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Nitrogen (N) is the main nutrient of plants, and low nitrogen usually affects plant growth and crop yield. The traditional Chinese herbal medicine Dendrobium officinale Kimura et. Migo is a typical low nitrogen-tolerant plant, and its mechanism in response to low nitrogen stress has not previously been reported. In this study, physiological measurements and RNA-Seq analysis were used to analyse the physiological changes and molecular responses of D. officinale under different nitrogen concentrations. The results showed that under low nitrogen levels, the growth, photosynthesis and superoxide dismutase activity were found to be significantly inhibited, while the activities of peroxidase and catalase, the content of polysaccharides and flavonoids significantly increased. Differentially expressed genes (DEGs) analysis showed that nitrogen and carbon metabolisms, transcriptional regulation, antioxidative stress, secondary metabolite synthesis and signal transduction all made a big difference in low nitrogen stress. Therefore, copious polysaccharide accumulation, efficient assimilation and recycling of nitrogen, as well as rich antioxidant components play critical roles. This study is helpful for understanding the response mechanism of D. officinale to low nitrogen levels, which might provide good guidance for practical production of high quality D. officinale.

IiSVP of Isatis indigotica was cloned and its expression pattern was analyzed. Ectopic expression of IiSVP in Arabidopsis could delay the flowering time and reduce the size of the floral organs. SVP (SHORT VEGETATIVE PHASE) can negatively regulate the flowering time of Arabidopsis. In the present work, the cDNA of IiSVP, an orthologous gene of AtSVP in I. indigotica, was cloned. IiSVP was highly expressed in rosette leaves, inflorescences and petals, but weakly expressed in sepals, pistils and young silicles. The results of subcellular localization showed that IiSVP was localized in nucleus. Bioinformatics analysis indicated that this protein was a MADS-box transcription factor. Constitutive expression of IiSVP in Arabidopsis thaliana resulted in decrease of the number of petals and stamens, and curly sepals were formed. In IiSVP transgenic Arabidopsis plants, obvious phenotypic variations in flowers could be observed, especially the size of the floral organs. In comparison with the wild-type plants, the size of petals, stamens and pistil in IiSVP transgenic Arabidopsis plants was decreased significantly. In some transgenic plants, the petals were wrapped by the sepals. Yeast two-hybrid experiments showed that IiSVP could form higher-order complexes with other MADS proteins, including IiSEP1, IiSEP3, IiAP1 and IiSEP4, but could not interact with IiSEP2. In this work, it was proved that the flowering process and the floral development in Arabidopsis could be affected by IiSVP from I. indigotica Fortune.

The common skeletons and formulations of hydrogels for tumor recurrence are summarized. The review emphasizes unique properties of hydrogels for postoperative tumor prevention and provides a broader insight into clinical transformation of hydrogels.

Qian Pu,1 Kaiyue Wang,1 Bigeng Peng,2 Kexin Chen,3 Tao Gong,4 Fu Liu,1,5 Qin Yang1 1School of Pharmacy, North Sichuan Medical College, Nanchong, 637000, People’s Republic of China; 2Oncology Department of Affiliated Hospital of North Sichuan Medical College, Nanchong, 637000, People’s Republic of China; 3Urban Vocational College of Sichuan, Meishan, 620000, People’s Republic of China; 4Key Laboratory of Drug-Targeting and Drug Delivery System of the Education Ministry and Sichuan Province, Sichuan Engineering Laboratory for Plant-Sourced Drug and Sichuan Research Center for Drug Precision Industrial Technology, West China School of Pharmacy, Sichuan University, Chengdu, 610041, People’s Republic of China; 5Department of Pharmacy, Affiliated Hospital of North Sichuan Medical College, Nanchong, 637000, People’s Republic of ChinaCorrespondence: Fu Liu, Department of Pharmacy, the Affiliated Hospital of Noth Sichuan Medical College, Nanchong, Sichuan, China, Tel +868172396935, Email nclf91@163.com Qin Yang, School of Pharmacy, North Sichuan Medical College, Nanchong, Sichuan, China, Tel +868173373323, Email qinyang201@163.comBackground: Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by arthrocele, cartilage damage and disability. Although several anti-RA drugs have been developed for long-term treatment, they require frequent local injection and lead to multiple adverse effects such as osteoporosis and myelosuppression.Purpose: Reducing the amount and frequency of anti-RA drugs methotrexate (MTX) and dexamethasone sodium phosphate (DSP) by local injection of phospholipid-based phase separation gel (PPSG) coloaded the two drugs, which presented PPSG-(+).Methods: First, We characterized PPSG-(+). And we used UV spectrophotometry and high performance liquid chromatography (HPLC) to detect drug concentration, which can clarify the drug release in vitro and in vivo, respectively. We also injected PPSG-(+) into the joint cavity of healthy rabbits to prove the safety of PPSG-(+). Then, we injected PPSG-(+) into the joint cavity of RA modeled rabbits to demonstrate the effect in anti-RA of PPSG-(+) including the thickness of joints, tumor necrosis factor (TNF)-α and interleukin (IL)-1β detection, hematoxylin–eosin (H&E) staining and computed tomography (CT) of joints.Results: Suspended particles show a tight and uniform arrangement in PPSG-(+). The gel underwent a phase transition at 20 min in vitro and 8 h in vivo, and vesicular structures reflecting its degradation and phase transition were observed in vivo. PPSG-(+) released both drugs in a sustained and fixed ratio for more than 14 days, while it proved to be safe for intra-articular injection and did not induce inflammation in a rabbit. Eventually, PPSG-(+) showed a good anti-RA effect and its potency can be maintained for 3 weeks.Conclusion: PPSG-(+) is a drug delivery system offering good biocompatibility and sustained release of MTX and DSP, leading to long-lasting anti-RA effect.Keywords: drug delivery system, in situ gel, sustained release, anti rheumatoid arthritis

Highly selective divergent oxidative dearomatization coupling reactions, in which the chemoselectivity is controlled by catalysts and bases, are reported herein. Three different kinds of polycyclic cyclohexadienones are produced from the same reactants (41 examples, 85-99% yield). Our method marks a novel copper- and palladium-catalyzed C-H oxidative dearomatization of phenolic derivatives.

