Alzheimer's disease(AD),a progressive dementia,is one of the most common neurodegenerative dis-eases.Clinical trial results of amyloid-β(Aβ)and tau regulators based on the pretext of straightforward amyloid and tau immunotherapy were disappointing.There are currently no effective strategies for slowing the progression of AD.Herein,we spotlight the dysregulation of lipid metabolism,particularly the elevation of ceramides(Cers),as a critical yet underexplored facet of AD pathogenesis.Our study delineates the role of Cers in promoting microglial pyroptosis,a form of programmed cell death distinct from apoptosis and necroptosis,characterized by cellular swelling,and membrane rupture mediated by the NLRP3 inflammasome pathway.Utilizing both in vivo experiments with amyloid precursor protein(APP)/presenilin 1(PS1)transgenic mice and in vitro assays with BV-2 microglial cells,we investigate the activation of microglial pyroptosis by Cers and its inhibition by icariin(ICA),a flavonoid with known antioxidant and anti-inflammatory properties.Our findings reveal a significant increase in Cers levels and pyroptosis markers(NOD-like receptor family,pyrin domain containing 3(NLRP3),apoptosis-associated speck-like protein containing a caspase recruitment domain,caspase-1,gasdermin D(GSDMD),and interleukin-18(IL-18))in the brains of AD model mice,indicating a direct involvement of Cers in AD pathology through the induction of microglial pyroptosis.Conversely,ICA treatment effec-tively reduces these pyroptotic markers and Cer levels,thereby attenuating microglial pyroptosis and suggesting a novel therapeutic mechanism of action against AD.This study not only advances our un-derstanding of the pathogenic role of Cers in AD but also introduces ICA as a promising candidate for AD therapy,capable of mitigating neuroinflammation and pyroptosis through the cyclooxygenase-2(COX-2)-NLRP3 inflammasome-gasdermin D(GSDMD)axis.Our results pave the way for further exploration of Cer metabolism disorders in neurodegenerative diseases and highlight the therapeutic potential of tar-geting microglial pyroptosis in AD.

OBJECTIVES: To explore the correlation of ELFN1 expression level with prognosis of colorectal cancer and its regulatory role in colorectal cancer cell proliferation and metastasis. METHODS: We analyzed the expression levels of ELFN1 across 33 cancer types using publicly available databases and identified differential genes related to ELFN1 in colorectal cancer. Gene function annotation and enrichment analysis were used to identify the involved signaling pathways. Logistic analysis, Kaplan-Meier analysis and Cox regression analysis were performed to evaluate the correlation between ELFN1 expression and clinicopathological parameters and survival of colorectal cancer patients. qPCR and Western blotting were used to validate the expression levels of ELFN1 in different colorectal cancer cell lines and tissues, and Transwell and EDU experiments were carried out to assess the effect of ELFN1 knockdown on biological behaviors of SW480 cells. RESULTS: ELFN1 was highly expressed in 14 cancers, and its expression was significantly higher in colon cancer tissues than in adjacent tissues. A high expression of ELFN1 mRNA was associated with a poorer overall survival of colorectal cancer patients. Cox regression analysis indicated that ELFN1 expression was an independent prognostic factor for overall survival of the patients. ELFN1 was significantly enriched in tumor metastasis and proliferation and participated in several tumor signaling pathways. The colon cancer cell lines showed significantly higher expression levels of ELFN1 than normal cells, ELFN1 knockdown obviously inhibited proliferation and migration of SW480 cells in vitro. CONCLUSIONS ELFN1 is overexpressed in colorectal cancer and is associated with poor clinical prognosis of the patients. A high ELFN1 expression is associated with malignant phenotypes of colorectal cancer and promotes cancer cell proliferation and metastasis, suggesting its potential as a prognostic biomarker for colorectal cancer.
Humans ; Colorectal Neoplasms/diagnosis* ; Cell Proliferation ; Prognosis ; Cell Line, Tumor ; Biomarkers, Tumor/metabolism* ; Neoplasm Metastasis ; Gene Expression Regulation, Neoplastic ; Nerve Tissue Proteins/metabolism* ; Female ; Male

