ABSTRACT
Long-term overgrazing may lead to the degradation of grasslands which are often characterized by an increase in nonpreferred species, especially toxic plants. However, the impact of these toxic nonpreferred species on the restoration processes of degraded grasslands is not well understood, particularly their interactions with soil properties and other plant functional groups. To address this knowledge gap, we conducted an in situ grazing exclusion experiment in a temperate degraded grassland of Inner Mongolia, China. The objective of this study was to investigate how toxic nonpreferred plants influence the recovery of plant diversity and productivity in degraded grasslands and whether these effects can be explained by changes in soil properties. Our findings revealed that Stellera chamaejasme, a toxic nonpreferred species widely distributed in North China, directly altered plant community composition and improved species diversity in degraded grasslands dominated by Asteraceae plants. The presence of S. chamaejasme could inhibit Asteraceae abundance and increase soil copper content in this study area, because Asteraceae plants have a high copper accumulation capacity. Within the communities with S. chamaejasme, the alleviation of soil copper limitation to plants may subsequently enhance the abundance and aboveground productivity of Poaceae and Forbs. Our study demonstrated that the strong direct and indirect interactions of toxic nonpreferred species with other ecosystem components promoted competitive release in terms of biomass accumulation and species diversity. The change of soil limiting microelements content caused by toxic species exerts an important mediation function during the recovery process of degraded grasslands. Thus, these toxic nonpreferred species can act primarily as accelerators for the restoration of community structure and ecosystem function in degraded grasslands.
Subject(s)
Biodiversity , Grassland , China , Soil/chemistry , Poaceae , Asteraceae/physiologyABSTRACT
Oral diseases concern almost every individual and are a serious health risk to the population. The restorative treatment of tooth and jaw defects is an important means to achieve oral function and support the appearance of the contour. Based on the principle of "learning from the nature", Deng Xuliang's group of Peking University School and Hospital of Stomatology has proposed a new concept of "microstructural biomimetic design and tissue adaptation of tooth/jaw materials" to address the worldwide problems of difficulty in treating dentine hypersensitivity, poor prognosis of restoration of tooth defects, and vertical bone augmentation of alveolar bone after tooth loss. The group has broken through the bottleneck of multi-stage biomimetic technology from the design of microscopic features to the enhancement of macroscopic effects, and invented key technologies such as crystalline/amorphous multi-level assembly, ion-transportation blocking, and multi-physical properties of the micro-environment reconstruction, etc. The group also pioneered the cationic-hydrogel desensitizer, digital stump and core integrated restorations, and developed new crown and bridge restorative materials, gradient functionalisation guided tissue regeneration membrane, and electrically responsive alveolar bone augmentation restorative membranes, etc. These products have established new clinical strategies for tooth/jaw defect repair and achieved innovative results. In conclusion, the research results of our group have strongly supported the theoretical improvement of stomatology, developed the technical system of oral hard tissue restoration, innovated the clinical treatment strategy, and led the progress of the stomatology industry.
Subject(s)
Bionics , Dental Restoration, Permanent , Mouth Diseases , HumansABSTRACT
Vertical greenery system (VGS) is a sustainable solution to promote building energy saving and emission reduction, mitigate the urban heat island effect, as well as a crucial component of urban ecological construction. We summarized four main mechanisms of the thermal effects of VGSs, including shading effect, evapotranspiration effect, thermal insulation effect, and wind control effect. We elucidated the effects of VGSs on building cooling and energy saving, and analyzed the cooling effects of VGSs on plant canopy and outdoor ambient air, as well as their influence on mitigating the urban heat island effect. Based on available research on the thermal effects of VGSs, we identified key directions for future research, aiming to expedite the development of green cities and achieve carbon neutrality.
