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: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 investigate the role of peroxisome proliferator-activated receptor γ (PPARγ)/CD36 signaling pathway in macrophage lipid metabolism after Mycobacterium tuberculosis ( Mtb) infection. Methods:THP-1-derived macrophages were infected with Mtb. Four groups were included in this study, which were control group, Mtb infection group, Mtb+ rosiglitazone (ROZ, PPARγ agonist) group and Mtb+ GW9662 (PPARγ antagonist) group. Western blot and RT-PCR were used to detect the expression of PPARγ in macrophages at protein and mRNA levels, respectively. The lipids in cells were detected by oil red O staining. The concentrations of total cholesterol (TC), triglycerides (TG), low density lipoprotein (LDL-C) and high density lipoprotein (HDL-C) in the supernatant of cell culture were detected by automatic biochemical analyzer. The expression of CD36 was detected by immunohistochemistry. CCK-8 was used to detect the proliferation rate of macrophages. Results:Mtb infection significantly increased the expression of PPARγ in macrophages ( P<0.001), promoted intracellular lipid aggregation and CD36 expression and decreased the levels of TC, TG, LDL-C and HDL-C in the supernatant of cell culture ( P<0.001) and cell proliferation rate ( P<0.001). PPARγ agonist significantly enhanced the intracellular lipid accumulation and CD36 expression that were induced by Mtb infection and down-regulated the lipid level in the supernatant of cell culture and cell proliferation rate, while PPARγ antagonist reversed the above effects. Conclusions:PPARγ played a role in lipid metabolism in Mtb-infected macrophages through affecting CD36 expression.

Five patients after prosthetic tricuspid valve, who received pacemaker implantation via coronary sinus during Oct, 2011 and Jul, 2014, were enrolled. Pacemakers were implanted via coronary vein in 5 patients without complications. The stimulation thresholds keep stable and symptoms (such as short breath and fatigue) were disappeared during the follow-up. For patients after tricuspid valve replacement, implantation of pacemaker via coronary sinus provides a safe and invasive approach and avoids opening the chest again.
Cardiac Surgical Procedures ; Coronary Sinus ; Heart Valve Prosthesis Implantation ; Humans ; Pacemaker, Artificial ; Tricuspid Valve

Intracranial atherosclerotic disease (ICAD) of a major intracranial artery, including middle cerebral artery (MCA), basilar artery, is the most common causes of stroke and is associated with a high risk of recurrent stroke in China. The difficulty to treatment these high-risk disease is to identify high-risk stroke subgroups and to develop more effective treatments (aggressive medical therapy/endovascular therapy). With the benefits, including non-invasive, in vivo, and no-ionizing radiation, 3.0 Tesla high-resolution magnetic resonance imaging (HR MRI) could be used to stratify high-risk patients, monitor progression of disease, and evaluate clinical efficacy, based on MCA wall structure and plaque characteristic. HR MRI has the latency of predicting high-risk patients benefit from endovascular therapy, having a broad application prospect during psot-SAMMPRIS era. The current research on MCA stenosis using HR MRI focuses on methodology, diagnosis and differential diagnosis, etiology, and lacks of clinical efficiency evaluation and prognostic analysis of ICAD treatment, especially lacks the research on in-stent restenosis, which needs further investigation.

Intracranial atherosclerotic disease (ICAD) of a major intracranial artery, including middle cerebral artery (MCA), basilar artery, is the most common causes of stroke and is associated with a high risk of recurrent stroke in China. The difficulty to treatment these high-risk disease is to identify high-risk stroke subgroups and to develop more effective treatments (aggressive medical therapy/endovascular therapy). With the benefits, including non-invasive, in vivo, and no-ionizing radiation, 3.0 Tesla high-resolution magnetic resonance imaging (HR MRI) could be used to stratify high-risk patients, monitor progression of disease, and evaluate clinical efficacy, based on MCA wall structure and plaque characteristic. HR MRI has the latency of predicting high-risk patients benefit from endovascular therapy, having a broad application prospect during psot-SAMMPRIS era. The current research on MCA stenosis using HR MRI focuses on methodology, diagnosis and differential diagnosis, etiology, and lacks of clinical efficiency evaluation and prognostic analysis of ICAD treatment, especially lacks the research on in-stent restenosis, which needs further investigation.

OBJECTIVETo assess the reproducibility of 3.0 T high-resolution magnetic resonance imaging (HR MRI) for evaluation of atherosclerotic stenosis in the middle cerebral artery (MCA).

METHODSFrom February, 2011 to December, 2013, 66 consecutive patients with MCA-M1 atherosclerotic stenosis (50%-99%) confirmed by digital subtractive angiography (DSA) received examinations with 3.0 T HR MRI for measurement of the vessel area (VA) and lumen area (LA) at the maximum narrow site (VA(narrow) and LA(narrow)) and the reference site (VA(reference) and LA(reference)) as well as the plaque distribution (ventral, dorsal, superior, and inferior). Two independent readers reviewed all the images and one reader reevaluated these images 4 weeks later. The inter- and intra-observer reproducibility was evaluated using the intraclass correlation coefficient (ICC).

RESULTSThe measurements of VA(narrow), VA(reference), and LA(reference) using HR MRI showed excellent inter- (ICC=0.801, 0.843, and 0.808, respectively) and intra-observer reproducibility (ICC=0.811, 0.916, and 0.958, respectively), but the measurement of LA(narrow) had only moderate inter- and intra-observer reproducibility (ICC=0.584 and 0.625, respectively). For plaque distribution analysis (ventral, dorsal, superior, and inferior plaques), HR MRI also showed excellent inter- (ICC=0.856, 0.836, 0.791, and 0.905, respectively) and intra-observer reproducibility (ICC=0.876, 0.827, 0.825, and 0.950, respectively).

CONCLUSIONHR MRI shows good inter- and intra-observer reproducibility in identifying MCA-M1 atherosclerotic plaque distribution and vessel and lumen measurements, but its reliability for lumen area measurement at the maximum narrowing site needs to be improved.