Objective:To investigate the effect of curcumin on phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway on the autophagy of Bacille Calmette-Guérin (BCG)-infected macrophages.Methods:The infection model was established by infecting THP-1-derived macrophages with BCG. Five groups were involved in this study, which were control group, BCG group, BCG+ curcumin group, BCG+ curcumin+ IGF-1(PI3K agonist) group, and BCG+ curcumin+ LY294002 (PI3K inhibitor) group. The fluorescence intensity of autophagosomes was observed under fluorescence microscope using the fluorescent dye monodansylcadaverine (MDC staining). The expression of PI3K, Akt, mTOR, phospho-PI3K (p-PI3K), phospho-Akt (p-Akt), phospho-mTOR (p-mTOR), microtubule-associated protein 1 light chain 3-Ⅱ (LC3-Ⅱ), and Beclin-1 at protein level were detected by Western blot. Colony forming unit was used to detect macrophage load. Multiple independent, normal, and homogeneous data were compared using one-way analysis of variance, and pairwise comparisons were conducted using LSD test.Results:BCG infection significantly decreased the fluorescence intensity of autophagosomes, and the expression of autophagy marker proteins LC3-Ⅱ and Beclin-1 ( P<0.05), but increased the expression of p-PI3K, p-Akt, and p-mTOR ( P<0.05). Curcumin increased the fluorescence intensity of autophagosomes and enhanced the expression of LC3-Ⅱ and Beclin-1 proteins in a concentration-dependent manner ( P<0.05). Besides, curcumin inhibited the expression of p-PI3K, p-Akt, and p-mTOR ( P<0.05). The PI3K agonist IGF-1 reversed the above effects of curcumin. Compared with the BCG+ curcumin group, the fluorescence intensity of autophagosomes and the expression of LC3-Ⅱ and Beclin-1 proteins were further increased ( P<0.05), while the expression of p-PI3K, p-Akt and p-mTOR was further decreased ( P<0.05) in the BCG+ curcumin+ LY294002 group. Compared with the BCG group, the bacterial loads in the BCG+ curcumin group and the BCG+ curcumin+ LY294002 group decreased significantly ( P<0.05), while the bacterial load in the BCG+ curcumin+ IGF-1 group increased significantly ( P<0.05). Conclusions:Curcumin can promote the autophagy of BCG-infected macrophages, which contributes to the clearance of Mycobacterium tuberculosis by macrophages. Part of the mechanism may be related to the inhibition of PI3K/Akt/mTOR pathway.

Objective To explore the molecular mechanism by which curcumin enhances the anti-tuberculosis function of macrophages through immune metabolic regulation mediated by liver X receptor α(LXRα)/ABCA1.Methods A model was established by infecting THP-1-derived macrophages with attenuated strain of Mycobacterium bovis(M.bovis).The control group,curcumin group,M.pavis group,M.pavis+LXRα agonist(T0901317)group,M.pavis+LXRα inhibitor(GSK2033)group,M.pavis+curcumin group,M.pavis+curcumin+GSK2033 group and M.pavis+curcumin+T0901317 group were set up.The protein and gene expressions of LXRα/ABCA1 were detected by Western blotting and real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).The accumulation of lipid droplets was analyzed by Oil Red O staining and micro-assay.The lipid content of the supernatant was determined by a biochemical analyzer,and cell proliferation was assessed by the MTT method.Bacterial clearance capacity was evaluated by measuring intracellular bacterial load.Results Curcumin significantly upregulated the protein and gene expression of LXRα/ABCA1 in M.Bovis-infected macrophages,reduced intracellular lipid accumulation and promoted lipid efflux,while enhancing cell proliferation and reducing intracellular bacterial load(P<0.05,P<0.01).LXRα inhibitors could reverse the effect of curcumin,while agonists synergistically enhanced its effect.Correlation analysis showed that the expression of LXRα/ABCA1 in cells was negatively correlated with the intracellular bacterial load,while the lipid level was positively correlated with the intracellular bacterial load(P<0.01).Conclusion Curcumin activates the LXRα/ABCA1 pathway,coordinates the metabolic remodeling of macrophages and the enhancement of immune function,and forms a synergistic effect against tuberculosis,providing an experimental basis for the development of a novel host-directed treatment strategy for tuberculosis based on immune-metabolic regulation.