A novel palladium catalyzed homodimerization of

Funder: EC | FP7 | Ideas | FP7 Ideas: European Research Council; Grant(s): 854126, The genomes of some purple photosynthetic bacteria contain a multigene puc family encoding a series of α- and β-polypeptides that together form a heterogeneous antenna of light-harvesting 2 (LH2) complexes. To unravel this complexity, we generated four sets of puc deletion mutants in Rhodopseudomonas palustris, each encoding a single type of pucBA gene pair and enabling the purification of complexes designated as PucA-LH2, PucB-LH2, PucD-LH2, and PucE-LH2. The structures of all four purified LH2 complexes were determined by cryogenic electron microscopy (cryo-EM) at resolutions ranging from 2.7 to 3.6 Å. Uniquely, each of these complexes contains a hitherto unknown polypeptide, γ, that forms an extended undulating ribbon that lies in the plane of the membrane and that encloses six of the nine LH2 αβ-subunits. The γ-subunit, which is located near to the cytoplasmic side of the complex, breaks the C9 symmetry of the LH2 complex and binds six extra bacteriochlorophylls (BChls) that enhance the 800-nm absorption of each complex. The structures show that all four complexes have two complete rings of BChls, conferring absorption bands centered at 800 and 850 nm on the PucA-LH2, PucB-LH2, and PucE-LH2 complexes, but, unusually, the PucD-LH2 antenna has only a single strong near-infared (NIR) absorption peak at 803 nm. Comparison of the cryo-EM structures of these LH2 complexes reveals altered patterns of hydrogen bonds between LH2 αβ-side chains and the bacteriochlorin rings, further emphasizing the major role that H bonds play in spectral tuning of bacterial antenna complexes.

Background Breast cancer is a malignant tumor with the highest incidence among women in the world. GREB1L is a protein coding gene. Previous studies have shown that GREB1L played an important role in lung adenocarcinoma and gastric adenocarcinoma. Currently, there is no relevant report about its role in breast cancer. Methods The Cancer Genome Atlas database was used to compare the expression level of GREB1L; TISIDB website was used for prognosis analysis; LinkedOmics database was used to predict the potential biological mechanism of GREB1L in breast cancer; Immunohistochemistry was used to detect the GREB1L expression level in breast tissue; Western blot was used to detect the GREB1L expression level in cell lines; Transwell assays, CCK8 cell proliferation assays and clone formation assays were used to detect the migration, invasion and proliferation and clone formation abilities of cells. Results GREB1L was highly expressed in breast cancer tissues and breast cancer cells; KEGG enrichment analysis suggested that GREB1L might participate in the regulation of Hedgehog signaling pathway; GREB1L affected the migration and invasion abilities of MCF7 and MDA-MB-231 cells, but not affected their proliferation and clone formation abilities. The overexpression of GREB1L in breast cancer predicted a favorable prognosis. Conclusion These results showed that GREB1L was involved in the development of breast cancer, and it may be a potential molecular marker to predict the prognosis of breast cancer.

Mussel-inspired adhesive hydrogels have been developed in biomedical fields due to their strong adhesive property, cohesive capability, biocompatibility, and hemostatic ability. Catechol-functionalized chitosan is a potential polymer used to prepare adhesive hydrogels. However, the unique gelation mechanism and self-healing properties of catechol-grafted chitosan alone have not yet been explored. Herein, catechol-grafted chitosan (CC) was synthesized and further concentrated to obtain the self-healing CC hydrogels. The gelation mechanism of CC hydrogels may be attributed to the formation of hydrogen bonding, cation–π interactions, Michael addition, or Schiff base reactions during concentration phases. Rheological studies showed that the CC hydrogel owned self-healing properties in repeated damage–healing cycles. Coherent small-angle X-ray scattering (SAXS) analyses revealed the formation of a mesoscale structure (~9 nm) as the solid content of the hydrogel increased. In situ SAXS combined with rheometry verified the strain-dependent behavior of the CC hydrogel. The CC hydrogel displayed the osmotic-responsive behavior and enhanced adhesive strength (0.38 N/cm2) after immersion in the physiological saline. The CC scaffold prepared by lyophilizing the CC hydrogel revealed a macroporous structure (~200 µm), a high swelling ratio (9656%), good compressibility, and durability. This work provides an insight into the design of using chitosan–catechol alone to produce hydrogels or scaffolds with tunable mechanical properties for further applications in biomedical fields.

