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Platelet over-activation is one of the important mechanisms of atherosclerosis （AS） in diabetes mellitus. Dietary nutrient intervention plays an important role in the primary prevention and treatment of diabetes mellitus combined with AS. Studies have shown that the antiplatelet factor in tomato extract can inhibit platelet aggregation and activation in different environments. Its bioactive ingredient are nucleosides and derivatives and phenolic compounds， which retard the pathological progress of diabetes mellitus complicated with AS. This review aims to explore the mechanism of the antiplatelet effect of tomato extract， and to provide a theoretical basis for the prevention and treatment of diabetes mellitus complicated with AS.

Objective To explore the role and underlying mechanism of C1q/TNF-related protein 6 (CTRP6) in the development process of liver disease. Methods The histopathological examination and biochemical indexes of injured livers from wild-type (CTRP6+/+) and knockout (CTRP6-/-) mice induced by intragastric administration of CCl4 were analyzed. RT-qPCR was used to detect the expression of the fibrosis marker genes (Acta2, Col1al, Mmp2 and TGF-β) and cell proliferation marker genes (Cyclin D, Cyclin E and CDK6). Results Compared with CTRP6+/+ mice, the hepatocytes in CTRP6-/- mice showed mild necrosis, looseness and swelling. The high SOD activity and low MDA level were also observed in CTRP6-/- mice liver. Meanwhile, the expression of fibrosis and cell proliferation markers in CTRP6-/- mice were significantly lower than those in CTRP6+/+ mice. Conclusions CTRP6 gene is closely related to liver disease development, and the liver fibrosis progression induced by CCl4 is alleviated by its deletion. The mechanism may be explained as that CTRP6 knockout affects the proliferative capacity of hepatocytes.

A new atisane-type diterpene, ent-16α-hydroxy-atisane-3,4-lactone (4) and three known diterpenes, ent-16α-hydroxy-atisane-3-one (1), ent-atisane-3β,16α-diol (2), ent-3,4-seco-16α-hydroxyatis- 4(19)-en -3-oic acid (3) were isolated from the bark of the mangrove plant Excoecaria agallocha. Their structures and relative stereochemistry were elucidated by means of extensive NMR and MS analysis. Compound 3 exhibited significant anti-microfouling activity against the adherence of Pseudomonas pseudoalcaligenes, with an EC50 value of 0.54 ± 0.01 ppm.

Abstract Inspired by the concept of “natural camouflage,” biomimetic drug delivery systems have emerged to address the limitations of traditional synthetic nanocarriers, such as poor targeting, susceptibility to identification and clearance, inadequate biocompatibility, low permeability, and systemic toxicity. Biomimetic nanocarriers retain the proteins, nucleic acids, and other components of the parent cells. They not only facilitate drug delivery but also serve as communication media to inhibit tumor cells. This paper delves into the communication mechanisms between various cell-derived biomimetic nanocarriers, tumor cells, and the tumor microenvironment, as well as their applications in drug delivery. In addition, the additional communication capabilities conferred on the modified biomimetic nanocarriers, such as targeting and environmental responsiveness, are outlined. Finally, we propose future development directions for biomimetic nanocarriers, hoping to inspire researchers in their design efforts and ultimately achieve clinical translation. Graphical Abstract

Abstract Talin2 is localized to large focal adhesions and is indispensable for traction force generation, invadopodium formation, cell invasion as well as metastasis. Talin2 has a higher affinity toward β-integrin tails than talin1. Moreover, disruption of the talin2-β-integrin interaction inhibits traction force generation, invadopodium formation and cell invasion, indicating that a strong talin2-β-integrin interaction is required for talin2 to fulfill these functions. Nevertheless, the role of talin2 in mediation of these processes remains unknown. Here we show that talin2 binds to the N-terminus of non-muscle myosin IIA (NMIIA) through its F3 subdomain. Moreover, talin2 co-localizes with NMIIA at cell edges as well as at some cytoplasmic spots. Talin2 also co-localizes with cortactin, an invadopodium marker. Furthermore, overexpression of NMIIA promoted the talin2 head binding to the β1-integrin tail, whereas knockdown of NMIIA reduced fibronectin and matrix metalloproteinase secretion as well as inhibited cell attachment on fibronectin-coated substrates. These results suggest that talin2 binds to NMIIA to control the secretion of extracellular matrix proteins and this interaction modulates cell adhesion.

