Objective To investigate the biological behavior of EMP1 in pancreatic cancer cells and the molecular mechanism of EMP1 in promoting tumor progression.Methods A stable EMP1 knockdown cell line was obtained by lentivirus transfection.The effect of EMP1 on the proliferation of cancer cells was determined by CCK-8 and clonal formation assay.The effect of EMP1 on the migration and invasion of cancer cells was detected by scratch test and Transwell test.The influence of EMP1 on downstream signaling pathways was investigated by Western blot.Results The results of qRT-PCR and Western blot showed that EMP1 was highly expressed in pancreatic cancer cells.The results of CCK-8,colony formation,scratch,and Transwell assays indicated that EMP1 promoted the proliferation,migration,and invasion of pancreatic cancer cells.Western blot results revealed that EMP1 might promote tumor progression through the PI3K/AKT signaling pathway.Conclusion This study suggested that EMP1 may activate the PI3K/AKT signaling pathway to promote the proliferation,migration,and invasion of pancreatic cancer cells,thereby positively regulating tumor progression.

Depression is a widespread psychiatric disorder with complex pathogenesis and unsatisfactory therapeutic effects. As a native flavonoid, Isorhamnetin (ISO) has been deemed to exert neuroprotective effects by antioxidation and regulation of immunity. However, no reports of anti-depressed effect of ISO have yet been found. The present study was conducted to clarify the mechanism basis of anti-depressed effect of ISO utilizing behavioral, biochemical, molecular approaches in vitro and in vivo and bio-informatics analysis. The effects of ISO on depressed mice was investigated through the SPT and FST, and the lesions were examined by H&E staining. Besides, the inflammatory factor and indicator in kynurenine pathway were assessed through detection kits, and the microbiota were checked by 16sRNA. Molecular docking study was performed to investigate the target of ISO. Additionally, Western blot was used to test the activation of PI3K/AKT signaling pathway. The results indicated that ISO could enhance the sugar water preference of mice in SPT and reduce immobility time in FST. Further more, ISO suppressed peripheral and central inflammation, regulated the changes in kynurenine pathway and gut microbiota, inhibited activation of PI3K/AKT pathway, and presented good binding patterns with target proteins on PI3K/AKT signaling pathway. Collectively, these findings demonstrate that ISO alleviated depression-like behaviour by normalizing inflammation-induced dysregulation of the crosstalk between KP and gut microbiota disorder through regulated PI3K/AKT/NF-κB pathway.

Depression is a widespread psychiatric disorder with complex pathogenesis and unsatisfactory therapeutic effects. As a native flavonoid, Isorhamnetin (ISO) has been deemed to exert neuroprotective effects by antioxidation and regulation of immunity. However, no reports of anti-depressed effect of ISO have yet been found. The present study was conducted to clarify the mechanism basis of anti-depressed effect of ISO utilizing behavioral, biochemical, molecular approaches in vitro and in vivo and bio-informatics analysis. The effects of ISO on depressed mice was investigated through the SPT and FST, and the lesions were examined by H&E staining. Besides, the inflammatory factor and indicator in kynurenine pathway were assessed through detection kits, and the microbiota were checked by 16sRNA. Molecular docking study was performed to investigate the target of ISO. Additionally, Western blot was used to test the activation of PI3K/AKT signaling pathway. The results indicated that ISO could enhance the sugar water preference of mice in SPT and reduce immobility time in FST. Further more, ISO suppressed peripheral and central inflammation, regulated the changes in kynurenine pathway and gut microbiota, inhibited activation of PI3K/AKT pathway, and presented good binding patterns with target proteins on PI3K/AKT signaling pathway. Collectively, these findings demonstrate that ISO alleviated depression-like behaviour by normalizing inflammation-induced dysregulation of the crosstalk between KP and gut microbiota disorder through regulated PI3K/AKT/NF-κB pathway.

