Objective:To investigate effect and mechanism of hydroxysafflor yellow A(HSYA)activating neuronal autophagy on cerebral ischemia-reperfusion injury through a combination of in vitro and in vivo experiments.Methods:SD rat MCAO/R model was established by improved suture method.Rats were randomly divided into sham surgery(Sham)group,MCAO/R group and MCAO/R+HSYA group,following indicators were detected to determine extent of cerebral ischemia-reperfusion nerve damage:Z-Longa neu-rological function score was detected,TTC staining to measure cerebral infarction area,and TUNEL staining to measure cell apopto-sis;Western blot was used to detect protein expressions of autophagy related markers LC3,Beclin1,P62 and SIRT1 in rat brain tis-sue;immunofluorescence staining was used to observe expression of LC3 co-localization with neurons.OGD/R injury model of SH-SY5Y cells was established and randomly divided into Normal group,OGD/R group,OGD/R+HSYA group,OGD/R+SIRT1 inhibitor(EX-527)group and OGD/R+EX-527+HSYA group.Western blot was used to detect protein expressions of LC3,Beclin1,P62 and SIRT1.Results:Compared with Sham group,model group rats showed impaired neurological function,significantly increased neu-robehavioral scores,widespread cerebral infarction,significantly increased neuronal cell apoptosis,significantly increased autophagy related protein Beclin1 expression and LC3-Ⅱ/LC3-Ⅰ,significantly decreased P62 expression,significantly increased LC3/NeuN co-stained cells,and decreased SIRT1 expression;compared with model group,HSYA intervention group showed a significant decrease in neurological functional scores,a significant reduction in cerebral infarction area,a significant decrease in neuronal cell apoptosis,a further increase in Beclin1 expression and LC3-Ⅱ/LC3-Ⅰ,a further decrease in P62 expression,number of LC3/NeuN and P62/NeuN co-stained cells also increased,and SIRT1 expression significantly increased.Expression trends of Beclin1,LC3-Ⅱ/LC3-Ⅰ,P62 and SIRT1 of cells between normal group,model group and HSYA intervention group were same as animal experiment;compared with model group,expressions of SIRT1,Beclin1 and LC3-Ⅱ/LC3-Ⅰ in OGD/R+EX-527 group were significantly reduced,while expression of P62 was significantly increased;compared with OGD/R+EX-527 group,there was no significant change in SIRT1 expression in OGD/R+EX-527+HSYA group,LC3-Ⅱ/LC3-Ⅰ and Beclin1 expression were significantly increased,and P62 expres-sion was significantly decreased.Conclusion:HSYA can significantly improve neurological deficits in rats after cerebral ischemia-reperfusion,reduce cerebral infarction area,and decrease neuronal cell apoptosis rate,whose neuroprotective effect may be related to its activation of SIRT1,which significantly enhances neuronal autophagy.

Objective:To explore effects and mechanism of Hydroxylsafflower Yellow A(HSYA)on expression of NLRP3 in glial cells after cerebral ischemic injury.Methods:A middle cerebral artery occlusion/reperfusion(MCAO/R)model was established in male SD rats.After successfully modeling for 24 h,Longa scoring and corner test were used to evaluate degree of neurological dys-function.Western blot and immunofluorescence were used to detect expressions of JAK2/STAT3 molecules and NLRP3,ELISA was used to measure IL-1β,IL-6 and TNF-α levels.A glucose-oxygen deprivation/reperfusion(OGD/R)model was established in microg-lia,and JAK2 and STAT3 inhibitor AG490 was used to further verify action of HSYA on NLRP3.Results:Compared with sham group,neurological dysfunction aggravated in MCAO/R group(P<0.01),HSYA treatment improved neurological function(P<0.01).Expres-sions of p-JAK2,p-STAT3 and NLRP3 in MCAO/R group were higher than those in the sham group(P<0.01);and HSYA treatment reduced expressions of p-JAK2,p-STAT3 and NLRP3(P<0.01).Levels of inflammatory factors IL-1β,IL-6 and TNF-α were higher in MCAO/R group than sham group(P<0.01),and HSYA inhibited expressions of IL-1β,IL-6 and TNF-α(P<0.01 or P<0.05).In vi-tro experiments showed expressions of p-JAK2,p-STAT3 and NLRP3 in OGD/R group were significantly higher than normal control group(P<0.01),after adding AG490,phosphorylation of JAK2 and STAT3 decreased,NLRP3 expression was inhibited(P<0.01).Inflammatory cytokines IL-1β,IL-6 and TNF-α levels were higher in OGD/R group than normal control group(P<0.01),and HSYA inhibited expressions of IL-1β,IL-6 and TNF-α(P<0.01 or P<0.05).Conclusion:HSYA alleviates brain damage,probably by regu-lating JAK2/STAT3 signaling pathway and inhibiting NLRP3 expression in microglia after cerebral ischemia and hypoxia.