Objective To evaluate the efficacy and safety of early use of new oral anticoagulants (≤3 days) in patients with acute ischemic stroke(AIS) due to atrial fibrillation (AF).Method After systematic retrieval of CNKI, WanFang Data, VIP, PubMed, The Cochrane Library and Embase database, clinical studies on early anticoagulation therapy for AF-related AIS patients were screened. The search was conducted from the time of database construction to November 4, 2022, while the references were searched manually. Literature screening, data extraction and quality assessment were performed independently by 2 investigators. Meta-analysis was performed using RevMan 5.4 software, and the combined dichotomous effect sizes were expressed as relative risk (RR) and 95% confidence interval (CI).Result A total of 2 randomized controlled trials and 5 non-randomized controlled trials including 2 226 patients were included, and the analysis suggested that early anticoagulation treatment (≤3 days) was similar to delayed anticoagulation (> 3 days) in terms of total ischemic events [RR=1.04, 95%CI(0.67, 1.61), P=0.88], recurrent ischemic strokes or transient ischemic attack [RR=0.88, 95%CI(0.58, 1.34), P=0.56], major bleeding [RR=1.24, 95%CI(0.59, 2.62), P=0.56], and symptomatic intracerebral haemorrhage [RR=0.98, 95%CI(0.44, 2.17), P=0.96]. In terms of total cerebral hemorrhages, the incidence was lower than in the delayed anticoagulation group.Conclusion Anticoagulation started early after the occurrence of atrial fibrillation related ischemic stroke is similar to delayed anticoagulation in terms of effectiveness, and does not increase the risk of bleeding in patients.

【Objective】 To explore the variation characteristics of kiwifruit sap flow and its response to environmental factors. 【Method】 The thermal diffusion pin-type sap flow meter was used to monitor the sap flow law of kiwifruit trees in Guanzhong Plain, and the relationship between the sap flow of kiwifruit tree and environmental factors was analyzed. 【Result】 The daily-scale sap flow of kiwifruit tree showed a significant single-peak change during the whole growth period. The maximum value was 11.00 cm/h, which appeared in the fruit expansion period, and the minimum value was 0.18 cm/h, which appeared in the leaf expansion period. The peak value of hourly-scale sap flow rate in each growth period was: fruit expansion period > fruit maturity period > flowering and fruit setting period>shoot expansion period. The peak value of sap flow rate under different weather conditions was: sunny day>cloudy day > rainy day. The daily average sap flow rate of fruit tree was significantly positively correlated with air temperature, water vapor pressure deficit and net solar radiation, and the correlation coefficients were 0.842, 0.749 and 0.713, respectively. The results of path analysis showed that the net solar radiation had the greatest direct effect on the sap flow of kiwifruit tree, and the soil volumetric water content mainly affected the plant sap flow through the combined effect of net solar radiation, air temperature and soil temperature. There is an obvious time lag effect between meteorological factors and liquid flow rate. The time lag between liquid flow rate and solar net radiation is -30 min, the time lag between liquid flow rate and humidity is 60 min, and the time lag between liquid flow rate and air temperature and water vapor pressure deficit is 90 min. 【Conclusion】 The transpiration and water consumption of kiwifruit tree during fruit expansion period were the largest, so attention should be paid to the soil water status of kiwifruit tree during this period. In sunny weather, the sap flow of kiwifruit tree is the largest from 12:00 to 16:00, and the evaporation is also large, so irrigation should be avoided in this time to reduce the irrigation loss caused by transpiration.

