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Summary The lack of model single-atom catalysts (SACs) and atomic-resolution operando spectroscopic techniques greatly limits our comprehension of the nature of catalysis. Herein, based on the designed model single-Fe-atom catalysts with well-controlled microenvironments, we have explored the exact structure of catalytic centers and provided insights into a spin-crossover-involved mechanism for oxygen reduction reaction (ORR) using operando Raman, X-ray absorption spectroscopies, and the developed operando 57Fe Mossbauer spectroscopy. In combination with theoretical studies, the N-FeN4C10 moiety is evidenced as a more active site for ORR. Moreover, the potential-relevant dynamic cycles of both geometric structure and electronic configuration of reactive single-Fe-atom moieties are evidenced via capturing the peroxido (∗O2−) and hydroxyl (∗OH−) intermediates under in situ ORR conditions. We anticipate that the integration of operando techniques and SACs in this work shall shed some light on the electronic-level insight into the catalytic centers and underlying reaction mechanism.

Fuzhuan brick tea (FBT) is an indigenous dark tea fermented by fungal and bacterial communities, and the special favor of FBT attributes to the actions of microbiota, while the correlation between bacteria and fungi attracts little attention. In the present study, therefore, the compositions of microbial communities in 10 samples of FBT from four tea processing factories were evaluated by Illumina MiSeq sequencing. Bacterial taxonomy analysis revealed that the phylum Firmicutes and Proteobacteria were the predominant bacteria in all samples. Fungal taxonomy analysis revealed that the phylum Ascomycota, family Trichocomaceae, genus Aspergillus was the dominant fungus in all samples. Moreover, Aspergillus (OTU1, OTU2, OTU16, OTU30, OTU31 and OTU37), Wallemia (OTU3), Penicillium (OTU32), Candida (OTU33, OTU35, OTU40 and OTU43), Debaryomyces (OTU48) and Rhodotorula (OTU68) showed higher correlation with bacteria in FBT. Furthermore, a strain Aspergillus cristatus was isolated from FBT, and its morphology and molecular taxonomy were observed and analyzed by scanning electron microscopy and sequencing. Hence, this is the first report to reveal the correlation between bacterial and fungal communities in FBT, and A. cristatus may have potential application to improve the quality of FBT.

Ischemic stroke is among the leading causes of death and disability across the globe. Post-stroke neuroinflammation contributes to the pathophysiology of ischemic stroke in the acute phase through damaging neurons in the penumbra region. Infiltrating regulatory T cells (Treg cells) provide neuronal protection in ischemic brains. In the current study using a mouse-transient middle cerebral artery occlusion (MCAO) model, we characterized the changes of sirtuin expression in infiltrating Treg cells in the acute phase of ischemia. We found that Sirt2 was remarkably upregulated in infiltrating Treg cells at day 3 post-MCAO. In vitro inhibition of Sirt2 activity enhanced the expression of immunosuppression-associated molecules including forkhead box P3 (Foxp3) in Treg cells. Using a lentiviral system to express exogenous Sirt2 in Treg cells, we found that Sirt2 weakened the anti-inflammatory effect of Treg cells on pro-inflammatory macrophages. Additionally, post-MCAO microglia increased Sirt2 expression in Treg cells in a cell-to-cell contact manner. We further found that microglia remarkably induced hypoxia-inducible factor 1-alpha (HIF-1α) expression in Treg cells, and inhibition of HIF-1α abolished microglia-induced Sirt2 upregulation. Collectively, we discovered a novel mechanism by which the immunoregulatory activity of infiltrating Treg cells is modulated after ischemia.

The objective of this study was to investigate the effect of Mycosporine-like amino acids (MAAs) extracted from Porphyra tenera skin against UV irradiation-induced photoaging using an ICR mouse model of skin photoaging and to explore the curative effects of the compounds in MAAs. The skin damage and collagenous tissue impairments induced by ultraviolet radiation were observed by histopathological analysis, and the effects of MAAs on protecting against skin damage and maintaining an intact structure of collagenous tissue were studied. The expression of NF-κB and the MAPK signaling pathway and nuclear transcription factors MMP-1, MMP-3 and TNF-α was analyzed used quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) and ELISA assays. The results showed that the MAAs extracted from Porphyra tenera contained Porphyra-334 and shinorine, which could prevent skin photoaging induced by ultraviolet irradiation and reduce the damage to collagen and elastin. Meanwhile, MAAs significantly inhibited the decrease in hydroxyproline and collagen content and protected against pathological damage to collagen fibers in photoaging skin. MAAs resulted in a reduction in the expression of interstitial collagenase (MMP-1), matrix metalloproteinases (MMP-3) and tumor necrosis factor-α (TNF-α) and a reduction in the content of tissue matrix metalloproteinase (MMPs). Furthermore, MAAs may promote procollagen synthesis by downregulating the expression of TNF-α and downregulating the levels of MMPs, which demonstrates that MAAs are involved in matrix collagen synthesis by activating the NF-kB pathway in photoaging skin. Based on these results, we concluded that MAAs protect skin from UV irradiation-induced photodamage, and therefore, MAAs may be a potentially effective agent for the prevention of photoaging.

