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The vertebrate cardiovascular system is made up by a hierarchically structured network of highly specialised blood vessels. This network emerges during early embryogenesis and evolves in size and complexity concomitant with embryonic growth and organ formation. Underlying this plasticity are actin-driven endothelial cell behaviours, which allow endothelial cells to change their shape and move within the vascular network. In this review, we discuss the cellular and molecular mechanisms involved in vascular network formation and how these intrinsic mechanisms are influenced by haemodynamic forces provided by pressurized blood flow. While most of this review focusses on in vivo evidence from zebrafish embryos, we also mention complementary findings obtained in other experimental systems.

The woolly flying squirrel, Eupetaurus cinereus, is among the rarest and least studied mammals in the world. For much of the 20th century it was thought to be extinct, until it was rediscovered in 1994 in northern Pakistan. This study outlines the first taxonomic and biogeographical review of the genus Eupetaurus, which until now has contained only a single species. Careful review of museum specimens and published records of Eupetaurus demonstrates that the genus occurs in three widely disjunct areas situated on the western (northern Pakistan and north-western India), north-central (south-central Tibet, northern Sikkim and western Bhutan) and south-eastern margins (north-western Yunnan, China) of the Himalayas. Taxonomic differentiation between these apparently allopatric populations of Eupetaurus was assessed with an integrative approach involving both morphological examinations and molecular phylogenetic analyses. Phylogenetic reconstruction was implemented using sequences of three mitochondrial [cytochrome b (Cytb), mitochondrially encoded 12S and 16S ribosomal RNA (12S, 16S)] and one nuclear [interphotoreceptor retinoid-binding protein (IRBP)] gene fragment. Morphological assessments involved qualitative examinations of features preserved on museum skins and skulls, supplemented with principal components analysis of craniometric data. Based on genetic and morphological comparisons, we suggest that the three widely disjunct populations of Eupetaurus are each sufficiently differentiated genetically and morphologically to be recognized as distinct species, two of which are described here as new.

Multiple myeloma (MM) is a plasma cell malignancy of bone marrow. In the present study, we aimed to study the function and potential mechanism of the antisense non-coding RNA in the INK4 Locus (ANRIL) in MM. The expression levels of ANRIL in MM patients and healthy donors were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). The effects and mechanisms of ANRIL in MM were evaluated by cell viability assay, BrdU incorporation assay, tumor xenograft model, flow cytometry, western blot, RNA immunoprecipitation (RIP), transcriptome RNA sequencing, and chromatin immunoprecipitation (ChIP). We found that ANRIL was upregulated in MM patients and cell lines, and associated with advanced international staging system (ISS) stage and poor overall survival. Enforced ANRIL expression promoted proliferation and tumor xenograft growth of MM cells, while knockdown of ANRIL exhibited opposite effects. Moreover, ANRIL overexpression increased the half-maximal inhibitory concentration (IC50) of bortezomib and reduced bortezomib-induced apoptosis in MM cells. ANRIL was found to accumulate in the nuclei of MM cells, and interact with EZH2 by RIP assay. Transcriptome RNA sequencing identified PTEN as a target of ANRIL in MM cells. In the ChIP assay, knockdown of ANRIL reduced EZH2 occupancy and H3K27me3 binding to the promoter region of PTEN. Furthermore, EZH2 knockout or PTEN restoration abrogated the effects caused by ANRIL overexpression in MM cells. Our results indicated that ANRIL exerted oncogenic functions and conferred chemoresistance of MM cells by EZH2-mediated epigenetically silencing of PTEN.

Insecticidal crystal (Cry) proteins produced by genetically modified rice that enter the soil via pollen dispersal, plant residues, and root exudation may disturb soil health. In the present study, we assessed the influences of transgenic Bt rice (i.e., HH1 with Cry1Ab/Cry1Ac) cultivation on the dynamics of soil carbon and nutrients under field conditions during 2013-2016. Transgenic treatments (transgenic Bt rice vs. its parental line (i.e., MH63) of non-Bt rice) have no consistently significant effects on soil property, including available nitrogen, available phosphorus, available potassium, total nitrogen, and total phosphorus, while apparent seasonal changes were observed. Besides, the variations of soil nutrients in the paddy field of transgenic Bt rice did not exceed their resistance capacities, except total organic carbon (TOC; RS (resistance) = 1.51) and total potassium (TK; RS = 2.62) in 2013 and TK (RS = 1.94) in 2014. However, the TOC and soil nutrient of TK in the paddy field of transgenic Bt rice have recovered to the pre-perturbation status after harvest (RL (resilience) = 1.01, F = 0.01, P = 0.91; RL = 0.98, F = 0.34, P = 0.58; RL = 0.99, F = 1.26, P = 0.29). Moreover, the paddy yield of transgenic Bt rice was consistently higher than that of its parental line of non-Bt rice. These results suggested that the cultivation of transgenic Bt rice has no adverse impact on soil stability in terms of soil carbon and nutrients and paddy yield.

The evolution of increased competitive ability (EICA) hypothesis states that, when introduced in a novel habitat, invasive species may reallocate resources from costly quantitative defense mechanisms against enemies to dispersal and reproduction; meanwhile, the refinement of EICA suggests that concentrations of toxins used for qualitative defense against generalist herbivores may increase. Previous studies considered that only few genotypes were introduced to the new range, whereas most studies to test the EICA (or the refinement of EICA) hypotheses did not consider founder effects. In this study, genetic and phenotypic data of Chromolaena odorata populations sampled across native and introduced ranges were combined to investigate the role of postintroduction evolution in the successful invasion of C. odorata. Compared with native populations, the introduced populations exhibited lower levels of genetic diversity. Moreover, different founder effects events were interpreted as the main cause of the genetic structure observed in introduced ranges. Three Florida, two Trinidad, and two Puerto Rico populations may have been the sources of the invasive C. odorata in Asia. When in free of competition conditions, C. odorata plants from introduced ranges perform better than those from native ranges at high nutrient supply but not at low nutrient level. The differences in performance due to competition were significantly greater for C. odorata plants from the native range than those from the introduced range at both nutrient levels. Moreover, the differences in performance by competition were significantly greater for putative source populations than for invasive populations. Quantities of three types of secondary compounds in leaves of invasive C. odorata populations were significantly higher than those in putative source populations. These results provide more accurate evidence that the competitive ability of the introduced C. odorata is increased with postintroduction evolution.

Although the roles of long non-coding RNA (lncRNA) ANRIL (Antisense non-coding RNA in the INK4A locus) have been established in various tumors, its roles in mitochondrial metabolic reprogramming of hepatocellular carcinoma (HCC) cells are still unclear. This work aims to explore lncRNA ANRIL roles in regulating the mitochondrial metabolic reprogramming of liver cancer cells. First, we found that lncRAN ANRIL expression was significantly increased in HCC tissues or cells compared with the normal adjacent tissues and normal tissues or cells. Functional experiment showed that overexpression of lncRNA ANRIL promoted mitochondrial function in HCC cells, evident by the increased mitochondrial DNA copy numbers, ATP (Adenosine triphosphate) level, mitochondrial membrane potential, and the expression levels of mitochondrial markers, while ANRIL knockdown exerted the opposite effects. Mechanistically, lncRNA ANRIL acted as a competing endogenous RNA to increase ARL2 (ADP-ribosylationfactor-like 2) expression via sponging miR-199a-5p. Notably, the miR-199a-5p/ARL2 axis is necessary for ANRIL-mediated promoting effects on HCC cell mitochondrial function. This work reveals a novel ANRIL-miR-199a-5p-ARL2 axis in HCC cell progression, which might provide potential targets for HCC treatment.