Thermal barrier coatings, which are widely used in aero-engines and gas turbines, can reduce the surface temperature and improve the effective service time of superalloy due to low thermal conductivity and good temperature resistance. The performance and lifetime of the thermal barrier coating are directly affected by the material and structure of the ceramic top coating. The method of adjusting the microstructure of ceramic coating with controlled raw material powder can reduce the strain-stress mismatch in the coating, and has the advantages of flexible operation, remarkable effect and wide control range. In order to solve problems of low strain tolerance and insufficient thermal shock resistance in traditional coatings, our team successfully has prepared ceramic microspheres with hierarchy pore structure by using electrostatic spraying combined with phase inversion theory（ESP）. Compared with traditional hollow microspheres, the ESP microstructure of nano-pores and finger-like pores lead to high sintering resistance, low thermal conductivity, high specific strength and >95% thermal reflectivity. With the retention of the hierarchy structure, the coating has good fracture toughness and strain tolerance, where the thermal cycle life of the coating is increased by more than ~2 times. ESP technology provides a fast feedback control method for new thermal barrier coating materials from material design to engineering application, such as rare earth zirconate, rare earth tantalate and rare earth doped YSZ high entropy system. With the in-depth study of material mechanics, optics, thermodynamics and the precise control of internal topological structure, the hierarchy pore structure will be more widely used in aerospace, military defense, fluorescence temperature measurement and other fields in the future.

Abstract The landscape of current cancer immunotherapy is dominated by antibodies targeting PD-1/PD-L1 and CTLA-4 that have transformed cancer therapy, yet their efficacy is limited by primary and acquired resistance. The blockade of additional immune checkpoints, especially TIGIT and LAG-3, has been extensively explored, but so far only a LAG-3 antibody has been approved for combination with nivolumab to treat unresectable or metastatic melanoma. Here we report the development of a PDL1 × TIGIT bi-specific antibody (bsAb) GB265, a PDL1 × LAG3 bsAb GB266, and a PDL1 × TIGIT × LAG3 tri-specific antibody (tsAb) GB266T, all with intact Fc function. In in vitro cell-based assays, these antibodies promote greater T cell expansion and tumor cell killing than benchmark antibodies and antibody combinations in an Fc-dependent manner, likely by facilitating T cell interactions (bridging) with cancer cells and monocytes, in addition to blocking immune checkpoints. In animal models, GB265 and GB266T antibodies outperformed benchmarks in tumor suppression. This study demonstrates the potential of a new generation of multispecific checkpoint inhibitors to overcome resistance to current monospecific checkpoint antibodies or their combinations for the treatment of human cancers.

【Objective】 Variation in crop yield is the consequence of many natural and anthropogenic factors, and disentangling their impacts is important for improving agricultural management but difficult. The purpose of this paper is to propose and compared different methods to isolate the impacts of meteorological change on crop yield based on long time series of maize yield in Yellow River-water irrigated region in Ninxia province of China. 【Method】 The analysis is based on maize yield measured from1988 to 2019 in 6 counties located in the Yellow River-watered irrigation areas. We compared three methods for the separation: five -year moving average method, quadratic exponential smoothing method, and five-point quadratic smoothing method. The consistent correlation coefficient, trend coincidence conformity analysis method, consistency of climate change characteristics, which lead to the same rise-fall in meteorological yield, were used as the evaluation criteria. Their applicability and rationality were compared and analyzed. All methods were calibrated based on the relationship between meteorological factors and maize yield. 【Result】 All methods can fit the yield trend well. Compared with the average yield trend, the consistency correlation coefficients of all three methods were >0.5, suggesting that there was no significant difference between these methods for fitting the yield trend. The advantage of the quadratic exponential smoothing method and the five-point quadratic smoothing method is that they accurately describe the change in the yield as affected by national productivity and national policy. The change in the yield due to meteorological factors estimated by the five-point quadratic smoothing method described the effect of inter-annual meteorological factors better, and its associated meteorological yield model is able to describe the relationship between the meteorological factors and the maize yield. 【Conclusion】 Comprehensive analysis showed that the five-point quadratic smoothing method modeled the yield change due to meteorological factors better than the other two methods.

