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Qi Jin1 *, Jie Li2 *, Guang-Yao Chen1 *, Zi-Yu Wu,3 Xiao-Yu Liu,2 Yi Liu,4 Lin Chen,5 Xin-Yi Wu,5 Yan Liu,2 Xin Zhao,2 Yue-Han Song2 1Graduate School, Beijing University of Chinese Medicine, Beijing, 100029, People’s Republic of China; 2School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, 100029, People’s Republic of China; 3Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, 100007, People’s Republic of China; 4Humanities School, Beijing University of Chinese Medicine, Beijing, 100029, People’s Republic of China; 5Qihuang School, Beijing University of Chinese Medicine, Beijing, 100029, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xin Zhao; Yue-Han Song, Email zhaoyyyx@sohu.com; songyuehan1981@126.comObjective: The mechanism of Wendan Decoction (WDD) against Generalized Anxiety Disorder (GAD) was predicted by network pharmacology and validated by in vivo and in vitro experiments.Methods: The targets of WDD for the treatment of GAD were obtained by a search of online databases. Further, PPI network and KEGG enrichment were used to identify the key targets and pathways. Ultimately, these key targets and pathways were validated by in vivo experiments on GAD mice modeled by repeated restraint stress (RRS) and in vitro experiments on inflammatory factor stimulated BV-2 cells.Results: Through searching the databases, the 137 ingredients of WDD that correspond to 938 targets and 4794 targets related to GAD were identified. Among them, 569 overlapping targets were considered as the therapeutic targets of WDD for GAD. PPI analysis showed that the inflammation-related proteins IL-6, TNF, SRC and AKT1 were the key targets, and KEGG enrichment suggested that PI3K/AKT and MAPK signaling pathways were key pathways of WDD in the treatment of GAD. In vivo experiments, RRS mice exhibited abnormality in behavioristics in open field test (OFT) and elevated plus maze (EPM) and increases in serum corticosterone and the percentage of lymphocytes positive for IL-6 in peripheral blood. These abnormal changes can be reversed by WDD and the positive control drug paroxetine. In vitro experiments, WDD can inhibit IL-6 induced activation of PI3K/AKT and MAPK signaling pathways in BV2 cells, and suppress the ensuing release of inflammatory factors TNF-α, IL-1β and PGE2, and showed a dose-dependent effect.Conclusion: WDD is able to resist GAD by relieving inflammatory response in peripheral and central system.Keywords: Wendan Decoction, Generalized Anxiety Disorder, network pharmacology, Interleukin-6, PI3K/AKT signaling pathway, MAPK signaling pathway

Bulbus eleutherinis is a classical traditional Dai medicine, and has been widely used in clinical treatment of coronary heart disease (CHD) in Yunnan, China. Naphthoquinone, as the main active compound in Bulbus eleutherinis in treating CHD, mainly contain isoeleutherin, eleutherin, and eleutherol. This study aimed to investigate the in vivo parameters of isoeleutherin, eleutherin, and eleutherol. In this work, male Sprague Dawley (SD) rats were treated with the three compounds by oral administration, and then blood and tissue samples were collected. A novel UPLC-MS/MS (ultra-performance liquid chromatography-tandem mass spectrometry) method has been developed to determine the absolute oral bioavailability, and the tissue distribution profile of the compounds. Acetonitrile and 0.1% (v/v) solution of formic acid were selected as the mobile phase of the chromatogram. C18 column was employed. Betamethasone was used as an internal standard in the method. The detection was performed with a multireaction monitor of scan type in positive ion mode by MS/MS. Our data showed linearity of the method with r over 0.9983. Lower limits of quantification of isoeleutherin, eleutherin, and eleutherol were 1.00, 3.84, and 0.498 ng/mL, respectively. The overall precision of the compounds was less than 12.68%, recoveries ranged from 85.44 to 103.83%, and the accuracy of the compounds in plasma was between 91.56 and 110.75%. The stability assay showed that they were stable (87.83–114.62%) under different conditions in plasma. For oral administration, the half-lives of isoeleutherin, eleutherin, and eleutherol was 6.11, 7.30, and 3.07 hours, respectively. The absolute oral bioavailabilities were 5.38, 4.64, and 2.47%, respectively. Moreover, the three components had the highest distribution in small intestine. In conclusion, the established method was successfully applied to the determination of the in vivo parameters of the three components in SD rats. This work provides a reference for the development of new drugs of Bulbus eleutherinis in the future.