Objective To explore the molecular mechanism by which curcumin enhances the anti-tuberculosis function of macrophages through immune metabolic regulation mediated by liver X receptor α(LXRα)/ABCA1.Methods A model was established by infecting THP-1-derived macrophages with attenuated strain of Mycobacterium bovis(M.bovis).The control group,curcumin group,M.pavis group,M.pavis+LXRα agonist(T0901317)group,M.pavis+LXRα inhibitor(GSK2033)group,M.pavis+curcumin group,M.pavis+curcumin+GSK2033 group and M.pavis+curcumin+T0901317 group were set up.The protein and gene expressions of LXRα/ABCA1 were detected by Western blotting and real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).The accumulation of lipid droplets was analyzed by Oil Red O staining and micro-assay.The lipid content of the supernatant was determined by a biochemical analyzer,and cell proliferation was assessed by the MTT method.Bacterial clearance capacity was evaluated by measuring intracellular bacterial load.Results Curcumin significantly upregulated the protein and gene expression of LXRα/ABCA1 in M.Bovis-infected macrophages,reduced intracellular lipid accumulation and promoted lipid efflux,while enhancing cell proliferation and reducing intracellular bacterial load(P<0.05,P<0.01).LXRα inhibitors could reverse the effect of curcumin,while agonists synergistically enhanced its effect.Correlation analysis showed that the expression of LXRα/ABCA1 in cells was negatively correlated with the intracellular bacterial load,while the lipid level was positively correlated with the intracellular bacterial load(P<0.01).Conclusion Curcumin activates the LXRα/ABCA1 pathway,coordinates the metabolic remodeling of macrophages and the enhancement of immune function,and forms a synergistic effect against tuberculosis,providing an experimental basis for the development of a novel host-directed treatment strategy for tuberculosis based on immune-metabolic regulation.

Objective:To investigate the effect of curcumin on phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway on the autophagy of Bacille Calmette-Guérin (BCG)-infected macrophages.Methods:The infection model was established by infecting THP-1-derived macrophages with BCG. Five groups were involved in this study, which were control group, BCG group, BCG+ curcumin group, BCG+ curcumin+ IGF-1(PI3K agonist) group, and BCG+ curcumin+ LY294002 (PI3K inhibitor) group. The fluorescence intensity of autophagosomes was observed under fluorescence microscope using the fluorescent dye monodansylcadaverine (MDC staining). The expression of PI3K, Akt, mTOR, phospho-PI3K (p-PI3K), phospho-Akt (p-Akt), phospho-mTOR (p-mTOR), microtubule-associated protein 1 light chain 3-Ⅱ (LC3-Ⅱ), and Beclin-1 at protein level were detected by Western blot. Colony forming unit was used to detect macrophage load. Multiple independent, normal, and homogeneous data were compared using one-way analysis of variance, and pairwise comparisons were conducted using LSD test.Results:BCG infection significantly decreased the fluorescence intensity of autophagosomes, and the expression of autophagy marker proteins LC3-Ⅱ and Beclin-1 ( P<0.05), but increased the expression of p-PI3K, p-Akt, and p-mTOR ( P<0.05). Curcumin increased the fluorescence intensity of autophagosomes and enhanced the expression of LC3-Ⅱ and Beclin-1 proteins in a concentration-dependent manner ( P<0.05). Besides, curcumin inhibited the expression of p-PI3K, p-Akt, and p-mTOR ( P<0.05). The PI3K agonist IGF-1 reversed the above effects of curcumin. Compared with the BCG+ curcumin group, the fluorescence intensity of autophagosomes and the expression of LC3-Ⅱ and Beclin-1 proteins were further increased ( P<0.05), while the expression of p-PI3K, p-Akt and p-mTOR was further decreased ( P<0.05) in the BCG+ curcumin+ LY294002 group. Compared with the BCG group, the bacterial loads in the BCG+ curcumin group and the BCG+ curcumin+ LY294002 group decreased significantly ( P<0.05), while the bacterial load in the BCG+ curcumin+ IGF-1 group increased significantly ( P<0.05). Conclusions:Curcumin can promote the autophagy of BCG-infected macrophages, which contributes to the clearance of Mycobacterium tuberculosis by macrophages. Part of the mechanism may be related to the inhibition of PI3K/Akt/mTOR pathway.