In this study, the crude exopolysaccharides (CEPSs) from fungus Aspergillus cristatus (MK346334, NCBI) isolated from Fuzhuan brick tea and its main purified fraction (EPSs-2) were investigated. Using the RAW264.7 cell model, EPSs-2 exhibited an excellent immunomodulatory effect in vitro. Then, the regulating effects of EPSs on immune function and gut microbiota were evaluated using a cyclophosphamide (Cy)-induced mice model. It was found that both CEPSs and EPSs-2 improved the body weight loss, immune organ indexes as well as the levels of TNF-α, IL-1β, IFN-γ and IgA, exhibiting potent immunoregulatory activity. Moreover, CEPSs and EPSs-2 not only attenuated the intestinal tissue damage, but also promoted the production of short-chain fatty acids and modulated the microbial composition by increasing the growth of Muribaculaceae, Prevotellaceae_UCG-001, Bacteroides, Parabacteroides and Tidjanibacter, while decreasing the relative abundances of Helicobacter, Bilophila, Mucispirillum, Lachnospiraceae, Ruminococcaceae and Clostridiales. These results indicated that the EPSs, especially EPSs-2, exhibited immunomodulatory activity associated with the modulation of gut microbiota to maintain gut homeostasis, which provided evidence for the development of novel potential prebiotics and immunomodulators.

The development of hardpan caps has great potential in rehabilitating sulfidic and metallic tailings, which may be accelerated by using exogenous Acidithiobacillus species. The present study aims to establish a bioaugmentation process with exogenous Acidithiobacillus species for accelerating the weathering of sulfidic minerals and formation of secondary mineral gels as precursors for hardpan structure development in a microcosm experiment. Exogenous Acidithiobacillus thiooxidans (ATCC 19377) and A. ferrooxidans (DSM 14882) were inoculated in a sulfidic Pb-Zn tailing containing negligible indigenous Acidithiobacillus species for accelerating the weathering of pyrite and metal sulfides. Microspectroscopic analysis revealed that the weathering of pyrite and biotite-like minerals was rapidly accelerated by exogenous Acidithiobacillus species, leading to the formation of secondary jarosite-like mineral gels and cemented profile in the tailings. Meanwhile, approximately 28% Zn liberated from Zn-rich minerals undergoing weathering was observed to be re-immobilized by Fe-rich secondary minerals such as jarosite-like mineral. Moreover, Pb-bearing minerals mostly remained undissolved, but approximately 30% Pb was immobilized by secondary Fe-rich minerals. The present findings revealed the critical role of exogenous Acidithiobacillus species in accelerating the precursory process of mineral weathering and secondary mineral formation for hardpan structure development in sulfidic Pb-Zn tailings.

Designing efficient single-atom catalysts (SACs) for oxygen evolution reaction (OER) is critical for water-splitting. However, the self-reconstruction of isolated active sites during OER not only influences the catalytic activity, but also limits the understanding of structure-property relationships. Here, we utilize a self-reconstruction strategy to prepare a SAC with isolated iridium anchored on oxyhydroxides, which exhibits high catalytic OER performance with low overpotential and small Tafel slope, superior to the IrO2. Operando X-ray absorption spectroscopy studies in combination with theory calculations indicate that the isolated iridium sites undergo a deprotonation process to form the multiple active sites during OER, promoting the O–O coupling. The isolated iridium sites are revealed to remain dispersed due to the support effect during OER. This work not only affords the rational design strategy of OER SACs at the atomic scale, but also provides the fundamental insights of the operando OER mechanism for highly active OER SACs., Direct observation of the atomic and electronic structure of a single-atom catalyst is essential. Here, the authors report an oxyhydroxide stabilized iridium catalyst with superior oxygen evolution catalytic activity and identify the isolated iridium sites which promote the H2O attack and O–O coupling.

Crude polysaccharides from Coralline pilulifera (CPPS) were purified by DEAE Sepharose Fast Flow to afford four purified fractions of CPPS-1, CPPS-2, CPPS-3 and CPPS-4. The physicochemical property and digestion under simulated saliva, gastric, small intestinal conditions and fermentation of CPPS-3 were investigated in vitro. CPPS-3 was composed of glucose and galactose, its molecular weight was 716 kDa, the contents of carbohydrate, protein, uronic acid and sulfuric radical were 67.84%, 1.00%, 0.38% and 9.07%, respectively. There was no significant difference on the molecular weight and content of reducing sugars, and no monosaccharides were released after saliva, gastric and small intestine digestion, indicating that CPPS-3 could not be digested under the simulated digestion conditions. After fermentation for 24 h in vitro, the carbohydrate content was decreased to 30.76%, suggesting that CPPS-3 could be broken down and utilized by gut microbiota. Some health-promoting gut microbiota such as Bacteroides, Megamonas and Veillonella were significantly enhanced, suggesting that CPPS-3 could regulate the composition of gut microbiota. What's more, CPPS-3 could promote the productions of short-chain fatty acids (22.17 and 16.17 mmol/L at 24 h for CPPS-3 and control, respectively), which could contribute to host health. Therefore, CPPS-3 is expected to be developed as functional foods.