Cymbidium, which includes approximately 80 species, is one of the most ornamental and cultivated orchid genera. However, a lack of markers and sparse sampling have posed great challenges to resolving the phylogenetic relationships within the genus. In the present study, we reconstructed the phylogenetic relationships by utilizing one nuclear DNA (nrITS) and seven plastid genes (rbcL, trnS, trnG, matK, trnL, psbA, and atpI) from 70 species (varieties) in Cymbidium. We also examined the occurrence of phylogenetic conflict between nuclear (nrITS) and plastid loci and investigated how phylogenetic conflict bears on taxonomic classification within the genus. We found that phylogenetic conflict and low support values may be explained by hybridization and a lack of informative characteristics. Our results do not support previous classification of the subgenera and sections within Cymbidium. Discordance between gene trees and network analysis indicate that reticulate evolution occurred in the genus Cymbidium. Overall, our study indicates that Cymbidium has undergone a complex evolution., Highlights • A comprehensive understanding of Cymbidium phylogeny. • Reticulate evolution occurred in the genus Cymbidium. • Cymbidium has a complex evolution.

Dendrobium catenatum (Dendrobium officinale) is a valuable genuine herb. The source of this species is difficult to be identified by traditional methods including morphology, spectroscopy, and chromatography. We used the restriction site-associated DNA sequencing (RAD-seq) approach to perform the high-throughput sequencing of 24 D. catenatum provenances. In this study, 371.18 Gb clean data were obtained, and 655,057 high-quality SNPs were selected after their filtration. We used phylogenetic tree, genetic structure, and principal component analyses to examine the genetic diversities and genetic relationships of the 109 accessions. We found that D. catenatum could be divided into two groups, and each group was closely related to the distribution of the sampling sites. At the population level, the average nucleotide diversity (π) of the D. catenatum population mutation parameters was 0.1584 and the expected heterozygosity (HE) was 0.1575. The GXLPTP07 accessions showed the highest genetic diversity in terms of the private allele number, observed heterozygosity, and nucleotide diversity. The Mantel test showed a significant positive correlation between the genetic and geographic distances among the overall distribution. A genetic information database of D. catenatum was established, which confirmed that RAD-seq technology has the potential to be applied in the identification of medicinal Dendrobium of different origins.

AIM: To assess the patient-reported outcome of phacoemulsification with posterior chamber single focus foldable intraocular lens (SIOL) using the Chinese version of Catquest-8SF questionnaire (Catquest-8SF-CN), evaluate the applicability of Catquest-8SF scale in northern Chinese and its sensitivity to cataract surgery. METHODS: Prospective clinical case follow-up study. The patients were recruited from the Central Hospital of Tai’an and completed the Catquest-8SF questionnaire by face-to-face interviews before and 3mo after surgery at the hospital. RESULTS: A total of 120 cataract patients with median age 67.6y and 52.5% female completed the Catquest-8SF. The Cronbach’s α coefficient of the total scale, daily-activity sub-table and comprehensive-evaluation sub-table were 0.861, 0.853 and 0.748 respectively. There was a high level of consistency between two investigators. The cumulative contribution rate was 66.64% by using the principal component analysis and the maximum variance orthogonal rotation method. The preoperative score of the Catquest-8SF-CN had a significant negative correlation with preoperative binocular mean weighted visual acuity and age (P

Laboratory scale recycling of marine plastic litter consisting of polyethylene terephthalate (PET) bottle sorting, pyrolysis and chemical vapor deposition (CVD) was conducted to identify the technical and environmental implications of the technology when dealing with real waste streams. Collected seashore and underwater plastics (SP and UP, respectively) contained large quantities of PET bottles (33.2 wt% and 61.4 wt%, respectively), suggesting PET separation was necessary prior to pyrolysis. After PET sorting, marine litter was converted into pyrolysis oil and multi-walled carbon nanotubes (MWCNTs). Water-based washing of litter prior to pyrolysis did not significantly change the composition of pyrolysis products and could be avoided, eliminating freshwater consumption. However, distinct differences in oil and MWCNT properties were ascribed to the variations in feedstock composition. Maintaining consistent product quality would be one of challenges for thermochemical treatment of marine litter. As for the environmental implications, life cycle assessment (LCA) demonstrated positive benefits, including improved climate change and fossil depletion potentials. The highest positive environmental impacts were associated with MWCNT production followed by pyrolysis oil and PET recovery. The benefits of proposed approach combining PET sorting, pyrolysis and CVD allowed to close the waste loop by converting most of the marine litter into valuable products. Economic Development Board (EDB) Nanyang Technological University The authors would also like to acknowledge the Nanyang Environment and Water Research Institute, Nanyang Technological University (Singapore) and Economic Development Board (Singapore) for financial support of this research.