Subject(s)
Cold Temperature , Hot Temperature , Cities , Wind , PlantsABSTRACT
The heterogeneity of extracellular matrix (ECM) topology, stiffness, and architecture is a key factor modulating cellular behavior and osteogenesis. However, the effects of heterogeneous ECM electric potential at the micro- and nanoscale on osteogenesis remain to be elucidated. Here, the heterogeneous distribution of surface potential is established by incorporating ferroelectric BaTiO3 nanofibers (BTNF) into poly(vinylidene fluoridetrifluoroethylene) (P(VDF-TrFE)) matrix based on phase-field and first-principles simulation. By optimizing the aspect ratios of BTNF fillers, the anisotropic distribution of surface potential on BTNF/P(VDF-TrFE) nanocomposite membranes can be achieved by strong spontaneous electric polarization of BTNF fillers. These results indicate that heterogeneous surface potential distribution leads to a meshwork pattern of fibronectin (FN) aggregation, which increased FN-III7-10 (FN fragment) focal flexibility and anchor points as predicted by molecular dynamics simulation. Furthermore, integrin clustering, focal adhesion formation, cell spreading, and adhesion are enhanced sequentially. Increased traction of actin fibers amplifies mechanotransduction by promoting nuclear translocation of YAP/Runx2, which enhances osteogenesis in vitro and bone regeneration in vivo. The work thus provides fundamental insights into the biological effects of surface potential heterogeneity at the micro- and nanoscale on osteogenesis, and also develops a new strategy to optimize the performance of electroactive biomaterials for tissue regenerative therapies.
Subject(s)
Mechanotransduction, Cellular , Osteogenesis , Cell Differentiation , Bone Regeneration , Fibronectins/pharmacologyABSTRACT
Correction for 'Restoring the electrical microenvironment using ferroelectric nanocomposite membranes to enhance alveolar ridge regeneration in a mini-pig preclinical model' by Yiping Li et al., J. Mater. Chem. B, 2023, 11, 985-997, https://doi.org/10.1039/D2TB02054H.
ABSTRACT
Many studies have focused mainly on the thermal and energy performance of VGSs on a building scale; however, little is known about the cooling effect of VGSs in street canyons and its response to different aspect ratios (building height/street width, H/W). Therefore, a scaled outdoor experiment was conducted to investigate the spatiotemporal variation of the urban wind and thermal environment caused by west-facing vertical greenery systems (VGSs) in street canyons with H/W = 1 and 2 in the subtropical city of Guangzhou, China. On a typical hot day (26th Oct. 2021), VGSs reduced wind speed by 38.0 % and 21.0 % in street canyons with H/W = 1 and 2, respectively. Compared with the reference cases, the temperature regimes of VGS cases (H/W = 1 and 2), including west-facing wall temperature (Tw-west), east-facing wall temperature (Tw-east), ground temperature (Tg), and air temperature (Ta), were all significantly decreased. Because of the cooling effect of VGSs, the maximum reduction of Tw-west in the upper level of street canyons with H/W = 1 and 2 was 20.3 and 16.8 °C, respectively. The maximum reduction of Tg in the center of the VGS case with H/W = 2 was 4.6 °C, which was more pronounced than in the VGS case with H/W = 1 (1.8 °C). The maximum reduction of Ta at the pedestrian level along the central axis of street canyons with H/W = 1 and 2 was 0.8 and 1.6 °C, respectively, which was more pronounced than at the upper level. The reduction of Tw-west in the upper level and of Tg and Ta in the VGS case with H/W = 2 was greater than that in the VGS case with H/W = 1 due to lower wind speed and albedo.
Subject(s)
Air Pollutants , Pedestrians , Humans , Models, Theoretical , Wind , Cities , Temperature , Air Pollutants/analysisABSTRACT
The maintenance and incremental growth of the alveolar bone at the tooth extraction site, to achieve the required height and width for implant restoration, remains a major clinical challenge. Here, the concept of restoring the electrical microenvironment to improve the effects of alveolar ridge preservation (ARP) was investigated in a mini-pig preclinical model. The endogeneous electrical microenvironment of the dental alveolar socket was recapitulated by fabricating a biomimetic ferroelectric BaTiO3/poly(vinylidene fluoridetrifluoroethylene) (BTO/P(VDF-TrFE)) non-resorbable nanocomposite membrane polarized by corona poling. The polarized nanocomposite membrane exhibited excellent electrical stability. After implantation with bone grafts and covering with the charged membrane in tooth extraction sites for three months, both the vertical and horizontal dimension resorption of the alveolar ridge were significantly prevented, as assessed by cone beam computed tomography (CBCT) analyses. Micro-CT analysis showed that the charged membrane induced significant enhancement of newly regenerated bone at the tooth extraction sites. Histological analysis further confirmed that the restoration of the electrical microenvironment significantly promoted buccal alveolar bone regeneration and maturation. In addition, the charged membranes can maintain their structural integrity during the entire implantation period and exhibit positive long-term systemic safety, as assessed by preclinical sub-chronic systemic toxicity. These findings thus provide an innovative strategy for restoring the electrical microenvironment to enhance ARP following dentition defect and edentulism, which could further advance prosthodontics implant technology.
