissue substitutes at labial aspect of canines. Methods: In three beagle dogs, attached gingival thickening was performed with double layer allogeneic acellular dermal matrix membrane (AADM), bovine acellular dermal matrices membrane (BADM) combined with concentrated growth factor membrane (CGF) or BADM combined with collagen sponge. Thickness was measured before augmentation (baseline [BL]), and after 1, 2, 3, and 4 months. Descriptive histological analyses were performed after 4 months.Results: Four months post-operatively, thickness increase differed significantly among three groups (AADM: 1.88±0.28 mm; BADM combined with CGF: 0.27±0.19 mm; BADM combined with collagen sponge: 0.10±0.11 mm). Compared to BL, AADM group showed significant difference (3.24±0.22 mm vs. 1.36±0.72 mm), while the BADM combined with CGF group showed no significant difference (1.50±0.07 mm vs. 1.23±0.15 mm) and neither of the BADM combined with collagen sponge group (1.54±0.15 mm vs. 1.44±0.15 mm). Histological analyses revealed that AADM was integrated with gingival tissues, and new fibrous tissue and blood vessels were formed. But the grafts in the other two groups were absorbed and the augmentation area was remodeled into a structure consistent with the surrounding host tissue.Conclusions: . AADM is superior to BADM combined with CGF and BADM combined with collagen sponge with regard to gingival augmentation.

Objective Human immunodeficiency virus type 1 (HIV-1) infection disturbs the balance of CD4+ T cells and monocytes in the immune system. In the early stage of infection, the virus stimulates the activation and proliferation of immune cells, induces the release of cytokines, destroys CD4+ T cells, and accelerates HIV-1 replication and AIDS progression. It is essential to explore cytokine changes after HIV-1 infection and further understand the underlying mechanism of HIV infection. Materials and methods In this study, we enrolled 38 HIV-infected subjects and 30 healthy subjects. We measured and compared CD4+ T cell counts, the tropism of HIV and the serum cytokine levels in different groups. Results Our results showed significantly higher serum levels of IL-1β, IL-2, IL-4, IL-7, IL-10, IL-17, IFN-γ, and TNF-α in HIV-infected patients. Higher levels of IL-6 and IL-17 were observed in the + T cell count group, and higher levels of IL-2 were observed in the CCR5-tropic HIV strain group. Conclusion In conclusion, we found that HIV infection-induced activation of the immune system. IL-6 and IL-17 may predict the severity of HIV disease and regulate HIV infection. The level of IL-2 depended on the type of virus strain.

Surface defects on optical fiber end-face, such as scratches, cause heavy loss and low data-transfer rate. Therefore, the inspection of optical fiber end-face defect cannot be neglected. Most optical fiber defect detection methods ignore the striped pattern of scratches and easily cause incomplete segmentation. In this article, a novel method based on deep convolutional neural networks (DCNNs) for weak microscratches inspection in the optical fiber end-face is proposed. First, a new feature pyramid structure with criss-cross max-pooling (CCMP) block is developed to extract precise boundary features in all directions and generate feature maps. Second, these feature maps are fed into the spatial attention upsample (SAU) module to recover a full-resolution feature map and localize the defects. Finally, multiple scratches with various types can be detected in the images. This article uses many real optical fiber images as experimental images. The experimental results demonstrate that the proposed method yields good comprehensive performance with a mean pixel accuracy (mPA) of 92.70% and a mean intersection over union (mIoU) of 85.35%.

ObjectiveWe evaluated and compared the efficacy and safety of neoadjuvant chemoradiotherapy (NACRT) versus neoadjuvant chemotherapy (NACT) for locally advanced gastric cancer (LAGC) in a single-center randomized phase II trial.MethodsPatients with LAGC were enrolled and received either NACT or NACRT, followed by gastrectomy and adjuvant chemotherapy. The primary endpoint was an R0 resection rate.ResultsWe enrolled 75 patients: 75.7% (NACT, 28/37 patients) and 76.3% (NACRT, 29/38 patients) underwent surgery; R0 resection rates were 73.0% (27/37) and 73.7% (28/38), respectively. The NACRT group had significantly better major pathological response than the NACT group (37.9% vs 17.9%, p = 0.019). Between-group postoperative complications were not significantly different. The median follow-up was 59.6 months; 5-year overall survival (OS) rate was 50.1% (NACT) and 61.9% (NACRT); neither group reached the median OS; median progression-free survival was 37.3 and 63.4 months, respectively.ConclusionsS-1-based NACRT did not improve the R0 resection rate, although it presented better tumor regression with similar safety to NACT.Trial registrationClinicalTrial.gov NCT02301481