Circular RNAs (circRNAs) have been found to be important mediators of many biological processes in the growth and metastasis of various cancers. However, the potential roles of most circRNAs in the progression of bladder cancer remain unclear. In this research, we investigate the role of circKIF4A (hsa_circ_0007255) in the development and progression of bladder cancer. Detected by qRT-PCR analysis, circKIF4A was significantly upregulated in bladder cancer tissues and cell lines. We conducted CCK-8, colony-formation, transwell and mouse xenograft assays to explore the function of circKIF4A in bladder cancer. Functionally, knockdown of circKIF4A inhibited the proliferation and colony-formation ability of bladder cancer cells. Migration and metastatic ability were dramatically decreased after transfection with small interfering RNA targeting circKIF4A in both in vitro and in vivo assays. Mechanically, luciferase reporter assays and RNA immunoprecipitation assays were carried out to elucidate the underlying molecular mechanism of circKIF4A. The results revealed that circKIF4A sponges miR-375/1231 to promote bladder cancer progression by upregulating NOTCH2. Generally, our research unveils the essential role of circKIF4A-miR-375/1231-NOTCH2 axis in bladder cancer progression possibly via the competing endogenous RNA mechanism.

Organo-mineral association and water-stable aggregation in finely textured tailings are critically important to the eco-engineered soil formation from alkaline Fe ore tailings for sustainable mine site rehabilitation. Arbuscular mycorrhizal (AM) symbiosis plays important roles in soil aggregate formation and organic matter (OM) stabilization. However, it is unknown if AM symbiosis could enhance aggregate formation and OM stabilization in alkaline Fe ore tailings. The present study aimed to investigate the establishment of AM symbiosis and their role in tailing aggregate formation coupled with OM stabilization, as well as the underlying mechanisms. After initial eco-engineering (OM amendment and pioneer plant cultivation) to improve physicochemical conditions for plant survival, Sorghum spp. Hybrid cv. Silk inoculated with/without AM fungi (Glomus spp.) were cultivated in the tailings under glasshouse conditions for 14 weeks. The results indicated that AM fungi formed symbiotic association with Sorghum spp. plants, improved mineral nutrient (e.g., P) acquisition and root growth in the eco-engineered tailings. The AM symbiosis significantly improved aggregate formation. The association of organic carbon and nitrogen with tailing minerals of the aggregates was enhanced by the AM symbiosis. As revealed by synchrotron-based C 1 s near edge X-ray absorption fine structure (C 1 s NEXAFS) and Fe K edge X-ray absorption fine structure (Fe K edge XAFS) spectroscopy, the AM symbiosis favoured carboxyl and aromatic C association with secondary Fe-Si minerals, which may have been formed from AM driven mineral weathering. Overall, the study revealed that the AM symbiosis could not only improve the growth of pioneer plant species in the early eco-engineered tailings, but also advance soil formation through enhancing organic C and N sequestration and physical structure development via water-stable aggregation. These findings help to advance our understanding of the importance of AM symbiosis in the eco-engineering of tailings into functional soil (or technosols) for sustainable rehabilitation of Fe-ore tailings.

Single-atom catalysts (SACs) offer us new opportunities for capturing the reaction intermediates, identifying the active sites, and even monitoring the structural and electronic evolution of catalytic centers. However, the lack of model SACs and atomic-resolution operando spectroscopic techniques greatly limits our comprehension of the nature of catalysis. Herein, based on the newly designed model single-Fe-atom catalyst with well-controlled site density and coordination environment, we have explored the exact structure of catalytic centers and provided insights into a novel spin-crossover-involved mechanism for oxygen reduction reaction (ORR) using operando Raman, X-ray absorption spectroscopies, as well as the newly developed operando 57Fe Mossbauer technique. Combining with theoretical studies, the single-Fe-atom site of N-FeN4C10 moiety, with preferential reaction to O2 near the onset potential, is shown as a more active site for ORR. Moreover, the potential-relevant dynamic cycles of both the geometric structure and electronic configuration of reactive single-Fe-atom moieties are evidenced via directly capturing the peroxido (*O2-) and hydroxyl (*OH-) intermediates under in-situ ORR conditions. Our results provide proof-of-concept for the integration of operando techniques and SACs, which directs the way towards the electronic-level insight into catalytic centers and underlying reaction mechanism.

The anamorphic taxon Anacacumisporium appendiculatum gen. et sp. nov. is described and illustrated from fallen branches of decaying wood in Hainan, China. Anacacumisporium is characterized by pigmented, transversely septate, appendaged conidia and conidiophores that are brown, macronematous, mononematous and that bear one (or more ) integrated phialide at the tip. Complete sequences of internal transcribed spacer (ITS) and partial sequences of nuc 28S rDNA (28S) genes are provided. The new fungus is compared with morphologically similar genera of hyphomycetes. Phylogenetic analyses revealed its placement in the Chaetosphaeriaceae. A key to the novel genus and morphologically similar genera is given.