Objective:To explore effects and mechanism of Hydroxylsafflower Yellow A(HSYA)on expression of NLRP3 in glial cells after cerebral ischemic injury.Methods:A middle cerebral artery occlusion/reperfusion(MCAO/R)model was established in male SD rats.After successfully modeling for 24 h,Longa scoring and corner test were used to evaluate degree of neurological dys-function.Western blot and immunofluorescence were used to detect expressions of JAK2/STAT3 molecules and NLRP3,ELISA was used to measure IL-1β,IL-6 and TNF-α levels.A glucose-oxygen deprivation/reperfusion(OGD/R)model was established in microg-lia,and JAK2 and STAT3 inhibitor AG490 was used to further verify action of HSYA on NLRP3.Results:Compared with sham group,neurological dysfunction aggravated in MCAO/R group(P<0.01),HSYA treatment improved neurological function(P<0.01).Expres-sions of p-JAK2,p-STAT3 and NLRP3 in MCAO/R group were higher than those in the sham group(P<0.01);and HSYA treatment reduced expressions of p-JAK2,p-STAT3 and NLRP3(P<0.01).Levels of inflammatory factors IL-1β,IL-6 and TNF-α were higher in MCAO/R group than sham group(P<0.01),and HSYA inhibited expressions of IL-1β,IL-6 and TNF-α(P<0.01 or P<0.05).In vi-tro experiments showed expressions of p-JAK2,p-STAT3 and NLRP3 in OGD/R group were significantly higher than normal control group(P<0.01),after adding AG490,phosphorylation of JAK2 and STAT3 decreased,NLRP3 expression was inhibited(P<0.01).Inflammatory cytokines IL-1β,IL-6 and TNF-α levels were higher in OGD/R group than normal control group(P<0.01),and HSYA inhibited expressions of IL-1β,IL-6 and TNF-α(P<0.01 or P<0.05).Conclusion:HSYA alleviates brain damage,probably by regu-lating JAK2/STAT3 signaling pathway and inhibiting NLRP3 expression in microglia after cerebral ischemia and hypoxia.

Objective:To investigate effect and mechanism of hydroxysafflor yellow A(HSYA)activating neuronal autophagy on cerebral ischemia-reperfusion injury through a combination of in vitro and in vivo experiments.Methods:SD rat MCAO/R model was established by improved suture method.Rats were randomly divided into sham surgery(Sham)group,MCAO/R group and MCAO/R+HSYA group,following indicators were detected to determine extent of cerebral ischemia-reperfusion nerve damage:Z-Longa neu-rological function score was detected,TTC staining to measure cerebral infarction area,and TUNEL staining to measure cell apopto-sis;Western blot was used to detect protein expressions of autophagy related markers LC3,Beclin1,P62 and SIRT1 in rat brain tis-sue;immunofluorescence staining was used to observe expression of LC3 co-localization with neurons.OGD/R injury model of SH-SY5Y cells was established and randomly divided into Normal group,OGD/R group,OGD/R+HSYA group,OGD/R+SIRT1 inhibitor(EX-527)group and OGD/R+EX-527+HSYA group.Western blot was used to detect protein expressions of LC3,Beclin1,P62 and SIRT1.Results:Compared with Sham group,model group rats showed impaired neurological function,significantly increased neu-robehavioral scores,widespread cerebral infarction,significantly increased neuronal cell apoptosis,significantly increased autophagy related protein Beclin1 expression and LC3-Ⅱ/LC3-Ⅰ,significantly decreased P62 expression,significantly increased LC3/NeuN co-stained cells,and decreased SIRT1 expression;compared with model group,HSYA intervention group showed a significant decrease in neurological functional scores,a significant reduction in cerebral infarction area,a significant decrease in neuronal cell apoptosis,a further increase in Beclin1 expression and LC3-Ⅱ/LC3-Ⅰ,a further decrease in P62 expression,number of LC3/NeuN and P62/NeuN co-stained cells also increased,and SIRT1 expression significantly increased.Expression trends of Beclin1,LC3-Ⅱ/LC3-Ⅰ,P62 and SIRT1 of cells between normal group,model group and HSYA intervention group were same as animal experiment;compared with model group,expressions of SIRT1,Beclin1 and LC3-Ⅱ/LC3-Ⅰ in OGD/R+EX-527 group were significantly reduced,while expression of P62 was significantly increased;compared with OGD/R+EX-527 group,there was no significant change in SIRT1 expression in OGD/R+EX-527+HSYA group,LC3-Ⅱ/LC3-Ⅰ and Beclin1 expression were significantly increased,and P62 expres-sion was significantly decreased.Conclusion:HSYA can significantly improve neurological deficits in rats after cerebral ischemia-reperfusion,reduce cerebral infarction area,and decrease neuronal cell apoptosis rate,whose neuroprotective effect may be related to its activation of SIRT1,which significantly enhances neuronal autophagy.