Depression is a widespread psychiatric disorder with complex pathogenesis and unsatisfactory therapeutic effects. As a native flavonoid, Isorhamnetin (ISO) has been deemed to exert neuroprotective effects by antioxidation and regulation of immunity. However, no reports of anti-depressed effect of ISO have yet been found. The present study was conducted to clarify the mechanism basis of anti-depressed effect of ISO utilizing behavioral, biochemical, molecular approaches in vitro and in vivo and bio-informatics analysis. The effects of ISO on depressed mice was investigated through the SPT and FST, and the lesions were examined by H&E staining. Besides, the inflammatory factor and indicator in kynurenine pathway were assessed through detection kits, and the microbiota were checked by 16sRNA. Molecular docking study was performed to investigate the target of ISO. Additionally, Western blot was used to test the activation of PI3K/AKT signaling pathway. The results indicated that ISO could enhance the sugar water preference of mice in SPT and reduce immobility time in FST. Further more, ISO suppressed peripheral and central inflammation, regulated the changes in kynurenine pathway and gut microbiota, inhibited activation of PI3K/AKT pathway, and presented good binding patterns with target proteins on PI3K/AKT signaling pathway. Collectively, these findings demonstrate that ISO alleviated depression-like behaviour by normalizing inflammation-induced dysregulation of the crosstalk between KP and gut microbiota disorder through regulated PI3K/AKT/NF-κB pathway.

Background and purpose:Prostate cancer and breast cancer are highly prevalent malignant tumors,and there occurrence and development are related to the tumor suppressor genes phosphatase and tensin homolog deleted on chremosome ten(Pten)and the transformation related protein 53 gene(Trp53).The loss of function of Trp53 is closely related.The simultaneous loss of the two can accelerate the malignant progression of tumors and induce therapeutic resistance.The gene-edited spontaneous tumor model of mice based on the Cre-loxP system is a key tool for studying the mechanism of cancer.Studies have shown that prostate-specific promoter(probasin,Pbsn)-driven iCre recombinase(Pbsn-iCre)can induce spontaneous prostate cancer in male mice,but its role in female breast cancer and transgender expression characteristics have not yet been clarified.In this study,we constructed Ptenfl/fl;Trp53fl/fl;Pbsn-iCre+transgenic mouse model which was designed to explore its spontaneous tumor phenotype in prostate cancer and breast cancer,and to verify the expression characteristics of Pbsn in breast tissue.Methods:The Ptenfl/fl;Trp53fl/fl;Pbsn-iCre+mouse model was established using Cre-loxP system by hybridization and continuous backcross screening with Ptenfl/fl mouse,Trp53fl/flmouse,and Pbsn-iCre+mouse(Ethical No.:FUSCC-IACUC-2025115).Pten,Trp53 and Pbsn-iCre genotypes were verified by polymerase chain reaction and agarose gel electrophoresis.The incidence of tumor in transgenic mice was monitored,and the histopathological characteristics of tumor were evaluated by hematoxylin-eosin staining.The protein levels of Pten and p53 in prostate and breast tumor tissues were analyzed by immunohistochemistry,and the distributions of Pbsn in breast,prostate,ovary,heart,liver and kidney were detected.Results:Ptenfl/fl;Trp53fl/fl;Pbsn-iCre+male mouse developed spontaneous prostate tumor at age of 5 month,and female mouse developed spontaneous breast tumor at age of 6 months.The pathological manifestations of prostate cancer were invasive acinar adenocarcinoma structure with glandular structure disorder and basement membrane destruction.The pathological manifestations of breast cancer were invasive ductal carcinoma with ductal epithelial dysplasia and interstitial lymphocyte infiltration.Immunohistochemistry confirmed the complete deletion of Pten and p53 proteins in prostate and breast tumor tissues,which verified the prostate and mammary gland specific gene knockout effect.Immunohistochemistry also confirmed that Pbsn protein was specifically expressed in prostate acinar epithelial cells,ovarian tissue,and mammary duct epithelial cells,but not in heart,liver and kidney.Conclusion:Pbsn-iCre is functionally expressed in female mammary glands,and the simultaneous loss of Pten/Trp53 induced by Pbsn-iCre may drive the development of prostate cancer in male and breast cancer in female mouse.