Lycium barbarum has been considered as a valuable old herbal medicine that has been widely used in Asian country. Lycium barbarum polysaccharides (LBPs), a major component, are proved to possess a variety of beneficial effects including neuro-protection, anticancer properties and anti-aging as well as immune modulation. However, the biological functions and inductive mechanisms of LBP towards the mesenchymal stem cell (MSC) differentiation process have not been fully investigated. In this study, we found that the expression of specific marker of neuron cells including β-tubulin III, NF200 and synapsin was up-regulated when cultured with LBP after 7 days during differentiation of bone marrow-derived MSCs to neuron-like cells, while the pluripotency marker such as Oct4 and Klf4 were down-regulated. By using the staining of phalloidine, the cytoskeleton of the stem cells induced by LBP exhibited lengthening character, resulting in the induction effect of LBP towards BMSCs. Overall, we demonstrate that the directional differentiation process of BMSCs can be partially regulated by LBP. We conclude that this strategy may provide opportunities for physician to improve the effectiveness of stem cell therapy for the treatment of degenerative cerebrovascular disease.

RhoA has been implicated in diverse cellular functions and is a potential cancer therapeutic target. Through virtual screening, we have identified a RhoA inhibitor, DDO-5701. DDO-5701 has an affinity to RhoA at the micromolar level in vitro. By structural modifications, considering the binding activity and synthesis ease of DDO-5701, 17 compounds were designed and synthesized accordingly. Among these compounds, 4 compounds (DDO-5713, DDO-5714, DDO-5715, DDO-5716) exhibited higher RhoA inhibition activities than DDO-5701, while DDO-5716 can effectively reverse the functions of breast cancer cells regulated by RhoA. Thus, the rationally designed small molecule inhibitor of RhoA (DDO-5716) is useful for studying the physiological and pathological roles of Rho GTPase. However, DDO-5701 is an approved drug – proglumide, which makes it and its derived compound DDO-5716 more likely to be well tolerated in humans and could quickly lead to further clinical development.

p53 protein is a prominent tumor suppressor to induce cell cycle arrest, apoptosis and senescence, which attracts significant interest to cancer treatment. Therefore, it would be particularly important to restore the wild-type p53 that retains latent functions in the approximately 50% of tumors. MDM2 (murine double minute 2), the principal cellular antagonist of p53, has long been believed to suppress p53 activity through two main mechanisms: promoting degradation via its E3 ligase activity and masking p53 transcriptional activation by direct binding. Targeting MDM2 E3 ligase activity is becoming a potential antitumor strategy resulting from MDM2's decisive role in controlling the fate of p53: p53 is going to degradation when entrapped into MDM2-mediated ubiquitination, where p53 can escape by abrogating MDM2 E3 ligase activity using regulators. The intensive focus on regulating MDM2 ubiquitin E3 ligase activity has led to the rapid progress of its inhibitors, which may be possible to help p53 escape from degradation and restore its function to control tumor growth. This review summarizes the current inhibitors of MDM2 E3 ligase in cancer therapy based on the understanding the regulation of MDM2 E3 ubiquitin ligase activity, including post-translational modification, interactions between MDM2 and its cofactors, and regulation of MDM2 stability.

The DNA-binding one zinc finger (Dof) family is a classic plant-specific zinc-finger transcription factor family, which is involved in many important processes, including seed maturation and germination, plant growth and development, and light responses. Investigation of the Medicago truncatula genome revealed 42 putative Dof genes, each of which holds one Dof domain. These genes were classified into four groups based on phylogenetic analysis, which are similar to the groups reported for Arabidopsis and rice. Based on genome duplication analysis, it was found that the MtDof genes were distributed on all chromosomes and had expanded through tandem gene duplication and segmental duplication events. Two main duplication regions were identified, one from tandem duplication and another from segmental duplication. By analyzing high-throughput sequencing data from M. truncatula, we found that most of the MtDof genes showed specific expression patterns in different tissues. According to cis-regulatory element analysis, these MtDof genes are regulated by different cis-acting motifs, which are important for the functional divergence of the MtDof genes in different processes. Thus, using genome-wide identification, evolution, and expression pattern analysis of the Dof genes in M. truncatula, our study provides valuable information for understanding the potential function of the Dof genes in regulating the growth and development of M. truncatula.

During our continuous survey (2012–2016) of saprobic hyphomycetes from dead branches in the forest ecosystems of southern China, we collected several acrodictys-like species. Acrodictys-like species are characterized by darkly pigmented and muriform conidia produced from holoblastic conidiogenous cells on macronematous, mononematous, cylindrical and unbranched or infrequently branched conidiophores. Phylogenetic analyses of ncLSU, ncSSU, ITS and tub2 sequence data lead us to propose two novel families in Sordariomycetes, Acrodictyaceae and Junewangiaceae. In addition, a new species, Acrodictys hainanensis, two new combinations, Junewangia queenslandica and Distoseptispora martinii, three new Chinese records, Acrodictys liputii, A. peruamazonensis and Junewangia sphaerospora are introduced. Two names, Acrodictys globulosa and A. malabarica, are resurrected.

EHD2, a member of the Eps15 homology domain (EH domain) family, is important for protein interactions during vesicular trafficking. Previous studies have proved that EHD2 can regulate trafficking from the plasma membrane in the process of endocytosis. However, its function in central nervous system diseases is still with limited understanding. In this frame, we found that EHD2 expression was upregulated in the perihematomal caudate in adult rats after intracerebral hemorrhage (ICH). Double immunofluorescence staining revealed that EHD2 was colocalized with neurons and activated microglias after ICH. Besides, we detected that neuronal apoptosis markers (TUNEL and caspase-3), and microglial activation marker (CD68), also known as a marker of macrophage, were colocated with EHD2. The vitro study also indicated that EHD2 was linked with neuronal apoptosis and microglial phagocytosis. All our findings suggested that EHD2 might be involved in the pathophysiology of ICH.