Objective Network pharmacology,Molecular docking and Label-free DIA quantitative phosphoproteomics were used to reveal the potential mechanism of Peiminine against colon cancer.Methods 1The target of peiminine was obtained by SwissTargetPrediction,TargetNet and pharmmapper,and the target of colon cancer was obtained by DisGeNET,GeneCards and OMIM.Then the intersection target of Peiminine and Colon cancer was obtained by Venny2.1.0 online platform.Then,String database and Cytoscape3.8.2 software were used to map the PPI network of intersection targets,and the main targets of Peiminine against Colon cancer were obtained.GO analysis and KEGG pathway analysis were carried out through David database and Weisenxin visual cloud platform.② MOE(molecular operating environment)software was used to perform molecular docking of peiminine and the main target.③ Label-free DIA quantitative phosphoproteomics was used to detect and analyze the biological function of DT group(DT1-DT3)treated with Peiminine and control group(NC1-NC3).Results ① There were 275 intersection targets between peiminine and colon cancer.Molecular docking showed that peiminine could stably dock and interact with the protein structures of AKT1,EGFR,HSP90AA1 and SRC:Peiminine interacted with the amino acid residues of AKT1 mainly through hydrogen bonding.Peiminine interacted with amino acid residues of EGFR,HSP90AA1 and SRC mainly through ionic bond and hydrogen bonding.② Phosphoproteomics analysis showed that:Compared with the NC group,880 phosphorylated modification sites were significantly up-regulated in the DT group(including S124 and S126 sites of AKT1 and T648 and S643 sites of EGFR),and 425 phosphorylated modification sites were significantly down-regulated in the DT group(including T317 sites of HSP90AA1).③ Comparing the results of network pharmacology and phosphoproteomics analysis,it was found that:The main targets of Peiminine against Colon cancer are AKT1,EGFR and HSP90AA1.It promotes apoptosis of colon cancer cells by regulating 17 pathways including AMPK signaling pathway,mTOR signaling pathway and Choline metabolism in cancer.Conclusion This study revealed the potential mechanism of peiminine in the treatment of colon cancer with multiple targets and multiple pathways,and provided a certain direction and reference for subsequent research.

Objective Network pharmacology,Molecular docking and Label-free DIA quantitative phosphoproteomics were used to reveal the potential mechanism of Peiminine against colon cancer.Methods 1The target of peiminine was obtained by SwissTargetPrediction,TargetNet and pharmmapper,and the target of colon cancer was obtained by DisGeNET,GeneCards and OMIM.Then the intersection target of Peiminine and Colon cancer was obtained by Venny2.1.0 online platform.Then,String database and Cytoscape3.8.2 software were used to map the PPI network of intersection targets,and the main targets of Peiminine against Colon cancer were obtained.GO analysis and KEGG pathway analysis were carried out through David database and Weisenxin visual cloud platform.② MOE(molecular operating environment)software was used to perform molecular docking of peiminine and the main target.③ Label-free DIA quantitative phosphoproteomics was used to detect and analyze the biological function of DT group(DT1-DT3)treated with Peiminine and control group(NC1-NC3).Results ① There were 275 intersection targets between peiminine and colon cancer.Molecular docking showed that peiminine could stably dock and interact with the protein structures of AKT1,EGFR,HSP90AA1 and SRC:Peiminine interacted with the amino acid residues of AKT1 mainly through hydrogen bonding.Peiminine interacted with amino acid residues of EGFR,HSP90AA1 and SRC mainly through ionic bond and hydrogen bonding.② Phosphoproteomics analysis showed that:Compared with the NC group,880 phosphorylated modification sites were significantly up-regulated in the DT group(including S124 and S126 sites of AKT1 and T648 and S643 sites of EGFR),and 425 phosphorylated modification sites were significantly down-regulated in the DT group(including T317 sites of HSP90AA1).③ Comparing the results of network pharmacology and phosphoproteomics analysis,it was found that:The main targets of Peiminine against Colon cancer are AKT1,EGFR and HSP90AA1.It promotes apoptosis of colon cancer cells by regulating 17 pathways including AMPK signaling pathway,mTOR signaling pathway and Choline metabolism in cancer.Conclusion This study revealed the potential mechanism of peiminine in the treatment of colon cancer with multiple targets and multiple pathways,and provided a certain direction and reference for subsequent research.