Qing-Min Liu,1,* Yi-Yu He,2,* Li-Li Liu,3 Li-Kun Wang4 1Intensive Care Unit, Linyi People’s Hospital, Linyi, Shandong Province, People’s Republic of China; 2Department of Cardiovascular Disease, Renmin Hospital of Wuhan University, Wuhan, Hubei Province, People’s Republic of China; 3Department of Pathology, Linyi People’s Hospital, Linyi, Shandong Province, People’s Republic of China; 4Infection Control Center, Linyi People’s Hospital, Linyi, Shandong Province, People’s Republic of China*These authors contributed equally to this workCorrespondence: Li-Li LiuDepartment of Pathology, Linyi People’s Hospital, Linyi, Shandong Province, People’s Republic of ChinaEmail 13505495928@126.comLi-Kun WangInfection Control Center, Linyi People’s Hospital, Linyi, Shandong Province, People’s Republic of ChinaEmail Lkwang999@163.comIntroduction: Chronic hepatitis B (CHB) virus (HBV) infection has emerged as a global health burden affecting nearly 292 million people. Tenofovir alafenamide (TAF) is an effective treatment for CHB patients. However, the detailed mechanism underlying the antiviral activity of TAF remains unclear.Methods: In this study, we investigated the antiviral effect of exosomes derived from the serum of CHB patients treated with TAF (Exo-serum) and TAF-treated macrophages (MP) (Exo-MP(TAF)).Results: RNAseq analysis was also performed to determine the associated long non-coding RNAs (lncRNAs). The results demonstrated that both Exo-serum and Exo-MP(TAF) could be taken up by HepAD38 cells and exhibited potent antiviral activities, as manifested by significantly downregulating the levels of hepatitis B surface antigen, hepatitis B e antigen, HBV DNA, and covalently closed circular DNA. The antiviral effect of Exo-serum was more potent than those of TAF treatment alone. RNAseq analysis revealed that lncRNA HOTTIP was upregulated significantly in Exo-serum. Further, lncRNA HOTTIP knockdown reversed the antiviral effect of Exo-MP(TAF) on HepAD38 cells, whereas lncRNA HOTTIP knockdown exerted the opposite roles.Discussion: Taken together, these results suggest that exosomal lncRNA HOTTIP is essential for the antiviral activity of TAF and provide a novel understanding of the exosome-mediated mechanism underlying HBV infection.Keywords: chronic hepatitis B, tenofovir alafenamide, exosome, lncRNA HOTTIP

Nitrous oxide (N2 O) was previously deemed as a potent greenhouse gas but is actually an untapped energy source, which can accumulate during the microbial denitrification of nitric oxide (NO). Compared with the organic electron donor required in heterotrophic denitrification, elemental sulfur (S0 ) is a promising electron donor alternative due to its cheap cost and low biomass yield in sulfur-driven autotrophic denitrification. However, no effort has been made to test N2 O recovery from sulfur-driven denitrification of NO so far. Therefore, in this study, batch and continuous experiments were carried out to investigate the NO removal performance and N2 O recovery potential via sulfur-driven NO-based denitrification under various Fe(II)EDTA-NO concentrations. Efficient energy recovery was achieved, as up to 35.5%-40.9% of NO was converted to N2 O under various NO concentrations. N2 O recovery from Fe(II)EDTA-NO could be enhanced by the low bioavailability of sulfur and the acid environment caused by sulfur oxidation. The NO reductase (NOR) and N2 O reductase (N2 OR) were inhibited distinctively at relatively low NO levels, leading to efficient N2 O accumulation, but were suppressed irreversibly at NO level beyond 15 mM in continuous experiments. Such results indicated that the regulation of NO at a relatively low level would benefit the system stability and NO removal capacity during long-term system operation. The continuous operation of the sulfur-driven Fe(II)EDTA-NO-based denitrification reduced the overall microbial diversity but enriched several key microbial community. Thauera, Thermomonas, and Arenimonas that are able to carry out sulfur-driven autotrophic denitrification became the dominant organisms with their relative abundance increased from 25.8% to 68.3%, collectively.

Target identification is one of the most important bases for novel pesticide development; pyruvate kinase (PK) was discovered as a potent fungicide target in our previous studies. To continue the PK-based fungicide development, novel isothiazole-purine derivatives were rationally designed and synthesized. Bioassay results showed that compound 5ai displayed excellent in vitro activity against Rhizoctonia solani with an EC50 of 1.5 μg/mL, which was superior to those of positive controls diflumetorim with its EC50 of 19.8 μg/mL and PK-based lead YZK-C22 with its EC50 of 4.2 μg/mL. Compounds 3b (5.2 μg/mL) and 3c (4.5 μg/mL) displayed better activities against Gibberella zeae with their EC50s falling between 4.0 and 5.5 μg/mL, while YZK-C22 showed an EC50 of 6.4 μg/mL. In addition, 5ah exhibited promising in vivo activity against Erysiphe graminis and Puccinia sorghi Schw. with 100% efficacy at 10 μg/mL and 90% efficacy at 2 μg/mL against P. sorghi Schw. Compound 5ai showed good PK inhibitory activity with an IC50 of 38.8 μmol/L, and it was well docked into the active site of the target enzyme PK, which was slightly more active than YZK-C22 with its IC50 of 42.4 μmol/L. Our studies discovered that isothiazole-purines were PK-based fungicidal leads deserving of further study.

Natural products are one of the resources for discovering novel fungicidal leads. As a natural fungicide, osthole was used as a coumarin-based lead compound for the development of novel fungicides. Here, a series of 3,4-dichloroisothiazole-containing 7-hydroxycoumarins were rationally designed, synthesized, and characterized by introducing a bioactive substructure, 3,4-dichloroisothiazole, into the coumarin skeleton. In vitro bioassay indicated that compound 7g displayed good activity against Rhizoctonia solani, Physalospora piricola, Sclerotinia sclerotiorum, and Botrytis cinerea. Its median effective concentration (EC50) value against each of these fungi fell between 0.88 and 2.50 μg/mL, which was much lower than that of osthole against the corresponding pathogen (between 7.38 and 74.59 μg/mL). In vivo screening validated that 7k exhibited 100%, 60%, and 20% efficacy against R. solani Kuhn at 200, 100, and 50 μg/mL, respectively. RNA sequence analysis implied that growth inhibition of R. solani by 7k might result from potential disruptions of fungal membrane formation and intracellular metabolism. Furthermore, a field experiment with cucumber plants indicated that 7b showed 62.73% and 74.03% efficacy against Pseudoperonospora cubensis (Berk. & Curt.) Rostov. at rates of 12.5 g a.i./ha and 25 g a.i./ha, respectively, which showed no significant difference between 7b and osthole at 30 g a.i./ha. Our studies suggested that 7b, 7g, and 7k might be used as fungicidal leads for further optimization.