Backgrounds: The incidence of melanosis coli (MC) has gradually increased annually, attracting significant attention and efforts into this field. A potential risk for MC is the long-term use of anthraquinone laxatives in patients with constipation. Most traditional cathartic drugs are made from herbs containing anthraquinone compounds. This review aims to provide guidance for the application of traditional Chinese herbs containing anthraquinones for physicians and researchers.Materials and methods: We reviewed risk factors and pathogenesis of MC, and natural anthraquinones isolated from TCM herbs. We searched Pubmed and CNKI databases for literature related to MC with keywords such as“traditional Chinese medicine”, “Chinese herbs”, “anthraquinones”, and “melanosis coli”. The literature is current to January 2023 when the searches were last completed. After the literature retrieval, the TCM herbs containing anthraquinones (including component identification and anthraquinone content determination) applied in clinical were selected. According to the collected evidence, we provide a list of herbs containing anthraquinones that could cause MC.Results: We identified 20 herbs belonging to 7 families represented by Polygonaceae, Fabaceae, Rhamnaceae, and Rubiaceae, which may play a role in the pathogenesis of MC. Among these, the herbs most commonly used include Dahuang (Rhei Radix et Rhizome), Heshouwu (Radix Polygoni Multiflori), Huzhang (Rhizoma Polygoni Cuspidati), Juemingzi (Semen Cassiae), Luhui (Aloe) and Qiancao (Rubiae Radix et Rhizoma).Conclusion: Due to a lack of awareness of the chemical composition of TCM herbs, many patients with constipation and even some TCM physicians take cathartic herbal remedies containing abundant anthraquinones to relieve defecation disturbances, resulting in long-term dependence on these herbs, which is potentially associated with most cases of MC. When such treatments are prescribed, TCM physicians should avoid long-term use in large doses to reduce their harm on colonic health. Individuals who take healthcare products containing these herbs should also be under the supervision of a doctor.

【Objective】 Deficit irrigation is a technology used widely in most arid and semi-arid regions to increase water use efficiency, but how to optimize the timing of and duration of the deficit remains elusive. The objective of this paper is to fill this gap for summer maize grown in the north of China. 【Method】 The field experiment compared three deficit irrigations by controlling the irrigation water amount at 100% (sufficient), 80% (moderate deficit) and 60% (sever deficit) of the potential evapotranspiration measured from a lysimeter (ETa). Each treatment irrigated the crop at emergence-jointing stage, jointing-tasseling stage, and tasseling-filling stage, respectively. Overall, there were nine treatments: moderate deficit irrigation at the jointing-filling stage (T1), severe deficit irrigation at the jointing-filling stage (T2), moderate deficit irrigation at the emergence-jointing and tasseling-filling stage (T3), moderate deficit irrigation at the emergence-tasseling stage and severe deficit irrigation at the tasseling-filling stage (T4), moderate deficit irrigation at the emergence-jointing stage and severe deficit irrigation at the jointing-tasseling stage (T5), severe deficit irrigation at the emergence-jointing and tasseling-filling stage (T6), severe deficit irrigation at the emergence-jointing stage and moderate deficit irrigation at jointing-tasseling stage (T7), severe deficit irrigation at the emergence-tasseling stage (T8), and moderate deficit at the tasseling-filling stage (T8). Sufficient irrigation in all stages was taken as the control (CK). In each treatment we measured the impact of rewatering on evapotranspiration, grain yield and water use efficiency of the crop. 【Result】 Compared with CK, deficit irrigation led to a decrease in evapotranspiration regardless of the stage it was imposed. Rewatering in T3 increased the evapotranspiration to a level comparable to that in CK. T5, T6 and T7 resulted in a permanent reduction in evapotranspiration even after rewatering. To avoid grain yield loss, the soil water content during the tasseling-filling stage should be maintained above 75% of the field capacity. The critical soil water was 50% of the field capacity, below which the water taken up by the crop was unable to sustain its growth. Deficit irrigation reduced the fruit yield by 5.85%～32.25%, compared to the CK, with T3 reducing the least (5.85%). T3 was thus most water-use efficient (2.81 kg/m3), increasing water use efficiency by 4.07% compared to CK. 【Conclusion】 Moderate water deficit at the seedling stage and full irrigation at the jointing-tasseling stage were optimal to improve water use efficiency without significantly reducing grain yield of the maize.