Semiconductor wafer is widely used in welding robot, spray robot, unmanned material delivery vehicle, and detection station sensor. The defects of semiconductor wafer, such as stains, burrs, scratches, and holes generated in the manufacturing process, severely affect the quality of downstream products. Therefore, the inspection of wafer defect could not be neglected. Traditional semiconductor wafer defect inspection methods based on handcrafted features heavily rely on the expertise and are limited in some application scenario. In this article, a novel method based on deep convolutional neural networks for semiconductor wafer surface defect inspection is proposed. First, a new structure of feature pyramid networks with atrous convolution (FPNAC) is developed to extract the features and to generate feature maps. Second, the feature maps are fed into region proposal network (RPN) to generate region proposals. Finally, the region proposals are aligned to corresponding size as the inputs of deep multibranches neural network (DMBNN) consisting of three branches, to classify and segment the defects precisely. Experimental results demonstrate that the proposed method yields good comprehensive performance with mean pixel accuracy (MPA) 93.97% and mean intersection over union (MIoU) 83.58%.

Herein, a binuclear ruthenium(II) complex modified by calix[4]arene group has been prepared. The complex as potential inducer and stabilizer of c-myc G-quadruplex DNA and antitumor reagent were studied. Observations revealed that the complex could bind to c-myc Pu27 and Pu22 DNA strongly with constants of 1.18 × 107 (Pu27) and 4.13 × 106 M-1 (Pu22) via groove mode as determined from absorption and luminescence titrations, as well as CD spectra. Results of continuous variation analysis confirmed that the complex interacted with c-myc G-quadruplex DNA through a 1:1 binding stoichiometry. As verified by PCR-stop assay, the replication of c-myc DNA was effectively blocked by the complex with the complete inhibition at the complex concentration of 4.0 μM both for Pu27 and Pu22, suggesting that the complex could efficiently induce the formation of c-myc G-quadruplex DNA. The appearance of blue TMB solution in the visual experiment also proved that the sequences of Pu27 and Pu22 could fold into G-quadruplex under the induction of the complex. Nevertheless, the complex was found to exhibit weak stabilization ability on c-myc G-quadruplex DNA according to FRET assay, which increased the melting point of c-myc DNA only 3-3.5 °C. The experiments on Topoisomerase inhibition and cytotoxicity of the complex showed that it acted as an inhibitor of TopoI and exhibited moderate anticancer activity against MCF-7 and Huh-7 tumor cells.

Dyeing wastewater led to the water pollution and mercury contamination originated from polyvinyl chloride (PVC) production are both environmental problems derived with industrial development. In this work, the coconut activated carbon (CAC) was used to adsorb neutral red (NR), a representative dye in dyeing wastewater. Then, the spent CAC was calcined to be N-doped metal-free catalyst to realize the resource recovery. The metal-free catalyst showed a superior catalytic performance in acetylene hydrochlorination which is the important reaction for PVC production industry. The optimal 3NR/4CAC catalyst exhibited preferable catalytic activity with C2H2 conversion of 97.9% and competitive stability in the 200 h lifetime test. A series of experimental characterizations combined with ReaxFF molecular dynamics simulations as well as density functional theory (DFT) calculations have been carried out to reveal the structural and electronic properties of the N-doped CAC catalysts, the nitrogen doping process as well as the catalytic mechanism of different N species for the acetylene hydrochlorination. This work provides a novel way for the reutilization of the waste adsorbent produced from dyeing wastewater treatment to support the sustainable development of PVC industry.

To investigate the efficacy of icariin for healing skull defects in rabbit models.Thirty-six 3-month-old female New Zealand white rabbits, weighing 1.8-2.0 kg each, were randomly divided into either the control or experimental group. Skull defect models were constructed in both groups, with the experimental group receiving oral icariin afterwards. At 4, 8 and 12 postoperative weeks, the rabbits were euthanized, and X-rays and samples were taken. Tissue sections were stained using hematoxylin and eosin, followed by additional processing using histological and bone metrological methods for observing the rate and quality of the bone formation.Histologically, additional mature lamellar bone formed in the defect area in the experimental group compared with that of the control group. Bone metrological methods showed that the bone mass, trabecular bone width and number of osteoblasts in the experimental group were significantly higher than those of the control group (P0.01). The number of osteoclasts did not significantly differ between the two groups (P0.05).Icariin increased the bone mass and improved the condition of the defect area in the rabbit skulls.