Objective:To evaluate the influencing factors for venous thromboembolism (VTE) in patients undergoing neurosurgery, so as to provide a reference for the establishment of preventive strategies for VTE in patients undergoing neurosurgery.Methods:Literatures related to the influencing factors of VTE in patients undergoing neurosurgery were searched systematically in PubMed, Embase, Cochrane Library, CINAHL, MEDLINE, CNKI, Wanfang Database, VIP, China Biomedical Literature Database (CBM) . The retrieval time limit was from the establishment of database to December 31, 2021. Literature quality was evaluated by 2 researchers using the JBI Evidence-based Health Care Centre Evaluation tool in Australia. RevMan 5.4 software was used for Meta-analysis.Results:A total of 23 articles were included. The results of Meta-analysis showed that age>45 years ( OR=2.27, 95% CI: 1.44-3.58, P<0.01) , male ( OR=0.88, 95% CI: 0.79-0.98, P=0.03) , high BMI ( OR=1.68, 95% CI: 0.12-3.23, P=0.03) , hypertension ( OR=2.27, 95% CI: 1.66-3.11, P<0.01) , smoking history ( OR=1.51, 95% CI: 1.14-2.02, P=0.005) , history of previous thrombosis ( OR=6.21, 95% CI: 1.50-25.70, P=0.01) , Caucasian ( OR=1.55, 95% CI: 1.26-1.89, P<0.01) , Hispanic ( OR=0.74, 95% CI: 0.58-0.94, P=0.01) , Asian ( OR=0.37, 95% CI: 0.22-0.62, P=0.01) , lower extremity weakness or paralysis ( OR=6.14, 95% CI: 2.24-16.81, P=0.000 4) , high glioma grading ( OR=1.86, 95% CI: 1.01-3.41, P=0.04) , nerve sheath tumor ( OR=7.87, 95% CI: 1.07-57.86, P=0.04) , tumor brain metastasis ( OR=1.92, 95% CI: 1. 38-2.68, P<0.01) , intraoperative supine position ( OR=1.65, 95% CI: 1.07-2.55, P=0.02) , use of dehydrating agents ( OR=2.85, 95% CI: 1.33-6.09, P<0.01) , infection ( OR=16.23, 95% CI: 7.84-33.64, P<0.01) , central venous line placement ( OR=9.69, 95% CI: 4.80-19.58, P<0.01) were the risk factors of VTE in neurosurgery patients. Conclusions:The VTE in neurosurgery patients is affected by many factors. Medical and nursing staff should identify risk factors timely, and take targeted preventive measures and reduce the incidence of VTE.

Alzheimer' s disease (AD) is a neurodegenerative disease that gradually impairs cognitive functions. Recently, there has been a conceptual shift toward AD to view the disease as a continuum. Since AD is currently incurable, effective intervention to delay or prevent pathological cognitive decline may best target the early stages of symptomatic disease, such as subjective cognitive decline (SCD), in which cognitive function remains relatively intact. Diagnostic methods for identifying AD, such as cerebrospinal fluid biomarkers and positron emission tomography, are invasive and expensive. Therefore, it is imperative to develop blood biomarkers that are sensitive, less invasive, easier to access, and more cost effective for AD diagnosis. This review aimed to summarize the current data on whether individuals with SCD differ reliably and effectively in subjective and objective performances compared to cognitively normal elderly individuals, and to find one or more convenient and accessible blood biomarkers so that researchers can identify SCD patients with preclinical AD in the population as soon as possible. Owing to the heterogeneity and complicated pathogenesis of AD, it is difficult to make reliable diagnoses using only a single blood marker. This review provides an overview of the progress achieved to date with the use of SCD blood biomarkers in patients with preclinical AD, highlighting the key areas of application and current challenges.