Objectives. Although a recent study reported that stimulator of interferon genes (STING) in macrophages has an important regulatory effect on liver ischemia-reperfusion injury (IRI), the underlying mechanism of STING-dependent innate immune activation in liver macrophages (Kupffer cells, KCs) remains unclear. Here, we investigated the effect of STING on liver macrophage pyroptosis and the associated regulatory mechanism of liver IRI. Methods. Clodronate liposomes were used to block liver macrophages. AAV-STING-RNAi-F4/80-EGFP, an adenoassociated virus (AAV), was transfected into the portal vein of mice in vivo, and the liver IRI model was established 14 days later. In vitro, liver macrophages were treated with STING-specific siRNA, and a hypoxia-reoxygenation (H/R) model was established. The level of STING was detected via Western blotting (WB), RT-PCR, and immunostaining. Liver tissue and blood samples were collected. Pathological changes in liver tissue were detected by hematoxylin and eosin (H&E) staining. Macrophage pyroptosis was detected by WB, confocal laser scanning microscopy (CLSM), transmission electron microscopy (TEM), and enzyme-linked immunosorbent assay (ELISA). The calcium concentration was measured by immunofluorescence and analyzed with a fluorescence microplate reader. Results. The expression of STING increased with liver IRI but decreased significantly after the clodronate liposome blockade of liver macrophages. After knockdown of STING, the activation of caspase 1-GSDMD in macrophages and liver IRI was alleviated. More interestingly, hypoxia/reoxygenation (H/R) increased the calcium concentration in liver macrophages, but the calcium concentration was decreased after STING knockdown. Furthermore, after the inhibition of calcium in H/R-induced liver macrophages by BAPTA-AM, pyroptosis was significantly reduced, but the expression of STING was not significantlydecreased. Conclusions. Knockdown of STING reduces calcium-dependent macrophage caspase 1-GSDMD-mediated liver IRI, representing a potential therapeutic approach in the clinic.

Thyroid hormone receptor interactor 13 (TRIP13) plays a crucial role in poor prognosis and chemotherapy resistance of cancer patients. This present study is aimed at investigating the role of high expression of TRIP13 inducing nedaplatin (NDP) resistance in esophageal squamous cell carcinoma (ESCC) cells. High expression of TRIP13 promoted the proliferation and migration of ESCC cells performed by MTS assay, colony formation assay, wound healing assay, and transwell assay. High TRIP13 expression induced NDP resistance to ESCC based on the cell proliferation promoting/inhibition rate and cell migration promoting/inhibition rate analysis, flow cytometry assay of apoptotic subpopulations with a combination of Annexin V-FITC and propidium iodide, and Western blot analysis downregulating cleaved PARP, γH2A.X, cleaved caspase-3, and Bax and upregulating Bcl-2 expression. This study indicated that high expression of TRIP13 promoted proliferation and migration of ESCC cells and induced NDP resistance via enhancing repair of DNA damage and inhibiting apoptosis. This will provide a preliminary reference for the clinical use of NDP in ESCC treatment.

Tetraphosphine ligand dpppda reacts with AgNO3 to generate two 1D coordination polymers containing cubic and chair-like [Ag4(NO3)4] clusters. ESR spectroscopy supported by DFT calculations indicated the presence of dpppda˙+, a Wurster's blue - like radical cation formed by intramolecular ligand-to-metal charge transfer (LMCT). This LMCT results in higher AgI → Ag0 potentials and good photocurrent responses.

To study the effects of total ginsenosides (TG) extract from Panax ginseng on neural stem cell (NSC) proliferation and differentiation and their underlying mechanisms.The migration of NSCs after treatment with various concentrations of TG extract (50, 100, or 200 µ g/mL) were monitored. The proliferation of NSCs was examined by a combination of cell counting kit-8 and neurosphere assays. NSC differentiation mediated by TG extract was evaluated by Western blotting and immunofluorescence staining to monitor the expression of nestin and microtubule associated protein 2 (MAP2). The GSK-3β/β-catenin pathway in TG-treated NSCs was examined by Western blot assay. The NSCs with constitutively active GSK-3β mutant were made by adenovirus-mediated gene transfection, then the proliferation and differentiation of NSCs mediated by TG were further verified.TG treatment significantly enhanced NSC migration (P0.01 or P0.05) and increased the proliferation of NSCs (P0.01 or P0.05). TG mediation also significantly upregulated MAP2 expression but downregulated nestin expression (P0.01 or P0.05). TG extract also significantly induced GSK-3β phosphorylation at Ser9, leading to GSK-3β inactivation and, consequently, the activation of the GSK-3β/β-catenin pathway (P0.01 or P0.05). In addition, constitutive activation of GSK-3β in NSCs by the transfection of GSK-3β S9A mutant was found to significantly suppress TG-mediated NSC proliferation and differentiation (P0.01 or P0.05).TG promoted NSC proliferation and neuronal differentiation by inactivating GSK-3β.