Background Human immunodeficiency virus type 1 (HIV-1) infection disturbs the balance of CD4+ T cells and monocytes in the immune system. In the early stage of infection, the virus stimulates the activation and proliferation of immune cells, induces the release of cytokines, destroys CD4+ T cells, and accelerates HIV-1 replication and AIDS progression. It is essential to explore cytokine changes after HIV-1 infection and further understand the underlying mechanism of HIV infection. Methods In this study, we enrolled 38 HIV-infected subjects and 30 healthy subjects. We measured and compared CD4+ T cell counts and the serum cytokine levels in different groups. Results Our results showed significantly higher serum levels of IL-1β, IL-2, IL-4, IL-7, IL-10, IL-17, IFN-γ, and TNF-α in HIV-infected patients. Higher levels of IL-6 and IL-17 were observed in the + T cell count group, and higher levels of IL-2 were observed in the CCR5-tropic HIV strain group. Conclusion In conclusion, we found that HIV infection-induced activation of the immune system and cytokines could predict the severity of HIV disease and regulate HIV infection and replication differently depending on the type of virus strain.

This study aims to investigate the role of the long non-coding RNA (lncRNA) HOX transcript antisense RNA (HOTAIR) in the regulation of anoikis resistance of ovarian cancer cells, a prerequisite for metastasis and chemoresistance in ovarian cancer cells. Ovarian cancer SKOV3 cells were cultured in an ultra-low attachment system to establish an anoikis model. The relationship between cellular anoikis capability and HOTAIR expression level was studied by flow cytometry and RT-PCR. The ability of spheroid formation, migration, and invasion of the suspended cells was assessed following the knockdown of HOTAIR expression. The expression of EZH2, H3K27me3, representative targets of EZH2, and anoikis-related biomarkers was also detected. An increase in the duration of suspension culture time rendered the SKOV3 cells anoikis-resistant with a significantly lower apoptotic rate compared to the adherent cells. HOTAIR expression in the suspension cells increased significantly, while that in the adherent cells did not. Following small interfering RNA (siRNA)-mediated knockdown of HOTAIR expression, the abilities of anoikis resistance, migration, and invasion decreased in the suspension cells. Knockdown of HOTAIR levels also reduced the spheroid forming ability of the tumor cells in continuous suspension cultures. Moreover, EZH2 expression correlated with HOTAIR expression, thus regulating the expression of miR-193a and DOK2 via introducing H3K27me3. Western blot analysis of anoikis-related markers showed that N-cadherin, ZEB1, and TWIST1 were downregulated following inhibition of HOTAIR, while E-cadherin and ErbB3 were upregulated. In conclusion, HOTAIR enhances the anoikis resistance and spheroid forming ability of ovarian cancer cells by recruiting EZH2 and influencing H3K27 methylation that may contribute to migration, invasion, and chemoresistance of ovarian cancer cells.

Background Over the past decade, increasing attention has been paid on post-stroke suicide (PSS), which is one of complications of stroke. The rates of stroke and suicide are relatively high, especially in Asian populations. Thus, a deeper understanding of the prevalence and epidemiological impact of suicide after stroke is urgently needed. Clinical diagnosis and prevention of PSS are at the incipient stage, but the risk factors responsible for the occurrence of PSS in different regions and stages of the disease remain largely unknown. The present meta-analysis aimed to determine the incidence of PSS at different stages and time courses, and to identify the underlying risk factors for PSS. Methods We systematically searched the Cochrane library, Embase, PubMed,CNKI and Web of Science databases from their inception until April 2019. The research articles reporting on the risk factor for PSS were screened and included in the meta-analysis. The data from the included studies were extracted according to the predefined criteria. Results A total of 12 studies (n = 2693036) were included for meta-analyses. Of these studies, 7 reported suicide prevalence were meta-analyzed. The pooled estimate of suicidal ideation rates after stroke was 12%, which could be influenced by multiple risk factors, including men, smoking, depression, sleep disorders, previous stroke and low household income. Studies conducted in Asia demonstrated higher suicide prevalence (approximately 15%) compared to other regions. Smoking, alcohol consumption, low family income, depression, heart disease and sleep disorders were the important risk factors for the pathogenesis of suicide after stroke. When compared to PSS more than one year, the incidence of suicide within one year after stroke was more likely to be statistically significant. It was found that 4 out of every 1,000 stroke survivors tended to commit suicide. The results of this meta-analysis showed that depression (OR = 2.32; p