In recent years, the avian influenza has brought not only serious economic loss to the poultry industry in China but also a serious threat to human health because of the avian influenza virus (AIV) gene recombination and reassortment. Until now, traditional RT-PCR, fluorescence RT-PCR and virus isolation identification have been developed and utilized to detect AIV, but these methods require high-level instruments and experimental conditions, not suitable for the rapid detection in field and farms. In order to develop a rapid, sensitive and practical method to detect and identify AIV subtypes, 4 specific primers to the conserved region of AIV M gene were designed and a loop-mediated isothermal amplification (RT-LAMP) method was established. Using this method, the M gene of H1–H16 subtypes of AIV were amplified in 30 min with a water bath and all 16 H subtypes of AIV were able to be visually identified in presence of fluorescein, without cross reaction with other susceptible avian viruses. In addition, the detection limit of the common H1, H5, H7, and H9 AIV subtypes with the RT-LAMP method was 0.1 PFU (plaque-forming unit), which was 10 times more sensitive than that using the routine RT-PCR. Further comparative tests found that the positivity rate of RT-LAMP on detecting clinical samples was 4.18% (14/335) comparing with 3.58% (12/335) from real-time RT-PCR. All these results suggested that the RT-LAMP method can specifically detect and identify AIV with high sensitivity and can be considered as a fast, convenient and practical method for the clinic test and epidemiological investigation of AIV.

The melanocortin-3 receptor (MC3R) is known to be involved in regulation of energy homeostasis, regulating feed efficiency and nutrient partitioning in mammals. Its physiological roles in non-mammalian vertebrates, especially economically important aquaculture species, are not well understood. Channel catfish (Ictalurus punctatus) is the main freshwater aquaculture species in North America. In this study, we characterized the channel catfish MC3R. The mc3r of channel catfish encoded a putative protein (ipMC3R) of 367 amino acids. We transfected HEK293T cells with ipMC3R plasmid for functional studies. Five agonists, including adrenocorticotropin, α-melanocyte stimulating hormone (α-MSH), β-MSH, [Nle4, D-Phe7]-α-MSH, and D-Trp8-γ-MSH, were used in the pharmacological studies. Our results showed that ipMC3R bound β-MSH with higher affinity and D-Trp8-γ-MSH with lower affinity compared with human MC3R. All agonists could stimulate ipMC3R and increase intracellular cAMP production with sub-nanomolar potencies. The extracellular signal-regulated kinases 1 and 2 (ERK1/2) activation could also be triggered by ipMC3R. The ipMC3R exhibited constitutive activities in both cAMP and ERK1/2 pathways, and Agouti-related protein served as an inverse agonist at ipMC3R, potently inhibiting the high basal cAMP level. Moreover, we showed that melanocortin receptor accessory protein 2 (MRAP2) preferentially modulated ipMC3R in cAMP production rather than ERK1/2 activation. Our study will assist further investigation of the physiological roles of the ipMC3R, especially in energy homeostasis, in channel catfish.

Increasing atmospheric carbon dioxide (ab. CO₂) and fertilizer‐nitrogen (ab. N) applications may have marked direct effects on the plant growth of agricultural crops, and in turn affect the higher trophic level of insect herbivores. In this study, the effects of elevated CO₂ (i.e., 650 µl/L vs. ambient 400 µl/L) and fertilizer‐N (0, 50, 100, 200 kg/ha) on the population abundances and the inter‐specific competition among three co‐occurring species of wheat aphids, Sitobion avenae, Rhopalosiphum padi and Schizaphis graminum, were studied. The grain weight per ear and the 1,000‐grain weight were generally increased when grown under elevated CO₂ and showed a significant effect at the 100 kg/ha (grain weight per ear) and 0, 50 and 100 kg/ha (1,000‐grain weight) N. These two yield indexes increased with increasing fertilizer‐N levels within reasonable limits and reached a maximum at 100 kg/ha. Elevated CO₂ combined with fertilizer‐N levels formed complex indirect effects on the three wheat aphids through the wheat crops they fed on. Elevated CO₂ significantly decreased the niche overlap index (ab. NOI) between S. avenae and R. padi under 0 and 100 kg/ha and that between R. padi and S. graminum under 0 kg/ha, while significantly increased the three NOIs under 50 kg/ha and that between R. padi and S. graminum under 100 and 200 kg/ha. S. avenae and R. padi had the larger population and stronger competition in low‐N condition (0 and 50 kg/ha), which was harmful to wheat yield and quality when combined with its own poor nutrition. Overall, the 100 kg/ha N level was the best option based on the aphid population, competition and wheat yields. Therefore, the balanced relationship formed among fertilizers, plants and insects under 100 kg/ha N was vital for the interactive ecosystem.

Plants adjust amplitude and duration of immune responses via different strategies to maintain growth, development, and resistance to pathogens. Pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI) play vital roles. Pattern recognition receptors, comprising a large number of receptor-like protein kinases and receptor-like proteins, recognize related ligands and trigger immunity. PTI is the first layer of the innate immune system, and it recognizes PAMPs at the plasma membrane to prevent infection. However, pathogens exploit effector proteins to bypass or directly inhibit the PTI immune pathway. Consistently, plants have evolved intracellular nucleotide-binding domain and leucine-rich repeat-containing proteins to detect pathogenic effectors and trigger a hypersensitive response to activate ETI. PTI and ETI work together to protect plants from infection by viruses and other pathogens. Diverse receptors and the corresponding ligands, especially several pairs of well-studied receptors and ligands in PTI immunity, are reviewed to illustrate the dynamic process of PTI response here. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Twenty-one novel 3,4-dichloroisothiazolocoumarin-containing strobilurins were rationally designed and synthesized. Preliminary bioassay showed that compounds 7c, 7h and 7l exhibited over 80% inhibitory rate against Sclerotinia sclerotiorum at 50 μg/mL, 7c exhibited good activity against S. sclerotiorum with median effective concentration (EC50) of 4.08 μg/mL, while the positive control coumoxystrobin showed EC50 of 1.00 μg/mL. In addition, 7d showed better fungicidal activity with a lower EC50 value of 7.65 μg/mL against Botrytis cinerea than that of positive control trifloxystrobin with its EC50 value of 21.96 μg/mL. This study indicated that 3,4-dichloroisothiazolocoumarin-containing strobilurin was a promising fungicide lead deserved for further study.

Firstly, the basic conditions of the fungicide were introduced, its action principle was analyzed, the use method of the fungicide in the military warehouse was put forward, and the factors influencing the application effect were discussed.