Objective:To investigate the inhibitory effect of curcumin on oxidative stress in BCG-infected macrophages based on the Nrf2 pathway.Methods:THP-1-derived macrophages were infected.The experiment was divided into control group,BCG group,BCG+curcumin group and BCG+curcumin+ML385 group.Cellular ROS fluorescence intensity were observed under a fluores-cence microscope;Glutathione(GSH)levels were measured by Colorimetry;Western blot was used to detect the protein expressions of Nrf2,HO-1 and NQO1;MTT was used to detect the proliferation rate of macrophages.Results:BCG infection significantly enhanced ROS fluorescence intensity,reduced cell GSH content(P<0.01),inhibited protein expressions of Nrf2,HO-1 and NQO1,at the same time inhibited cell proliferation(P<0.01);curcumin significantly weakened ROS fluorescence intensity,increased GSH level(P<0.05),promoted Nrf2,HO-1 and NQO1 protein expressions and cell proliferation(P<0.01);Nrf2 inhibitor ML385 reversed the effect of curcumin.Conclusion:Curcumin can alleviate BCG-induced oxidative stress in macrophages by increasing the expression of Nrf2 and inducing the transcription of downstream antioxidant molecules.

Objective:To design and construct a bone nonunion organoid on chip and explore the mechanism of aseptic bone nonunion.Methods:First a semi-open microfluidic chip was designed, on which human bone marrow mesenchymal stromal cells (BMSC), human fetal lung fibroblast 1, (HFL1) and human umbilical vein endothelial cells (HUVEC) were co-cultured, and a three-dimensional organ on chip system was established. Different proportions of HFL1 and HUVEC were co-cultured with BMSC, which were divided into the control group (HFL1∶HUVEC=1∶1), the fibrosis group (HFL1∶HUVEC=3∶1) and the vascularization group (HFL1∶HUVEC=1∶3). The osteogenic differentiation of BMSC was observed by alkaline phosphatase (ALP) and Alizarin red staining. The transcription level of osteogenic marker genes SP7, RUNX2, ALPL, and BGLAP, and vascularization related genes KDR and VWF were analyzed by qPCR. The expression levels of RUNX2 and ALP were determined by Western Blot. Results:In the co-culture system of BMSCs, HFL1, and HUVECs, BMSCs exhibited normal growth and apparent biomineralization behavior. Endothelial cells were capable of forming structured vascular networks, confirming the successful establishment of the system. Compared to the baseline group, the fibrotic group showed no significant decrease in BMSC osteogenic differentiation. The relative expression levels of the mineralization marker genes ALPL and BGLAP were 0.55±0.19 ( P<0.001) and 0.42±0.27 ( P<0.001), respectively. Vascularization genes KDR and VWF were downregulated, with relative expression levels of 0.49±0.17 ( P<0.001) and 0.49±0.21 ( P<0.001). In contrast, in the vascularized group, BMSC osteogenic differentiation genes SP7, RUNX2, ALPL, and BGLAP were upregulated, with relative expression levels of 2.91±0.52 ( P<0.001), 3.83±1.87 ( P<0.001), 3.22±1.29 ( P<0.001), and 5.21±1.46 ( P<0.001), respectively. Vascularization genes KDR and VWF were also upregulated, with relative expressions of 8.24±2.84 ( P<0.001) and 5.32±1.67 ( P<0.001). Western blot results indicated increased expression of RUNX2 and ALP in the vascularized group and decreased expression in the fibrotic group. Conclusion:The bone nonunion organoid on chip could partially simulate the local microenvironment of bone nonunion. Fibrosis may lead to a significant decrease in bone formation ability and vascularization level, which might be an important reason for the occurrence of aseptic bone nonunion.