Objective To provide a rare case of local recurrent Paget’s disease after nipple-sparing mastectomy (NSM) with immediate breast reconstruction with 10 years of disease-free survival and to analyze the clinical and pathological characteristics. Background Mammary Paget’s disease can be considered a rare type of local recurrence after breast cancer treatment, both in cases of conservative surgery and NSM with immediate breast reconstruction (Lohsiriwat et al, Ann Surg Oncol 19:1850-1855, 2012). Recurrent patients who present with nipple-areolar Paget’s disease usually have unfavorable primary pathological characteristics and different latency periods. However, the recurrent status in patients with favorable primary pathological characteristics and the latency periods after NSM with immediate breast reconstruction are unclear. Methods First, we present a case of local recurrent Paget’s disease in a young patient diagnosed with invasive breast carcinoma at age 30 who underwent NSM with primary silicone reconstruction. Then, the keywords “Paget’s disease” and “nipple-sparing mastectomy” were selected. Articles including the local recurrence of Paget’s disease after NSM were collected from the PubMed, Springer, and OVID databases, and the acquired relevant data were analyzed. We did not restrict our search by study design or publication date. Results Five studies describing 31 cases of local recurrent Paget’s disease after NSM with implant breast reconstruction were included. The mean patient age reported was 45 years, and the average latency period from NSM to the local recurrence of Paget’s disease was 40.2 months. Recurrent tumor histological features were Paget’s disease with ductal carcinoma in situ (DCIS) in 16 patients (50%), Paget’s disease without DCIS in 13 patients (40.6%), and Paget’s disease with ductal intraepithelial neoplasia (DIN) in 3 patients (9.4%). The primary tumor histological feature was estrogen receptor (ER)(−)/progesterone receptor (PR)(−)/human epidermal growth factor receptor (HER-2)(+) in 21 patients (77.8%). Neither locoregional relapse nor metastatic events were found in these recurrent patients who accepted NAC excision after 4–5 years of follow-up. Our reported case showed that the patient experienced pregnancy and lactation after primary adjuvant chemotherapy and endocrine therapy. However, she developed an eczematoid lesion in the NAC 120 months after breast surgery. The histopathological examination was consistent with Paget’s disease of the breast. Complete NAC and breast silicone prosthesis removal were performed. The patient accepted no systematic or local therapy and is currently alive. It is noteworthy that the biological features of the primary tumor were ER(+), PR(+), and HER-2(−); however, the recurrent tumor changed to ER(−), PR(−), and HER-2(+). Conclusions The local recurrence of Paget’s disease after NSM is uncommon; it may develop at a very early age and have a very long time to recurrence, as in our patient, who presented with recurrence 10 years after primary surgery. Surgeons should be wary of local recurrence of the nipple-areola complex after NSM in patients with ER-negative and HER-2-positive primary tumors. However, patients with ER/PR-positive and HER-2-negative tumors should not be neglected; we reported a case of an ER/PR-positive and HER-2-negative primary tumor, and ER-positive recurrent cases have the longest latency period. The local recurrence rate of Paget’s disease after NSM is low, and the prognosis is good in recurrent patients who accept further extensive NAC excision. Further systematic treatment was not considered for this patient.

The self-assembly of mechanoluminochromic polynuclear gold(I) complexes has attracted more and more attention in the field of supramolecular gold(I) chemistry. In this work, we adopted a stepwise self-assembly strategy to precisely synthesize two polynuclear gold(I) supramolecular clusters. Through cooperative Au

Saponins are kinds of antifungal compounds produced by Panax notoginseng to resist invasion by pathogens. Ilyonectria mors-panacis G3B was the dominant pathogen inducing root rot of P. notoginseng, and the abilities to detoxify saponins were the key to infect P. notoginseng successfully. To research the molecular mechanisms of detoxifying saponins in I. mors-panacis G3B, we used high-throughput RNA-Seq to identify 557 and 1519 differential expression genes (DEGs) in I. mors-panacis G3B with saponins treatments for 4H (Hours) and 12H (Hours) compared with no saponins treatments, respectively. Among these DEGs, we found 93 genes which were simultaneously highly expressed in I. mors-panacis G3B with saponins treatments for 4H and 12H, they mainly belong to genes encoding transporters, glycoside hydrolases, oxidation–reduction enzymes, transcription factors and so on. In addition, there were 21 putative PHI (Pathogen–Host Interaction) genes out of those 93 up-regulated genes. In this report, we analyzed virulence-associated genes in I. mors-panacis G3B which may be related to detoxifying saponins to infect P. notoginseng successfully. They provided an excellent starting point for in-depth study on pathogenicity of I. mors-panacis G3B and developed appropriate root rot disease management strategies in the future.

A polyphosphate-producing bacterium, YG09T, was isolated from the rhizosphere of Salvia miltiorrhiza. Its colonies were 2.0–3.0 mm in diameter, smooth, circular, convex and yellow after growth on R2A at 28 °C for 72 h, with aerobic, Gram-stain-negative, non-motile and rod-shaped bacteria. The strain was found to grow at 10–40 °C (optimum 37 °C), pH 5.5–8.0 (optimum 6.0), with 0–0.6% (w/v) NaCl (optimum 0). Chemotaxonomic analysis showed menaquinone-7 as the only quinone present; C15: 1 iso G, C15: 1 iso, C16: 0, C16: 0 3OH, C17: 0 iso 3OH, summed feature 3 (C16:1 ω7c and/or C16:1 ω6c) as the major fatty acids (> 5%), and phosphatidylethanolamine, three unidentified phospholipids, four unidentified polar lipids, three unidentified aminolipids, and one unidentified amino phospholipid as the polar lipids. The DNA G + C content was 44.6 mol%. The 16S rRNA gene sequences of the isolate showed highest similarities to Panacibacter ginsenosidivorans Gsoil 1550T (93.6%), Filimonas endophytica SR2-06T (93.4%), Parasegetibacter terrae SGM2-10T (92.8%), and Arvibacter flaviflagrans C-1-16T (92.7%), within the family Chitinophagaceae of the phylum Bacteroidetes. The ANI values between strain YG09T and Panacibacter ginsenosidivoran Gsoil 1550T, Filimonas endophytica SR2-06T and Filimonas lacunae YT21T were 69.4, 68.3 and 68.7%, respectively. Based on phenotypic, genotypic and phylogenetic analyses, strain YG09T represents a novel genus in the family Chitinophagaceae, for which the name Foetidibacter luteolus gen. sp. nov. is proposed. The type strain is Foetidibacter luteolus YG09T (= MCCC 1K04042T = KCTC 72595T).

The identity of the floral organs is defined by a small group of transcriptional regulators, and the activities of these proteins can specify the feature of the different whorls in flowers. In the last thirty years, formation of the floral organs remained as the main research subject in plant, especially in Arabidopsis thaliana. By using A. thaliana as material, relevant research works have established a basic architecture for the development of floral organs in higher plants, and it was named as the ABCDE model. In accordance with this model, the identity of the different floral organs can be confirmed by the specific combinations of A-, B-, C-, D-, or E-class floral homeotic genes. SEP-like genes encode MADS transcription factors required for the development of all the four whorls of floral organs and for the determinacy of the floral meristems. In ABCDE model, these genes are defined as E-class floral homeotic genes. A great deal of studies have shown that the proteins encoding by these genes are the main regulators of flower development and act pivotal parts not only in organ identification but also in organ morphogenesis. In this paper, the functions of the E-class floral homeotic genes in dicotyledons are reviewed, and the progress that has been made in characterization of the floral organ identity factors in A. thaliana and other dicotyledons is discussed.

