Identification of novel targets and mechanisms of wogonin on lung cancer,bladder cancer,and colon cancer

Wogonin(WOG)has been demonstrated to have anti-cancer activity,but the mechanisms remain unclear.In this study,new targets of WOG were predicted for lung cancer,bladder cancer,and colon cancer by using bioinformatics methods.wOG might primarily suppress cancers via regulating arachidonic acid,Ras,MAPK,linoleic acid,PI3K Akt,and folate biosynthesis pathways.3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)assay showed that WOG inhibited the proliferation of A549 cells.Real-time quantitative reverse transcription PCR(RT-qPCR)results indicated that anti-lung cancer effect of WOG was achieved by regulating the expression of 18 target genes,including AKRIBI0,AKRIC3,BDNF,CAVI,CXCL2,CYP2B6,CYP4F3,DAO,EGF,ENO3,IL6,PLA2GlB,PLA2G2F,PLA2G4A,PTGES,SLCOIBI,SLCOIB3,and TFAP2A.The Kaplan-Meier survival curves further confirmed that DAO,PLA2G1B,SLCO1B3 and TFAP2A were essential targets via which WOG affected lung cancer survival.Moreover,BDNF,FGF2,and PTGS1 were predicted to be the targets via which WOG alleviated cancer proliferation and invasion in bladder cancer.As for colon cancer,WOG might induce autophagy and inhibit proliferation by down-regulating NTF4 and TH.The study will provide clue for using wOG as a promising antineoplastic agent in basic and translational research,and bring light to the application of herbs containing WOG as food supplements.

Graded and pan-neural disease phenotypes of Rett Syndrome linked with dosage of functional MeCP2

Rett syndrome(RTT)is a progressive neurodevelop-mental disorder,mainly caused by mutations in MeCP2 and currently with no cure.We report here that neurons from R106W MeCP2 RTT human iPSCs as well as human embryonic stem cells after MeCP2 knockdown exhibit consistent and long-lasting impairment in maturation as indicated by impaired action potentials and passive membrane properties as well as reduced soma size and spine density.Moreover,RTT-inherent defects in neuronal maturation could be pan-neuronal and occurred in neurons with both dorsal and ventral forebrain features.Knockdown of MeCP2 led to more severe neuronal deficits as compared to RTT iPSC-derived neurons,which appeared to retain partial function.Strikingly,consistent deficits in nuclear size,dendritic complexity and circuitry-dependent spontaneous postsynaptic currents could only be observed in MeCP2 knockdown neurons but not RTT iPSC-derived neurons.Both neuron-intrinsic and circuitry-dependent deficits of MeCP2-deficient neurons could be fully or partially rescued by re-expression of wild type or T158M MeCP2,strengthening the dosage dependency of MeCP2 on disease phenotypes and also the partial function of the mutant.Our findings thus reveal stable neuronal maturation deficits and unexpectedly,graded sensitivities of neuron-inherent and neural transmission phenotypes towards the extent of MeCP2 deficiency,which is informative for future therapeutic development.

Ethyl Acetate Fraction in Hedyotis Diffusa Willd Inhibits T Cell Proliferation to Improve the Pathogenesis of Systemic Lupus Erythematosus