Subject(s)
Nanocomposites , Tooth Socket , Animals , Swine , Swine, Miniature , Tooth Socket/pathology , Alveolar Process/pathology , Bone RegenerationABSTRACT
Pyrite is one kind of cost-effective electron donors for nitrate denitrification. In this study, a pyrite-driven autotrophic denitrification biofilter was applied for simultaneous removal of NH4+ and NO3- over the 150-day. The influent NH4+/NO3- ratio (0.3-1.7) had less effect on system performance, while for the hydraulic retention times (HRTs, 24-3 h), the removal percentage of both > 90% and removal loading rates of 52.8 and 59.4 mg N/(L·d) for NH4+ and NO3- removal were obtained at the HRT of 6 h. The 16S rRNA genes analysis showed that Ferritrophicum, Thiobacillus, Candidatus_Brocadia, and unidentified_Nitrospiraceae were predominant. Analyses of nitrogen and sulfur metabolism showed that ammonia was removed by complete nitrification, nitrate was reduced to N2, and sulfide was oxidized to sulfate. Dynamics of pollutants within the reactor and microbial activity showed nitrification/Anammox and pyrite-driven autotrophic denitrification were responsible for the synergistic removal of NH4+/NO3- in this system.
Subject(s)
Denitrification , Nitrates , Ammonia , Autotrophic Processes , Bioreactors , Iron , Nitrogen , Oxidation-Reduction , RNA, Ribosomal, 16S/genetics , SulfidesABSTRACT
The gene activity underlying cell differentiation is regulated by a diverse set of transcription factors (TFs), histone modifications, chromatin structures and more. Although definitive hematopoietic stem cells (HSCs) are known to emerge via endothelial-to-hematopoietic transition (EHT), how the multi-layered epigenome is sequentially unfolded in a small portion of endothelial cells (ECs) transitioning into the hematopoietic fate remains elusive. With optimized low-input itChIP-seq and Hi-C assays, we performed multi-omics dissection of the HSC ontogeny trajectory across early arterial ECs (eAECs), hemogenic endothelial cells (HECs), pre-HSCs and long-term HSCs (LT-HSCs) in mouse embryos. Interestingly, HSC regulatory regions are already pre-configurated with active histone modifications as early as eAECs, preceding chromatin looping dynamics within topologically associating domains. Chromatin looping structures between enhancers and promoters only become gradually strengthened over time. Notably, RUNX1, a master TF for hematopoiesis, enriched at half of these loops is observed early from eAECs through pre-HSCs but its enrichment further increases in HSCs. RUNX1 and co-TFs together constitute a central, progressively intensified enhancer-promoter interactions. Thus, our study provides a framework to decipher how temporal epigenomic configurations fulfill cell lineage specification during development.
Subject(s)
Chromatin/chemistry , Embryo, Mammalian/cytology , Hematopoietic Stem Cells/cytology , Histone Code , Animals , Cluster Analysis , Core Binding Factor Alpha 2 Subunit/metabolism , Enhancer Elements, Genetic/genetics , Genome , Mice, Inbred C57BL , Molecular Sequence Annotation , Promoter Regions, Genetic/genetics , Transcription Factors/metabolismABSTRACT
Ferroelectric biomaterials have been widely investigated and demonstrated to enhance osteogenesis by simulating the inherent electrical properties of bone tissues. Nevertheless, the underlying biological processes are still not wellunderstood. Hence, this study investigated the underlying biological processes by which bone piezoelectricity-mimicking barium titanate/poly(vinylidene fluoride-trifluoroethylene) nanocomposite membranes (BTO nanocomposite membranes) promote osteogenesis of Bone Marrow Mesenchymal Stem Cells (BMSCs). Ourresults revealed that the piezoelectric coefficient (d33) of nanocomposite membranes aftercontrolled corona poling was similar to that of native bone, and exhibited highly-stable piezoelectrical properties and concentrated surface electrical potential. These nanocomposite membranes significantly enhanced the adhesion and spreading of BMSCs, which was manifested as increased number and area of mature focal adhesions. Furthermore, the nanocomposite membranes significantly promoted the expression of integrin receptors genes (α1, α5 andß3), which in turn enhanced osteogenesis of BMSCs, as manifested by upregulated Alkaline Phosphatase (ALP) and Bone Morphogenetic Protein 2 (BMP2) expression levels. Further investigations found that the Focal Adhesion Kinase (FAK)-Extracellular Signal-Regulated Kinase1/2 (ERK 1/2) signaling axis may be involved in the biological process of polarized nanocomposite membrane-induced osteogenesis. This study thus provides useful insights for betterunderstanding of the biological processes by which piezoelectric or ferroelectric biomaterials promote osteogenesis.