E-class MADS-box genes, SEPALLATA (SEP), participate in various aspects of plant development together with B-, C- and D-class MADS-box genes. IiSEP4, a homologous gene of SEP4, was cloned from Isatis indigotica. IiSEP4 was highly expressed in sepals, and its mRNA was mildly detected in leaves, inflorescences, flowers, stamens and young silicles. Constitutive expression of IiSEP4 in Arabidopsis thaliana caused early flowering, accompanied by the reduction of flowers and floral organs. Moreover, the sepals in some flowers were transformed into carpelloid structures with stigmatic papillae, and obviously accompanied by ovule formation. Yeast two-hybrid assays demonstrated that IiSEP4 interacts with other woad MADS proteins to determine the identity of floral organs. These findings reveal the important roles of IiSEP4 in floral development of I. indigotica. The results of this study can lay a foundation for further study on biological functions of MADS transcriptional factors in I. indigotica.

Background P53 pathway inactivation plays an important role in the process of breast cancer tumorigenesis. Post-translational protein modification abnormalities have been confirmed to be an important mechanism underlying inactivation of p53. Numerous deubiquitinating enzymes are aberrantly expressed in breast cancer, and a few deubiquitination enzymes can deubiquitinate and stabilize p53. Here, we report that ovarian tumor (OTU) deubiquitinase 3 (OTUD3) is a deubiquitylase of p53 in breast carcinoma (BC). Methods Correlations between the mRNA expression levels of OTUD3, TP53 and PTEN and the prognosis of BC were assessed with the Kaplan-Meier Plotter tool. OTUD3 protein expression in 80 pairs of specimens in our cohort was examined by immunohistochemistry and western blotting. The relationship among OTUD3, p53, and p21 proteins was analyzed. Half-life analysis and ubiquitylation assay were performed to elucidate the molecular mechanism by which OTUD3 stabilizes p53. The interaction between OTUD3 and p53 in BC cells was verified by a co-immunoprecipitation assay and GST pulldown experiments. MTS assay for proliferation detection, detection of apoptosis induced by cisplatin and colony formation assay were employed to investigate the functional effects of OTUD3 on breast cancer cells. Results OTUD3 downregulation is correlated with a poor prognosis in BC patients. OTUD3 expression is decreased in breast cancer tissues and not associated with the histological grade. OTUD3 also inhibits cell proliferation and clone formation and increases the sensitivity of BC cells to apoptosis induced by chemotherapy drugs. Reduced OTUD3 expression accompanied by decreased p53 abundance is correlated with human breast cancer progression. Ectopic expression of wild-type OTUD3, but not its catalytically inactive mutant, stabilizes and activates p53. Mechanistically, OTUD3 interacts directly with p53 through the amino-terminal OTU region. Finally, OTUD3 protects p53 from murine double minute 2 (Mdm2)-mediated ubiquitination and degradation, enabling the deubiquitination of p53 in BC cells. Conclusions In summary, we found that OTUD3 may be a potential therapeutic target for restoring p53 function in breast cancer cells and suggest that the OTUD3-p53 signaling axis may play a critical role in tumor suppression.

A urea-utilizing bacterium, designated Q2-2T, was isolated from landfill. Cells of strain Q2-2T were Gramstain-negative, aerobic, short-rod bacteria. Strain Q2-2T was observed to grow at a temperature range of 15-37℃ (optimum 30℃), a pH range of 5.5-9.5 (optimum pH 8.0) and 0-4% (w/v) NaCl (optimum 1%). The major respiratory quinone detected was Q-8, and the polar lipids identified were mainly diphosphatidyl glycerol, phosphatidylethanolamine, phosphatidylmethylethanolamine, and phosphatidyl glycerol. Based on the 16S rRNA gene sequence, strain Q2-2T had the highest similarity with Paracandidimonas caeni 24T (97.99%), Pusillimonas soli MJ07T (97.54%), Parapusillimonas granuli Ch07T (97.27%), Pusillimonas ginsengisoli DCY25T (97.09%) and Paracandidimonas soli IMT-305T (96.39%). The ANI values between strain Q2-2T and the above related type strains were 71.02%, 73.52%, 74.32%, 74.59% and 72.29%, respectively. The DNA G + C content of strain Q2-2T was 61.1%. Therefore, strain Q2-2T represents a novel species of the genus Paracandidimonas, for which the name Paracandidimonas lacteus sp. nov. (type strain Q2-2T= CGMCC 1.19179T =JCM 34906T) is proposed.

A urea-utilizing bacterium, designated Q2-2

Reaction centre light-harvesting 1 (RC-LH1) complexes are the essential components of bacterial photosynthesis. The membrane-intrinsic LH1 complex absorbs light and the energy migrates to an enclosed RC where a succession of electron and proton transfers conserves the energy as a quinol, which is exported to the cytochrome bc1 complex. In some RC-LH1 variants quinols can diffuse through small pores in a fully circular, 16-subunit LH1 ring, while in others missing LH1 subunits create a gap for quinol export. We used cryogenic electron microscopy to obtain a 2.5 Å resolution structure of one such RC-LH1, a monomeric complex from Rhodobacter sphaeroides. The structure shows that the RC is partly enclosed by a 14-subunit LH1 ring in which each αβ heterodimer binds two bacteriochlorophylls and, unusually for currently reported complexes, two carotenoids rather than one. Although the extra carotenoids confer an advantage in terms of photoprotection and light harvesting, they could impede passage of quinones through small, transient pores in the LH1 ring, necessitating a mechanism to create a dedicated quinone channel. The structure shows that two transmembrane proteins play a part in stabilising an open ring structure; one of these components, the PufX polypeptide, is augmented by a hitherto undescribed protein subunit we designate as protein-Y, which lies against the transmembrane regions of the thirteenth and fourteenth LH1α polypeptides. Protein-Y prevents LH1 subunits 11-14 adjacent to the RC QB site from bending inwards towards the RC and, with PufX preventing complete encirclement of the RC, this pair of polypeptides ensures unhindered quinone diffusion.