Objective To investigate the biological behavior of EMP1 in pancreatic cancer cells and the molecular mechanism of EMP1 in promoting tumor progression.Methods A stable EMP1 knockdown cell line was obtained by lentivirus transfection.The effect of EMP1 on the proliferation of cancer cells was determined by CCK-8 and clonal formation assay.The effect of EMP1 on the migration and invasion of cancer cells was detected by scratch test and Transwell test.The influence of EMP1 on downstream signaling pathways was investigated by Western blot.Results The results of qRT-PCR and Western blot showed that EMP1 was highly expressed in pancreatic cancer cells.The results of CCK-8,colony formation,scratch,and Transwell assays indicated that EMP1 promoted the proliferation,migration,and invasion of pancreatic cancer cells.Western blot results revealed that EMP1 might promote tumor progression through the PI3K/AKT signaling pathway.Conclusion This study suggested that EMP1 may activate the PI3K/AKT signaling pathway to promote the proliferation,migration,and invasion of pancreatic cancer cells,thereby positively regulating tumor progression.

Background and purpose:Prostate cancer and breast cancer are highly prevalent malignant tumors,and there occurrence and development are related to the tumor suppressor genes phosphatase and tensin homolog deleted on chremosome ten(Pten)and the transformation related protein 53 gene(Trp53).The loss of function of Trp53 is closely related.The simultaneous loss of the two can accelerate the malignant progression of tumors and induce therapeutic resistance.The gene-edited spontaneous tumor model of mice based on the Cre-loxP system is a key tool for studying the mechanism of cancer.Studies have shown that prostate-specific promoter(probasin,Pbsn)-driven iCre recombinase(Pbsn-iCre)can induce spontaneous prostate cancer in male mice,but its role in female breast cancer and transgender expression characteristics have not yet been clarified.In this study,we constructed Ptenfl/fl;Trp53fl/fl;Pbsn-iCre+transgenic mouse model which was designed to explore its spontaneous tumor phenotype in prostate cancer and breast cancer,and to verify the expression characteristics of Pbsn in breast tissue.Methods:The Ptenfl/fl;Trp53fl/fl;Pbsn-iCre+mouse model was established using Cre-loxP system by hybridization and continuous backcross screening with Ptenfl/fl mouse,Trp53fl/flmouse,and Pbsn-iCre+mouse(Ethical No.:FUSCC-IACUC-2025115).Pten,Trp53 and Pbsn-iCre genotypes were verified by polymerase chain reaction and agarose gel electrophoresis.The incidence of tumor in transgenic mice was monitored,and the histopathological characteristics of tumor were evaluated by hematoxylin-eosin staining.The protein levels of Pten and p53 in prostate and breast tumor tissues were analyzed by immunohistochemistry,and the distributions of Pbsn in breast,prostate,ovary,heart,liver and kidney were detected.Results:Ptenfl/fl;Trp53fl/fl;Pbsn-iCre+male mouse developed spontaneous prostate tumor at age of 5 month,and female mouse developed spontaneous breast tumor at age of 6 months.The pathological manifestations of prostate cancer were invasive acinar adenocarcinoma structure with glandular structure disorder and basement membrane destruction.The pathological manifestations of breast cancer were invasive ductal carcinoma with ductal epithelial dysplasia and interstitial lymphocyte infiltration.Immunohistochemistry confirmed the complete deletion of Pten and p53 proteins in prostate and breast tumor tissues,which verified the prostate and mammary gland specific gene knockout effect.Immunohistochemistry also confirmed that Pbsn protein was specifically expressed in prostate acinar epithelial cells,ovarian tissue,and mammary duct epithelial cells,but not in heart,liver and kidney.Conclusion:Pbsn-iCre is functionally expressed in female mammary glands,and the simultaneous loss of Pten/Trp53 induced by Pbsn-iCre may drive the development of prostate cancer in male and breast cancer in female mouse.