OBJECTIVE：To compare the c ontents of 6 kinds of ester alkaloids in raw aconite and 5 kinds of salt-processed products，and to provide reference for the optimization of salt-processing technology. METHODS ：Processed with bittern ， magnesium chloride ，sodium chloride ，calcium chloride and potassium chloride as excipients ，and HPLC method were adopted to determine the contents of 6 kinds of ester alkaloids in raw aconite and 5 kinds of salt-processed products. The effect/toxicity ratio and toxicity component index were used to evaluate the effect and toxicity of raw aconite and 5 kinds of salt-processed products. RESULTS： The linear range of benzoylneoaconitine ， benzoylhypoconitine， benzoylaconitine， neoaconitine， aconitine and hypoconitine were 2.440-24.40，2.240-22.40，2.020-20.20，2.780-27.80，2.240-22.40，2.240-22.40 μg/mL（r≥0.999 0）. RSDs of precision，stability and repeatability tests were all less than 2％. The recovery rates were 102.13％-104.87％（RSD＝0.98％，n＝ 6），100.00％-105.00％（RSD＝2.02％，n＝6），95.00％-99.29％（RSD＝1.77％，n＝6），100.41％-104.58％（RSD＝1.78％，n＝ 6），98.87％-99.90％（RSD＝0.41％，n＝6），100.20％-104.00％（RSD＝1.55，n＝6），respectively. The contents of total alkaloids in order from the largest to the smallest was as follows ：raw aconite ＞processed products of sodium chloride ＞processed products of bittern ＞processed products of potassium chloride ＞processed products of calcium chloride ＞processed products of magnesium chloride. The order of effect/toxicity ratio was processed products of potassium chloride ＞processed products of magnesium chloride＞processed products of sodium chloride ＞processed products of calcium chloride ＞processed products of bittern ＞raw aconite. The order of toxicity component index was raw aconite ＞processed products of sodium chloride ＞processed products of bittern＞processed products of calcium chloride ＞processed products of potassium chloride ＞processed products of magnesium chloride. CONCLUSIONS ：Using 5 kinds of salt as excipients ，the proce ssing technology has different degrees of “efficacy enhancing and toxicity reducing ”effect. Among them ，magne- sium chloride ，potassium chloride and calcium chloride are better for processing and can be used for processing aconite.