The reaction centre light-harvesting 1 (RC-LH1) complex is the core functional component of bacterial photosynthesis. We determined the cryo-electron microscopy (cryo-EM) structure of the RC-LH1 complex from Rhodospirillum rubrum at 2.5 Å resolution, which reveals a unique monomeric bacteriochlorophyll with a phospholipid ligand in the gap between the RC and LH1 complexes. The LH1 complex comprises a circular array of 16 αβ-polypeptide subunits that completely surrounds the RC, with a preferential binding site for a quinone, designated QP, on the inner face of the encircling LH1 complex. Quinols, initially generated at the RC QB site, are proposed to transiently occupy the QP site prior to traversing the LH1 barrier and diffusing to the cytochrome bc1 complex. Thus, the QP site, which is analogous to other such sites in recent cryo-EM structures of RC-LH1 complexes, likely reflects a general mechanism for exporting quinols from the RC-LH1 complex.

Background: To obtain a thorough comprehension of the profile and prognosis of activating transcription factor (ATF) family members in breast cancer.Method: We searched Oncomine, GEPIA, cBioPortal, Kaplan-Meier plotter, and CancerSEA databases to assess expression level, prognostic value, and functions of ATFs in breast cancer. Results: In breast cancer, we found that the expression levels of genes like ATF1, ATF5, and ATF6, were higher than in normal tissues. While the expression levels of ATF3, ATF4, ATF7 were lower in the former than in the latter. Similarly, the ATFs protein expressions were consistent with this in the Human Protein Atlas database. High expressions of ATF2, ATF4, and ATF6-7 were associated with good relapse-free survival. Increased expressions of ATF4 and ATF7 had high overall survival. Conversely, the mRNA expression of ATF1 was negatively correlated with distant metastasis-free survival. Similarly, high expression of ATF2 had reduced post-progression survival. Conclusions: ATF1 was a target of potential therapeutic interest for breast cancer, and ATF4 and ATF6-7 were potential prognostic factors in evaluating breast cancer.

The silk gland is characterized by high protein synthesis. However, the molecular mechanisms controlling silk gland growth and silk protein synthesis remain undetermined. Here we demonstrated that CRISPR/Cas9-based knockdown of let-7 or the whole cluster promoted endoreduplication and enlargement of the silk gland, accompanied by changing silk yield, whereas transgenic overexpression of let-7 led to atrophy and degeneration of the silk gland. Mechanistically, let-7 controls cell growth in the silk gland through coordinating nutrient metabolism processes and energy signalling pathways. Transgenic overexpression of pyruvate carboxylase, a novel target of let-7, resulted in enlargement of the silk glands, which is consistent with the abnormal phenotype of the let-7 knockdown. Overall, our data reveal a previously unknown miRNA-mediated regulation of silk gland growth and physiology and shed light on involvement of let-7 as a critical stabilizer and booster in carbohydrate metabolism, which may have important implications for understanding of the molecular mechanism and physiological function of specialized organs in other species.

In order to ascertain the regulatory mechanism of fruit development in Isatis indigotica Fortune, the complementary DNA (cDNA) sequence of the SHATTERPROOF 2 (SHP2) orthologous gene was identified by Rapid Amplification of cDNA Ends technology and the corresponding gene was named IiSHP2. The expression pattern of IiSHP2 was determined by quantitative reverse transcription-polymerase chain reaction and wild-type Col-0 Arabidopsis plants were transformed with the IiSHP2 gene using Agrobacterium tumefaciens and the floral-dip method. Expression analyses indicated that IiSHP2 was highly expressed in flowers, silicles and seeds. Compared to wild-type plants, IiSHP2 transgenic lines bolted earlier. Detailed phenotypic observations showed that the size of the rosette and cauline leaves in transgenic lines was reduced and the cauline leaves of the transgenic lines were incurved and displayed a funnel-like shape. During the reproductive growth stage, IiSHP2 transgenic plants produced shortened sepals and the flower buds were not encapsulated completely. Moreover, the petals of the transgenic lines were converted into stamineous tissues, accompanied by exposed stamens, short malformed siliques and wrinkled valves, indicating a severe decline in fertility. These experimental conclusions are valuable as a reference for the breeding of medicinal plants.

Speed profile design and tracking control are two important issues to ensure the performance of the automatic train operation system. Aiming to realize the maximum optimization probability of train running, this paper constructs an integrated optimal method through speed profile design and fuzzy PID controller design. In the recommended speed profile design, three typical running strategies are included in the proposed comprehensive scheme to extend solution space, and the constructed train performance simulation is combined with NSGA-II algorithm to solve such multiobjective problem. As for recommended speed profile tracking, in order to improve the tracking performance of the train, which is a nonlinear system, a fuzzy PID controller is designed to adaptively adjust PID gains. Note that multiple indicators of punctually, energy consumption, stopping accuracy and comfort are all considered in the integrated design. Taking Beijing Subway Line 8 as the numerical example, the results show that the design of speed profile and fuzzy PID controller are both effective, and energy consumption saves about 10.40% with other better indicators compared with the original data.

BackgroundFemaleAedes aegyptimosquitoes are vectors of arboviruses that cause diverse diseases of public health significance. Blood protein digestion by midgut proteases provides anautogenous mosquitoes with the nutrients essential for oocyte maturation and egg production. Midgut-specific miR-1174 affects the functions of the midgut through its target geneserine hydroxymethyltransferase(SHMT). However, less is known aboutSHMT-regulated processes in blood digestion by mosquitoes.MethodsRNAi ofSHMTwas realized by injection of the double-stranded RNA at 16 h post-eclosion. The expression ofSHMTat mRNA level and protein level was assayed by real-time PCR and Western blotting, respectively. Statistical analyses were performed with GraphPad7 using Student’s t-test.ResultsHere, we confirmed that digestion of blood was inhibited inSHMTRNAi-silenced femaleA. aegyptimosquitoes. Evidence is also presented that allSHMT-depleted female mosquitoes lost their flight ability and died within 48 h of a blood meal. Furthermore, most examined digestive enzymes responded differently in their transcriptional expression to RNAi depletion ofSHMT, with some downregulated, some upregulated and some remaining stable. Phylogenetic analysis showed that transcriptional expression responses toSHMTsilence were largely unrelated to the sequence similarity between these enzymes.ConclusionsOverall, this research shows thatSHMTwas expressed at a low level in the midgut ofAedes aegyptimosquitoes, but blood-meal digestion was inhibited whenSHMTwas silenced. Transcriptional expressions of different digestive enzymes were affected in response toSHMTdepletion, suggesting thatSHMTis required for the blood-meal digestion in the midgut and targetingSHMTcould provide an effective strategy for vector mosquito population control.