Fuzhuan brick tea is characterized in unique “golden flower” of Aspergillus cristatus. However, little information is available for intra- and extra-cellular polysaccharides from A. cristatus (IPS and EPS, respectively). Therefore, simulated digestion and fermentation in vitro were carried out to investigate whether IPS and EPS could be broken down in digestive system. Furthermore, their interactions with gut microbiota were investigated. It was found that IPS and EPS were utilized by gut microbiota to metabolize into short-chain fatty acids (SCFAs). The contents of total acids (SCFAs and lactic acid) in IPS and EPS groups increased to 39.30 ± 0.66 and 45.18 ± 1.68 mmol/L, respectively. IPS and EPS could obviously modulate the composition of gut microbiota. Thereinto, IPS could maintain the growth of OTU26 and OTU69 (Prevotella) and increase the relative levels of OTU30, OTU31, OTU35 and OTU56 (Bacteroides), whereas, EPS could maintain the growth of OTU26 and OTU48 (Prevotella) and increase the relative levels of OTU35, OTU56 and OTU63 (Bacteroides). OTU26, OTU61 and OTU69 (Prevotella) showed significantly positive correlations with propionic acid, OTU48 (Prevotella) and OTU56 (Bacteroides) showed significantly positive correlations with acetic acid. Hence, IPS and EPS could regulate the composition of gut microbiota, thereby promote production of SCFAs.

Corynesporopsis curvularioides sp. nov. is described and illustrated from specimens collected on dead branches in Guangxi Province. The fungus is characterized by its terminal, monotretic conidiogenous cells with catenate, obclavate to ellipsoidal, pale brown to brown, 15-35 × 7-11 μm, 1-5-euseptate conidia. Brachysporiellina pulneyensis, Catenularia cubensis and Kylindria excentrica are newly recorded from China. Biodiversity / conidial fungi / hyphomycetes / systematics / taxonomy

Wild type p53-induced phosphatase 1 (Wip1) is a phosphatase which belongs to protein phosphatase type 2C family, which have been predominantly linked to cell growth and to cellular stress signaling. Numerous downstream targets of Wip1 have been identified, and genetic studies confirm that some play a part in tumorigenesis. Recent evidence highlights a new role for Wip1 in the regulation of NF-κB p65, which indicated that it might play a critical role in immune system. However, its regulation role in central nervous system (CNS) remains poorly understood. To elaborate whether Wip1 was involved in CNS injury, we performed a neuroinflammatory model by lipopolysaccharide (LPS) lateral–ventral injection in adult rats. Wip1 expression was strongly upregulated in active astrocytes in inflamed brain cortex. In vitro studies indicated that the upregulation of Wip1 may be involved in the subsequent astrocytic activation following LPS exposure, and knockdown of Wip1 in primary astrocytes by siRNA showed that Wip1 inhibited the synthesis of TNF-α. Collectively, these results suggested that Wip1 may be important in host defense in CNS immune response, which might provide a potent therapeutic target of neuroinflammation.

The members of high-temperature requirement (HtrA) family are evolutionarily conserved serine proteases that combine a trypsin-like protease domain with at least one PDZ interaction domain. HtrA2, a special one, is mainly located in mitochondria and required for maintaining homeostasis. Once released into cytoplasm, HtrA2 contributes to apoptosis via caspase-dependent and -independent pathways. Accumulating evidence has showed its pro-apoptotic effect in cancers and central nervous system (CNS) diseases. However, the distribution and function of HtrA2 in CNS diseases remains to be further explored. To investigate HtrA2's roles in the pathophysiology of intracerebral hemorrhage (ICH), an ICH rat model was established and assessed by behavioral tests. Western blot and immunohistochemistry revealed a remarkable up-regulation of HtrA2 surrounding the hematoma after ICH; and immunofluorescence showed HtrA2 was strikingly increased in neurons, but not in astrocytes and oligodendrocytes. Terminal deoxynucleotidyl transferase-mediated biotinylated-dUTP nick-end labeling staining suggested the involvement of HtrA2 in neuronal apoptosis after ICH. Additionally, HtrA2 co-localized with active-caspase-3 around the hematoma and the expression of active-caspase-3 was parallel with that of HtrA2 in a time-dependent manner. Furthermore, hemin was used to stimulus a neuronal cell line PC12 to mimic ICH model in vitro. We analyzed the relationship of HtrA2 with X-linked inhibitor of apoptosis protein (XIAP) in PC12 cells by Western blot, immunofluorescence and co-immunoprecipitation. The connection of HtrA2 with XIAP was strengthened in apoptotic cells after hemin treatment. Thus, we speculated that HtrA2 might exert an important function in regulating caspase-dependent neuronal apoptosis through interacting with XIAP following ICH.