Subject(s)
Mesenchymal Stem Cells , Nanocomposites , Actin Cytoskeleton , Bone Marrow Cells , Bone and Bones , Cell Adhesion , Cell Differentiation , Cells, Cultured , OsteogenesisABSTRACT
Immune dysfunction is implicated in Alzheimer's disease (AD), whereas systemic immune modulation may be neuroprotective. Our previous results have indicated immune challenge with Bacillus Calmette-Guerin attenuates AD pathology in animal models by boosting the systemic immune system. Similarly, independent studies have shown that boosting systemic immune system, by blocking PD-1 checkpoint pathway, modifies AD. Here we hypothesized that influenza vaccine would potentiate function of moderate dose anti-PD-1 and therefore combining them might allow reducing the dose of PD-1 antibody needed to modify the disease. We found that moderate-dose PD-1 in combination with influenza vaccine effectively attenuated cognitive deficit and prevented amyloid-ß pathology build-up in APP/PS1 mice in a mechanism dependent on recruitment of peripheral monocyte-derived macrophages into the brain. Eliminating peripheral macrophages abrogated the beneficial effect. Moreover, by comparing CD11b+ compartments in the mouse parenchyma, we observed an elevated subset of Ly6C+ microglia-like cells, which are reportedly derived from peripheral monocytes. In addition, myeloid-derived suppressor cells are strongly elevated in the transgenic model used and normalized by combination treatment, indicating restoration of brain immune homeostasis. Overall, our results suggest that revitalizing brain immunity by combining IV with moderate-dose PD-1 inhibition may represent a therapeutic immunotherapy for AD.
Subject(s)
Alzheimer Disease , Influenza Vaccines , Amyloid beta-Peptides/metabolism , Amyloid beta-Protein Precursor/genetics , Amyloid beta-Protein Precursor/metabolism , Animals , Brain/metabolism , Cognition , Disease Models, Animal , Mice , Mice, Inbred C57BL , Mice, Transgenic , Presenilin-1ABSTRACT
Hematopoietic stem cells (HSCs) in adults are believed to be born from hemogenic endothelial cells (HECs) in mid-gestational embryos. Due to the rare and transient nature, the HSC-competent HECs have never been stringently identified and accurately captured, let alone their genuine vascular precursors. Here, we first used high-precision single-cell transcriptomics to unbiasedly examine the relevant EC populations at continuous developmental stages with intervals of 0.5 days from embryonic day (E) 9.5 to E11.0. As a consequence, we transcriptomically identified two molecularly different arterial EC populations and putative HSC-primed HECs, whose number peaked at E10.0 and sharply decreased thereafter, in the dorsal aorta of the aorta-gonad-mesonephros (AGM) region. Combining computational prediction and in vivo functional validation, we precisely captured HSC-competent HECs by the newly constructed Neurl3-EGFP reporter mouse model, and realized the enrichment further by a combination of surface markers (Procr+Kit+CD44+, PK44). Surprisingly, the endothelial-hematopoietic dual potential was rarely but reliably witnessed in the cultures of single HECs. Noteworthy, primitive vascular ECs from E8.0 experienced two-step fate choices to become HSC-primed HECs, namely an initial arterial fate choice followed by a hemogenic fate conversion. This finding resolves several previously observed contradictions. Taken together, comprehensive understanding of endothelial evolutions and molecular programs underlying HSC-primed HEC specification in vivo will facilitate future investigations directing HSC production in vitro.