This study investigated the effects of spontaneous and Lactobacillus plantarum (LP) fermentation on structure glutinous proso millet starch (GPM) and physical properties. In order to explore starch changes by fermentation for quality regulation of fermented food glutinous proso millet and improve properties of modified GPM starch products. The molecular weight, amylopectin chain length distribution, morphological properties, short-range ordered structure, and crystal structure of GPM starch were analyzed. Results showed that compared with the non-fermented GPM starch, amylose content of spontaneous and LP-fermented GPM starch increased then decreased, with the latter decreasing faster. There were no changes in the Type A structures of spontaneous fermented, LP-fermented, and non-fermented GPM starch. After 24 h of LP fermentation, higher water swelling power, and lower GPM starch solubility were obtained compared to non-fermentation and spontaneous fermentation. The decrease in pasting properties and thermal properties while increasing GPM starch properties was shown after fermentation compared to non-fermentation experiments. Besides, LP fermentation showed faster changes of these properties than spontaneous fermentation. In conclusion, our findings showed that fermentation could improve GPM starch properties, benefiting fermented GPM products and as a modification method for GPM starch. Especially, LP fermentation was more effective than spontaneous fermentation.

MicroRNAs (miR) are single-stranded RNA of 21-23 nucleotides in length that repress mRNA translation and induces mRNA degradation. miR acts as an endogenous factor of gene expression and plays a crucial part in cancer biology such as cell development, proliferation, differentiation, and apoptosis. Numerous research has indicated that dysregulation of miR associates with colorectal carcinogenesis. In this review article, we firstly introduce the background of miR and colorectal cancer, and the mechanisms of miR in colorectal cancer, such as the proliferation, apoptosis, and progression. Then, we summarize the theranostic value of miR in colorectal cancer. Eventually, we discuss the potential directions and perspectives of miR. This article serves as a guide for further studies and implicate miR as a potent theranostic target for colorectal cancer.

ObjectivesDespite preclinical studies involving miRNA therapeutics conducted in osteoarthritis (OA) over the years, none of these miRNAs have yet translated to clinical applications, owing largely to the lack of efficient intra-articular (IA) delivery systems. Here, we investigated therapeutic efficacy of the chondrocyte-specific aptamer-decorated PEGylated polyamidoamine nanoparticles (NPs)-based miRNAs delivery for OA.MethodsThe role of miR-141/200c cluster during skeletal and OA development was examined by miR-141/200cflox/floxmice and Col2a1-CreERT2; miR-141/200cflox/floxmice. Histological analysis was performed in mouse joints and human cartilage specimens. Chondrocyte-specific aptamer-decorated NPs was designed, and its penetration, stability and safety were evaluated. OA progression was assessed by micro-CT analysis, X-ray and Osteoarthritis Research Society International scores after destabilising the medial meniscus surgery with miR-141/200c manipulation by NPs IA injection. Mass spectrometry analysis, molecular docking and molecular dynamics simulations were performed to investigate the interaction between aptamer and receptor.ResultsIncreased retention of NPs inside joint space is observed. The NPs are freely and deeply penetrant to mice and human cartilage, and unexpectedly persist in chondrocytes for at least 5 weeks. OA chondrocytes microenviroment improves endo/lysosomal escape of microRNAs (miRNAs). Therapeutically, IA injection of miR-141/200c inhibitors provides strong chondroprotection, whereas ectopic expression of miR-141/200c exacerbates OA. Mechanistically, miR-141/200c promotes OA by targeting SIRT1, which acetylates histone in the promoters of interleukin 6 (IL-6), thereby activating IL-6/STAT3 pathway.ConclusionsOur findings indicate that this nanocarrier can optimise the transport kinetics of miR-141/200c into chondrocytes, fostering miRNA-specific disease-modifying OA drugs development.

G protein-coupled receptors (GPCRs) play critical roles in transmitting a variety of extracellular signals into the cells and regulate diverse physiological functions. Naturally occurring mutations that result in dysfunctions of GPCRs have been known as the causes of numerous diseases. Significant progresses have been made in elucidating the pathophysiology of diseases caused by mutations. The multiple intracellular signaling pathways, such as G protein-dependent and β-arrestin-dependent signaling, in conjunction with recent advances on biased agonism, have broadened the view on the molecular mechanism of disease pathogenesis. This review aims to briefly discuss biased agonism of GPCRs (biased ligands and biased receptors), summarize the naturally occurring GPCR mutations that cause biased signaling, and propose the potential pathophysiological relevance of biased mutant GPCRs associated with various endocrine diseases.

Naphthenic acids (NAs) are cycloalkyl linear carboxylic acids primarily originating from oil sand tailings water (OSTW), and the exposure to NAs affects the photosynthesis of marine microalgaes. However, the transcriptional response upon the stress of NAs remains unclear. Herein, we investigate the transcriptome alterations of marine Phaeodactylum tricornutum induced by NAs using RNA-sequencing. In total 70 differentially expressed genes (DEGs) are enriched in seven biological process pathways, and 48 DEGs are enriched in four molecular functions. Differentially expressed genes of P. tricornutum are primarily concentrated in three photosynthesis-related pathways including the porphyrin and chlorophyll metabolism pathway, the photosynthesis pathway, and the carbon fixation pathway in photosynthetic organisms. Five DEGs such as hemE, chlM, TPI/GapC3, FbaC5, and CHLH are validated using quantitative real-time PCR assay. NAs at low concentrations affect the material energy conversion efficiency of P. tricornutum, such as chlorophyll synthesis, electron transfer efficiency, and carbon assimilation.

Four new triterpene glucosides (1–4) were isolated from the 90% ethanol extract of Salacia cochinchinensis, together with five known compounds (5–9). The structures of the new compounds were elucidated by comprehensive spectroscopic analysis including HRESIMS, IR, 1 D and 2 D NMR analysis. All isolates were assayed for their α-glucosidase inhibitory activity. Compound 9 showed remarkable α-glucosidase inhibitory activity with an IC50 value of 0.31 μM, and the triterpene glycosides (1–5) exhibited moderate α-glucosidase inhibitory activity.

Plant vacuolar H+-transporting inorganic pyrophosphatase (V-PPase; EC 3.6.1.1) is a crucial enzyme that exists on the tonoplast to maintain pH homeostasis across the vacuolar membrane. This enzyme generates proton gradient between cytosol and vacuolar lumen by hydrolysis of a metabolic byproduct, pyrophosphate (PP i ). The regulation of V-PPase at protein level has drawn attentions of many workers for decades, but its mechanism is still unclear. In this work, we show that AVP1, the V-PPase from Arabidopsis thaliana, is a target protein for regulatory 14-3-3 proteins at the vacuolar membrane, and all twelve 14-3-3 isoforms were analyzed for their association with AVP1. In the presence of 14-3-3ν, -µ, -ο, and -ι, both enzymatic activities and its associated proton pumping of AVP1 were increased. Among these 14-3-3 proteins, 14-3-3 µ shows the highest stimulation on coupling efficiency. Furthermore, 14-3-3ν, -µ, -ο, and -ι exerted protection of AVP1 against the inhibition of suicidal substrate PP i at high concentration. Moreover, the thermal profile revealed the presence of 14-3-3ο improves the structural stability of AVP1 against high temperature deterioration. Additionally, the 14-3-3 proteins mitigate the inhibition of Na+ to AVP1. Besides, the binding sites/motifs of AVP1 were identified for each 14-3-3 protein. Taken together, a working model was proposed to elucidate the association of 14-3-3 proteins with AVP1 for stimulation of its enzymatic activity.