Alzheimer's disease (AD), a progressive dementia, is one of the most common neurodegenerative diseases. Clinical trial results of amyloid-β (Aβ) and tau regulators based on the pretext of straightforward amyloid and tau immunotherapy were disappointing. There are currently no effective strategies for slowing the progression of AD. Herein, we spotlight the dysregulation of lipid metabolism, particularly the elevation of ceramides (Cers), as a critical yet underexplored facet of AD pathogenesis. Our study delineates the role of Cers in promoting microglial pyroptosis, a form of programmed cell death distinct from apoptosis and necroptosis, characterized by cellular swelling, and membrane rupture mediated by the NLRP3 inflammasome pathway. Utilizing both in vivo experiments with amyloid precursor protein (APP)/presenilin 1 (PS1) transgenic mice and in vitro assays with BV-2 microglial cells, we investigate the activation of microglial pyroptosis by Cers and its inhibition by icariin (ICA), a flavonoid with known antioxidant and anti-inflammatory properties. Our findings reveal a significant increase in Cers levels and pyroptosis markers (NOD-like receptor family, pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein containing a caspase recruitment domain, caspase-1, gasdermin D (gasdermin D (GSDMD)), and interleukin-18 (IL-18)) in the brains of AD model mice, indicating a direct involvement of Cers in AD pathology through the induction of microglial pyroptosis. Conversely, ICA treatment effectively reduces these pyroptotic markers and Cer levels, thereby attenuating microglial pyroptosis and suggesting a novel therapeutic mechanism of action against AD. This study not only advances our understanding of the pathogenic role of Cers in AD but also introduces ICA as a promising candidate for AD therapy, capable of mitigating neuroinflammation and pyroptosis through the cyclooxygenase-2 (COX-2)-NLRP3 inflammasome-gasdermin D (GSDMD) axis. Our results pave the way for further exploration of Cer metabolism disorders in neurodegenerative diseases and highlight the therapeutic potential of targeting microglial pyroptosis in AD.

Objective To explore the effects of family functioning,peer victimization and school connectedness on nonsuicidal self-injury in adolescents,as well as the mediating effect of psychache in these relationships.Methods A total of 525 junior high school students in Foshan City were surveyed using the brief Family Function Scale,Peer Victimization Scale,School Connectedness Scale,Psychache Scale,and Nonsuicidal Self-injury Scale.Descriptive statistics and correlation analyses were performed using SPSS 23.0,and mediation analysis was conducted using Mplus 7.4.Results ① Family functioning could directly predict adolescent non-suicidal self-injury(β=-0.20,P＜0.01),but did not exert an indirect effect through psychache(P=0.123).② Peer victimization could not directly predict adolescent non-suicidal self-injury(β=0.10,P=0.133),but had an indirect effect through psychache,with an indirect effect value of 0.142(P＜0.001).③ School connectedness could not directly predict adolescent non-suicidal self-injury(β=0.13,P=0.053),but had an indirect effect through psychache,with an indirect effect value of-0.128(P＜0.001).Conclusion Psychache fully mediates the relationship between peer victimization,school connectedness,and non-suicidal self-injury in adolescents,but it does not mediate the relationship between family functioning and adolescent non-suicidal self-injury.

Objective:To explore the relationship between anxiety symptoms and dispositional mindfulness and analyze the role of psychological resilience and interpersonal sensitivity in their relationship.Methods:A total of 478 college students were selected and assessed with the Depression Anxiety Stress Scale-Anxiety subscale(DASS-A),Mindful Attention Awareness Scale(MAAS),Brief Resilience Scale(BRS)and Symptom Checklist 90-Interpersonal Sensitivity Scale(IPS).SPSS macro program PROCESS3.5 was used to test the mediating role.Results:The DASS-A scores were negatively correlated with the MAAS and BRS scores(r=-0.38,-0.33,Ps＜0.01),and positively correlated with the IPS scores(r=0.40,P＜0.01).The MAAS scores were positively correlated with the BRS scores(r=0.40,P＜0.01),and negatively with the IPS scores(r=-0.39,P＜0.01).The BRS scores were negatively correlated with the IPS scores(r=-0.42,P＜0.01).Psychological resilience and interpersonal sensitivity acted as mediators and chain mediators between dispositional mindfulness and anxiety symptoms,with effect sizes of-0.06,-0.07,-0.03,accounting for 15.79％,18.42％,7.89％of the total effect,respectively.Conclusion:It suggests that anxiety symptoms are related to dispositional mindfulness in college students,and psychological resilience and interpersonal sensitivity play a chain mediating role between them.