αB-crystallin (αBC) is involved in diverse cellular activities. Previous studies demonstrated that αBC had anti-apoptotic and proliferation-promoting effects in multiple diseases. Here, we explored the αBC's roles in the pathophysiology of intracerebral hemorrhage (ICH). An ICH rat model was established and assessed by behavioral tests. Using Western blot and immunohistochemistry, significant up-regulation of αBC was found in neurons and astrocytes in brain areas surrounding the hematoma following ICH. Increase of αBC expression was found to be accompanied by the increased expression of proliferating cell nuclear antigen (PCNA), p53, Bax, and active-caspase-3. αBC was co-localized with PCNA in astrocytes or active-caspase-3 in neurons, suggesting its role in astrocyte proliferation and neuronal apoptosis. Our in vitro study, using αBC RNA interference in PC12 cells, indicated that αBC might exert its anti-apoptotic function in neuronal apoptosis. Thus, αBC may play a role in protecting the brain from secondary damage following ICH.

Saline-alkaline stress, caused by high levels of harmful carbonate salts and high soil pH, is a major abiotic stress that affects crop productivity. Alfalfa is a widely cultivated perennial forage legume with some tolerance to biotic and abiotic stresses, especially to saline-alkaline stress. To elucidate the mechanism underlying plant saline-alkaline tolerance, we conducted transcriptome analysis of whole alfalfa seedlings treated with saline-alkaline solutions for 0 day (control), 1 day (short-term treatment), and 7 days (long-term treatment) using ion torrent sequencing technology. A transcriptome database dataset of 53,853 unigenes was generated, and 2,286 and 2,233 genes were differentially expressed in the short-term and long-term treatment, respectively. Gene ontology analysis revealed 14 highly enriched pathways and demonstrated the differential response of metabolic pathways between the short-term and long-term treatment. The expression levels of 109 and 96 transcription factors were significantly altered significantly after 1 day and 7 days of treatment, respectively. Specific responses of peroxidase, flavonoids, and the light pathway component indicated that the antioxidant capacity was one of the central mechanisms of saline-alkaline stress tolerance response in alfalfa. Among the 18 differentially expressed genes examined by real time PCR, the expression levels of eight genes, including inositol transporter, DNA binding protein, raffinose synthase, ferritin, aldo/keto reductase, glutathione S-transferase, xyloglucan endotrans glucosylase, and a NAC transcription factor, exhibited different patterns in response to saline and alkaline stress. The expression levels of the NAC transcription factor and glutathione S-transferase were altered significantly under saline stress and saline-alkaline stress; they were upregulated under saline-alkaline stress and downregulated under salt stress. Physiology assays showed an increased concentration of reactive oxygen species and malondialdehyde and a decreased content of chlorophyll, indicating that anti-oxidation and detoxification play an important role in response to saline-alkaline stress. Overall, the transcriptome analysis provided novel insights into the saline-alkaline stress tolerance response mechanisms in alfalfa.

Vascular endothelial growth factor-C (VEGF-C) has been found to be significantly associated with lymphangiogenesis and regional lymph node metastasis in various human tumors. The present work was aimed to explore the role of VEGF-C in malignant progression of human bladder cancer T24 cell line. First, the expression of VEGF-C in T24 cells was detected by western blotting. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was employed to measure the cellular proliferation after treatment with various concentrations of recombinant human VEGF-C (rhVEGF-C). Then, lentivirus vector-based RNA interference (RNAi) was used to inhibit VEGF-C expression of T24 cells. The alterations of T24 cells regarding proliferation, invasiveness, and the apoptosis induced by mitomycin C (MMC) were evaluated. The results showed that the proliferation rate of T24 cells rose from 27.3% to 65.0%, with increasing rhVEGF-C concentration. T24 cells stably transfected with VEGF-C small interference RNA showed 85% reduction in VEGF-C mRNA expression (p 0.05). The VEGF-C protein level was significantly downregulated (p 0.05) and the growth and invasiveness were also inhibited (p 0.05) compared with the control group. Further, the inhibition of VEGF-C expression markedly enhanced the apoptosis of T24 cells induced by MMC (p 0.05). These were associated with the decreased ratio of Bcl-2/Bax, activation of Caspase-3, decreased expression of MMP-9, as well as the downregulation of phosphorylated p38 MAPK and Akt. The present study suggests that VEGF-C can enhance the proliferation and invasiveness of bladder cancer T24 cells, which is due to suppression of apoptosis and facilitation of migration, accompanied with upregulation of p38 MAPK and Akt phosphorylation. RNAi targeting VEGF-C could effectively suppress malignant progression and enhance chemosensitivity of T24 cells. Thus, inhibition of VEGF-C expression is a potential and promising therapeutic strategy for bladder cancer.

p53-independent malignant cancer is still severe health problem of human beings. HIF-1 pathway is believed to play an important role in the survival and developing progress of such cancers. In the present study, with the aim to inhibit the proliferation of p53-independent malignant cells, we disclose the optimization of 6a, the starting compound which is discovered in the screening of in-house compound collection. The structure-activity relationship (SAR) is summarized. The most potent derivative 8d, inhibits the proliferation of both p53-null and p53-mutated cells through inhibition of HIF-1 pathway. Our findings here provide a new chemotype in designing potent anticancer agent especially against those p53-independent malignant tumors.