Objective To investigate the effects of ginkgo biloba extract on the early postoperative cognitive function and the serum neuron specific enolase (NSE),S100β,tumor necrosis factor-α (TNF-α),interleukin (IL)-6,superoxide dismutase (SOD) and malondialdehyde (MDA) in patients with meningioma resection.Methods 60 patients with meningioma scheduled for elective intracranial tumor resection were enrolled and randomly divided into two groups (n =30 each):ginkgo biloba extract group (group G) and control group (group C).Ginkgo biloba extract was infused intravenously 30 min before anesthesia induction in group G,while group C received the equal volume of normal saline.Venous blood sample were taken at three time points:30 min before induction (T1),extubation (T2) and 24 h after operation (T3)for determination of serum concentration of NSE,S100β,TNF-α,IL-6,SOD and MDA.Cognitive function was evaluated at 1 d before and 3 d after surgery using Mini-mental State Examination (MMSE),Wechsler Adult Intelligence Scale (WAIS) and Wechsler Memory Scale (WMS).Results Compared with T1,the concentration of NSE,S100β,TNF-α,IL-6 and MDA in serum were significantly increased at T2 and T3 in the two groups,while SOD was decreased (P ＜ 0.05).Compared with group C,the concentration of NSE,S100β,TNF-α,IL-6 and MDA were significantly decreased at T2 and T3 in group G,while SOD was increased (P ＜ 0.05).Compared with 1 d before surgery,the scores of MMSE,digit accumulation,digit breadth (forward),vocabulary association and digit symbol replacement were significantly decreased and tracing connection time was significantly increased on the 3rd day after surgery in the two groups.The scores of digital breadth (reverse) and visual regeneration were significantly decreased in group C (P ＜ 0.05).Compared with group C,the scores of digit accumulation,digit breadth (forward and reverse) and visual regeneration were significantly increased on the 3rd day after surgery in group G (P ＜ 0.05).The incidence of postoperative cognitive dysfunction (POCD) at 3 d after surgery in group G was significantly lower than group C (P ＜ 0.05).Conclusions The pretreatment of ginkgo biloba extract can decrease the incidence of early POCD in patients with meningioma resection,and its mechanism may be related to the reduction of inflammatory reaction and antioxidation.

Objective This study aimed to assess the diagnostic value of anti-mutated citrullinated vimentin (MCV) antibodies in rheumatoid arthritis (RA) and its correlation with disease progression, extra-articular manifestations and overlap syndrome. Methods Retrospective Studies. Clinical data of 837 patients in PekingUnionMedicalCollegeHospitalfrom June to August 2017 were collected, including the result of anti-MCV, anti-cyclic citrullinated peptide (anti-CCP) antibodies, rheumatoid factor (RF), erythrocyte sedimentation rate (ESR) and High-sensitivity-C-reactive protein (CRP). According to the 1987 American College of Rheumatology classification criteria for rheumatoid arthritis, there were 323 patients diagnosed with RA, including 59 males and 264 females with the average age of 51 years. According to whether the RA patients have overlap syndrome with other autoimmune disease (AID) or have extra-articular manifestations, 258 cases were categorized into RA group, including 47 males and 211 females with the average age of 50 years; 14 cases were categorized into the group of overlap syndrome, including 1 male and 13 females with the average age of 36 years;51 cases were categorized into the group of extra-articular manifestations, including 11 males and 40 females with the average age of 59 years.According to 2010 rheumatoid arthritis classification criteria for destruction in joints, the radiographic changes were divided into 4 stages. There were 203 casesenrolled in our study, 88 caseswere fitted into early stage group (stage I)including 21 males and 67 females with the average age of 48 years; 115 caseswere fitted into progressive stage group, which compromisedstageⅡ (interim stage), stage Ⅲ (severe stage) and stage Ⅳ(final stage) cases, including 19 males and 96 females with the average age of 53 years. Mann-Whitney U test, x2 test, Receiver operating characteristic (ROC) curves and Spearmancorrelation coefficientwere used in Statistical analysis. Results Ⅰ Amongdiagnosed RA patients, 199 (61.6％) cases were positive for anti-MCV, anti-CCP and RFsimultaneously, 42 (13％) cases were positive for anti-MCV, which was higher than anti-CCP positive (1 cases, 0.3％) or RF positive (7cases, 2.2％). The difference was statistically significant(P<0.001, P<0.001). ⅡROC was calculated and MCV=35.95 U/ml was used as best-fit cut-off value. The AUC for anti-MCV was 0.867, while the sensitivity was 80.5％and specificity was 80.9％.ⅢThe detection levels of anti-MCV (682.8 (106.4-1000.0)), anti-CCP (407 (4.0-1536.0)) and RF (82.8 (21.1-244.9)) in the group of progressive stage were higher than those in the group of early stage (114.5 (28.5-1000.0), 62.5 (5.0-1020.7), 50.1 (6.7-127.1)), which showed a significant difference(P<0.05, P<0.05, P<0.05). The anti-MCV, anti-CCP and RF were positively related to the degree of joint destruction (r=0.229, P<0.05;r=0.187, P<0.05;r=0.167, P<0.05);anti-MCV and anti-CCP were positively related to extra-articular manifestation (r=0.152, P<0.05;r=0.136, P<0.05). Conclusion Anti-MCV antibodies are more sensitive in patients with RA, and have complementary diagnostic value for anti-CCP and RF-negative patients; high levels of anti-MCV and anti-CCP in RA patients are associated with RA progression and extra-articular involvement.