Besides energy-efficiency, people also want train operation to be comfortable, punctual and parking precise. In this paper, a multi-objective model for automatic train operation in urban railway is proposed by unifying dimensions of different objectives firstly. This model is built by applying multi-objective decision with the penalty function, based on the analysis of train performance and its operation environment. Then a genetic algorithm is developed to solve this model and obtain the optimal recommended speed profiles. Thirdly, fuzzy controller is designed to achieve track recommended speed profiles. Finally, with the help of Matlab software, control effect is verified based on simulation. From the simulation results, it can be seen this strategy can meet the requirement of multi-objective, which are energy-saving, parking precisely, running punctually and comfort.

The N6-methyladenosine (m6A) has been considered as a new layer of epitranscriptomic regulation on mRNA processing, stability, and translation. However, potential roles of m6A RNA methylation modification in tumor immune microenvironment (TIME) of breast cancer are yet fully understood. In this study, we comprehensively evaluated the genetic variations and transcript expressions of 15 m6A regulators in 1,079 breast cancer samples from the Cancer Genome Atlas (TCGA) database. We validated major regulators had significantly differential mRNA and protein expression in tumor tissue compared to normal tissues from 39 pairs of clinical breast cancer samples with different molecular subtypes, and especially high expression of m6A readers YTHDF1 and YTHDF3 predicted poor survival. Two clusters of breast cancer patients identified by the 15 m6A regulators’ pattern showed distinct overall survival, immune activation status, and immune cell infiltration, and clinical samples confirmed the diversity of lymphocytic infiltration. The profiles of these two clusters accorded with that of two classical cancer-immune phenotypes, immune-excluded and immune-inflamed phenotypes, it suggested that m6A regulators-based patterns might serve as crucial mediators of TIME in breast cancer. Moreover, the m6A phenotype-related gene signatures could also be survival predictor in breast cancer. Therefore, comprehensive evaluation of tumor m6A modification pattern will contribute to enhance our understanding of the characterization of immune cell infiltration in the tumor microenvironment and promote the responsiveness of breast cancer to immunotherapy.

Background: The N6-methyladenosine (m6A) has been considered as a new layer of epitranscriptomic regulation on mRNA processing, stability and translation. However, potential roles of m6A RNA methylation modification in tumor immune microenvironment (TIME) of breast cancer are yet fully understood.Methods: We comprehensively evaluated the genetic variations and transcript expressions of 15 m6A regulators, and clinicopathological features in 1,079 breast cancer samples from The Cancer Genome Atlas (TCGA) database. The mRNA and protein levels of several m6A regulators were validated by RT-qPCR, western blot and immunohistochemistry staining in clinical samples from 39 patients with breast cancer. The prognostic values of m6A regulators were systematically evaluated in different database. We correlated the m6A modification patterns of breast cancer with the immune microenvironment and cancer-immune phenotypes. The m6A regulators-related gene signatures were also analyzed to predict the survival of patients.Results: Some m6A regulators’ CNV events might be potential biomarkers for patient’s stage and prognosis in breast cancer. Major regulators had significantly differential mRNA and protein expression in tumor tissue compared to normal samples among different molecular subtypes of breast cancer, and especially high expression of m6A readers YTHDF1 and YTHDF3 predicted poor survival. Two clusters of breast cancer patients identified by the 15 m6A regulators’ pattern showed distinct overall survival, immune activation status and immune cell infiltration. The profiles of these two clusters accorded with that of two classical cancer-immune phenotypes, immune-excluded and immune-inflamed phenotypes. Moreover, the m6A phenotype-related gene signatures could also be survival predictor in breast cancer.Conclusions: The m6A regulators-based patterns might serve as crucial mediators of TIME in breast cancer. Comprehensive evaluation of tumor m6A modification pattern will contribute to enhance our understanding of the characterization of immune cell infiltration in the tumor microenvironment and promote the responsiveness of breast cancer to immunotherapy.

Mesenchymal stem cell derived (MSC) exosomes are extracellular vesicles with a diameter of about 50-200 nm. Exosomes contain a large number of biologically active substances including mRNAs, miRNAs, cytokines, chemokines, proteins, lipids, etc. MSC exosomes exert biological effects through paracrine and endocrine pathways in vivo. Uncontrolled inflammation and multiple organ dysfunction are the key roles in the progression of sepsis, moreover, heart, lungs, kidneys and brain are the general target organs to be damaged. MSC exosomes regulate the expression of cytokines, the production of inflammatory cell, the levels of inflammatory response and the recovery of damaged tissues or organ function. Therefore, studying the application of MSC exosomes is significant for the clinical treatment of sepsis. This article reviews the systemic inflammation regulation by MSC exosomes and its protective function on targeted organs such as heart, lungs, kidneys, brain, liver, etc. to provide evidences for the treatment of sepsis.

With the increasing demand for high quality and environmentally safe or green food, Biological Control Agents (BCAs) are playing critical roles in green agriculture, which in turn has paved the way for the requirement of effective, appropriate microbial antagonists. In this study, Mucor moelleri AA1 was isolated and investigated for its growth promotion and antagonism against Athelia rolfsii and Colletotrichum gloeosporiodes. The results showed a high antagonistic activity of M. moelleri against A. rolfsii and C. gloeosporiodes with percentage inhibitions of 73 % and 86 % respectively using the dual plate method, and the same antagonistic activity was also observed in liquid cocultures. A pot study analysis showed significant suppression of the diseases as well as growth promotion on tomato. Scanning electron microscopy (SEM) indicated that M. moelleri inhibited the growth of mycelium and the production of web-like materials. Based on headspace-solid phase microextraction (HS-SPME) analysis, microbial volatile compounds were determined, which were mainly aromatic compounds and alkaloids. Also, several antagonistic enzymes, such as β-1, 3- glucanase, proteases, catalase and ACC deaminase as well as the phytohormone IAA, were found to be produced by M. moelleri. Overall, these results combine to make M. moelleri a good prospective candidate for biological control and as a plant growth-promoting agent. The present study appears to be the first report identifying M. moelleri as a biological control agent.