Abstract Cells adhere to the surrounding tissue and probe its mechanical properties by forming cell-matrix adhesions. Talin is a critical adhesion protein and participates in the transmission of mechanical signals between extracellular matrix and cell cytoskeleton. Force induced unfolding of talin rod subdomains has been proposed to act as a cellular mechanosensor, but so far evidence linking their mechanical stability and cellular response has been lacking. Here, by utilizing computationally designed mutations, we demonstrate that stepwise destabilization of the talin rod R3 subdomain decreases cellular traction force generation, which affects talin and vinculin dynamics in cell-matrix adhesions and results in the formation of talin-rich but unstable adhesions. We observed a connection between talin stability and the rate of cell migration and also found that talin destabilization affects the usage of different integrin subtypes and sensing of extracellular matrix proteins. Experiments with truncated forms of talin confirm the mechanosensory role of the talin R3 subdomain and exclude the possibility that the observed effects are caused by the release of talin head-rod autoinhibition. In conclusion, this study provides evidence into how the controlled talin rod domain unfolding acts as a key regulator of adhesion structure and function and consequently controls central cellular processes such as cell migration and substrate sensing.

Increased expression of NOX4 protein is associated with cancer progression and metastasis but the role of NOX4 in cell proliferation and invasion is not fully understood. We generated NOX4 knockout HeLa cell lines using the CRISPR-Cas9 gene editing system to explore the cellular functions of NOX4. After transfection of CRISPR-Cas9 construct, we performed T7 endonuclease 1 assays and DNA sequencing to generate and identify insertion and deletion of the NOX4 locus. We confirmed the knockout of NOX4 by Western blotting. NOX4 knockout cell lines showed reduced cell proliferation with an increase of sub-G1 cell population and the decrease of S/G2/M population. Moreover, NOX4 deficiency resulted in a dramatic decrease in invadopodium formation and the invasive activity. In addition, NOX4 deficiency also caused a decrease in focal adhesions and cell migration in HeLa cells. These results suggest that NOX4 is required for both efficient proliferation and invasion of HeLa cells.

Precision medicine is a rapidly growing area of modern medical science and open source machine-learning codes promise to be a critical component for the successful development of standardized and automated analysis of patient data. One important goal of precision cancer medicine is the accurate prediction of optimal drug therapies from the genomic profiles of individual patient tumors. We introduce here an open source software platform that employs a highly versatile support vector machine (SVM) algorithm combined with a standard recursive feature elimination (RFE) approach to predict personalized drug responses from gene expression profiles. Drug specific models were built using gene expression and drug response data from the National Cancer Institute panel of 60 human cancer cell lines (NCI-60). The models are highly accurate in predicting the drug responsiveness of a variety of cancer cell lines including those comprising the recent NCI-DREAM Challenge. We demonstrate that predictive accuracy is optimized when the learning dataset utilizes all probe-set expression values from a diversity of cancer cell types without pre-filtering for genes generally considered to be "drivers" of cancer onset/progression. Application of our models to publically available ovarian cancer (OC) patient gene expression datasets generated predictions consistent with observed responses previously reported in the literature. By making our algorithm "open source", we hope to facilitate its testing in a variety of cancer types and contexts leading to community-driven improvements and refinements in subsequent applications.

Detection of protein-protein interactions in cells is crucial for understanding the biological functions of proteins, including their roles in signal transduction. However, current methods require specific antibodies both for immunoprecipitation and detection, making them expensive and sometimes unreliable. Here we describe protocols for protein-protein interaction assays that use nonimmune IgG-conjugated Sepharose to precipitate the IgG binding domain (ZZ) fused to the bait protein; the interaction partner is fused to Avitag and biotinylated by BirA so that it can be detected by a one-step blot with Dylight 680 streptavidin to detect the Avitag fusion protein. Since this method does not require specific antibodies and is inexpensive, sensitive, and reliable, it should be useful for detecting protein-protein interactions in cells.