// Chao Zhang 1, 2 , Hui-Jie Wang 1, 2 , Qi-Chao Bao 1, 2 , Lei Wang 1, 2 , Tian-Kun Guo 1, 2 , Wei-Lin Chen 1, 2 , Li-Li Xu 1, 2 , Hai-Shan Zhou 1, 2 , Jin-Lei Bian 1, 2 , Ying-Rui Yang 1, 2 , Hao-Peng Sun 1, 2, 3 , Xiao-Li Xu 1, 2, 3 , Qi-Dong You 1, 2, 3 1 Jiangsu Key Laboratory of Drug Design and Optimization, China Pharmaceutical University, Nanjing, 210009, China 2 State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, 210009, China 3 Department of Medicinal Chemistry, School of Pharmacy, China Pharmaceutical University, Nanjing, 210009, China Correspondence to: Xiao-Li Xu, email: xuxiao_li@126.com Qi-Dong You, email: youqidong@gmail.com Keywords: NRF2, breast cancer, RhoA/ROCK pathway, cell proliferation, metastasis Received: January 04, 2016 Accepted: September 26, 2016 Published: October 04, 2016 ABSTRACT Nuclear factor erythroid 2-related factor (NRF2) is an important transcription factor in oxidative stress regulation. Overexpression of NRF2 is associated with human breast carcinogenesis, and increased NRF2 mRNA levels predict poor patient outcome for breast cancer. However, the mechanisms linking gain of NRF2 expression and poor prognosis in breast cancer are still unclear. Here, we provide evidence that NRF2 deletion inhibits proliferation and metastasis of breast cancer cells by down-regulating RhoA. Restoration of RhoA in MCF7 and MDA-MB-231 cells induced NRF2 knockdown-suppressed cell growth and metastasis in vitro, and NRF2 silencing suppressed stress fiber and focal adhesion formation leading to decreased cell migration and invasion. Mechanistic studies showed that NRF2 binds to the promoter region of estrogen-related receptor α (ERR1) and may function as a silencer. This may enhance RhoA protein stability and lead to RhoA overexpression in breast cancer cell. Our findings indicate that NRF2 silencing-mediated reduction of RhoA expression contributes, at least in part, to the poor outcome of breast cancer patients with high NRF2 expression.

Two new quaternary isoquinoline alkaloids, saxicolalines A (1) and N-methylnarceimicine (2), together with sixteen known alkaloids (3 - 18) were isolated from Corydalis saxicola Bunting. The structures of saxicolalines A (1) and N-methylnarceimicine (2) were established based on spectral methods including 1D, 2D NMR (COSY, HMQC, HMBC) and HR-ESI-MS. The anti-HBV activities of ten main alkaloids were evaluated. Among the tested compounds, dihydrochelerythrine (8) exhibited the most potent activity against HBsAg and HBeAg secretions with IC500.05 microM, SI3.5, respectively.

Hypoxia-inducible factor-1 (HIF-1), a heterodimeric (containing α and β subunits) transcription factor, is involved in hypoxia response pathway that regulates the expression of many tumorrelated genes. The stabilized HIF-1 heterodimer couples to the general co-activators p300/CBP (CREB binding protein), forming an active transcription factor to initiate hypoxic responses. Inhibiting the transcription factor-coactivator HIF-1α-p300/CBP interaction represents an attractive approach for blocking hypoxia pathway in tumors. Recently, diverse HIF-1α-p300/CBP inhibitors have been designed and their anti-tumor activities have been evaluated. The developments of inhibitors of HIF-1α- p300/CBP are discussed in this review. An outline of structures and biological activities of these inhibitors can be traced, along with the approaches for inhibitors discovery. The challenges in identifying novel and selective potent inhibitors of HIF-1α-p300/CBP are also put forward.

Atrokylindriopsis setulosa gen. et sp. nov. produces pigmented, septate, setulate conidia that are attached to the monophialidic conidiogenous cells of unbranched, brown, macronematous conidiophores at the midpoint of their long side during conidiogenesis, giving the appearance of a ‘T’. Internal transcribed spacer region (ITS) and partial 28S rDNA sequences indicate placement of Atrokylindriopsis in the Chaetothyriales.

The identification of the H3K4 trimethylase, PRDM9, as the gene responsible for recombination hotspot localization has provided considerable insight into the mechanisms by which recombination is initiated in mammals. However, uniquely amongst mammals, canids appear to lack a functional version of PRDM9 and may therefore provide a model for understanding recombination that occurs in the absence of PRDM9, and thus how PRDM9 functions to shape the recombination landscape. We have constructed a fine-scale genetic map from patterns of linkage disequilibrium assessed using high-throughput sequence data from 51 free-ranging dogs, Canis lupus familiaris. While broad-scale properties of recombination appear similar to other mammalian species, our fine-scale estimates indicate that canine highly elevated recombination rates are observed in the vicinity of CpG rich regions including gene promoter regions, but show little association with H3K4 trimethylation marks identified in spermatocytes. By comparison to genomic data from the Andean fox, Lycalopex culpaeus, we show that biased gene conversion is a plausible mechanism by which the high CpG content of the dog genome could have occurred., Updated version, with significant revisions