Objective:To explore the relationship between anxiety symptoms and dispositional mindfulness and analyze the role of psychological resilience and interpersonal sensitivity in their relationship.Methods:A total of 478 college students were selected and assessed with the Depression Anxiety Stress Scale-Anxiety subscale(DASS-A),Mindful Attention Awareness Scale(MAAS),Brief Resilience Scale(BRS)and Symptom Checklist 90-Interpersonal Sensitivity Scale(IPS).SPSS macro program PROCESS3.5 was used to test the mediating role.Results:The DASS-A scores were negatively correlated with the MAAS and BRS scores(r=-0.38,-0.33,Ps＜0.01),and positively correlated with the IPS scores(r=0.40,P＜0.01).The MAAS scores were positively correlated with the BRS scores(r=0.40,P＜0.01),and negatively with the IPS scores(r=-0.39,P＜0.01).The BRS scores were negatively correlated with the IPS scores(r=-0.42,P＜0.01).Psychological resilience and interpersonal sensitivity acted as mediators and chain mediators between dispositional mindfulness and anxiety symptoms,with effect sizes of-0.06,-0.07,-0.03,accounting for 15.79％,18.42％,7.89％of the total effect,respectively.Conclusion:It suggests that anxiety symptoms are related to dispositional mindfulness in college students,and psychological resilience and interpersonal sensitivity play a chain mediating role between them.

Objective To explore the effects of family functioning,peer victimization and school connectedness on nonsuicidal self-injury in adolescents,as well as the mediating effect of psychache in these relationships.Methods A total of 525 junior high school students in Foshan City were surveyed using the brief Family Function Scale,Peer Victimization Scale,School Connectedness Scale,Psychache Scale,and Nonsuicidal Self-injury Scale.Descriptive statistics and correlation analyses were performed using SPSS 23.0,and mediation analysis was conducted using Mplus 7.4.Results ① Family functioning could directly predict adolescent non-suicidal self-injury(β=-0.20,P＜0.01),but did not exert an indirect effect through psychache(P=0.123).② Peer victimization could not directly predict adolescent non-suicidal self-injury(β=0.10,P=0.133),but had an indirect effect through psychache,with an indirect effect value of 0.142(P＜0.001).③ School connectedness could not directly predict adolescent non-suicidal self-injury(β=0.13,P=0.053),but had an indirect effect through psychache,with an indirect effect value of-0.128(P＜0.001).Conclusion Psychache fully mediates the relationship between peer victimization,school connectedness,and non-suicidal self-injury in adolescents,but it does not mediate the relationship between family functioning and adolescent non-suicidal self-injury.

Objective:To prepare a humanized monoclonal antibody against periostin and establish a stable cell line.Meth-ods:Based on anti-periostin mouse monoclonal antibody developed by our laboratory,total RNA was extracted,and variable region sequences were obtained by RT-PCR amplification of VH and VL genes.The mouse antibody CDR region was transplanted into the human antibody framework receptor region,and the gene was subcloned into the expression vector PATX-GS2,and stably transfected into CHO cells.Monoclonal cell lines were obtained by MSX pressure screening and limited dilution.Results:VH and VL genes were amplified by RT-PCR,and the sequence of the light and heavy chain variable region were determined.Antibody humanization were successfully stablished by CDR transplantation method a murine antibody to a human framework,and a eukaryotic expression plasmid was constructed,which was transfected into CHO cells for expression,and human anti-periostin antibody was successfully obtained.ELISA and Western blot results showed that the humanized antibody had good anti-periostin activities and binding affinity.Conclu-sion:In this study,anti-periostin humanized monoclonal antibody has been successfully prepared,which can specifically bind to peri-ostin proteins in vivo and have biological activity,providing scientific data for the precise treatment of retinal fibrosis,tissue and organ fibrosis,and malignant tumors.