Mesenchymal stem cells (MSCs) may be easily isolated from the bone marrow, and possess multi-lineage differentiation potential and various therapeutic applications. The differentiation of MSCs into osteoblasts is a complex process that is regulated by multiple internal and external factors. In the present study, the differentiation of MSCs isolated from rabbit bone marrow into osteoblasts using different osteoblast inductive media in the presence of dexamethasone, bone morphogenetic protein 2 (BMP-2), 1,25-dihydroxyvitamin D3, transforming growth factor β (TGFβ), platelet lysate and cyclooxygenase 2 (COX2), respectively. Alkaline phosphatase (ALP) activity, mineralization, collagen type (Ct) I and osteocalcin activities, and the mRNA and protein expression levels of vascular endothelial growth factor (VEGF), BMP-2 and Ct II were measured during the differentiation process in MSCs treated with different inducers. Rabbit MSCs were successfully isolated and were observed to be predominantly circular in shape after culture for 24 h. Following subculture for 5 days, the cells demonstrated a spindle shape. ALP, Ct I and osteocalcin activities were higher in cells cultured in dexamethasone, BMP-2 and TGFβ compared with the activities in control cells. Following differentiation, the dexamethasone, BMP-2 and TGFβ groups demonstrated significantly enhanced mineralization of MSCs detected by Alizarin Red S staining. The mRNA and protein expression levels of VEGF, BMP-2 and Ct II were significantly increased in the same groups compared with the levels in the control group. In conclusion, rabbit MSCs were successfully isolated from bone marrow and differentiated into osteoblasts indicated by raised ALP, Ct I and osteocalcin activities, mineralization and expression of osteogenesis-inducing genes and proteins. The present study revealed that dexamethasone, BMP-2 and TGFβ have a positive effect on cell differentiation.

The global prevalence of obesity highlights the importance of understanding on regulation of energy homeostasis. The central melanocortin system is an important intersection connecting the neural pathways controlling satiety and energy expenditure to regulate energy homeostasis by sensing and integrating the signals of external stimuli. In this system, neural melanocortin receptors (MCRs), melanocortin-3 and -4 receptors (MC3R and MC4R), play crucial roles in the regulation of energy homeostasis. Recently, multiple intracellular signaling pathways and biased signaling at neural MCRs have been discovered, providing new insights into neural MCR signaling. This review attempts to summarize biased signaling including biased receptor mutants (both naturally occurring and lab-generated) and biased ligands at neural MCRs, and to provide a better understanding of obesity pathogenesis and new therapeutic implications for obesity treatment.

Glioma growth and progression are characterized by abundant development of blood vessels that are highly aberrant and poorly functional, with detrimental consequences for drug delivery efficacy. The mechanisms driving this vessel dysmorphia during tumor progression are poorly understood. Using longitudinal intravital imaging in a mouse glioma model, we identify that dynamic sprouting and functional morphogenesis of a highly branched vessel network characterize the initial tumor growth, dramatically changing to vessel expansion, leakage, and loss of branching complexity in the later stages. This vascular phenotype transition was accompanied by recruitment of predominantly pro-inflammatory M1-like macrophages in the early stages, followed by in situ repolarization to M2-like macrophages, which produced VEGF-A and relocate to perivascular areas. A similar enrichment and perivascular accumulation of M2 versus M1 macrophages correlated with vessel dilation and malignancy in human glioma samples of different WHO malignancy grade. Targeting macrophages using anti-CSF1 treatment restored normal blood vessel patterning and function. Combination treatment with chemotherapy showed survival benefit, suggesting that targeting macrophages as the key driver of blood vessel dysmorphia in glioma progression presents opportunities to improve efficacy of chemotherapeutic agents. We propose that vessel dysfunction is not simply a general feature of tumor vessel formation, but rather an emergent property resulting from a dynamic and functional reorganization of the tumor stroma and its angiogenic influences. ispartof: EMBO Molecular Medicine vol:9 issue:12 pages:1629-1645 ispartof: location:England status: published

Background The current published prognosis models for brain metastases (BMs) from cancer have not addressed the issue of either newly diagnosed non–small-cell lung cancer (NSCLC) with BMs or the lung cancer genotype. We sought to build an adjusted prognosis analysis (APA) model, a new prognosis model specifically for NSCLC patients with BMs at the initial diagnosis using adjusted prognosis analysis (APA). Patients and Methods The model was derived using data from 1158 consecutive patients, with 837 in the derivation cohort and 321 in the validation cohort. The patients had initially received a diagnosis of BMs from NSCLC at Sun Yat-Sen University Cancer Center from 1994 to 2015. The prognostic factors analyzed included patient characteristics, disease characteristics, and treatments. The APA model was built according to the numerical score derived from the hazard ratio of each independent prognostic variable. The predictive accuracy of the APA model was determined using a concordance index and was compared with current prognosis models. The results were validated using bootstrap resampling and a validation cohort. Results We established 2 prognostic models (APA 1 and 2) for the whole group of patients and for those with known epidermal growth factor receptor (EGFR) genotype, respectively. Six factors were independently associated with survival time: Karnofsky performance status, age, smoking history (replaced by EGFR mutation in APA 2), local treatment of intracranial metastases, EGFR-tyrosine kinase inhibitor treatment, and chemotherapy. Patients in the derivation cohort were stratified into low- (score, 0-2), moderate- (score, 3-5), and high-risk (score 6-7) groups according to the median survival time (16.6, 10.3, and 5.2 months, respectively; P Conclusion Compared with recursive partition analysis and graded prognostic assessment, APA seems to be more suitable for initially diagnosed NSCLC with BMs.

Background Glutathione S-transferase omega 1 (GSTO1), as a member of the glutathione S-transferase (GST) family genes, has been discovered to be up-regulated in several cancer cell lines which exhibited strong aggressiveness. However, the function of GSTO1 on cutaneous malignant melanoma (CMM) has not been illuminated. Methods Outcome of expression level and prognosis of GSTO1 were obtained from Oncomine and TCGA database. The specific effects of GSTO1 on the characteristics and regulatory mechanism of CMM cells were demonstrated by cell counting kit-8, colony formation, flow cytometry, and transwell assays in vitro. Western blot was employed to analyze the expression of proliferating cell nuclear antigen (PCNA), p53 and epithelial-to-mesenchymal (EMT) related proteins. Results We observed that GSTO1 was up-regulated in CMM samples when compared with the corresponding controls. Moreover, patients in CMM with high expression of GSTO1 were more likely to have a poor prognosis. Through in vitro experiments, silenced GSTO1 resulted in inhibition of CMM cells growth and aggressiveness, increased cell apoptosis, and blocked cell cycle. Finally, the expression of PCNA, p53 and EMT-related proteins were changed due to reduction of GSTO1. Conclusions To sum up, our outcomes exhibited that weakening GSTO1 reduced the proliferation and mobility of CMM cells, increased the apoptosis ability of CMM cells, and arrested cell cycle at G1 phase, which can be achieved by affecting the expression of PCNA, p53 and the EMT process. This discovery provided a new perspective for elucidating the mechanism of CMM, and offered theoretical support for searching clinical therapeutic targets in the future.