WebComparing the whole genome sequence of Apostasia shenzhenica with transcriptome and genome data from five orchid subfamilies permits the reconstruction of an ancestral gene toolkit, providing insight into orchid origins, evolution and diversification. Supplementary information The online version of this article (doi:10.1038/nature23897) contains supplementary material, which is available to authorized users., Orchid origins Around 10 per cent of flowering plant species are orchids, with a broad diversity in both morphology and lifestyle. Apostasia is one of the earliest-diverging genera of Orchidaceae. To study the evolution and diversity of Orchidaceae, Zhong-Jian Liu, Yves Van de Peer and colleagues sequenced the genome of Apostasia shenzhenica, a self-pollinating species found in southeast China. The authors also report improved genomes for two species of Epidendroideae, Phalaenopsis equestris and Dendrobium catenatum, as well as transcriptome analysis of representatives of subfamilies of Orchidaceae. Their analyses provide insights into orchid origins, genome evolution, adaptation and diversification. Supplementary information The online version of this article (doi:10.1038/nature23897) contains supplementary material, which is available to authorized users., Constituting approximately 10% of flowering plant species, orchids (Orchidaceae) display unique flower morphologies, possess an extraordinary diversity in lifestyle, and have successfully colonized almost every habitat on Earth1,2,3. Here we report the draft genome sequence of Apostasia shenzhenica4, a representative of one of two genera that form a sister lineage to the rest of the Orchidaceae, providing a reference for inferring the genome content and structure of the most recent common ancestor of all extant orchids and improving our understanding of their origins and evolution. In addition, we present transcriptome data for representatives of Vanilloideae, Cypripedioideae and Orchidoideae, and novel third-generation genome data for two species of Epidendroideae, covering all five orchid subfamilies. A. shenzhenica shows clear evidence of a whole-genome duplication, which is shared by all orchids and occurred shortly before their divergence. Comparisons between A. shenzhenica and other orchids and angiosperms also permitted the reconstruction of an ancestral orchid gene toolkit. We identify new gene families, gene family expansions and contractions, and changes within MADS-box gene classes, which control a diverse suite of developmental processes, during orchid evolution. This study sheds new light on the genetic mechanisms underpinning key orchid innovations, including the development of the labellum and gynostemium, pollinia, and seeds without endosperm, as well as the evolution of epiphytism; reveals relationships between the Orchidaceae subfamilies; and helps clarify the evolutionary history of orchids within the angiosperms. Supplementary information The online version of this article (doi:10.1038/nature23897) contains supplementary material, which is available to authorized users.

We analyze the high-spin structure of the even–even 72–80Kr isotopes using the Projected Shell Model (PSM). With the help of the Pfaffian formulas, we have vigorously extended the quasi-particle (qp) basis of the PSM code and applied in this mass region for the first time. We consider a sufficiently large multi-qp configuration space in order to describe high-spin rotational behavior. The results show that the calculation can reproduce most of the known rotational bands with positive- or negative-parity. Moreover, some side bands appearing in the near-yrast region are predicted. The main structure for each band is discussed in terms of multi-qp configurations. The variations in moment of inertia with spin are explained in terms of successive band crossings among the 2-qp, 4-qp, 6-qp, and 8-qp states. The B ( E 2 ) transition probabilities in these bands are also calculated. To further understand the high-spin behavior of these neutron-deficient nuclei and to confirm predictions of the present work, good high-spin data, especially for B ( E 2 ) transitions, are called for.