Nuclear factor of activated T-cells, cytoplasmic 4 (NFATc4), a transcriptional factor, is involved in the control about the flow of genetic information and the modulation of diverse cellular activities. Accumulating evidence has demonstrated that NFATc4 exerted a pro-apoptotic effect in multiple diseases. Here, we explored the NFATc4’s roles during the pathophysiological processes of intracerebral hemorrhage (ICH). An ICH rat model was built and evaluated according to behavioral testing. Using Western blot, immunohistochemistry, and immunofluorescence, significant up-regulation of NFATc4 was found in neurons in brain areas surrounding the hematoma following ICH. Increasing NFATc4 expression was found to be accompanied by the up-regulation of Fas ligand (FasL), active caspase-8, and active caspase-3, respectively. Besides, NFATc4 co-localized with active caspase-3 in neurons, indicating its role in neuronal apoptosis. Our in vitro study, using NFATc4 RNA interference in PC12 cells, further confirmed that NFATc4 might exert its pro-apoptotic function in neuronal apoptosis through extrinsic pathway. Thus, NFATc4 may play a role in promoting the brain secondary damage following ICH.

Using a newly developed strategy whose key step is the regioselective propargylation of hydroxyxanthone substrates, 99 structurally diverse Garcinia natural-product-like xanthones based on gambogic acid were designed and synthesized and their in vitro antitumor activity was evaluated. A set of 40 related compounds was chosen for determination of their physicochemical properties including polar surface area, log D7.4, aqueous solubility, and permeability at pH 7.4. In the light of the in vitro antitumor activity and the physicochemical properties, two compounds were advanced into in vivo efficacy experiments. The antitumor activity of compound 112, administered po, showed more potent in vivo oral antitumor activity than gambogic acid.

CBLL1 (Casitas B-lineage lymphoma-transforming sequence-like protein 1) also known as Hakai, was originally identified as an E3 ubiquitin-ligase for the E-cadherin complex. Recent data have provided evidences for novel biological functional role of CBLL1 during tumor progression and other diseases. However, its distribution and function in the central nervous system (CNS) remains unclear. In this study, we found CBLL1 was significant up-regulation in cerebral cortex after LPS administration and immunofluorescent labeling indicated that CBLL1 was localized striking in the neurons. We also investigated co-staining of CBLL1 and active-caspase-3 and cyclin D1 in the cerebral cortex following LPS administration. Based on our data, we speculated that CBLL1 might play an important role in neuronal apoptosis following LPS administration and might provide a basis for the further study on its role in cell cycle re-entry in neuroinflammation in CNS.

Ras homolog enriched in the brain (Rheb) is a homolog of Ras GTPase that regulates cell growth, proliferation, and cell cycle via mammalian target of rapamycin (mTOR). Recently, it has been confirmed that Rheb activation not only promotes cellular proliferation and differentiation but also enhances cellular apoptosis in response to diverse toxic stimuli. However, the function of Rheb in the central nervous system (CNS) is still with limited understanding. To elaborate whether Rheb was involved in CNS injury, we performed a neuroinflammatory model by lipopolysaccharide (LPS) lateral ventral injection in adult rats. Upregulation of Rheb was observed in the brain cortex by performing western blotting and immunohistochemistry. Double immunofluorescent staining demonstrated that Rheb was mainly in active astrocytes and neurons. PCNA and active caspase-3 were upregulated, and co-labeling with Rheb, which indicated that Rheb might be relevant to astrocytic proliferation and neuronal apoptosis following the inflammatory response by LPS-induced. Furthermore, we also found that the expression profiles of cyclinD1 and CDK4 were parallel with that of Rheb in a time-space dependent manner. Finally, knocking down Rheb by siRNA and treatment with rapamycin or lovastatin showed that not only astrocytic proliferation decreased but also neuronal protection. Based on our data, we suggested that Rheb might play an important role in physiological and pathological functions following neuroinflammation caused by LPS, which might provide a potential target to the treatment of neuroinflammation.

To examine the expression of Egr-1, c-fos and cyclin D1 at both transcript and protein levels in esophageal carcinoma and to correlate the level of their expressions with precancerous and paracancerous esophageal lesions and esophageal carcinoma.In situ hybridization and immunohistochemistry were used respectively to detect the expression of mRNA and proteins of Egr-1, c-fos and cyclin D1 in 70 cases of esophageal squamous cell carcinoma and their corresponding para-cancerous mucosa and upper cut edge mucosa.In situ hybridization and immunohistochemistry showed positive staining of all three mRNAs in the cytoplasm and those of the proteins in nuclei. Overexpression of Egr-1, c-fos and cyclin D1 mRNAs and their proteins was found in dysplasia and squamous carcinomas. The expression level of Egr-1 and c-fos was high, and cyclin D1 was low in dysplasia mucosa, whereas the expression of Egr-1 was decreased, c-fos was maintained and cyclin D1 was increased in the cancers. The expression of both c-fos and cyclinD1 was consistent between the mRNA and protein in their corresponding high expression lesions.The expression of Egr-1, c-fos and cyclin D1 varies in esophageal precancerous lesions and cancer tissues, suggesting an involvement of these genes in the development of esophageal carcinoma.