Mesenchymal stem cells (MSC) have the potential of multi-directional differentiation, have stable genetic information, can self-replicate and renew, and are commonly used as seed cells in the field of regeneration1. Adipose mesenchymal stem cells, bone marrow mesenchymal stem cells and umbilical cord mesenchymal stem cells are the most commonly used mesenchymal stem cells. Compared with other mesenchymal stem cells, adipose mesenchymal stem cells have a wide range of sources, relatively simple acquisition methods and less trauma2. Therefore, adipose mesenchymal stem cells have gradually become a new favorite in stem cell research. However, due to the harsh storage conditions and inconvenient transportation of living cells, the survival rate of direct transplantation into living organisms is low, which makes it difficult for clinical application. Exosome is a kind of extracellular secretory vesicle, which exists in almost all body fluids and can be extracted from the supernatant of most cultured cells in vitro3. It has the characteristics of secretory cells, simple storage, convenient transportation and low immunogenicity, providing a new direction for the clinical application of adipose mesenchymal stem cells. Therefore, this paper reviews the exosomes derived from adipose mesenchymal stem cells.Read Complete Article at ijSciences: V82019021984 AND DOI: http://dx.doi.org/10.18483/ijSci.1984

The system of hepatocyte growth factor (HGF) and its receptor c‐Met plays a critical role in tumor invasive growth and metastasis. The mortality rate of colorectal cancer (CRC), one of the most commonly diagnosed malignancies, is increased by it gradual development into metastasis, most frequently in the liver. Overexpression of c‐Met, the protein tyrosine kinase receptor for the HCF/scatter factor, has been implicated in the progression and metastasis of human colorectal carcinoma. In this study, we aimed to investigate the role of c‐Met in CRC liver metastasis and illustrate the clinical impact of regulating HGF/c‐Met signaling in patients with CRC liver metastasis. We found that (I) higher levels of c‐Met expression (mRNA and Protein) in CRC liver metastasis than primary CRC by assessing the patient tissue samples; (II) a positive correlation of c‐Met expression with tumor stages of CRC liver metastasis, as well as c‐Met expression in CRC, live metastasis concurred with regional lymph node metastasis; (III) the clinical impact of downregulation of HGF/c‐Met signaling on the reduction of proliferation and invasion in CRC liver metastasis. Therefore, we demonstrate that the regulation of HGF/c‐Met pathways may be a promising strategy in the treatment of patients with CRC liver metastasis.

The authors describe the preparation of gold-platinum nanoflower (AuPt NFs) and show that they can be simultaneously used as a label and as an enzyme mimic in lateral flow immunoassays (LFIs). The AuPt NFs were prepared by growing Pt nanowires on the surface of gold nanoparticle. The assay involves the capture of target proteins (here: rabbit IgG as a model analyte) by the immobilized capture antibody, and by using AuPt NF-labeled secondary antibody. The AuPt NFs are thus captured by the test zone and produce a characteristic black band for visual detection of the antigen (IgG). The coloration of the test line can be further enhanced by addition of the chromogenic substrate 3-amino-9-ethyl-carbazole which is catalytically oxidized by the captured Pt nanowires on the AuPt NF and produce a red coloration. Quantitative results were obtained by reading the test line intensities with a portable strip reader. The LFI has a 5 pg mL-1 detection limit for IgG under optimized experimental conditions. This is 100 times lower than that of the conventional AuNP-based LFI. Conceivably, this assay has a wide scope in that it may be applied to numerous other targets for which appropriate antibodies are available. Graphical abstract Gold-platinum nanoflowers are used as a label and as an enzyme mimic in a highly sensitive lateral flow immunoassay for IgG. The detection limit of gold-platinum nanoflower-based lateral flow assay is 100 times lower than that of the conventional gold nanopaticle-based lateral flow assay.

Metastasis is the most common cause of death in ovarian cancer patients but remains largely untreated. Epithelial‑mesenchymal transition (EMT) is critical for the conversion of early‑stage ovarian tumors into metastatic malignancies. Thus, investigating the signaling pathways promoting EMT may identify potential targets for the treatment of metastatic ovarian cancer. Lysine demethylase 2A (KDM2A), also known as FBXL11 and JHDM1A, is a histone H3 lysine 36 (H3K36) demethylase that regulates EMT and the metastasis of ovarian cancer. However, the function and underlying mechanisms of EMT suppression in ovarian cancer have not been thoroughly elucidated to date. In the present study, we used Gene Expression Omnibus (GEO) databases to determine that KDM2A is significantly upregulated in human ovarian cancers. KDM2A expression was assessed by immunohistochemistry of epithelial ovarian cancer (EOC) borderline ovarian tumors and normal ovary tissues. Seven fresh EOC tissues and 3 fresh normal ovary tissues were collected for western blot analysis. Kaplan‑Meier survival curves were constructed to identify genes related to EOC prognosis from the TCGA data portal. Stable KDM2A‑knockdown cell lines were established to study the biological functions and underlying mechanisms of KDM2A in EMT in vitro. GEO database analysis revealed that KDM2A was highly upregulated in EOC tissues; this analysis was accompanied by immunochemistry and western blot analysis using samples of human tissues. High expression of KDM2A was associated with poor survival in EOC patients. KDM2A knockdown promoted apoptosis and suppressed the proliferation, migration and invasion of tumor cells in vitro. EMT and the PI3K/AKT/mTOR signaling pathway were suppressed in KDM2A‑silenced cells. Inactivation of the PI3K/AKT/mTOR signaling pathway in A2780 cells induced EMT inhibition. Our data revealed that KDM2A functions as a tumor oncogene, and the downregulation of KDM2A expression regulates EMT and EOC progression, providing a valuable prognostic marker and potential target for the treatment of EOC patients.

Stone cells negatively affect fruit quality because of their firm and lignified cell walls, so are targets for reduction in pear breeding programmes. However, there is only limited knowledge of the molecular mechanisms underlying the formation of stone cells. Here, we show that PbrMYB169, an R2R3 MYB transcription factor, of Pyrus bretschneideri positively regulates lignification of stone cells in pear fruit. PbrMYB169 was shown to be co-expressed with lignin biosynthesis genes during pear fruit development, and this co-expression pattern was coincident with stone cell formation in the fruit of Pyrus bretschneideri ‘Dangshansuli’. The PbrMYB169 expression level was also positively correlated with stone cell content in 36 pear cultivars tested. PbrMYB169 protein significantly activated the promoter of lignin genes C3H1, CCR1, CCOMT2, CAD, 4CL1, 4CL2, HCT2, and LAC18 via binding to AC elements [ACC(T/A)ACC] in these promoters. Furthermore, overexpression of PbrMYB169 in transgenic Arabidopsis plants enhanced the expression of lignin genes, and increased lignin deposition and cell wall thickness of vessel elements, but did not change the ratio of syringyl and guaiacyl lignin monomers. In conclusion, PbrMYB169 appears to be a transcriptional activator of lignin biosynthesis and regulates secondary wall formation in fruit stone cells. This study advances the understanding of the regulation of lignin biosynthesis and provides valuable molecular genetic information for reducing stone cell content in pear fruit.

