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 trend in incidence, causative drugs, clinical types and treatment of drug eruption. Methods Clinical data were collected from 922 inpatients with drug eruption in Huashan Hospital, Fudan University from January 2009 to December 2013, and analyzed retrospectively. Results From 2009 to 2013, the percentage of inpatients with drug eruption among all inpatients in the Department of Dermatology in a given year varied from 9.45% to 10.01%, and the percentage of inpatients with severe drug eruption among inpatients with drug eruption from 17.45% to 28.24%. Of the 922 cases, 371 (40.2%)were caused by single drugs, and 551 (59.8%)by multiple drugs. Among the 371 cases of drug eruption caused by single drugs, the top five causative drugs were traditional Chinese medicine(72 cases), cephalosporins(38 cases), amoxicillin(27 cases), antipyretic analgesics(26 cases)and tetanus antitoxin (24 cases)in 278 cases of non-severe drug eruption, antiepileptic agents (33 cases), allopurinol (28 cases), antipyretic analgesics (7 cases), cephalosporins (6 cases)and traditional Chinese medicine (6 cases)in 93 cases of severe drug eruption. Of the 922 patients, 422 (45.8%)presented with maculopapular eruption, 259 (28.1%)with urticaria, 135(14.6%)with Stevens-Johnson syndrome, 49(5.3%)with toxic epidermal necrolysis, 33(3.6%)with drug reaction with eosinophilia and systemic symptoms (DRESS), and 7 (0.8%)with acute generalized exanthematous pustulosis (AGEP). A total of 791 (85.8%)patients with drug eruption received glucocorticoid treatment. The dose of glucocorticoids was(47.61 ± 12.07)mg prednisone equivalent per day in 550 patients with non-severe drug eruption, and (73.10 ± 18.23)mg prednisone equivalent per day in 221 patients with severe drug eruption. Totally, 110 (11.0%) patients with drug eruption were treated with combined intravenous immunoglobulin (IVIG)because of poor response to glucocorticoids alone. Of 224 patients with severe drug eruption, only 2 (0.9%)died. Conclusions Carbamazepine and allopurinol are the main causative drugs for severe drug eruption, while traditional Chinese medicine is the first causative drug for non-severe drug eruption. From 2009 to 2013, the annual mortality of severe drug eruption decreased considerably.

Objective To investigate the effect of stellate ganglion block combined with morphine sustainedrelease tablets for controlling pain of advanced lung cancer.Methods 90 cases with advanced lung cancer were randomly divided into the observation group and control group with 45 cases in each group.The control group was givenmorphine sustained-release tablets,the observation group received combination therapy with stellate ganglion block and morphine sustained-release tablets.The analgesic effects,quality of life scores and adverse reactions of two groups were compared.Results The effective rate of observation group was 93.33 ％ (42/45),which was significantly higher than that of control group 77.78％ (35/45),the difference was statistically significant (x2 =4.54,P ＜ 0.05).The morphine consumption of observation group was significantly less than that of control group(t =9.11,P ＜ 0.05),duration of analgesia was significantly longer than that of control group (t =6.72,P ＜ 0.05).Quality of life score in the observation group was significantly higher than that in the control group,the difference was statistically significant(Z =3.61,P ＜ 0.05).The incidence rate of adverse reactions was 17.78％ (8/45),which was significantly lower than that of the control group 46.67％ (21/45),the difference was statistically significant (x2 =4.54,P ＜ 0.05).Conclusion Stellate ganglion block combined with morphine sustained-release tablets for controlling pain of advanced lung cancer can significantly improve analgesia,reduce morphine consumption and adverse reactions,is worthy of clinical application.