Understanding the connection between brain and behavior in animals requires precise monitoring of their behaviors in three-dimensional (3-D) space. However, there is no available three-dimensional behavior capture system that focuses on rodents. Here, we present MouseVenue3D, an automated and low-cost system for the efficient capture of 3-D skeleton trajectories in markerless rodents. We improved the most time-consuming step in 3-D behavior capturing by developing an automatic calibration module. Then, we validated this process in behavior recognition tasks, and showed that 3-D behavioral data achieved higher accuracy than 2-D data. Subsequently, MouseVenue3D was combined with fast high-resolution miniature two-photon microscopy for synchronous neural recording and behavioral tracking in the freely-moving mouse. Finally, we successfully decoded spontaneous neuronal activity from the 3-D behavior of mice. Our findings reveal that subtle, spontaneous behavior modules are strongly correlated with spontaneous neuronal activity patterns.
Animals ; Behavior, Animal ; Brain/diagnostic imaging* ; Imaging, Three-Dimensional/methods* ; Mice ; Neuroimaging ; Rodentia

Objective:To investigate the effects of rapamycin (RAPA) on the cognitive function of lupus mice by regulating Cysteine rich 61 (Cyr61) and autophagy.Methods:MRL/lpr lupus mice were randomly divided into lupus group and rapamycin + lupus group, wild-type C57BL/6 mice were randomly divided into normal control group and rapamycin group with six mice in each group, RAPA + lupus group and rapamycin group were intraperitoneally injected with RAPA (2.0 mg/kg). The lupus group and the normal control group were injected with equal amounts of dimethyl sulfoxide (DMSO). Morris water maze was used to observe the cognitive function of mice. Western blotting was used to detect the expression of Cyr61, Programmed cell death-1 (Beclin-1), Microtubule associated protein 1 light chain3B (LC3B). Hematoxylin-eosin (HE) staining was used to observe the pathological changes in the hippocampus. The changes of neurons and bodies in hippocampus were observed by Nissl staining. The localization and expression of Cyr61 and LC3B in hippocampus were detected by immunofluorescence staining. One-way analysis of variance (ANOVA) was used between groups, and LSD-T test was used for pairwise comparison.Results:Western blotting results showed thatthe protein expression of Cyr61 (0.99±0.15) was significantly increased ( P=0.011), and the protein expression of Beclin-1 (0.64±0.04) and LC3B(0.54±0.05) was significantly decreased in lupus group ( P=0.025, P= 0.008) when compared with normal control group (0.73±0.08, 0.81±0.12, 0.80±0.03). The expressions of Cyr61 (0.75±0.05, 0.75±0.08), Beclin-1 (0.84±0.08, 0.92±0.04) and LC3B (0.93±0.16, 0.76±0.08) in rapamycin group and rapamycin + lupus group were not significantly changed ( P>0.05). Compared with rapamycin group, the protein expression of Cyr61 (0.99±0.15) was significantly increased ( P=0.016), and Beclin-1 (0.64±0.04), LC3B (0.54±0.05) was significantly decreased in lupus group ( P=0.013, P=0.001). The expressions of Cyr61 (0.75± 0.08), Beclin-1 (0.92±0.04) and LC3B (0.76±0.08) were not significantly changed in rapamycin+lupus group ( P=0.999, P=0.241, P=0.062). Compared with lupus group, the expression of Cyr61 (0.75±0.08) protein in rapamycin+lupus group was significantly decreased ( P=0.016), and the expression of Beclin-1 (0.92±0.04) and LC3B(0.76±0.08) protein were significantly increased ( P=0.002, P=0.017). Immunofluorescence results showed that Cyr61 and LC3B were mainly expressed in the cytoplasm of hippocampal neurons, and the quantitative detection results were consistent with western blot results, the differences were statistically significant ( P=0.025, P=0.032). HE staining showed that the levels and number of cells in the hippocampus of mice with lupus were reduced, and the arrangement was sparse, and the nuclei were hyperchromatic, showing nuclear pyknosis and migration. The results of Nissl staining showed that there were relatively fewer Nissl bodies, loose arrangement of neurons and vacuolar areas in some cells, which were improved after RAPA treatment in lupus mice. Conclusion:RAPA can protect the cognitive function of lupus mice by inhibiting the expression of Cyr61 in hippocampus and promoting autophagy.