Constituting approximately 10% of flowering plant species, orchids (Orchidaceae) display unique flower morphologies, possess an extraordinary diversity in lifestyle, and have successfully colonized almost every habitat on Earth. Here we report the draft genome sequence of Apostasia shenzhenica, a representative of one of two genera that form a sister lineage to the rest of the Orchidaceae, providing a reference for inferring the genome content and structure of the most recent common ancestor of all extant orchids and improving our understanding of their origins and evolution. In addition, we present transcriptome data for representatives of Vanilloideae, Cypripedioideae and Orchidoideae, and novel third-generation genome data for two species of Epidendroideae, covering all five orchid subfamilies. A. shenzhenica shows clear evidence of a whole-genome duplication, which is shared by all orchids and occurred shortly before their divergence. Comparisons between A. shenzhenica and other orchids and angiosperms also permitted the reconstruction of an ancestral orchid gene toolkit. We identify new gene families, gene family expansions and contractions, and changes within MADS-box gene classes, which control a diverse suite of developmental processes, during orchid evolution. This study sheds new light on the genetic mechanisms underpinning key orchid innovations, including the development of the labellum and gynostemium, pollinia, and seeds without endosperm, as well as the evolution of epiphytism; reveals relationships between the Orchidaceae subfamilies; and helps clarify the evolutionary history of orchids within the angiosperms.

Dendrobium harveyanum is an endangered species of Orchidaceae. Here we report the complete chloroplast (cp) genome sequence and the cp genome features of D. harveyanum. The complete cp genome sequence of D. harveyanum is 157,292 bp in length and presented a typical quadripartite structure including one large single-copy region (LSC, 86,583 bp), one small single-copy region (SSC, 19,449 bp), and two inverted repeat regions (IRs, 25,630 bp each). The cp genome encoded 138 genes, of which 120 were unique genes. The phylogenetic relationships show that D. harveyanum is closely related to other species in Dendrobium.

Cymbidium mastersii Griff. & Lindl. is an endangered orchid. In this study, we report the complete chloroplast (cp) genome sequence and the cp genome features of C. mastersii. The cp genome sequence of C. mastersii was 155,362 bp in length. It included one large single-copy region (LSC, 84,465 bp), one small single-copy region (SSC, 20,647 bp), and two inverted repeat regions (IRs, 25,125 bp). The cp genome encoded 130 genes, of which 107 were unique genes (80 protein-coding genes, 23 tRNAs, and 4 rRNAs). The maximum-likelihood phylogenetic analysis showed that C. mastersii was a sister of C. erythraeum and C. nanulum.

Cymbidium eburneum Lindl. is an endangered species of Orchidaceae and distributed in Guangxi, Yunnan and Hainan of China, and India, Myanmar, Nepal, Vietnam. Here, we report the complete chloroplast (cp) genome sequence and the cp genome features of C. eburneum. The complete chloroplast (cp) genome sequence of C. eburneum is 156,520 bp in length and presented a typical quadripartite structure including one large single-copy region (LSC, 85,518 bp), one small single-copy region (SSC, 20,014 bp), and two inverted repeat regions (IRs, 25,471 bp). The cp genome encoded 135 genes, of which 106 were unique genes (78 protein-coding genes, 24 tRNAs, and 4 rRNAs). The phylogenetic relationships show that C. eburneum is closely related to other species in the genus Cymbidium, and is sister with C. tracyanum.

Reaction of AgNO 3 with one tetraphosphine ligand, 1,4- N , N , N ′, N ′-tetra(diphenylphosphanylmethyl) benzene diamine (dpppda), gave rise to a Ag(I)/tetraphosphine coordination polymer [Ag 4 (NO 3 ) 4 (dpppda)] n ( 1 ). Compound 1 has a one-dimensional (1D) chain structure in which the chair-like [Ag 4 (NO 3 ) 4 ] cores are linked by the dpppda ligands using a Z-shaped μ - η 2 : η 2 side-by-side mode. Compound 1 exhibited good catalytic activity towards the photodegradation of nitrobenzene (NB), paranitrophenol (PNP) and 2,4-dinitrophenol (2,4-DNP) in aqueous solution under UV light irradiation. The kinetics and the mechanism of such catalytic photodegradation reactions were also investigated. The present work provided some new insight into the design and preparation of new coordination polymers as catalyst for high-performance photodegradation of toxic and persistent organic species existed in industrial wastewaters.