Subject(s)
Aorta/embryology , Hemangioblasts/cytology , Hematopoiesis , Transcriptome , Animals , Cells, Cultured , Embryo, Mammalian , Mice , Mice, Inbred C57BL , Primary Cell Culture , Single-Cell AnalysisABSTRACT
The functional heterogeneity of hematopoietic stem cells (HSCs) has been comprehensively investigated by single-cell transplantation assay. However, the heterogeneity regarding their physiological contribution remains an open question, especially for those with life-long hematopoietic fate of rigorous self-renewing and balanced differentiation capacities. In this study, we revealed that Procr expression was detected principally in phenotypical vascular endothelium co-expressing Dll4 and CD44 in the mid-gestation mouse embryos, and could enrich all the HSCs of the embryonic day 11.5 (E11.5) aorta-gonad-mesonephros (AGM) region. We then used a temporally restricted genetic tracing strategy to irreversibly label the Procr-expressing cells at E9.5. Interestingly, most labeled mature HSCs in multiple sites (such as AGM) around E11.5 were functionally categorized as lymphomyeloid-balanced HSCs assessed by direct transplantation. Furthermore, the labeled cells contributed to an average of 7.8% of immunophenotypically defined HSCs in E14.5 fetal liver (FL) and 6.9% of leukocytes in peripheral blood (PB) during one-year follow-up. Surprisingly, in aged mice of 24 months, the embryonically tagged cells displayed constant contribution to leukocytes with no bias to myeloid or lymphoid lineages. Altogether, we demonstrated, for the first time, the existence of a subtype of physiologically long-lived balanced HSCs as hypothesized, whose precise embryonic origin and molecular identity await further characterization.
Subject(s)
Endothelial Protein C Receptor/metabolism , Hematopoietic Stem Cells/metabolism , Animals , Aorta/cytology , Aorta/metabolism , Embryo, Mammalian , Endothelial Protein C Receptor/genetics , Female , Hematopoiesis/genetics , Hematopoiesis/physiology , Hematopoietic Stem Cells/cytology , Male , Mesonephros/cytology , Mesonephros/metabolism , Mice , Platelet Endothelial Cell Adhesion Molecule-1/genetics , Platelet Endothelial Cell Adhesion Molecule-1/metabolismABSTRACT
BACKGROUND: Prenatal infection is a substantial risk factor for neurodevelopmental disorders such as autism in offspring. We have previously reported that influenza vaccination (VAC) during early pregnancy contributes to neurogenesis and behavioral function in offspring. RESULTS: Here, we probe the efficacy of VAC pretreatment on autism-like behaviors in a lipopolysaccharide (LPS)-induced maternal immune activation (MIA) mouse model. We show that VAC improves abnormal fetal brain cytoarchitecture and lamination, an effect associated with promotion of intermediate progenitor cell differentiation in MIA fetal brain. These beneficial effects are sufficient to prevent social deficits in adult MIA offspring. Furthermore, whole-genome analysis suggests a strong interaction between Ikzf1 (IKAROS family zinc-finger 1) and neuronal differentiation. Intriguingly, VAC rescues excessive microglial Ikzf1 expression and attenuates microglial inflammatory responses in the MIA fetal brain. CONCLUSIONS: Our study implies that a preprocessed influenza vaccination prevents maternal bacterial infection from causing neocortical lamination impairments and autism-related behaviors in offspring.
Subject(s)
Autistic Disorder/complications , Influenza Vaccines/therapeutic use , Malformations of Cortical Development/prevention & control , Prenatal Exposure Delayed Effects/physiopathology , Social Behavior Disorders/prevention & control , Animals , Animals, Newborn , CX3C Chemokine Receptor 1/genetics , CX3C Chemokine Receptor 1/metabolism , Disease Models, Animal , Exploratory Behavior/drug effects , Exploratory Behavior/physiology , Female , Gene Expression Regulation/drug effects , Gene Expression Regulation/genetics , Lipopolysaccharides/toxicity , Male , Malformations of Cortical Development/etiology , Maze Learning/drug effects , Mice , Mice, Inbred C57BL , Mice, Transgenic , Pregnancy , Prenatal Exposure Delayed Effects/chemically induced , Social Behavior Disorders/etiology , Swimming/physiology , Swimming/psychologyABSTRACT
Haematopoietic stem cells (HSCs) are derived early from embryonic precursors, such as haemogenic endothelial cells and pre-haematopoietic stem cells (pre-HSCs), the molecular identity of which still remains elusive. Here we use potent surface markers to capture the nascent pre-HSCs at high purity, as rigorously validated by single-cell-initiated serial transplantation. Then we apply single-cell RNA sequencing to analyse endothelial cells, CD45(-) and CD45(+) pre-HSCs in the aorta-gonad-mesonephros region, and HSCs in fetal liver. Pre-HSCs show unique features in transcriptional machinery, arterial signature, metabolism state, signalling pathway, and transcription factor network. Functionally, activation of mechanistic targets of rapamycin (mTOR) is shown to be indispensable for the emergence of HSCs but not haematopoietic progenitors. Transcriptome data-based functional analysis reveals remarkable heterogeneity in cell-cycle status of pre-HSCs. Finally, the core molecular signature of pre-HSCs is identified. Collectively, our work paves the way for dissection of complex molecular mechanisms regulating stepwise generation of HSCs in vivo, informing future efforts to engineer HSCs for clinical applications.