Objective To explore the relationship between the HLA-G 14 bp insertion/deletion polymorphism and the infection of Epstein-Barr virus(EBV) for children.Methods The study genotyped HLA-G 14 bp insertion/deletion polymorphism of 102 infectious mononucleosis children and 165 normal controls by PCR-PAGE,detected the plasma sHLA-G level of 51 infectious mononucleosis children and 146 normal controls by ELISA.Results A significant difference was observed for the frequencies of the HLA-G 14 bp genotype between the two groups( x2 =6.742,P=0.034 ),and a significant difference was also observed for the 14 bp allele frequencies between the two groups( x2 =6.672,P=0.01 ).The plasma sHLA-G levels in the infectious mononucleosis children were dramatically higher than that in normal controls,and a significant difference was observed between the two groups( Z=-9.472,P＜0.01 ).Among the infectious mononucleosis children,levels of sHLA-G was find a significant difference between the three genotypes of HLA-G 14 bp insertion/deletion polymorphism( H=6.09,P =0.048 ),and the level of s HLA-G with 14 bp+/+ genotype was markedly lower than that of the two other genotypes (Z=-2.376,P=0.01 8).Conclusion There was a relationship between the HLA-G 14 bp insertion/deletion polymorphism and the susceptibility to the infectious mononucleosis for children.Children who carried the 14 bp-/- genotype or deleted the 14 bp allele may have a significantly increased risk of the infection of EBV.The plasma sHLA-G might be considered as an index for auxiliary diagnosis infectious mononucleosis.

Objective To study the incidence and clinical significance of the herniation pits of the femoral neck.Methods 600 cases(299 men,301 women,18～82 years)were collected.The incidence,radiologic finding and clinical significance of the herniation pit of the femoral neck were analysed.Results Of 1200 hip joints in 600 cases,there was 58 cases(64 sides)(5.3%)with herniation pits of the femoral neck,including 39 men(68.7%)and 19 women(31.3%),the lesions localized in the left joint in 25 eases(39.1%),in the right joint in 27 cases(42.2%)and in bilateral joints in 6 cases(18.7%).There were a obvious significant differences on both sexes,and no significant differences on age groups.On X-ray film,the lesions appeared as a round radiolucency with thin clear sclerotic rim.Conclusion The incidence of the herniation pit of the femoral neck is 5.3%,which has a typical X-ray feature,and may indicate the femoroacetabular impingement.

Objective To investigate the abnormality of parameters of thyroid function and incidence of autoimmune diseases in children with vitiligo.Methods A total of 363 children with vitiligo,including 198 males and 165 females were recruited into this study together with 93 normal human controls(55 males and 38 females).The serum levels of free tetraiodothyronine,free triiodothyronine,thyroid stimulating hormone,antithyroperoxidase antibody and thyroglobulin antibody were determined by chemiluminescent immunoassay in these subiects.Results The abnormality of parameters of thyroid function was observed in 43 out of 363(11.8%)patients affected by vitiligo and in 4 out of 93(4.3%)normal human controls;a significant difference was observed between the two groups (P＜0.05).Of the 43 patients wim abnormality of parameters of thyroid function,39 were diagnosed as vitiligo vulgaris,4 as segmental vitiligo.A significant increase Was observed in the incidence of abnormality of parameters of thyroid function in patients with vitiligo vulgaris compared with those with segmental vitiligo(P＜0.05).Conclusion There is an increase in tbe abnormality of parameters of thyroid function in children with vitiligo.

Objective To analyze the diagnosis and therapy of pemphigus and pemphigoid.Methods Clinical data were retrospectively analysed for 648 patients with pemphigus or pemphigoid admitted in Huashan Hospital,Fudan University from 2003 to 2007.Results The mean age of onset of pemphigus was younger than that of pemphigoid(47.0±16.9 years vs 65.1±13.9 years.P＜0.001).Of 175 patients with pemphigus confirmed by direct immunoflorescence(DIF),100%were positive for IgG,and 92.0%for C3,while out of 223 patients with pemphigoid,51.12%were positive for IgG,99.1%for C3.For pemphigus,the consistency reached 68.8%among clinical diagnosis.pathological diagnosis and immunological(DIF)diagnosis,80.7%between pathological and immunological diagnosis.In the case of pemphigoid,the consistency was 62.8%among clinical,pathological and immunological diagnosis,and 78.1%between pathological and immunological diagnosis.Corticosteroids were primary treatment strategy for both pemphigus and pemphigoid.and prednisone of 0.5-1.5 mg per kilogram bodyweight per day could control the condition of most patients.Conclusions DIF could be used as an important diagnostic means for patients with pemphigus or pemphigoid which can not be confirmed by clinical manifestation or pathology.In primary hospitals,IgG and C3 are recommended for DIF testing when pemphigus is suspected,and C3 is recommended for suspected pemphigoid.