Background: The interplay between collective and single-particle degrees of freedom is an important structure aspect to study. The nuclei in theA ≈ 180 mass region are often denoted as good examples to study such problems because these nuclei are known to exhibit many rotational bands based on multi-quasiparticle K isomers. Purpose: A large set of high-quality experimental data on high-K isomeric states in the A ≈ 180 mass region has accumulated. A systematic description of them is a theoretical challenge as it requires a method going beyond the usual mean field with multi-quasiparticle configurations built in the shell-model basis. The K-isomer data provide an ideal testing ground for theoretical models. Method: The recently extended projected shell model (PSM) by the Pfaffian method is employed with a sufficiently large configuration space including up to 10 quasiparticles. The restoration of rotational symmetry which is broken in the deformed mean field is obtained by means of angular-momentum projection. With axial symmetry in the basis deformation, each multi-quasiparticle state, classified by a K quantum number, represents the major component of a rotational K band. Shell-model diagonalization in such a projected basis defines the K mixing, which is the key ingredient of the present method. Results: Quasiparticle structure and rotational properties of high-K isomers in even-even neutron-rich 174−186W isotopes are described. The rotational evolution of the yrast and near-yrast bands is discussed with successive band crossings. Multi-quasiparticle K isomers and associated rotational bands in each W isotope are studied with detailed quasiparticle configurations given. Electromagnetic transition properties are also studied and the calculated B(E2), B(M1), and g-factors are compared with experiment if data exist. Conclusions: Many nuclei of the A ≈ 180 mass region exhibit properties of an axially symmetric shape and K is approximately a good quantum number. For such nuclei, the extended PSM assuming an axially symmetric basis but including K mixing through diagonalization of the two-body Hamiltonian is an appropriate method to study multi-quasiparticle K isomers and K violations in these states. For special examples where one finds highly K-forbidden transitions the present model needs to be further improved.

Reactions of the tetraphosphine ligand N,N,N',N'-tetra(diphenylphosphanylmethyl)ethylene diamine) (dppeda) or 1,4-N,N,N',N'-tetra(diphenylphosphanylmethyl)benzene diamine (dpppda) with NiCl2, ancillary ligand (C2H5)2NCS2Na (Na(dtc)) and NH4PF6 afforded two binuclear complexes [(dtc)Ni(dppeda)Ni(dtc)](PF6)2 (1) and [(dtc)Ni(dpppda)Ni(dtc)](PF6)2 (2). Compounds 1 and 2 were characterized by elemental analyses, IR spectra, (1)H, (13)C{(1)H} NMR and (31)P{(1)H} NMR, electrospray ionization (ESI) mass spectra, thermogravimetric (TGA) experiments and single crystal X-ray diffraction. The Ni(ii) atoms in both compounds form a square planar coordination geometry while the ligands hold a similar end-to-end coordination mode. Both products can catalyze the ring-opening polymerization (ROP) of ε-caprolactone (ε-CL) while 1 exhibited a higher catalytic activity than 2. Influencing aspects such as solvent, temperature, the ratio of ε-CL to catalyst, the presence of benzyl alcohol, concentration of ε-CL and reaction intervals were investigated.

Delayed healing of corneal epithelial wounds is a serious complication in diabetes. Advanced glycation end products (AGEs) are intimately associated with the diabetic complications and are deleterious to the wound healing process. However, the effect of AGEs on corneal epithelial wound healing has not yet been evaluated. In the present study, we investigated the effect of AGE-modified bovine serum albumin (BSA) on corneal epithelial wound healing and its underlying mechanisms. Our data showed that AGE-BSA significantly increased the generation of intracellular ROS in telomerase-immortalized human corneal epithelial cells. However, the generation of intracellular ROS was completely inhibited by antioxidant N-acetylcysteine (NAC), anti-receptor of AGEs (RAGE) antibodies, or the inhibitor of NADPH oxidase. Moreover, AGE-BSA increased NADPH oxidase activity and protein expression of NADPH oxidase subunits, p22phox and Nox4, but anti-RAGE antibodies eliminated these effects. Furthermore, prevention of intracellular ROS generation using NAC or anti-RAGE antibodies rescued AGE-BSA-delayed epithelial wound healing in porcine corneal organ culture. In conclusion, our results demonstrated that AGE-BSA impaired corneal epithelial wound healing ex vivo. AGE-BSA increased intracellular ROS generation through NADPH oxidase activation, which accounted for the delayed corneal epithelial wound healing. These results may provide better insights for understanding the mechanism of delayed healing of corneal epithelial wounds in diabetes.