Provenance selection plays a major role for the afforestation success, resilience and persistence of forests into the future. Understanding the germination response to water potential and temperature among provenances is crucial for forest restoration initiatives. However, afforestation programs often neglect the seed germination traits from a selected provenance, threatening natural or artificial regeneration in future. In this study, we assessed the impact of water potential (0 and -0.2 MPa) and temperature (20, 25, and 30°C) on germination of Pinus yunnanensis seeds from 15 provenances. We found a clear effect of seed provenance on the germination percentage and mean germination time. In general, the lower water potential (-0.2 MPa) led to a reduction of germination percentage for most provenances, especially at 30°C; and the higher temperature (30°C) and water potential (0 MPa) resulted in low mean germination time. Germination percentage and mean germination time responses to water potential and temperature varied among provenances. We observed that germination percentage was significantly positively correlated with annual mean temperature, mean temperature of driest quarter and seed mass associated with the provenance, but significantly negatively correlated with annual mean precipitation, and mean precipitation of driest quarter and wettest quarter. Significant relationships between mean germination time and those variables were contrary to germination percentage. However, the significance of these relationships varied in all the treatments. Therefore, we conclude that germination of P. yunnanensis seeds from different provenances adapt diversely to abiotic stresses; this is likely to be important for provenance selection in future reforestation. Our results highlight that future re-forestation efforts need to critically evaluate larger numbers of provenances to understand thresholds limiting performance and establishment success.

Ethnopharmacological relevance Atractylodis Rhizoma (AR), mainly includes Atractylodes lancea (Thunb.) DC. (A. lancea) and Atractylodes chinensis (DC.) Koidz. (A. chinensis) is widely used in East Asia as a diuretic and stomachic drug, for the treatment of rheumatic diseases, digestive disorders, night blindness, and influenza as it contains a variety of sesquiterpenoids and other components of medicinal importance. Aim of the review A systematic summary on the botany, traditional uses, phytochemistry, pharmacology, toxicology, and quality control of AR was presented to explore the future therapeutic potential and scientific potential of this plant. Materials and methods A review of the literature was performed by consulting scientific databases including Google Scholar, Web of Science, Baidu Scholar, Springer, PubMed, ScienceDirect, CNKI, etc. Plant taxonomy was confirmed to the database “The Plant List”. Results Over 200 chemical compounds have been isolated from AR, notably sesquiterpenoids and alkynes. Various pharmacological activities have been demonstrated, especially improving gastrointestinal function and thus allowed to assert most of the traditional uses of AR. Conclusions The researches on AR are extensive, but gaps still remain. The molecular mechanism, structure-activity relationship, potential synergistic and antagonistic effects of these components need to be further elucidated. It is suggested that further studies should be carried out in the aspects of comprehensive evaluation of the quality of medicinal materials, understanding of the “effective forms” and “additive effects” of the pharmacodynamic substances based on the same pharmacophore of TCM, and its long-term toxicity in vivo and clinical efficacy., Graphical abstract Image 1

Objective Disordered programmed cell death may play a role in the development of superficial venous incompetence. We have determined the number of cells undergoing apoptosis and the alterations in the apoptotic level in the wall of different segments of the great saphenous varicose vein. Methods Twenty-one varicose great saphenous veins (VGSVs) (varicose group) and 12 normal great saphenous veins (GSVs) (control group) were collected, and the apoptosis level in the upper, middle, and lower segments were immunohistochemically stained with antibodies (anti-Bax and anti-Bcl-xl). Apoptosis was evaluated by the TUNEL assay and immunofluorescence staining. The morphology of apoptotic cells was observed with an electron microscope. Results Quantitative analysis showed that the apoptotic ratios in venous walls (intima and media) of the varicose group were significantly lower than the corresponding regions in the control group (all P Conclusion VGSV walls were found to have a significant decrease in apoptotic rate compared with that of GSVs. The rate of apoptosis in the intima and media within the upper segment was increased more than the middle and lower segments in the GSVs. Our findings confirm that programmed cell death is down-regulated in primary varicose veins.

// Li-Kun Hou 1, 2, 3, * , Yue Yu 1, 2, 3, * , Ye-Gong Xie 1, 2, 3 , Jie Wang 1, 2, 3 , Jie-Fei Mao 1, 2, 3 , Bin Zhang 1, 2, 3 , Xin Wang 1, 2, 3 , Xu-Chen Cao 1, 2, 3 1 The First Department of Breast Cancer, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin 300060, China 2 Key Laboratory of Cancer Prevention and Therapy, Tianjin 300060, China 3 Key Laboratory of Breast Cancer Prevention and Therapy, Tianjin Medical University, Ministry of Education, Tianjin 300060, China * These authors contributed equally to this work Correspondence to: Xu-Chen Cao, e-mail: caoxuchen@tmu.edu.cn Keywords: miR-340, ZEB1, TGF-β, EMT, breast cancer Received: October 27, 2015 Accepted: March 06, 2016 Published: March 27, 2016 ABSTRACT MicroRNAs act as key regulators in carcinogenesis and progression in various cancers. In present study, we explored the role of miR-340 in the breast cancer progression. Our results showed that overexpression of miR-340 inhibits breast cancer cell proliferation and invasion, whereas depletion of miR-340 promotes breast cancer progression. Molecularly, ZEB1 was identified as a target gene of miR-340 and miR-340 suppressed the expression of ZEB1 by directly binding to the 3′-UTR of ZEB1. Furthermore, ZEB1 transcriptionally suppresses miR-340 expression. The negative feedback loop regulated TGF-β-mediated breast cancer progression. In conclusion, our data suggested that miR-340 acted as a tumor suppressor in breast cancer progression.

Serum deprivation response (SDPR), a key substrate for protein kinase C, play a critical role in inducing membrane curvature and participate in the formation of caveolae. However, the function of SDPR in cancer development and progression is still not clear. Here, we found that SDPR is downregulated in human breast cancer. Overexpression of SDPR suppresses cell proliferation and invasion in MDA‐MB‐231 cells, while depletion of SDPR promotes cell proliferation and invasion in MCF10A cells. Subsequently, SDPR depletion induces epithelial–mesenchymal transition (EMT)‐like phenotype. Finally, knockdown of SDPR activates transforming growth factor‐β (TGF‐β) signaling by upregulation of TGF‐β1 expression. In conclusion, our results showed that SDPR inhibits breast cancer progression by blocking TGF‐β signaling. Serum deprivation response suppresses cell proliferation and invasion in breast cancer cells. SDPR depletion induces epithelial–mesenchymal transition by activation of TGF‐β signaling.

