Changes in intracellular and extracellular proteins after ERGIC3 knockdown in lung cancer:Proteins interacting with ERGIC3,HORN,and FLNA

Objective:Endoplasmic reticulum-Golgi intermediate compartment 3(ERGIC3)promotes cell proliferation and metastasis in lung cancer,but its molecular mechanism is unclear.Methods:The GLC-82 cells were randomly divided into the ERGIC3i group and the negative control group.The cells were transfected with ERGIC3 siRNA or control siRNA in the groups,respectively.The ERGIC3-interacting proteins expressed in cells or extracellularly were isolated by the immunoprecipitation method and detected by isobaric tags for relative and absolute quantitation and liquid chromatography-tandem mass spectrometry.The differentially expressed proteins were determined by bioinformatic methods.Results:After ERGIC3 knockdown,88 extracellular differentially expressed proteins,41 upregulated and 47 down-regulated,were detected in the supernatant of cultured cells.Among 52 intracellular differentially expressed proteins,33 were up-regulated and 19 down-regulated.Cluster analysis showed that the extracellular differential proteins are mainly involved in Ca2+binding and transport and I-kappa B kinase/NF-kappa B signal transduction.The upregulated proteins are mainly involved in the biological process of H3-K27 and H3-K4 methylation in cells.Co-immunoprecipitation assay showed that proteins interacting with ERGIC3 were rich in cytoskeleton construction and RHO GTPases activated p21-activated kinases.The intersection of these two research methods shows that ERGIC3 interacts with HORN and filamin A(FLNA).Conclusion:Proteomic analyses reveal that ERGIC3 acts as a vesicle transmembrane protein on the distribution of various extracellular and intracellular proteins and regulates the extracellular and intracellular biological processes by specifically binding hornin(HORN)and FLNA proteins.These findings maybe provide new methods and ideas for ERGIC3 as a therapeutic target for lung cancer.

Seasonal dynamic variation of pollination network is associated with the number of species in flower in an oceanic island community

Aims Plant-pollinator interaction networks are dynamic entities,and seasonal variation in plant phenology can reshape their structure on both short and long timescales.However,such seasonal dynamics are rarely considered,especially for oceanic island pollination networks.Here,we assess changes in the temporal dynamics of plant-pollinator interactions in response to seasonal variation in floral resource richness in oceanic island communities.Methods We evaluated seasonal variations of pollination networks in the Yongxing Island community.Four temporal qualitative pollination networks were analyzed using plant-pollinator interaction data of the four seasons.We collected data on plant-pollinator interactions during two consecutive months in each of the four seasons.Four network-level indices were calculated to characterize the overall structure of the networks.Statistical analyses of community dissimilarity were used to compare this community across four seasons to explore the underlying factors driving these patterns.We also evaluated the temporal variation in two species-level indices of plant and pollinator functional groups.Important Findings Both network-level specialization and modularity showed a significantly opposite trend compared with plant species richness across four seasons.Increased numbers of plant species might promote greater competition among pollinators,leading to increased niche overlap and causing decreased specialization and modularity and vice versa.Further analyses suggested that the season-to-season turnover of interactions was dominated by interaction rewiring.Thus,the seasonal changes in niche overlap among pollinators lead to interaction rewiring,which drives interaction turnover in this community.Hawkmoths had higher values of specialization and Apidae had higher values of species strength compared with other pollinator functional groups.These findings should be considered when exploring plant-pollinator interactions in ecosystems of isolated oceanic islands and in other ecosystems.

Characterization of chlorine dioxide as disinfectant for the removal of low concentration microcystins

Microcystins, which represents one kind of cancerogenic organic compounds, is abundant in eutro- phication water. The effects of reaction factors on chlorine dioxide （C102） for removal of low-concentration Micro- cystin-LR, Microcystin-RR, and Microcystin-YR in water as well as the reaction mechanisms was investigated by using enzyme-linked immunosorbent assay （ELISA） kit and gas chromatography-mass spectrometry （GC-MS）. The results showed that MC-LR, MC-RR, and MC-YR could be efficiently decomposed by C102. The degradation efficiency was shown positively correlated to the concen- tration of C102 and reaction time; while the effect of reaction temperature and pH is slight. The kinetic constants and activation energies of the reaction ofMC-LR, MC-RR, and MC-YR with C102 are determined as 459.89, 583.15, 488.43 L.（mol.min）I and 64.78, 53.01, 59.15kJ.mok1, respectively. As indicated by high performance liquid chromatography mass spectrometer （HPLC-MS） analysis, degradation should be accomplished via destruction of Adda group by oxidation, with the formation of dihydroxy substituendums as end products. This study has provided a fundamental demonstration of C102 serving as oxidizing disinfectant to eliminate microcystins from raw water source.