Saponins are kinds of antifungal compounds produced by P. notoginseng to resist invasion by pathogens. I. mors-panacis G3B was the dominant pathogen inducing root rot of P. notoginseng, and the abilities to detoxify saponins were the key to infect P. notoginseng successfully. To research the molecular mechanisms of detoxifying saponins in I. mors-panacis G3B, we used high-throughput RNA-Seq to identify 557 and 1519 differential expression genes (DEGs) in I. mors-panacis G3B with saponins treatments for 4 H and 12 H compared with no saponins treatments, respectively. Among these DEGs, we found 93 genes which were simultaneously highly expressed in I. mors-panacis G3B with saponins treatments for 4 H and 12 H, they mainly belong to genes encoding transporters, glycoside hydrolases, oxidation-reduction enzymes, transcription factors and so on. In addition, there were 21 putative PHI (Pathogen-Host Interaction) genes out of those 93 up-regulated genes. In this report, we identified virulence associated genes in I. mors-panacis G3B which may be related to detoxifying saponins to infect P. notoginseng successfully. They provided an excellent starting point for in-depth study on pathogenicity of I. mors-panacis G3B and developed appropriate root rot disease management strategies in the future.

Background: P53 pathway inactivation plays an important role in the process of breast cancer tumorigenesis. Post-translational protein modification abnormalities have been confirmed to be an important mechanism underlying inactivation of p53. Numerous deubiquitinating enzymes are aberrantly expressed in breast cancer, and a few deubiquitination enzymes can deubiquitinate and stabilize p53. Here, we report that ovarian tumor (OTU) deubiquitinase 3 (OTUD3) is a deubiquitylase of p53 in breast carcinoma (BC).Methods: Correlations between the mRNA expression levels of OTUD3, TP53 and PTEN and the prognosis of BC were assessed with the Kaplan-Meier Plotter tool. OTUD3 protein expression in 80 pairs of specimens in our cohort was examined by immunohistochemistry and western blotting. The relationship among OTUD3, p53, and p21 proteins was analyzed. Half-life analysis and ubiquitylation assay were performed to elucidate the molecular mechanism by which OTUD3 stabilizes p53. The interaction between OTUD3 and p53 in BC cells was verified by a co-immunoprecipitation assay and GST pulldown experiments. MTS assay for proliferation detection, detection of apoptosis induced by cisplatin and colony formation assay were employed to investigate the functional effects of OTUD3 on breast cancer cells. Results: OTUD3 downregulation is correlated with a poor prognosis in BC patients. OTUD3 expression is decreased in breast cancer tissues and not associated with the histological grade. OTUD3 also inhibits cell proliferation and clone formation and increases the sensitivity of BC cells to apoptosis induced by chemotherapy drugs. Reduced OTUD3 expression accompanied by decreased p53 abundance is correlated with human breast cancer progression. Ectopic expression of wild-type OTUD3, but not its catalytically inactive mutant, stabilizes and activates p53. Mechanistically, OTUD3 interacts directly with p53 through the amino-terminal OTU region. Finally, OTUD3 protects p53 from murine double minute 2 (Mdm2)-mediated ubiquitination and degradation, enabling the deubiquitination of p53 in BC cells. Conclusions: In summary, we found that OTUD3 may be a potential therapeutic target for restoring p53 function in breast cancer cells and suggest that the OTUD3-p53 signaling axis may play a critical role in tumor suppression.

Background The architecture of inflorescence and the development of floral organs can influence the yield of seeds and have a significant impact on plant propagation. E-class floral homeotic MADS-box genes exhibit important roles in regulation of floral transition and differentiation of floral organs. Woad (Isatis indigotica) possesses unique inflorescence, floral organs and fruit. However, very little research has been carried out to determine the function of MADS-box genes in this medicinal cruciferous plant species. Results SEPALLATA orthologs in I. indigotica were cloned by degenerate PCR. The sequence possessing the highest identity with SEP2 and SEP4 of Arabidopsis were named as IiSEP2 and IiSEP4, respectively. Constitutive expression of IiSEP2 in Columbia (Col-0) ecotype of Arabidopsis led to early flowering, and the number of the flowers and the lateral branches was reduced, indicating an alteration in architecture of the inflorescences. Moreover, the number of the floral organs was declined, the sepals were turned into carpelloid tissues bearing stigmatic papillae and ovules, and secondary flower could be produced in apetalous terminal flowers. In 35S::IiSEP4-GFP transgenic Arabidopsis plants in Landsberg erecta (Ler) genetic background, the number of the floral organs was decreased, sepals were converted into curly carpelloid structures, accompanied by generation of ovules. Simultaneously, the size of petals, stamens and siliques was diminished. In 35S::IiSEP4-GFP transgenic plants of apetalous ap1 cal double mutant in Ler genetic background, the cauliflower phenotype was attenuated significantly, and the petal formation could be rescued. Occasionally, chimeric organs composed of petaloid and sepaloid tissues, or petaloid and stamineous tissues, were produced in IiSEP4 transgenic plants of apl cal double mutant. It suggested that overexpression of IiSEP4 could restore the capacity in petal differentiation. Silencing of IiSEP4 by Virus-Induced Gene Silencing (VIGS) can delay the flowering time, and reduce the number and size of the floral organs in woad flowers. Conclusion All the results showed that SEPALLATA-like genes could influence the architecture of the inflorescence and the determinacy of the floral meristems, and was also related to development of the floral organs.