Reactions of N,N-bis(diphenylphosphanylmethyl)-2-aminopyridine (bdppmapy) with [Ag(MeCN)4]ClO4 or AgX (X = Cl, Br, I, SCN, CN) afforded a family of oligomeric and polymeric complexes: [Ag2(MeOH)(bdppmapy)2](ClO4)2 (1), [AgCl(bdppmapy)] (2), [AgBr(bdppmapy)] (3), [AgI(bdppmapy)] (4), [AgSCN(bdppmapy)] (5), [{(η2-bdppmapy)Ag(μ-CN)AgCN}2(μ-bdppmapy)] (6), [Ag4(μ-CN)4(μ-bdppmapy)] (7), and [Ag2(μ-CN)(μ-bdppmapy)2][Ag5(μ-CN)6] (8). Compounds 1–8 were characterized by elemental analyses, IR spectra, 1H and 31P{1H} NMR, electrospray ionization (ESI) mass spectra, powder X-ray diffraction (XRD), and single-crystal X-ray diffraction. Compounds 1–3 and 5 hold a one-dimensional (1D) chain in which [Ag(MeOH)]+ or [AgX] motifs are linked by bdppmapy bridges. Compound 6 has a tetrameric framework in which two linear [(η2-bdppmapy)Ag(μ-CN)AgCN] fragments are connected by a μ-bdppmapy ligand. Compound 7 contains a 1D staircase chain in which two zigzag [Ag(μ-CN)]n chains are linked by pairs of μ-bdppmapy bridges. Compoun...

Isorhamnetin, a 3‑O‑methylated metabolite of quercetin, exhibits antioxidant effects. However, to the best of our knowledge, no study to date has focused on the effects of isorhamnetin on retinal pigment epithelium (RPE) cells, and its underlying molecular mechanisms. Therefore, the present study aimed to examine the potential protective effect of isorhamnetin against oxidative stress in human RPE cells. The results demonstrated that pretreatment of RPE cells with isorhamnetin significantly protected cell viability against oxidative stress. In addition, isorhamnetin pretreatment inhibited hydrogen peroxide (H2O2)‑induced reactive oxygen species (ROS) production and caspase‑3 activation in RPE cells. Furthermore, isorhamnetin pretreatment significantly increased the phosphorylation of phosphoinositide 3‑kinase (PI3K) and AKT serine/threonine kinase 1 (Akt) in RPE cells exposed to H2O2, compared with cells treated with H2O2 alone. Taken together, the present results demonstrated that isorhamnetin protected human RPE cells from oxidative stress‑induced cell death, and this effect was associated with activation of the PI3K/Akt signaling pathway. Thus, isorhamnetin may be considered as a potential antioxidant useful for the prevention of age‑related macular degeneration.

To study the inhibition of nuclear factor kappa-B p65 (NF-κB p65) antisense oligodeoxynucleotide (ASODN) on transdifferentiation of normal human lens epithelial cells induced by transforming growth factor-β2 (TGF-β2) in vitro.NF-κB p65 ASODN and NF-κB p65 missense oligodeoxynucleotide (MSODN) were designed and synthesized. Human lens epithelial cell line (HLE B-3) cells were prepared for study and divided into 7 groups. Control group was HLE B-3 cells cultured in vitro in dulbecco's modified eagle medium (DMEM). T1, T2, and T3 group were HLE B-3 cells cultured in vitro in DMEM with 10 ng/mL TGF-β2 for 6h, 12h, 24h respectively. A+T group was HLE B-3 cells cultured with 10 ng/mL TGF-β2 for 24h after transfected by NF-κB p65 ASODN for 24h. M+T group was HLE B-3 cells cultured with 10 ng/mL TGF-β2 for 24h after transfected by NF-κB p65 MSODN for 24h. The negative control group was HLE B-3 cells cultured with 10 ng/mL TGF-β2 for 24h after cultured with transfer agent (HiPerFect) for 24h. Cell morphology was observed at different time points using an inverted microscope. The expression of NF-κB p65 mRNA was detected with reverse transcription-polymerase chain reaction (RT-PCR), and the expression of α-smooth muscle actin (α-SMA) protein was assayed with ELISA.With the TGF-β2 stimulation prolongation, the expression of NF-κB p65 mRNA and α-SMA protein increased in T1, T2, T3 groups compared with the control group, and the difference was statistically significant (P0.05). NF-κB p65 ASODN lowered the expression of NF-κB p65 mRNA and α-SMA protein induced by TGF-β2. NF-κB p65 MSODN and HiPerFect did not lower the expression of NF-κB p65 mRNA and α-SMA protein induced by TGF-β2. The difference between control group and A+T group was not statistically significant (P0.05), but the difference among A+T group and other groups was statistically significant (P0.05).NF-κB p65 ASODN could lower the expression of NF-κB p65 mRNA and α-SMA protein induced by TGF-β2, and antagonized TGF-β2-induced transdifferentiation of HLE B-3 in vitro. NF-κB p65 ASODN could be used as a new biological therapeutic target of posterior capsular opacification.