Subject(s)
Cell Differentiation , Cell Tracking/methods , Hematopoietic Stem Cells/cytology , Single-Cell Analysis/methods , Animals , Aorta/cytology , Biomarkers/analysis , Cell Cycle/genetics , Cell Differentiation/genetics , Cell Lineage , Endothelial Cells/cytology , Endothelial Cells/metabolism , Endothelial Protein C Receptor , Female , Fetus/cytology , Gonads/cytology , Hematopoietic Stem Cells/metabolism , Leukocyte Common Antigens/analysis , Leukocyte Common Antigens/metabolism , Liver/cytology , Male , Mechanistic Target of Rapamycin Complex 2 , Mesonephros/cytology , Mice , Multiprotein Complexes/metabolism , Receptors, Cell Surface/metabolism , Reproducibility of Results , Sequence Analysis, RNA , Signal Transduction , TOR Serine-Threonine Kinases/metabolism , Transcription Factors/metabolism , Transcription, Genetic/genetics , TranscriptomeABSTRACT
Cannabinoid receptor interacting protein 1 (CNRIP1), which has been originally identified as the binding partner of cannabinoid receptor 1 (CNR1), is evolutionarily conserved throughout vertebrates, but its physiological function has been unknown. Here, we identify a developmental role of CNRIP1 using Xenopus laevis embryos. During early embryogenesis, expression of Xenopus laevis cnrip1 is highly restricted to the animal region of gastrulae where neural and eye induction occur, and afterward it is seen in neural and other tissues with a temporally and spatially regulated pattern. Morpholino-mediated knockdown experiments indicate that cnrip1 has an essential role in early eye and neural development by regulating the onset of expression of key transcription factor genes, sox2, otx2, pax6 and rax. Also, over-expression experiments suggest that cnrip1 has a potential to expand sox2, otx2, pax6 and rax expression. These results suggest an instructive role of Xenopus laevis cnrip1 in early eye and neural development. Furthermore, Xenopus laevis cnr1 knockdown leads to eye defects, which are partly similar to, but milder than, those caused by cnrip1 knockdown, suggesting a possible functional similarity between CNRIP1 and CNR1. This study is the first characterization of an in vivo role of CNRIP1 in the context of whole organisms.
Subject(s)
Carrier Proteins/metabolism , Eye/embryology , Gastrula/metabolism , Neurogenesis , Xenopus Proteins/metabolism , Xenopus laevis/embryology , Xenopus laevis/metabolism , Animals , Carrier Proteins/genetics , Eye/metabolism , Transcription Factors/metabolism , Xenopus Proteins/geneticsABSTRACT
Some reports show that lanthanum, a rare earth element, induces apoptosis in certain cancer cells. In the present paper, we report the first observation of anoikis induced by lanthanum citrate (LaCit) in Hela cells at a concentration of 0.001-0.1 mmol/L after 48h-treatment. Before cell treatment, Hela cells were subjected to anoikis-resistant selection to remove anoikis-sensitive cells and ensure specificity of LaCit-induced anoikis. Anoikis was determined by Annexin/PI, AO/EB staining, cleavage of PARP and soft-agar colony forming assay. Further, findings of decreased mitochondrial membrane potential, the cleavage of caspase-9 and a dose-dependent increase expression of Bax were detected, suggesting that the intrinsic caspase pathway was involved in the anoikis induced by LaCit. In addition, activation of caspase-8 occurred later than that of caspase-9. LaCit also caused reorganization of actin cytoskeleton, and was accompanied by an increase in co-localization of F-actin with mitochondria, implying that both actin cytoskeleton and mitochondria may play important roles in LaCit -induced anoikis.