Objective To investigate the effects of necroptosis induced by vibrio vulnificus cytolysin (VVc) on immortalized bone marrow-derived macrophage cell line (iBMDM) and its mechanism. Methods The iBMDM were divided into five groups, control group, VVc(2μg/ml) group, VVc(4μg/ml) group, Necrostatin-1 (Nec-1)+VVc(2μg/ml) group, Nec-1+VVc(4μg/ml) group based on the concentration of VVc and whether the cells received receptor interacting protein 1 (RIP1). In the RIP1 inhibited groups, iBMDM was pretreated with Nec-1 (5μmol/L) before stimulation with VVc. Cells and culture supernatant were harvested at 2h after the challenge by cytolysin. Lactate dehydrogenase (LDH) activity in cell-culture supernatant following VVc stimulation was determined to evaluate cell death. The cell state was detected by flow cytometry with Annexin V/propidium iodide staining, and the expression of phosphorylated mixed lineage kinase domain-like (MLKL) and pyroptosis- and necroptosis-related proteins (caspase-1, caspase-11, gasdermin D) were detected by Western blotting. Results The LDH level in the culture supernatant was increased in the groups stimulated by VVc and could also be inhibited by pretreatment with Nec-1 (P＜0.05). Compared with control and RIP1- inhibited groups, the VVc-treatment induced a high amount of the double positive staining cells in the VVc group (P＜0.05). In the RIP1 inhibited groups, Nec-1 pretreatment could significantly decrease the levels of pMLKL (P＜0.05), compared with VVc group. None of the cleaved pyroptosis-related protein was detected by Western blotting. Conclusion Necroptosis induced by RIP1 may play an important role in VVc injury model of iBMDM, furthermore, Nec-1 could reduce the degree of injury after VVc challenge. DOI: 10.11855/j.issn.0577-7402.2018.05.11

PurposeThis paper aims to provide an optimization method of workflow for publishing houses and electronic book (e-book) studies in the field of digital publishing.Design/methodology/approachBased on the studies of publishing houses in Beijing, the present conversion workflow is illustrated using a functional modeling methodology. Then, the workflow is analyzed using 5W1H (why, who, what, where, when, how) methodology and optimized using ECRSI (eliminate, combine, rearrange, simplify and increase) principles. To validate the optimization effect, the workflow before and after optimization are generated and implemented by the ExtendSim® simulation software.FindingsThe simulation results show that under similar circumstances, both quantity and quality of the products are improved after optimization, which indicate that the optimization method is effective.Practical implicationsElectronic PUBlication (EPUB) has significant requirements to satisfy the needs of the mobile reading market and to earn increased profits, whereas some e-books are still preserved in a portable document format (PDF). This study results in the enhanced EPUB quality and production efficiency of the PDF-to-EPUB format conversion workflow in publishing houses. Publishing houses around the world can refer to this study to make a similar optimization when handling PDF-to-EPUB.Originality/valueThis research introduces the traditional industrial engineering analytical techniques to the workflow optimization of e-book conversion. Compared with the most of other methods used to optimize workflow, this method is simpler, more efficient and more suitable for e-book format conversion.

Intracytoplasmic vesicles (chromatophores) in the photosynthetic bacterium Rhodobacter sphaeroides represent a minimal structural and functional unit for absorbing photons and utilising their energy for the generation of ATP. The cytochrome bc1 complex (cytbc1) is one of the four major components of the chromatophore alongside the reaction centre-light harvesting 1-PufX core complex (RC-LH1-PufX), the light-harvesting 2 complex (LH2), and ATP synthase. Although the membrane organisation of these complexes is known, their local lipid environments have not been investigated. Here we utilise poly(styrene-alt-maleic acid) (SMA) co-polymers as a tool to simultaneously determine the local lipid environments of the RC-LH1-PufX, LH2 and cytbc1 complexes. SMA has previously been reported to effectively solubilise complexes in lipid-rich membrane regions whilst leaving lipid-poor ordered protein arrays intact. Here we show that SMA solubilises cytbc1 complexes with an efficiency of nearly 70%, whereas solubilisation of RC-LH1-PufX and LH2 was only 10% and 22% respectively. This high susceptibility of cytbc1 to SMA solubilisation is consistent with this complex residing in a locally lipid-rich region. SMA solubilised cytbc1 complexes retain their native dimeric structure and co-purify with 56 ± 6 phospholipids from the chromatophore membrane. We extended this approach to the model cyanobacterium Synechocystis sp. PCC 6803, and show that the cytochrome b6f complex (cytb6f) and Photosystem II (PSII) complexes are susceptible to SMA solubilisation, suggesting they also reside in lipid-rich environments. Thus, lipid-rich membrane regions could be a general requirement for cytbc1/cytb6f complexes, providing a favourable local solvent to promote rapid quinol/quinone binding and release at the Q0 and Qi sites., Graphical abstract Image 1, Highlights • SMA preferentially solubilises cytbc1 from chromatophore membranes. • Solubilised cytbc1 SMALPs contain dimeric complexes co-purified with 56 lipids. • SMA-resistant fractions contain RC-LH1-PufX and LH2 rich membrane patches. • The Rba. sphaeroides cytbc1 complex is likely to reside in a lipid-rich environment. • Similar results for Synechocystis suggest cytbc1/b6f may be universally lipid-rich.

When the power source of a voltage source converter (VSC) station at the sending end solely depends on wind power generation, the station is operating in an islanding mode. In this case, the power fluctuation of the wind power will be entirely transmitted to the receiving-end grid. A self-regulation scheme of power fluctuation is proposed in this paper to solve this problem. Firstly, we investigated the short-time variability characteristic of the wind power in a multi-terminal direct-current (MTDC) project in China. Then we designed a virtual frequency (VF) control strategy at the VSC station based on the common constant voltage constant frequency (CVCF) control of VSC station. By cooperating with the primary frequency regulation (PFR) control at the wind farms, the self-regulation of active power pooling at the VSC station was realized. The control parameters of VF and PFR control were carefully settled through the steady-state analysis of the MTDC grid. The self-regulation effect had been demonstrated by a twenty-four-hour simulation. The results showed that the proposed scheme could effectively smoothen the power fluctuation.