Background:Abnormal proliferation of T cells plays an essential role in the pathogenesis of Systemic lupus erythematosus(SLE).The pharmaceutical effect of Hedyotis Diffusa Willd(HDW)on SLE has been investigated previously.Nevertheless,the biomedical mechanism is still left unclear.Objective:This study has been arranged to evaluate the therapeutic effect of the ethyl acetate fraction of HDW(EAHDW)on lupus mice and explore the potential therapeutic mechanism.Methods:EAHDW was prepared with 80%ethanol reflex extraction followed by successive extraction,and ana-lyzed with HPLC and UPLC-Q/TOP-MS.The potential targets and STAT3 affinity regulators were predicted with network pharmacology.The pharmaceutic effect of EAHDW was studied with MRL/lpr mice.Cytokines and au-toantibodies were quantified with ELISA assays.The pathological damage of glomerulus and STAT3 expression in the kidney was detected with histochemical and immunohistochemical techniques.The cell cycle properties in cell proliferation were identified with the flow cytometry.The western blot and dual-Luciferase reporter assay were applied to evaluate translational and transcriptional activity of STAT3,respectively.Results:In this study,the extraction ratio of EAHDW was 2.7±1%,in which 19 ingredients were identified.Network pharmacological analysis showed that the target genes of EAHDW were highly focused on influencing the abnormal T cell proliferation in SLE.EAHDW showed the beneficial effects on pathological changes and STAT3 expression in the glomerulus of lupus mice,and the levels of cytokines and autoantibodies in serum.In cytological study,EAHDW treatment attenuated the transcription and phosphorylation of STAT3,which inhibited T cell proliferation by prolonged S-phase of the cell cycle.A total of 5 compounds in EAHDW exhibited high docking affinity to the DNA-binding site of STAT3.Conclusion:EAHDW could reduce the inflammatory response and inhibit the proliferation of T cells by interfering with the STAT3 signaling pathway,thereby playing a therapeutic effect on SLE.

Temporal and spatial cellular and molecular pathological alterations with single-cell resolution in the adult spinal cord after injury

Spinal cord injury(SCI)involves diverse injury responses in different cell types in a temporally and spatially specific manner.Here,using single-cell transcriptomic analyses combined with classic anatomical,behavioral,electrophysiological analyses,we report,with single-cell resolution,temporal molecular and cellular changes in crush-injured adult mouse spinal cord.Data revealed pathological changes of 12 different major cell types,three of which infiltrated into the spinal cord at distinct times post-injury.We discovered novel microglia and astrocyte subtypes in the uninjured spinal cord,and their dynamic conversions into additional stage-specific subtypes/states.Most dynamic changes occur at 3-days post-injury and by day-14 the second wave of microglial activation emerged,accompanied with changes in various cell types including neurons,indicative of the second round of attacks.By day-38,major cell types are still substantially deviated from uninjured states,demonstrating prolonged alterations.This study provides a comprehensive mapping of cellular/molecular pathological changes along the temporal axis after SCI,which may facilitate the development of novel therapeutic strategies,including those targeting microglia.

Unbiased transcriptomic analyses reveal distinet effects of immune deficiency in CNS function with and without injury

The mammalian central nervous system (CNS) is considered an immune privileged system as it is separated from the periphery by the blood brain barrier (BBB). Yet, immune functions have been postulated to heavily influence the functional state of the CNS, especially after injury or during neurodegeneration. There is controversy regarding whether adaptive immune responses are beneficial or detrimental to CNS injury repair. In this study, we utilized immunocompromised SCID mice and subjected them to spinal cord injury (SCI). We analyzed motor function, electrophysiology, histochemistry, and performed unbiased RNA-sequencing. SCID mice displayed improved CNS functional recovery compared to WT mice after SCI. Weighted gene-coexpression network analysis (WGCNA) of spinal cord transcriptomes revealed that SCID mice had reduced expression of immune function-related genes and heightened expression of neural transmission-related genes after SCI, which was confirmed by immunohistochemical analysis and was consistent with better functional recovery. Transcriptomic analyses also indicated heightened expression of neurotransmission-related genes before injury in SCID mice, suggesting that a steady state of immune-deficiency potentially led to CNS hyper-connectivity. Consequently, SCID mice without injury demonstrated worse performance in Morris water maze test. Taken together, not only reduced inflammation after injury but also dampened steady-state immune function without injury heightened the neurotransmission program, resulting in better or worse behavioral outcomes respectively. This study revealed the intricate relationship between immune and nervous systems, raising the possibility for therapeutic manipulation of neural function via immune modulation.