Yaks (Bos grunniens) are endemic species that can adapt well to thin air, cold temperatures, and high altitude. These species can survive in harsh plateau environments and are major source of animal production for local residents, being an important breed in the Qinghai-Tibet Plateau. However, compared with ordinary cattle that live in the plains, yaks generally have lower fertility. Investigating the basic physiological molecular features of yak ovary and identifying the biological events underlying the differences between the ovaries of yak and plain cattle is necessary to understand the specificity of yak reproduction. Therefore, RNA-seq technology was applied to analyze transcriptome data comparatively between the yak and plain cattle estrous ovaries.After deep sequencing, 3,653,032 clean reads with a total of 4,828,772,880 base pairs were obtained from yak ovary library. Alignment analysis showed that 16992 yak genes mapped to the yak genome, among which, 12,731 and 14,631 genes were assigned to Gene Ontology (GO) categories and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Furthermore, comparison of yak and cattle ovary transcriptome data revealed that 1307 genes were significantly and differentially expressed between the two libraries, wherein 661 genes were upregulated and 646 genes were downregulated in yak ovary. Functional analysis showed that the differentially expressed genes were involved in various Gene Ontology (GO) categories and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. GO annotations indicated that the genes related to "cell adhesion," "hormonal" biological processes, and "calcium ion binding," "cation transmembrane transport" molecular events were significantly active. KEGG pathway analysis showed that the "complement and coagulation cascade" pathway was the most enriched in yak ovary transcriptome data, followed by the "cytochrome P450" related and "ECM-receptor interaction" pathways. Moreover, several novel pathways, such as "circadian rhythm," were significantly enriched despite having no evident associations with the reproductive function.Our findings provide a molecular resource for further investigation of the general molecular mechanism of yak ovary and offer new insights to understand comprehensively the specificity of yak reproduction.

The issue of how contractility and adhesion are related to cell shape and migration pattern remains largely unresolved. In this paper we report that Gleevec (Imatinib), an Abl family kinase inhibitor, produces a profound change in the shape and migration of rat bladder tumor cells (NBTII) plated on collagen-coated substrates. Cells treated with Gleevec adopt a highly spread D-shape and migrate more rapidly with greater persistence. Accompanying this more spread state is an increase in integrin-mediated adhesion coupled with increases in the size and number of discrete adhesions. In addition, both total internal reflection fluorescence microscopy (TIRFM) and interference reflection microscopy (IRM) revealed a band of small punctate adhesions with rapid turnover near the cell leading margin. These changes led to an increase in global cell-substrate adhesion strength, as assessed by laminar flow experiments. Gleevec-treated cells have greater RhoA activity which, via myosin activation, led to an increase in the magnitude of total traction force applied to the substrate. These chemical and physical alterations upon Gleevec treatment produce the dramatic change in morphology and migration that is observed.

Focal adhesion assembly and disassembly are essential for cell migration and cancer invasion, but the detailed molecular mechanisms regulating these processes remain to be elucidated. Phosphatidylinositol phosphate kinase type Iγ (PIPKIγ) binds talin and is required for focal adhesion formation in EGF-stimulated cells, but its role in regulating focal adhesion dynamics and cancer invasion is poorly understood. We show here that overexpression of PIPKIγ promoted focal adhesion formation, whereas cells expressing either PIPKIγ(K188,200R) or PIPKIγ(D316K), two kinase-dead mutants, had much fewer focal adhesions than those expressing WT PIPKIγ in CHO-K1 cells and HCT116 colon cancer cells. Furthermore, overexpression of PIPKIγ, but not PIPKIγ(K188,200R), resulted in an increase in both focal adhesion assembly and disassembly rates. Depletion of PIPKIγ by using shRNA strongly inhibited formation of focal adhesions in HCT116 cells. Overexpression of PIPKIγ(K188,200R) or depletion of PIPKIγ reduced the strength of HCT116 cell adhesion to fibronection and inhibited the invasive capacities of HCT116 cells. PIPKIγ depletion reduced PIP₂ levels to ∼40% of control and PIP₃ to undetectable levels, and inhibited vinculin localizing to focal adhesions. Taken together, PIPKIγ positively regulates focal adhesion dynamics and cancer invasion, most probably through PIP₂-mediated vinculin activation.