OBJECTIVES: To explore the role of the cGAS-STING signaling pathway in the therapeutic mechanism of Liangxue Jiedu Huayu Formula (LXJDHYF) for acute-on-chronic liver failure (ACLF) in mice. METHODS: Thirty C57BL/6 mice were randomly divided into blank control group, model group, low- and high-dose LXJDHYF groups, and H151 (a specific cGAS-STING pathway inhibitor) group (n=6). In all but the control group, the mice were treated with CCl₄ to induce liver cirrhosis followed by intraperitoneal injections of lipopolysaccharide and D-amino galactose to establish mouse models of ACLF. After the treatments, the mouse livers were collected for HE and TUNEL staining, and serum levels of ALT, AST and TBil were determined. In bone marrow-derived macrophages (BMDMs) and liver tissues of ACLF mice, the expressions of cGAS-STING signaling pathway-related mRNAs including IFN‑β, ISG15, IL-6 and TNF-α were determined with RT-qPCR, and the phosphorylation levels of IRF3 and STING proteins were investigated using Western blotting. RESULTS: Compared with the mice in the model group, the LXJDHYF-treated mice exhibited milder hepatocyte necrosis and inflammatory cell infiltration in the liver with significantly reduced hepatocyte apoptosis. LXJDHYF treatment also significantly lowered serum levels of ALT, AST, TBil, IL-6 and TNF-α in ACLF mice and effectively suppressed the expressions of cGAS-STING signaling pathway-related mRNA in both the BMDMs and the liver tissues and the phosphorylation of IRF3 and STING proteins in the BMDMs. CONCLUSIONS LXJDHYF can significantly improve liver function and attenuate inflammation in ACLF mice possibly by inhibiting excessive activation of the cGAS-STING signaling pathway.
Animals ; Signal Transduction/drug effects* ; Mice ; Nucleotidyltransferases/metabolism* ; Mice, Inbred C57BL ; Acute-On-Chronic Liver Failure/etiology* ; Membrane Proteins/metabolism* ; Drugs, Chinese Herbal/therapeutic use* ; Liver/metabolism* ; Disease Models, Animal ; Interferon Regulatory Factor-3/metabolism* ; Interleukin-6/metabolism* ; Male

Objective To explore the role of the cGAS-STING signaling pathway in the therapeutic mechanism of Liangxue Jiedu Huayu Formula(LXJDHYF)for acute-on-chronic liver failure(ACLF)in mice.Methods Thirty C57BL/6 mice were randomly divided into blank control group,model group,low-and high-dose LXJDHYF groups,and H151(a specific cGAS-STING pathway inhibitor)group(n=6).In all but the control group,the mice were treated with CC14 to induce liver cirrhosis followed by intraperitoneal injections of lipopolysaccharide and D-amino galactose to establish mouse models of ACLF.After the treatments,the mouse livers were collected for HE and TUNEL staining,and serum levels of ALT,AST and TBil were determined.In bone marrow-derived macrophages(BMDMs)and liver tissues of ACLF mice,the expressions of cGAS-STING signaling pathway-related mRNAs including IFN-β,ISG15,IL-6 and TNF-α were determined with RT-qPCR,and the phosphorylation levels of IRF3 and STING proteins were investigated using Western blotting.Results Compared with the mice in the model group,the LXJDHYF-treated mice exhibited milder hepatocyte necrosis and inflammatory cell infiltration in the liver with significantly reduced hepatocyte apoptosis.LXJDHYF treatment also significantly lowered serum levels of ALT,AST,TBil,IL-6 and TNF-α in ACLF mice and effectively suppressed the expressions of cGAS-STING signaling pathway-related mRNA in both the BMDMs and the liver tissues and the phosphorylation of IRF3 and STING proteins in the BMDMs.Conclusion LXJDHYF can significantly improve liver function and attenuate inflammation in ACLF mice possibly by inhibiting excessive activation of the cGAS-STING signaling pathway.

Spinal surgical site infection (SSI), especially deep SSI after internal fixation is difficult in treatment, with long course of disease and poor prognosis. At present, there are many controversies in the diagnosis and treatment of spinal SSI, with unsatisfactory overall efficacy of its diagnosis and treatment. Besides, no diagnosis and treatment guideline based on evidence-based medicine has been in existence. To this end, the Spinal Infection Group of the Orthopedic Branch of the Chinese Medical Doctor Association and the Spinal Infection Group of the Spinal Surgery Branch of the Chinese Rehabilitation Medicine Association jointly organized relevant experts to formulate Evidence-based clinical guideline for the diagnosis and treatment of surgical site infection in spinal trauma ( version 2024) based on an evidence-based approach. A total of 10 recommendations were proposed on the diagnosis and treatment of spinal SSI, so as to provide a clinical reference for the diagnosis and treatment of spinal SSI.