A new orchid species, Cymbidium densiflorum , from Yunnan, China, is described and illustrated here based on morphological and molecular analyses. A detailed comparison between the newly discovered orchid and other members of Cymbidium was conducted. The new plant belongs to C . section Cymbidium and has a pendent inflorescence, thick-coriaceous flowers with two pollinia and rigid leaves, and it differs greatly from all other known species of this section by being a large plant bearing leaves with an acute, oblique apex and producing a long inflorescence with more than 30 purplish brown large flowers, in which the lateral sepals are oblique and narrowly elliptic with a tooth at the apex and narrowly ovate-elliptic petals . A molecular study based on the nuclear ribosomal ITS and plastid matK , rbcL , trnG , trnL and atpI DNA sequence data indicated that C. densiflorum is sister to C. paucifolium .

Dendrobium libingtaoi is described from southeastern Yunnan, China, based on morphological and DNA evidence. In the morphological analysis, D. libingtaoi is similar to D. draconis but differs in having ovate-elliptical petals, a lip with a light red blotch in the middle, oblong-lanceolate lateral sepals, and a nearly round midlobe. Nuclear and plastid DNA analyses indicate that D. libingtaoi is a member of D. section Formosae.

Slim pump for cooling devices play an important role for cooling the electronics apparatus. This paper develops an axial-flux internal rotor type motor for the low-profile vortex pump. The integrated motor-pump structure is composed of an internal permanent magnet (PM) disc rotor and double-sided external coreless stators with windings piled up on the printed circuit board (PCB). The impeller is disposed on the periphery of the magnet and a fluid dynamic bearing (FDB) is set in the centre of the magnet to rotate relative to the centre shaft. A prototype of the axial-flux motor pump and drive circuit has been made and tested. Both simulated and experimental results show that the proposed motor would be an acceptable solution for the slim notebook (NB) applications.

BACKGROUND Cornea epithelial cells play early and crucial roles in the initiation of ocular surface responses to pathogens. Participation of toll-like receptor (TLR) 2 and TLR4, which are major forms of fungi receptors, may be involved in Aspergillus fumigatus induced immune responses. The objective of the present study was to examine whether inactive Aspergillus fumigatus conidia induce NF-kappaB activation and production of proinflammatory cytokines, and whether the expression of TLR2 and TLR4 were amplified by conidia in cultured immortalized human corneal epithelial cells (THCEs). This may contribute to our knowledge of the mechanism by which the host cornea can successfully defend against invasive fungi. METHODS Aspergillus fumigatus conidia were used to challenge THCE cells. THCE cells were harvested after 0.5, 1, 2 or 4 hours incubation. Real-time quantitative PCR was performed to determine the expression of TLR2, TLR4, TNF-alpha and IL-8. Western blotting was performed to determine the expression of NF-kappaB. Enzyme-linked immunosorbent assay (ELISA) was performed to determine the expression of TNF-alpha and IL-8. And the release of TNF-alpha and IL-8 in the cell supernatant were also assessed by ELISA with or without pretreatment with TLR2 and TLR4 neutralizing antibodies. RESULTS Aspergillus fumigatus conidia elicited the expression of TLR2, TLR4, TNF-alpha and IL-8 mRNA in THCEs. Exposure of THCE cells to Aspergillus fumigatus conidia resulted in NF-kappaB activation, which increased at 30 minutes (increased from 11.35+/-2.74 in the controls to 19.12+/-3.48, P

A hybrid numerical method involving the Laplace transform technique and finite-difference method in conjunction with the least-squares method and actual experimental temperature data inside the test material is proposed to estimate the unknown surface conditions of inverse heat conduction problems with the temperature-dependent thermal conductivity and heat capacity. The nonlinear terms in the differential equations are linearized using the Taylor series approximation. In this study, the functional form of the surface conditions is unknown a priori and is assumed to be a function of time before performing the inverse calculation. In addition, the whole time domain is divided into several analysis subtime intervals and then the unknown estimates on each subtime interval can be predicted. In order to show the accuracy and validity of the present inverse scheme, a comparison among the present estimates, direct solution, and actual experimental temperature data is made. The effects of the measurement errors, ...