Abstract Transcriptomic analysis has been widely used in comparative experiments to uncover biological mechanisms in various species. However, a simple tool is still lacking to optimize and integrate the features from multiple R packages. In this study, we developed TOmicsVis (Transcriptomics Visualization) (CRAN: https://cran.r-project.org/package=TOmicsVis, v2.0.0), an R package that provides a comprehensive solution for transcriptomics analysis and visualization. It utilizes 46 R packages to design 40 suitable functions for the streamlined analysis of multigroup transcriptomic projects, which covers six main categories: Sample Statistics, Traits Analysis, Differential Expression, Advanced Analysis, GO and KEGG Enrichment, and Table Operation. TOmicsVis can be performed either locally or online (https://shiny.hiplot.cn/tomicsvis-shiny/), which provides significant convenience for researchers without coding training. These user‐friendly visualization functions and built‐in analysis capabilities enable researchers to monitor experimental data dynamics promptly and explore transcriptomics data quickly.

Endocannabinoids play important roles in the functioning of various physiological systems in humans and non-mammalian animals, including invertebrates. However, information concerning their roles in physiological functions in members of the phylum Mollusca is scarce. Here the hypothesis that the endocannabinoids are involved in mediating settlement of marine invertebrates was tested. Two endocannabinoids [N-arachidonoyl ethanolamide (AEA) and 2-arachidonoyl glycerol (2-AG)], and two endocannabinoid-like lipids [N-Oleoylethanolamide (OEA) and N-Palmitoylethanolamide (PEA)] were detected in the green mussel Perna viridis. In particular, 2-AG was present at significantly higher levels in unattached P. viridis compared with attached mussels. The in vivo level of 2-AG was inversely correlated with the attachment activity of P. viridis. Furthermore, exposure to synthetic 2-AG inhibited attachment of P. viridis in a reversible manner. Transcriptomic analysis suggested that up-regulation of 2-AG synthase (Phospholipase C-β, PLC-β) and down-regulation of its degrading enzyme (Monoacylglycerol lipase, MAGL) resulted in higher levels of 2-AG in unattached mussels. A putative mechanism for the negative regulation of mussel attachment by 2-AG is proposed that involves a Ca2+- Nitric oxide (NO)- cyclic guanosine monophosphate (cGMP) pathway. This study broadens our understanding of the evolution and roles of the endocannabinoid system in animals, and reveals an endogenous regulatory cue for mussel attachment.

The mangrove endophytic fungus Aspergillus terreus (No. GX7-3B) was cultivated in potato dextrose liquid medium, and one rare thiophene compound (1), together with anhydrojavanicin (2), 8-O-methylbostrycoidin (3), 8-O-methyljavanicin (4), botryosphaerone D (5), 6-ethyl-5-hydroxy-3,7-dimethoxynaphthoquinone (6), 3β,5α-dihydroxy-(22E,24R)-ergosta-7,22-dien-6-one (7), 3β,5α,14α-trihydroxy-(22E,24R)-ergosta-7, 22-dien-6-one (8), NGA0187 (9) and beauvericin (10), were isolated. Their structures were elucidated by analysis of spectroscopic data. This is the first report of a natural origin for compound 6. Moreover, compounds 3, 4, 5, 7, 8 and 10 were obtained from marine microorganism for the first time. In the bioactive assays in vitro, compounds 2, 3, 9 and 10 displayed remarkable inhibiting actions against α-acetylcholinesterase (AChE) with IC50 values 2.01, 6.71, 1.89, and 3.09 μM, respectively. Furthermore, in the cytotoxicity assays, compounds 7 and 10 exhibited strong or moderate cytotoxic activities against MCF-7, A549, Hela and KB cell lines with IC50 values 4.98 and 2.02 (MCF-7), 1.95 and 0.82 (A549), 0.68 and 1.14 (Hela), and 1.50 and 1.10 μM (KB), respectively; compound 8 had weak inhibitory activities against these tumor cell lines; compounds 1, 2, 3, 4, 5, 6 and 9 exhibited no inhibitory activities against them.