Objective: Green Paulownia (hybridization of Paulownia elongata × Paulownia fotunei and tropical Paulownia spp.) is new hybrid claimed as one of the fast-growing woody plants with the high potential as a fiber material or lignocellulosic material. The material for this study originates from the area of Nanning in China. Methodology: Cell morphology and anatomical appearances were observed and evaluated under the image analysis system (Leica DMLS). Physical and mechanical properties were evaluated based on the American Society for Testing and Materials (ASTM) standards. Results: From the results, average value of the mean fiber length was 0.905 mm, mean fiber length 34.59 μm, lumen thickness 26.80 μm and cell wall thickness 3.89 μm. Fiber dimensions of green Paulownia are in the normal range for hardwoods. The physical and mechanical properties of 3 years old green Paulownia have similar properties than those 7-11 years old Paulownia published in China. Conclusion: The 3 years old green Paulownia timbers can be used as materials for furniture.

Recent studies indicated that bisphenol A (BPA) can disrupt spermatogenesis and then cause male infertility. The present study revealed that BPA greater than 10-6 M inhibited the proliferation of Leydig TM3 cells via a concentration dependent manner. The proteomic study revealed that 50 proteins were modulated in TM3 cells following exposure to BPA, which was relevant to structure, motility, cell metabolism, protein and nucleotide processing, and cell proliferation. Furthermore, BPA increased the in vitro migration and invasion of Leydig TM3 cells, which might be due to the BPA's modulation of proteins related to cell structure and motility such as actin and heat shock protein (HSP). Silencing of galectin-1, which was up regulated by BPA, significantly abolished the BPA-induced migration of TM3 cells. BPA treatment obviously increased the phosphorylation of ERK1/2 and Akt, while only PD98509 (ERK1/2 inhibitor) significantly attenuated BPA induced up regulation of galectin-1. Furthermore, PD98509 also reversed BPA induced migration of TM3 cells. Our study demonstrated that xenoestrogen BPA at micromolar or greater concentrations can modulate protein profiles, inhibit cell proliferation, and promote the in vitro migration and invasion of Leydig TM3 cells. It provided new insight into the mechanisms responsible for BPA induced male infertility.

Melanocortin-4 receptor (MC4R) plays critical roles in the regulation of various physiological processes, such as energy homeostasis, reproduction and sexual function, cardiovascular function, and other functions in mammals. Although the functions of the MC4R in fish have not been extensively studied, the importance of MC4R in regulation of piscine energy expenditure and sexual functions is emerging. Swamp eel (Monopterus albus) is an economically and evolutionarily important fish widely distributed in tropics and subtropics. We cloned swamp eel mc4r (mamc4r), consisting of a 981 bp open reading frame encoding a protein of 326 amino acids. The sequence of maMC4R was homologous to those of several teleost MC4Rs. Phylogenetic and chromosomal synteny analyses showed that maMC4R was closely related to piscine MC4Rs. qRT-PCR revealed that mc4r transcripts were highly expressed in brain and gonads of swamp eel. The maMC4R was further demonstrated to be a functional receptor by pharmacological studies. Four agonists, α-melanocyte stimulating hormone (α-MSH), β-MSH, [Nle4, D-Phe7]-α-MSH (NDP-MSH), and adrenocorticotropin, could bind to maMC4R and induce intracellular cAMP production dose-dependently. Small molecule agonist THIQ allosterically bound to maMC4R and exerted its effect. Similar to other fish MC4Rs, maMC4R also exhibited significantly increased basal activity compared with that of human MC4R. The high basal activity of maMC4R could be decreased by inverse agonist ML00253764, suggesting that maMC4R was indeed constitutively active. The availability of maMC4R and its pharmacological characteristics will facilitate the investigation of its function in regulating diverse physiological processes in swamp eel.

Two new species of the harvestman genus Sabacon Simon, 1879 are recorded from Yunnan Province, southern China, Sabacon monacanthus sp. n. (male and female; relationships unclear) and S. suzukii sp. n. (male and female; closely related to S. kangding Martens, 2015 from Sichuan Province, southwestern China).

Blood vessels, together with the heart, have a fundamental role in supporting the metabolic demands of tissues not only during development but also in adults. New blood vessels are frequently generated through angiogenesis when new vessels emerge from pre-existing ones (Fig. 2.1a). Initially, endothelial cells (ECs) lining an existing vessel are selected to become tip cells to spearhead the formation of new vascular sprouts. New sprouts grow through EC proliferation and the polarized collective migration of both tip and trailing stalk cells into the avascular tissue. In order to generate a network of interconnecting vessel segments, tip cells anastomose with neighboring tip cells to establish new vascular loops. Importantly, vascular sprouts develop into tubes through which oxygen, metabolites, cells, and waste products can circulate around the body. Finally, the tubular network of blood vessels are either maintained or, depending on the tissue requirements in which the vessels pervade, remodeled through pruning into a more refined vascular network that carries blood flow optimally to tissues (Fig. 2.1b).

Colon cancer is one of the most life-threatening malignancies worldwide. Long noncoding RNA (lncRNA) HOX transcript antisense RNA (HOTAIR) is a cancer-associated biomarker involved in the metastasis and prognosis of several cancers. However, whether and how HOTAIR affects colon cancer progression is still unclear. Consequently, we used RNA interference to knock down HOTAIR to explore its effects on human colon cancer cells. The dual luciferase reporter gene assay was initially used for testify the regulating relationship between lncRNA HOTAIR and insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2). We determined the expressions of HOTAIR, IGF2BP2, E-cadherin, and vimentin. Meanwhile, cell growth, cycle and apoptosis, migration, and invasion were assayed. LoVo cells were transplanted into nude mice, and tumor formation and microvessel density were evaluated. LncRNA HOTAIR positively regulated IGF2BP2. Besides, the expressions of HOTAIR and E-cadherin and the apoptosis were increased, while the expressions of IGF2BP2 and vimentin, the growth, invasion and migration of LoVo cells, the average tumor weight, and microvessel density value were decreased. Of importance, overexpressed IGF2BP2 could reverse the above impacts. Taken together, the current study indicates that silencing of HOTAIR could inhibit the invasion, proliferation, and migration, and promote apoptosis of colon cancer LoVo cells through suppressing IGF2BP2 and the epithelial-mesenchymal transition.

The endo-inulinase genes from Aspergillus niger (A. niger) TCCC41064 and Penicillium sp. TN-88 were inserted into the expression vector pPIC9 K. The recombinant plasmids were subsequently transformed into P. pastoris GS115 by electroporation, respectively. The two recombinants (P. pastoris GS115/pPIC9 K-inuA, GS115/pPIC9 K-inuC) were purified and characterized. The molecular weight of the purified inuA and inuC were both about 66.2 kDa. The specific activity of transformant endo-inulinase inuA and inuC were 13.5 and 69.3 U/mL. The optimal pH and temperature were 5.0, 60 °C for inuA, and 4.0, 50 °C, respectively. InuA, 83.2% of the residual enzymatic activity at 55 °C for 9 h. Meanwhile, inuC retained 75.4% at 40 °C for 9 h. The purified inuC converts inulin into trisaccharide (74.8%) and for the inuA was 36.7%. The researches on inulinase will provide a better engineering strain for designing the industrial biocatalyst.