Objective To observe the levels of interferon gamma-induced protein 10(IP-10)in children with acute viral and bacterial infection.Methods There were three groups:acute viral infection group(51 ca-ses),acute bacterial infection group(52 cases),and healthy control group(51 cases).Serum IP-10 levels were detected by enzyme linked immunosorbent assay(ELISA),and serum C reactive protein(CRP)were deter-mined with BNⅡ automatic protein analyzer.Results The level of serum IP-10 and CRP were significantly different in different groups(P<0.05).Compared with control group,the level of IP-10 and CRP were higher in acute viral infection group or in acute bacterial infection group(P<0.05).The level of IP-10 was higher in acute viral infection group than that in acute bacterial infection group(P>0.05).The level of CRP was higher in bacterial infection group than that in acute viral infection group(P<0.05).The IP-10 levels in viral Infec-tion patients were not correlated with the CRP levels.The area under curve(AUC),sensitivity and specificity of IP-10 and CRP were 0.688 and 0.873,35.0% and 79.6%,96.1% and 98.0%.When cut-off value of predic-tive probability was 0.713,sensitivity and specificity were increased to 82.5% and 100.0%.Conclusion Ser-um IP-10,CRP and predictive probability are valuable in the diagnosis of acute pathogen infection in children.

Objective To compare eye expression recognition in stable outpatients with schizophrenia with that in normal controls and to explore the relationships between eye expression recognition and social functioning.Methods 107 schizophrenic outpatients and 66 normal controls matched in age,sex and years of education were assessed with Eye Basic Emotion Discrimination Task(EBEDT) and Eye Complex Emotion Discrimination Task(ECEDT).The patients were also assessed with Social Disability Screening Schedule (SDSS).Results The correct numbers were significantly lower for patients to identify basic emotions of eye expressions(13.2±3.8 vs16.0±2.6,P<0.01) and complex emotions of eye expressions(17.9±4.3 vs 20.6±3.5,P<0.01)than those for controls respectively;the correct numbers to identify anger(3.1±1.0 vs.2.1±1.2,P<0.01),fear(1.8±1.0 vs 1.3±1.0,P<0.01) and disgust(1.8±1.1 vs 1.4±1.2,P<0.05)for controls were higher than those for patients significantly.The correct numbers to identify total basic emotions(r=-0.335,P<0.05)and total complex emotions (r=-0.374,P<0.05)in eye expressions showed negatively correlated with the total scores of SDSS in the patients after controlling age and total score of PANSS.Conclusions The ability to recognize basic and complicated emotions in eye expressions in the outpatients with schizophrenia is lower than that in the controls. It shows positively correlated with social functioning moderately in the patients.

Objective To study the effect of VacA on the secretion of THP-1 macrophages as an individual virulence determinant, and the effect of NF-kB on the secretion of THP-1 macrophages. Methods The recombinant plasmid pDsRed-Monomer-Cl/vacA was transfected into macrophages. The cytokine con-tent of TNF-α or IL-1β in the culture medium was tested quantitatively with ELISA kit, respectively. The content of NO or ROS in the culture medium was tested with Griess reagent or DCFH-DA fluorescent probe. The apoptosis rate of macrophages was tested by flow cytometry. The effect of PDTC, an inhibitor of NF-kB, on the secretion and apoptosis of macrophages transfected with the recombinant plasmids, was also studied. The activity of NF-kB was examined in THP-1 cells by electrophoretic mobility gel shift assay(EMSA). Re-suits At 6 h after transfection, the level of TNF-α and IL-1 β in macrophages transfected with the recombi-nant plasmids was significantly higher than that of the control group (P <0.05). At 6 h or 12 h after trans-fection, the level of NO and ROS in macrophages transfected with the recombinant plasmids was significantly higher than that of the control group (P <0.05). At 16 h after transfection, the apoptosis rate of macropha-ges transfected with the recombinant plasmids was significantly higher than that of the control group (P < 0.05). PDTC decreased the production of TNF-α, IL-1 β, NO, ROS and apoptosis rate induced by VacA. VacA was found to trigger NF-kB activation. Conclusion The over-expression of VacA fusion protein can up-regulate secretion and apoptosis of macrophages. Activation of NF-kB is probably involved in the produc-tion of TNF-α, IL-1β, NO, ROS and apoptosis induced by VacA.