Three new bianthraquinone derivatives, alterporriol K (1), L (2) and M (3), along with six known compounds were obtained from extracts of the endophytic fungus Alternaria sp. ZJ9-6B, isolated from the mangrove Aegiceras corniculatum collected in the South China Sea. Their structures were elucidated by one- and two-dimensional NMR spectroscopy, MS data analysis and circular dichroism measurements. Compounds 1, 2 and 3 were first isolated alterporriols with a C-2–C-2′ linkage. The crystallographic data of tetrahydroaltersolanol B (7) was reported for the first time. In the primary bioassays, alterporriol K and L exhibited moderate cytotoxic activity towards MDA-MB-435 and MCF-7 cells with IC50 values ranging from 13.1 to 29.1 µM.

The small abalone Haliotis diversicolor is an economically important mollusk that is widely cultivated in Southern China. Gonad precocity may affect the aquaculture of small abalone. Polyamines, which are small cationic molecules essential for cellular proliferation, may affect gonadal development. Ornithine decarboxylase (ODC) and antizyme (AZ) are essential elements of a feedback circuit that regulates cellular polyamines. This paper presents the molecular cloning and characterization of AZ from small abalone. Sequence analysis showed that the cDNA sequence of H. diversicolor AZ (HdiODCAZ) consisted of two overlapping open reading frames (ORFs) and conformed to the +1 frameshift property of the frame. Thin Layer chromatography (TLC) analysis suggested that the expressed protein encoded by +1 ORF2 was the functional AZ that targets ODC to 26S proteasome degradation. The result demonstrated that the expression level of AZ was higher than that of ODC in the ovary of small abalone. In addition, the expression profiles of ODC and AZ at the different development stages of the ovary indicated that these two genes might be involved in the gonadal development of small abalone.

BACKGROUND: The abalone Haliotis diversicolor is a good model for study of the settlement and metamorphosis, which are widespread marine ecological phenomena. However, information on the global gene backgrounds and gene expression profiles for the early development of abalones is lacking. METHODOLOGY/PRINCIPAL FINDINGS: In this study, eight non-normalized and multiplex barcode-labeled transcriptomes were sequenced using a 454 GS system to cover the early developmental stages of the abalone H. diversicolor. The assembly generated 35,415 unigenes, of which 7,566 were assigned GO terms. A global gene expression profile containing 636 scaffolds/contigs was constructed and was proven reliable using qPCR evaluation. It indicated that there may be existing dramatic phase transitions. Bioprocesses were proposed, including the 'lock system' in mature eggs, the collagen shells of the trochophore larvae and the development of chambered extracellular matrix (ECM) structures within the earliest postlarvae. CONCLUSION: This study globally details the first 454 sequencing data for larval stages of H. diversicolor. A basic analysis of the larval transcriptomes and cluster of the gene expression profile indicates that each stage possesses a batch of specific genes that are indispensable during embryonic development, especially during the two-cell, trochophore and early postlarval stages. These data will provide a fundamental resource for future physiological works on abalones, revealing the mechanisms of settlement and metamorphosis at the molecular level.

Genetic data were used to measure the phylogeographic distribution of the limpet, Cellana toreuma along the China coast in order to acsertain impacts of historic events, ocean currents and especially freshwater discharge from the Yangtze River on the connectivity of intertidal species with limited larval dispersal capability.Genetic variation in 15 populations of C. toreuma (n = 418), ranging from the Yellow Sea (YS), East China Sea (ECS) and South China Sea (SCS), were determined from partial mitochondrial cytochrome c oxidase subunit I gene. Genetic diversity and divergence based on haplotype frequencies were analyzed using CONTRIB, and AMOVA was used to examine genetic population structure. Historic demographic expansions were evaluated from both neutrality tests and mismatch distribution tests. Among the 30 haplotypes identified, a dominant haplotype No. 1 (H1) existed in all the populations, and a relatively abundant private haplotype (H2) in YS. Pairwise F(ST) values between YS and the other two groups were relatively high and the percentage of variation among groups was 10.9%.The high nucleotide and gene diversity in the YS, with large pairwise genetic distances and relatively high percentages of variation among groups, suggests that this group was relatively isolated from ECS and SCS. This is likely driven by historic events, ocean currents, and demographic expansion. We propose that freshwater discharge from the Yangtze River, which may act as physical barrier limiting the southward dispersal of larvae from northern populations, is especially important in determining the separation of the YS group from the rest of the Chinese populations of C. toreuma.